INNOVATIONS IN POSTHARVEST TECHNOLOGY SERIES

Postharvest Ripening
Physiology of Crops

Edited by
Sunil Pareek
Postharvest Ripening
Physiology of Crops
Series Editor
Sunil Pareek
Department of Agriculture and Environmental Sciences
National Institute of Food Technology Entrepreneurship and Management
Kundli, Sonepat, Haryana, India

Postharvest Ripening Physiology of Crops (2016)

Edited by Sunil Pareek
Postharvest Ripening
Physiology of Crops
Edited by
Sunil Pareek

Boca Raton London New York

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 Contents

DEDICATION XXI
SERIES PREFACE XXVII
FOREWORD XXIX
PREFACE XXXI
ACKNOWLEDGMENTS XXXIII
EDITOR XXXV
CONTRIBUTORS XXXVII

1 Ripening Physiology: An Overview 1

SUNIL PAREEK
Abstract 2
1.1 Introduction 2
1.2 Climacteric Phenomenon 4
1.3 Physicochemical and Metabolic Changes 12
1.3.1 Color Changes 13
1.3.2 Sugar Changes 14
1.3.3 Organic Acid Changes 16
1.3.4 Flavor and Aroma Changes 18
1.3.5 Cell Wall and Textural Changes 22
1.3.6 Physiological Changes 28
1.4 Conclusions and Future Perspectives 33
References 33

2 Postharvest Physiology of Fruits and Vegetables 49

PETER M.A. TOIVONEN
Abstract 50
2.1 Introduction 51

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2.2 Classification of Fruits and Vegetables Based on

Physiological Characteristics 51
2.2.1 Ethylene Biology 51
2.2.2 Respiratory Characteristics: Climacteric and
Nonclimacteric 56
2.2.3 Developmental or Maturity Stage at Harvest 56
2.2.4 Tolerance to Low Temperatures 58
2.3 Factors Affecting Physiology of Fruits and Vegetables in
Postharvest Systems 60
2.3.1 Temperature 60
2.3.2 Humidity 62
2.3.3 Atmospheric Modification 64
2.3.4 Abiotic Stresses 65
2.4 Physiological Changes Occurring during Postharvest
Handling or Storage 66
2.4.1 Depletion of Respiratory Substrate 66
2.4.2 Hormonal Effects 68
2.4.3 Membrane Alterations 71
2.5 Conclusions and Future Perspectives 71
References 72

3 Postharvest Quality of Ornamental Plants 81

FERNANDO L. FINGER, TANIA P. SILVA, FERNANDA
F. ARAUJO, AND JOSE G. BARBOSA
Abstract 82
3.1 Introduction 82
3.2 Quality Attributes in Ornamental Plants 83
3.3 Influence of Water Relations on Ornamental Longevity 85
3.4 Action of Ethylene on Ornamental Plant Quality 88
3.4.1 Inhibition of Ethylene Synthesis 91
3.4.2 Inhibition of Ethylene Action 93
3.4.3 Ethylene Absorbers 95
3.5 Role of Abscisic Acid, Gibberellins, and Cytokinins 96
3.6 Role of Calcium on Flower Senescence 97
3.7 Respiration 98

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3.8 Temperature 99
3.9 Handling of Cut Flowers 102
3.10 Potted Plants 103
3.11 Conclusions and Future Perspectives 104
Acknowledgment 105
References 105

4 Physiology and Molecular Biology of

Flower Senescence 109
KENICHI SHIBUYA AND KAZUO ICHIMURA
Abstract 110
4.1 Introduction 110
4.2 Ethylene and Senescence of Cut Flowers 111
4.2.1 Ethylene Response of Cut Flowers 111
4.2.2 Types of Senescence in Cut Flowers with High
Ethylene Sensitivity 112
4.2.3 Ethylene in Petal-Wilting-Type Flowers 113
4.2.4 Ethylene in Petal-Abscission-Type Flowers 114
4.2.5 Ethylene Biosynthesis 114
4.2.6 Ethylene Signal Transduction 115
4.2.7 Acceleration of Flower Senescence by
Pollination 116
4.2.8 Acceleration of Flower Senescence by
Wounding 119
4.2.9 Effect of Temperature on Ethylene Production
and Perception 119
4.3 Plant Hormones Other than Ethylene in Flower
Senescence 120
4.3.1 Auxin 120
4.3.2 Gibberellin 120
4.3.3 Cytokinin 120
4.3.4 Abscisic Acid 121
4.3.5 Jasmonic Acid 122
4.4 Programmed Cell Death in Flower Senescence 122
4.4.1 Programmed Cell Death 122

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4.4.2 Gene Expression during PCD in Flowers 123

4.4.3 Autophagy in Petal Senescence 124
4.5 Conclusions and Future Perspectives 124
References 125

5 Respiratory Metabolism 139

MIKAL E. SALTVEIT
Abstract 140
5.1 Introduction 140
5.2 Why Measure Respiration? 141
5.3 Major Components of Respiration 142
5.3.1 Glycolysis 142
5.3.2 Pentose-Phosphate Shunt 142
5.3.3 Anaerobic Diversion 143
5.3.4 Tricarboxylic Acid Cycle 143
5.3.5 Electron Transport (Chemiosmotic
Phosphorylation) 145
5.4 Measurement of Respiration 146
5.4.1 Loss of Substrate, Heat Production, and Water 146
5.4.2 Consumption of Oxygen and Production of
Carbon Dioxide 147
5.4.2.1 Static System 147
5.4.2.2 Flow-Through or Dynamic System 149
5.5 Sampling and Analyzing 151
5.6 Instruments and Techniques 152
5.7 Pre- and Postharvest Factors Affecting Respiration 152
5.7.1 Temperature Effects 152
5.7.2 Respiratory Quotient 154
5.7.3 Physical Stress 154
5.7.4 Internal Factors 155
5.7.4.1 Genotype 155
5.7.4.2 Type of Plant Part 155
5.7.4.3 Respiratory Climacteric 155
5.8 Conclusions and Future Perspectives 156
References 156

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6 Stomata and Postharvest Physiology 157

UULKE VAN MEETEREN AND SASAN ALINIAEIFARD
Abstract 158
6.1 Introduction 158
6.2 Stomata 159
6.2.1 Role of Stomata in Plants 159
6.2.2 Mechanism of Stomatal Closure and Opening 161
6.2.3 Signal Transduction Pathways in Guard Cells
for Stomatal Closure 164
6.3 Role of Stomata in the Postharvest Phase 166
6.3.1 Stomata in Relation to Vase Life of Cut Flowers 166
6.3.2 Stomata in Relation to Quality of Vegetables 169
6.3.3 Stomata in Relation to Quality of Fruits 172
6.4 Preharvest Conditions Leading to Postharvest Problems
via Stomata 174
6.4.1 Relative Humidity and Stomata Control 175
6.4.1.1 How Does Preharvest Low VPD
Affect Postharvest Stomata Control? 176
6.4.1.2 Induction of Stomata Morphological
Changes by Low VPD 179
6.4.2 Temperature 181
6.4.3 Light 182
6.5 Stomata and Tolerance to Postharvest Diseases and
Physiological Disorders 184
6.6 Postharvest Treatments and Stomata 186
6.7 Ethylene, Stomata, and Senescence 187
6.8 Conclusions and Future Perspectives 190
References 191

7 Water Loss from Harvested Horticultural

Commodities 217
MIKAL E. SALTVEIT
Abstract 218
7.1 Importance of Water Loss 219
7.2 Properties of Water 220

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7.3 Psychrometrics: Behavior of Water in Air 221

7.4 Transpiration: Diffusion of Water Vapor 224
7.5 Resistance to Diffusion of Water Vapor 226
7.6 Measurement of Transpiration 226
7.6.1 Weight Loss 227
7.6.2 Direct Measurement 227
7.6.3 Diffusion Porometer 227
7.7 Factors Affecting Water Loss 227
7.7.1 Commodity Factors 227
7.7.1.1 Surface-to-Volume Ratio 228
7.7.1.2 Routes of Water Loss 228
7.7.1.3 Anatomy of the Evaporating Surface 229
7.7.1.4 Physiological State of the
Commodity 230
7.7.1.5 Cultivar 230
7.7.1.6 Cultural Conditions 231
7.7.2 Environmental Factors 231
7.7.2.1 Humidity 231
7.7.2.2 Diffusion Shells and Air Velocity 231
7.7.2.3 Temperature 232
7.7.2.4 Atmospheric Pressure 232
7.8 Methods to Reduce Water Loss 232
7.8.1 Handling Techniques 232
7.8.2 Proper Refrigeration Design 232
7.8.3 Packaging 233
7.8.4 Waxing 234
7.8.5 Film Wraps 234
7.8.6 Curing 234
7.9 Conclusions and Future Perspectives 235
References 235

8 Lysophospholipids and Postharvest Quality

of Fruits, Vegetables, and Cut Flowers 237
DOMINGOS P.F. ALMEIDA
Abstract 238
8.1 Introduction 238

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8.2 Lipids as Signal Molecules in Plant Senescence and

Stress Response 239
8.2.1 Chemistry 239
8.2.2 Metabolism 240
8.3 Modulation of Postharvest Quality by Lysophospholipids 241
8.3.1 Effects on Color 242
8.3.2 Effects on Texture 242
8.3.3 Other Effects on Postharvest Quality-Related
Features 246
8.4 LPE Treatment Condition 247
8.5 Improvement of Postharvest Quality by
Lysophospholipids: Potential and Limitations 248
8.6 Conclusions and Future Perspectives 249
References 249

9 Fruit Skin Color and the Role of Pigments

during Fruit Ripening 255
EMRUL KAYESH, LINGFEI SHANGGUAN, AND
M. MOFAZAL HOSSAIN
Abstract 256
9.1 Introduction 256
9.2 Economics of Color Fruit 258
9.3 Carotenoid Pigments in Fruits 259
9.4 Anthocyanin Pigments in Fruits 260
9.5 Grape as a Model System for Pigmentation Studies
in Fruit Crops 270
9.6 Case Studies 278
9.6.1 Apple 278
9.6.2 Strawberry 279
9.6.3 Tomato 280
9.6.4 Litchi 281
9.6.5 Chinese Bayberry 282
9.6.6 ‘Hass’ Avocados 283
9.6.7 Pear 283
9.6.8 Cherry 284
9.6.9 Kiwifruit 284

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9.7 Pigments from Fruit to Human Health 284

9.8 Transgenic Plants Developed for Fruit Color 285
9.9 Conclusions and Future Perspectives 286
References 288

10 Molecular Regulation of Fruit Ripening 299

AJAY ARORA
Abstract 300
10.1 Introduction 300
10.2 Transcriptional Control of Fruit Ripening 301
10.3 Hormonal and Transcriptional Regulation during
Ripening 305
10.3.1 Ethylene 306
10.3.2 Auxins 308
10.3.3 Gibberellins 308
10.3.4 Abscisic Acid 308
10.4 Epigenetic Regulation of Fruit Development and
Ripening 309
10.5 Ripening Pathways and Associated Fruit Quality 312
10.5.1 Sugar Accumulation 313
10.5.2 Cell Walls and Fruit Shelf Life 313
10.5.3 Color, Flavor, and Nutrition in Fruits 314
10.5.3.1 Chloroplast-to-Chromoplast
Conversion 314
10.5.3.2 Flavor and Aroma Production 315
10.5.3.3 Control of Color and Texture
Changes 316
10.6 Human Nutrition and Fruits 317
10.6.1 Human Nutrition: Functional Genomics/
Systems Approach 320
10.7 Horticultural Crop Improvement 321
10.8 Genetic Manipulation of Ripening Regulatory
Genes 322
10.9 Conclusions and Future Perspectives 322
References 324

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11 Advances in Ethylene Signal Transduction in

Fruits and Vegetables 339
WILLIS O. OWINO AND JANE AMBUKO
Abstract 340
11.1 Introduction 340
11.2 Ethylene Physiology in Climacteric and Nonclimacteric
Fruits 341
11.3 Ethylene Signaling Components in Fruits 342
11.4 Analyses of Ethylene Signal Components in Other Fruit
Species 344
11.4.1 Climacteric Fruits 344
11.4.2 Nonclimacteric Fruits 348
11.5 Transcriptional Regulators of Fruit Ripening 349
11.6 Conclusions and Future Perspectives 352
References 353

12 Internal Atmosphere of Fruits: Role and

Significance in Ripening and Storability 359
VIJAY PAUL AND RAKESH PANDEY
Abstract 360
12.1 Introduction 361
12.2 Endogenous Volatiles in Fruits 363
12.3 Factors Affecting and Basis of Internal Atmosphere
of Harvested Fruit 363
12.4 Variability in the Internal Atmosphere of Fruits 366
12.5 Influence of the Internal Atmosphere on Ripening
and Related Aspects 367
12.5.1 Ripening 367
12.5.2 Flavor and Aroma 368
12.5.3 Fruit Decay 370
12.6 Ripening Behavior of Some Fruits under Attached
and Detached Conditions 371
12.6.1 Tomato 371
12.6.2 Melons 372
12.6.3 Sweet Pepper 372
12.6.4 Saskatoon 373

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12.7 Role of Some Gases and Endogenous Volatiles in

Fruit Ripening 373
12.7.1 Ethylene 373
12.7.1.1 Role of Ethylene in Ripening of
Climacteric Fruits 373
12.7.1.2 Role of Ethylene in Ripening of
Some Nonclimacteric Fruits 375
12.7.2 Oxygen and Carbon Dioxide 379
12.7.2.1 Low Oxygen 379
12.7.2.2 High Carbon Dioxide 382
12.7.2.3 Ratio of O2 to CO2 383
12.7.3 Ethanol and Acetaldehyde 384
12.7.4 Water Vapors and Water Status in Fruit 385
12.7.5 Salicylic Acid and Methyl Salicylate 387
12.7.6 Jasmonic Acid and Jasmonates 387
12.7.7 Nitric Oxide 388
12.8 Internal Atmosphere of Fruits: Practical Implication
in Ripening and Storability 388
12.9 Conclusions and Future Perspectives 390
References 391

13 Proteomics of Fruit Development and

Ripening 413
ALOYSIUS WONG, LUDIVINE THOMAS, CHRISTOPH GEHRING,
AND CLAUDIUS MARONDEDZE
Abstract 414
13.1 Introduction 414
13.2 Characteristics of Climacteric and Nonclimacteric
Fruit Ripening 416
13.3 Proteomic Tools and Approaches for Fruit Ripening
Assessment 417
13.3.1 Challenges in Proteomics Analysis 418
13.3.2 Protein Sample Preparation: Tissue Disruption,
Homogenization, and Solubilization 418
13.3.3 Protein Separation: Gel-Based and Gel-Free
Technologies 421

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13.4 Proteomes of Climacteric and Nonclimacteric Fruits 422

13.4.1 Overrepresented Functional Categories
during Development and Ripening 422
13.5 Pathway Analysis Using KEGG 426
13.5.1 Proteins Associated with Carbohydrate
Metabolism 426
13.5.2 Proteins Associated with Energy Metabolism 431
13.5.2.1 Proteins with a Role in Carbon
Fixation 431
13.5.2.2 Proteins Associated with the
Pentose Phosphate, Glycolysis,
and Pyruvate Metabolisms 432
13.5.2.3 Proteins Involved in the
TCA Cycle 436
13.5.3 Proteins Involved in Amino Acid Metabolism
and Ethylene Biosynthesis 436
13.5.4 Proteins Involved in Flavonoid Biosynthesis 437
13.6 Conclusions and Future Perspectives 438
References 439

14 Potato Tuber Dormancy and Postharvest

Sprout Control 449
JEFFREY C. SUTTLE, MICHAEL A. CAMPBELL, AND
NORA L. OLSEN
Abstract 450
14.1 Introduction 451
14.2 Dormancy: General Considerations 451
14.2.1 Developmental Aspects 452
14.3 Genetics of Tuber Dormancy 453
14.4 Pre- and Postharvest Environmental Effects 453
14.5 Physiological Regulation of Tuber Dormancy 454
14.5.1 Auxin 455
14.5.2 Carotenoid-Derived Hormones:
Abscisic Acid and Strigolactone 456
14.5.3 Cytokinins 458
14.5.4 Ethylene 459

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14.5.5 Gibberellins 460

14.5.6 Summary and Conclusions 460
14.6 Transcriptional Regulation during Dormancy 461
14.6.1 Initiation of Tuber Dormancy 461
14.6.2 Dormancy Termination 462
14.6.3 Epigenetic Control of Tuber Dormancy 463
14.7 Control of Sprouting in Storage 463
14.7.1 Introduction and Importance 463
14.7.2 Dormancy, Cultivar Selection, and
Storage Temperature 464
14.7.3 Chemical Control 464
14.8 Conclusions and Future Perspectives 467
References 467

15 Calcium Deficiency Disorders in Plants 477

SERGIO TONETTO DE FREITAS, CASSANDRO VIDAL TALAMINI
DO AMARANTE, AND ELIZABETH J. MITCHAM
Abstract 478
15.1 History of Ca2+ Deficiency Disorders 479
15.2 Role of Ca as an Essential Plant Macronutrient
2+ 479
15.3 Symptoms of Ca2+ Deficiency Disorders
in Fruit 481
15.3.1 Apple 481
15.3.2 Tomato 484
15.3.3 Watermelon 484
15.3.4 Pepper 484
15.4 Symptoms of Ca2+ Deficiency Disorders in
Leafy Vegetables 485
15.4.1 Lettuce 485
15.4.2 Cauliflower 485
15.4.3 Artichoke 486
15.4.4 Celery 487
15.5 Potential Mechanisms Regulating Ca2+ Deficiency
Disorders 488
15.5.1 Total Tissue Ca2+ Content 488

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15.5.2 Cellular Regulation of Ca2+ Partitioning

and Distribution 490
15.5.3 Other Nutrients 492
15.5.3.1 Nitrogen 492
15.5.3.2 Potassium and Magnesium 493
15.5.3.3 Boron 494
15.5.3.4 Phosphorus 494
15.5.4 Reactive Oxygen Species 495
15.5.5 Growth Regulators 495
15.5.5.1 Growth Regulators Affecting
Total Tissue Ca2+ Content 496
15.5.5.2 Growth Regulators Influencing
Cellular Ca2+ Distribution 498
15.5.5.3 Growth Regulator Effect on
Oxidative Metabolism 499
15.6 Possible Control Strategies 500
15.6.1 At the Tissue Level 500
15.6.2 At the Cellular Level 501
15.7 Final Considerations and Future Research
Needs 501
References 502

16 Fresh Fruit Aroma: An Integrative Overview

for a Complex Flavor Trait 513
ORIANNE GUDENSCHWAGER AND
BRUNO G. DEFILIPPI
Abstract 514
16.1 Introduction 515
16.2 Aroma Composition in Fruits 516
16.2.1 Apple 516
16.2.2 Melon 517
16.2.3 Strawberry 517
16.2.4 Tomato 518
16.2.5 Citrus 518
16.2.6 Grape 519

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16.2.7 Peach 519

16.2.8 Banana 520
16.3 Biosynthesis and Regulation of Aroma Volatiles in Fruit 520
16.3.1 Biosynthetic Pathways of Aroma Volatiles 520
16.3.1.1 Fatty Acid Metabolism 520
16.3.1.2 Amino Acid Metabolism 522
16.3.1.3 Ester Biosynthesis 522
16.3.1.4 Carbohydrate Metabolism 524
16.3.2 Aroma Modulation during Fruit Ripening 525
16.4 Influence of Pre- and Postharvest Factors on Fruit
Aroma 527
16.4.1 Preharvest Factors 527
16.4.1.1 Genotype 527
16.4.1.2 Growing Conditions 529
16.4.1.3 Fruit Maturity 531
16.4.2 Postharvest Technologies 532
16.4.2.1 Storage Temperature 532
16.4.2.2 Storage Atmosphere 534
16.4.2.3 Ethylene Control 536
16.4.2.4 Other Technologies 537
16.5 Conclusions and Future Perspectives 538
Acknowledgments 539
References 539

17 Flavor and Aroma Compounds of Some

Exotic Tropical Fruits and Berries: Biosynthetic
Pathways and Metabolism 553
OLA LASEKAN
Abstract 554
17.1 Introduction 555
17.2 Exotic Fruits and Their Flavor Profiles 556
17.2.1 Lychee (Litchi chinensis) 556
17.2.2 Rambutan (Nephelium lappaceum L.) 556
17.2.3 Yellow Passion (Passiflora edulis) 558
17.2.4 Durian Fruit (Durio zibethinus) 559

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17.2.5 Star Fruit or Carambola (Averrhoa carambola L.) 560

17.2.6 Mangosteen (Garcinia mangostana) 560
17.2.7 Snake Fruit (Salacca edulis Reinw) 560
17.2.8 Costa Rican Guava (Psidium
friedrichsthalium) 565
17.2.9 Pitanga Fruit (Eugenia uniflora L.) 566
17.2.10 Umbu-Caja Fruit (Spondias citherea) 566
17.2.11 Camu-Camu Fruit (Myrciaria dubia) 566
17.2.12 Cupuacu Fruit (Theobroma grandiflorum) 567
17.2.13 Araca-Boi Fruit (Eugenia stipitata) 567
17.2.14 Mangaba Fruit (Hancornia speciosa
Gomes)567
17.2.15 Garcinia Fruit (Garcinia dulcis Kurz) 568
17.2.16 Guabiju Fruit (Myrcianthes pungens Berg) 568
17.2.17 Guabiroba Fruit (Campomanesia xanthocarpa
Berg)568
17.2.18 Bacuri Fruit (Platonia sculenta) 568
17.2.19 Cashew Fruit (Anacardium occidentale L.) 569
17.2.20 Melon Fruit (Cucumis melo) 569
17.2.21 Jackfruit (Artocarpus heterophyllus Lam.) 570
17.2.22 Sapodilla Fruit (Achras sapota L.) 570
17.2.23 Genipap Fruit (Genipa americana) 571
17.2.24 Soursop Fruit (Annona muricata) 571
17.2.25 Acerola Fruit (Malphigia glabra L.) 572
17.2.26 Tamarind Fruit (Tamarindus indica L.) 572
17.2.27 Velvet Tamarind (Dialium guineense) 572
17.2.28  African Star Apple Fruit (Chrysophillum albidum) 573
17.3 Aroma Compounds’ Biosynthetic Pathways and
Metabolism 573
17.3.1 Fruit’s Volatile Ester Metabolism 573
17.3.2 Fruit’s Volatile Terpenoid Metabolism 575
17.3.3 Sulfur Volatile Compounds’ Biosynthetic
Pathway 577
17.4 Conclusions and Future Perspectives 579
References 579

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18 Impact of Postharvest Technologies on the

Flavor of Fresh Fruits and Vegetables 585
CHARLES F. FORNEY
Abstract 586
18.1 Introduction 587
18.2 Flavor of Fruits and Vegetables 588
18.2.1 Sensory Assessment 588
18.2.2 Chemical Flavor Constituents 589
18.3 Mechanism of Flavor Change 590
18.3.1 Metabolic Changes 591
18.3.2 Diffusional Changes 592
18.4 Impact of Postharvest Technologies 593
18.4.1 Ripening Manipulation 593
18.4.2 Temperature and Cold Storage 595
18.4.3 Controlled Atmosphere Storage 597
18.4.4 Packaging 599
18.4.5 Edible Coatings 601
18.4.6 Postharvest Treatments 601
18.4.6.1 Cutting 602
18.4.6.2 Heat Treatments 603
18.4.6.3 Irradiation 604
18.4.6.4 Ozone 605
18.4.6.5 Chemical Fumigation 606
18.5 Flavor Enhancement 607
18.6 Conclusions and Future Perspectives 607
References 608

xx
 Dedication

To the Late Professor Adel A. Kader

During his more than 35 years at the University of California, Davis (UC
Davis), Professor Adel Kader maintained an extremely active teaching,
research, and extension program in postharvest biology and technology of
horticultural crops. His prodigious output includes more than 230 technical
manuscripts, many book chapters, and numerous extension publications. He
was the most cited author by postharvest colleagues. Kader was the undis-
puted leader of a research team that has reached high levels of knowledge
in the different disciplines; he was a reference for researchers, enterprises,
and international institutes. His research efforts were initially focused on
improving the more obvious aspects of the postharvest quality of fruits and
vegetables. He worked with a large number of fruits and vegetables during
his career in his endeavor to elucidate the physiological and biochemical
bases for quality maintenance. His papers have been milestones for others
to follow. Over time, he became increasingly interested in the less obvious
characteristics of flavor and nutritional quality. He realized that an improve-
ment in human diets through the consumption of more fruits and vegetables
would occur only when the appearance quality of fruits and vegetables was
matched by an increase in their flavor and nutritive quality. He became a
vocal proponent for coupling the traditional studies for improved appear-
ance quality with studies of flavor and nutritional quality changes during
harvest, storage, transport, and marketing. All this because he recognized
the importance of bringing to the end user a product that not only looks
great but also tastes wonderful and is at its optimal nutritive value.

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D ed ic ati o n

Since beginning as a graduate student, Adel maintained a deep and

abiding interest in studying postharvest physiology and ensuring that fresh
fruits and vegetables are available to consumers in the best possible con-
dition. While a research assistant and as a technician, he was associated
with the late Dr. E.C. Maxie in pioneering studies on the effects of gamma
irradiation on the storability of fresh produce. His research was focused on
the responses of fruits to stress caused by O2 and CO2 during postharvest
handling. It involved the postharvest physiology of fruits, including mode
of action of oxygen and carbon dioxide on respiratory metabolism, ethylene
biosynthesis and action, and phenolic metabolism of fruits. Dr. Kader and
his coworkers developed new models and indices to predict fruit tolerance
for combinations of factors in controlled environments. They developed a
database for modified atmosphere packaging of fresh produce and iden-
tified optimum O2, CO2, and ethylene concentrations for storage of stone
fruits, Asian pear, kiwifruit, strawberry, and other commodities.
Professor Kader’s distinguishing characteristics included an amaz-
ing capacity to assimilate and organize information. He was universally rec-
ognized as among the most knowledgeable scientists in his field; however,
he was also widely known for freely sharing his knowledge and experience
with both the scientific community and the public. Intelligence, qualifica-
tion, organization, precision, and punctuality were the work tools that led
him to reach extraordinary scientific results. He also took the time to help
people and form lasting relationships. He was widely acknowledged for his
enthusiasm in teaching and in sharing his knowledge with others without
self-interests. At the age of 70, he organized many one-week short courses
and seminars with a few colleagues in Spain, Italy, Greece, India, Malaysia,
and UC Davis.
Few individuals, if any, can match the standards set by Professor
Kader during his tenure as a mentor. When it came to research integrity and
reaching for the highest standards possible, he practiced what he preached.
He demanded the highest performance from all colleagues, whether at UC
Davis or elsewhere. He took his responsibilities as a mentor very seriously.
One of Professor Kader’s most long-standing and sincerely held beliefs was
that students should be encouraged and given the means to attend scien-
tific meetings as a way of inspiring them in their developing careers. He
provided funding for countless students to travel to professional meetings
and advocated for allocating funds to support student travel, thus providing
the framework for the individual student to establish networking skills with
other scholars. Dr. Kader always had a gift handy in his bag: a T-shirt, a box
of chocolates, herb infusions, pens, a needle thermometer for fruit tempera-
ture, and sometimes the bag itself. Especially with young students, not only
did he dispense his knowledge through his lectures and seminars, but also
he spread “hard knowledge.” We all remember him recognizing the young-
est in the audience and presenting them with CDs, pen drives, or hard-copy

xxii
 D ed ic ati o n

books. During his years at UC Davis, he had a constant stream of interna-

tional visitors. He fostered an interactive environment for students and vis-
iting scientists, helping students to further develop networking skills and
expanding their vision beyond the boundaries of UC Davis.
Professor Kader’s in-depth knowledge of postharvest was a challenge
in itself to anyone discussing topics of mutual interest. He challenged not
only his students and visiting scientists, but also the larger worldwide post-
harvest community, to question research results and resolve the seemingly
unsolvable questions that inevitably arise from research. He instilled a
desire to learn in his students. The positive aspect of this challenging envi-
ronment was the intellectual growth that occurred, because he nurtured
this growth by positive feedback, thoughtful comments, and critiques.
Adel Kader was a constant and inspiring role model. He was always
organized (everyone was in awe of his perfectly organized desk and his
ability to instantly find a journal reference in his files), always prepared,
and always ready to listen to anyone or extend a helping hand. From the
perspective of postharvest biologists, Dr. Kader’s signature achievement
has to be the development of the UC Davis Postharvest Technology Center.
From a loose affiliation of postharvest extension specialists, who published
sporadic issues of a postharvest bulletin, he developed what is widely recog-
nized as the world’s best source for postharvest information and education.
His vision established the annual postharvest short course, which is now
in its 35th year. The impact of this course, with more than 2500 alumni,
including students, researchers, teachers, regulators, and postharvest
practitioners from around the world, is incalculable. To ensure the center’s
continued vitality, he also developed, and was a tireless advocate for, the UC
Davis Postharvest Program Endowment Fund.
Professor Kader was born in Cairo, Egypt, in 1941. He earned his BSc
in horticulture from the Faculty of Agriculture at Ain Shams University in
Cairo in 1959. He started as a medical student, but at 14 years of age he
found he couldn’t stomach human organs and blood; thus, he transferred
to the Faculty of Agriculture. After obtaining his BS from Ain Shams, he
moved to UC Davis, where he earned his MSc in vegetable crops in 1962
and his PhD in plant physiology in 1966 at the age of 25. After earning his
doctorate, Professor Kader returned to Egypt, where he held the position
of assistant professor in the Faculty of Agriculture at Ain Shams University
from 1966 to 1971. While there, he engaged in teaching and research on
postharvest horticulture and coauthored a classic postharvest textbook in
Arabic. He then became a lecturer and consultant in the Kuwait Institute
for Scientific Research from 1971 to 1972, before returning to UC Davis in
1972, first as an assistant researcher and later as an assistant, associate,
and full professor until his retirement in 2007. He held the title of emeritus
professor until his death. During his tenure at UC Davis, he served as chair-
man of the Department of Pomology from 1986 to 1991, a member of the

xxiii
D ed ic ati o n

campus academic planning council, and on numerous committees of the

UC system-wide Division of Agriculture and Natural Resources.
Dr. Kader received numerous awards. A listing of just a few of them
illustrates the breadth of his accomplishments. He was elected a fellow of
the American Society for Horticultural Sciences (ASHS) in 1986, later serv-
ing as president-elect in 1994–1995, president in 1995–1996, and chairman
of the board of directors in 1996–1997, as well as on the finance commit-
tee and as chair of the publications committee. Dr. Kader also received
awards for best research publications in 1978 and 1980 from the ASHS.
He was the chair of the program committee when UC Davis hosted the
22nd International Horticultural Congress in 1986. He attended every
controlled atmosphere conference, beginning with the second in 1977,
and convened the seventh edition of this conference in Davis in 1997. He
received from UC Davis the Outstanding Teaching Award in Extension
in 1989, the Award of Distinction from the College of Agricultural and
Environmental Sciences in 2000, the Alumni Citation of Excellence from
the Cal Aggie Alumni Association in 2000, and the Academic Senate’s
Distinguished Graduate Mentoring Award in 2003. He was also selected
as the Outstanding Horticulturist of 1997 by the Horticultural Research
Center at Laval University, Quebec, Canada. In April 2010, he received an
honorary doctorate degree from the University of Cartagena in Spain. In
2012, he was honored by the government of Malaysia for his outstanding
contributions to postharvest science, education, and extension.
As a member of several professional societies, Professor Kader served
on the editorial boards of Postharvest Biology and Technology, International
Journal of Postharvest Technology and Innovation, Postharvest News and
Information, and Tropical Science.
He cooperated with international organizations such as the Food and
Agriculture Organization (FAO) and United Nations Organization (UNO),
government programs such as the U.S. Agency for International Development
(USAID), and foundations such as the Gates Foundation. He worked with
many countries, including Saudi Arabia, Egypt, Syria, Iraq, India, Lebanon,
Mexico, Turkey, Morocco, Ghana, Sudan, Philippines, Thailand, Malaysia,
China, Chile, Jordan, and Kuwait. His laboratory and his home hosted a
continuing stream of visiting scientists from around the world. During his
university career, he trained 36 PhD students and more than 60 postdoc-
toral researchers who came from all over the world. Professor Kader served
for 18 years on the selected group of the Scientific Advisory Council of the
World Foods Logistics Organization, where he voluntarily supported the
industry all over the world.
Adel’s energy was also essential to the publication of the first and
subsequent editions of the companion text Postharvest Technology of
Horticultural Crops, third edition. More than 5700 copies in English have
been sold, and the text has also been translated into Spanish. When he

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 D ed ic ati o n

died, he was coordinating the writing of the fourth edition, his signature
book. He also served as author and editor of many publications, ­including
the popular Small-Scale Postharvest Handling Practices: A Manual for
Horticultural Crops, which he coauthored with Lisa Kitinoja and was trans-
lated into 11 additional languages. He was also associate editor of the book
Dates: Postharvest Science, Processing Technology and Health Benefits, which
was published after his death. Adel led the development of the Postharvest
Technology website (www.postharvest.ucdavis.edu), which has become
the premier place to find postharvest information and receives several mil-
lion page views annually.
In teaching and research, Adel was a wonderful colleague. He and
his students were particularly focused on understanding the physiology,
biochemistry, and technology of controlled and modified atmosphere stor-
age of fruits and vegetables. He held the highest ethical, professional, and
research standards for both himself and others, which challenged everyone
with whom he worked to perform to their highest possible level. In addition
to hundreds of peer-reviewed papers and popular articles describing his
research, Adel, with photographer Don Edwards, also produced hundreds
of high-quality slides demonstrating his research findings. These slides are
still an essential component of many of the presentations made by members
of the UC Davis postharvest team. We would like to remember his intellec-
tual integrity and his way of discussing and interacting with colleagues and
students. He was happy to share his knowledge, he had no secrets, and he
believed research and information should be shared by and with everyone.
He was happy to give a bibliographic reference, photos, PowerPoint presen-
tation, publications, and so on.
Adel was convinced that improved postharvest practices would not
only raise the quality, taste, and nutrition of fruits and vegetables in the
United States, but also improve food supply and farmers’ incomes in the
developing world. From the start of his career, he was constantly involved
in development activities. As a key player in the Agriculture Development
Strategy (ADS) project, which sought to bring the expertise of U.S. hor-
ticulturists to Egypt, he made postharvest handling a central theme. The
subsequent flourishing of horticulture and horticultural exports from his
home country can, at least in part, be attributed to his efforts.
After retirement, Professor Kader maintained a very active interest in
postharvest programs worldwide and frequently participated in seminars at
UC Davis and many international meetings, chaired the California Citrus
Quality Council and the research advisory board of the Produce for Better
Health Foundation, and was board director of the Postharvest Education
Foundation. He continued to do some consulting to raise funds for the UC
Davis Postharvest Endowment. For the past several years, Adel served as a
key player in the Global Horticulture Assessment, which laid the foundation
for the development of the Horticultural Collaborative Research Support

xxv
D ed ic ati o n

Program (Horticulture CRSP), which USAID awarded to UC Davis. As an

advisor during the writing of the proposal, and as a member of its inter-
national advisory board, he made significant contributions to the success
and direction of the program. In nearly every project in which Adel was
involved, he took care to empower others working alongside him, and it is
due to this foresight that many of these projects were, and will continue to
be, completed to fruition, to the benefit of numerous others.
With the sudden death of Adel Kader on December 10, 2012, the
postharvest and horticultural development community mourned the loss
of a leader, teacher, mentor, colleague, and friend. His big heart, which
he shared so willingly with everyone, finally failed him while traveling
home from a postharvest conference in South Africa. In August 2014, at
a general assembly during the International Horticultural Congress in
Brisbane, Australia, Professor Kader was awarded an International Society
for Horticultural Sciences (ISHS) fellow posthumously, for his outstand-
ing contribution to horticultural science in general and postharvest science
and technology in particular during his long and distinguished career.
Professor Kader was instrumental in providing anyone who had the
privilege of working with him the necessary tools to feel at home in the
scientific community and become a contributor to the body of knowledge of
plant science. He also provided everyone in horticulture, and postharvest
biology in particular, something more important—the reinforcement of a
personal code of ethics that is crucial for being a member of the human
race. His high level of professional conduct, the care he took in mentoring
his students, and his humble approach to life are what enabled him to have
such an impact, not only on the world of postharvest biology, but also on the
world in general. Last but not least, it can be said that if postharvest is the
religion, Dr. Kader is the bible.

Elhadi M. Yahia, Mikal E. Saltveit, and Sunil Pareek

xxvi
 Series Preface

The ‘Innovations in Postharvest Technology’ book series provides

updated and comprehensive information on the innovations and emerging
technologies in postharvest and processing of horticultural commodities,
as well as postharvest physiology, biochemistry, ripening, and engineer-
ing aspects. The series includes books on ripening physiology, biochemis-
try, treatments to enhance shelf life, chilling injury, fresh cut and minimal
processing, postharvest pathology, and physiological disorders. The series
also includes books on postharvest biology and technology of tropical, sub-
tropical, and temperate fruits of global importance, as well as vegetables and
spices. Books also cover several aspects of general interest, such as supply
chain management, postharvest technology status in various regions of the
world, analytical techniques, biotechnology, engineering, nondestructive
quality evaluation, and health effects. The books are aimed at food scientists,
postharvest researchers and industries, and graduate- and postgraduate-
level students.

xxvii
 Foreword

Horticultural crops have myriad uses in societies worldwide; not only are
fruits and vegetables critical components of the diet, but together with flow-
ers and ornamentals, they are pleasing aesthetic products that contribute
to human well-being. Edible products are a source of antioxidant vitamins
(A, C, and E), phenolics, carotenoids, phytonutrients, and dietary fiber that
are important health-promoting compounds in the human diet. The con-
tribution that these products make to human health has become increas-
ingly recognized and includes reduction of the incidences of degenerative
diseases as well as cardiovascular disease, hypertension, and cancers.
However, no amount of information about the “goodness” of any fruit or
vegetable is useful if it is not visually appealing or flavorful or does not meet
the quality expected by the consumer.
Access to horticultural products can vary greatly, with scarcity in
some areas and surplus in others. Estimates for losses of horticultural
crops vary widely, but it is interesting from survey research that losses of
fruits and vegetables are similar in the developing and developed worlds.
Yet, where those losses occur, between the “farm and fork,” can be mark-
edly different. In the developing world, losses are largely a result of fac-
tors such as lack of harvest, storage, and transport infrastructure and poor
marketing systems. In the developed world, losses are greater after harvest
because of stringent quality standards and factors such as excess produc-
tion, loss of product quality after harvest, and “plate waste” (unconsumed
food after purchase by consumers). Interestingly, the locavore movement,
or seeking locally grown food, often organic, in countries such as the United
States, can sometimes resemble the situation in developing countries, with

xxix
Fo re wo rd

small-scale and hobby farms lacking access to proper refrigeration and

appropriate marketing channels.
Selection for biological and physiological properties that ensure bet-
ter storage and transport capability of horticultural crops has often led
to products that are firm and slow to ripen, but with less sensory appeal,
such as relatively poor flavor and aroma. Also, the compromise between
quality and storage potential that exists for many fruits can result in their
harvest well before full-quality characteristics are attained in order to
maximize storage periods. Therefore, quality in the marketplace is often
at minimum acceptable levels. Together with selection of crops for yield
and uniformity of appearance, rather than consumer quality has resulted
in the paradox that consumers in developed countries have more choice
than any time in history, and yet dissatisfaction with food quality is high.
In part, this has led to the locavore movement because of the desire of con-
sumers to eat horticultural products that have not traveled long distances
and are perceived as fresher and healthier.
It is in this context that books such as Postharvest Ripening Physiology
of Crops provide an opportunity to summarize our understanding of the
complex interactions that result in ripening. Our ability to modify these
processes by breeding, whether traditionally or by manipulation of specific
genes by technology, requires ongoing research. In addition, understand-
ing of ripening processes has underpinned the development of existing
postharvest technologies and will be essential for development and imple-
mentation of newer technologies. Our understanding of ripening and
senescence processes in horticultural crops has progressed substantially
beyond the descriptive knowledge of the recent past. This progress has
been due to advances in omics technologies that are allowing identifica-
tion of genomic, proteomic, and metabolomic events that initiate and modu-
late these processes. The chapters in this book address these aspects, but
notably, morphological and physiological factors that affect ripening, and
thereby responses to postharvest environments, are also covered. Much
remains to be learned as the need to concurrently breed horticultural crops
with high-quality characteristics, while providing the ability to maintain
quality throughout storage, transport, and shelf life, and reduce waste, in
order to deliver high-quality crops with health benefits to the consumer,
will continue to increase.

Christopher B. Watkins

Cornell University, Ithaca, New York

xxx
 Preface

Ripening is an important aspect of the production and shelf life of fresh pro-
duce. Timing and stage of ripening affect the buying behavior of produce,
shelf life, quality, and nutraceuticals. Ripening physiology is complicated
and affected simultaneously by many factors. Looking at these facts, this
book provides information on the postharvest physiology, biochemistry, and
molecular biology of ripening. Quality, physiology, and molecular biology
of flower senescence are also discussed. Detailed information on advances
in respiration measurement, stomatal relations in postharvest, and factors
controlling postharvest water loss has been provided. Lysophospholipids
research is gaining popularity and provides information on ripening and
extending shelf life of horticultural produce. A detailed account on posthar-
vest quality in relation to lysophospholipids is also included.
This book is a comprehensive interdisciplinary reference source for
the various aspects of fruit ripening and postharvest behavior. It focuses on
the postharvest physiology and ripening overview of fruits and vegetables.
Flower senescence is equally important for the postharvest horticulture
industry, and chapters are included on the postharvest quality of ornamen-
tal plants and molecular biology of flower senescence. The share of colored
fruit in the total fruit production is quite significant, with the largest con-
tributing several billion dollars annually. This has encouraged scientists
to study the pigmentation mechanism in fruits. Various developments that
have taken place in the last decade with respect to identifying and altering
the function of ripening-related genes are described in this book. Taking
clues from studies in grape as a model fruit, we review a few case studies,

xxxi
Preface

and a detailed account of molecular regulation of fruit ripening, signal

transduction, and internal atmospheres in relation to fruit ripening is given.
Comparative proteomics is a useful tool to gain information on the
molecular events taking place during fruit maturation, in addition to find-
ing biotechnological strategies to improve horticultural traits, such as fruit
quality, shelf life, and yield. An overview of methods utilized in fruit pro-
teomics, as well as a global proteome and systems biology analysis of fruits
during ripening, is also presented. Potato is an important food crop of the
world; potato dormancy is associated with postharvest storage. The basics
of dormancy, its molecular and physiological basis, and methods to break
the dormancy are also discussed in detail in this book.
This book provides an overview of the most important metabolic path-
ways and genes that control volatile biosynthesis in model fruits, including
tropical, subtropical, and temperate fruits, with a special emphasis on fruit
ripening and the role of ethylene during this process. Also presented is
a brief description of the composition of volatiles in various fruit species
and a discussion of the influences of preharvest factors and postharvest
technologies on fruit aroma. A basis for product flavor, basic mechanisms
responsible for postharvest flavor change in fresh produce, and the poten-
tial impacts of various postharvest technologies on flavor are addressed.
I am sure that this book will serve as an important comprehensive
reference work for researchers, students, postharvest industries, and any-
one else who is involved in maintaining the postharvest quality of fresh
produce.

Sunil Pareek

Associate Professor (PHT)

Department of Agriculture & Environmental Sciences
National Institute of Food Technology Entrepreneurship and Management
(NIFTEM)
(Deemed University under Ministry of Food Processing Industries)
Haryana, India

xxxii
 Acknowledgments

I gratefully acknowledge the special collaborations and suggestions made

by the scientists from the Maharana Pratap University of Agriculture and
Technology (MPUAT), Udaipur, India; National Institute of Food Technology
Entrepreneurship and Management (NIFTEM), Kundli, Haryana, India;
and Indian Council of Agricultural Research (ICAR), New Delhi, India.
Much appreciation is expressed to Dr. Ajit Kumar, vice chancellor, NIFTEM;
Dr. K.L. Chadha, former deputy director general (Horticulture Sciences),
ICAR; Dr. S.K. Malhotra, horticulture commissioner, Government of India;
Dr. H.P. Singh, former deputy director general (Horticulture Sciences),
ICAR; and Dr. S.L. Mehta, former deputy director general (Education),
ICAR, and vice chancellor, MPUAT, Dr. R. Paliwal, professor at Rajasthan
Agricultural Research Institute, Jaipur, India, and Dr. J.G. Varshney, head,
Department of Agriculture and Environmental Sciences, NIFTEM, Kundli,
India. The financial support by the ICAR under the National Agricultural
Research Project is deeply appreciated.
I am grateful to each of the authors for their participation, prompt-
ness, patience, and cooperation, as well as many others for their contribu-
tions, advice, and encouragement in the development of this book. I would
like to thank Ashley Weinstein and Stephen Zollo (CRC/Taylor & Francis)
for their support and encouragement in the preparation of the book ­proposal
and manuscript.
With my head stopped, I feel a paucity of words to express my
humble sense of regard to my parents, Mrs. Sushila and Dr. R.G. Pareek.
Finally, I think words are insufficient to express the feelings of my heart to

xxxiii
Ac k n o w l e d g m e n t s

acknowledge my better half, Dr. Shilpi, and son, Mr. Sabhya, who under-
went all sorts of hardships and sufferings to support my spirit and endeavor
at every step.

xxxiv
 Editor

Dr. Sunil Pareek earned his PhD in horticulture

(PHT) from Rajasthan Agricultural University,
Bikaner, India. He joined Maharana Pratap University
of Agriculture and Technology (MPUAT), Udaipur,
India, in 2005 as an Assistant Professor (PHT) in the
Department of Horticulture, Rajasthan College of
Agriculture, MPUAT, Udaipur, India and worked there
up to August, 2015. Presently he is working as associ-
ate professor (PHT) in the Department of Agriculture
and Environmental Sciences, National Institute of
Food Technology Entrepreneurship and Management (NIFTEM), Kundli,
India.
He is involved in teaching UG, PG, and PhD students, with a spe-
cial focus on postharvest physiology, technology, and processing of fruits.
He has guided several MSc and PhD students in PHT. He has executed
research projects supported by the Indian Council of Agriculture Research,
World Bank, Ministry of Tribal Development, and Rajasthan Mission on
Livelihood. Currently he is a co-consortium principal investigator (co-PI)
of the National Agricultural Innovation Project (NAIP) on underutilized
fruits, PI of the All India Coordinated Research Project on Tuber Crops, PI
of the Processed Products Scheme, and co-PI of the Integrated Farming
Systems Project.
Dr. Pareek has generated many technologies for the extension of the
shelf life of indigenous fruits and their processed products. Presently, he is
involved in a research program on applications of postharvest physiology of

xxxv
Ed i t o r

fruits to maintain their quality during storage. He standardized and com-

mercialized the browning free pulp extraction and preservation technol-
ogy for sugar apple fruit. Dr. Pareek has published more than 40 papers,
40 ­presentations in national and international seminars and conferences,
6 books, 3 manuals, 6 technical bulletins, and 40 popular articles, and he
has several book chapters to his credit. He is the founder life member of
the Indian Society of Arid Horticulture and a member of several scientific
societies. He is on the reviewers’ panel of 12 international journals of repute
and also the editor, associate editor, or editorial board member of 11 inter-
national journals. He is the recipient of the University Outstanding Services
Award 2013, Young Scientist Award 2012, HS Mehta Young Scientist Award
2012, Fellow Award of the Confederation of Horticultural Associations of
India, and a few best poster awards.

xxxvi
 Contributors
Sasan Aliniaeifard
Department of Horticulture
College of Abureyhan
University of Tehran
Tehran, Iran
Domingos P.F. Almeida
Instituto Superior de Agronomia
Universidade de Lisboa
Lisbon, Portugal
Jane Ambuko
Department of Plant Science and
Crop Protection
College of Agriculture and
Veterinary Services
University of Nairobi
Nairobi, Kenya
Fernanda F. Araujo
Department of Plant Physiology
Federal University of Viçosa
Viçosa, Minas Gerais, Brazil
Ajay Arora
Division of Plant Physiology
ICAR-Indian Agricultural Research
Institute
New Delhi, India
Jose G. Barbosa
Department of Phytotechnology
Federal University of Viçosa
Viçosa, Minas Gerais, Brazil
Michael A. Campbell
School of Science
Penn State Erie, The Behrend
College
Erie, Pennsylvania
Bruno G. Defilippi
Instituto de Investigaciones
Agropecuarias (CRI La Platina)
La Pintana, Santiago, Chile
xxxvii
Co ntributo rs
Sergio Tonetto de Freitas
Brazilian Agricultural Research
Corporation–Embrapa
Embrapa Tropical Semi-Arid
Petrolina, Pernambuco, Brazil
Cassandro Vidal Talamini do
Amarante
Department of Agronomy
Santa Catarina State University
Lages, Santa Catarina, Brazil
Fernando L. Finger
Department of Phytotechnology
Federal University of Viçosa
Viçosa, Minas Gerais, Brazil
Charles F. Forney
Atlantic Food and Horticulture
Research Centre
Agriculture and Agri-Food Canada
Kentville, Nova Scotia, Canada
Christoph Gehring
Division of Biological and
Environmental Sciences and
Engineering
King Abdullah University of
Science and Technology
Thuwal, Kingdom of Saudi Arabia
Orianne Gudenschwager
Instituto de Investigaciones
Agropecuarias (CRI La Platina)
La Pintana, Santiago, Chile
M. Mofazal Hossain
Department of Horticulture
Bangabandhu Sheikh Mujibur
Rahman Agricultural University
Gazipur, Bangladesh
xxxviii
Kazuo Ichimura
NARO Institute of Floricultural
Science
National Agriculture and Food
Research Organization
Fujimoto, Tsukuba, Japan
Emrul Kayesh
Department of Horticulture
Bangabandhu Sheikh Mujibur
Rahman Agricultural University
Gazipur, Bangladesh
Ola Lasekan
Faculty of Food Science and
Technology
University Putra Malaysia
Serdang, Malaysia
Claudius Marondedze
Department of Biochemistry
University of Cambridge
Cambridge, UK
Uulke van Meeteren
Department of Plant Sciences
Wageningen University
Wageningen, the Netherlands
Elizabeth J. Mitcham
Department of Plant Sciences
University of California
Davis, California
Nora L. Olsen
Twin Falls Research and Extension
Center
University of Idaho
Twin Falls, Idaho

Willis O. Owino
Department of Food Science and
Technology
Faculty of Agriculture
Jomo Kenyatta University of
Agriculture and Technology
Nairobi, Kenya
Rakesh Pandey
Division of Plant Physiology
ICAR-Indian Agricultural Research
Institute
New Delhi, India
Sunil Pareek
Department of Horticulture
Rajasthan College of Agriculture
Maharana Pratap University of
Agriculture and Technology
Udaipur, Rajastha, India
and
Department of Agriculture &
Environmental Sciences
National Institute of Food
Technology Entrepreurship and
Management (NIFTEM)
Kundli, Sonepat, Haryana, India
Vijay Paul
Division of Plant Physiology
ICAR-Indian Agricultural Research
Institute
New Delhi, India
Mikal E. Saltveit
Mann Laboratory
Department of Plant Sciences
University of California
Davis, California
Co ntributo rs
Lingfei Shangguan
College of Horticulture
Nanjing Agricultural University
Nanjing, China
Kenichi Shibuya
NARO Institute of Floricultural
Science
National Agriculture and Food
Research Organization
Fujimoto, Tsukuba, Japan
Tania P. Silva
Department of Phytotechnology
Federal University of Viçosa
Viçosa, Minas Gerais, Brazil
Jeffrey C. Suttle
USDA-ARS Northern Crop Science
Laboratory
Fargo, North Dakota
Ludivine Thomas
Bioscience and Bioengineering
Core Facility
King Abdullah University of
Science and Technology
Thuwal, Kingdom of Saudi Arabia
Peter M.A. Toivonen
Pacific Agri-Food Research Center
Agriculture and Agri-Food Canada
Summerland, British Columbia,
Canada
Christopher B. Watkins
School of Integrative Plant
Science—Horticulture Section
Plant Science Building
Cornell University
Ithaca, New York
xxxix
Co ntributo rs
Aloysius Wong
Division of Biological and
Environmental Sciences and
Engineering
King Abdullah University of
Science and Technology
Thuwal, Kingdom of Saudi Arabia
xl
Elhadi M. Yahia
Human Nutrition Program
Faculty of Natural Sciences
Autonomous University of
Queretaro
Juriquilla, Queretaro, Mexico
Chapter 1

Ripening Physiology:
An Overview
Sunil Pareek 1,2
1Maharana Pratap University of Agriculture and
Technology, Udaipur, Rajasthan, India
2National Institute of Food Technology Entrepreneurship
and Management, Ministry of Food Processing
Industries, Kundli, Sonepat, Haryana

Abstract 2
1.1 Introduction 2
1.2  Climacteric Phenomenon 4
1.3 Physicochemical and Metabolic Changes 12
1.3.1  Color Changes 13
1.3.2  Sugar Changes 14
1.3.3  Organic Acid Changes 16
1.3.4  Flavor and Aroma Changes 18
1.3.5  Cell Wall and Textural Changes 22
1.3.6  Physiological Changes 28
1.4  Conclusions and Future Perspectives 33
References 33

1
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Abstract
Ripening is an important event in the fruit life cycle and from the consum-
ers’ point of view. Mature fruit undergo ripening, which is a coordinated
process typically involving many changes, such as pigmentation, flavor,
aroma, respiration, ethylene, texture, softening, sugar, and organic acid.
Physiologists typically classified fruits into climacteric and nonclimacteric
categories on the basis of their respiration peaks and internal ethylene
concentrations. This chapter reviews the climacteric and nonclimacteric
ripening of fruits and physiological and metabolic changes during ripen-
ing. In climacteric fruit, components responsible for the production of cli-
macteric ethylene have been identified. Less progress has been made on
nonclimacteric fruit; still, knowledge is poor to classify many fruits into
climacteric or nonclimacteric. A comprehensive climacteric classification
of fruits is given in this chapter. Textural changes and the role of enzymes
are also reviewed.

1.1 Introduction
Ripening is the composite of the processes that occur from the latter stages
of growth and development through the early stages of senescence and that
result in characteristic aesthetic and food quality, as evidenced by changes
in composition, color, texture, or other sensory attributes. Fruit ripening
is a complex process that undergoes dramatic changes mainly involving
flavor, color, and texture. It is a controlled process where the cellular com-
munication is important, as well as the influence of plant growth regulators
and environmental signals (Cruz-Hernandez and Paredes-Lopez, 2012).
Ripening could also be defined as a physiological process that is geneti-
cally programmed and comprises several physical, chemical, and biochemi-
cal changes that render fruit attractive and palatable (Lelievre et al., 1997;
Giovannoni, 2001).
One early view of ripening was that it is due to entirely to a series
of catabolic reactions associated with changes in membrane permeabil-
ity and a decrease in the structural integrity of the cell, resulting in the
release or activation of hydrolytic enzymes. This view is now clearly
untenable since genetic studies suggest that the expression of specific
genes is also required for ripening. It is supported by biochemical evi-
dence that shows there are changes in specific mRNAs and the de novo
synthesis of protein during ripening. Changes in gene expression are
both positive and negative. They involve genes in the nucleus and plas-
tids, and expression of at least some of them is confined to ripening fruit
tissues. Thus, fruit ripening is a highly controlled and programmed

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

developmental event, involving the coordination of a multitude of meta-

bolic changes (Seymour et al., 2002) and the activation and inactivation
of various genes (Clendennen and May, 1997; Medina-Suarez et al., 1997;
Drury et al., 1999; Itai et al., 2000), leading to various biochemical and
physiological changes within the tissue.
At ripening, fruits undergo many changes (Table 1.1), which, although
variable among species, generally include modification of cell wall ultra-
structure and texture, conversion of starch to sugars, alterations in pig-
ment biosynthesis and accumulation, and heightened levels of flavor and
aromatic volatiles (Brady, 1987).

Table 1.1  Changes Occurring in Fruit Ripening

Changes Events
Biochemical
Color Loss of chlorophyll
Dismantling of photosynthetic apparatus
Synthesis and accumulation of pigments
Texture Solubilization of pectin and cellulose
Starch degradation
Changes in protein content
Hydration of cell walls
Cell wall enzyme activity
Flavor and aroma Accumulation of sugars and organic acids
Production of volatiles
Alcohol ester synthesis
Metabolic
Control of pathways Increase in respiration
Ethylene synthesis
Changes in metabolism of starch and organic
acids
Altered regulation of existing metabolic pathways
Molecular
Gene expression Ripening-specific mRNA synthesis
Small and interference RNA appearance
Disappearance of mRNAs
Protein expression Synthesis of de novo ripening-specific proteins
Disappearance of proteins

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

1.2  Climacteric Phenomenon

Fruits can be broadly classified as climacteric or nonclimacteric, depend-
ing on whether or not a fruit exhibits a peak in respiration and ethylene
production during ripening. Climacteric fruits are characterized by a tran-
sient increase in ethylene synthesis and respiration at an early stage of
ripening. The peak of the ethylene production rate is proportional to the
peak respiration rate. An increased rate of respiration that occurs at an
early stage in ripening is termed climacteric, the term currently used to
describe the totality of events occurring during the ripening of climacteric
fruit. The study of the respiratory curve of climacteric fruits at a suitable
temperature shows a decreasing trend to the lowest value, termed the pre-
climacteric minimum, followed by a rise in respiration to the climacteric
peak and the subsequent postclimacteric decline in the rate of respiration
(Figure 1.1). During the ripening of climacteric fruits, the increase in ethyl-
ene and respiration is an early event (Figure 1.2). In many cases, occurring
prior to any changes in color or texture, this peak respiration corresponds
with eating ripeness, as has been observed in avocado, banana, cherimoya,
and mango. Fruit softening, color changes, development of taste and flavor,

Climacteric peak
Carbon dioxide production

Post-climacteric
decline
Climacteric
rise

Pre-climacteric
minimum

Non-climacteric pattern

Time

Figure 1.1  Climacteric pattern of respiration.

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Climacteric fruit

180 Breadfruit

Cherimoya
160

140
ml O2 or CO2/Kg-Hr

120

100

Mango
80

60

Fig
40

20 Tomato
Apple

0 2 4 6 8 10 12 14 16 18
Time units

Figure 1.2  Respiration rate of some climacteric fruits.

and a number of other parameters of the ripening process are associated

with the climacteric cycle. Climacteric is therefore defined as a period in
the ontogeny of fruit during which a series of biochemical changes are
initiated by autocatalytic production of ethylene, making the change from
growth to senescence and involving an increase in respiration, leading to
ripening of the fruit (Payasi and Sanwal, 2005). Nonclimacteric fruit does

5
P OST H AR V EST RI P ENING P H Y SIOLOG Y

not show any increase in respiration and ethylene synthesis during ripen-
ing (Figure 1.3). In fact, nonclimacteric fruits show decline in their respira-
tion rate and ethylene production throughout the ripening process.
Climacteric events are mainly regulated by the gaseous phytohor-
mone ethylene, which is also involved in the decrease in flesh firmness
typical of many economically relevant crops, such as tomato and peach
(Prinsi et al., 2011). On the other hand, ripening of nonclimacteric fruits
such as pepper, citrus, and strawberry is ethylene independent, although
similar major visual, texture, flavor, and metabolic changes occur, as in cli-
macteric fruits. Many of the changes have been mainly characterized in
climacteric ripening fruits, whereas nonclimacteric fruit ripening is still
poorly understood. A list of climacteric and nonclimacteric fruit updated
from Biale and Young (1981), Kays (1991), and Watkins (2002) is shown
in Table  1.2. Interestingly, this physiological behavior is not linked to

Nonclimacteric fruit

30

Strawberry
ml O2 or CO2/Kg-Hr

Grape
20
Pineapple

Cherry

10

Lemon

0
0 1 2 3 4 5 6 7 8 9 10
Time units

Figure 1.3  Respiration rate of some nonclimacteric fruits.

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.2  Climacteric and Nonclimacteric Classification of Fruit

Common Name Scientific Name Reference
Climacteric
Acerola Malpighia emarginata DC. Carrington and King
(2002)
Apple Malus pumila Mill. Biale (1960)
Apricot Prunus armeniaca L. Biale (1960)
Asian pear Pyrus serotina Rehder Downs et al. (1991)
Araza Eugenia stapitata Hernandez et al. (2007)
McVaugh
Atemoya Annona squamos x Brown et al. (1988)
Annona cherimola
Avocado Persia americana Mill. Biale (1960)
Bael Aegle marmelos (L.) Corr. Roy (1975)
Serr.
Banana Musa spp. L. Biale (1960)
Biriba Rollinia deliciosa Safford Biale and Barcus (1970)
Bitter melon Momordica charantia L. Kays and Hayes (1978)
Blackberry Rubus spp. L. Walsh et al. (1983)
Blueberry, Vaccinium angustifolium Ismail and Kender (1969)
lowbush Ait.
Blueberry, Vaccinium corymbosum L. Ismail and Kender (1969)
highbush
Blueberry, Vaccinium ashei Reade Lipe (1978)
rabbiteye
Breadfruit Artocarpus altilis Biale and Barcus (1970)
Caimito Pouteria caimito Malik et al. (2014)
Caja Spondias mombin L. Sampaio et al. (2007)
Camu-camu Myrciaria dubia Kunth Hernandez et al. (2009)
McVaugh
Canistel Pouteria campechiana Malik et al. (2014)
Cantaloupe Cucumis melo L. Lyons et al. (1962)
Cape gooseberry Physalis peruviana L. Trinchero et al. (1999)
Cherimoya Annona cherimola Mill. Biale (1960)
Chili plum Spondias purpurea var. Sampaio et al. (2007)
Lutea
Corossol sauvage Rollinia orthopetala A. DC. Biale (1976)

(Continued )

7
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 1.2  Climacteric and Nonclimacteric Classification of Fruit

Common Name Scientific Name
Dabai Camarium odontophyllum
Miq.
Date palm Phoenix dactylifera L.
Durian Durio zybethinus Murray
Feijoa Feijoa sellowiana O. Berg.
Fig Ficus carica L.
Giant granadilla Passiflora quadrangularis
Golden apple Spondias dulcis Forst. syn.
Spondias cytherea Sonn.
Golden berry Physalis peruviana L.
Guava Psidium guajava L.
Guava, ‘purple Psidium littorale var.
strawberry’ longipes (O. Berg.) Fosb.
Guava, Psidium rittorale Raddi
‘strawberry’
Guava, ‘yellow Psidium rittorale var.
strawberry’ littorale Fosb.
Honeydew melon Cucumis melo L. Inodorus
group
Jackfruit Artocarpus heterophyllus
Lam.
Jujube, Chinese Ziziphus jujuba Mill.
Jujube, Indian Ziziphus mauritiana Lamk.
Kiwifruit, Chinese Actinidia deliciosa (A.
gooseberry Chev) C.F. Liang et A.R.
Ferguson var. deliciosa
Lucuma Ponteria lucuma (Ruiz and
Pav.) Kuntze
Mammey apple Mammea americana L.
Mango, African Irvingia gabonensis Bailon
& Irvingiaceae
Mango, common Mangifera indica L.
Mangosteen Garcinia mangostana L.
Monstera Monstera deliciosa
Nance Byrsonima crassifolia (L.)
Kunth
Reference
Ding and Tee (2011)
Abbas and Ibrahim (1996)
Ketsa and Daengkanit
(1998)
Biale (1960)
Marei and Crane (1971)
Malik et al. (2014)
Graham et al. (2004)
Trinchero et al. (1999)
Akamine and Goo (1979)
Akamine and Goo (1979)
Akamine and Goo (1979)
Akamine and Goo (1979)
Pratt and Goeschl (1968)
Selvaraj and Pal (1989)
Kader et al. (1982)
Pareek and Yahia (2013)
Pratt and Reid (1974)
Yahia (2004)
Akamine and Goo (1978)
Aina and Oladunjoye
(1993)
Biale (1960)
Paull and Ketsa (2004)
Malik et al. (2014)
Velasquez de Klimo
(2001)
(Continued )

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.2  Climacteric and Nonclimacteric Classification of Fruit

Common Name Scientific Name Reference
Papaw Asimina triloba (L.) Dunal. Biale (1960)
Papaya Carica papaya L. Biale (1960)
Passion fruit, purple Passiflora edulis Sims Biale (1960)
Passion fruit, Passiflora edulis f. Malik et al. (2014)
yellow flavicarpa
Peach Prunus persica (L.) Batsch Biale (1960)
Pear, Chinese Pyrus breschneideri R. Tian et al. (1987)
Pear, European Pyrus communis L. Biale (1960)
Persimmon Diospyros kaki L. Reid (1975)
Plum Prunus americana Marsh. Biale (1960)
Raspberry Rubus idaeus L. Burdon and Sexton (1990)
Sapodilla Manilkara achras (Mill.) Malik et al. (2014)
Sapote Casimiroa edulis Llave Biale (1960)
Sapote, black Diospyros digyna Jacq. Yahia (2004)
Sapote, mamey Pouteria sapota Jacq. H.E. Yahia (2004)
Moore & Stearn
Sapote, white Casimiroa edulis Llave & Yahia (2004)
Lex
Saskatoon Amelanchier alnifolia Nutt. Rogiers et al. (1998)
Soursop Annona muricata L. Biale and Barcus (1970)
Sweetsop, sugar Annona squamosa L. Brown et al. (1988)
apple
Tomato Lycopersicon esculentum Biale (1960)
Mill.
Nonclimacteric
Achachairu Garcinia humilis (Vahl) Durate (2011)
C.D. Adam
Asian pear Pyrus serotina Rehder Downs et al. (1991)
Blackberry Rubus spp. L. Perkins-Veazie et al.
(2001)
Cacao Theobrama cacao L. Biale and Barcus (1970)
Cactus pear Opuntia amyclaea Tenore Lakshminarayana and
Estrella (1978)
Carambola Averrhoa carambola L. Lam and Wan (1983)
Cashew Annacardium occidentale L. Biale and Barcus (1970)
(Continued )

9
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 1.2  Climacteric and Nonclimacteric Classification of Fruit

Common Name Scientific Name Reference
Cherry, sour Prunus cerasus L. Blanpied (1972)
Cherry, Surinam Eugenia uniflora Malik et al. (2014)
Cherry, sweet Prunus avium L. Biale (1960)
Cranberry Vaccinium macrocarpon Ait. Kader (2002)
Cucumber Cucumis sativus L. Biale (1960)
Grape Vitis vinifera L. Biale (1960)
Grapefruit Citrus paradise Macf. Biale (1960)
Gumichama Eugenia brasiliensis Malik et al. (2014)
Indian gooseberry Emblica officinalis Gaertn. Pareek and Kitinoja
(2011)
Jaboticaba Myrciaria cauliflora (Mart.) Mota et al. (2002)
O. Berg
Java plum Syzygium cuminii (L.) Skills Akamine and Goo (1979)
Lemon Citrus jambhiri Lush. Biale (1960)
Litchi (Lychee) Litchi chinensis Sonn. Akamine and Goo (1979)
Longan Dimocarpus longan Lour. Zhao et al. (2005)
Loquat Eriobotrya japonica Lindl. Pareek et al. (2014)
Malay apple Syzygium malaccense Malik et al. (2014)
Mandarin Citrus reticulata Blanco Reid (1975)
Mountain apple Syzygium malaccense (L.) Akamine and Goo (1979)
Merrill & Perry
Olive Olea europeae L. Maxie et al. (1960)
Orange Citrus sinensis (L.) Osb. Biale (1960)
Pepper Capsicum annuum L. Saltveit (1977)
Pineapple Ananas comosus (L.) Merr. Biale (1960)
Pitanga Eugenia uniflora L. Santos et al. (2006)
Pitaya Hylocereus spp. Nerd et al. (1999)
Pitaya, yellow Selenicereus megalanthus Nerd and Mizrahi (1999)
Scum. Ex Vaupel
Pomegranate Punica granatum L. Ben-Arie et al. (1984)
Pummelo Citrus grandis Malik et al. (2014)
Rambutan Nephelium lappaceum L. Mendoza et al. (1972)
Raspberry Rubus idaeus L. Perkins-Veazie and
Nonneeke (1992)
Red bayberry Myrica rubra Sieb. Zucc. Joyce and Li (2003)
(Yang mei)
(Continued )

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.2  Climacteric and Nonclimacteric Classification of Fruit

Common Name Scientific Name Reference
Rose apple Syzygium jambos (L.) Alston Akamine and Goo (1979)
Star apple Chysophyllum cainito L. Pratt and Mendoza (1980)
Strawberry Fragaria x ananassa Duch. Biale (1960)
Strawberry, wild Fragaria vesca L. Nam et al. (1999)
Surinam cherry Eugenia euniflora L. Akamine and Goo (1979)
Tamarind Tamarindus indica Yahia (2004)
Tomato (nor-, rin-, Lycopersicon esculentum Thompson et al. (1999)
cnr-) Mill.
Tree tomato, Cyphomandra betacea Pratt and Reid (1976)
Tamarillo (Cav.) Sendtu
Watermelon Citrullus lanatus (Thumb.) Elkashif et al. (1989)
Mansf.
Wax apple Syzygium samarangense Akamine and Gao (1979)
(Blume) Merr. and L.M.
Perry)

taxonomic groups. Species belonging to the same family, such as tomato

and pepper (Solanaceae), display a distinct response to ethylene. Thus,
tomato is a climacteric fruit while pepper is not (Palma et al., 2011). With
other fruit, such as kiwifruit, a hybrid ripening pattern is seen, with most
of the ripening changes occurring in the absence of any detectable rise in
ethylene and CO2 production; a climacteric response occurs only toward
the end of ripening. Exposure to exogenous ethylene promotes ripening of
kiwifruit, but if exposure to ethylene is insufficient or fruit are too imma-
ture, then removal of ethylene results in nonclimacteric behavior.
White (2002), while reviewing fruit development and ripening, pro-
vides evidence from biochemical and genetic studies that both ethylene-
dependent and ethylene-independent regulatory cascades control the
development of tomato fruit. Hence, although the nonripening tomato
mutants ripening-inhibitor (RIN ) and non-ripening (NOR) do not produce
autocatalytic ethylene or ripen in the presence of exogenous ethylene, they
do display signs of ethylene sensitivity and ethylene-inducible expression
of several genes. Thus, it is likely that RIN and NOR participate in ethylene-
independent regulatory cascades during the early stages of fruit ripening
(White, 2002). Jim Giovannoni and colleagues have isolated the RIN and
NOR genes by positional cloning strategies (Moore et al., 2002; Vrebalov
et al., 2002). LeMADS-RIN encodes a member of the MADS-box family of
transcription factors. Homologues of LeMADS-RIN are expressed during
the ripening of other fruit, including strawberry, which might indicate a

11
P OST H AR V EST RI P ENING P H Y SIOLOG Y

common (ethylene-independent) function in the ripening of both climac-

teric and nonclimacteric fruit.
Several papers describe the role of ethylene in the ripening of climac-
teric fruit (Alexander and Grierson, 2002; Klee, 2002; Moore et al., 2002;
Payasi and Sanwal, 2009; Bapat et al., 2010; Bouzayen et al., 2010; Pech
et al., 2012, 2013). These concentrate on the elucidation of biochemical and
genetic signaling cascades that impact the development and ripening of
tomato fruit. The isolation of transcription factors NOR and LeMADS-RIN,
which participate in ethylene-independent signaling in tomato, and the dis-
covery that a homologue of the RIN gene is expressed in nonclimacteric
fruit have suggested that the common regulatory cascades may operate
in all fruits. This knowledge could enable generic strategies to manipulate
the ripening of any fruit (White, 2002). Such work has been complemented
by transcriptional profiling during the development and ripening of both
climacteric and nonclimacteric fruit (Aharoni and O’Connell, 2002; Moore
et al., 2002; Seymour et al., 2002), which may disclose more common regu-
latory elements.
Nevertheless, the timing of the climacteric syndrome in nonclimac-
teric fruit is not related to the ripening period per se. In grape, it occurs
at veraison (Chervin et al., 2004), and in citrus, in young immature fruit
(Katz et al., 2004). In strawberries, ethylene production starts to increase
once the fruit reaches the red ripe stage, but not before (Lannetta et al.,
2006). It is now considered that some aspects of the ripening of noncli-
macteric fruit are regulated by ethylene, and in climacteric fruit, some
ripening pathways are independent of ethylene action (Pech et al., 2013).
Differential cross talk between ethylene and other phytohormones prob-
ably operates in each type of fruit (refer to Chapter 12 for details) (Pech
et al., 2008; Paul et al., 2012).

1.3 Physicochemical and Metabolic Changes

Ripening changes involve a multiplicity of biochemical, metabolic, and
molecular changes that affect the cell compartments (Table 1.1): they have
been shown to be related to alterations in specific enzymes or complete
pathways. Color changes, for example, are due to alterations in the chloro-
phyll and pigment content of the plastids (Carrilo-Lopez et al., 2003; Zhang
et al., 2006). Softening is brought about by alteration in cell wall metabolism
and is due to a partial solubilization of pectin and cellulose; starch degra-
dation may contribute to a change in texture (Carrilo-Lopez et al., 2002).
Alterations in the metabolism of organic acids and the generation of volatile
compounds that produce aroma are common. It is clear that the various
changes associated with ripening take place in different parts of the cell
and are highly coordinated.

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

1.3.1  Color Changes

Changes in fruit color typically involve the destruction of chlorophyll to
reveal other pigments already present and may also involve the synthesis
of additional pigments (Table 1.3). More than 25 years ago, Von Loesecke
revealed that the yellowing of banana peels during ripening is essentially
an unmasking of preexisting carotenoids as chlorophyll is broken down,
although more recent work has shown that subtle changes do occur in the
various carotenoids present. In tomato, carotenoid synthesis accompa-
nies the loss of chlorophyll, while in fruits that turn purple, blue, or black,
anthocyanins are synthesized as chlorophyll is depleted. In tomato, where
this process has been well studied and represents the transformation of
the chloroplast into a chromoplast, carotenoid synthesis has been shown
to be light dependent (Giovannoni, 2001). In some fruits, ripening may also
involve a change in the fruit surface waxes, altering the bloom or shininess
of the fruit (Carrington, 2011). Table 1.4 provides the carotenoid concentra-
tions in various fruits.
Conversion of chloroplast-rich photosynthetic fruit to a chromoplast
and nutrient-rich, nonphotosynthetic fruit is essential for the development

Table 1.3  Carotenoid Derivatives Synthesized during Ripening in Different

Fruits
Carotenoid Derivative Fruits
Lycopene Tomato, watermelon, guava, papaya, grapefruit
β-Carotene Cantaloupe melon, mango, pumpkin, apricot
Zeaxanthin Orange pepper, citrus fruit, persimmon
Capsanthin Red pepper
Capsorubin Red pepper

Table 1.4  Range of Carotenoid Concentration in Some Fruits

Carotenoid Concentration
Fruit (mg 100 g−1) Reference
Apricot 0.1–4 Kurz et al. (2008)
Tomato 10.5–27.8 Ilahy et al. (2011)
Citrus species 0.02–5 Fanciullino et al. (2008)
Papaya 1.5–3 De Souza et al. (2008)
Loquat 0.1–2 Zhou et al. (2007)
Pepper 0.2–20 Topuz and Ozdemir (2007)
Watermelon 3–7 Perkins-Veazie et al. (2001)

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

of storage compartments (i.e., chromoplasts) with increased capacity to

accumulate large quantities of the lipophilic carotenoids (for more details,
refer to Egea et al., 2010). During the chloroplast-to-chromoplast transition,
specific carotenoid biosynthesis genes are expressed. The first step in carot-
enoid biosynthesis corresponds to the condensation of two geranylgera-
nyl diphosphate molecules into phytoene, which is catalyzed by phytoene
synthase (PHY ). In tomato fruit, two PHY genes are expressed. Phytoene
synthase 1 (PHY1) is highly expressed in ripening fruit and is responsible
for the formation of chromoplastic carotenoids, while phytoene synthase 2
(PHY2), which is responsible for the formation of chloroplastic carotenoids,
is expressed exclusively in green tissues, and therefore makes no contri-
bution to carotenoid biosynthesis in ripening fruit (Fraser et al., 1999). In
tomato, the isoprenoid biosynthetic pathway gives rise to carotenoids, such
as β-carotene and lycopene, gibberellins, quinines, and sterols. This path-
way has been well researched since its manipulation impacts not only the
organoleptic qualities of fruit, but also their contribution to human health.
Other red and purple pigments of the type seen in grapes and boy-
senberries are anthocyanins, which are products of the phenylpropanoid
pathway. Anthocyanin pigments are water soluble, synthesized in the cyto-
sol, and localized in the vacuole. Their basic ring structure can be modified
by hydroxylation, methylation, or glycosylation, and their specific color is
modified by pH, metal ions, and co-pigments to produce the subtlety of col-
ors seen in nature. Anthocyanin content varies significantly in a range of
fruits (Table 1.5).
The accumulation of anthocyanins is regulated by transcription fac-
tors of two classes (R2R3 MYB and basic helix loop helix), regulatory pro-
teins that coordinate gene expression of the whole phenylpropanoid pathway.
In fruit, this regulation system has been well characterized in grape and
apple. In white berry grapes, VvMYBA2 is inactivated by mutations in the
coding region and VvMYBA1 has a retrotransposon in the promoter and is
not transcribed (Kobayashi et al., 2004; Walker et al., 2007). In apple fruit,
a mini-­satellite repeat structure in the promoter region of the MYB10 gene
upregulated the expression of this regulatory gene, which increased the level
of anthocyanin throughout the plant, producing a fruit with striking red color
throughout the flesh (Espley et al., 2009) (for details, refer to Chapter 9).

1.3.2  Sugar Changes

During fruit ripening, sugar levels within fruit tend to increase (Whiting,
1970), due to either increased sugar importation from the plant or the mobi-
lization of starch reserves within the fruit, depending on the fruit type and
whether it is ripened on or off the plant. The sugar or sugar alcohol, deliv-
ered to the fruit, is converted to starch (e.g., mango, banana, kiwifruit),

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.5  Anthocyanin Content in Common Fruits

Fruit Anthocyanin (mg 100 g−1 FW)
Acai 53.6
Acerola 22.6
Apple, Fuji 0.7
Apple, Gala 1.1
Apple, Golden 0.0
Apple, Granny Smith 0.0
Apple, Red Delicious 3.8
Avocado 0.3
Blackberry 90.6
Blueberry 141.0
Cherry 27.7
Cranberry 85.5
Currant, black 154.8
Currant, red 75.0
Currant, white 0.0
Elderberry 485.3
Grape, Concord 65.6
Grape, red 44.0
Grape, green 0.0
Kiwifruit 0.0
Melon 0.0
Nectarine and peach 1.8
Pear 12.2
Pineapple 0.0
Plum, red 6.98
Plum, black 39.7
Plum, yellow 0.3
Raspberry 40.9
Strawberry 23.8
Watermelon 0.0
Note: FW, Fresh Weight.

stored as reducing sugar (e.g., tomato, strawberry), or stored as sucrose

(e.g., wild tomato, watermelon, grape), or may even be converted to ­lipids
(e.g., olive). Accumulation of sucrose, glucose, and fructose in fruits
such as melon, watermelon (Brown and Summers, 1985), strawberry

15
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 1.6  Starch Mobilizing Enzymes and Their Response in Fruits

Enzymes Mode of Action
α-Amylase Hydrolyzes the α (1–4) linkages of amylase at random to
produce a mixture of glucose and maltose
β-Amylase Attacks the penultimate linkage and releases only maltose
Starch Hydrolyzes the terminal α (1–4) linkage to give glucose-1-
phosphorylase phosphate, which can be converted to glucose-6-
phosphate by the action of glucose phosphate mutase
α-1,6-Glucosidase Attacks α-1,6-glucose linkages of amylopectin
Source: Data from Garcia, E., and Lajola, A.M., Journal of Food Science 53,
1181–1186, 1988; Tucker, G.A., Biochemistry of Fruit Ripening,
­
1–52, 1993.

(Fait et al., 2008), and peach (Lo Bianco and Rieger, 2002) is evident dur-
ing ripening. The main sugar at the fruit ripening stage depends on the
plant species. Glucose is the major sugar in table grape, whereas fructose
is the predominant sugar in berries, mango, and citrus species. Those
fruits that accumulate fructose and glucose show very low concentrations
of sucrose. However, apricot, plum, nectarine, and peach have sucrose as
the main sugar, which is accumulated during stage 3 as a result of a rise
in the activity of sucrose synthase (Morandi et al., 2008). The proportions
of fructose, glucose, and sucrose are important in the perception of taste
since fructose is 80% sweeter than sucrose, whereas glucose is only 60%
sweeter than sucrose (Yamaguchi et al., 1970). A regular increase in the
level of sugars in pear throughout the period of fruit development has been
observed due to translocation of photosynthates from leaves to the young
fruit, which are partly used for the synthesis of pectic substances and other
cell wall materials and partly converted to the usual storage product, the
starch. With the advancement of maturity, the accumulated starch is hydro-
lyzed into sugars, which is known as a characteristic event for fruit ripening
(Hulme, 1958). The starch-degrading enzymes in fruits and their mode of
starch mobilization are given in Table 1.6. Further breakdown of sucrose
into glucose and fructose is probably mediated by the action of invertase.
Low levels of invertase activity have been linked to the sucrose accumula-
tion trait in tomatoes (Yelle et al., 1991).

1.3.3  Organic Acid Changes

The peculiar taste and flavor of fruit is dependent on the type of organic
acid, predominant organic acid, and ratio between the organic acid and
sugar. The predominant organic acid in ripe fruit varies among species.

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

As quoted by Etienne et al. (2013), “Malic acid is dominant in apple

(Yamaki, 1984), loquat (Chen et al. 2009) and pear (Lu et al. 2011), whereas
citric acid is dominant in citrus fruits (Yamaki, 1989). In many fruit spe-
cies, differences in total acidity or in the balance of organic acids among
cultivars are also observed, for example in loquat (Yang et al. 2011), peach
(Etienne et al. 2002), pear (Lu et al. 2011), citrus (Albertini et al. 2006),
pineapple (Saradhuldhat and Paull, 2007), apricot (Gurrieri et al. 2001) and
banana (Bugaud et al. 2011).” Loss of acidity implies decarboxylation of
carboxylates, which can occur through the conversion of tricarboxylates
into dicarboxylates, but also through decarboxylation of the dicarboxylates
malate and oxaloacetate (OA A), leading to the degradation of organic acids
(Figure 1.4). Decarboxylation of OA A and malate allows the production of

Pyruvate GLYOXYSOME
PDH
CoA
NAD-ME acetylCoA CoA
acetylCoA citrate citrate
CS ACO
OAA CS citrate OAA
OAA
ACO glyoxylate
NAD-MDH
cycle isocitrate isocitrate
malate NAD-MDH
cis-aconitate
malate malate ICL
ACO
TCA Cycle
fumarate MS
isocitrate glyoxylate
succinate
NAD-IDH CoA
succinate CoA NADP-IDH
2-oxoglutarate
succinyl-CoA
citrate ATP-CL acetylCoA
MITOCHONDRION OAA metabolism
GABA shunt
ACO acetylCoA
NADP-ME isocitrate
Pyruvate malate
NADP-IDH flavonoids/
2-oxoglutarate isoprenoids
PPDK NAD-MDH
gluconeogenesis
PEPCK glutamate
glucose PEP OAA
glutamine
PEPC GABA
glucolysis
CYTOSOL
succinate

Figure 1.4  Citrate and malate metabolic pathways in fruit mesocarp cells.
The probable direction of reversible reactions is indicated by the large
arrow. ACO, aconitase; ATP-CL, ATP-citrate lyase; CS, citrate synthase; ICL,
isocitrate lyase; MS, malate synthase; NAD-MDH, NAD-malate dehydro-
genase; NAD-ME, NAD-malic enzyme; NAD-IDH, NAD-isocitrate dehy-
drogenase; NADP-ME, NADP-malic enzyme; NADP-IDH, NADP-isocitrate
dehydrogenase; PDH, pyruvate dehydrogenase; PEPC, phosphoenolpyru-
vate carboxylase; PEPCK, phosphoenolpyruvate carboxykinase; PPDK, pyru-
vate orthophosphate dikinase. (Adapted from Etienne, A. et al., Journal of
Experimental Botany, doi:10.1093/jxb/ert0352013.)

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

phosphoenolpyruvate (PEP) and is linked to the activation of gluconeogen-

esis (Sweetman et al., 2009). Gluconeogenesis is a m ­ etabolic pathway that
results in the generation of glucose from PEP. It occurs mostly during fruit
ripening when sugars accumulate rapidly (Sweetman et al., 2009).
The downward trend in the levels of organic acids with the onset of
fruit ripening has been reported by a number of workers. The total organic
acid (malic + citric + quinic) decreased in peach with the ripening of fruits,
coinciding with decreasing titratable acidity (Wang et al., 1993). Being a
major respiratory substrate, organic acid (predominating) is metabolized
to a greater extent than the others and may fall by 50% during the life of the
fruit. Malic acid, as a major substrate of respiration, has been suggested,
and thus accounts for the respiratory quotient of 1.1 or higher, which is
typical of pome fruit (Fidler and North, 1967). The decline in the content
of organic acids during fruit ripening might be the result of an increase in
membrane permeability, which allows acids to be stored in the respiring
cells (Kliewer, 1971), as well as formation of salts of malic acid, reduction in
the amounts of acid translocated from the leaves, reduced ability of fruits
to synthesize organic acids with fruit maturity (Hardy, 1968), translocation
into sugars (Hulme, 1970), and dilution effect due to the increase in volume
of fruit.

1.3.4  Flavor and Aroma Changes

Flavor is the most important factor determining whether consumers will
repurchase a particular fruit. Two main factors determine a fruit’s char-
acteristic flavor: the correct sugar–acid balance and the production of
aroma volatile compounds. These volatile compounds can include a mix-
ture of volatile acids, aldehydes, alcohols, esters, terpenoids, and aromatics
(Table 1.7).
Flavor changes deal with the generation of volatiles to alter aroma,
the generation of sugars from stored starch, the interconversion of sugars,
a decline in acidity through altered organic acid metabolism, and a reduc-
tion in astringency through lower levels of tannins and phenolics (Kays,
1991). Even in a fruit as relatively nonaromatic as breadfruit, some 40
distinct volatile compounds are produced during ripening (Iwaoka et al.,
1994). Typically, only a handful of these volatiles, termed character impact
compounds, are the source of the distinctive odors associated with this
fruit. The second aspect of flavor relates to carbohydrate and organic acid
metabolism, and mango provides a good example. As it ripens, starch is
broken down, with glucose being converted to fructose and sucrose, while
at the same time various organic acids, chief among these succinate, are
depleted (Lazan et al., 1993). Some fruits have high levels of soluble phe-
nolics when immature. These result in astringency; during ripening, these

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.7  Aroma Substances in Fruits and Description of Their Odor

Aroma Substance Odor Description
(E)-2-Hexenal ‘Hayward’ kiwifruit
1-Methyl ethyl butyrate Apple
2-Hexenal Green leaf, green banana
2-Methyl butyl acetate Apple
2,6-Nonadienal Cucumber
Acetaldehyde Pungent, penetrating
Acetone Sweet, pungent
Allyl isothiocynate Raw cabbage
Butyl acetate Fruity
Butyl butyrate Fruity, pear
Citral Lemon
Dimethyl disulfide Cooked cabbage
Dimethyl disulfide Onion, cabbage
Ethyl acetate Etherlike, pineapple, anise
Ethyl butanoate ‘Hort16A’ kiwifruit
Ethyl butyrate Fruity, pineapple
Ethyl hexanoate Fruity
Eugenol Ripe banana
Furaneol Strawberry
Heptanone Banana
Hexanal Cut grass
Hexenal Sweet, almond, green
Hexyl acetate Fruity, apricot
Isopentanol Overripe banana
Linalool Fruity, floral, citrus
Methoxypyrazine Grassy
Methyl anthranilate Foxy
Methyl butyrate Apple
Methyl hexanoate Etherlike, pineapple
Monoterpene limonene Lime
Nootakatone Grapefruit
Raspberry ketone Raspberry
Valencene Orange
α-Farnesene Apple
β-Damascenone Richness
β-Lanon-trans Warm, woody, balsamic, rose

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

are polymerized, and astringency is lost, as these tannins are no longer

capable of interacting with taste receptors (Kays, 1991).
Watson et al. (2002) noted that many volatile and nonvolatile com-
pounds give rise to the flavor of strawberry fruit. The volatile compounds
give the fruit its distinctive flavor, whereas the nonvolatile compounds, such
as sugars and organic acids, are responsible for the fruit’s sweetness and
tartness. In tomato, several sensory attributes have been proposed to char-
acterize aroma, such as fruity, green, grassy, earthy, musty, floral, candy,
citrus, grapefruit, and pharmaceutical aromas (Causse et al., 2001; Baldwin
et al., 2004; Sinesio et al., 2010). More than 400 aroma volatiles have been
identified in tomato fruit, among which about 30 seem to be important for
tomato aroma (Baldwin et al., 2000, 2004). In apple and strawberry, more
than 300 volatile compounds have been identified (Dixon and Hewett,
2000), for which 20 volatiles are considered the aroma fingerprint of straw-
berry (Ulrich et al., 1997).
Amyl esters give bananas their distinctive flavor and aroma, and
butyl esters give them a fruity flavor and aroma; however, other esters and
aldehydes, alcohols, and ketones have been associated with flavor, and their
production rates can increase during ripening (Tressl and Jennings, 1972).
McCarthy et al. (1963) also claimed that amyl esters gave bananas their
distinctive flavor and aroma, and butyl esters gave them a fruity flavor and
aroma. Volatile compounds of several banana cultivars have been widely
studied by many authors: ‘FLHORBAN 920’ and ‘Grand Naine’ (Bugaud
et al., 2009); ‘Gran Enana’, a subgroup of the ‘Cavendish’ originating from
Central and South America (Vermeir et al., 2009); various cultivars grown
on Madeira Island (Nogueira et al., 2003); banana fruits (Musa sapientum L.
var. ‘Cavendish’) from Honduras and their aqueous essences (Jordan et al.,
2001); free and glycosidically bound volatile compounds of the ‘Valery’ and
‘Pequeña Enana’ cultivars (Pérez et al., 1997); banana fruits (Musa caven-
dishii L.) of the ‘Gran Enana’ and ‘Enana’ cultivars from the Canary Islands
and the ‘Enana’ cultivar from Colombia (Cano et al., 1997); Philippine
bananas (‘Del Monte’, ‘Cavendish’); Taiwanese bananas (cv. ‘Sen-nin’); and
‘Delicious’ bananas (hybrid between ‘Philippine’ and ‘Taiwanese’) (Shiota,
1993). Generally esters such as butyl acetate, isoamyl acetate, ethyl acetate,
butyl butanoate, and isoamyl isobutanoate are responsible for the charac-
teristic aroma of fresh banana and constitute the major class of compounds
present in banana’s volatile profile (Salmon et al., 1996). The typical aroma
of banana is characterized by the presence of a wide range of volatile metab-
olites—with different volatilities and concentrations that can vary among
the different cultivars—as the initial work of Cano and collaborators with
Spanish and Columbian ‘Enana’ cultivars showed (Cano et al., 1997). They
identified the compounds and the ratios of the peak area to the internal stan-
dard area corresponding to the gas chromatography–mass spectrometry
(GC-MS) of the purge-and-trap analysis of three banana cultivars (Spanish

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

‘Enana’, Spanish ‘Gran Enana’, Latin American ‘Enana’). Quantifiable

­differences among the flavors of the banana cultivars were found. Spanish
‘Enana’ fruit was found to be the richest in flavor compounds. McCarthy
et al. (1963) classified the various components of banana aroma. A banana-
like flavor was assigned to the amyl and isoamyl esters of acetic, propionic,
and butyric acid, whereas the alcohols and carbonyls gave odors described
as green, woody, or musty. In their study, the Latin American banana flavor
displayed the presence of ethanol, 1-butanol, and hexanal, but these com-
pounds were not found in any of the Spanish banana varieties. Furthermore,
only Spanish bananas exhibited the presence of hexyl butanoate, which is
related to a banana-like flavor.
Pontes et al. (2012) evaluated the effect of the cultivar (‘Dwarf
Cavendish’, ‘Prata’, ‘Maçã’, ‘Ouro’, and ‘Platano’) on the volatile profile
determined by dynamic headspace solid-phase microextraction (dHS-
SPME) combined with one-dimensional gas chromatography–mass spec-
trometry (1D-GC-MS). This approach allowed the definition of a volatile
metabolite profile to each banana variety and can be used as pertinent
criteria of differentiation. A total of 68 volatile organic metabolites were
tentatively identified and used to profile the volatile composition in differ-
ent banana cultivars, thus emphasizing the sensitivity and applicability of
SPME for establishment of the volatile metabolomic pattern of plant sec-
ondary metabolites. Ethyl esters were found to comprise the largest chemi-
cal class, accounting for 80.9%, 86.5%, 51.2%, 90.1%, and 6.1% of total peak
area for ‘Dwarf Cavendish’, ‘Prata’, ‘Ouro’, ‘Maçã’, and ‘Platano’ volatile frac-
tions, respectively. de Vasconcelos Facundo et al. (2012) determined the
volatile differences in two cultivars of banana under cold storage conditions.
Cold storage more strongly affects the ‘Nanicão’ than the ‘Prata’ cultivar.
Esters such as 2-pentanol acetate, 3-methyl-1-butanol acetate, 2-methylpro-
pyl butanoate, 3-methyl butyl butanoate, 2-methylpropyl 3-methyl butano-
ate, and butyl butanoate were drastically reduced in the cold group of the
‘Nanicão’ cultivar.
A comparative study of volatile components of 9 litchi cultivars (10
samples) from southern China was carried out through GC-MS com-
bined with headspace SPME (Wu et al., 2009). A total of 69 volatiles were
detected, of which 43 were identified and another 53 were tentatively
identified: 35  terpenoids, 27 alcohols, 10 aromatic compounds, 9 alde-
hydes, 8  esters, 4  ketones, 2 sulfurs, and 1 organic acid. Seventeen com-
mon volatiles included in all the samples were linalool, cis-rose oxide,
R-terpineol, β-citronellol, geraniol, p-cymene, ethanol, 3-methyl-3-buten-
1-ol, 3-methyl-2-buten-1-ol, 1-hexanol, (E)-2-hexen-1-ol, 2-ethyl-1-hexanol,
1-octen-3-ol, 1-octanol, ethyl acetate, p,R-dimethylstyrene, and 3-tert-butyl-
4-hydroxyanisole. In these cultivars, ‘Guangxi Huaizhi’ contained at most
67 volatiles, and ‘Jizhuili’ contained at least 36 volatiles. Alcohols were the
predominant volatile components in various cultivars, representing 35.1%

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

(‘Guangdong Huaizhi’) to 81.6% (‘Jizhuili’) of the main fraction. Although

the volatile composition and concentration varied between these cultivars,
the components with the highest odor activity values (OAVs) in most culti-
vars were still cis-rose oxide, trans-rose oxide, 1-octen-3-ol, and geraniol.
Two Huaizhi samples from two producing areas exhibited similar volatile
profiles and were significantly different from other cultivars according to
cluster analysis performed on amounts of major volatile components (Wu
et  al., 2009). Fourteen volatile compounds, namely, three alcohols, one
ester, three monoterpenes, and seven sesquiterpenes, were detected at har-
vest in cultivar Mauritius in South Africa. Cultivar McLean’s Red showed
19 volatile compounds, namely, 4 alcohols, 3 monoterpenes, 2 oxides, and
10 sesquiterpenes. In cultivar Mauritius, alcohols represented 50% of the
main fraction, followed by sesquiterpenes (23%), monoterpenes (21%), an
unknown component (6%), and the ester (0.7%). No ester was detected in
cultivar McLean’s Red (Sivakumar et al., 2008). Among the three alco-
hols, citrinelol and geraniol predominated in the aroma profile of cultivar
Mauritius, conferring a characteristic “floral, rose, citrus and fruity aroma”
to the fruit (Chyau et al., 2003). Limonene, rose oxide, citronellol, and gera-
niol were detected at relatively low levels in cultivar Mauritius, although
rose oxide was not present in the latter. Zingiberene was the predominant
sesquiterpene, and terpinolene the most abundant monoterpene in cultivar
Mauritius (Sivakumar et al., 2008).

1.3.5  Cell Wall and Textural Changes

Recently several reviews (Vicente et al., 2007d; Goulao and Oliveira, 2008;
Li et al., 2010; Cruz-Hernandez and Paredes-Lopez, 2012) have been pub-
lished on the role of cell wall–degrading enzymes and textural changes.
Fruit softening and related textural alterations during ripening are mainly
consequences of progressive depolymerization and solubilization of cell wall
components and loss of cell structure. Both enzymatic and nonenzymatic
factors contribute to softening (Brummell and Harpster, 2001; Dumville
and Fry, 2003; Brummell, 2006). Moreover, different fruits soften at dif-
ferent rates and by varying degrees due to the inherent composition and
nature of their cell wall polysaccharides and other cell wall structural com-
ponents (Table 1.8) (Tucker and Grierson, 1987; Brummell, 2006). Fruit
such as banana, mango, kiwifruit, avocado, and papaya undergo dramatic
softening; fruit such as apple, grape, and citrus do not exhibit such drastic
changes. Different rates of softening can also be illustrated by varied dura-
tions required for loss of firmness for different fruit types; for example,
comparative studies showed that fruit softening at room temperature may
take 3 days for both tomato and banana, 4.5 days for mango, 30 days for
carambola, and 24 days for ‘Kampuchea’ guava (Ali et al., 2004).

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.8  Common Polymers of Primary Cell Wall and Their Structures
Polymer Molecular Structure
Cellulose (1→4)β-D-Glucan chains held together with
hydrogen bonding, forming very long crystalline
microfibrils
Cross section contains 36 glucan chains
Glucomannan Backbone contains regions of (1→4)β-D-glucan and
(1→4)β-D-mannan in nearly similar amounts
galactomannan
Occasionally terminal; has a side chain of single
units of α-D-galactose
Glucuronoarabinoxylan Backbone of (1→4)β-D-xylan
Side chains of single unit of nonreducing terminal
α-L-arabinose and α-D-glucoronic acid
Homogalacturonan Made of long chains of (1→4)α-D-galacturonic acid
Initially highly methyl esterified
Rhamnogalacturonan I Made of alternating α-D-rhamnose and α-D-
(RG-I) galacturonic acid residues; long side chains of
either unbranched (1→4)β-D-galactan or branched
α-L-arabinans or type I arabinogalactans attached
to the rhamnose residues
Rhamnogalacturonan II Backbone made of (1→4)α-D-galacturonic acid like
(RG-II) homogalacturohan; complex side chains of
different types of neutral sugar
A minor cell wall component
RG-II monomers can domimerize together as boron
diesters and may affect the cell wall porosity
Structural proteins Four different types including expansin; some are
heavily glycosylated
Xyloglucan Backbone similar to cellulose, i.e., (1→4)β-D-glucan
Regular substitution on three out of four consecutive
glucose residue with α-D-xylose
Xylose occasionally extended with β-D-galactosyl-α-L-
fucose or α-L-arabinose in some species
The reducing end of unsubstituted glucose residues
is susceptible to cleavage by Trichoderma endo-
(1→4)β-D-glucanase (EGases) producing similar
amounts of heptasaccharide (G1c4.Xy13) and
nonasaccharide (G1c4.Xy13.Gal.Fuc) xyloglucan
subunit oligosaccharides

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Fruit cell walls consists of complex networks of polysaccharides and

proteins, wherein the primary cell wall contains, on average, about 35% pec-
tin, 25% cellulose, 20% hemicellulose, and 10% structural protein, depending
on the fruit species and developmental stage (Brownleader et al., 1999).
Major classes of cell wall polysaccharides are modified to varying levels
during fruit ripening, and a scheme depicting softening and those cell wall
modifications occurring in ripening has been proposed for a melting-flesh
peach by Toivonen and Brummell (2008). In general, cell wall modification
and depolymerization during fruit ripening are variable among different
fruits (Brummell, 2006; Toivonen and Brummell, 2008). For example, pec-
tin depolymerization has been reported as the major cause of loss of firm-
ness in raspberry and boysenberry (Vicente et al., 2007a, 2007c), whereas
only slight pectin depolymerization is detected in ripening strawberry
(Huber, 1984), banana (Wade et al., 1992), and blueberry (Vicente et al.,
2007b). Major cell wall enzymes assayed in different fruits during ripening
are given in Table 1.9.

Table 1.9  Cell Wall Enzymes Assayed in Fruit Ripening

Fruits Enzymes
Climacteric Fruits
Apple β-Galactosidase, glucanase, polygalacturonase, pectin
methyl esterase (PME)
Avocado Glucanase, polygalacturonase
Banana Glucanase, polygalacturonase, PME
Kiwi Polygalacturonase, xylanase (xyloglucan
endotransglicoylase)
Mango β-Galactosidase, PME
Melon β-Galactosidase, glucanase, polygalacturonase, PME
Papaya β-Galactosidase, glucanase, polygalacturonase, PME
Peach β-Galactosidase, glucanase, polygalacturonase, PME
Pear β-Galactosidase, β-glucosidase, polygalacturonase
Pepper Glucanase
Tomato Glucanase, polygalacturonase, PME, xilanase
Nonclimacteric Fruits
Grape α-Galactosidase, β-galactosidase, PME
Orange α-Galactosidase, β-galactosidase, glucanase,
α-glucosidase, β-glucosidase, PME
Prickly pear β-Galactosidase, glucanase, polygalacturonase, PME
Strawberry Glucanase, PME

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Softening in kiwifruit occurs over a period of weeks and can be

divided into a number of phases (Figure 1.5). Modifications of the cell wall
play an important part in determining fruit texture and ripening charac-
teristics. Chemical analyses of cell wall components show some consistent
changes during the early stages of ripening. These include

1. Solubilization of pectin (but without further degradation)

2. The cell wall swelling and showing an increased affinity for water
(becoming more hydrophilic)
3. Loss of galactose from pectins (especially of a galactan that is
tightly associated with the cellulose microfibrils)
4. Deesterification of some pectins

These changes continue once kiwifruit have begun rapidly softening to

ripeness (phase 2 in Figure 1.5). Phase 2 softening is associated with a
further increase in pectin solubilization, loss of galactan and arabinan side
chains from pectic polymers, and more cell wall swelling. As softening pro-
gresses into phase 3, two more important changes begin, both of which
appear to be regulated by ethylene:

5. Depolymerization (a reduction in size) of the hemicellulosic poly-

saccharide xyloglucan, which is associated with a reduction in cell
wall strength
6. Depolymerization of pectin, which is associated with dissolution of
the middle lamella and reduced intercellular adhesion.

These six changes have been observed in a wide range of fruit types,
although the extent and relative timing vary somewhat between species.
Such observations indicate that pectin solubilization and cell wall swelling
are important events in the control of softening in kiwifruit and probably
most other species with melting texture.
Graham Seymour and colleagues at HRI–Wellesbourne are investi-
gating the biochemistry of fruit texture in order to improve the palatabil-
ity and shelf life of produce (Marin-Rodriguez et al., 2002; Seymour et al.,
2002). Marin-Rodriguez et al. (2002) reviewed the role of pectate lyases in
fruit softening. Pectate lyases (PELs) catalyze the Ca 2+ -dependent cleavage
of deesterified pectin, which is a major component in the primary cell walls
of many higher plants. Initially, it was thought that these enzymes were
produced solely by plant pathogens to macerate plant tissues. As a result of
both plant genome sequencing and EST programs, however, it has become
clear that these enzymes are encoded by large gene families in plants and
expressed throughout the plant, including ripening fruit. Tomato, straw-
berry, grape, and banana fruits all express PELs, where they may play a
significant role in fruit softening. Other enzymes involved in modifying

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Respond to exogenous ethylene Autocatalytic ethylene production

100
Starch degradation

80
Pectin solubilization
Fruit firmness (%)

60 Phase 1
initiation
Soluble pectin depolymerization
galactose loss
40

Aroma production
respiratory climacteric
Phase 2 pectin depolymerization
20 rapid softening
Loss of middle lamella
Phase 3
eating window Phase 4
over-ripe
Time

Figure 1.5  Schematic representation of postharvest ripening in kiwifruit,

showing the timing of key physiological events. At harvest, fruit do not pro-
duce ethylene but are highly sensitive to exogenous ethylene. Softening is
initiated (phase 1) and becomes rapid (phase 2). Relatively late in softening,
compared with other fruit species, endogenous autocatalytic ethylene pro-
duction begins, aroma volatiles are produced, and fruit become soft enough
to eat (phase 3). If fruit progress to the overripe stage (phase 4), they become
unacceptably soft and exhibit off-flavor notes. (Reproduced from Atkinson
R.G. et al., Journal of Experimental Botany 62, 3821–3835, 2011.)

cell wall properties include pectin esterases (PEs) and polygalacturonases

(PGs). These enzymatic activities (Table 1.10) are similarly encoded by
multigene families, in which at least one member shows ripening-specific
expression.
Several studies have been carried out to investigate the role of cell
wall degradation-related enzymes and the expression of respective genes
during softening of the grape berries (Nunan et al., 2001; Ishimaru and
Kobayashi, 2002; Waters et al., 2005). Chervin et al. (2008) showed that
low doses of ethylene application increased the berry diameter at the incep-
tion of the ripening stage. The study verified previous studies that showed
correlation between berry ripening and the accumulation of various

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.10  Pectin-Modifying and -Degrading Enzymes in Fruits

Substrate Enzymes Products
Pectin Pectin methyl esterase Pectic acid + methanol
Endopolymethyl Methyl oligogalacturonides
galacturonase α-(1,2)-linked L-Rha-α-(1,4)-
linked D-Gal
Endopectinlyases Unsaturated
oligogalacturonides
Hairy pectin Rhamnogalacturonan Pectin + acetic acid
acetyl esterase
Pectin acetyl esterase Pectin + acetic acid
Smooth pectins Lyases Oligogalacturonides
Protopectin Protopectinase Pectin
Pectic acid Endo-PG Oligogalacturonides
Exo-PG Monogalacturonides
Endopectate lyases Oligogalacturonides
Exopectate lyases Unsaturated
digalactoronides
Trigalacturonic Oligogalacturonide Monogalacturonides
acid hydrolase
∆4:5 ∆4:5 unsaturated Unsaturated
(galacturonide) n oligogalacturonide monogalacturonide +
hydrolase galacturonides (n − 1)
Unsaturated Oligogalacturonide lyases Unsaturated
digalacturonate monogalacturonides
Arabinans α-L-Arabinofuranosidase α-L-Arabinose
(1,5)-α-Arabinans Endoarabinanase Arabinose and higher
oligosaccharides
Galactans β-D-Galactanase β-D-Galactose

transcripts of PG, PME, cellulose, and expansion. Some of the enzymes

may be involved in the increase of berry expansion by ethylene.
Manenoi et al. (2007) determined endixylanase gene expression, pro-
tein amount, and activity in three papaya cultivars that differ in softening
pattern and in one cultivar where softening was modified by the ethylene
receptor inhibitor 1-MCP. Differential expression of gene and enzyme activ-
ity was noticed in different cultivars and one treated with 1-MCP. The xyla-
nase gene could be utilized to control the growth and abscission.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

1.3.6  Physiological Changes

On-tree and off-tree ripening physiology is dependent on respiration and eth-
ylene responses and concentration in fruit. As discussed earlier, fruits can be
classified into climacteric and nonclimacteric categories, depending on respi-
ration peaks and response to exogenous ethylene. Therefore, it is important to
study the respiration and ethylene behavior during ripening and postharvest
in fruits. Here respiration rates, ethylene concentrations, and their changes
during growth and ripening in various fruits are discussed (Table 1.11). Other
aspects related to physiological changes during on-tree ripening and posthar-
vest ripening are discussed throughout the book in various chapters.

Table 1.11  Respiration and Ethylene Production Rates Measured at 20°C in

Several Climacteric and Nonclimacteric Fruits
Respiration Rates Ethylene Production
Fruit (mg CO2 kg−1 h−1) (µlC2H4 kg−1 h−1)
Apple 20–31 25–2500
Apricot 40 < 0.01 (0°C)
Araza 1283 nd
Asian pear 25
Atemoya 250 200 (20°C)
Avocado 190 28.9–74.2
Banana 280 0.05–2.1
Blackberry 115 0.1–2.0
Blueberry 70 0.5–10.0
Breadfruit 480 (25°C) 1.2
Carambola 65 < 3.0
Cherimoya 300 200
Cherry, sweet 65 < 0.01 (0°C)
Cranberry 16 0.6 (5°C)
Dragon fruit 105 < 0.1
Durian 265 40
Fig 50 0.6 (0°C)
Gooseberry 81 nd
Grape, American 33 < 0.1
Grape, muscadine 51 < 0.1
Grape, table 27 < 0.1
Grapefruit < 10 (15°C) < 0.1

(Continued )

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Table 1.11  Respiration and Ethylene Production Rates Measured at 20°C in

Several Climacteric and Nonclimacteric Fruits
Respiration Rates Ethylene Production
Fruit (mg CO2 kg−1 h−1) (µlC2H4 kg−1 h−1)
Guava 74 10
Honeydew melon 30 Very low
Kiwifruit 19 75
Lemon 24 0.11–0.17
Lime nd 0.30–1.96
Litchi 60 Very low
Longan 42 Very low
Loquat 80 Very low
Mamey apple 35 (25°C) 400.0 (27°C)
Mandarin 25 < 0.1
Mango 58 0.04–3.0
Mangosteen 21 (25°C) 0.03
Nectarine 87 5.0 (0°C)
Netted melon 55.0 55.0
Orange 28 < 0.1
Papaya 80 8.0
Passion fruit 262 280.0
Peach 87 0.9–20.7
Pepper 34 < 0.2
Persimmon 22 < 0.5
Pineapple 24 0.16–0.40
Plum 20 0.14–0.23
Pomegranate 24 < 0.1
Prickly pear 32 0.2
Rambutan nd Very low
Raspberry 125 12.0
Sapodilla 16 (25°C) 3.7
Sapote nd >100
Star apple 38 0.1
Strawberry 150 < 0.1
Tomato 35 3.6–29.8
Watermelon 21 < 1.0
Wax apple 10 Very little
Note: nd, Not detected.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

The respiration rate of avocado fruit is relatively high compared

to that of many other fruits: about 20–50 mg CO2 kg−1 h−1 at 5°C, 50–160
mg CO2 kg−1 h−1 at 10°C, and 80–300 mg CO2 kg−1 h−1 at 20°C (Kader and
Arpaia, 2001). Rates of ethylene production are generally low for unripe
avocados, < 0.1 µl kg−1 h−1 at 20°C, but increase rapidly after harvest up to
> 100 µl kg−1 h−1 at 20°C when fully ripe. Thus, the implications for ethylene
in avocados depend on the physiological state of the fruit.
Carambola exhibits nonclimacteric ripening behavior. Carambola
respiration was fairly constant at 20°C, < 20 ml CO2 kg−1 h−1 at 20°C
(Warren, 2009). It increased slightly while turning from yellow to orange,
likely owing to senescence of the fruit. Ethylene production was very low
at 25°C, about 0.4 µl kg−1 h−1. In an another study, fruit at the full-ripe (one-
quarter orange) stage had higher respiration and ethylene production rates
(Oslund and Davenport, 1983).
The respiration rate of dates is very low, < 5 mg CO2 kg−1 h−1 at 20°C
at the khalal stage and < 1 mg CO2 kg−1 h−1 at the rutab and tamr stages
(Yahia, 2004). Cured ‘Deglet Noor’ dates with 20%–22% moisture produced
0.4 mg CO2 kg−1 h−1 at 24°C and 2 mg CO2 kg−1 h−1 when the moisture con-
tent increased to 27% (Rygg, 1975). The rate of CO2 production is high ini-
tially, but declines steadily as the fruit advances in maturity, reaching its
lowest level as the fruit enters the stage of physiological maturity and then
increases to reach a peak as fruit ripens (Abbas and Ibrahim, 1996). Dates
produce very low concentrations of ethylene: < 0.5 µl kg−1 h−1 for the kha-
lal stage and < 0.1 µl kg−1 h−1 for the rutab and tamr stages kept at 20°C
(Yahia, 2004). Ethylene production in ‘Hallawi’ dates was not detected until
91 days after pollination; it increased to reach a peak within 15 days and
then declined rapidly (Abbas and Ibrahim, 1996).
Durian has a climacteric respiration characteristic with a peak rate
of around 200–250 mg CO2 kg−1 h−1 at 25°C; ethylene production ranges
between 0.3 and 1.4 µl C2H4 kg−1 h−1 during the preclimacteric stage and 8
and 12 µl C2H4 kg−1h−1 at its peak (Siriphanich et al., 1994). Internal atmo-
spheric composition changes with the ripening stage. CO2 increased from
1%–4% at harvest to 4%–14% when ripened at 22°C, while ethylene increases
from 0.5–1 µl L −1 to 3–7 µl L −1 (Tongdee et al., 1990).
Grape is a nonclimacteric fruit with a relatively low respiration rate
(3–4 ml CO2 kg−1 h−1 at 5°C). At 5°C, grapes produce 50% less heat of respi-
ration than other nonclimacteric fruits. Comparison of the cluster compo-
nents indicates that the respiration rate of the rachis alone is approximately
15 times higher than that of the berry at 4°C (Gardea et al., 1994). Grapes
produce very small amounts of ethylene during berry development. There
is peak of ethylene production at bloom, followed by a decreasing concen-
tration until harvest (Weaver and Singh, 1978).
During guava fruit ripening, ethylene production and respiration
rates of fruit harvested at the full-size, pale-green stage were higher than

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 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

those of fruit harvested at the small- or medium-size, dark-green stage

(Brown and Wills, 1983). Fruit harvested at advanced maturity reach their
respiratory climacteric stage in 4–6 days, accompanied by rapid changes
in skin color and flesh firmness. Ethylene production rates increase dur-
ing fruit ripening and reach a peak that may or may not coincide with the
respiratory peak. Guava cultivars such as ‘Allahabad Safeda’, ‘Apple Color’,
and ‘Hisar Safeda’ produce higher amounts of ethylene than ‘Lucknow-49’
(Singh and Pal, 2008).
Jackfruit respiration behavior follows a typical climacteric pattern
with three distinct phases of respiration (preclimacteric, climacteric, and
postclimacteric stages). Precut immature fruit slices showed a basal respi-
ration rate of 160 mg CO2 kg−1 h−1 fruit, and in the climacteric phase, the
peak of respiration was 245 mg CO2 kg−1 h−1 fruit (Selvaraj and Pal, 1989).
The climacteric peak was attained after 3 days of ripening. After 8 days of
ripening, the respiration rate had dropped to a residual level of 60–70 mg
CO2 kg−1 h−1 fruit.
Climacteric behavior in Chinese jujube is cultivar dependent. Some
of the cultivars show a climacteric nature of ripening, while few have
a nonclimacteric ripening pattern. Shen et al. (2004) reported that the
respiration rate and ethylene production of ‘Dongzao’ jujube at 4°C and
20°C each exhibited increasing tendencies with extension of storage time,
but that no apparent peaks of respiration or ethylene production were
detected. This suggested that ‘Dongzao’ jujube fruit is nonclimacteric.
In contrast, Jiang and Sheng (2003) found that the rates of respiration
and ethylene production of ‘Jins’ jujube during storage initially increased
and then reached respective peaks before finally decreasing rapidly; they
were further promoted when the fruit was exposed to ethylene. These
observations suggest that ‘Jins’ cultivar is climacteric. Indian jujube has a
high respiration rate and a climacteric respiration pattern (Pareek et al.,
2009; Pareek and Yahia, 2013). The rate of respiration of mature green
Indian jujube fruit of both Ziziphus mauritiana and Ziziphus spina-christi
is low but increases as the fruit matures, reaching a peak value when it
enters the ripening phase and then declining rapidly (Abbas, 1997). Abbas
and Fandi (2002) found that the respiration rate was high 3 weeks after
anthesis, but then declined steadily until 12 weeks after anthesis, when it
reached 14.2 mg CO2 kg−1 h−1. The rate then began to increase, first slowly
and then rapidly to a maximum value of 100.3 mg CO2 kg−1 h−1 as the fruit
entered the ripening phase. The rate of CO2 production declined rapidly
as the fruit became overripe. This pattern of respiratory change is charac-
teristic of a climacteric fruit.
Litchi fruit is nonclimacteric with relatively low levels of ethylene
production after harvest. The fruit does not ripen after harvest, and eth-
ylene production remains constant at 1°C–3°C storage temperatures for
30 days (Chen et al., 1968). According to Jiang et al. (1986), a decline in

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

the rate of respiration was observed during fruit development. In cultivar

‘Huaizi’, the respiration decreased directly after harvest, and thereafter
an increase was observed.
There is a controversial assessment concerning ethylene production
versus fruit maturity in loquat. Some authors found ethylene production
and an increase of respiration rate prior to color break (Gariglio et al., 2002;
Amoros et al., 2003), permitting Amoros et al. (2003) to define loquat as
a climacteric fruit. Hirai (1980) observed a marked increase in respira-
tion during fruit maturation, and the observation may have contributed to
the reclassification of loquat as a climacteric fruit by some authors, which
makes the classification of loquat controversial (Hasegawa et al., 2010).
However, others reported that although there is a well-defined peak of
ethylene production, it is insufficient to define loquat as a climacteric fruit
because there is not a concomitant increase in respiration rate, which often
appears ­irregular (Hamauzu et al., 1999; Ding et al., 1998; Kader, 2002;
Gonzalez et al., 2004). Therefore, there is no conclusive evidence to define
loquat fruit as a climacteric fruit, since (1) there is no starch accumulation at
stage 2 of fruit development (Gariglio et al., 2008); (2) there is no CO2 –C2H4
production relationship (Reig et al., 2007); (3) there is no general response
to e­ thephon to promote fruit ripening (Reig et al., 2007); (4)  there is no
confirmed evidence of an increase in hemicellulase, cellulose, and pectin
disrupting enzyme activities linked to ethylene production; and (5) there
is evidence of a nonclimacteric fruit postharvest behavior (Kader, 2002).
It is a well-established fact that mango exhibited a climacteric
pattern of respiration and an increase in ethylene production during
­r ipening. Respiration is very high after fruit set and then declines and
is maintained at a low rate until fruit ripening begins (Brecht and Yahia,
2009). Respiration patterns and ripening behavior vary among cultivars,
with different climatic conditions and growing locations. The respira-
tory peak in ‘Alphonso’ mangoes harvested when mature green occurs
5 days after harvest, and the fruit ripen within 7 or 8 days. ‘Piari’ man-
goes ripen on day 9 (Krishnamurthy and Subramanyam, 1970). The rise
in climacteric respiration in ‘Dashehari’, ‘Amrapali’, and ‘Rataul’ mangoes
coincides with the highest level of sucrose and PG activity in ripening
fruit (Kalra and Tandon, 1983). Mango fruit ripening is accompanied by
increased ethylene production, which coordinates the ripening process.
Mango expresses an autocatalytic increase in ethylene production during
ripening (Mattoo and Modi, 1969). Ethylene production starts before full
ripeness is reached (Cua and Lizada, 1990). Ethylene production in unripe
mango fruit is very low < 0.1 μl kg−1 h−1) (Burdon et al., 1996). Ethylene
production decreases as the fruit matures, is then undetectable for a
time, and reappears upon initiation of ripening (Akamine and Goo, 1979).
‘Kent’ and ‘Haden’ mango fruit have internal ethylene concentrations of
0.00  μl L −1 during the preclimacteric phase, increasing to 0.08 μl L −1 at

32
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

the initiation of the climacteric stage and up to 3.0 μl L −1 at the c­ limacteric

peak. Burg and Burg (1962) reported that ethylene production rises when
or before CO2 production rises in ripening mangoes, whereas Biale and
Young (1981) included mangoes among fruits in which ethylene rises
after CO2 production rises.
In papaya, the peaks in rates of respiration and ethylene production
coincide with full skin color development (Manenoi et al., 2007). Rates of
respiration and ethylene production began to rise after 2 days at 22°C in
‘Rainbow’ papayas were harvested at the color-break stage and reached
their maximum after 10 and 8 days of harvesting, respectively (Manenoi
et al., 2007). The number of days to reach respiratory and ethylene climac-
teric peaks and fruit ripening during postharvest also decreases with the
advancement of fruit maturity at harvest. For example, Golden’ papayas
harvested at different stages, viz. mature green, up to 15% yellow, up to 25%
yellow, and up to 50% yellow, reached the edible-ripe stage after 7, 6, 4, and
3 days at 23°C, respectively (Bron and Jacomino, 2006).

1.4  Conclusions and Future Perspectives

Remarkable progress has been made in understanding fruit ripening and
associated changes. Fruit flavor, aroma, softening, color, and sugar–acid
blend are important qualitative characteristics that determine the purchas-
ing of ripened fruit. Delaying ripening and maintaining quality throughout
the supply chain is the main objective of fruit physiologists and postharvest
technologists. Omics technology (proteomics, transcriptomics, genomics,
metabolomics) can be applied in this area. It has already provided a large
amount of information on metabolomic, proteomic, and transcriptomic
changes occurring during fruit ripening. However, very little has been
studied on systems biology for unraveling regulatory networks during fruit
ripening. A systems biology approach is needed for understanding climac-
teric and nonclimacteric fruit ripening. Epigenetics is another novel area
for fruit ripening study in the modern era.

References
Abbas, M.F. 1997. Jujube. In Mitra, S.K. (ed.), Postharvest Physiology
and Storage of Tropical and Subtropical Fruits. CAB International,
Wallinhgford, Oxfordshire, UK. pp. 405–415.
Abbas, M.F., and Fandi, B.S. 2002. Respiration rate, ethylene production
and biochemical changes during fruit development and maturation of
jujube (Ziziphus mauritiana Lamk). Journal of the Science of Food and
Agriculture 82, 472–476.

33
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Abbas, M.F., and Ibrahim, M.A. 1996. The role of ethylene in the regulation
of fruit ripening in the Hillawi date palm (Phoenix dactylifera). Journal of
the Science of Food and Agriculture 72, 306–308.
Aharoni, A., and O’Connell, A.P. 2002. Gene expression analysis of strawberry
achene and receptacle maturation using DNA microarrays. Journal of
Experimental Botany 53, 2073–2087.
Aina, J.O., and Oladunjoye, O.O. 1993. Respiration, pectolytic activity and tex-
tural changes in ripening African mango (Irvingia gabonensis) fruits.
Journal of the Science of Food and Agriculture 63, 451–454.
Akamine, E.T., and Goo, T. 1978. Respiration and ethylene production in mam-
mee apple (Mammea americana L.). Journal of the American Society for
Horticultural Science 103, 308–310.
Akamine, E.T., and Goo, T. 1979. Respiration and ethylene production in fruits
of species and cultivars of Psidium and species of Eugenia. Journal of the
American Society for Horticultural Science 104, 632–635.
Albertini, M.V., Carcouet, E., Pailly, O., Gambotti, C., Luro, F., and Berti, L.
2006. Changes in organic acids and sugars during early stages of devel-
opment of acidic and acidless citrus fruit. Journal of Agricultural and
Food Chemistry 54, 8335–8339.
Alexander, L., and Grierson, D. 2002. Ethylene biosynthesis and action in
tomato: A model for climacteric fruit ripening. Journal of Experimental
Botany 53, 2039–2055.
Ali, Z.M., Chin, L.H., and Lazan, H.A. 2004. A comparative study on wall
degrading enzymes, pectin modifications and softening during ripening
of selected tropical fruits. Plant Science 167, 317–327.
Amoros, A., Zapata, P., Pretel, M.T., Botella, M.A., and Serrano, M. 2003.
Physico-chemical and physiological changes during fruit development
and ripening of five loquat (Eriobotrya japonica Lindl.) cultivars. Food
Science and Technology International 9, 43–51.
Atkinson, R.G., Gunaseelan, K., Wang, M.Y., Luo, L., Wang, T., Norling,
C.L., Johnston, S.L., Maddumage, R., Schroder, R., and Schaffer, R.J.
2011. Dissecting the role of climacteric ethylene in kiwifruit (Actinidia
­chinensis) ripening using a 1-aminocyclopropane-1-carboxylic acid
­oxidase knockdown line. Journal of Experimental Botany, 62, 3821–3835.
Baldwin, E.A., Goodner, K., Plotto, A., Pritcett, K., and Einstein, M. 2004.
Effect of volatiles and their concentration on perception of tomato
descriptors. Journal of Food Science 69, 310–318.
Baldwin, E.A., Scott, J.W., Shewmaker, C.K., and Schuw, W. 2000. Flavor trivia
and tomato aroma: Biochemistry and possible mechanisms for control
of important aroma components. HortScience 35, 1013–1022.
Bapat, V.A., Trivedi, P.K., Ghosh, A., Sane, V.A., Ganapathi, T.R., and Nath, P.
2010. Ripening of fleshy fruit: Molecular insight and the role of ethyl-
ene. Biotechnology Advances 28, 94–107.
Ben-Arie, R., Segal, N., and Guelfat-Reich, S. 1984. The maturation and ripen-
ing of the ‘Wonderful’ pomegranate. Journal of the American Society for
Horticultural Science 109, 898–902.

34
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Biale, J.B. 1960. Respiration of fruit. In Ruhland, W. (ed.), Encyclopedia of

Plant Physiology, vol. 12. Springer, Berlin, pp. 536–592.
Biale, J.B. 1976. Recent advances in postharvest physiology of tropical and
subtropical fruits. Acta Horticulturae 57, 179–187.
Biale, J.B., and Barcus, D.E. 1970. Respiratory patterns of tropical fruits of
the Amazon basin. Tropical Science 12, 93–104.
Biale, J.B., and Young, R.E. 1981. Respiration and ripening in fruits—
retrospect and prospect. In Friend, J., and Rhodes, M.J.C. (eds.), Recent
Advances in the Biochemistry of Fruit and Vegetables, 1st ed. Academic
Press, London, pp. 1–39.
Blanpied, G.D. 1972. A study of ethylene in apple, red raspberry and cherry.
Plant Physiology 48, 627–630.
Bouzayen, M., Latche, A., Nath, P., and Pech, J.C. 2010. Mechanism of fruit
ripening. In Pua, E.C., and Davey, M.R. (eds.), Plant Developmental
Biology—Biotechnological Perspectives, vol. 1. Springer-Verlag, Berlin,
pp. 319–339.
Brady, C.J. 1987. Fruit ripening. Annual Review of Plant Physiology and Plant
Molecular Biology 38, 155–178.
Brecht, J., and Yahia, E.M. 2009. Postharvest physiology. In Litz, R. (ed.),
The Mango: Botany, Production and Uses, 2nd ed. CAB International,
Wallingford, UK, pp. 484–528.
Bron, I.U., and Jacomino, A.P. 2006. Ripening and quality of ‘Golden’ papaya
fruit harvested at different maturity stages. Brazilian Journal of Plant
Physiology 18, 389–396.
Brown, A.C., and Summers, W.L. 1985. Carbohydrates accumulation and
color development in watermelon. Journal of the American Society for
Horticulture Science 110, 683–686.
Brown, B.I., and Wills, R.B.H. 1983. Postharvest changes in guava fruits of
different maturity. Scientia Horticulturae 19, 237–243.
Brown, B.I., Wong, I.S., George, A.P., and Nissen, R.J. 1988. Comparative
studies on the postharvest physiology of fruit from different spe-
cies of Annona (custard apple). Journal of Horticultural Science 63,
521–528.
Brownleader, M.D., Jackson, P., Mobasheri, A., Pantelides, A.T., Sumar, S.,
Trevan, M., and Dey, P.M. 1999. Molecular aspects of cell wall modi-
fications during fruit ripening. Critical Reviews in Food Science and
Nutrition 39, 149–164.
Brummell, D.A. 2006. Cell wall disassembly in ripening fruit. Functional
Plant Biology 33, 103–119.
Brummell, D.A., and Harpster, M.H. 2001. Cell wall metabolism in fruit
softening and quality and its manipulation in transgenic plants. Plant
Molecular Biology 47, 311–340.
Bugaud, C., Alter, P., Daribo, M.O., and Brillouet, J.M. 2009. Comparison of
the physico-chemical characteristics of a new triploid banana hybrid,
FLHORBAN 920, and the Cavendish variety. Journal of the Science of
Food and Agriculture 89, 407–413.

35
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Bugaud, C., Deverge, E., Daribo, M.O., Ribeyre, F., Fils-Lycaon, B., and
Mbéguié-A-Mbéguié, D. 2011. Sensory characterisation enabled the
first classification of dessert bananas. Journal of the Science of Food and
Agriculture 91, 992–1000.
Burdon, J., Dori, S., Marinansky, R., and Pesis, E. 1996. Acetaldehyde inhi-
bition of ethylene biosynthesis in mango fruit. Postharvest Biology and
Technology 8, 153–161.
Burdon, J.N., and Sexton, R. 1990. Fruit abscission and ethylene production
of red raspberry cultivars. Scientia Horticulturae 43, 95–102.
Burg, S.P., and Burg, E.A. 1962. Role of ethylene in fruit ripening. Plant
Physiology 37, 179–189.
Cano, M.P., Ancos, B., Matallana, M.C., Cámara, M., Reglero, G., and
Tabera, J. 1997. Differences among Spanish and Latin American banana
cultivars: Morphological, chemical and sensory characteristics. Food
Chemistry 59, 411–419.
Carrilo-Lopez, A., Cruz-Hernandez, A., Carabez-Trejo, A., Guevara-Lara, F.,
and Paredes-Lopez, O. 2002. Hydrolytic activity and ultrastructural
changes in fruit skins from two prickly pear (Opuntia species) variet-
ies during storage. Journal of Agricultural and Food Chemistry 50,
1681–1685.
Carrilo-Lopez, A., Cruz-Hernandez, A., Guevara-Lara, F., and Paredes-Lopez,
O. 2003. Physico-chemical changes during ripening in storage of two
varieties of prickly pear stored at 18°C. Journal of Food Science and
Technology 40, 461–464.
Carrington, C.M.S. 2011. A frim focus on tropical fruit ripening. Acta
Horticulturae 894, 17–32.
Carrington, C.M.S., and King, R.A.G. 2002. Fruit development and ripening
in Barbados cherry (Malpighia emarginata DC.). Scientia Horticulturae
92, 1–7.
Causse, M., Saliba-Colombani, V., Lesschaeve, I., Duffe, P., Rousselle, P., and
Buret, M. 2001. Genetic analysis of organoleptic quality in fresh mar-
ket tomato. 2. Mapping QTLs for sensory attributes. Theoretical and
Applied Genetics 102, 273–283.
Chen, F.X., Liu, X.H., and Chen, L.S. 2009. Developmental changes in
pulp organic acid concentration and activities of acid-metabolising
enzymes during the fruit development of two loquat (Eriobotrya
japonica Lindl.) cultivars differing in fruit acidity. Food Chemistry 114,
657–664.
Chervin, C., El-Kereamy, A., Roustan, J.P., Latche, A., Lamon, J., and
Bouzayen, M. 2004. Ethylene seems required for the berry develop-
ment and ripening in grape, a non-climacteric fruit. Plant Science 167,
1301–1305.
Chervin, C., Tira-umphon, A., Terrier, N., Zouine, M., Severac, D., and
Roustan, J.P. 2008. Simulation of the grape berry expansion by ethyl-
ene and effects on related gene transcripts over the ripening phase.
Physiologia Plantarum 134, 534–546.

36
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Chyau, C.C., Ko, P.T., Chan, C.H., and Mau, J.L. 2003. Free and glycosidi-
cally bound aroma compounds in lychee (Litchi chinesis Sonn.). Food
Chemistry 80, 387–392.
Clendennen, S.K., and May, G.D. 1997. Differential gene expression in ripen-
ing banana fruit. Plant Physiology 115, 463–469.
Cruz-Hernandez, A., and Paredes-Lopez, O. 2012. Fruit quality: New insights
for biotechnology. Critical Reviews in Food Science and Nutrition 52,
272–289.
Cua, A.U., and Lizada, M.C.C. 1990. Ethylene production in the ‘Carabao’
mango (Mangifera indica L.) fruit during maturation and ripening. Acta
Horticulturae 269, 169–179.
De Souza, M., Silva-Ferreira, K., Paes-Chaves, J.B., and Lopes-Texeira, S.
2008. L-Ascorbic acid, β-carotein and lycopene in papaya fruits (Carica
papaya) with or without physiological skin frackles. Scientia Agricola
65, 246–250.
de Vasconcelos Facundo, H.V., dos Santos Garruti, D., dos Santos Dias, C.T.,
Cordenunsi, B.R., and Lajolo, F.M. 2012. Influence of different banana
cultivars on volatile compounds during ripening in cold storage. Food
Research International 49, 626–633.
Ding, C.K., Chachin, K., Hamauzu, Y., Ueda, Y., and Imahori, Y. 1998. Effects
of storage temperatures on physiology and quality of loquat fruit.
Postharvest Biology and Technology 14, 309–315.
Ding, P., and Tee, Y.K. 2011. Physicochemical characteristics of dabai
(Canarium odontophyllum Miq.) fruit. Fruits 66, 1–6.
Dixon, J., and Hewett, E.W. 2000. Factors affecting apple aroma/flavour
volatile concentration: A review. New Zealand Journal of Crop and
Horticultural Science 28, 155–173.
Downs, C.G., Brady, C.J., Campbell, J.M., and McGlasson, W.B. 1991. Normal
ripening cultivars of Pyrus serotina are either climacteric or non-climac-
teric. Scientia Horticulturae 48, 213–221.
Drury, R., Hortensteiner, S., Donnison, I., Bird, C.R., and Seymour, G.B.
1999. Chlorophyll catabolism and gene expression in the peel of ripen-
ing banana fruits. Physiologia Plantarum 107, 32–38.
Dumville, J.C., and Fry, S.C. 2003. Solubilisation of tomato fruit pectins by
ascorbate: A possible non-enzymatic mechanism of fruit softening.
Planta 217, 951–961.
Durate, O. 2011. Achachairu (Garcinia humilis (Vahl) C.D. Adam). In
Yahia, E.M. (ed.), Postharvest Biology and Technology of Tropical and
Subtropical Fruits. Woodhead Publishing, Cambridge, UK, pp. 48–53.
Egea, I., Barsan, C., Bian, W., Purgatto, E., Latche, A., Chervin, C., Bouzayen,
M., and Pech, J.C. 2010. Chromoplast differentiation: Current status
and perspectives. Plant and Cell Physiology 51, 1601–1611.
Elkashif, M.E., Huber, D.J., and Brecht, J.K. 1989. Respiration and ethylene
production in harvested watermelon fruit: Evidence for non-climacteric
respiratory behaviour. Journal of the American Society for Horticultural
Science 114, 81–85.

37
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Espley, R.V., Brendolise, C., Chagne, D., Kutty-Amma, S., Green, S., Volz, R.,
Putterill, J., Schouten, H.J., Gardiner, S.E., Hellens, R.P., and Allen, A.C.
2009. Multiple repeats of promoter segments causes transcription factor
autoregulation in red apples. Plant Cell 21, 168–183.
Etienne, A., Génard, M., Lobit, P., Mbeguié-A-Mbéguié, D., and Bugaud, C.
2013. What controls fleshy fruit acidity? A review of malate and citrate
accumulation in fruit cells. Journal of Experimental Botany, doi:10.1093/
jxb/ert035.
Etienne, C., Moing, A., Dirlewanger, E., Raymond, P., Monet, R., and Rothan,
C. 2002. Isolation and characterization of six peach cDNAs encoding
key proteins in organic acid metabolism and solute accumulation:
Involvement in regulating peach fruit acidity. Physiologia Plantarum
114, 259–270.
Fait, A., Hanhineva, K., Beleggia, R., Dai, N., Rogachev, I., Nikiforova, V.J.,
Fernie, A.R., and Aharoni, A. 2008. Reconfiguration of the achene and
receptacle metabolic networks during strawberry fruit development.
Plant Physiology 148, 730–750.
Fanciullino, A.L., Cercos, M., Dhuikue-Mayer, C., Froelicher, Y., Talon, M.,
Ollitrault, P., and Morillon, R. 2008. Changes in carotenoids content
and biosynthetic gene expression in juice sacs of four orange varieties
(Citrus sinensis) differing in flesh fruit color. Journal of Agricultural and
Food Chemistry 56, 3628–3638.
Fidler, J.C., and North, C.J. 1967. The effect of storage on the respiration of
apples. I. The effect of temperature and concentration of CO2 and O2 on
the production of CO2 and uptake of O2. Journal of Horticulture Science
42, 189–206.
Fraser, P.D., Kiano, J.W., Truesdale, M.R., Schuch, W., and Bramley, P.M.
1999. Phytoene synthase-2 enzyme activity in tomato does not contrib-
ute to carotenoid synthesis in ripening fruit. Plant Molecular Biology 40,
687–698.
Garcia, E., and Lajola, A.M. 1988. Starch transformation during banana ripen-
ing: The amylase and glucosidase behaviour. Journal of Food Science 53,
1181–1186.
Gardea, A.A., Martínez-Tellez, M.A., Sanchez, A., Baez, M., and Siller,
J.H. 1994. Postharvest weight loss of Flame Seedless clusters. In
International Symposium on Table Grape Production, Anaheim, CA,
pp. 203–206.
Gariglio, N., Juan, M., Castillo, A., Almela, V., and Agustí, M. 2002.
Morphological, histological and physiological study of purple spot of
loquat fruit. Scientia Horticulturae 92, 255–263.
Gariglio, N.F., Reig, C., and Agusti, M. 2008. Assimilate partitioning between
the flesh and the rind is responsible for purple spot in loquat fruit.
Journal of Horticultural Science and Biotechnology 83, 37–42.
Giovannoni, J. 2001. Molecular biology of fruit maturation and ripening.
Annual Review of Plant Physiology and Plant Molecular Biology 52,
725–749.

38
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Gonzalez, L., Lafuente, M.T., and Zacarias, L. 2004. Maturation of loquat fruit
(Eriobotrya japonica Lindl.) under Spanish growing conditions and its
postharvest performance. Options Mediterran 58, 171–179.
Goulao, L.F., and Oliveira, C.M. 2008. Cell wall modifications during fruit rip-
ening: When a fruit is not fruit. Trends in Food Science and Technology
19, 4–25.
Graham, O.S., Mohammed, M., and Wickham, L.D. 2004. Effects of heat
treatments on the quality of miniature golden apples (Spondias cytherea
Sonn) during low temperature storage. International Journal of Food,
Agriculture and Environment 2, 43–48.
Gurrieri, F., Audergon, J.M., Albagnac, G., and Reich, M. 2001. Soluble sug-
ars and carboxylic acids in ripe apricot fruit as parameters for distin-
guishing different cultivars. Euphytica 117, 183–189.
Hamauzu, Y., Chachin, K., Ding, C.K., and Kurooka, H. 1999. Differences in
surface color, flesh firmness, physiological activity, and some compo-
nents of loquat fruit picked at various stages of maturity. Journal of the
Japanese Society for Horticultural Science 65, 859–865.
Hardy, P.J. 1968. Metabolism of sugars and organic acids in immature grape
berries. Plant Physiology 43, 224–228.
Hasegawa, P.N., Faria, A.F., Mercadante, A.Z., Chagas, E.A., Pio, R., Lajolo,
F.M., Cordenunsi, B.R., and Purgatto, E. 2010. Chemical composition of
five loquat cultivars planted in Brazil. Ciencia e Tecnologia de Alimentos
30, 552–559.
Hernandez, M.S., Barrera, J.A., Martinez, O., and Fernandez-Trujillo, J.P.
2009. Postharvest quality of araza fruit during low temperature storage.
LWT Food Science and Technology 42, 879–884.
Hernandez, M.S., Martinez, O., and Fernandez-Trujillo, J.P. 2007. Behavior
of araza fruit quality traits during growth, development and ripening.
Scientia Horticulturae 111, 220–227.
Hirai, M. 1980. Sugar accumulation and development of loquat fruit. Journal
of the Japanese Society of Horticultural Science 49, 347–353.
Huber, D.J. 1984. Strawberry fruit softening: The potential roles of polyur-
onoids and hemicelluloses. Journal of Food Science 49, 1310–1315.
Hulme, A.C. 1958. Some aspects of the biochemistry of apple and pear fruit.
Advances in Food Research 8, 297–413.
Hulme, A.C. 1970. The Biochemistry of Fruits and Their Products, vol. I.
Academic Press, London, p. 620.
Ilahy, R., Hdider, C., Lenucci, M.S., Tlili, I., and Dalessandro, G. 2011.
Antioxidant activity and bioactive compound changes during fruit ripen-
ing of high-lycopene tomato cultivars. Journal of Food Composition and
Analysis 24, 588–595.
Ishimaru, M., and Kobayashi, S. 2002. Expression of a xyloglucan endo-
transglycosilase gene is closely related to grape berry softening. Plant
Science 162, 621–628.
Ismael, A.A., and Render, W.T. 1969. Evidence of a respiratory climacteric in
highbush and lowbush blueberry fruit. HortScience 4, 342–344.

39
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Itai, A., Tanabe, K., Tamura, F., and Tanaka, T. 2000. Isolation of cDNA clones
corresponding to genes expressed during fruit ripening in Japanese
pear (Pyrus pyrifolia Nakai): involvement of the ethylene signal trans-
duction pathway in their expression. Journal of Experimental Botany 51,
1163–1166.
Iwaoka, W., Hagi, Y., Umano, K., and Shibamoto, T. 1994. Volatile chemicals
identified in fresh and cooked breadfruit. Journal of Agricultural and
Food Chemistry 42, 975–976.
Jiang, J.P., Su, M.X., and Lee, P.M. 1986. The production and physiological
effects of ethylene during ontogeny and after harvest of litchi fruits.
Acta Phytophysiologia Sinica 12, 95–103.
Jiang, W.B., and Sheng, Q. 2003. Effects of 1-methylcyclopropene and giberel-
lic acid on ripening of jujube (Zizyphus jujuba M) in relation to quality.
Journal of the Science of Food and Agriculture 84, 31–35.
Jordan, M.J., Tandon, K., Shaw, P.E., and Goodner, K.L. 2001. Aromatic profile
of aqueous banana essence and banana fruit by gas ­chromatography–
mass spectrometry (GC–MS) and gas chromatography–olfactometry
(GC–O). Journal of Agricultural and Food Chemistry 49, 4813–4817.
Joyce, D.C., and Li, J.R. 2003. Postharvest characteristics of red bayberry.
In Proceedings of the Australasian Postharvest Horticulture Conference,
Brisbane, Australia, October 1–3, pp. 125–226.
Kader, A. 2002. Loquat: recommendations for maintaining postharvest qual-
ity. http://postharvest.ucdavis.edu/produce/fruit/loquat.html.
Kader, A.A., and Arpaia, M.L. 2001. Avocado: recommendations for main-
taining postharvest quality. http://postharvest.ucdavis.edu/produce/­
producefacts/fruit/ avocado.html.
Kader, A.A., Li, Y., and Chordas, A. 1982. Postharvest respiration, ethyl-
ene production, and compositional changes in Chinese jujube fruits.
HortScience 17, 678–679.
Kalra, S.K., and Tandon, D.K. 1983. Ripening behavior of Dashehari mango in
relation to harvest period. Scientia Horticulturae 19, 263–269.
Katz, E., Lagunes, P.M., Riov, J., Weiss, D., and Goldschmidt, E.E. 2004.
Molecular and physiological evidence suggests the existence of a sys-
tem II-like pathway of ethylene production in non-climacteric citrus
fruit. Planta 219, 243–252.
Kays, S. 1991. Postharvest Physiology of Perishable Plant Products. Van
Nostrand Reinhold, New York.
Kays, S.J., and Hayes, M.J. 1978. Induction of ripening in the fruits of bitter
gourd by ethylene. Tropical Agriculture 55, 167–171.
Ketsa, S., and Daengkanit, T. 1998. Physiological changes during posthar-
vest ripening of durian fruit (Durio zibethinus Murray). Journal of
Horticultural Science and Biotechnology 73, 575–577.
Klee, H.J. 2002. Control of ethylene-mediated processes in tomato at the level
of receptors. Journal of Experimental Botany 53, 2057–2063.

40
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Kliewer, W.M. 1971. The effect of day temperature and light intensity on con-
centration of malic and tartaric acids in Vitis vinifera L. grapes. Journal
of the American Society for Horticultural Science 96, 372–377.
Kobayashi, S., Goto-Yamamoto, N., and Hirochika, H. 2004. Retrotransposon-
induced mutations in grape skin color. Science 304, 982.
Krishnamurthy, S., and Subramanyam, H. 1970. Respiratory climacteric and
chemical changes in the mango fruit Mangifera indica L. Journal of the
American Society for Horticultural Science 95, 333–337.
Kurz, C., Carle, R., and Schieber, A. 2008. HPLC-DAD-MS characterization
of carotenoids from apricots and pumpkins for the evaluation of fruit
product authenticity. Food Chemistry 110, 522–530.
Lakshminarayana, S., and Estrella, I.B. 1978. Postharvest respiratory behav-
iour of tuna (prickly pear) fruit (Opuntia robusta Mill.). Journal of
Horticultural Science 53, 327–330.
Lam, P.F., and Wan, C.K. 1983. Climacteric nature of the carambola (Averrhoa
carambola L.) fruit. Pertanika 6, 44–47.
Lannetta, P.P.M., Laarhoven, L.I., Medina-Escobar, N., James, E.K., McManus,
M.T., Davies, H.V., and Harren, F.J.M. 2006. Ethylene and carbon diox-
ide production by developing strawberries show a correlative pattern
that is indicative of ripening climacteric fruit. Physiologia Plantarum
127, 247–259.
Lazan, H., Ali, Z.M., Soh, J.S., and Talkah, Z. 1993. The biochemical basis of
differential ripening in mango. Acta Horticulturae 341, 500–509.
Lelievre, J.M., Latche, A., Jones, B., Bouzayen, M., and Pech, J.C. 1997.
Ethylene and fruit ripening. Physiologia Plantarum 101, 727–739.
Li, X., Xu, C., Korban, S.S., and Chen, K. 2010. Regulatory mechanisms of
textural changes in ripening fruits. Critical Reviews in Plant Sciences
29, 222–243.
Lipe, J.A. 1978. Ethylene in fruits of blackberry and rabbiteye blueberry.
Journal of the American Society for Horticultural Science 103, 76–77.
Lo Bianco, R., and Rieger, M. 2002. Partitioning of sorbitol and sucrose catab-
olism within peach fruit. Journal of the American Society for Horticulture
Science 127, 115–121.
Lu, X.-P., Liu, Y.-Z., Zhou, G.-F., Wei, Q.-J., Hu, H.-J., and Peng, S.-A. 2011.
Identification of organic acid-related genes and their expression profiles
in two pear (Pyrus pyrofolia) cultivars with difference in predominant
acid type at fruit ripening stage. Scientia Horticulturae 129, 680–687.
Lyons, J.M., McGlasson, W.B., and Pratt, H.K. 1962. Ethylene production,
respiration and internal gas concentrations in cantaloupe fruits at vari-
ous stages of maturity. Plant Physiology 37, 31–36.
Malik, S.K., Kumar, S., and Bansal, K.C. 2014. Genetic diversity of tropical
fruit. In Nath, P., Bouzayen, M., Mattoo, A.K., and Pech, J.C. (eds.),
Fruit Ripening: Physiology, Signaling and Genomics. CABI, Oxfordshire,
UK, pp. 217–227.

41
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Manenoi, A., Bayogan, E.R.V., Thumdee, S., and Paull, R.E. 2007. Utility of
1-methylcyclopropene as a papaya postharvest treatment. Postharvest
Biology and Technology 44, 55–62.
Marei, N., and Crane, J.C. 1971. Growth and respiratory response of fig
(Ficus  carica L. cv. Mission) fruits to ethylene. Plant Physiology 48,
249–254.
Marin-Rodriguez, M.C., Orchard, J., and Seymour, G.B. 2002. Pectate lyases,
cell wall degradation and fruit softening. Journal of Experimental Botany
53, 2115–2119.
Mattoo, A.K., and Modi, V.V. 1969. Biochemical aspects of ripening and chill-
ing injury in mango fruit. In Proceedings of International Conference on
Tropical and Subtropical Fruit, London, pp. 111–115.
Maxie, E.C., Catlin, P.B., and Hartman, H.T. 1960. Respiration and ripening of
olive fruits. Proceedings of the American Society for Horticultural Science
75, 275–291.
McCarthy, A.I., Palmer, J.K., Shaw, C.P., and Anderson, E.E. 1963. Correlation
of gas chromatographic data for flavour profiles of fresh banana fruits.
Journal of Food Science 28, 379–384.
Medina-Suarez, R., Manning, K., Fletcher, J., Aked, J., Bird, C.R., and
Seymour, G.B. 1997. Gene expression in the pulp of ripening bananas
(two-dimensional sodium dodecyl sulfate-polyacrylamide gel electro-
phoresis of in vitro translation products and cDNA cloning of 25 differ-
ent ripening-related mRNAs). Plant Physiology 115, 453–461.
Mendoza, D.B., Pantastico, E., and Javier, F.B. 1972. Storage and handling
of rambutan (Nephelium lappaceum L.). Philippines Agriculture 55,
322–332.
Moore, S., Vrebalov, J., Payton, P., and Giovannoni, J. 2002. Use of genom-
ics tools to isolate key ripening genes and analyse fruit maturation in
tomato. Journal of Experimental Botany 53, 2023–2030.
Morandi, B., Grappadelli, L.G., Rieger, M., and Lo Bianco, R. 2008.
Carbohydrate availability affects growth and metabolism in peach fruit.
Physiologia Plantarum 133, 229–241.
Mota, W.F., Salomao, L.C.C., Perira, M.C.T., and Cecon, P.R. 2002. The influ-
ence of the postharvest treatment with calcium in jaboticaba fruits con-
servation. Revista Brasieira de Fruticultura 24, 49–52.
Nam, Y.W., Tichit, L., Leperlier, M., Cuerq, B., Marty, I., and Lelievre, J.M.
1999. Isolation and characterization of mRNA differentially expressed
during ripening of wild strawberry (Fragaria vesca L.) fruits. Plant
Molecular Biology 39, 629–636.
Nerd, A., Gutman, F., and Mizrahi, Y. 1999. Ripening and postharvest behav-
iour of fruits of two Hylocereus species (Cactaceae). Postharvest Biology
and Technology 17, 39–45.
Nerd, A., and Mizrahi, Y. 1999. The effect of ripening stage on fruit quality
after storage of yellow pitaya. Postharvest Biology and Technology 15,
99–105.

42
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Nogueira, J.M., Fernandes, P.J., and Nascimento, A.M. 2003. Composition

of volatiles of banana cultivars from Madeira Island. Phytochemical
Analysis 14, 87–90.
Nunan, K.J., Davies, C., Robinson, S.P., and Fincher, G.B. 2001. Expression
patterns of cell wall modifying enzymes during grape berry develop-
ment. Planta 214, 257–264.
Oslund, C., and Davenport, T. 1983. Ethylene and carbon dioxide in ripening
fruit of Averrhoa carambola. HortScience 18, 229–230.
Palma, J.M., Corpas, F.J., and del Rio, L.A. 2011. Proteomics as an approach
to the understanding of the molecular physiology of fruit development
and ripening. Journal of Proteomics 74, 1230–1243.
Pareek, S., and Kitinoja, L. 2011. Aonla (Emblica officinalis Gaertn.). In
Yahia, E.M. (ed.), Postharvest Biology and Technology of Tropical
and Subtropical  Fruits, Woodhead Publishing Ltd., Cambridge, UK,
pp. 65–97.
Pareek, S., and Yahia, E.M. 2013. Postharvest biology and technology of ber
fruit. Horticultural Reviews 41, 201–240.
Pareek, S., Benkeblia, N., Janick, J., Cao, S., and Yahia, E.M. 2014. Postharvest
physiology and technology of loquat (Eriobotria japonica Lindl.) fruit.
Journal of the Science of Food and Agriculture 94, 1495–1504.
Pareek, S., Kitinoja, L., Kaushik, R.A., and Paliwal, R. 2009. Postharvest physi-
ology and storage of ber. Stewart Postharvest Review 5, 1–10.
Paul, V., Pandey, R., and Srivastava, G.C. 2012. The fading distinctions
between classical patterns of ripening in climacteric and non-climacteric
fruit and the ubiquity of ethylene: an overview. Journal of Food Science
and Technology 49, 1–21.
Paull, R.E., and Ketsa, S. 2004. Mangosteen. In Gross, K.C., Wang, C.Y.,
and Saltveit, M.E. (eds.), The Commercial Storage of Fruits, Vegetables
and Florists and Nursery Stocks. USDA Handbook No. 66 revised. U.S.
Department of Agriculture, Agricultural Research Service, Washington,
DC. http://www.ba.ars.usda./hb66/092mangosteen.pdf (accessed
February 23, 2015).
Payasi, A., and Sanwal, G.G. 2005. Biochemistry of fruit ripening. Indian
Journal of Agricultural Biochemistry 18, 51–60.
Payasi, A., and Sanwal, G.G. 2009. Molecular mechanism of ethylene signal
transduction in fruits. Indian Journal of Agricultural Biochemistry 22,
1–11.
Pech, J.C., Bouzayen, M., and Latche, A. 2008. Climacteric fruit ripening:
ethylene-dependent and independent regulation of ripening pathways
in melon fruit. Plant Science 175, 114–120.
Pech, J.C., Purgatto, E., Bouzayen, M., and Latche, A. 2012. Ethylene and
fruit ripening. Annual Plant Reviews 44, 275–304.
Pech, J.C., Purgatto, E., Girardi, C.L., Rombaldi, C.V., and Latche, A. 2013.
Current challenges in postharvest biology of fruit ripening. Current
Agricultural Science and Technology 19, 1–18.

43
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Pérez, A.G., Cert, A., Rios, J.J., and Olias, J.M. 1997. Free and glycosidi-
cally bound volatile compounds from two banana cultivars: Valery
and Pequena Enana. Journal of Agricultural and Food Chemistry 45,
4393–4397.
Perkins-Veazie, P., and Nonneeke, G. 1992. Physiological changes during rip-
ening of raspberry fruit. HortScience 27, 331–333.
Perkins-Veazie, P., Collins, J.K., Pair, S.D., and Roberts, W. 2001. Lycopene
content differs among red-fleshed watermelon cultivars. Journal of the
Science of Food and Agriculture 81, 983–987.
Pontes, M., Pereira, J., and Câmara, J.S. 2012. Dynamic headspace solid-
phase microextraction combined with one-dimensional gas chroma-
tography-mass spectrometry as a powerful tool to differentiate banana
cultivars based on their volatile metabolite profile. Food Chemistry 134,
2509–2520.
Pratt, H.K., and Goeschl, J.D. 1968. The role of ethylene in fruit ripening. In
Wightman, F., and Setterfield, G. (eds.), Biochemistry and Physiology of
Plant Growth Substances. Runge, Ottawa, pp. 1295–1302.
Pratt, H.K., and Mendoza, D.B. 1980. Fruit development and ripening of the
star apple (Chrysophyllum cainito L.). HortScience 15, 721–722.
Pratt, H.K., and Reid, M.S. 1974. Chinese gooseberry: Seasonal patterns in
fruit growth and maturation, ripening, respiration and role of ethylene.
Journal of the Science of Food and Agriculture 25, 747–753.
Pratt, H.K., and Reid, M.S. 1976. The tamarillo: Fruit growth and maturation,
ripening, respiration, and the role of ethylene. Journal of the Science of
Food and Agriculture 27, 399–404.
Prinsi, B., Negri, A.S., Fedeli, C., Morgutti, S., Negrini, N., Cocucci, M., and
Espen, L. 2011. Peach fruit ripening: A proteomic comparative analysis
of the mesocarp of two cultivars with different flesh firmness at two
ripening stages. Phytochemistry 72, 1251–1262.
Reid, M.S. 1975. The role of ethylene in the ripening of some unusual fruits.
In Facteurs et regulation de la maturation des fruits, no. 238. CNRS,
Paris, pp. 177–182.
Reig, C., Martinez-Fuentes, A., Juan, M., Gariglio, N., Marti, G., Mesejo, C.,
and Agusti, M. 2007. Tecnicasparaanticipar la recoleccion del fruto del
nisperojapones (Eriobotrya japonica Lindl.). In XI Congress National
SECH, Spain, abstract 4D01.
Rogiers, S.Y., Mohan-Kumar, G.N., and Knowles, N.R. 1998. Regulation of
ethylene production and ripening by Saskatoon (Amelanchier alnifolia
Nutt.) fruit. Canadian Journal of Botany 76, 1743–1754.
Roy, S.K. 1975. Studies of bael fruit (Aegle marmelos Carrea)—changes in
constituents during development, ripening and storage and process-
ing of the fruit. PhD thesis, Bidhan Chandra Krishi Vishva Vidyalaya,
Kalyani, West Bengal, India.
Rygg, G.L. 1975. Date Development, Handling and Packing in the United
States. Agricultural Handbook No. 482. U.S. Department of Agriculture,
Agricultural Research Service, Washington, DC, p. 56.

44
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Salmon, B., Martin, G.J., Remaud, G., and Fourel, F. 1996. Compositional and
isotopic studies of fruit flavours. Part I. The banana aroma. Flavour and
Fragrance Journal 11, 353–359.
Saltveit, M.E. 1977. Carbon dioxide, ethylene, and color development in rip-
ening mature green bell peppers. Journal of the American Society for
Horticultural Science 102, 523–525.
Sampaio, S.A., Bora, P.S., Holschuh, H.J., and Silva, S.M. 2007. Postharvest
respiratory activity and changes in some chemical constituents dur-
ing maturation of yellow mombin (Spondias mombin) fruit. Ciencia e
Technologia de Alimentos 27, 1–9.
Santos, A.F. dos, Silva, S.M., and Alves, R.E. 2006. Armezenamento de pitanga
sob atmosfera modificada refrigeracao: I-transformacoes quimicas em
pos-colheita. Revista Brasieira de Fruticultura 28, 36–41.
Saradhuldhat, P., and Paull, R.E. 2007. Pineapple organic acid metabolism
and accumulation during fruit development. Scientia Horticulturae 112,
297–303.
Selvaraj, Y., and Pal, D.K. 1989. Biochemical changes during the ripening
of jackfruit (Artocarpus heterophyllus L.). Journal of Food Science and
Technology 25, 304–307.
Seymour, G.B., Manning, K., Eriksson, E.M., Popovich, A.H., and King, G.J.
2002. Genetic identification and genomic organization of factors affect-
ing fruit texture. Journal of Experimental Botany 5, 2065–2071.
Shen, L., Sheng, J.P., Niu, J.S., and Liu, Q.X. 2004. Changes of respiration
and ethylene production and effects of 1-MCP during the fermentation
softening of Chinese winter jujube fruit. Journal of China Agricultural
University 9, 36–39.
Shiota, H. 1993. New esteric compounds in the volatiles of banana fruit (Musa
sapientum L.). Journal of Agricultural and Food Chemistry 41, 2056–2062.
Sinesio, F., Cammareri, M., Moneta, E., Navez, B., Peparaio, M., Causse,
M., and Grandillo, S. 2010. Sensory quality of fresh French and Dutch
market tomatoes: A preference mapping study with Italian consumers.
Journal of Food Science 75, 55–67.
Singh, S.P., and Pal, R.K. 2008. Response of climacteric-type guava (Psidium
guajava L.) to postharvest treatment with 1-MCP. Postharvest Biology
and Technology 47, 307–314.
Siriphanich, J., Abdulla, H., Kosittrakun, M., and Lizada, M.C.C. 1994.
Physiology. In Nanthachai, S. (ed.), Durian Fruit Development,
Postharvest Physiology, Handling and Marketing in ASEAN. ASEAN
Food Handling Bureau, Kuala Lumpur, pp. 48–57.
Sivakumar, D., Naudé, Y., Rohwer, E., and Korsten, L. 2008. Volatile com-
pounds, quality attributes, mineral composition and pericarp structure
of South African litchi export cvs. Mauritius and McLean’s Red. Journal
of the Science of Food and Agriculture 88, 1074–1081.
Sweetman, C., Deluc, L.G., Cramer, G.R., Ford, C.M., and Soole, K.L. 2009.
Regulation of malate metabolism in grape berry and other developing
fruits. Phytochemistry 70, 1329–1344.

45
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Thompson, A.J., Tor, M., Barry, C.S., Vrebalov, J., Orfila, C., Jarvis, M.C.,
Giovannoni, J.J., Grierson, D., and Seymour, G.B. 1999. Molecular and
genetic characterization of a novel pleiotropic tomato-ripening mutant.
Plant Physiology 120, 383–389.
Tian, M., Sheng, C.C., and Li, Y. 1987. Changes of ethylene synthesis, activity
of polyphenol oxidase, permeability of plasma membrane of Dutch pear
at low temperature. Acta Botanic Sinica 29, 614–619.
Toivonen, P.M.A., and Brummell, D.A. 2008. Biochemical bases of appear-
ance and texture changes in fresh-cut fruit and vegetables. Postharvest
Biology and Technology 48, 1–14.
Tongdee, S.C., Suwanakul, A., and Neamprem, S. 1990. Durian fruit ripen-
ing and the effect of variety, maturity stage at harvest, and atmospheric
gases. Acta Horticulturae 269, 323–334.
Topuz, A., and Ozdemir, F. 2007. Assessment of carotenoids, capsaicinoids
and ascorbic acid composition of some selected pepper cultivars
(Capsicum annuum L.) grown in Turkey. Journal of Food Composition
and Analysis 20, 596–602.
Tressl, R., and Jennings, W.G. 1972. Production of volatile compounds in
the ripening banana. Journal of Agricultural and Food Chemistry 20,
189–192.
Trinchero, G.D., Sozzi, G.O., Cerri, A.M., Vilella, F., and Fraschina, A.A. 1999.
Ripening-related changes in ethylene production, respiration rate and
cell-wall enzyme activity in goldenberry (Physalis peruviana L.), a sola-
naceous species. Postharvest Biology and Technology 16, 139–145.
Tucker, G.A. 1993. Introduction. In Seymour, G.B., Taylor, J.E., and Tucker,
G.A., Biochemistry of Fruit Ripening. Springer, London, UK. pp. 1–52.
Tucker, G.A., and Grierson, D. 1987. Fruit ripening. In Davies, D. (ed.), The
Biochemistry of Plants, vol. 12. Academic Press, New York, pp. 265–319.
Ulrich, D., Hoberg, E., Rapp, A., and Kecke, S. 1997. Analysis of strawberry
flavor—discrimination of aroma types by quantification of volatile com-
pounds. Zeitschrift fur Lebensmittel-Untersuchung und Forschung A 205,
218–223.
Velasquez de Klimo, I. 2006. Manejo pos cosecha del nance (Byrsonima crassi-
folia (L.) HBK). Ministerio de Agricultura y Ganaderia, IICA Frutales,
Programa Nacional de Frutales, San Salvador, El Salvador.
Vermeir, S., Hertog, M.L.A.T.M., Vankerschaver, K., Swennen, R., Nicolai,
B.M., and Lammertyn, J. 2009. Instrumental based flavour characteriza-
tion of banana fruit. LWT Food Science and Technology 42, 1647–1653.
Vicente, A.R., Ortugno, C., Powell, A.L., Greve, L.C., and Labavitch, J.M.
2007a. Temporal sequence of cell wall disassembly events in developing
fruits. 1. Analysis of raspberry (Rubus idaeus). Journal of Agricultural
and Food Chemistry 55, 4119–4124.
Vicente, A.R., Ortugo, C., Rosli, H., Powell, A.L., Greve, L.C., and Labavitch,
J.M. 2007b. Temporal sequence of cell wall disassembly events in devel-
oping fruits. 2. Analysis of blueberry (Vaccinium species). Journal of
Agricultural and Food Chemistry 55, 4125–4130.

46
 Ripen in g P hy s i o l o g y : A n Ov e r v i e w

Vicente, A.R., Powell, A., Greve, L.C., and Labavitch, J.M. 2007c. Cell wall disas-
sembly events in boysenberry (Rubus idaeus L. × Rubus ursinus Cham. &
Schldl.) fruit development. Functional Plant Biology 34, 614–623.
Vicente, A.R., Saladie, M., Rose, J.K.C., and Labavitch, J.M. 2007d. The link-
age between cell wall metabolism and fruit softening: looking to the
future. Journal of the Science of Food and Agriculture 87, 1435–1448.
Vrebalov, J., Ruezinsky, D., Padmanabhan, V., White, R., Medrano, D., Drake, R.,
Schuch, W., and Giovannoni, J. 2002. A MADS-box gene necessary for fruit
ripening at the tomato ripening-inhibitor (rin) locus. Science 296, 343–346.
Wade, N.L., Kavanagh, E.E., Hockley, D.G., and Brady, C.J. 1992. Relationship
between softening and the polyuronides in ripening banana fruits.
Journal of the Science of Food and Agriculture 60, 61–68.
Walsh, C.S., Popenoe, J., and Solomos, T. 1983. Thornless blackberry is a
­climacteric fruit. HortScience 18, 482–483.
Walker, A.R., Lee, E., Bogs, J., McDavid, D.A.J., Thomas, M.R., and Robinson,
S.P. 2007. White grapes arose through the mutation of two similar and
adjacent regulatory genes. Plant Journal 49, 772–785.
Wang, T., Gonzalez, A.R., Gbur, E.E., and Aselage, J.M. 1993. Organic acid
changes during ripening of processing peaches. Journal of Food Science
58, 631–632.
Warren, O. 2009. Quality of carambola fruit (Averrhoa carambola L.) as
affected by harvest maturity, postharvest wax coating, ethylene, and
1-methylcyclopropene. MSc thesis, Horticultural Sciences Department,
University of Florida, Gainesville.
Waters, D.L.E., Holton, T.A., Albett, E.M., Lee, L.S., and Henry, R.J. 2005.
cDNAs microarrays analysis of developing grape (Vitis vinifera cv.
Shiraz) berry skin. Functional and Integrative Genomics 5, 40–58.
Watkins, C.B. 2002. Ethylene biosynthesis: Mode of action, consequences,
and control. In Knee, M. (ed.), Fruit Quality and Its Biological Basis,
Sheffield Academic Press, Sheffield, pp. 180–224.
Watson, R., Wright, C.J., McBurney, T., Taylor, A.J., and Linforth, R.S.T. 2002.
Influence of harvest date and light integral on the development of straw-
berry flavour compounds. Journal of Experimental Botany 53, 2121–2129.
Weaver, R.J., and Singh, I.S. 1978. Occurrence of endogenous ethylene and
effect of plant regulators on ethylene production in the grapevine.
American Journal of Enology and Viticulture 29, 282–285.
White, P.J. 2002. Recent advances in fruit development and ripening: An over-
view. Journal of Experimental Botany 53, 1995–2000.
Whiting, G.C. 1970. Sugars. In Hulme, A.C. (ed.), The Biochemistry of Fruits
and Their Products, vol. I. Academic Press, London, pp. 1–620.
Wu, Y., Pan, Q., Qu, W., and Duan, C. 2009. Comparison of volatile profiles
of nine litchi (Litchi chinensis Sonn.) cultivars from southern China.
Journal of Agricultural and Food Chemistry 57, 9676–9681.
Yahia, E.M. 2004. Sapodilla and related fruits. In Gross, K.C., Wang, C.Y.,
and Saltveit, M.E. (eds.), The Commercial Storage of Fruits, Vegetables
and Florists and Nursery Stocks. USDA Handbook No. 66 revised.

47
P OST H AR V EST RI P ENING P H Y SIOLOG Y

U.S.  Department of Agriculture, Agricultural Research Service,

Washington, DC. http://www.ba.ars.usda./hb66/092mangosteen.pdf
(accessed February 23, 2015).
Yamaguchi, S., Yoshikawa, T., Ikeda, S., and Ninomiya, T. 1970. Studies on
the taste of some sweet substances. Part I. Measurement of the relative
sweetness. Agricultural and Biological Chemistry 34, 181–186.
Yamaki, S. 1984. Isolation of vacuoles from immature apple fruit flesh and
compartmentation of sugars, organic acids, phenolic compounds and
amino acids. Plant and Cell Physiology 25, 151–166.
Yamaki, Y.T. 1989. Organic acids in the juice of citrus fruits. Journal of the
Japanese Society for Horticultural Science 58, 587–594.
Yang, L.T., Xie, C.Y., Jiang, H.X., and Chen, L.S. 2011. Expression of six
malate-related genes in pulp during the fruit development of two loquat
(Eriobotrya japonica) cultivars differing in fruit acidity. African Journal
of Biotechnology 10, 2414–2422.
Yelle, S., Chetelat, R.T., Dorais, M., Deverna, J.W., and Bennett, A.B. 1991.
Sink metabolism in tomato fruit. IV. Genetic and biochemical analysis
of sucrose accumulation. Plant Physiology 95, 1026–1035.
Zhang, X., Yazaki, J., Sundaresan, A., Cokus, S., Chan, S., Chen, H., Henderson,
I., Shinn, P., Pellegrini, M., Jacobsen, S., and Ecker, J.R. 2006. Genome-
wide high resolution mapping and functional analysis of DNA methyla-
tion in Arabidopsis. Cell 126, 1189–1201.
Zhao, Y.F., Lin, H.T., Lin, J.F., Chen, S.J., and Xi, Y.F. 2005. Changes of respi-
ration rate, cell membrane permeability and quality in postharvest lon-
gan fruit. Journal of Fujian Agriculture and Forest University (Natural
Science Edition) 34, 263–268.
Zhou, C.H., Xu, C.J., Sun, C.D., Li, X., and Chen, K.S. 2007. Carotenoids in
white- and red-fleshed loquat fruits. Journal of Agricultural and Food
Chemistry 55, 7822–7830.

48
Chapter 2

Postharvest
Physiology of Fruits
and Vegetables
Peter M.A. Toivonen
Agriculture and Agri-Food Canada
Summerland, British Columbia, Canada

Abstract 50
2.1 Introduction 51
2.2  Classification of Fruits and Vegetables Based on
Physiological Characteristics 51
2.2.1  Ethylene Biology 51
2.2.2  Respiratory Characteristics: Climacteric and
Nonclimacteric 56
2.2.3  Developmental or Maturity Stage at Harvest 56
2.2.4  Tolerance to Low Temperatures 58
2.3  Factors Affecting Physiology of Fruits and Vegetables in
Postharvest Systems 60
2.3.1 Temperature 60
2.3.2 Humidity 62
2.3.3  Atmospheric Modification 64
2.3.4  Abiotic Stresses 65

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

2.4  Physiological Changes Occurring during Postharvest

Handling or Storage 66
2.4.1  Depletion of Respiratory Substrate 66
2.4.2  Hormonal Effects 68
2.4.3  Membrane Alterations 71
2.5  Conclusions and Future Perspectives 71
References 72

Abstract
This chapter covers a wide range of issues pertaining to the many aspects
of fruit and vegetable tissue physiology and changes that occur during
postharvest handling and storage. Understanding the specific ethyl-
ene biology, respiration behavior, and developmental or maturity stage
at harvest can provide great insight to postharvest quality retention
characteristics of a fruit or vegetable. Ethylene itself is one of the most
important factors determining quality retention, with different fruit and
vegetable classes having differing production rates and sensitivity to the
levels of ethylene present. Numerous disorders and quality defects can
be attributed to exposures to ethylene in postharvest systems, and these
are discussed. Respiration behaviors can be separated into two classes:
(1) climacteric and (2) nonclimacteric. The classification of respira-
tory behavior will have an impact on postharvest approaches required
for optimal handling of a particular fruit or vegetable in question. The
anatomical structure and developmental maturity of a fruit or vegetable
can range from an immature vegetable tissue all the way to a ripe fruit
tissue, and this will also determine which postharvest procedures will
be successful for handling and storage life potential. The importance of
temperature, humidity, and atmosphere management on quality reten-
tion is discussed in a mechanistic manner to explain the physiological
responses to these postharvest interventions. Finally, endogenous and
exogenously applied phytohormones are important to signaling meta-
bolic shifts that influence quality retention, and these implications are
explored. The discussion in this chapter is intended to provide a basic
and mechanistic understanding of how and why quality changes occur
in postharvest handling of fruits and vegetables that have contrasting
physiological characteristics. The discussion culminates in discussion
on improving quality retention of fruits and vegetables through multi-
disciplinary cooperation of plant breeders, physiologists, biochemists,
and molecular biologists, leading to more robust solutions to postharvest
problems.

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

2.1 Introduction
Development of successful harvest, handling, storage, and distribution
protocols for fruits and vegetables has relied on the understanding of the
physiology of each type of produce and how that physiology responds to
the postharvest environment to which it is exposed (Toivonen and Hodges,
2011). Because of limitations in produce handling systems, many different
types of fruits and vegetables are held in common rooms or containers,
and this has led to issues of incompatibility of produce having different
physiologies. This incompatibility is expressed as shorter storage or shelf
life, development of off-flavors, and accumulation of bitter compounds
(Thompson et al., 1996). Another consequence of common storage areas
is that some fruits and vegetables have differing tolerances to low tempera-
tures, and therefore some fruits or vegetables held in common-use, low-
temperature storage might suffer chilling injuries (Thompson et al., 1996;
Cantwell, 2002). This phenomenon continues to exist and can still today
be visualized in retail produce displays as pitting with associated decay in
cantaloupes and summer squash or as sheet pitting in green peppers (per-
sonal observation). The challenge facing postharvest science and technol-
ogy today is the development of innovative strategies to resolve problems
associated with differing fruit and vegetable physiologies in the real-world
harvest, storage, distribution, and retail systems.
This chapter is devoted to providing an overview of the fundamen-
tal aspects of fruit and vegetable postharvest physiologies to allow a better
appreciation for challenges faced in their handling. Understanding of the
fundamental principles will provide grounded principles that will hopefully
be the basis of new, innovative strategies for harvest, storage, handing, and
distribution of these living and very perishable products.

2.2  Classification of Fruits and Vegetables

Based on Physiological Characteristics
There are a number of aspects of fruit or vegetable physiology that can
influence postharvest quality outcomes. These aspects include ethylene
biology, respiration, the developmental or maturity stage at harvest, and
sensitivity to low temperature.

2.2.1  Ethylene Biology

Ethylene is arguably the most important molecule in the postharvest han-
dling of fruits and vegetables, having a wide range of effects that lead to qual-
ity loss (Saltveit, 1999). The reason for ethylene’s prominence on postharvest

51
P OST H AR V EST RI P ENING P H Y SIOLOG Y

­ andling is that it is a plant growth regulator that is naturally produced with

h
ripening of climacteric fruit and generally in response to either abiotic or
biotic stresses (Saltveit, 1999). However, another feature of postharvest han-
dling is that fruits and vegetables are usually held in containers, chambers,
or rooms, and this allows any ethylene produced by the produce to accumu-
late. Quite often, fruits and vegetables that produce ethylene or are sensi-
tive to ethylene are handled alongside each other, leading to quality losses
in sensitive produce in that containment (Saltveit, 1999). While thresholds
to ethylene sensitivity have been generally thought to be around 0.1 µl L –1,
there is evidence suggesting that much lower levels can lead to quality loss
in sensitive fruits or vegetables (Wills et al., 1999, 2000). It is clear that this
molecule has a huge impact in conventional handling and distribution sys-
tems, and increased understanding of its biology and mechanisms of effect
on quality loss is essential to reducing loss during postharvest handling.
Ethylene is produced via the methionine (Yang) cycle (Figure 2.1)
in the cell wall–cell membrane complex and is only functional when the

5’-Methylthioribose-
Methylthioribose kinase 1-phosphate

α-Keto-γ-methylthio-
5’-Methylthioribose
butyric acid
Methylthioadenosin Methionine
nucleosidase Transaminase
Cycle
5'-Methylthioadenosin Methionine

Fruit ripening, auxin, wounding or ⊕ S-Adenosyl-

SAM synthetase
infection, chilling, water stress, anoxia ACC Synthase L-methionine
AVG, AOA − (SAM)
se
id a
ox

1-Amino-cyclopropane- CO2
C
AC

N-Malonyl-ACC (MACC) carboxylate Ethylene (C2H4)

yl- (ACC) −
on ⊕ −
CP
al 1-M
-m
N se
2,
len CO

C fera
Low emperat rs
high l scaveng
radic

C
A an s
thy high

tr Ethylene receptor
e

oxyg
t
a

sites
ou ing,

en, c re, free

gen en
se
exo it rip

obalt
u
e
Fru

Figure 2.1  Representation of the methionine (Yang) cycle showing the

control points in the cycle in regard to ethylene production. (Adapted from
Wang, K.L.-C. et al., The Plant Cell (Suppl.), S131–S151, 2002; Blankenship,
S.M., and Dole, J.M., Postharvest Biology and Technology 28, 1–25, 2003;
Saltveit, M.E., Postharvest Biology and Technology 15, 279–292, 1999.)

52
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

membrane is associated with the cell wall (Mattoo and Lieberman, 1977).
This understanding of the localization of production may explain the
observations that ethylene production is transient during ripening and
senescence or in response to wounding (Figure 2.2). The latter stages of
senescence involve cell wall breakdown and cell membrane disintegration
(Hurkman and Kennedy, 1975), which would imply that the association
of membrane with cell wall is lost. Also in wounding, cell membranes are
known to lose their functionality (Toivonen and DeEll, 2002), again imply-
ing the loss of cell wall membrane associations that exist in intact cells.
Transient ethylene production occurs only at the onset of climacteric ripen-
ing and senescence (Biale et al., 1954; Aharoni and Lieberman, 1979) or
early in the wound response (Toivonen and DeEll, 2002), which is consis-
tent with this hypothesis.
Fruits and vegetables have varying sensitivities to ethylene, and
responses to ethylene are manifested in many different ways. Table 2.1
shows the relative sensitivity of many fruits and vegetables commonly mar-
keted in North America. It is also important to note that many of the sensi-
tive fruits and vegetables can also produce significant amounts of ethylene,
because ethylene production is linked to the ripening of that fruit. Table 2.2
lists the responses that are evoked with exposure to ethylene by various

Climacteric peak
Carbon dioxide production

Post-climacteric
decline
Climacteric
rise

Pre-climacteric
minimum

Non-climacteric pattern

Time

Figure 2.2  Respiratory patterns for climacteric and nonclimacteric fruit.

(From Saltveit, M.E., in Gross, K.C. et al. (eds.), The Commercial Storage
of Fruits, Vegetables, and Florist and Nursery, USDA Handbook 66, U.S.
Department of Agriculture, Agricultural Research Service, Beltsville, MD,
2004, http://www.ba.ars.usda.gov/hb66/019respiration.pdf.)

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 2.1  Relative Production Rates and Sensitivity to Ethylene in Selected

Fruits and Vegetables
Relative Ethylene Relative Sensitivity to
Fruit or Vegetable Production Rate Ethylene
Apple Very high High
Globe artichoke Very low Low
Asparagus Very low Moderate
Avocado High High
Banana Moderate Moderate
Green bean Low Moderate
Raspberry Low Low
Strawberry Low Low
Broccoli Very low High
Cabbage Very low Moderate to high
Carrots Very low High
Cauliflower Very low High
Celery Very Low Moderate
Cherries Very low Low
Cilantro Very low High
Citrus fruit Very low Moderate
Corn Very low Low
Cucumber Low High
Eggplant Low Moderate
Garlic Very low Low
Grape Very low Low
Chives Low Moderate
Kiwifruit Low High
Leafy greens Very low High
Lettuce Very low High
Mango Moderate Moderate
Mushrooms Very low Moderate
Mature onions Very low Low
Nectarines, plums, and peaches Moderate Moderate
European pear High High
Peppers Low Low
Pineapple Low Low
Potato Very low Medium
Squash Low Medium

(Continued )

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Table 2.1  Relative Production Rates and Sensitivity to Ethylene in Selected

Fruits and Vegetables
Relative Ethylene Relative Sensitivity to
Fruit or Vegetable Production Rate Ethylene
Sweet potato and yam Very low Low
Tomato, mature green Very low High
Tomato, firm ripe High Low
Watermelon Very low High
Source: Extracted from Cantwell, M., in Kader, A.A. (ed.), Postharvest Technology
of Horticultural Crops, Agriculture and Natural Resources Publication
3311, University of California, Oakland, 2002, pp. 511–518.

Table 2.2  Changes or Disorders Caused by Ethylene Exposure for Selected

Fruits and Vegetables
Fruits or Vegetables Affected Response to Ethylene Exposure
Apple, avocado, pear, stone Accelerated climacteric ripening, softening
fruits, mature green tomato,
melons, watermelon, banana
Carrot, parsnip, celery Production of phytoalexins (iso- or
furanocoumarins) leading to bitterness;
furanocoumarins can cause contact
dermatitis on exposure to UV light when
handling vegetables with high contents
Leafy vegetables, cruciferous top Premature yellowing and senescence
vegetables, green asparagus,
herbs, citrus, cucumbers
Iceberg lettuce Postharvest foliar russet spotting
Asparagus Toughening through induction of lignification
Potatoes, bulb onions Sprouting at high concentrations
Potatoes Inhibition of sprouting at low concentrations
Mushrooms Browning of caps
Sweet potatoes Hard core (inedible hardening of flesh
evidenced upon cooking)
Sources: Information compiled from Timbie, M., and Haard, N.F. et al., Journal
of Food Science 42, 491–493, 1977; Gross, K.C. et al., The Commercial
Storage of Fruits, Vegetables, and Florist and Nursery Stocks, USDA
Handbook 66, U.S. Department of Agriculture, Agricultural Research Service,
Beltsville, MD, 2004, http://www.ba.ars.usda.gov/hb66/; Toivonen,
P.M.A., in Hui, Y.H. et al. (eds.), Handbook of Vegetables and Vegetable
Processing, Wiley-Blackwell Publishing, Ames, IA, 2010, pp. 199–220.

55
P OST H AR V EST RI P ENING P H Y SIOLOG Y

groups of fruits and vegetables. A part of produce incompatibility in stor-

age is based on the impact of ethylene on fruits and vegetables that do not
normally produce significant levels of ethylene but are sensitive to ethylene
exposure.

2.2.2  Respiratory Characteristics:

Climacteric and Nonclimacteric
The respiratory behavior of fruit during ripening can be classified as
­climacteric or nonclimacteric in nature. In nonclimacteric fruit, the res-
piration rate declines slowly after harvesting. This respiratory decline is
generally assumed to occur as a consequence of depletion in substrates
available for respiration. In contrast, climacteric fruit exhibit rapid and dra-
matic respiratory increase during ripening (Figure 2.2). The climacteric
rise has been characterized as having four distinct phases, the first being
a transient decline in respiration rate at full fruit maturity, followed by a
rapid rise that reaches a maximum and then declines as the fruit tissue
deteriorates. In general, the climacteric peak is normally many orders of
magnitude above basal preclimacteric respiration.
The respiratory behavior has implications for the storage and
­handling of fruits or fruit-vegetables. In general, nonclimacteric fruits or
fruit-vegetables do not exhibit accelerated ripening in response to ethyl-
ene exposure; however, they may show other ethylene responses, such as
­yellowing, secondary development, or secondary compound accumulation
(Saltveit, 1999).

2.2.3  Developmental or Maturity Stage at Harvest

Harvest maturity is an important subject, and much effort is placed in
selecting the optimal maturity stage for all fruits and vegetables to ensure
the best quality and storage potential. Watada et al. (1984) defined the
developmental stages for horticultural crops in great detail (Table 2.3).
The physiological implications of developmental stage and the degree of
maturity at that stage are tremendous, and a misunderstanding could lead
to poor decisions in postharvest handling protocols. For example, tomato
fruit must be at “mature green” when internally the seeds are tan in color
and there is a significant development of a gel in at least two locules of the
fruit (Sargent and Moretti, 2004). If the fruit has not reached this stage
of maturity at harvest, they will not ripen, even if exogenous ethylene is
applied (Sargent and Moretti, 2004). However, generally the identification
of mature green can be difficult in practice, and the commercial industry

56
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Table 2.3  Classification of Fruits and Vegetables Based on Developmental

Phase and Anatomical Structure
Anatomical
Phase in Development Description of Fruit
Continuum or Vegetable Examples
Vegetative growth Sprouts Alfalfa, onion, watercress,
bean sprouts
Stems or leaves Asparagus, celery, spinach,
lettuce, cabbage, kohlrabi,
chard, endive, herbs
Inflorescences Artichokes, broccoli,
cauliflower

Late vegetative growth Partially developed Cucumber, green beans,

to physiological fruit okra, sweet corn, green
maturation peppers, summer squash

Ripening Fully developed fruit Apple, pear, citrus, tomato,

colored peppers, mango,
papaya, avocado, stone
fruit, melons, winter squash
Roots and tubers Carrot, onion, potato, garlic,
Jerusalem artichoke,
jicama, ginger
Source: Adapted from Watada, A.E. et al., HortScience 119, 20–21, 1984.

often harvests at a “breaker” stage to ensure that the fruit will respond to
ethylene and ripen after harvest.
The rate of developmental change is also important to understand in
handling fruits and vegetables. In general, those fruits and vegetables har-
vested at the vegetative growth stage to the physiological maturity stage
of the development continuum will have the most rapid metabolism and
hence shortest shelf potential (Toivonen, 2010). In addition, there is usu-
ally a higher respiratory rate, and it is well known that shelf life is consid-
ered to be inversely proportional to respiration rate (Saltveit, 2004). For
fruits and vegetables harvested at the ripening phase of the developmental
continuum, if the correct timing is chosen, respiration rates can be quite
low; for example, if potatoes are allowed to grow to maturity and are cured
before ­storage, they have much lower respiration rates than if they are
harvested as immature potatoes (Toivonen, 2010). Also, climacteric fruit,
which can exhibit high respiration rates in the latter stages of ripening,

57
P OST H AR V EST RI P ENING P H Y SIOLOG Y

when harvested at the early climacteric stage and properly stored, have
relatively low respiration rates and can be stored for long periods of time
(Saltveit, 2004; Watkins et al., 2004).
Because timing of harvest is an important factor in determining
storage life, various practical indices have been developed to aid in mak-
ing optimal harvest decisions, ranging from visual and tactile indices to
precise instrumental measures (Reid, 2002). The approach in each case
depends on monitoring visual, tactile, or compositional changes that occur
in the fruit or vegetable, leading to the horticultural optimal maturity for
­harvest. In the case of lettuce, this means assessing the solidity of the head;
in apples, this means measuring the loss of starch with iodine staining,
and in persimmons the tannin content (Reid, 2002). Clearly, each fruit or
vegetable has a distinct approach to assessing maturity for harvest. In addi-
tion, there continues to be challenges for fruits and vegetables or new culti-
vars emerging into the export markets, where practical and reliable harvest
maturity indices have yet to be developed.

2.2.4  Tolerance to Low Temperatures

Sensitivity to chilling-induced quality changes or tissue injuries is an
important issue for postharvest handling and storage of subtropical and
tropical fruits and vegetables, but it can also be of concern in the handling
and storage of temperate fruits and vegetables (Wang, 2004). The thresh-
old temperatures for chilling injury can range from 13°C down to 0°C;
however, the mechanism for chilling-related disorders involves changes
in membrane functionality (Wang, 1982). Secondary events stemming
from loss in membrane functionality lead to chilling-associated disor-
ders, which can include internal browning, soft scald, skin darkening or
discoloration, abnormal ripening or loss of ability to ripen, tissue necro-
sis, pitting, hardening, internal discolorations, loss of aroma production,
and decay (Figure 2.3). It should be noted that decay often is secondary
to physical injury induced by the chilling response (Morris, 1982).
There has been much research devoted to modulating or eliminating
chilling-induced injury (Wang, 1993). Many of the proposed approaches to
alleviating chilling injury involve mechanisms that cause the modification
of membrane lipid composition, allowing the membrane to sustain func-
tionality (Luengwilai et al., 2012). Other approaches involve modulating the
metabolism of the fruit or vegetable such that chilling-induced changes are
minimized (Wang and Qi, 1997). It is also known that the more mature a
fruit is, the less susceptible it is to chilling injury, presumably because more
mature fruit has a higher enzymatic or nonenzymatic antioxidant content,
which helps to protect membranes from oxidative injuries (Qian et al., 2013).

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Chilling temperature

Chilling sensitive fruit

or vegetable

Primary response
Phase transition of membrane
from liquid crystalline to solid gel

Secondary responses
ROS accumulations
Ethylene production
Increased respiration
Decoupling of electron transport from
energy production
Alteration of cellular structure
Alterations in metabolic pathways

Recovery
Short exposure
to pre-stressed
–processes reversible
functionality

Prolonged exposure

Symptoms of injury
Discoloration
Surface pitting
Internal breakdown
Loss of capacity to ripen
Wilting
Decay
Aroma Loss

Figure 2.3  Schematic representation of primary and secondary c­hilling

responses and ensuing symptoms that develop should the exposure
be prolonged. (Adapted from Wang, C.Y., HortScience 17, 173–186,
1982; Sevillano, L. et al., Journal of the Science of Food and Agriculture 89,
555–573, 2009.)

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

More recently, efforts have been focused on directed breeding to

confer chilling resistance in new cultivars of fruits in particular (Martínez-
García et al., 2012; Dhanapal et al., 2012). While genetic markers for chill-
ing resistance are being developed, the trait is not straightforward, and
the linkage to specific gene regulation is not well understood at this time
(Martínez-García et al., 2012). This problem is likely due to the fact that,
physiologically, the expression of chilling symptoms used to develop genetic
markers may involve secondary processes (Wang, 1982), and so a clear rela-
tionship between chilling symptoms and the primary processes leading to
them is not easily defined. Nevertheless, continued efforts to use genetic
markers to increase chilling resistance is a worthwhile effort that will
allow identification of chilling-resistant germplasm from strategic crosses
between resistant and commercially desirable parents. Because of the com-
plex nature of the gene expression relationship to chilling symptoms, it
is likely that genetic engineering will not provide significant efficiency at
achieving commercially desirable fruit with chilling tolerance (Vinocur and
Altman, 2005).

2.3  Factors Affecting Physiology of Fruits

and Vegetables in Postharvest Systems

2.3.1 Temperature
Temperature influences the rate of all metabolic processes, including res-
piration. Metabolic processes all rely on enzymatic activity, which can be
further generalized as a chemical reaction. As such, effects of temperature
can be described by an Arrhenius plot, which is derived as
An enzymatic reaction can be generalized as

Substrate A + Substrate B → Product A + Product B

The rate of the reaction is defined as

V = k × [Substrate A] × [Substrate B]

where V is the velocity of the reaction and k is the velocity constant.

The constant k is determined from an Arrhenius plot, where
µ
−
k = Ae RT

where A is a constant, µ is the activation energy, R is the perfect gas
­constant, and T is the absolute temperature.

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

The effect of temperature change on reaction rate is defined as the

quotient of the rate of change over a 10°C change in temperature (i.e., Q10).
The Q10 for a particular reaction or enzyme activity is calculated as

Vt +10
Q10 =
Vt

where Vt is the rate measured at a temperature (t) measured in degrees
Celsius and Vt+10 is the rate at 10°C higher than t.
The Q10 has been widely used in the context of respiratory behavior
cold storage and to explain the effect of low temperatures on enhancing
the storage life of produce (Nunes and Emond, 2003; Bron et al., 2005).
However, it has to be noted that not all enzymes involved in respiratory
and nonrespiratory pathways have identical Q10 values in the tempera-
ture ranges studied (0°C–20°C). Pollock and ap Rees (1975a) demon-
strated that enzymes involved with sugar metabolism have wide ranges in
Q10 ­values, between 2°C and 10°C, and this consequently explained some
of the changes associated with low temperature sweetening in potatoes
(Pollock and ap Rees, 1975b). It has also been shown that enzyme systems
can become acclimatized to low temperatures, becoming less sensitive
to temperature change over time, and thus having lower Q10 values after
time in storage (Atkin and Tjoelker, 2003). Rhodes et al. (1981) demon-
strated differential changes in enzymes involved with phenolic metabo-
lism in response to temperature and differential temporal changes in
these enzyme activities with time at low temperature storage. This work
clearly emphasizes the importance of temperature for imposing shifts in
metabolic pathways, and doing so in such a manner that relative contents
of phenolic constituents are modified, and this relationship also shifts
over time as some of the enzymes acclimate, and others do not, to low
temperatures.
While Q10 principles have been applied to enzymatic processes in
fruits and vegetables, temperature also has effects on physical move-
ments into, out of, and within fruit tissue; that is, rates of gaseous and
liquid diffusion in tissues can be influenced by temperature (Lammertyn
et al., 2001). This finding emphasizes how little is known about the com-
ponents of apparent Q10 values developed for postharvest analysis (Bron
et al., 2005).
The importance of low temperatures to controlling quality decline in
fruits and vegetables is well known (Nunes and Emond, 2003). However, a
scan of the literature indicates that there are limited reports on differen-
tial effects of low temperature on fruit and vegetable synthetic, catabolic,
and respiratory pathways. The lack of good tools to measure a broad range
of proteins (enzymes) and metabolic substrates and products has likely
­limited work in this area. However, with new tools and data processing

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capability for proteomic and metabolomic analysis, the investigation of dif-

ferential changes in a broad range of enzymes and metabolites in metabolic
pathways has now become a reality (Pedreschi et al., 2009; Rudell et  al.,
2009). Hopefully this will lead to a better understanding of the effects of
storage regimes, including temperature, on respiratory, metabolic, and
quality changes in fruits and vegetables.

2.3.2 Humidity
Humidity is the second most important factor, after temperature, for mod-
ulating changes in physiology and the quality of fruits and vegetables in
postharvest systems. The humidity of the air surrounding a fruit or veg-
etable exerts some vapor demand on the produce if the humidity is below
100%, which is normally the case in postharvest systems. Water loss can be
­calculated as

J=
(P − P ) × A
i a f

( RD × T ) × r

where J is the water loss in weight of water per unit time per surface area
of the fruit, Pi and Pa are the steady-state vapor pressures of water in the
intercellular spaces of the fruit and in the atmosphere, respectively, A f is
the surface area of the fruit, R D is the gas constant, T is the absolute tem-
perature in degrees Kelvin, and r is the specific resistance of the fruit to
water loss. While this model provides precise estimates of water loss, it is
essential to determine the surface area of the fruit and the specific resis-
tance of that fruit to water loss. In practice, it is more important to get an
estimate of the Pi − Pa component of the water loss model so that relative
changes in driving force for water loss (i.e., vapor pressure deficit) can
be quantified, allowing evaluation of the effect of changes in postharvest
handling protocols on relative water loss from the fruit.
In order to calculate vapor pressure deficit, the saturation vapor pres-
sure of the fruit or vegetable and the surrounding air must be calculated.
Saturation vapor pressure (VPs ) can be calculated as
17.27 × T
VPs = 0.6108 × e T + 237.3

where T is the temperature of the produce or the surrounding air in degrees
Celsius.
The intercellular spaces of the fruit or vegetable are considered to be
at saturation vapor pressure (Ben-Yehoshua and Rodov, 2003). However,
the air vapor pressure (VPa ) can be calculated as

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

RH a
VPa = × VPsa
100

where RHa is the relative humidity of the air in percent and VPsa is the calcu-
lated saturation vapor pressure of the air at that temperature.
Water vapor deficit (VPD) is the driving force for water loss from
fruits or vegetables and is calculated as

VPD = VPp − VPa

where VPp is the calculated saturation vapor pressure of the fruit or veg-
etable and VPa is the calculated vapor pressure of the surrounding air based
on its temperature and relative humidity.
It is a straightforward task to calculate vapor pressure deficits for
many postharvest handling scenarios to demonstrate the need for rapid pre-
cooling compared to room cooling. It can be demonstrated that warm fruit
will have higher integral vapor pressure deficits if room cooled, and hence
greater water loss than fruit that is rapidly forced-air cooled (Toivonen
and Hodges, 2011). Clearly, it is important to cool fruits and vegetables to
temperatures as close to that of the storage room or transport container as
soon as possible in order to minimize water loss. In addition, it is impor-
tant to manage the relative humidity in a storage room or container such
that it is maintained as high as is suitable for a particular fruit or vegetable
(Cantwell, 2002).
As implied earlier, there are a number of factors that determine the
resistance of produce to water loss—the composition and thickness of
the epidermis; the presence of surface structures on the epidermis, such
as hairs; the number or presence of stomata (or lenticels); and the mor-
phological structure (a bulky organ with a low surface area-to-volume
ratio vs. a leafy green with a high surface area-to-volume ratio). The
resistance to water loss can be modulated with wax coatings or applica-
tion of packaging to control humidity around the produce (Toivonen and
Hodges, 2011).
If water loss is not controlled, there is certainly a loss of turgor leading
to wilting, but there are also a few other metabolic changes that can occur
(Toivonen and Hodges, 2011). The enzyme activities of polygalacturonase
and pectinesterase increase, leading to loss of cell wall structure and con-
comitant increases in soluble sugars in tomato (Inari et al., 2002). Similar
results have been found with cucumbers, in which water stress caused an
upregulation of polygalacturonase activity, leading to the hypothesis that
the physical loss of water (i.e., loss of turgor in the tissue) was not the only
factor in causing softening of stressed fruit (Kubo et al., 2000). Water stress
also induces increased ethylene production (Kubo et al., 2000), and this

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may explain accelerated ripening in water-stressed bananas (Burdon et al.,

1994) and accelerated senescence in bell peppers (Lurie et al., 1986).

2.3.3  Atmospheric Modification

Modified and controlled atmospheres have been long used to modulate
fruit and vegetable physiology and metabolism in order to extend their stor-
age lives (Kader and Saltveit, 2003a). A wide range of physiological effects
on fruits and vegetables in response to reduced O2 or elevated CO2 have
been documented (Kader and Saltveit, 2003a). While the prior review of
this subject area is complete and up to date, a few points are worth not-
ing in this discussion. In many cases, both reduced O2 and elevated CO2
result in inhibition of many pathways or enzyme activities; however, in
some cases (e.g., phenylalanine ammonia lyase [PAL] activity, succinic
acid and malic acid accumulation, and chilling injury) they may counteract
each other. Therefore, overall quality change may be difficult to manage
using controlled and modified atmospheres, since some pathways respond
­differently. In one case, it has been shown that a high CO2 atmosphere
improves overall quality retention of stored strawberries; however, malic
acid content losses increase and lead to potential flavor loss since malic acid
is an important aspect of fruit flavor (Holcroft and Kader, 1999).
While controlled and modified atmosphere protocols have been devel-
oped to optimize storage life of fruits and vegetables, there are risks of using
the technology in certain cases (Kader and Saltveit, 2003). These cases
include aggravation of disorders such as blackheart in potatoes, irregular
ripening of melons and tomatoes (when O2 < 2% and CO2 > 5%), develop-
ment of off-flavors (when O2 < 0.5% and CO2 > 20%), increased susceptibil-
ity to decay in produce that has been injured in extreme atmospheres, and
simulation of sprouting and inhibition of periderm development in root and
tuber vegetables. Therefore, it is important to understand the physiologi-
cal parameters of each fruit and vegetable and the risk–benefit relationship
for atmosphere modification. Atmosphere recommendations for most fruits
and vegetables are listed by Cantwell (2002).
An emerging area of interest is the use of dynamically controlled
atmosphere (DCA) storage, which in principle works through adjusting
atmospheres during storage to meet changes in fruit acclimation to stor-
age conditions (Kader and Saltveit, 2003a). The challenge for this new
approach is to have monitoring instrumentation that can sense fine physi-
ological changes in the fruit, enabling appropriate atmosphere adjustment
while avoiding tissue injury (Saltveit, 2003). It also must be noted that this
technology, like most, may not be beneficial for all cultivars, and there
are significant new considerations that must be met before successful

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

implementation can take place in commercial practice (Watkins, 2008).

There is much need for further research to better understand the effect
of dynamically controlled ultralow oxygen on fruit physiology and
metabolism.

2.3.4  Abiotic Stresses

Postharvest handling by definition imposes abiotic stress on a fruit or
vegetable (Toivonen and Hodges, 2011). However, the fruit or vegetable
may be subject to preharvest factors or stresses that also lead to modified
stress susceptibility in postharvest handling and storage (Toivonen, 2003;
Toivonen and Hodges, 2011). In general, three possible scenarios can exist
when a fruit or vegetable has been harvested: (1) it can be in an unstressed
state and, as such, can withstand many stresses imposed by postharvest
processes; (2) it can have been subjected to mild stresses, which enhance
its resistance to stresses imposed by postharvest processes; or (3) it can
have been subjected to severe stresses, which weaken its resistance to sub-
sequent stress exposure in postharvest processes.
Ripeness stage, storage temperature, ethylene, and oxidative stress
all affect peel phytosterol metabolism in apples and thus influence the
development of superficial scald (Rudell et al., 2011). There has been some
work showing that sun exposure on the tree may lead to invisible dam-
age on apple fruit (Kuckenberg et al., 2008), which may contribute to the
de­velopment of postharvest scald (Hernandez et al., 2014). These observa-
tions suggest that scald is associated with chilling responses. When review-
ing the approaches being adopted to control disorders such as superficial
scald in apples (Rudell et al., 2011), it becomes clear that these are the same
approaches that have been adopted to modulate chilling injury in other
fruits and vegetables (Sevillano et al., 2009).
Mechanical injury is a consequence of physical handling procedures
employed during the harvest and packing of fruit. If often leads to bruis-
ing in fruit such as apple, and the visual appearance of the bruising can be
reduced by an “apparent recovery” or suberization period after the bruising
injury is incurred (Toivonen et al., 2007). Proteomic analysis of response
to mechanical injury in apples has shown that response to stress proteins,
many of which are considered to be pathogen resistance–related proteins,
was upregulated with wounding (Buron-Moles et al., 2014). This supports
the overall hypothesis of cross-tolerance mechanisms in fruit and vegeta-
ble tissues, where it is suggested that the responses to biotic and abiotic
stresses share many common gene and metabolome expression pathways
(Toivonen, 2004).

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2.4  Physiological Changes Occurring during

Postharvest Handling or Storage

2.4.1  Depletion of Respiratory Substrate

Despite attempts to inhibit respiration in postharvest systems with the
use of low temperatures, controlled or modified atmospheres (Kader and
Saltveit, 2003b), and other technologies, such as 1-methylcyclopropene
(Blankenship and Dole, 2003), there is an inevitable loss of respiratory
substrate over time, which can be significant with long storage durations
extending several months. The consequence of such depletion is clearly
associated with important quality attributes of fruits and vegetables.
Loss of malic acid content is a significant problem in the long-term stor-
age of some fruits, such as apple, where malic acid is an important aspect
of sensory quality (Jan and Rab, 2012). While the sugar concentration
of apples generally increases during long-term storage, the dry matter
content declines (Jan and Rab, 2012). Dry matter has also been recently
identified as an indicator of poststorage quality in apples (Palmer et al.,
2010). It is clear from these observations that dry matter and respiratory
organic acids such as malic acid are depleted due to respiratory activity
over long-term, low-temperature storage, whereas sugar concentrations
are maintained or increased. It may be that low-temperature effects on
sugar metabolic enzymes (ap Rees et al., 1981) are partially responsible
for this apparently contradictory response. Dry matter content at harvest
has also been shown to be important for the short-term storage of broccoli
microgreens (Kou et al., 2014), suggesting that the relationship between
dry matter and sustained respiration during storage holds true for fruits
and vegetables.
The potential storage life of fruits and vegetables is partially deter-
mined by their relative respiration rates; however, the other side of the issue
is that storage life comes to an end when a critical threshold for a respira-
tory substrate has been reached (Kader and Saltveit, 2003b). Respiration
is essential for the living tissues to maintain cellular organization and
function through the maintenance of adequate adenosine triphosphate
(ATP) titers in the cells, allowing the ongoing basal metabolic activities
to continue. As respiratory substrates are depleted, the ability to regener-
ate ATP declines, which could potentially affect fruit tissues and quality
retention. There has been some work done to demonstrate the importance
of the energy state on the postharvest development of quality defects in
apple and pear fruit (Saquet et al., 2000). More recently, it has been sug-
gested that regulation of the energy state in fruit may be the key to manag-
ing ripening and senescence in storage (Wang et al., 2013). The concept

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

of energy homeostasis has emerged from this work (Figure  2.4), which
is congruent with earlier understanding in regard to the maintenance of
cellular organization and function. Clearly there is significant understand-
ing yet to be had in regard to the maintenance metabolism of tissues in
storage.

Stress (senescence)

+ −

Sucrose
ABA ATP Deficit

SnRK Signaling
Pathway

ATP Transporting ATP Synthesising ATP Dissipating Energy homeostasis

(AAC/ANT) (AtpB) (AOX/UCP)

ROS
Fruit Quality
Deterioration

Figure 2.4  Possible mechanism to account for energy regulation in senescent

litchi fruit. An energy deficit in intensive respiration during the senescence pro-
cess may be sensed by sucrose non-fermenting-1-related kinase (SnRK), which
controls the expression and phosphorylation of key metabolic enzymes, and
this might involve ATP synthase, ADP/ATP carrier (AAC), alternative oxidase
(AOX), and uncoupling mitochondrial protein (UCP). The energy homeostatic
condition can be maintained to a certain extent, but the equilibrium is ulti-
mately disrupted, which is correlated with fruit deterioration. (From Wang, H.
et al., BMC Plant Biology 13, 55, 2013.)

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2.4.2  Hormonal Effects

There are two aspects of hormonally associated changes in postharvest
systems. The first is related to endogenous shifts in hormone levels and
the consequent metabolic changes, and the second involves the effects of
exogenously applied hormones. Early work focused on measures of endog-
enous hormones and developmental changes in fruit and vegetable tissues
(Ludford, 2003). Work on exogenously applied phytohormones followed
that research (Ludford, 2003; Klein and Goldshmidt, 2005) and continues
to present. However, only recently have advances in this area occurred on
a broader genomic, proteomic, and metabolomic scale. Genomic studies
evaluating the response of fruit to endogenous hormone contents have pro-
vided a broader, more integrated understanding of response mechanisms
(Tacken et al., 2010; Devoghalaere et al., 2012; Matsuo et al., 2012). This
broader and more integrated analysis of responses is already better defin-
ing the complex metabolic changes that are induced (Pech et al., 2008),
and this is expected to lead to a more useful and reliable understanding of
control mechanisms.
Ethylene is probably the most studied hormone in postharvest research
literature. It is associated with the onset of ripening and senescence (refer
to discussion in Section 2.2.1), and as such, it has been employed to acceler-
ate or initiate ripening in fruits that are harvested green. Ethylene is rou-
tinely used for ripening green bananas and tomatoes, degreening lemons,
and conditioning pears that are harvested green (Toivonen, 2010; Sugar and
Basile, 2013). It has been discovered that higher levels of ethylene expo-
sure can be applied to inhibit sprouting in potato tubers (Daniels-Lake
et al., 2005). While ripening in climacteric fruit is considered to be highly
associated with ethylene, it is clear that changes at the molecular level are
­complex, involving both the gene transcription level (Nishiyama et  al.,
2007) and the posttranscriptional level, where micro-RNAs may be oper-
ating to modulate gene expression through differential silencing of target
genes involved with ethylene biosynthesis and signaling (Zuo et al., 2012).
Also, it is becoming clear that there is ethylene-dependent and -­independent
regulation of metabolic pathways associated with ripening (Zuo et al., 2012),
the impact of which is yet to be fully understood.
Gibberellic acid (GA) has been used for preharvest management of
fruit quality in many fruits. It can increase citrus size and reduce seed set in
facultatively parthenocarpic citrus cultivars (Iglesias et al., 2007). In sweet
cherries, GA has been adopted widely in commercial practice to delay
fruit maturation and increase fruit size and firmness (Canli and Orhan,
2009). Similar responses have been found in seedless table grapes with
GA applied at times soon after fruit set (Singh et al., 1978). The basis for
these responses is thought to be that endogenous GA normally produced

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

by the seed is either replaced or enhanced with the exogenous application.

Exogenous GA application has been shown to induce increases in fruit
length and girth, as well as soluble solids contents (Banerjee and Basu,
1992). GA has also been shown to downregulate polygalacturonase in sweet
cherries, which may provide some insight into its effect on fruit firmness
(Choi et al., 2002). Other work, with persimmon, has shown that exogenous
GA application results in modulation of expression for an expansin gene
(Zhang et al., 2012). Expansin is associated with fruit size or advance in
ripening of fruit (Brummell et al., 1999; Hiwasa et al., 2003), which may
explain the increased fruit size and delay in ripening of sweet cherries
treated with exogenous GA (Canli and Orhan, 2009). Also, higher levels of
production of endogenous GA early in development in Japanese pears have
been shown to relate to larger fruit size in one cultivar when compared to
one that produces very low levels (Zhang et al., 2007). In relation to matu-
rity, exogenous GA application has been shown to reduce sugar supply to
orange peel, suggesting that a delay of peel maturation could be directly
due to limitation in sugar substrate to that tissue (Fidelibus et al., 2008).
Clearly, a more in-depth and integrated analysis of GA effects on gene
expression and metabolome is necessary to better understand the varied
physiological effects of this hormone, especially since it is used extensively
in the commercial horticulture to modulate fruit quality.
Abscisic acid (ABA) has been shown to have a range of effects when
applied to different fruits. In southern highbush blueberry, ABA application
resulted in delayed maturity and firmer fruit, but had no other quality or
nutritional effects (Buran et al., 2012). In contrast, exogenous ABA enhanced
antioxidant capacities, anthocyanins, and flavonol contents in muscadine
grapes (Sandhu et al., 2011), although not all cultivars tested showed a sig-
nificant response. Cantín et al. (2007) found that exogenous ABA treatment
improved color development and visual appearance of Crimson seedless
grapes in comparison to industry existing practices. Jiang and Joyce (2003)
found that exogenous ABA accelerated softening and color development in
strawberries, and this was linked directly to effects of ABA on ethylene syn-
thesis and phenylalanine ammonia lyase upregulation, respectively. High
levels of ABA production have been linked to small fruit size in Japanese
pear (Zhang et al., 2007). Sun et al. (2013) recently investigated gene expres-
sion in melon fruit development and ripening, finding that genes encoding
non-ethylene-dependent ripening changes were regulated by ABA, and this
includes the induction of climacteric ethylene production. They suggested
that ABA plays a significant role in the early stages of ripening in the fruit,
and that ethylene takes over the major regulatory role at later stages.
Exogenous auxin application has been shown to increase fruit size,
enhance color, and advance ripening in the Maxim® apricot (Bregoli
et al., 2010). In a more critical study, a large number of genes have been

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shown to be either repressed or induced by exogenous auxin application in

­strawberry (Aharoni et al., 2002). In general, the auxin-related changes in
gene expression were linked to cell wall metabolism and stress response
(including ethylene production). In a more detailed study, auxin response
was found to correlate to a quantitative trait loci (QTL) associated with fruit
weight in apples (Devoghalaere et al., 2012). Their results also suggest that
exogenous application of auxin at low concentrations (10 −7 M indoleacetic
acid) in early fruit development could enhance apple diameter; however,
higher concentrations (10 −5 M indoleacetic acid or higher) could negate
the size increase response or even reduce fruit size relative to untreated
controls. Response to auxin is very much determined by the timing of exog-
enous application and the concentration that is applied. However, in the end,
it seems that modulating auxin response is best managed through QTL
analysis and directed breeding approaches since factors affecting exog-
enous application may not be easily managed in a wide range of cultivars,
and the auxin response is extremely complex.
Exogenous cytokinin applications have been shown to delay loss of
chlorophyll and senescence in green leafy vegetables (Ludford et al., 2003).
This response has been linked to upregulation of genes for light-­harvesting
cytokinin-binding proteins and stimulation of chloroplast-encoded tran-
scription in vegetative tissues (Kulaeva et al., 2000; Zubo et al., 2008).
However, more recently it has been demonstrated that increases in endog-
enous cytokinin levels in kiwifruit lead to effects on response regulators
to the STAY-GREEN2 gene (Pilkington et al., 2013). This work suggests
that cytokinin is important to chlorophyll contents of green developing
­k iwifruit; however, it does not show that cytokinin is responsible in control-
ling the ripening-associated loss of chlorophyll in the fruit. In tomato fruit,
cytokinin has been demonstrated to promote the activity of genes involved
with cell division during early fruit development (Matsuo et al., 2012).
Phytohormones exert profound regulatory control over fruit and
vegetable development and final quality. The preceding discussion has
shown that the application of recent advances in metabolomic, proteomic,
and genomic analyses has created a greater understanding in hormone
effects on different aspects of fruit growth, development, and ripening.
However, endogenous phytohormones coexist in the intact fruit and vary
in level temporally during fruit development and ripening, and may modu-
late or even counteract each other’s activities in some cases. For example,
auxin and abscisic acid behave antagonistically in the expression of a
SHAT TERPROOF-like gene in the ripening of strawberry (Daminato et al.,
2013). The SHAT TERPROOF-like gene has a profound effect on modulat-
ing the expression of many genes regulating the ripening of the strawberry.
In another example, it has been demonstrated that exogenous cytokinin-
induced parthenocarpy in tomato was partially due to the effects on cyto-
kinin in modulating GA and auxin metabolism (Ding et al., 2013). This last

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P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

report highlights the importance of the coordinated regulatory actions of

all phytohormones in effecting the development, growth, and ripening of
fruit. The understanding at the genome and metabolome level is only now
beginning to develop and in the future should explain the physiological
effects that have been documented in earlier literature.

2.4.3  Membrane Alterations

As mentioned above in the discussion surrounding chilling stress, altera-
tion of membrane function is the primary response to that stress. Chilling
and other stresses alter the function of membrane proteins and synthetic
pathways, leading to many changes in metabolite accumulations, includ-
ing o­ xygen free radicals (Alcázar et al., 2010). Consequent tertiary effects
include significant alterations in gene expression and loss of membrane
permeability and transport functions. Production of polyamines, which
is closely associated metabolically with ethylene synthesis (Alcázar
et al., 2010), has been shown to modify lipid bilayer membrane structure
and function such that increased tolerance to stress is realized. Other work
with apples has shown that storage stress leads to significant changes
in peel phytosterol metabolism (Rudell et al., 2011). Phytosterols are
­important to  membrane fluidity and function. Rudell et al. (2011) found
that steryl-6-O-fattyacyl β-D-glycoside accumulation in apple peel was an
indicator that the apples had suffered stress that could lead to superficial
scald injury of the peel. Others have observed oxidative injury membranes
in association with storage disorder development (Whitaker, 2004).

2.5  Conclusions and Future Perspectives

The physiology of harvested fruits and vegetables is very sensitive to condi-
tions of handling, and their sensitivity to handling can be modified by treat-
ments prior to harvest and after harvest. In the end, physiological sensitivity
and response are governed by gene, proteome, and metabolome expression
in the fruit or vegetable tissues. The research in postharvest physiology has
enjoyed many decades of advances using limited analysis techniques and rela-
tively easily measured parameters to judge physiological change in response
to handling and storage conditions. However, as new data are emerging from
talented research groups and networks, it is becoming clear that nuances
and unexpected outcomes noted in prior postharvest studies can potentially
be better understood by integrated use of emerging techniques in genom-
ics, proteomics, and metabolomics. The challenge of such approaches is that
they are quite costly and are more realistically supported through leveraged
funding and effort from a number of institutions and countries. Research

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groups providing the most comprehensive results are those who have truly
multidisciplinary collaborators, including crop production physiologists,
postharvest physiologists, chemists, and molecular b ­ iologists. This new
era of postharvest physiology requires new ways of working together and a
multidisciplinary approach to provide both perspective on the physiological
problem and a detailed understanding of the ­molecular mechanisms that
will allow us to better predict postharvest quality outcomes from treatment
applications and for directed breeding programs.

References
Aharoni, N., and Lieberman, M. 1979. Patterns of ethylene production in
senescing leaves. Plant Physiology 64, 796–800.
Aharoni, A., Keizer, L.C.P, Van Den Broeck, H.C., Blanco-Portales, R., Muñoz-
Blanco, J., Bois, G., Smit, P., De Vos, R.C.H., and O’Connell, A.P. 2002.
Novel insight into vascular, stress, and auxin-dependent and -independent
gene expression programs in strawberry, a non-climacteric fruit. Plant
Physiology 129, 1019–1031.
Alcázar, R., Altabella, T., Marco, F., Bortolotti, C., Reymond, M., Koncz, C.,
Carrasco, P., and Tiburcio, A.F. 2010. Polyamines: Molecules with regu-
latory functions in plant abiotic stress tolerance. Planta 231, 1237–1249.
ap Rees, T., Dixon, W.L., Pollock, C.J., and Franks, F. 1981. Low temperature
sweetening of higher plants. In Friend, J., and Rhodes, M.J.C. (eds.),
Recent Advances in the Biochemistry of Fruits and Vegetables. Academic
Press, London, pp. 41–61.
Atkin, O.K., and Tjoelker, M.G. 2003. Thermal acclimation and the dynamic
response of plant respiration to temperature. Trends in Plant Science 8,
343–351.
Banerjee, S., and Basu, P.S. 1992. Hormonal regulation of flowering and fruit
development: Effect of gibberellic acid and ethrel on fruit setting and
development of Momordica charantia L. Biologia Plantarum 34, 63–70.
Ben-Yehoshua, S., and Rodov, V. 2003. Transpiration and water stress. In
Bartz, J.A., and Brecht, J.K. (eds.), Postharvest Physiology and Pathology
of Vegetables, 2nd ed. Marcel Dekker, New York, pp. 111–159.
Biale, J.B., Young, R.E., and Olmstead, A.J. 1954. Fruit respiration and ethyl-
ene production. Plant Physiology 29, 168–174.
Blankenship, S.M., and Dole, J.M. 2003. 1-Methylcyclopropene: A review.
Postharvest Biology and Technology 28, 1–25.
Bregoli, A.M., Fabbroni, C., Raimondi, V., and Costa, G. 2010. Improving
colour and size of apricot fruit by means of exogenous auxin applica-
tion. Acta Horticulturae 862, 365–372.
Bron, I.U., Ribeiro, R.V., Cavalini, F.C., Jacomino, A.P., and Trevisan, M.J.
2005. Temperature-related changes in respiration and Q10 coefficient of
guava. Scientia Agricola 62, 458–463.

72
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Brummell, D.A., Harpster, M.H., and Dunsmuir, P. 1999. Differential

­expression of expansion gene family members during growth and ripen-
ing of tomato fruit. Plant Molecular Biology 39, 161–191.
Buran, T.J., Sandhu, A.K., Azeredo, A.M., Bent, A.H., Williamson, J.G., and
Gu, L. 2012. Effects of exogenous abscisic acid on fruit quality, anti-
oxidant capacities, and phytochemical contents of southern high bush
blueberries. Food Chemistry 132, 1375–1381.
Burdon, J.N., Dori, S., Lomaniec, E., Marinansky, R., and Pesis, E. 1994.
The postharvest ripening of water stressed banana fruits. Journal of
Horticulture Science 69, 799–804.
Buron-Moles, G., Torres, R., Amoako-Andoh, F., Viñas, I., Teixidó, N., Usall,
J., Keulemans, W., and Davey, M.W. 2014. Analysis of changes in pro-
tein abundance after wounding in ‘Golden Delicious’ apples. Postharvest
Biology and Technology 87, 51–60.
Canli, F.A., and Orhan, H. 2009. Effects of preharvest gibberellic acid applica-
tions on fruit quality of ‘0900 Ziraat’ sweet cherry. HortTechnology 19,
127–129.
Cantín, C.M., Fidelibus, M.W., and Crisosto, C.H. 2007. Application of abscisic
acid (ABA) at veraison advanced red color development and maintained
postharvest quality of ‘Crimson Seedless’ grapes. Postharvest Biology
and Technology 46, 237–241.
Cantwell, M. 2002. Summary table of optimal handling conditions for fresh
produce. In Kader, A.A. (ed.), Postharvest Technology of Horticultural
Crops. Agriculture and Natural Resources Publication 3311. University
of California, Oakland, pp. 511–518.
Choi, C., Wiersma, P.A., Toivonen, P., and Kappel, F. 2002. Fruit growth,
firmness and cell wall hydrolytic enzyme activity during development
of sweet cherry fruit treated with gibberellic acid (GA3). Journal of
Horticultural Science and Biotechnology 77, 615–621.
Daminato, M., Guzzo, F., and Casadoro, G. 2013. A SHATTERPROOF-like
gene controls ripening in non-climacteric strawberries, and auxin
and abscisic acid antagonistically affect its expression. Journal of
Experimental Botany 64, 3775–3786.
Daniels-Lake, B.J., Prange, R.K., Nowak, J., Asiedu, S.K., and Walsh, J.R. 2005.
Sprout development and processing quality changes in potato tubers
stored under ethylene. 1. Effects of ethylene concentration. American
Journal of Potato Research 82, 389–397.
Devoghalaere, F., Doucen, T., Guitton, B., Keeling, J., Payne, W., Ling, T.J.,
Ross, J.J., Hallett, I.C., Gunaseelan, K., Dayatilake, G.A., Diak, R., Breen,
K.C., Tustin, D.S., Costes, E., Chagné, D., Schaffer, R.J., and David,
K.M. 2012. A genomics approach to understanding the role of auxin in
apple (Malus × domestica) fruit size control. BMC Plant Biology 12, 7.
Dhanapal, A.P., Martínez-García, P.J., Gradziel, T.M., and Crisosto, C.H.
2012. First genetic linkage map of chilling injury susceptibility in peach
(Prunus persica (L.) Batsch) fruit with SSR and SNP markers. Journal of
Plant Science and Molecular Breeding, doi: 10.7243/2050-2389-1-3.

73
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Ding, J., Chen, B., Xia, X., Mao, W., Shi, K., Zhou, Y., and Yu, J. 2013. Cytokinin-
induced parthenocarpic fruit development in tomato in partly depen-
dent on enhanced gibberellin and auxin biosynthesis. PLoS ONE  8,
e70080. doi: 10.1371/journal.pone.0070080.
Fidelibus, M.W., Koch, K.E., and Davies, F.S. 2008. Gibberellic acid alters
sucrose, hexoses, and their gradients in peel tissues during color
break delay in ‘Hamlin’ orange. Journal of the American Society for
Horticultural Science 133, 760–767.
Gross, K.C., Wang, C.Y., and Saltveit, M. 2004. The Commercial Storage of
Fruits, Vegetables, and Florist and Nursery Stocks. USDA Handbook 66.
U.S. Department of Agriculture, Agricultural Research Service,
Beltsville, MD. http://www.ba.ars.usda.gov/hb66/ (accessed October 1,
2013).
Hernandez, O., Torres, C.A., Moya-León, M.A., Opazo, M.C., and Razmilic, I.
2014. Roles of the ascorbate-gluathione cycle, pigments and phenolics
in postharvest ‘sunscald’ development on ‘Granny Smith’ apples (Malus
domestica Borkh.). Postharvest Biology and Technology 87, 79–87.
Hiwasa, K., Rose, J.K.C., Nakano, R., Inaba, A., and Kubo, Y. 2003. Differential
expression of seven α-expansin genes during growth and ripening of
pear fruit. Physiologia Plantarum 117, 564–572.
Holcroft, D.M., and Kader, A.A. 1999. Controlled atmosphere-induced
changes in pH and organic acid metabolism may affect color of stored
strawberry fruit. Postharvest Biology and Technology 17, 19–32.
Hurkman, W.J., and Kennedy, G.S. 1975. Ultrastructural changes of chlo-
roplasts in aging tobacco leaves. Proceedings of Indiana Academy of
Sciences 85, 89–95.
Iglesias, D.J., Cercós, M., Colmenero-Flores, J.M., Naranjo, M.A., Ríos, G.,
Carrera, E., Ruiz-Rivero, O., Lliso, I., Morillon, R., Tadeo, F.R., and
Talon, M. 2007. Physiology of citrus fruiting. Brazilian Journal of Plant
Physiology 19, 333–362.
Inari, T., Yamauhi, R., Kato, K., and Takeuchi, T. 2002. Changes in pectic poly-
saccharides during the ripening of cherry tomato fruits. Food Science
and Technology Research 8, 55–58.
Jan, I., and Rab, A. 2012. Influence of storage duration on physic-­chemical
changes in fruit of apple cultivars. Journal of Animal and Plant
Science 22, 708–714.
Jiang, Y., and Joyce, D.C. 2003. ABA effects on ethylene production, PAL
activity, anthocyanin and phenolic contents of strawberry fruit. Plant
Growth Regulation 39, 171–174.
Kader, A.A., and Saltveit, M.E. 2003a. Atmosphere modification. In Bartz,
J.A., and Brecht, J.K. (eds.), Postharvest Physiology and Pathology of
Vegetables. Marcel Dekker, New York, pp. 229–246.
Kader, A.A., and Saltveit, M.E. 2003b. Respiration and gas exchange. In
Bartz, J.A., and Brecht, J.K. (eds.), Postharvest Physiology and Pathology
of Vegetables. Marcel Dekker, New York, pp. 7–29.

74
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Klein, J.D., and Goldshmidt, E.E. 2005. Hormonal regulation of ripening

and senescence phenomena. In Ben-Yehoshua (ed.), Environmentally
Friendly Technologies for Agricultural Produce Quality. Taylor & Francis
Group, Boca Raton, FL, pp. 315–331.
Kou, L., Yang, T., Luo, Y., Liu, X., Huang, L., and Codling, E. 2014. Pre-harvest
calcium application increases biomass and delays senescence of broc-
coli microgreens. Postharvest Biology and Technology 87, 70–78.
Kubo, Y., Xue, Y.B., Nakatsuka, A., Mathooko, F.M., Inaba, A., and Nakamura,
R. 2000. Expression of a water stress-induced polygalacturonase
gene in harvested cucumber fruit. Journal of the Japanese Society for
Horticultural Science 69, 273–279.
Kuckenberg, J., Tarachnyk, I., and Noga, G. 2008. Evaluation of fluorescence
and remission techniques for monitoring changes in peel chlorophyll
and internal fruit characteristics in sunlight and shaded sides of apple
fruit during shelf life. Postharvest Biology and Technology 48, 231–241.
Kulaeva, O.N., Karavaiko, N.N., Selivankina, S.Y., Kusnetsov, V.V.,
Zemlyachenko, Y.V., Cherepneva, G.N., Maslova, G.G., Lukevich, T.V.,
Smith, A.R., and Hall, M.A. 2000. Nuclear and chloroplast cytokinin-
binding proteins from barley leaves participating in transcription regu-
lation. Plant Growth Regulation 32, 329–335.
Lammertyn, J., Franck, C., Verlinden, B.E., and Nicolaï, B.M. 2001.
Comparative study of O2, CO2 and temperature effect on respiration
between ‘Conference’ pear cell protoplasts in suspension and intact
pears. Journal of Experimental Botany 52, 1769–1777.
Ludford, P.M. 2003. Hormonal changes during postharvest. In Bartz, J.A., and
Brecht, J.K. (eds.), Postharvest Physiology and Pathology of Vegetables.
Marcel Dekker, New York, pp. 31–77.
Luengwilai, K., Beckles, D.M., and Saltveit, M.E. 2012. Chilling-injury of har-
vested tomato (Solanum lycopersicum L.) cv. Micro-Tom fruit is reduced
by temperature pre-treatments. Postharvest Biology and Technology 63,
123–128.
Lurie, S., Shapiro, B., and Ben-Yehoshua, S. 1986. Effects of water stress and
degree of ripeness on rate of senescence of harvested bell pepper fruit.
Journal of the American Society for Horticultural Science 111, 880–885.
Martínez-García, P.J., Peace, C.P., Parfitt, D.E., Ogundiwin, E.A., Fresnedo-
Ramírez, J., Dandekar, A.M., Gradziel, T.M., and Crisosto, C.H. 2012.
Influence of year and genetic factors on chilling injury susceptibility in
peach (Prunus persica (L.) Batsch). Euphytica 185, 267–280.
Matsuo, S., Kikuchi, K., Fukuda, M., Honda, I., and Imanishi, S. 2012. Roles
and regulation of cytokinins in tomato fruit development. Journal of
Experimental Botany 63, 5569–5579.
Mattoo, A.K., and Lieberman, M. 1977. Localization of the ethylene-­
synthesizing system in apple tissue. Plant Physiology 60, 794–799.
Morris, L.L. 1982. Chilling injury of horticultural crops: An overview.
HortScience 17, 161–162.

75
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Nishiyama, K., Guis, M., Rose, J.K.C., Kubo, Y., Bennett, K.A., Wangjin, L.,
Kato, K., Ushijima, K., Nakano, R., Inaba, A., Bouzayen, M., Latché, A.,
Pech, J.-C., and Bennett, A.B. 2007. Ethylene regulation of fruit softening
and cell wall disassembly in Charentais melon. Journal of Experimental
Botany 58, 1281–1290.
Nunes, M.C.N., and Emond, J.P. 2003. Storage temperature. In Bartz, J.A., and
Brecht, J.K. (eds.), Postharvest Physiology and Pathology of Vegetables.
Marcel Dekker, New York, pp. 209–228.
Palmer, J.W., Harker, F.R., Tustin, D.S., and Johnston, J. 2010. Fruit dry ­matter
concentration: A new quality metric for apples. Journal of the Science of
Food and Agriculture 90, 2586–2594.
Pech, J.C., Bouzayen, M., and Latché, A. 2008. Climacteric fruit ripening:
Ethylene-dependent and independent regulation of ripening pathways
in melon fruit. Plant Science 175, 114–120.
Pedreschi, R., Hertog, M., Robben, J., Lilley, K.S., Karp, N.A., Baggerman, G.,
Vanderleyden, J., and Nicolaï, B. 2009. Gel-based proteomics approach
to the study of metabolic changes in pear tissue during storage. Journal
of Agricultural and Food Chemistry 57, 6997–7004.
Pilkington, S.M., Montefiori, M., Galer, A.L., Emery, R.J.M., Allan, A.C., and
Jameson, P.E. 2013. Endogenous cyctokinin in developing kiwifruit is
implicated in maintaining fruit flesh chlorophyll levels. Annals of Botany
112, 57–68.
Pollock, C.J., and ap Rees, T. 1975a. Activities of enzymes of sugar metabolism
in cold-stored tubers of Solanum tuberosum. Phytochemistry 14, 613–617.
Pollock, C.J., and ap Rees, T. 1975b. Effect of cold on glucose metabolism by
callus and tubers of Solanum tuberosum. Phytochemistry 14, 1903–1906.
Qian, C., He, Z., Zhao, Y., Mi, H., Chen, X., and Mao, L. 2013. Maturity-
dependent chilling tolerance regulated by the antioxidative capacity in
postharvest cucumber (Cucumis sativus L.) fruits. Journal of the Science
of Food and Agriculture 93, 626–633.
Reid, M.S. 2002. Ethylene in postharvest technology. In Kader, A.A. (ed.),
Postharvest Technology of Horticultural Crops. Agriculture and Natural
Resources Publication 3311. University of California, Oakland,
pp. 149–162.
Rhodes, M.J.C., Wooltorton, L.S.C., and Hill, A.C. 1981. Changes in phenolic
metabolism in fruit and vegetable tissues under stress. In Friend, J., and
Rhodes, M.J.C. (eds.), Recent Advances in the Biochemistry of Fruits and
Vegetables. Academic Press, London, pp. 193–220.
Rudell, D.R., Buchanan, D.A., Leisso, R.S., Whitaker, B.D., Mattheis, J.P.,
Zhu, Y., and Varanasi, V. 2011. Ripening, storage temperature, ethylene
action, and oxidative stress alter apple peel phytosterol metabolism.
Phytochemistry 72, 1328–1340.
Rudell, D.R., Mattheis, J.P., and Hertog, M.L. 2009. Metabolomic change
precedes apple superficial scald symptoms. Journal of Agricultural and
Food Chemistry 57, 8459–8466.

76
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Saltveit, M.E. 1999. Effect of ethylene on quality of fresh fruits and vegeta-
bles. Postharvest Biology and Technology 15, 279–292.
Saltveit, M.E. 2003. Is it possible to find an optimal controlled atmosphere?
Postharvest Biology and Technology 27, 3–13.
Saltveit, M.E. 2004. Respiratory metabolism. In Gross, K.C., Wang, C.Y.,
and Saltveit, M.E. (eds.), The Commercial Storage of Fruits, Vegetables,
and Florist and Nursery. USDA Handbook 66. U.S. Department of
Agriculture, Agricultural Research Service, Beltsville, MD. http://www.
ba.ars.usda.gov/hb66/019respiration.pdf (accessed October 15, 2013).
Sandhu, A.K., Gray, D.J., Lu, J., and Gu, L. 2011. Effects of exogenous
abscisic acid on antioxidant capacities, anthocyanins, and flavonol con-
tents of muscadine grape (Vitis rotundifolia) skins. Food Chemistry 126,
982–988.
Saquet, A.A., Streif, J., and Bangerth, F. 2000. Changes in ATP, ADP and
pyridine nucleotide levels related to the incidence of physiological dis-
orders in ‘Conference’ pears and ‘Jonagold’ apples during controlled
atmosphere storage. Journal of Horticultural Science and Biotechnology
75, 243–249.
Sargent, S.A., and Moretti, C.L. 2004. Tomato. In Gross, K.C., Wang, C.Y.,
and Saltveit, M.E. (eds.), The Commercial Storage of Fruits, Vegetables,
and Florist and Nursery. USDA Handbook 66. U.S. Department of
Agriculture, Agricultural Research Service, Beltsville, MD. http://www.
ba.ars.usda.gov/hb66/138tomato.pdf (accessed October 15, 2013).
Sevillano, L., Sanchez-Ballesta, M.T., Romojaro, F., and Flores, F.B. 2009.
Physiological, hormonal and molecular mechanisms regulating chill-
ing injury in horticultural species. Postharvest technologies applied
to result its impact. Journal of the Science of Food and Agriculture 89,
555–573.
Singh, K., Weaver, R.J., and Johnson, J.O. 1978. Effect of applications of
gibberellic acid on berry size, shatter, and texture of Thompson
­
­seedless  grapes. American Journal of Enology and Viticulture 29,
258–262.
Sugar, D., and Basile, S.R. 2013. Integrated ethylene and temperature condi-
tioning for induction of ripening capacity in ‘Anjou’ and ‘Comice’ pears.
Postharvest Biology and Technology 83, 9–16.
Sun, Y., Chen, P., Duan, C., Tao, P., Wang, Y., Ji, K., Hu, Y., Li, Q., Dai, S.,
Wu, Y., Luo, H., Sun, L., and Leng, P. 2013. Transcriptional regulation of
genes encoding key enzymes of abscisic acid metabolism during melon
(Cucumis melo L.) fruit development and ripening. Journal of Plant
Growth Regulation 32, 233–244.
Tacken, E., Ireland, H., Gunaseelan, K., Karunairetnam, S., Wang, D., Shultz,
O., Bowen, J., Atkinson, R.G., Johnston, J.W., Putterill, J., Hellens, R.P.,
and Schaffer, R.J. 2010. The role of ethylene and cold temperature in
the regulation of the apple POLYGALACTURONASE1 gene and fruit
softening. Plant Physiology 153, 294–305.

77
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Thompson, J., Kader, A., and Sylva, K. 1996. Compatibility chart for fruits and
vegetables in short-term transport or storage. Agriculture and Natural
Resources Publication 21560. University of California, Oakland.
Timbie, M., and Haard, N.F. 1977. Involvement of ethylene in the hardcore
syndrome of sweet potato roots. Journal of Food Science 42, 491–493.
Toivonen, P.M.A. 2003. Effects of storage conditions and postharvest pro-
cedures on oxidative stress in fruits and vegetables. In Hodges, D.M.
(ed.), Postharvest Oxidative Stress in Horticultural Crops. Food Products
Press, New York, pp. 69–90.
Toivonen, P.M.A. 2004. Postharvest storage procedures and oxidative stress.
HortScience 39, 938–942.
Toivonen, P.M.A. 2010. Postharvest physiology of vegetables. In Hui, Y.H.,
Sinha, N., Ahmed, J., Evranuz, E.Ö., and Siddiq, M. (eds.), Handbook of
Vegetables and Vegetable Processing. Wiley-Blackwell Publishing, Ames,
IA, pp. 199–220.
Toivonen, P.M.A., and DeEll, J.R. 2002. Physiology of fresh-cut fruits and veg-
etables. In Lamikanra, O. (ed.), Fresh-Cut Fruits and Vegetables: Science,
Technology, and Market. CRC Press, Boca Raton, FL, pp. 91–123.
Toivonen, P.M.A., and Hodges, D.M. 2011. Abiotic stress in harvested fruits
and vegetables. In Venkteswarlu, B., and Shanker, A.K. (eds.), Abiotic
Stress in Plants—Mechanisms and Adaptations. Intech Open Access
Publishers, Rijeka, Croatia, pp. 39–59.
Toivonen, P.M.A., Hampson, C.R., Stan, S., McKenzie, D.-L., and Hocking,
R. 2007. Factors affecting severity of bruises and degree of apparent
bruise recovery in a yellow-skinned apple. Postharvest Biology and
Technology 45, 276–280.
Vinocur, B., and Altman, A. 2005. Recent advances in engineering plant toler-
ance to abiotic stress: Achievements and limitations. Current Opinion in
Biotechnology 16, 123–132.
Wang, C.Y. 1982. Physiological and biochemical responses of plants to chill-
ing stress. HortScience 17, 173–186.
Wang, C.Y. 1993. Approaches to reduce chilling injury of fruits and vegeta-
bles. Horticultural Reviews 15, 63–95.
Wang, C.Y. 2004. Chilling and freezing injury. In Gross, K.C., Wang, C.Y.,
and Saltveit, M.E. (eds.), The Commercial Storage of Fruits, Vegetables,
and Florist and Nursery. USDA Handbook 66. U.S. Department of
Agriculture, Agricultural Research Service, Beltsville, MD. http://www.
ba.ars.usda.gov/hb66/018chilling.pdf (accessed October 15, 2013).
Wang, C.Y., and Qi, L. 1997. Modified atmosphere packaging alleviates chill-
ing injury in cucumbers. Postharvest Biology and Technology 10, 195–200.
Wang, H., Qian, S., Ma, S., Zhou, Y., Patrick, J.W., Duan, X., Jiang, Y., and Qu,
H. 2013. Energy status of ripening and postharvest senescent fruit of
litchi (Litchi chinensis Sonn.). BMC Plant Biology 13, 55.
Wang, K.L.-C., Li, H., and Ecker, J.R. 2002. Ethylene biosynthesis and signal-
ing networks. The Plant Cell (Suppl.), S131–S151.

78
P o s t h a r v e s t P hy s i o l o g y o f F r u i t s a n d V e g e t a b l e s

Watada, A.E., Herner, R.C., Kader, A.A., Romani, R.J., and Staby, G.L. 1984.
Terminology for the description of developmental stages of horticul-
tural crops. HortScience 119, 20–21.
Watkins, C.B. 2008. Dynamic controlled atmosphere storage: A new technol-
ogy for the New York storage industry? New York Fruit Quarterly 16,
23–26.
Watkins, C.B., Kupferman, E., and Rosenberger, D.A. 2004. Apples. In
Gross, K.C., Wang, C.Y., and Saltveit, M.E. (eds.), The Commercial
Storage of  Fruits, Vegetables, and Florist and Nursery. USDA Handbook
66. U.S.  Department  of Agriculture, Agricultural Research Service,
Beltsville, MD. http://www.ba.ars.usda.gov/hb66/027apple.pdf (accessed
October 15, 2013).
Whitaker, B.D. 2004. Oxidative stress and superficial scald of apple fruit.
HortScience 39, 933–937.
Wills, R.B.H., Ku, V.V.V., Shohet, D., and Kim, G.H. 1999. Importance of low
ethylene levels to delay senescence of non-climacteric fruit and vegeta-
bles. Australian Journal of Experimental Agriculture 39, 221–224.
Wills, R.B.H., Warton, M.A., and Ku, V.V.V. 2000. Ethylene levels associ-
ated with fruit and vegetables during marketing. Australian Journal of
Experimental Agriculture 40, 465–470.
Zhang, C., Tanabe, K., Tani, H., Nakajima, H., Mori, M., and Sakuno, E. 2007.
Biologically active gibberellins and abscisic acid in fruit of two later-
maturing Japanese pear cultivars with contrasting fruit size. Journal of
the American Society for Horticultural Science 132, 452–458.
Zhang, Z., Fu, R., Huber, D.J., Rao, J., Chang, X., Hu, M., Zhang, Y., and Jiang,
N. 2012. Expression of expansin gene (CDK-Exp3) and its modulation
by exogenous gibberellic acid during ripening and softening of persim-
mon fruit. HortScience 47, 378–381.
Zubo, Y.O., Yamburenko, M.V., Selivankina, S.Y., Shakirova, F.M., Avalbaev,
A.M., Kudryakova, N.V., Zubkova, N.K., Liere, K., Kulaeva, O.N.,
Kusnetsov, V.V., and Börner, T. 2008. Cytokinin stimulates chloroplast
transcription in detached barley leaves. Plant Physiology 148, 1082–1093.
Zuo, J., Zhu, B., Fu, D., Zhu, Y., Ma, Y., Chi, L., Ju, Z., Wang, Y., Zhai, B., and
Luo, Y. 2012. Sculpting the maturation, softening and ethylene pathway:
The influences of microRNAs on tomato fruits. BMC Genomics 13, 7.

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Chapter 3

Postharvest Quality
of Ornamental Plants
Fernando L. Finger, Tania P. Silva, Fernanda
F. Araujo, and Jose G. Barbosa
Federal University of Viçosa, Viçosa, Minas Gerais, Brazil

Abstract 82
3.1 Introduction 82
3.2 Quality Attributes in Ornamental Plants 83
3.3 Influence of Water Relations on Ornamental Longevity 85
3.4 Action of Ethylene on Ornamental Plant Quality 88
3.4.1 Inhibition of Ethylene Synthesis 91
3.4.2 Inhibition of Ethylene Action 93
3.4.3 Ethylene Absorbers 95
3.5 Role of Abscisic Acid, Gibberellins, and Cytokinins 96
3.6 Role of Calcium on Flower Senescence 97
3.7 Respiration 98
3.8 Temperature 99
3.9 Handling of Cut Flowers 102
3.10 Potted Plants 103
3.11 Conclusions and Future Perspectives 104
Acknowledgment 105
References 105

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Abstract
In this chapter, the behavior of cut flowers and potted ornamental plants is
studied regarding the physiological and environmental factors that affect
the rate of senescence and longevity. Water uptake by cut flowers, carbohy-
drate supply, and response of flowers to ethylene interact alone or together
to affect the length of the vase life. Depending on whether ethylene is the
main cause of flower senescence, particular handling is required to extend
vase life. Periodically, recuts of the flower stem or pulsing with sucrose
improves the water uptake and floret opening of the bird-of-paradise. Any
flower species or potted plants with high sensitivity to ethylene have a
much better shelf life when treated with inhibitors of ethylene action such
as 1-methylcyclopropene (1-MCP) or silver thiosulfate (STS). The orchid
Epidendrum ibaguense had a positive response by reducing flower abscis-
sion when treated with the aminoethoxyvivylglycine (AVG) inhibitor of
­ethylene synthesis. But in those flowers insensitive to ethylene presence,
water status and carbohydrate supply play a very important role in the
length of the vase life. Thus, pulsing solutions containing sugar or STS
may be effective in postponing earlier senescence in many flower species.
Based on the rate of leaf yellowing and abscission of fruits and leaves, treat-
ment of potted ornamental peppers with 1-MCP prolongs postproduction
life in indoor conditions. However, ethylene partially reversed the inhibition
of 1-MCP on leaf yellowing and abscission.

3.1 Introduction
Ornamental plants, in general, have limited vase or postproduction life;
therefore, the use of adequate postharvest practices of handling is essential
to maintain the quality and prolong the usefulness of the potted plant or
flower. The applied techniques should allow for transportation, and for the
majority of ornamental species, relatively short-term storage—at wholesale
or retail stores—is required before reaching the final consumer.
From the consumer’s point of view, quality is associated with the length
of the shelf life, either at home or in display areas. But the shelf life varies
from ornamental plant to plant, as well as the initial quality of the product
and previous treatments applied to extend the postharvest life. Particularly
in underdeveloped and developing countries, the production areas of orna-
mental plants are not far from the selling commercial centers, and even with-
out modern postharvest handling, the final consumer still receives products
of reasonable quality. However, with the rapid increase in urban population,
the production areas have moved farther away. Under such a scenario, the
use of new techniques for preservation is required, which must guarantee

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Postharvest Quality of Ornamental Pl ants

quality to the florist and be satisfactory to the ­consumer. Among the s­everal
techniques available for ornamental plant storage, temperature, relative
humidity, and composition of the atmosphere must be addressed. In addi-
tion, florists have to incorporate treatments to reduce the water and tem-
perature stresses, avoid the deleterious effects of ethylene, and minimize
the influence of low-level radiation in the storage and display areas.
After harvesting, the cut flowers or the potted plants at the postpro-
duction phase are subjected to a series of abiotic stresses, including water
stresses, intense transpiration, exposure to ethylene, and development of
physiological disorders. Usually, leafy ornamental plants are more resis-
tant to senescence than flowers. Flowers, on the other hand, due to their
ephemeral nature and the reduced supply of organic substances, find that
the catabolic metabolism accelerates quickly, thus altering important phys-
iological processes, such as reduction of the water uptake rate, depletion
of respiratory substrates, and an increase of ethylene production and sen-
sitivity. Despite the important influence of ethylene in reducing the vase
life of many ornamental crops, ornamental plants may respond to other
hormones, such as abscisic acid, gibberellins, and cytokinins, to extend of
their postharvest life.
Like any fresh product, ornamental plants, once harvested or trans-
ferred to the display areas, maintain intense respiratory and transpiratory
activities. Respiration will deplete the reserves of organic substrates, which
are already limited, whereas intense transpiration will result in the fast wilt-
ing of leaves and flowers.
Production of carbon dioxide by the ornamental plants can be dimin-
ished by reducing the storage temperature, usually as low as possible, but
above the freezing point of the cells. Many ornamental plants, however,
originate from tropical and subtropical climates and are susceptible to
chilling injury, which restricts the length of shipping and storage at low
temperatures.
In this chapter, we address the different aspects involved in the pres-
ervation of potted ornamental plants and cut flowers, with special attention
to postharvest treatments available to diminish the rate of leaf and flower
senescence.

3.2  Quality Attributes in Ornamental Plants

For most ornamental plants, there is no government-required quality
­g rading system. However, most florist associations voluntarily establish
grading standards, mainly for cut flowers and foliage. As a general rule, for
cut flowers, emphasis is given to the uniformity of the plant material, focus-
ing on the stem length and diameter and the presence of curved portions.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Regarding the flower itself, the most important attributes are the size of the
bud, stage of development, and lack of wilted or abscised petals.
Two important criteria must be achieved at harvest for any ornamen-
tal plant: a presence quality compatible with the consumer’s wishes and an
extended shelf life after harvest or postproduction of potted plants, which
will allow enough time for transportation, eventual storage, and final dis-
play. In order to reach these goals, ornamental plants have to be harvested
at a specific stage of development, as presented in Table 3.1.
The establishment of the developmental stage for harvesting a
­particular flower is based on the ability of its bud flower to open after har-
vest, its response to abiotic stresses, and the length of its shelf life.
The standard quality for potted plants depends on the plant s­ pecies.
For instance, for chrysanthemum and ornamental peppers, the plant canopy

Table 3.1  Stage of Flower Development at Harvest

Scientific Name Common Name Development at Harvest
Anthurium cultorum Anthurium Spadix 3/4 fully developed
Antirrhinum majus Snapdragon Inflorescence with 1/3 to 2/3 of
open florets
Cattleya hybrids Orchid Fully open flower
Delphinium ajacis Delphinium Inflorescence with 1/2 of open
florets
Dendrobium spp. Dendrobium Almost fully open flowers
Dianthus caryophyllus Carnation From half to fully open flower
Eustoma grandiflorum Lisianthus Oldest flower from the stalk fully
open
Gerbera hybrids Gerbera When two outer disc rows
are open
Gladiolus hybrids Gladiolus When few florets are showing
color
Gypsophila Baby’s breath Flowers fully open
paniculata
Heliconia spp. Heliconia Harvest at stage of final display
Iris hybrids Dutch iris Flower buds showing color
Lilium spp. Asiatic lily Lower buds showing color
Rosa hybrids Rose Beginning of the bud petal
to unfold
Strelitzia reginae Bird-of-paradise From first floret showing color
to fully open
Zinnia elegans Zinnia Almost fully open flower

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Postharvest Quality of Ornamental Pl ants

should cover the whole surface area of the pot. Also, shorter plants with
a compact canopy have a better appearance than taller ones. But in pots
containing orchids and African violets, the presence of flowers in different
stages of opening and the absence of senescent flowers or leaves are more
important than the shape of the plant canopy.

3.3  Influence of Water Relations

on Ornamental Longevity
Harvested ornamentals plants, in particular cut flowers, usually present an
imbalance between water uptake and transpiration throughout the vase life,
which results in the wilting of petals and premature senescence. Several
causes can be attributed to the reduction of water absorption by the flow-
ers, including obstruction of the xylem by microorganisms, deposition of
pectin and phenols, and air embolism (Van Doorn, 1997). Cut carnations
and roses present a reduction of longevity due to an imbalance of water
status, which is related to the diminished capacity of water uptake by the
cut flower ­during the vase life. The reduction of the water uptake rate
accounts for the continuous drop of the fresh weight of the flower during
the vase life. Van Doorn et al. (1995) found the presence of Pseudomonas
spp., Acinetobacter calcoaceticus, and Alcaligenes spp. bacteria in a vase of
carnations after 10 days at room temperature. The presence of bacteria in
the water caused obstruction of water uptake by the stem and reduced the
longevity, which was inversely proportional to an increased bacterial count
in the vase water. However, regardless of the cause of vase blockage, the
hydraulic conductance is diminished throughout the vase life of the flower,
establishing disequilibrium between the water uptake and transpiration. In
Zinia elegans, Carneiro et al. (2002) observed that one of the first symptoms
of reduced water uptake by the stem was a decrease of its fresh weight,
which began within the first 24 h of harvest. However, when the base of
the stem was recut at a frequency of every 12 h, the weight loss rate was
diminished compared to that of the uncut flower control. This simple action
of recutting the base of the stem improved the flower water balance, keep-
ing the whole flower with a higher water content (Figure 3.1). But in flowers
like Eustoma grandiflorum, placed in similar environmental conditions, a
noticeable decrease was seen in the fresh weight of the whole stem over a
much longer period of time, after only 6 days in the vase (Liao et al., 2001).
Due to the rapid weight loss, Z. elegans flowers rapidly show symptoms of
wilting, in both petals and leaves; a delay of weight loss with concomitant
extension of its longevity was achieved by recutting the base of the stem. In
E. ­grandiflorum and cut roses, an extension of the longevity can be obtained
by adding 250 mg L −1 aluminum sulfate, a well-known antimicrobial agent.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

105

100
Fresh weight (%)

95

90

85

80
0 20 40 60 80 100 120
Hours after harvest

Figure 3.1  Behavior of fresh weight in Zinnia elegans cut flowers recut ( )
and uncut ( ) at base of the stem every 12 hours. (From Carneiro, T.F. et al.,
Pesquisa Agropecuaria Brasileira 37, 1065–1070, 2002.)

Also, the inclusion of 200 mg L −1 8-hydroxyquinoline citrate or sulfate in

the vase solution extends the vase life of roses, due to less bacterial growth
in the water.
Once the bird-of-paradise is harvested, the content of water in the
flower keeps diminishing during the vase life, due to physical blockage at
the base of the stem. Apparently, the blockage of the xylem is due to the
high activity of peroxidase, a group of isozymes responsible for deposit-
ing complexes of oxidized phenolic compounds outside of the cell wall. In
this flower, the water status in the tissues can be maintained unchanged
by recutting the stem base by 2 cm every 2 days (Figure 3.2). Such a pro-
cedure keeps the relative water content of the sepal close to 93.9% during
the vase life of the flower, while in uncut stems the water content drops to
approximately 86% (Figure 3.2). As a result, the cutting of the stem base
extends the vase life of the flower by at least 1 day compared to that of
uncut ones. In other cut flowers such as chrysanthemum, xylem occlusion
in the base of the stem occurs due to the joint activity of peroxidase and
polyphenoloxidase; both enzymes are involved in the biosynthesis of lignin

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Postharvest Quality of Ornamental Pl ants

96

94
Relative water contet (%)

92

90

88

86
80
0 2 4 6 8 10
Days

Figure 3.2  Relative water content in the sepals of bird-of-paradise flower

recut every 2 days ( ) and uncut ( ) maintained at 25°C and 60% rela-
tive humidity. (From Campanha, M.M. et al., Revista Brasileira Horticultura
Ornamental 3, 27–31, 1997.)

and suberin (Van Doorn and Vaslier, 2002). But when inhibitors for either
enzyme were applied to vase water, a delayed blockage of the xylem vessels
was observed, resulting in a longer time until leaf wilting for the cut chry-
santhemum flower.
In roses, the vascular occlusion in the xylem is primarily associated
with the presence of bacteria in the cut at the base of the stem and by the
growing of bacteria in the water of the vase, blocking the water flow to the
flower due to the presence of extracellular polysaccharides and other degra-
dation products that originated from the dead bacteria (Van Doorn, 1997). In
such condition, the roses will develop several symptoms of water deficiency,
including lack of petal opening, bending of the stem neck below the flower,
and leaf wilting (Bleeksma and Van Doorn, 2003). But when roses were main-
tained in the vase in a solution containing 200 mg L −1 8-­hydroxyquinoline
sulfate alone or mixed with 30 g L −1 sucrose, the hydraulic conductance of
the stem was kept close to the initial level at harvest, and as consequence,

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flower longevity was prolonged (Ichimura et al., 1999). Furthermore, the

addition of sucrose to the solution provided longer vase life by increasing the
carbohydrate content in the petals.
Hard water contains minerals that turn the water pH to alkaline,
which diminishes its movement through the plant tissues. Such a problem
can be solved by removing the minerals from the water with a deionizer or
by lowering the pH, making the water acidic. The pH of hard water should
be lowered to 3.5–4.0 by adding citric acid to the vase water.

3.4  Action of Ethylene on Ornamental Plant Quality

Cut flowers or potted plants are often exposed to several kinds of stresses
once they are harvested or moved from the field or greenhouses. Different
abiotic stresses may occur during shipping, storage in wholesale ware-
houses, retail stores, and in the final display areas, usually in indoor envi-
ronments. Exposing plants to ethylene may induce a variety of effects, from
inducing senescence and abscission of flowers and leaves to impacting the
rate of flower opening. Some of the effects of ethylene on ornamental plants
are shown in Table 3.2. The toxic symptoms of ethylene action vary consid-
erably according to the species, but always induce deleterious effects that
reduce the display life.

Table 3.2  Symptoms of Ethylene Action in Ornamental Cut and Potted Plants
Plant Symptoms
Alstroemeria Darkening, petal abscission
Asiatic lily Abscission of flower, inhibition of flower opening
Carnation Flower wilting
Chrysanthemum Accelerates flower, leaf senescence
Delphinium ajacis Flower abscission
Gypsophila Flower wilting
paniculata
Iris Accelerates flower senescence
Orchids Flower abscission, senescence, epinasty, reddish petal
color
Ornamental Leaf yellowing; abscission of fruits, leaves, and flowers
peppers
Poinsettia Leaf and flower abscission, leaf epinasty
Roses Accelerates flower senescence, induces bud and petal
abscission, interruption of bud flower opening
Snapdragon Floret abscission

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Postharvest Quality of Ornamental Pl ants

Ethylene is produced by any living plant tissue, but it is also a by-

product of modern industrial human activity. In most ornamental plants,
undesirable effects can be ethylene induced, including petal wilting and
abscission, epinasty, and chlorophyll loss in leaves. Woltering and Van
Doorn (1988) concluded that flower species could be classified in three
types as follows: wilting mediated by ethylene, wilting not mediated by eth-
ylene, and petal abscission mediated by ethylene.
Synthesis of ethylene in plants is initiated from the amino acid
­methionine as follows: L-methionine → S-adenosyl methionine (SAM) →
amino-1-cyclopropane-1-carboxylic acid (ACC) → ethylene. The formation
of ACC from SAM and the oxidation of ACC to ethylene are catalyzed by
ACC synthase (ACS) and ACC oxidase (ACO), respectively. These two
enzymes play a major role in the regulation of ethylene production, being
stimulated by abiotic and biotic stresses. According to the changes in eth-
ylene production, activity of ACS and ACO, and behavior of respiration
during the development of a flower, it can be identified as climacteric, non-
climacteric, and in some cases, with an intermediate behavior. Carnation,
orchids, and Delphinium flowers have a typically climacteric behavior, char-
acterized by the autocatalytic production of ethylene and a parallel increase
in CO2 production. In flowers like roses and Narcissus, a nonclimacteric
senescence occurs during the senescence. Contrary to climacteric flowers,
in a nonclimacteric flower, such as Grevillia cv. ‘Sylvia’, a poor correlation is
observed among ethylene production, respiration, and changes in the con-
tent of ACC during senescence (Setyadjit et al., 2004). Nevertheless, the
climacteric increase of ethylene production and respiration varies among
the different cultivars from the same flower species, as well as the response
and sensitivity to ethylene. During the flower senescence of carnation and
other climacteric flowers, the autocatalytic induction of ethylene produc-
tion observed in the petals is due to an increase in the expression of mRNAs
from the ACC synthase and ACC oxidase.
The senescence in the majority of climacteric flowers can be rapidly
induced by the pollination phenomenon, observed in carnation, orchids,
and Petunia. In these flowers, pollination increases ethylene production,
inducing petal wilting and abscission. Prevention of these deleterious ethyl-
ene-induced effects can be achieved by emasculation or by pretreating the
flowers with antiethylene action compounds. Pollination of the cut orchid
Phalaenopsis induced a rapid senescence, characterized by the wilting of
the petals (Porat et al., 1994b). These same authors found that in a control
not pollinated, the time for the incipient wilting of Phalaenopsis flowers at
room temperature was 19.2 days, similar to that of the emasculated flowers,
while in pollinated flowers, the time for the wilting dropped to 2.0 days.
The pollination of Phalaenopsis flowers induced senescence of the orchid
by increasing autocatalytic ethylene production, as well as enhancing the
sensitivity to ethylene.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Flower species have different degrees of sensitivity to ethylene,

v­ arying from insensitive, such as roses, bird-of-paradise, and Gladiolus, to
very sensitive, including carnation and orchids. Nevertheless, the degree of
sensitivity to ethylene in the same species varies from cultivar to cultivar,
as observed in roses.
As shown in Table 3.3, the number of important commercial cut
­flowers sensitive to the presence of ethylene is superior to the number of
flowers with low sensitivity or ethylene insensitivity.
Species of flowers considered to be highly sensitive to ethylene show
physiological responses in the presence of relatively low concentrations of the
hormone, usually between 0.1 and 1.0 µl L −1 air, when exposed for a period of
6–12 h. When a cut orchid Phalaenopsis is exposed to ethylene at a concentra-
tion of 0.1 µl L −1 air for 12 h, the longevity of the flower is reduced by 4.7%,
and it increases to 27.9% for a concentration of 0.5 µl L −1 (Porat et al., 1994b).
The action of ethylene in the senescence of the orchid Epidendrum
ibaguense is affected by the increase in ethylene concentration. A fast
increase in flower abscission and reduction of longevity was present when
the cut flowers were sprayed with different concentrations of ethephon
(Figure  3.3). At a concentration of 0.1 mg L −1 ethephon, an exponential
increase in the rate of abscission is observed, reaching saturation for the
response at concentrations of 100 mg L −1 ethephon or above. As a conse-
quence, the longevity dropped from 6.8 days for the unsprayed flowers to
3 days for the 100 and 1000 mg L −1 ethephon treatments (Figure 3.3).

Table 3.3  Degree of Sensitivity to Ethylene in Commercial Flower Species

High Low Insensitive
Alstroemeria Anthurium Strelitzia reginae
Consolida ajacis (Dephinium) Aspargus Roses
Iris Gerbera Gladiolus
Gypsophila paniculata Tulipa hybrida Sandersonia
aurantiaca
Narcissus pseudonarcissus Waxflower Orchid (Cymbidium)
Orchids (Phalaenopsis, Chrysanthemum Buddleia davidii
Dendrobium, Epidendrum)
Petunia hybrida Cosmos bipinnatus
Carnation Cersis canadensis
Snapdragon Echinacea purpurea
Lily
Lathyrus odoratus
Portulaca
Hibiscus rosa-sinensis

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Postharvest Quality of Ornamental Pl ants

25 8

20
Abscission (flower day −1)

Longevity (days)
15

10

2
0 200 400 600 800 1000
Ethephon (mg l−1)

Figure 3.3  Influence of ethephon on abscission ( ) and longevity ( ) of

orchid Epidendrum ibaguense. (From Moraes, P.J. et al., Acta Horticulturae
813, 565–570, 2009.)

As previously determined for other orchid flowers, Epidendrum

­ibaguense can be classified as a high ethylene-sensitive species. But not
all orchid species are sensitive to ethylene, such as Cymbidium (Table 3.3),
which shows no increase in flower abscission when it is exposed to
­ethylene. Nonetheless, flower discoloration is hastened in the presence of
the ­hormone (Van Doorn, 2002).

3.4.1  Inhibition of Ethylene Synthesis

The activity of ACC synthase can be reduced by applying solutions con-
taining rhizobitoxin analogues, such as aminoethoxyvivylglycine (AVG)
or aminooxyacetic acid (AOA). However, neither inhibitor can completely
knock down the production of ethylene. Such inhibitors have been tested
in both preharvest sprays and postharvest vase solutions and sprays, but
the efficiency of these treatments varies considerably among the flowers.
In orchid Dendrobium, the addition of 0.5 mM AOA to the vase solution
delayed the time to complete petal wilting and blocked the ethylene pro-
duction induced by the flower pollination (Porat et al., 1994a). On the other
hand, in Gypsophyla paniculata, another high ethylene-sensitive flower,

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

like the orchid Dendrobium, the use of commercial preservatives contain-

ing low concentrations of AVG had no effect compared to a control treat-
ment with no antiethylene substances (Newman et al., 1998). Commercial
AVG registered by the name of Retain® has been used in several countries
to control the natural senescence of ornamentals and fresh fruits and veg-
etables. When Retain was tested as a pulsing solution for at least 6 hours
or by spraying the inflorescence until it ran off cut Epidendrum ibaguense
flowers, a significant reduction of flower abscission was observed, which
resulted in extended vase life compared to that of the control treatment
(Table 3.4). Regardless of the way in which the AVG was applied, con-
centrations of 1.5 and 2.0 mM were more efficient in reducing the overall
flower abscission. Nonetheless, the spray was much more effective than
treating the flowers by pulsing. This difference in efficiency between the
two methods might be related to the faster uptake by the flower when a
spray solution is used.
The vase life of cut flowers, especially those sensitive to ethylene,
is extended by treating them with substances that can block the ethylene
action or reduce the hormone production. But in most cases, the inhibitors
of ethylene action seem to be more efficient in prolonging the longevity
of the flowers than the substances responsible for reducing the activity of
the key enzymes from ethylene synthesis, ACC synthase and ACC oxidase.
These obtain better results by the use of ethylene action blockers, in part
because the inhibitors of ACC synthase or ACC oxidase do not protect the
flower from action of exogenous ethylene produced from sources other than
the plant’s own tissue. Furthermore, as the AVG and AOA do not completely
knock down the production of ethylene, the remaining small ethylene pro-
duced by the flower can accelerate the senescence in sensitive ornamentals.
Maintaining roses in vase solution containing 5% sucrose plus 0.5,
1.0, 1.5, and 2.0 mM AOA significantly improved the longevity compared

Table 3.4  Percentage of Flower Abscission in Epidendrum ibaguense Pulsed

or Sprayed with AVG
AVG (mM) Pulsinga Spray
0.5 69.7 Ba 45.8 Ab
1.0 84.5 Aa 20.5 Bb
1.5 56.1 Ca 18.6 Cb
2.0 54.0 Da 18.5 Cb
Source: Mapeli, A.M. et al., Pesquisa Agropecuaria Brasileira 44, 258–262,
2009.
a Pulsing for at least 6 hours. Mean separation within columns by Tukey’s test;

values followed by the same uppercase and lowercase letters are not signifi-
cantly ­different at the 5% level.

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Postharvest Quality of Ornamental Pl ants

to 5%  sucrose alone or control with distilled water, regardless of the

­concentration of AOA applied (Ketsa and Narkbua, 2001). However, the
authors did not measure the behavior of ethylene or CO2 production of
the  flowers in the presence of AOA. Based on their results, the addition
of AOA lowered the vase solution pH from 4.5 to 2.9, resulting in less growth
of bacteria in the solution, which might improve the water uptake and reduce
the development of bent neck.
Another inhibitor of ethylene production is acetylsalicylic acid (ASA),
which acts on the ACC oxidase enzyme. Fan et al. (1996) found that apple
ACC oxidase was strongly inhibited by ASA in both in vitro and in vivo
assays, as well as diminishing the fruit respiration. However, only a few
attempts have been made to evaluate the effectiveness of ASA in delay-
ing the deleterious effect of ethylene in cut flowers. When cut flowers of
Consolida ajacis were sprayed with 2 mM AOA or 20 mM ASA, they showed
no difference in vase life when compared to untreated flowers. However,
AOA and ASA had a slight effect in diminishing the initial rate of flower
abscission, which was not enough to extend the flower longevity (Finger
et al., 2004). In another work with roses, Brecheisen et al. (1995) found that
keeping the flowers in a solution containing 1% ASA significantly reduced
longevity when compared with tap water alone. In Consolida ajacis, neither
AOA nor ASA treatment was able to reduce the ethylene production of the
flowers, which might explain the lack of effect on the longevity.

3.4.2  Inhibition of Ethylene Action

The action of ethylene is efficiently inhibited by several inhibitors, including
silver ion, 1-methylcyclopropene (1-MCP), 2,5-norbornadiene, and nitrous
oxide. Among these inhibitors, 1-MCP seems to be the most promising
in blocking the ethylene, mainly due to its high affinity with the ethylene
receptor. Such ability makes the 1-MCP very effective at low concentra-
tions, ranging from 0.2 to 1 µl L −1. The commercial product is registered
by the name of EthylBloc ® for use in ornamental plants or SmartFresh® by
AgroFresh, Inc. 1-MCP is a worldwide commercialized growth plant regu-
lator, and it was classified as a nontoxic substance because similar natural
compounds are found in the atmosphere. 1-MCP is active at very low con-
centrations (Blankenship and Dole, 2003; PAN Pesticides Database, 2004).
Much work has been done in both ethylene-sensitive and -insensitive
flowers with 1-MCP. Usually the product is used just after harvest, fumigat-
ing the plants for 6–12 hours at room temperature, with different results
regarding its efficiency in blocking the ethylene action. The results vary
due to several endogenous and exogenous factors, including time between
harvest and the actual treatment, stage of development of the flower, con-
centration, and length of and temperature during the treatment.

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Cut flowers of Consolida ajacis treated with 1-MCP had their longevity
extended significantly, even when the flowers were exposed to 100 mg L −1
ethephon after they had been exposed to 0.5 g m−3 SmartFresh for 6 h at room
temperature. When ethephon was applied 2 h before the use of 1-MCP, lower
longevity was observed than with 1-MCP alone or when 1-MCP was applied
after the flowers had been treated with ethephon. But for ethephon-treated
flower, a longer vase life was observed than for control fl
­ owers (Table 3.5).
Thus, the reduction in flower longevity when ethephon was applied before
1-MCP suggests that the latter did not occupy the entire available receptor
sites for ethylene.
In another work, Cameron and Reid (2001) found that 1-MCP had
a transitory effect to block the abscission of Pelargonium peltatum petals
induced by ethylene, and a second fumigation was needed to prevent flower
abscission. Thus, the reduction of flower longevity when ethephon was
applied before 1-MCP in Consolida ajacis and the transient effect observed
in P. peltatum flowers suggest that binding sites for ethylene were not com-
pletely occupied by 1-MCP, or new receptors sites were available by the
presence of exogenous ethylene. It is well known that the presence of ethyl-
ene is able to make the flowers more sensitive to its action.
In addition to 1-MCP, other similar cyclopropene compounds have
shown high effectiveness in blocking ethylene action. 1-Hexylcyclopropene
(1-HCP) and 1-octylcyclopropene (1-OCP) more effectively blocked the del-
eterious effects of ethylene in Kalanchoë blossfeldiana cv. ‘Alexandra’ flow-
ers than 1-MCP (Kebenei et al., 2003). The authors found that pretreatment
with 1-OCP was more effective than that with 1-MCP in delaying flower
senescence at a concentration of 200 nl L −1 for 6 hours, followed by exposi-
tion to 2 µl L −1 ethylene, while the 1-HCP pretreatment had an effect similar
to that of 1-MCP, but at a 5 or 10 times higher concentration.

Table 3.5  Effect of Ethephon and 1-MCP on the Longevity of Consolida

ajacis Flowers
Treatments Longevity (days)
Control 4.5 c
Ethephon 1.4 d
1-MCP 6.0 a
Ethephon + 1-MCP 5.2 b
1-MCP + Ethephon 6.0 a
Source: Santos, V.R. et al., Bragantia 64, 33–38, 2005.
Longevity: Days for 50% of flower abscission or wilting. Mean separation within
columns by Tukey’s test; values followed by a letter in common are ­similar at the
5% level.

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Postharvest Quality of Ornamental Pl ants

Silver ion (Ag+) acts as an inhibitor of ethylene action. It is usually

sprayed or used in pulsing or in vase solutions to block the damaging
effects of ethylene, especially in flowers sensitive to ethylene. However, in
some flowers classified as having low or no sensitivity to ethylene, silver
ion may show beneficial effects when treated with silver thiosulfate (STS)
or AgNO3. In general, STS is more stable and has higher mobility within
the plant tissues than AgNO3; as a consequence, STS is considered less
phytotoxic and more effective at lower concentrations. STS is obtained by
mixing AgNO3 with sodium thiosulfate, with final concentrations varying
from 0.5 to 2.0 mM. To prepare a 0.1 M sodium thiosulfate stock solution,
mix 15.8 g of sodium thiosulfate into 500 ml of water. Separately, prepare
a 0.1 M silver nitrate stock solution by dissolving 16.9 g of silver nitrate
into 500 ml of water, keeping a 1:4 ratio of silver to thiosulfate. Pour the
silver nitrate solution onto the sodium thiosulfate under agitation. The STS
solution can be stored up to 1 month in the dark under refrigeration until
needed. Afterwards, the STS solution will lose its action due to silver pre-
cipitation and a new solution must be prepared.
Silver ion, mainly applied in the form of STS solution, has been used
commercially to prevent ethylene-induced senescence mainly in carnation.
But since silver is a heavy metal and considered a potential pollutant sub-
stance, other products have been tested to substitute Ag+ in the floriculture
industry. 1-MCP has been used as the most promising substitute of silver;
however, on some occasions STS is more efficient in extending the longev-
ity of ornamental crops than 1-MCP, in particular when the flowers are
stored in the absence of exogenous ethylene (Blankenship and Dole, 2003).
Pulsing roses with 1 mM AgNO3 or 1 mM STS for 3 hours delayed the petal
withering by 5–6 days compared to control flowers and extended the vase
life by 2.5 and 2.8 days, respectively (Son et al., 2003). Furthermore, due
to its action as a germicide at the base of the rose stem, silver ion was able
to maintain water potential at the flower peduncle throughout the vase life,
which resulted in improved water uptake by the stem. The treatment of
low ethylene-sensitive or ethylene-insensitive flowers does not always have
positive effects in extending their vase life. In tulips, for example, silver ion
showed no effect on the time, from the beginning of petal abscission, when
treated continuously with 0.5 mM STS (Sexton et al., 2000).

3.4.3  Ethylene Absorbers

Ethylene is an odorless and colorless gas that acts as a growth regulator
and harms horticultural crops. As an air pollutant, ethylene shows deleteri-
ous effects in ornamental plants in greenhouses, cold storage warehouses,
and inside display areas. Ethylene becomes active when the concentrations
in the atmosphere range from 0.1 to 1.0 µl L −1, and saturated responses

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occur between the concentrations of 1 and 10 µl L −1. In order to lower the

presence of ethylene in the air, absorbers can be placed in the storage
room and in trucks during shipping. In addition to the ethylene originating
from the senescence of fruits, flowers, and leaves, ethylene exhausts come
from combustion engines, cigarette smoke, leaky gas lines, and biomass
burning.
The most common absorbers are alumina and potassium permanga-
nate, with the latter being largely used as an active ingredient in filters
and sachets. Potassium permanganate is able to oxidize ethylene, form-
ing carbon dioxide and water, and is active in cold storage warehouses,
refrigerated trucks, and at noncontrolled room temperatures. As an inac-
tive ingredient, it can be used in pieces of florist’s foam, dry silica gel, or
vermiculite. One can make a saturated solution of potassium permanganate
and mix it with the inactive ingredient. If necessary, allow it to dry in an
oven at 70°C; consider replacing the filter or sachet when the color changes
from purple to brown.

3.5  Role of Abscisic Acid, Gibberellins,

and Cytokinins
The use of abscisic acid, gibberellins, and cytokinins as flower preserva-
tives is still limited, because not too many flowers respond with positive
effects and because of the higher cost of the substances than of other com-
mercial compounds regularly applied to cut flowers.
Benzyladenine (BA) had showed different effects depending on the
species and cultivar of flower used. When tropical flowers were sprayed or
dipped in a solution containing 200 mg L −1 from 10 s to 1 min, it prompted
no improvement on the vase life of Strelitzia reginae and Zingiber spectabi-
lis (Paull and Chantrachit, 2001). Nonetheless, the BA improved the vase
life of Anthurium andraeanum, Heliconia psitacorum, Heliconia chartacea,
and Alpinea purpurata flowers from 1.5- to 2.5-fold. In a similar study,
Moraes et al. (2005) found that the response of Heliconia latispatha to spray
with BA was dependent on the concentration. The improvement on the
vase life increased linearly when the flowers were sprayed with 100, 200,
or 300 mg L −1 BA, and at highest concentration, the gain in longevity was
1.85-fold superior to that of the untreated flowers. Transgenic petunia flow-
ers overexpressing the isopentenyl transferase (ipt) gene from the cytoki-
nin biosynthesis had an improved longevity of 6 to 10 days longer than the
wild-type flowers (Chang et al., 2003). Also in this same study, the authors
found that the transformed plants were less responsive to exogenous eth-
ylene in inducing flower wilting, showing that the increase in cytokinin
­content caused by the overproduction of the ipt gene reduced the ­sensitivity
of the flowers to the ethylene.

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Gibberellic acid (GA 3) was used in ornamental plants as an antagonist

to ethylene action, but its use as a preservative substance has shown other
effects in addition to reducing the flower senescence rate. Roses sprayed
with 1 mM GA 3 had suppressed the development of postharvest disease
caused by Botrytis cinerea by inhibiting the senescence-related changes in
the membrane permeability and protein degradation (Shaul et al., 1995).
The presence of exogenous ABA accelerated the leaf and flower
senescence of roses and carnation. When the daylily (Hemerocallis hybrid),
which is considered insensitive to ethylene, was treated with 100 µM ABA,
it caused premature accumulation of peroxidized lipids and induced the
activity of proteinase and RNAase, similar to the naturally occurring senes-
cence of the flower (Panavas et al., 1998). The authors suggest that ABA
plays an important role in the signal transduction events responsible for
the programmed death of daylily petal cells. In roses, Pompodakis et al.
(2004) found that the effect of ABA on flower longevity was dependent on
the pH used in the vase solution. In this work, the addition of 10 −5 M ABA at
pH 6 increased the vase life by inducing stomata closure in the presence or
absence of 1 mg L −1 AgNO3 as an antibacterial agent. But at alkaline pH 8, a
reduction of the vase life was observed, indicating that the beneficial effect
of ABA was eliminated by the microorganism growth present at high pH.

3.6  Role of Calcium on Flower Senescence

Pre- and postharvest applications of calcium have been applied to fresh
plant products to delay aging and postharvest decay and to control dis-
eases and physiological disorders. Flower and leaf senescence is delayed
by calcium treatment either before or after harvest, but the full mechanism
of its action still remains unclear. It is well known that calcium binds to
membrane phospholipids, protecting the membrane lipids from degrada-
tion, keeping the tissue function (Céour et al., 1992). Calcium also forms
Ca-pectates by cross-linking with COO− from the polygalacturonic acid at
medium lamella, increasing the rigidity of the plant cell wall. But the effec-
tiveness of calcium to improve the longevity of flowers such as cut roses
varies with the amount of calcium applied in the vase solution and also
by cultivar (Torre et al., 1999). In this work, the authors found that vase
solutions containing 1 mM CaCl 2 for the cv. ‘Baroness’ and 5 mM for the
cv. ‘Mercedes’ had delayed petal senescence, improved flower longevity,
enhanced opening of flower buds, postponed leakage of electrolytes, and
suppressed ethylene production during senescence. In addition, the CaCl 2
was able to keep the flower stems turgid for a longer period of time, retard-
ing the petal wilting when compared with untreated roses.
It has been observed that calcium also influences the development
of latent postharvest diseases in several fruits, vegetables, and flowers.

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When cv. ‘Kiss’ roses were sprayed with 10 or 20 mM calcium sulfate mixed
with 0.01% Tween 20 until the solution ran off, applied 24 hours before har-
vest, postharvest infection with Botrytis cinerea was decreased during
display, thus increasing the vase life by at least 30% at room temperature
(Capdeville et al., 2005). In a similar work, Capdeville et al. (2003) found
that pulsing the same rose cultivar with 50 mM calcium sulfate for 15 hours
reduced the severity of gray mold caused by Botrytis cinerea by 88% and
increase the vase life.

3.7 Respiration
In general, flowers have high respiration rates compared to other horticul-
tural products, which might lead to carbohydrate depletion because flowers
are not adapted as a long-term storage organ. In addition, other deleterious
effects develop under high respiratory activity, including higher transpira-
tion rates and elevated ethylene production and action. The Q10 factor for
most of vegetables and fruits is close to 2, but in flowers, it might reach
values up to 8 (Wills et al., 1998). In cut Narcissus flowers, the respiration
rate in two commercial cultivars was exponentially increased when the
temperature was increased from 0°C to 12.5°C, given a Q10 close to 3.5.
The elevation of the temperature resulted in a negative linear relationship
between the respiratory activity and the vase life (Cevallos and Reid, 2000).
Similar respiration and vase life behavior was observed for cut flowers of
gerbera and sunflower when kept under temperatures ranging from 0°C to
20°C (Çelikel and Reid, 2002). Thus, for these flowers, at the temperature
conditions studied, the respiration can be used as an excellent index to pre-
dict the longevity of the vase life.
The respiration of flowers with high sensitivity to ethylene can be
reduced by applying inhibitors of its action, in particular STS, which acts
as a persistent inhibitor of ethylene action. Altman and Solomos (1995)
found that carnation flowers had no response to exogenous ethylene
regarding the development of petal wilting or an increase in respiration
when the vase solution contained 0.2 mM STS. The senescence of the
flowers in Consolida ajacis is associated with the increase in ethylene pro-
duction and respiration (Finger et al., 2004). When cut flowers of C. ajacis
were pulsed with 1 mM STS or 1 mM STS + 5% sucrose, the increase
of respiration was inhibited, thus prolonging the vase life of the flower
(Figure 3.4). However, by loading the flowers with sucrose alone, no influ-
ence on the longevity was detected, probably due to the weak effect of the
carbohydrate in lowering the respiration compared to the STS (Finger
et al., 2004). Similarly to sucrose pulsing, spraying the flowers with 2 mM
AOA or 20 mM ASA had no effect on the respiration or, as a consequence,
the flower longevity (Figure 3.4).

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Postharvest Quality of Ornamental Pl ants

240 Control
5% sucrose
1mM STS
200 1 mM STS + 5% sucrose
AOA
ASA
160
Respiration (μl/g/h)

120

80

40

0
0 2 4 6 8 10 12 14 16
Days after harvest

Figure 3.4  Influence of pulsing with sucrose and STS and of spray with
AOA or ASA on respiration of Consolida ajacis flowers. (From Finger, F.L.
et al., Pesquisa Agropecuaria Brasileira 39, 533–537, 2004.)

3.8 Temperature
Temperature is the most important postharvest factor influencing the ­quality
and longevity of cut flowers and potted ornamental plants. Similar to any
fresh horticultural product, ornamental plants maintain high respiratory and
transpiratory activities to maintain their vital cell reactions. The intermedi-
ate organic molecules are used for the synthesis of new compounds and to
generate energy in the form of ATP. But respiration also produces heat as
a by-product, which increases when the temperature rises and accelerates
the aging process. The rate of senescence is dramatically reduced by cooling
the ornamental plants immediately after harvest. By far, the storage of cut
flowers at low temperatures has been used as the most adequate technique
for long-distance transport and, in many instances, for short-term storage
by the retail florists. The major effects of temperature on ornamental plant
storability and their posterior display are determined by the rate of respira-
tory substrate depletion and by the intensity of the transpiration rate and how
much production and action of ethylene will develop in the plants submitted
to temperature stress. Furthermore, all the previous effects of undesirable
storage temperatures will favor the development of postharvest diseases.

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In potted roses, especially in those cultivars sensitive to ethylene,

the dropping of the bud flowers and yellowing of the leaves are extremely
reduced when the pots are transported or stored around 5°C and pretreated
with STS spray or 1-MCP fumigation. Storage at below room tempera-
ture and under low-irradiance inside light exposure, somewhere around
5–15  µmol m−2 s −1, induces rapid senescence in potted roses, the effects
of which can be minimized by lowering the temperature and blocking the
ethylene action. A similar influence of temperature and light intensity is
observed in chrysanthemum, petunia, and geranium potted plants.
Tropical and subtropical flowers are sensitive to chilling tempera-
tures; such species cannot be stored below a critical temperature ranging
from 0°C to 13°C. The development of chilling symptoms is dependent on
the species or variety sensitivity, stage of development, length of storage
under the chilling-inducing temperatures, and the actual temperature of
storage. For most horticultural products, temperatures near 5°C are usu-
ally more effective in developing chilling symptoms. Several commercially
important cut flowers are sensitive to chilling, such as bird-of-paradise,
orchids, heliconias, ginger, and Alpinea purpurata, among other ornamen-
tal species.
Several symptoms of chilling injury develop in ornamental plants,
making the product unsuitable for commercialization. One of the first symp-
toms to develop is an increase of metabolite leakage in the cells, indicating
the lack of semipermeability in the cell membranes. As a consequence, the
leakage of metabolites facilitates the growth of microorganisms, especially
of fungi-related diseases. Other visual symptoms and metabolic imbalance
reactions will develop when plants are under chilling injury–inducing tem-
peratures. Some of the most common symptoms include leaf and flower
wilt, intense water loss, water-soaked tissues, and leaf and flower discol-
orations. As a result of a metabolic imbalance reaction, there is acceler-
ated senescence and ethylene production and accumulation of ethanol and
acetaldehyde.
The sensitivity and influence of storage length on the development
of chilling injury, and the subsequent influence on the vase life, were eval-
uated in bird-of-paradise flowers pulsed or not loaded with 40% sucrose
before storage at 10°C (Finger et al., 2003). Regardless of whether the
flowers were pulsed with sucrose before or after the cold storage, the
vase life at room temperature was reduced, with an increase in the length
of storage from 7 to 28 days (Table 3.6). In addition, the length of stor-
age reduced the total number of open florets at the end of the vase life
(Table  3.7). The most effective treatment in extending the flower lon-
gevity was achieved when the stems were pulsed after the cold storage
regardless of the number of weeks at 10°C, but when the flowers were
pulsed just before the cold storage, a fast decrease in poststorage vase

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Table 3.6  Vase Life of Bird-of-Paradise Flowers after Pulsing with 40%
Sucrose for 24 Hours and 7, 14, 21, and 28 Days of Storage at 10°C
Vase Life (Days)
Treatments/Days of Storage
at 10°C 7 14 21 28
Distilled water 8.3 a 6.7 b 3.3 a 1.6 a
Pulsed before cold storage 5.6 b 4.2 c 2.0 b 0.7 b
Pulsed after cold storage 8.6 a 8.0 a 4.0 a 2.0 a
Source: Finger, F.L. et al., Acta Horticulturae 628, 863–867, 2003.
Note: Mean separation within columns by Tukey’s test; values followed by a
­letter in common are not significantly different at the 5.0% level.

Table 3.7  Number of Open Florets of Bird-of-Paradise Flowers after Pulsing

with 40% Sucrose for 24 Hours and 7, 14, 21, and 28 Days of Storage at 10°C
Number of Open Florets
Treatments/Days of Storage
at 10°C 7 14 21 28
Distilled water 1.7 b 1.6 b 1.2 a 0.5 b
Pulsed before cold storage 1.4 b 1.5 b 1.1 a 1.0 a
Pulsed after cold storage 2.5 a 2.0 a 1.2 a 0.7 ab
Source: Finger, F.L. et al., Acta Horticulturae 628, 863–867, 2003.
Note: Mean separation within columns by Tukey’s test; values followed by a
­letter in common are not significantly different at the 5.0% level.

life was observed (Table 3.6). Furthermore, by pulsing the flowers with
40% sucrose for 24 h after the 10°C storage, a higher number of open
florets was obtained, at least after 7 and 10 days of storage (Table 3.7).
The reduced vase life after 28 days of storage was associated with the
development of chilling symptoms, characterized by discoloration and
Penicillium spp. contamination in the flowers.
When the storage temperature for a particular ornamental species
is not known, determine the optimum temperature of storage by placing
the flowers at different temperatures, usually ranging from 0°C up to 15°C.
Most of the temperate ornamental crops are chilling insensitive and can be
stored between 0°C and 2°C. However, for plants that originate from tropi-
cal and subtropical regions, it is always important to consider their sensitiv-
ity to chilling injury. In the majority of cases, the symptoms of injury will
develop under the chilling-induced temperatures; if not, the plants have to
be moved to a higher temperature in order to show any disturbance.

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3.9  Handling of Cut Flowers

In order to maintain the quality of cut flowers, it is necessary to address
the major factors that contribute to inducing or hastening the postharvest
deterioration during shipping, storage, and display. Understanding the sev-
eral factors that are directly involved in flower longevity will allow workers,
shippers, and wholesale and retail personnel to implement techniques to
maintain the quality of cut flowers:

1. Flower maturity: When flowers are harvested at their optimum

stage of development, a longer postharvest life will be achieved.
2. Stem cuts: Cut the stems with sharp and clean instruments and
immediately place them in buckets of water.
3. At the field: Keep the buckets with flowers in the shade and move
them to the packing house as soon as possible.
4. Temperature: After grading and cleaning the flowers, immediately
cool them to reduce the consumption of organic reserves, transpi-
ration and synthesis, and action of ethylene. If the flower is not
susceptible to chilling injury, cool the flowers to 2°C–4°C until
shipping or the product is moved to the display area.
5. Surface area: Flowers have a large surface area, which makes
them susceptible to intense water loss. To minimize wilting, store
the flowers in a cool chamber with relative humidity close to 95% or
cover them with perforated plastic bags.
6. Water: Use clean water at all times, and change it at least every
24 hours to avoid the growth of bacteria.
7. Buckets: Use light-colored buckets to make it easier to spot any
remaining dirt after cleaning.
8. Clean buckets: Wash the buckets with water containing 5% hypo-
chlorite before placing the flowers in them.
9. Hard water: Avoid the use of hard water with an alkaline pH in the
buckets. Use citric acid to lower the pH.
10. Tap water: Avoid the use of tap water due to the presence of fluo-
ride that can be toxic to some flowers, including roses and gladi-
olus. Instead of tap water, use rain water to keep the flowers.
11. Effects of ethylene: To reduce the undesirable effects of ethylene
on sensitive plants, the cold storage room should contain ethylene
absorber filters close to the evaporator fans. In cardboard boxes,
the use of ethylene absorber sachets can be effective in protecting
against ethylene action during shipping and storage.
12. Clean dying plant material: Clean up dying plant materials stored
in the cold storage chambers.
13. Moving methods: Avoid the use of gasoline- and propane-powered
carts around the storage areas.

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Postharvest Quality of Ornamental Pl ants

14. Clear flame: Make sure that any burner close to the storage room
has a clear flame because a yellow-flame burn releases high
­ethylene-concentrated fumes.
15. Stability: Avoid excessive transit in the cold storage room due to
rapid changes in the temperature and relative humidity.

3.10  Potted Plants

The center of origin of the genus Capsicum is America, although it is cur-
rently grown in tropical and temperate regions of the planet (Casali and
Couto, 1984). The Capsicum peppers grown in pots have a dual use as an
ornamental plant and a source of edible fruits. The dual use of ornamental
plants as a source of beauty and as food adds value to the product, increas-
ing the financial return of the producer (Finger et al., 2012).
The proper choice of cultivar for the production of potted peppers is
vital for success in this type of venture. In general, producers of ornamen-
tal chili peppers use plants with smaller and more colorful fruits. Thus,
for the production of peppers in small pots, growers should seek cultivars
that result in low-growing plants when the fruits are ripe and colorful fruit
with intense and upright green foliage without symptoms of early senes-
cence and abscission of leaves and fruit. Two popular cultivars are the
‘Pyramid’ pepper (C. frutescens) and ‘Calypso’ (C. annuum). The variet-
ies used in pots can be of any species of the genus Capsicum, but with a
predominance of C. annuum, due to the great diversity of its form and the
color of its fruit.
There are several factors that affect the longevity of ornamental pep-
pers when transferred to indoor environments where there is a deficiency
of light and water, as well as an accumulation of ethylene. Preliminary stud-
ies have shown that ethylene triggers a series of negative reactions in pot-
ted peppers, among them the abscission of the fruit, leaves, and flowers as a
reaction to the growing sensitivity to gas. However, other effects are visible,
such as the accelerated degradation of chlorophyll and flower senescence.
The ethylene concentration required to cause these effects is dependent
on factors such as exposure time, temperature, stage of development, and
sensitivity of species or variety (Hoyer, 1996).
The ethylene synthesis can be stimulated by various environmen-
tal factors, such as temperature extremes, mechanical injuries and disor-
ders, the composition of the storage atmosphere, drought, and disease.
According to Khan (2006), the ethylene response may vary among species,
and each part of the plant has a different level of sensitivity to this hormone.
The most suitable inhibitors of ethylene action belong to the cycloolefin
class, which can be used for consumption. Among the cycloolefins, 1-MCP
has been widely used in the treatment of fruits, vegetables, and ornamental

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Figure 3.5  Effect of ethylene and 1-MCP on the senescence of ornamental

pepper cultivar ‘Calypso’ (Capsicum annuum).

plants for control of fruit ripening and senescence of vegetables, cut ­flowers,
and ornamental potted plants (Blankenship and Dole, 2003).
Treatment with 1 g m−3 EthylBloc (0.14% of 1-MCP) for 6 h at a tem-
perature of 20°C to 25°C inhibited the action of ethylene on the abscission
of leaves of the ornamental pepper cultivar ‘Calypso’ (Segatto et al., 2013).
The application of 1-MCP extended the vase life of plants and completely
blocked the sites of ethylene action. Thus, once sprayed with 1-MCP, plants
do not exhibit abscission of leaves when exposed to ethylene (Figure 3.5).
There was no degradation of chlorophyll or leaf abscission in plants of the
cultivar ‘Calypso’, which were pretreated for 6 hours with 1 g m−3 EthylBloc
and subsequently exposed to an atmosphere containing 10 ml L −1 ethylene
at 48 h (Figure 3.5).

3.11  Conclusions and Future Perspectives

The longevity of flower and potted ornamental plants is determined by
the interaction of environmental, physiological, and genetic factors.
Water ­status, availability of carbohydrates, and ethylene action are deter-
minant factors in the development of senescence symptoms in most cut
flowers. In cut flowers and potted plants sensitive to ethylene, inhibitors

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Postharvest Quality of Ornamental Pl ants

of ethylene action prolong vase and shelf life by reducing flower wilting
and abscission of leaves, flowers, and fruits. Furthermore, inhibitors of
ethylene synthesis seem to be less effective than inhibitors of its action
in delaying flower and leaf senescence. Additional data are required to
establish the critical levels of ethylene for the senescence of many orna-
mental plants. The role of abscisic acid, gibberellins, and cytokinins in
the senescence of flowers must be better understood before they can be
of any practical use.

Acknowledgment
FAPEMIG and CNPq provided scholarship and financial support.

References
Altman, S.A., and Solomos, T. 1995. Differential and morphological responses
of carnations pulsed or continuously treated with silver thiosulfate.
Postharvest Biology and Technology 5, 331–343.
Blankenship, S.M., and Dole, J.M. 2003. 1-Methylcyclopropene: A review.
Postharvest Biology and Technology 28, 1–25.
Bleeksma, H.C., and Van Doorn, W.G. 2003. Embolism in rose stems as
a result of vascular occlusion by bacteria. Postharvest Biology and
Technology 29, 334–340.
Brecheisen, S., Haas, H.P., and Röber, R. 1995. Influence of water quality and
chemical compounds on vase life of cut roses. Acta Horticulturae 405,
392–400.
Cameron, A.C., and Reid, M.S. 2001. 1-MCP blocks ethylene-induced
petal abscission of Pelargonium peltatum but the effect is transient.
Postharvest Biology and Technology 22, 169–177.
Campanha, M.M., Finger, F.L., Cecon, P.R., and Barbosa, J.G. 1997. Water
relations of cut bird-of-paradise (Strelitzia reginae Ait.) inflorescences.
Revista Brasileira Horticultura Ornamental 3, 27–31.
Capdeville, G. de, Maffia, L.A., Finger, F.L., and Batista, U.G. 2003. Gray mold
severity and vase life of rose buds after pulsing with citric acid, sali-
cylic acid, calcium sulfate, sucrose and silver thiosulfate. Fitopatologia
Brasileira 28, 380–385.
Capdeville, G.D., Maffia, L.A., Finger, F.L., and Batista, U.G. 2005. Pre-harvest
calcium sulfate applications affect vase life and severity of gray mold in
cut roses. Scientia Horticulturae 103, 329–338.
Carneiro, T.F., Finger, F.L., Santos, V.R., Neves, L.L.M., and Barbosa, J.G.
2002. Influência da sacarose e da base da haste na longevidade de
inflorescências de Zinnia elegans. Pesquisa Agropecuaria Brasileira 37,
1065–1070.

105
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Casali, V.W.D., and Couto, F.A.A. 1984. Origem e botânica de Capsicum.

Informe Agropec 10, 8–10.
Çelikel, F.G., and Reid, M.S. 2002. Storage temperature affects the quality of
cut flowers from the Asteraceae. HortScience 37, 148–150.
Céour, F., Arul, J., Makhlouf, J., and Willemot, C. 1992. Delay of membrane
lipid degradation by calcium treatment during cabbage leaf senescence.
Plant Physiology 100, 1656–1660.
Cevallos, J.C., and Reid, M.S. 2000. Effects of temperature on the respiration
and vase life of Narcissus flowers. Acta Horticulturae 517, 335–341.
Chang, H., Jones, M.L., Banowetz, G.M., and Clark, D.G. 2003.
Overproduction of cytokinins in petunia flowers transformed with
PSAG12-IPT delays corolla senescence and decreases sensitivity to ethyl-
ene. Plant Physiology 132, 2174–2183.
Fan, X., Mattheis, J.P., and Fellman, J.K. 1996. Inhibition of apple
fruit  1-aminocyclopropane-1-carcoxylic acid oxidase and respiration
by ­acetylsalicylic acid. Journal of Plant Physiology 149, 469–471.
Finger, F.L., Carneiro, T.F., and Barbosa, J.G. 2004. Senescência pós-­colheita
de flores de esporinha (Consolida ajacis). Pesquisa Agropecuaria
Brasileira 39, 533–537.
Finger, F.L., Moraes, P.J., Barbosa, J.G., and Grossi, J.A.S. 2003. Vase life of
bird-of-paradise flowers influence by pulsing and term of cold storage.
Acta Horticulturae 628, 863–867.
Finger, L.F., Rêgo, E.R, Segatto, F.B., Nascimento, N.F.F., and Rêgo, M.M.
2012. Produção e potencial de mercado para pimenta ornamental.
Informatica Agropecuaria 33, 14–20.
Hoyer, L. 1996. Critical ethylene exposure for Capsicum annuum ‘Janne’ is
dependent on an interaction between concentration, duration and devel-
opmental stage. Journal of Horticultural Science 71, 621–628.
Ichimura, K., Kojima, K., and Goto, R. 1999. Effects of temperature,
8-hydroxyquinoline sulfate and sucrose on the vase life of cut rose flow-
ers. Postharvest Biology and Technology 15, 33–40.
Kebenei, Z., Sisler, E.C., Winkelmann, T., and Serek, M. 2003. Efficacy of
new inhibitors of ethylene perception in improvement of display life of
kalanchoë (Kalanchoë blossfeldiana Poelln.) flowers. Postharvest Biology
and Technology 30, 169–176.
Ketsa, S., and Narkbua, N. 2001. Effect of aminooxiacetic acid and sucrose on
vase life of cut roses. Acta Horticulturae 543, 227–234.
Khan, A.N. 2006. Ethylene Action in Plants. Springer, Amsterdam, Netherlands.
Liao, L.J., Lin, Y.H., Huang, K.L., and Chen, W.S. 2001. Vase life of Eustoma
grandiflorum as affected by aluminum sulfate. Botanical Bulletin of
Academia Sinica 42, 35–38.
Mapeli, A.M., Finger, F.L., Oliveira, L.S., and Barbosa, J.G. 2009. Longevidade
de inflorescências de Epidendrum ibaguense tratadas com aminoetoxi-
vinilglicina. Pesquisa Agropecuaria Brasileira 44, 258–262.

106
Postharvest Quality of Ornamental Pl ants

Moraes, P.J., Finger F.L., Mapeli A.M., Cecon P.R. and Barbosa J.G. 2009.
Growth and Flower Development of Epidendrum ibaguense Orchid.
Acta Horticulturae 813, 565–570.
Newman, J.P., Dodge, L.L., and Reid, M.S. 1998. Evaluation of ethylene
inhibitors for postharvest treatment of Gypsophila paniculata L.
HortTechnology 8, 58–63.
PAN Pesticides Database. 2004. www.pesticideinfo.org.
Panavas, T., Walker, E.L., and Rubinstein, B. 1998. Possible involvement of
abscisic acid in senescence of daylily petals. Journal of Experimental
Botany 49, 1987–1997.
Paull, R.E., and Chantrachit, T. 2001. Benzyladenine and the vase life of tropi-
cal ornamentals. Postharvest Biology and Technology 22, 303–310.
Pompodakis, N.E., Joyce, D.C., Terry, L.A., and Lydakis, D.E. 2004.
Effects of  vase solution pH and abscisic acid on the longevity of cut
‘Baccara’ roses. Journal of Horticultural Science and Biotechnology 79,
828–832.
Porat, R., Borochov, A., and Halevy, A.H. 1994a. Pollination-induced changes
in ethylene production and sensitivity to ethylene in cut dendrobium
orchid flowers. Scientia Horticulturae 58, 215–221.
Porat, R., Borochov, A., Halevy, A.H., and O’Neil, S.D. 1994b. Pollination-
induced senescence of Phalaenopsis petals. Plant Growth Regulation 15,
129–136.
Santos, V.R., Finger, F.L., Barbosa, J.G., and Barros, R.S. 2005. Influência do
etileno e do 1-MCP na senescência e longevidade de inflorescências de
esporinha. Bragantia 64, 33–38.
Segatto, F.B., Finger, F.L., Barbosa, J.G., Rêgo, E.R., and Pinto, C.M.F. 2013.
Effects of ethylene on the post-production of potted ornamental peppers
(Capsicum annuum L.). Acta Horticulturae 1000, 217–222.
Setyadjit, S., Joyce, D.C., Irving, D.E., and Simons, D.H. 2004. Development
and senescence of Grevillia ‘Sylvia’ inflorescences, flowers and flower
parts. Plant Growth Regulation 44, 133–146.
Sexton, R., Laird, G., and Van Doorn, W.G. 2000. Lack of ethylene involve-
ment in tulip tepal abscission. Physiologia Plantarum 108, 321–329.
Shaul, O., Elad, Y., and Zieslin, N. 1995. Suppression of Botrytis blight in
cut roses flowers with gibberellic acid: Effects of postharvest timing
of the gibberellin treatment, conidial inoculation and storage period.
Postharvest Biology and Technology 6, 331–339.
Son, K.C., Byoun, H.J., and Yoo, M.H. 2003. Effect of pulsing with AgNO3 or
STS on the absorption and distribution of silver and the vase life of cut
rose ‘Red Sandra’. Acta Horticulturae 624, 365–372.
Torre, S., Borochov, A., and Halevy, A.H. 1999. Calcium regulation of senes-
cence in rose petals. Physiologia Plantarum 107, 214–219.
Van Doorn, W.G. 1997. Water relations of cut flowers. Horticultural Reviews
18, 1–85.

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Van Doorn, W.G. 2002. Effect of ethylene on flower abscission: A survey.

Annals of Botany 89, 689–693.
Van Doorn, W.G., and Vaslier, N. 2002. Wounding-induced xylem occlusion in
stems of cut chrysanthemum flowers: Roles of peroxidase and cathecol
oxidase. Postharvest Biology and Technology 26, 275–284.
Van Doorn, W.G., Witte, Y., and Harkema, H. 1995. Effects of high number of
exogenous bacteria on the water relations and longevity of cut carnation
flowers. Postharvest Biology and Technology 6, 111–119.
Wills, R., McGlasson, B., Graham, D., and Joyce, D. 2008. Postharvest: An
Introduction to the Physiology and Handling of Fruit, Vegetables and
Ornamentals. 4th ed. CABI, Wallingford, UK.
Woltering, E.J., and Van Doorn, W.G. 1988. Role of ethylene in senescence
of petals—morphological and taxonomical relationships. Journal of
Experimental Botany 208, 1605–1616.

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Chapter 4

Physiology and
Molecular Biology of
Flower Senescence
Kenichi Shibuya and Kazuo Ichimura
National Agriculture and Food Research
Organization (NARO), Fujimoto, Tsukuba, Japan

Abstract 110
4.1 Introduction 110
4.2 Ethylene and Senescence of Cut Flowers 111
4.2.1 Ethylene Response of Cut Flowers 111
4.2.2 Types of Senescence in Cut Flowers with High
Ethylene Sensitivity 112
4.2.3 Ethylene in Petal-Wilting-Type Flowers 113
4.2.4 Ethylene in Petal-Abscission-Type Flowers 114
4.2.5 Ethylene Biosynthesis 114
4.2.6 Ethylene Signal Transduction 115
4.2.7 Acceleration of Flower Senescence by
Pollination 116
4.2.8 Acceleration of Flower Senescence by
Wounding 119
4.2.9 Effect of Temperature on Ethylene Production
and Perception 119

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4.3 Plant Hormones Other than Ethylene in Flower

Senescence 120
4.3.1 Auxin 120
4.3.2 Gibberellin 120
4.3.3 Cytokinin 120
4.3.4 Abscisic Acid 121
4.3.5 Jasmonic Acid 122
4.4 Programmed Cell Death in Flower Senescence 122
4.4.1 Programmed Cell Death 122
4.4.2 Gene Expression during PCD in Flowers 123
4.4.3 Autophagy in Petal Senescence 124
4.5 Conclusions and Future Perspectives 124
References 125

Abstract
Flower longevity is one of the most important traits of cut flowers. An under-
standing of the postharvest biology of flowers is needed to efficiently develop
postharvest technology. Ethylene is a key factor that regulates flower senes-
cence in many plant species. Ethylene biosynthetic and signaling pathways
have been well characterized, and it is now possible to improve the vase life
of some cut flowers by controlling the effects of ­ethylene. In contrast, regula-
tory mechanisms of senescence in flowers that show ­ethylene-independent
senescence remain largely unknown. Petal ­senescence is a highly regulated
developmental process and is considered to be a type of programmed cell
death, which is an actively regulated ­process. To identify key regulators of
programmed cell death during petal senescence, many studies have been
conducted. In this chapter, we outline studies on the postharvest physiology
and molecular biology of flowers, focusing on regulatory mechanisms for
petal senescence by ethylene and programmed cell death.

4.1 Introduction
Several postharvest techniques are applied to improve the vase life of many
cut flowers. These techniques have been developed based on knowledge
from basic studies of plant hormones, sugar, and water relations. Ethylene
plays a very important role in the senescence of many cut ­flowers, and this
plant hormone has been of primary interest in the postharvest biology of
flowers. Besides ethylene, other plant hormones play an important role in
flower senescence and the biochemical functions of biomembranes during
senescence. After the 1990s, with the advances in molecular b ­ iology, genes

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involved in ethylene biosynthesis and ethylene signal transduction were

revealed in many ornamental plants. More recently, gene expression has
been analyzed comprehensively in several flowers, and numerous senes-
cence-related genes have been isolated. On the other hand, many reports
on the water relations of cut flowers have advanced the knowledge of post-
harvest physiology. Several reviews on the postharvest biology of flowers
have been published (Rogers, 2006, 2013; Tripathi and Tuteja, 2007; Van
Doorn and Woltering, 2008; Ichimura, 2010; Shibuya, 2012). In this chapter,
we review studies on the postharvest physiology and molecular biology of
flowers, focusing on regulatory mechanisms for petal senescence.

4.2  Ethylene and Senescence of Cut Flowers

4.2.1  Ethylene Response of Cut Flowers

Ethylene accelerates petal wilting or abscission of flower parts in some
plant species but has little effect in other plant species. Sensitivity to ethyl-
ene in cut flowers is generally evaluated based on whether signs of senes-
cence are observed after exposure to a constant concentration of ethylene.
Woltering and Van Doorn (1988) evaluated ethylene sensitivity in 96 plant
species from 24 families by treating with 3 ppm ethylene for 22–24 hours.
They reported that flowers of plant species belonging to the Asteraceae and
Iridaceae exhibit lower sensitivity to ethylene, whereas flowers belonging to
the Caryophyllaceae exhibit higher sensitivity, suggesting that differences
in ethylene sensitivity may depend on genetic factors. However, more recent
works found that longer exposure to ethylene results in accelerated flower
senescence in some species. For example, daffodil was classified as a flower
with very low ethylene sensitivity, but continuous treatment with ethylene
hastens flower senescence, and treatment with ethylene inhibitors delays
senescence (Hunter et al., 2004b). In some plant species, ethylene sensitiv-
ity of flowers increases with aging. Sepal abscission of Delphinium hybrid
cv. ‘Bellamosum’ flowers is not induced by ethylene treatment (40 µl L −1)
on the day of harvest, but it is induced if flowers are treated with ethylene
1 day after harvest (Ichimura et al., 2009a). Thus, time of treatment, con-
centration of ethylene, and age of flowers need to be considered in order to
evaluate the sensitivity to ethylene in cut flowers.
Sensitivity to ethylene varies greatly among plant species. In carna-
tion, 0.6 µl L −1 ethylene induces petal wilting within 12 hours (Wu et al.,
1991), while in rose it takes more than 2 days until flowers abscise in continu-
ous 1 µl L −1 ethylene (Ichimura et al., 2005). In chrysanthemum, little effect
is observed even when flowers are treated with 1 µl L −1 ethylene for more
than 10 days (Doi et al., 2003). Here, we have classified ethylene sensitivity
of cut flowers based on observation of their responses to ethylene (Table 4.1).

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Table 4.1  Ethylene Sensitivity in Cut Flowers

Ethylene
Sensitivity Plant Species References
Very high Carnation (Nichols, 1968)
High Gypsophila paniculata (Woltering and Van Doorn, 1988)
Sweet pea (Shimizu-Yumoto and Ichimura, 2006a)
Delphinium (Ichimura et al., 2009a)
Dendrobium (Porat et al., 1994)
Vanda (Goh et al., 1985)
Relatively Campanula (Kato et al., 2002)
high Snapdragon (Ichimura et al., 2007)
Stock (Celikel and Reid, 2002)
Eustoma (Shimizu-Yumoto and Ichimura, 2009b)
Rose (Ichimura et al., 2005)
Blue star (Tweedia caerulea) (Hiraya et al., 2002)
Relatively Alstroemeria (Wagstaff et al., 2005)
low Daffodil (Hunter et al., 2004b)
Low Chrysanthemum (Doi et al., 2003)
Gladiolus (Serek et al., 1994)
Tulip (Sexton et al., 2000)
Lily (oriental hybrid) (Elgar et al., 1999)
Note:  Very high, petal wilting observed within 24 hours of treatment with ethylene
at less than 1 µl L−1; high, petal wilting or flower abscission observed within
24 hours of treatment with ethylene at 1–10 µl L−1; relatively high, petal wilt-
ing or flower abscission observed within 48 hours of treatment with ethylene
at 1–10 µl L−1; ­relatively low, acceleration of senescence symptoms eventually
observed at a significant level after ethylene or ethrel treatment; low, accelera-
tion of senescence symptoms not observed after ethylene treatment.

Some flowers, including Delphinium (Ichimura et al., 2009a), Eustoma

(Shimizu-Yumoto and Ichimura, 2009b), Portulaca (Ichimura and Suto,
1998), and Torenia (Goto et al., 1999), show an increase in ethylene sensitiv-
ity as flowers age. Intriguingly, ethylene sensitivity declines after harvest
in carnation flowers, which are highly sensitive to ethylene (Mayak and
Dilley, 1976b; Onozaki et al., 2004).

4.2.2  Types of Senescence in Cut Flowers

with High Ethylene Sensitivity
Senescence patterns of flowers that show high ethylene sensitivity can be
classified into two types: a petal-wilting type, in which ethylene induces
petal wilting, and a petal-abscission type, in which ethylene induces

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abscission of petals and sepals. Abscission of flower parts is regulated by

­ethylene in many plant species, although there are exceptions, such as tulip
(Sexton  et al., 2000). Petal-wilting-type flowers include carnation, sweet
pea, and orchids, among others. Delphinium is a representative of a petal-
or sepal-abscission-type flower. In flowers such as sweet pea and blue star
(Tweedia caerulea), petals or whole flowers abscise after petals have wilted.
In these cases, senescence is generally classified by type based on whether
wilting or abscission occurs first.

4.2.3  Ethylene in Petal-Wilting-Type Flowers

Petal-wilting-type flowers can be further classified into two groups based
on patterns of ethylene production during flower senescence. Carnation
(Nichols, 1966), sweet pea (Mor et al., 1984), and Eustoma (Ichimura and
Goto, 2000) exhibit a clear increase in ethylene production during natural
(unpollinated) senescence. In contrast, ethylene synthesis does not rise
during natural senescence in some flowers, including Campanula (Kato
et al., 2002). In flowers in which ethylene synthesis rises during natural
senescence, treatment with ethylene inhibitors such as STS (anionic silver
thiosulfate complex) delays flower senescence. On the other hand, in flow-
ers in which ethylene synthesis does not rise during natural senescence,
such as Campanula (Kato et al., 2002), ethylene inhibitors have little effect
on the progression of senescence. In Campanula, ethylene production is
induced by pollination, resulting in hastened senescence. In this case, STS
treatment can suppress the increase in ethylene production and cancel the
effect of pollination on senescence (Kato et al., 2002).
Ethylene synthesis rises as flowers senesce, and treatment with ethyl-
ene inhibitors delays petal wilting in some petal-wilting-type flowers, indicat-
ing that ethylene regulates petal wilting in these flowers. Ethylene production
induced by ethylene itself is called autocatalytic ethylene production. The
climacteric-like rise in ethylene production during petal senescence in many
petal-wilting-type flowers is the result of autocatalytic ethylene ­production.
Since petal wilting is accompanied by autocatalytic ethylene production,
these two events are thought to be closely linked and inseparable. However,
in transgenic carnation, in which expression of an ACC oxidase gene is
­suppressed, exogenous ethylene treatment induces petal wilting but not auto-
catalytic ethylene production, suggesting that these two events may be under
the control of different pathways (Kosugi et al., 2000).
Ethylene is produced from virtually all organs of flowers, but its
­production varies among organs. Even within the same organ, production
differs among parts of the organ. In carnation petals, for example, the base
of the petals produces more than 10-fold more ethylene than the tip of the
petals on a fresh weight basis (Mor et al., 1985).

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In carnation flowers, an increase in ethylene production starts in the

gynoecium prior to petals (ten Have and Woltering, 1997). Thus, gynoe-
cium-borne ethylene is thought to induce ethylene production in the pet-
als, leading to petal wilting. In fact, removal of the gynoecium has been
reported to suppress an increase in ethylene production in petals and to
dramatically delay petal senescence (Shibuya et al., 2000). However, the
role of the gynoecium in petal senescence needs to be clarified, since sev-
eral studies have reported different results (Mor et al., 1980; Sacalis and
Lee, 1987; Woodson and Brandt, 1991).
In Portulaca flowers, stamens produce more ethylene on a fresh
weight basis than petals or the gynoecium. When stamens are touched,
a large amount of ethylene is produced only from stamens, leading to
petal senescence (Shimizu and Ichimura, 2001). In contrast, wounding of
the gynoecium or petals does not accelerate petal senescence (Ichimura
and Suto, 1998). This suggests that in the case of Portulaca, the ethylene
­produced from stamens regulates petal senescence.

4.2.4  Ethylene in Petal-Abscission-Type Flowers

Petal-abscission-type flowers can be classified into two groups: flowers
that show autocatalytic ethylene production during senescence, such as
Delphinium (Ichimura et al., 2009a) and Torenia (Goto et al., 1999), and
flowers that do not show autocatalytic ethylene production during senes-
cence, such as geranium (Clark et al., 1997). In flowers of both types, an
increase in ethylene production is observed in tissues other than the petals.
In Delphinium flowers, ethylene is mainly produced from gynoe-
cia and receptacles, and its amount rapidly increases during senescence.
Wounding of gynoecia or receptacles induces ethylene production in these
organs and accelerates sepal abscission (Ichimura et al., 2009a). Since
sepals connect directly with receptacles in Delphinium, ethylene produced
from receptacles seems to play a major role in sepal abscission. In Digitalis
(Stead and Moore, 1979) and Torenia (Goto et al., 1999), ethylene produc-
tion rises in the gynoecium, while a little ethylene is produced in petals
during senescence. Thus, in these plants, ethylene that is produced from
the gynoecium is likely involved in petal abscission.

4.2.5  Ethylene Biosynthesis

The ethylene biosynthetic pathway in plants has been established, and
genes encoding key enzymes have been isolated (Yang and Hoffman, 1984;
Kende, 1993; Lin et al., 2009). Ethylene is synthesized through the follow-
ing pathway:

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l -Methionine → S-adenosyl-l -methionine → 1-aminocyclopropane-

1-carboxylic acid (ACC) → ethylene

The last two reactions are catalyzed by ACC synthase and ACC
oxidase.
In vegetative tissues of plants, ACC synthase is considered to be a
rate-limiting step in ethylene biosynthesis. However, activities of not only
ACC synthase but also ACC oxidase rise during petal senescence of car-
nation, suggesting that both enzymes are responsible for the increase in
ethylene production (Woodson et al., 1992). In the gynoecium of carnation
flowers, on the other hand, ACC oxidase activity is constantly high during
senescence, and thus the increase in ethylene production in this tissue is
likely due to a rise in ACC synthase activity (Yangkhamman et al., 2007).
ACC synthase and ACC oxidase are encoded by multigene families,
and genes encoding these enzymes have been isolated in many ornamental
plant species. Three ACC synthase genes have been isolated from carna-
tion (Park et al., 1992; Henskens et al., 1994; Jones and Woodson, 1999a),
four from rose (Wang et al., 2004), three from snapdragon (Woltering et al.,
2005), two from geranium (Wang and Arteca, 1995), and two from petunia
(Lindstrom et al., 1999). ACC synthase genes are differentially regulated
in a tissue-specific manner. For example, among the three carnation ACC
synthase genes, DcACS1 is most abundant in petals, while DcACS2 and
DcACS3 are preferentially expressed in styles (Jones and Woodson, 1999a).
For ACC oxidase, two genes have been isolated from carnation
(Wang and Woodson, 1991; Norikoshi et al., 2008), one from rose (Xue
et al., 2008), one from snapdragon (Woltering et al., 2005), one from
Phalaenopsis (Nadeau et al., 1993), five from tulip (Momonoi et al., 2007),
four from ­petunia (Tang et al., 1993), and three from geranium (Clark
et al., 1997). ACC oxidase genes are regulated in a spatial- and temporal-­
specific m­ anner. In carnation, DcACO1 is clearly upregulated in petals dur-
ing ­senescence, while DcACO2 is constantly expressed in the gynoecium
(Norikoshi et al., unpublished data). Differential expression of ACC oxi-
dase genes has also been reported in petunia (Tang et al., 1994).

4.2.6  Ethylene Signal Transduction

Ethylene signaling is mediated by a complex multicomponent pathway
(Lin et al., 2009). ETR1 (ETHYLENE RESPONSE1), which encodes a His
kinase with homology to bacterial two-component regulators, has been
identified as an ethylene receptor (Chang et al., 1993). Five ethylene recep-
tor genes have been cloned from Arabidopsis thaliana (Hua et al., 1995;
Sakai et al., 1998). Analysis of loss-of-function mutations in multiple recep-
tors has shown that these receptors are negative regulators of ethylene

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responses (Hua and Meyerowitz, 1998). The receptors act through CTR1
(CONSTITUTIVE TRIPLE RESPONSE1), which is homologous to the
Raf family of Ser/Thr kinases and negatively regulates ethylene signaling
(Kieber et al., 1993; Huang et al., 2003). Downstream of the receptor–CTR1
complex is EIN2 (ETHYLENE INSENSITIVE2), which has homology to
Nramp metal transporters and positively regulates signaling (Alonso et al.,
1999). EIN2 undergoes proteasome-mediated turnover (Qiao et al., 2009),
and ethylene stimulates phosphorylation-dependent cleavage and nuclear
movement of a carboxyl-terminal EIN2 fragment (Qiao et al., 2012). Toward
the end of the signaling pathway, ethylene responses are mediated by tran-
scription factors including EIN3 (ETHYLENE INSENSITIVE3) and ERF1
(ETHYLENE RESPONSE FACTOR1) (Chao et al., 1997; Solano et  al.,
1998). Regulation of the stability of EIN3 protein is a key aspect of the ethyl-
ene response (Guo and Ecker, 2003; Yanagisawa et al., 2003).
Genes encoding ethylene receptors have been isolated in several
ornamental plant species, for example, three in carnation (Shibuya et al.,
2002), five in rose (Müller et al., 2000a, 2000b), one in chrysanthemum
(Narumi et al., 2005), two in Delphinium (Kuroda et al., 2003; Tanase and
Ichimura, 2006), four in petunia (Wang and Kumar, 2007), two in geranium
(Dervinis et al., 2000), and two in gladiolus (Arora et al., 2006). Genes
have been reported that encode CTR1 in rose (Müller et al., 2002) and
Delphinium (Kuroda et al., 2004), EIN2 in petunia (Shibuya et al., 2004),
and EIN3 in carnation (Waki et al., 2001; Iordachescu and Verlinden, 2005),
rose (Müller et al., 2003), and petunia (Shibuya and Clark, 2006).
Ethylene receptors are negative regulators of ethylene signaling, and
depletion of the receptors is considered to lead to an increase in ethylene
sensitivity (Hua and Meyerowitz, 1998). However, it has been reported that
the transcript levels of ethylene receptors in rose are higher in cultivars with
higher ethylene sensitivity in rose (Müller et al., 2000a). The inconsistency of
the relationship between levels of transcript for ethylene receptors and ethyl-
ene sensitivity might be explained by posttranslational regulation of ethylene
receptor proteins. Kevany et al. (2007) showed that protein levels for ethylene
receptors in tomato are not correlated with corresponding transcript levels
during fruit development and that the receptors are rapidly degraded in the
presence of ethylene. Thus, the level of ethylene receptor proteins seems
to be responsible for ethylene sensitivity. Similar regulation of the ethylene
response is expected in flower senescence. Analysis of ethylene receptors at
the protein level is necessary in the study of flower senescence.

4.2.7  Acceleration of Flower Senescence by Pollination

Pollination accelerates petal wilting and abscission of flower parts in spe-
cies that show relatively high ethylene sensitivity, including orchids (Burg

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and Dijkman, 1967; Porat et al., 1994), petunia (Gilissen and Hoekstra,
1984; Whitehead et al., 1984), Eustoma (Ichimura et al., 1998; Ichimura
and Goto, 2000), carnation (Larsen et al., 1995; Jones and Woodson, 1997),
Campanula (Kato et al., 2002), Delphinium (Okamoto and Ichimura, 2010),
and Torenia (Goto et al., 1999). In particular, flower longevity of orchid spe-
cies is dramatically shortened by pollination. In daffodil flowers, which
show relatively low ethylene sensitivity, pollination also accelerates flower
senescence (Hunter et al., 2004b).
Pollination raises ethylene production from flowers, and treatment
with inhibitors of ethylene biosynthesis or ethylene perception cancels
the acceleration of flower senescence in many plant species (Stead, 1992).
Furthermore, transgenic petunia plants with reduced ethylene sensitiv-
ity due to suppressed expression of PhEIN2 exhibit significant delays in
petal senescence after pollination (Figure 4.1) (Shibuya et al., 2004). These
observations suggest that ethylene is a primary signal for pollination-­
accelerated flower senescence.
In Eustoma (Shimizu-Yumoto and Ichimura, 2006b), petunia (Gilissen,
1977), and Digitalis (Stead and Moore, 1979), a greater amount of pollen
­pollinating the stigma results in faster progression of flower senescence.
In  this case, a greater concentration of ethylene inhibitors is needed to
­counter the effect of pollination in flower senescence.
Compatibility between pollen and gynoecium involves the accel-
eration of flower senescence by pollination. In self-incompatible petunia,
compatible pollination accelerates the second peak of ethylene synthesis,
leading to hastened petal senescence, while incompatible pollination does

Figure 4.1  Petal senescence in transgenic petunia with reduced ethylene

sensitivity. Flower of a wild-type plant 2 days after pollination (left) and
flower of transgenic plant with reduced PhEIN2 expression 8 days after pol-
lination (right). The petals of transgenic plants are still turgid even after the
ovaries have fully expanded. (From Shibuya, K. et al., Plant Physiology 136,
2900–2912, 2004.)

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not accelerate the second peak of ethylene synthesis and does not lead to
hastened petal senescence (Singh et al., 1992). Similar phenomena have
been observed in carnation (Larsen et al., 1995).
In pollinated orchid flowers, ethylene sensitivity rises prior to an
increase in ethylene production. This rise in ethylene sensitivity is not
suppressed by an ethylene synthesis inhibitor, AOA, suggesting that the
increase in ethylene sensitivity is regulated independently from e­ thylene
synthesis (Porat et al., 1994, 1995a). In addition, treatment with STS, an eth-
ylene perception inhibitor, suppresses the increase in ethylene ­production.
These observations suggest that the dramatic rise in ­ethylene production
after pollination in orchid flowers may be caused by the perception of a rela-
tively low level of ethylene as a result of an increase in ethylene sensitivity.
Detailed analyses of ethylene production patterns after pollination
have been conducted in several flower species. Petunia flowers produce
ethylene in styles within 3 h and then in petals at 2–3 days after pollination
(Hoekstra and Weges, 1986; Singh et al., 1992). In carnation, an increase
in ethylene production is observed at around 12 hours in the stigma and at
24 hours in ovaries, and then at 36 hours in petals after pollination (Jones
and Woodson, 1999b). In Phalaenopsis, ethylene increases at 12 hours in the
stigma and column, at 24 hours in the labellum, and after 48 hours in the
perianth after pollination (O’neill et al., 1993).
It has been reported that in pollinated Phalaenopsis flowers, the accu-
mulation of mRNA for ACC synthase and ACC oxidase increases in the
gynoecium, while mRNA for ACC oxidase is not detectable in the perianth
(O’Neill et al., 1993). Based on this observation, the following model has
been proposed: in the perianth, ACC is translocated from other flower parts,
such as the gynoecium, and is converted to ethylene (O’Neill et al., 1993).
However, mRNA for ACC synthase has since been detected in the perianth
using real-time reverse transcription polymerase chain reaction (RT-PCR),
a more sensitive technique, and the level of PhalACS1 mRNA increases in
the perianth after pollination (Ichimura and Niki, unpublished data).
In petunia, when the style is removed within several hours of pol-
lination, petal senescence is not accelerated by pollination (Gilissen and
Hoekstra, 1984). This observation suggests the existence of mobile signals
that induce petal senescence after pollination. Candidates for the mobile
signals are ACC and ethylene. When radiolabeled ACC is applied to the
stigmas of carnation flowers, radioactive ethylene is produced both by the
gynoecia and by the petals, suggesting that ACC may be a signaling com-
pound transported from the stigmas to the petals (Reid et al., 1984). In con-
trast, radiolabeled ACC applied to the stigma is largely immobile in flowers
of Cymbidium (Woltering et al., 1995) and petunia (Woltering et al., 1997).
Furthermore, in petunia, eluates collected from the styles were able to
induce petal senescence, but ACC was not detected in the eluates (Gilissen
and Hoekstra, 1984). These observations suggest that ACC is not a mobile

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signaling factor for petal senescence in these plant species. In addition to

chemical compounds, electrical signaling has been proposed to be involved
in the induction of petal senescence after pollination (Spanjers, 1981;
Fromm et al., 1995).

4.2.8  Acceleration of Flower Senescence by Wounding

In plant species in which flower senescence is hastened by pollination,
wounding of gynoecia often accelerates flower senescence. Wounding of
the stigma results in accelerated petal senescence in Eustoma (Ichimura
and Goto, 2000) and petunia (Lovell et al., 1987; Whitehead et al., 1984),
and hastened petal abscission in Torenia (Goto et al., 1999). In Delphinium,
wounding of ovaries or receptacles accelerates abscission of flowers
(Ichimura et al., 2009a).
In wounding-induced flower senescence, ethylene production is has-
tened, as observed in pollination-induced flower senescence. However,
the pattern of ethylene synthesis induced by wounding differs from that
induced by pollination in petunia (Whitehead et al., 1984; Hoekstra and
Weges, 1996). In pollinated petunia flowers, ethylene is exclusively pro-
duced by the stigma and style region, whereas wounding of the stigma
induces ethylene production both by the stigma and style region and by the
remaining flower parts (Woltering et al., 1997). These observations sug-
gest that the mechanisms that accelerate petal senescence are different in
pollination- and wound-induced petal senescence.

4.2.9  Effect of Temperature on Ethylene

Production and Perception
Ethylene production and perception are affected by temperature. In carna-
tion, the time to induction of petal senescence by exogenous ethylene is
prolonged under lower temperatures, suggesting that ethylene sensitivity
declines as temperature falls (Barden and Hanan, 1972). However, it is not
clear whether this decline in ethylene sensitivity is a unique phenomenon
among other physiological responses to low temperature, such as a decline
in respiration. In contrast, lily flowers, which have relatively low ethylene
sensitivity, show an increase in ethylene sensitivity when flowers are kept
at low temperature (Song and Peng, 2004).
Ethylene synthesis is suppressed by high temperature in fruits such
as tomato and apple (Lurie, 1998). In carnation flowers, high temperature
suppresses ethylene synthesis (Yangkhamman et al., 2005, 2007; Shibuya
and Ichimura, 2010). When cut flowers of carnation ‘Barbara’ are kept at

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more than 32°C, ethylene production from the flowers is suppressed,

­resulting in a delay in petal wilting. However, flowers kept at such a high
temperature show inhibited petal growth and fading of color.

4.3  Plant Hormones Other than

Ethylene in Flower Senescence
Plant hormones include auxin, gibberellin, cytokinin, abscisic acid (ABA),
brassinolides, and jasmonic acid. Salicylic acid and strigolactones are also
considered plant hormones. Of these, auxin, gibberellin, cytokinin, abscisic
acid, and jasmonic acid have been analyzed in studies on flower senescence.

4.3.1 Auxin
Auxin at relatively high concentrations induces ethylene synthesis. In
carnation, treatment with indoleacetic acid or 2,4-dichlorophenoxyacetic
acid, a synthetic auxin, induces ethylene production, leading to hastened
petal senescence (Sacalis and Nichols, 1980; Wulster et al., 1982). Auxin
at high concentrations seems to induce ACC synthase activity, resulting in
an increase in ethylene production. In contrast, application of 1-naphtha­
leneacetic acid, another synthetic auxin, suppresses abscission of bougain-
villea flowers (Chang and Chen, 2001).

4.3.2 Gibberellin
Gibberellin treatment prolongs flower longevity by suppressing ethylene
synthesis in carnation (Saks et al., 1992). It also prolongs the vase life of
daffodil, which has relatively low ethylene sensitivity (Ichimura and Goto,
2002). Since it is difficult to accurately quantify the gibberellin content in
plants, little is known of the distribution and movement of gibberellins in
cut flowers.

4.3.3 Cytokinin
Cytokinin generally delays senescence in plants. In rose (Mayak and
Halevy, 1970) and carnation (van Staden et al., 1987), cytokinin-like activity
decreases in senescent flowers. Application of benzyladenine, a synthetic
cytokinin, to presenescent cut carnation flowers delays petal senescence
(Mayak and Dilley, 1976a; Eisinger, 1977; Cook et al., 1985).

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Cytokinin treatment prevents the conversion of exogenously applied

ACC to ethylene in carnation petals, and ethylene treatment does not
induce endogenous ethylene synthesis in the petals of flowers treated with
­c ytokinin (Mor et al., 1983; Cook et al., 1985). However, cytokinin treatment
does not inhibit ethylene synthesis in flowers in which a rise in ­ethylene syn-
thesis has already started (Mor et al., 1983). These observations suggest
that cytokinin does not directly inhibit the activities of enzymes involved
in e­ thylene synthesis, but it may suppress the synthesis of enzymes or
decrease ethylene sensitivity.
A synthetic cytokinin, thidiazuron, has been reported to improve
the vase life of iris flowers, which have relatively low ethylene sensitivity
(Macnish et al., 2010). In transgenic petunia expressing bacterial ipt, which
encodes a key enzyme of cytokinin synthesis, petal senescence is delayed
as a result of an increase in cytokinin content (Chang et al., 2003). In these
transgenic plants, the peak of ethylene production from flowers is delayed,
ethylene sensitivity declines, and ABA content decreases.
Cytokinin is likely involved in the regulation of flower senescence
in many plant species, regardless of the degree of ethylene sensitivity.
However, it is largely unknown whether changes in cytokinin content occur
in senescing tissues.

4.3.4  Abscisic Acid

ABA is generally considered to be a plant hormone that accelerates
senescence. ABA content in petals increases as the petals senesce in
rose (Borochov et al., 1976b) and carnation (Eze et al., 1986; Hanley and
Bramlage, 1989). In carnation, ABA treatment accelerates ethylene produc-
tion and increases ethylene sensitivity (Mayak and Dilley, 1976a, 1976b).
On the other hand, ethylene treatment increases ABA content in carnation.
In flowers with low ethylene sensitivity, such as daylily (Panavas
et al., 1998) and daffodil (Hunter et al., 2004a), ABA treatment also hastens
flower senescence. In daffodil, ABA concentration in the perianth increases
as flowers senesce (Hunter et al., 2004a).
ABA treatment of cut roses with leaves removed accelerates petal
senescence, while it delays senescence when the leaves are not removed
(Halevy et al., 1974; Borochov et al., 1976a). This observation is explained
as follows: when the leaves are removed, ABA directly affects the petals
and hastens petal senescence; in cut roses with leaves, ABA suppresses
transpiration from the leaves, resulting in improved water uptake, leading
to the prolongation of vase life. In Eustoma, ABA treatment extends vase
life by improving the water conditions of cut flowers (Shimizu-Yumoto and
Ichimura, 2009a).

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ABA generally accelerates flower senescence, but it sometimes

improves the vase life of cut flowers by suppressing transpiration, leading
to better water conditions.

4.3.5  Jasmonic Acid

There are few reports on the effect of jasmonic acid on flower senescence.
Treatment with jasmonic acid ethyl ester accelerates flower senescence in
petunia (Porat and Halevy, 1993), Phalaenopsis, and Dendrobium (Porat
et al., 1995b). Since inhibitors of jasmonic acid synthesis have been devel-
oped, these inhibitors can be used to delay flower senescence.

4.4  Programmed Cell Death in Flower Senescence

4.4.1  Programmed Cell Death

Flower longevity is species specific and is thought to be genetically con-
trolled. Treatment with cycloheximide, a protein synthesis inhibitor, delays
flower senescence in many plant species, including carnation (Ichimura
et al., 2009b), gladiolus (Jones et al., 1994; Yamane and Ogata, 1995), and
Japanese morning glory (Yamada et al., 2009). The result of cycloheximide
treatment in gladiolus is shown in Figure 4.2. In Japanese morning glory,
actinomycin D, which inhibits RNA synthesis, also delays petal senes-
cence (Yamada et al., 2009). Furthermore, suppression of expression of the

Figure 4.2  Effect of cycloheximide, a protein synthesis inhibitor, on flower

senescence of gladiolus flowers. Detached flowers were placed in distilled
water (left) or 1 mM cycloheximide for 4 days.

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gene for deoxyhypusine hydroxylase, which is involved in the synthesis of

eukaryotic translation initiation factor 5A (eIF5A), results in delayed petal
senescence in carnation (Hopkins et al., 2007). Based on these observa-
tions, flower senescence is considered to be a type of programmed cell
death (PCD), which is an actively regulated process.
A typical hallmark of PCD in animal cells is the degradation of nuclei
and DNA. During petal senescence, the first sign of PCD is chromatin
condensation. In petal cells of petal-wilting-type flowers, including petunia
(Xu and Hanson, 2000; Langston et al., 2005) and Japanese morning glory
(Yamada et al., 2006), DNA degradation is observed during s­ enescence.
In  contrast, DNA degradation is not observed in petal cells of Prunus
yedoensis, a type of cherry tree, which is a petal-abscission-type flower, at
the point of petal abscission (Yamada et al., 2007b), suggesting that PCD
may not occur in this flower until the petals abscise.

4.4.2  Gene Expression during PCD in Flowers

Changes in gene expression during flower senescence have been analyzed
by microarrays or subtraction assays in several species, including Iris
(Van Doorn et al., 2003), Japanese morning glory (Yamada et al., 2007a),
Alstroemeria (Breeze et al., 2004), Mirabilis jalapa (Xu et al., 2007), carnation
(Hoeberichts et al., 2007), daffodil (Hunter et al., 2002), and daylily (Panavas
et al., 1999). In petunia, proteomic analysis has also been conducted and pro-
tein changes have been profiled during petal senescence (Bai et al., 2010).
Genes that are commonly upregulated during flower senescence in these
plant species are genes encoding enzymes that are involved in the degrada-
tion of proteins or phospholipids, such as cysteine proteinase and aspartic
proteinase. In addition to these genes, the expression of genes for glutathione
transferase in carnation (Meyer et al., 1991) and in-chain fatty acid hydroxy-
lase, fatty acid elongase, and allene oxide synthase in daylily (Panavas et al.,
1999) has been reported to increase during senescence.
In PCD of animal cells, caspase, a cysteine proteinase, plays a
very important role. Caspase has not been isolated in plants, but vacu-
olar processing enzyme (VPE) is a protease that exhibits caspase a­ ctivity
(­Hara-Nishimura et al., 2005). VPE is localized in the vacuoles and is
responsible for the maturation or activation of vacuolar proteins. Expression
of VPE homologs increases during petal senescence in Japanese morn-
ing glory (Yamada et al., 2009), carnation (Hoeberichts et al., 2007), and
­daffodil (Hunter et al., 2002).
The Bax (Bcl-2-associated X protein) inhibitor inhibits Bax activity
in the PCD of animal cells. Bax proteins have not been found in plants, but
genes for Bax inhibitor proteins have been isolated in several plant species.
In Japanese morning glory, the mRNA level of Bax inhibitor increases as

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the petals senesce (Yamada et al., 2009). DAD1 (defender against apoptotic
cell death) is a protein that suppresses the progression of PCD in animal
cells. The mRNA level of a DAD1 homolog starts to decrease at early stages
of petal senescence in Alstroemeria (Breeze et al., 2004), while it decreases
at later stages in carnation (Hoeberichts et al., 2007), gladiolus (Yamada
et al., 2004), and Iris (Van Doorn et al., 2003). However, the role of DAD1 in
flower senescence is unknown.

4.4.3  Autophagy in Petal Senescence

Autophagy is often observed in plant cells at later stages of senescence.
Autophagy, or “self-eating,” is a conserved mechanism for the degradation
of cellular contents in eukaryotic species. This occurs by the uptake of cyto-
plasmic constituents into the vacuole, where they are degraded by vacuolar
hydrolases in plants (Bassham, 2007; Li and Vierstra, 2012). In senesc-
ing petals of morning glory (Ipomoea purpurea), numerous vesicles and
cytoplasmic components have been observed in the vacuoles by electron
microscopy, indicating autophagic processes (Matile and Winkenbach,
1971). Similar autophagic phenomena have been observed in senescing pet-
als of carnation (Smith et al., 1992).
Several genes involved in autophagic processes (autophagy-related
genes [ATGs]) have been identified in budding yeast (Saccharomyces
­cerevisiae) (Ohsumi, 2001), and ATG homologs have been isolated in
Arabidopsis thaliana (Bassham et al., 2006). mRNA levels for ATG4 and
ATG8 homologs in Japanese morning glory (Yamada et al., 2009; Shibuya
et al., 2009, 2011) and for ATG8 homologs in petunia increase during petal
senescence (Shibuya et al., 2013). The increase in mRNA levels of ATG
homologs may reflect the activation of autophagy; however, it is difficult to
accurately evaluate autophagic activity. Autophagy is proposed to play an
important role in nutrient recycling in leaf senescence (Bassham, 2007),
and this is likely the case for its role in petal senescence. Autophagy has
been thought to be closely associated with cell death in plants; however, it
is unclear whether autophagy is the direct cause of cell death during petal
senescence.

4.5  Conclusions and Future Perspectives

Ethylene is clearly a key factor that regulates flower senescence in many
plant species. As ethylene biosynthetic and signaling pathways have been
well characterized, it is now possible to control the effects of ethylene by
chemical treatment or genetic modification. However, several issues in

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flower senescence remain to be elucidated. In particular, the mechanisms

regulating ethylene-independent senescence are largely unknown.
It is unclear how differences in responses to ethylene among plant spe-
cies arise. In flowers that exhibit very low ethylene sensitivity, such as chrysan-
themum and gladiola, genes for ethylene receptors are expressed in the petals
(Arora et al., 2006; Narumi et al., 2005). Thus, ethylene should be perceived
and the signal transmitted in these flowers as in flowers with high ethylene
sensitivity. The ethylene signal somehow may not switch on downstream path-
ways that induce cell death in flowers that do not respond to ethylene.
Important roles of the gynoecia or stamen in the regulation of petal
senescence are highlighted in flowers with high ethylene sensitivity, such
as carnation and Portulaca (Ichimura and Suto, 1998; Shibuya et al., 2000).
However, the detailed mechanisms of interorgan regulation in natural
senescence of flowers remain to be elucidated.
In flowers that show ethylene-independent senescence, little is known
of the regulatory pathways of petal senescence. Since inhibitors of RNA and
protein synthesis delay senescence in these flowers, genes that actively reg-
ulate senescence should exist. Although many genes that show changes in
expression during senescence have been isolated, key regulators of flower
senescence have not been identified. A lack of good model plants that are
transformable makes it difficult to determine the function of isolated genes.
Recently, Japanese morning glory has been proposed as a model plant to
study ethylene-independent senescence; it is already used for functional
analysis (Shibuya, 2012).
Physiological and molecular biological studies on flower senescence
have made substantial contributions to the development of postharvest
technologies in cut flowers. However, the mechanisms behind flower senes-
cence are not fully understood. Further advances in the knowledge of
postharvest biology will help the development of innovative technology to
improve the postharvest quality of flowers.

References
Alonso, J.M., Hirayama, T., Roman, G., Nourizadeh, S., and Ecker, J.R. 1999.
EIN2, a bifunctional transducer of ethylene and stress responses in
Arabidopsis. Science 284, 2148–2152.
Arora, A., Watanabe, S., Ma, B., Takada, K., and Ezura, H. 2006. A novel eth-
ylene receptor homolog gene isolated from ethylene-insensitive flowers
of gladiolus (Gladiolus grandiflora Hort.). Biochemical and Biophysical
Research Communications 351, 739–744.
Bai, S.Y., Willard, B., Chapin, L.J., Kinter, M.T., Francis, D.M., Stead, A.D.,
and Jones, M.L. 2010. Proteomic analysis of pollination-induced corolla
senescence in petunia. Journal of Experimental Botany 61, 1089–1109.

125
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Barden, L.E., and Hanan, J.J. 1972. Effect of ethylene on carnation keeping
life. Journal of the American Society for Horticultural Science 97, 785–788.
Bassham, D.C. 2007. Plant autophagy—more than a starvation response.
Current Opinion in Plant Biology 10, 587–593.
Bassham, D.C., Laporte, M., Marty, F., Moriyasu, Y., Ohsumi, Y., Olsen,
L.J., and Yoshimoto, K. 2006. Autophagy in development and stress
responses of plants. Autophagy 2, 2–11.
Borochov, A., Mayak, S., and Halevy, A.H. 1976a. Combined effects of abscisic
acid and sucrose on growth and senescence of rose flowers. Physiologia
Plantarum 36, 221–224.
Borochov, A., Tirosh, T., and Halevy, A.H. 1976b. Abscisic acid content of
senescing petals on cut rose flowers as affected by sucrose and water
stress. Plant Physiology 58, 175–178.
Breeze, E., Wagstaff, C., Harrison, E., Bramke, I., Rogers, H., Stead, A.,
Thomas, B., and Buchanan-Wollaston, V. 2004. Gene expression pat-
terns to define stages of postharvest senescence in Alstroemeria petals.
Plant Biotechnology Journal 2, 155–168.
Burg, S.P., and Dijkman, M.J. 1967. Ethylene and auxin participation in
pollen induded fading of Vanda orchid blossoms. Plant Physiology 42,
1648–1650.
Celikel, F.G., and Reid, M.S. 2002. Postharvest handling of stock (Matthiola
incana). HortScience 37, 144–147.
Chang, C., Kwok, S.F., Bleecker, A.B., and Meyerowitz, E.M. 1993. Arabidopsis
ethylene-response gene ETR1: Similarity of product to two-component
regulators. Science 262, 539–544.
Chang, H., Jones, M.L., Banowetz, G.M., and Clark, D.G. 2003. Overproduction
of cytokinins in petunia flowers transformed with PSAG12-IPT delays
corolla senescence and decreases sensitivity to ethylene. Plant
Physiology 132, 2174–2183.
Chang, Y.S., and Chen, H.C. 2001. Variability between silver thiosulfate and
1-naphthaleneacetic acid applications in prolonging bract longevity of
potted bougainvillea. Scientia Horticulturae 87, 217–224.
Chao, Q., Rothenberg, M., Solano, R., Roman, G., Terzaghi, W., and Ecker, J.R.
1997. Activation of the ethylene gas response pathway in Arabidopsis by
the nuclear protein ETHYLENE-INSENSITIVE3 and related proteins.
Cell 89, 1133–1144.
Clark, D.G., Richards, C., Hilioti, Z., LindIversen, S., and Brown, K. 1997.
Effect of pollination on accumulation of ACC synthase and ACC oxidase
transcripts, ethylene production and flower petal abscission in gera-
nium (Pelargonium × hortorum L.H. Bailey). Plant Molecular Biology
34, 855–865.
Cook, D., Rasche, M., and Eisinger, W. 1985. Regulation of ethylene bio-
synthesis and action in cut carnation flower senescence by cytoki-
nins. Journal of the American Society for Horticultural Science 110,
24–27.

126
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Dervinis, C., Clark, D.G., Barrett, J.E., and Nell, T.A. 2000. Effect of polli-
nation and exogenous ethylene on accumulation of ETR1 homologue
transcripts during flower petal abscission in geranium (Pelargonium ×
hortorum L.H. Bailey). Plant Molecular Biology 42, 847–856.
Doi, M., Nakagawa, Y., Watabe, S., Aoe, K., Inamoto, K., and Imanishi,  H.
2003. Ethylene-induced leaf yellowing in cut chrysanthemums
(Dendranthema grandiflora Kitamura). Journal of the Japanese Society
for Horticultural Science 72, 533–535.
Eisinger, W. 1977. Role of cytokinins in carnation flower senescence. Plant
Physiology 59, 707–709.
Elgar, H.J., Woolf, A.B., and Bieleski, R.L. 1999. Ethylene production by three
lily species and their response to ethylene exposure. Postharvest Biology
and Technology 16, 257–267.
Eze, J.M.O., Mayak, S., Thompson, J.E., and Dumbroff, E.B. 1986. Senescence
in cut carnation flowers: temporal and physiological relationships
among water status, ethylene, abscisic acid and membrane permeability.
Physiologia Plantarum 68, 323–328.
Fromm, J., Hajirezaei, M., and Wilke, I. 1995. The biochemical response of
electrical signaling in the reproductive system of Hibiscus plants. Plant
Physiology 109, 375–384.
Gilissen, L.J.W. 1977. Style-controlled wilting of flower. Planta 133, 275–280.
Gilissen, L.J.W., and Hoekstra, F.A. 1984. Pollination-induced corolla wilting
in Petunia hybrida rapid transfer through the style of a wilting-inducing
substance. Plant Physiology 75, 496–498.
Goh, C.J., Halevy, A.H., Engel, R., and Kofranek, A.M. 1985. Ethylene evolution
and sensitivity in cut orchid flowers. Scientia Horticulturae 26, 57–67.
Goto, R., Aida, R., Shibata, M., and Ichimura, K. 1999. Role of ethylene
on flower senescence of Torenia. Journal of the Japanese Society for
Horticultural Science 68, 263–268.
Guo, H., and Ecker, J.R. 2003. Plant responses to ethylene gas are mediated
by SCF(EBF1/EBF2)-dependent proteolysis of EIN3 transcription
­factor. Cell 115, 667–677.
Halevy, A.H., Mayak, S., Tirosh, T., Spiegelstein, H., and Kofranek, A.M.
1974. Opposing effects of abscisic acid on senescence of rose flowers.
Plant and Cell Physiology 15, 813–821.
Hanley, K.M., and Bramlage, W.J. 1989. Endogenous levels of abscisic acid
in aging carnation flower parts. Journal of Plant Growth Regulation 8,
225–236.
Hara-Nishimura, I., Hatsugai, N., Nakaune, S., Kuroyanagi, M., and
Nishimura, M. 2005. Vacuolar processing enzyme: An executor of plant
cell death. Current Opinion in Plant Biology 8, 404–408.
Henskens, J.A., Rouwendal, G.J., ten Have, A., and Woltering, E.J. 1994.
Molecular cloning of two different ACC synthase PCR fragments in
carnation flowers and organ-specific expression of the corresponding
genes. Plant Molecular Biology 26, 453–458.

127
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Hiraya, T., Shimizu, H., and Ichimura, K. 2002. Effects of STS, 1-MCP
and sucrose on the vase life of cut Oxypetalum caeruleum flowers.
Horticulture Research (Japan), 1, 67–70 (in Japanese with English
abstract).
Hoeberichts, F.A., Van Doorn, W.G., Vorst, O., Hall, R.D., and van Wordragen,
M.F. 2007. Sucrose prevents up-regulation of senescence-associated
genes in carnation petals. Journal of Experimental Botany 58, 2873–2885.
Hoekstra, F.A., and Weges, R. 1986. Lack of control by early pistillate ethylene
of the accelerated wilting of Petunia hybrida flowers. Plant Physiology 80,
403–408.
Hopkins, M., Taylor, C., Liu, Z., Ma, F., McNamara, L., Wang, T.W., and
Thompson, J.E. 2007. Regulation and execution of molecular disassem-
bly and catabolism during senescence. New Phytology 175, 201–214.
Hua, J., and Meyerowitz, E.M. 1998. Ethylene responses are negatively regu-
lated by a receptor gene family in Arabidopsis thaliana. Cell 94, 261–271.
Hua, J., Chang, C., Sun, Q., and Meyerowitz, E.M. 1995. Ethylene insensitiv-
ity conferred by Arabidopsis ERS gene. Science 269, 1712–1714.
Huang, Y., Li, H., Hutchison, C.E., Laskey, J., and Kieber, J.J. 2003. Biochemical
and functional analysis of CTR1, a protein kinase that negatively regu-
lates ethylene signaling in Arabidopsis. Plant Journal 33, 221–233.
Hunter, D.A., Ferrante, A., Vernieri, P., and Reid, M.S. 2004a. Role of abscisic
acid in perianth senescence of daffodil (Narcissus pseudonarcissus
‘Dutch Master’). Physiologia Plantarum 121, 313–321.
Hunter, D.A., Steele, B.C., and Reid, M.S. 2002. Identification of genes associ-
ated with perianth senescence in daffodil (Narcissus pseudonarcissus L.
‘Dutch Master’). Plant Science 163, 13–21.
Hunter, D.A., Yi, M.F., Xu, X.J., and Reid, M.S. 2004b. Role of ethylene in
perianth senescence of daffodil (Narcissus pseudonarcissus L. ‘Dutch
Master’). Postharvest Biology and Technology 32, 269–280.
Ichimura, K. 2010. Postharvest physiology of cut flowers: progress and future
aspects. Bulletin of National Institute of Floriculture Sciences 10, 11–53
(in Japanese).
Ichimura, K., and Goto, R. 2000. Acceleration of senescence by pollination of
cut ‘Asuka-no-nami’ Eustoma flowers. Journal of the Japanese Society for
Horticultural Science 69, 166–170.
Ichimura, K., and Goto, R. 2002. Extension of vase life of cut Narcissus tazetta
var. chinensis flowers by combined treatment with STS and gibberellin A3.
Journal of the Japanese Society for Horticultural Science 71, 226–230.
Ichimura, K., and Suto, K. 1998. Role of ethylene in acceleration of flower
senescence by filament wounding in Portulaca hybrid. Physiologia
Plantarum 104, 603–607.
Ichimura, K., Kishimoto, M., Norikoshi, R., Kawabata, Y., and Yamada, K.
2005. Soluble carbohydrates and variation in vase-life of cut rose cultivars
‘Delilah’ and ‘Sonia’. Journal of Horticultural Science and Biotechnology
80, 280–286.

128
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Ichimura, K., Niki, T., and Shimizu-Yumoto, H. 2007. Factors involved in the
vase life of cut snapdragon flowers. Horticulture Research (Japan) 6
(Suppl. 2), 341 (in Japanese).
Ichimura, K., Shimamura, M., and Hisamatsu, T. 1998. Role of ethylene in
senescence of cut Eustoma flowers. Postharvest Biology and Technology
14, 193–198.
Ichimura, K., Shimizu-Yumoto, H., and Goto, R. 2009a. Ethylene produc-
tion by gynoecium and receptacle is associated with sepal abscission
in cut Delphinium flowers. Postharvest Biology and Technology 52,
267–272.
Ichimura, K., Yamada, T., Yoshida, S., Pun, U.K., Tanase, K., and Shimizu-
Yumoto, H. 2009b. Ethylene regulates programmed cell death
(PCD) associated with petal senescence in carnation flowers. Acta
Horticulturae 847, 185–190.
Iordachescu, M., and Verlinden, S. 2005. Transcriptional regulation of three
EIN3-like genes of carnation (Dianthus caryophyllus L. cv. Improved
White Sim) during flower development and upon wounding, polli-
nation,  and ethylene exposure. Journal of Experimental Botany 56,
2011–2018.
Jones, M.L., and Woodson, W.R. 1997. Pollination-induced ethylene in carna-
tion: role of stylar ethylene in corolla senescence. Plant Physiology 115,
205–212.
Jones, M.L., and Woodson, W.R. 1999a. Differential expression of three mem-
bers of the 1-aminocyclopropane-1-carboxylate synthase gene family in
carnation. Plant Physiology 119, 755–764.
Jones, M.L., and Woodson, W.R. 1999b. Inter-organ signaling following pol-
lination in carnations. Journal of the American Society for Horticultural
Science 124, 598–604.
Jones, R.B., Serek, M., Kuo, C.L., and Reid, M.S. 1994. The effect of protein
synthesis inhibition on petal senescence in cut bulb flowers. Journal of
the American Society for Horticultural Science 119, 1243–1247.
Kato, M., Shimizu, H., Onozaki, T., Tanikawa, N., Ikeda, H., Hisamatsu, T.,
and Ichimura, K. 2002. Role of ethylene in senescence of pollinated
and unpollinated Campanula medium flowers. Journal of the Japanese
Society for Horticultural Science 71, 385–387.
Kende, H. 1993. Ethylene biosynthesis. Annual Review of Plant Physiology
and Plant Molecular Biology 44, 283–307.
Kevany, B.M., Tieman, D.M., Taylor, M.G., Cin, V.D., and Klee, H.J. 2007.
Ethylene receptor degradation controls the timing of ripening in tomato
fruit. Plant Journal 51, 458–467.
Kieber, J.J., Rothenberg, M., Roman, G., Feldmann, K.A., and Ecker, J.R.
1993. CTR1, a negative regulator of the ethylene response pathway in
Arabidopsis, encodes a member of the raf family of protein kinases. Cell
72, 427–441.

129
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Kosugi, Y., Shibuya, K., Tsuruno, N., Iwazaki, Y., Mochizuki, A., Yoshioka,
T., Hashiba, T., and Satoh, S. 2000. Expression of genes responsible for
ethylene production and wilting are differently regulated in carnation
(Dianthus caryophyllus L.) petals. Plant Science 158, 139–145.
Kuroda, S., Hakata, M., Hirose, Y., Shiraishi, M., and Abe, S. 2003. Ethylene
production and enhanced transcription of an ethylene receptor gene,
ERS1, in Delphinium during abscission of florets. Plant Physiology and
Biochemistry 41, 812–820.
Kuroda, S., Hirose, Y., Shiraishi, M., Davies, E., and Abe, S. 2004. Co-expression
of an ethylene receptor gene, ERS1, and ethylene signaling regulator
gene, CTR1, in Delphinium during abscission of florets. Plant Physiology
and Biochemistry 42, 745–751.
Langston, B.J., Bai, S., and Jones, M.L. 2005. Increases in DNA fragmenta-
tion and induction of a senescence-specific nuclease are delayed during
corolla senescence in ethylene-insensitive (etr1-1) transgenic petunias.
Journal of Experimental Botany 56, 15–23.
Larsen, P.B., Ashworth, E.N., Jones, M.L., and Woodson, W.R. 1995.
Pollination-induced ethylene in carnation: Role of pollen tube growth
and sexual compatibility. Plant Physiology 108, 1405–1412.
Li, F., and Vierstra, R.D. 2012. Autophagy: a multifaceted intracellular system
for bulk and selective recycling. Trends in Plant Science 17, 526–537.
Lin, Z., Zhong, S., and Grierson, D. 2009. Recent advances in ethylene
research. Journal of Experimental Botany 60, 3311–3336.
Lindstrom, J.T., Lei, C.H., Jones, M.L., and Woodson, W.R. 1999. Accumulation
of 1-aminocyclopropane-1-carboxylic acid (ACC) in petunia pollen is
associated with expression of a pollen-specific ACC synthase late in
development. Journal of the American Society for Horticultural Science
124, 145–151.
Lovell, P.J., Lovell, P.H., and Nichols, R. 1987. The control of flower senes-
cence in petunia (Petunia hybrida). Annals of Botany 60, 49–59.
Lurie, S. 1998. Postharvest heat treatments. Postharvest Biology and
Technology 14, 257–269.
Macnish, A.J., Jiang, C.Z., and Reid, M.S. 2010. Treatment with thidiazuron
improves opening and vase life of iris flowers. Postharvest Biology and
Technology 56, 77–84.
Matile, P., and Winkenbach, F. 1971. Function of lysosomes and lysosomal
enzymes in the senescing corolla of the morning glory (Ipomoea
­purpurea). Journal of Experimental Botany 22, 759–771.
Mayak, S., and Dilley, D.R. 1976a. Effect of sucrose on response of cut car-
nation to kinetin, ethylene, and abscisic acid. Journal of the American
Society for Horticultural Science 101, 583–585.
Mayak, S., and Dilley, D.R. 1976b. Regulation of senescence in carnation
(Dianthus caryophyllus): Effect of abscisic acid and carbon dioxide on
ethylene production. Plant Physiology 58, 663–665.
Mayak, S., and Halevy, A.H. 1970. Cytokinin activity in rose petals and its rela-
tion to senescence. Plant Physiology 46, 497–499.

130
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Meyer Jr., R.C., Goldsbrough, P.B., and Woodson, W.R. 1991. An ethylene-
responsive flower senescence-related gene from carnation encodes
a protein homologous to glutathione s-transferases. Plant Molecular
Biology 17, 277–281.
Momonoi, K., Shoji, K., and Yoshida, K. 2007. Cloning and characterization of
ACC oxidase genes from tulip. Plant Biotechnology 24, 241–246.
Mor, Y., Halevy, A.H., Spiegelstein, H., and Mayak, S. 1985. The site of
1-­aminocyclopropane-1-carboxylic acid synthesis in senescing carna-
tion ­petals. Physiologia Plantarum 65, 196–202.
Mor, Y., Reid, M.S., and Kofranek, A.M. 1980. Role of the ovary in carnation
senescence. Scientia Horticulturae 13, 377–383.
Mor, Y., Reid, M.S., and Kofranek, A.M. 1984. Pulse treatments with silver
thiosulfate and sucrose improve the vase life of sweet peas. Journal of
the American Society for Horticultural Science 109, 866–868.
Mor, Y., Spiegelstein, H., and Halevy, A.H. 1983. Inhibition of ethylene
biosynthesis in carnation petals by cytokinin. Plant Physiology 71,
541–546.
Müller, R., Lind-Iversen, S., Stummann, B.M., and Serek, M. 2000a. Expression
of genes for ethylene biosynthetic enzymes and an ethylene receptor
in senescing flowers of miniature potted roses. Journal of Horticultural
Science and Biotechnology 75, 12–18.
Müller, R., Owen, C.A., Xue, Z.T., Welander, M., and Stummann, B.M. 2002.
Characterization of two CTR-like protein kinases in Rosa hybrida and
their expression during flower senescence and in response to ethylene.
Journal of Experimental Botany 53, 1223–1225.
Müller, R., Owen, C.A., Xue, Z.T., Welander, M., and Stummann, B.M. 2003.
The transcription factor EIN3 is constitutively expressed in miniature
roses with differences in postharvest life. Journal of horticultural sci-
ence & biotechnology 78, 10–14.
Müller, R., Stummann, B.M., and Serek, M. 2000b. Characterization of an
ethylene receptor family with differential expression in rose (Rosa
­hybrida L.) flowers. Plant Cell Reports 19, 1232–1239.
Nadeau, J.A., Xian, S.Z., Nair, H., and O’neill, S.D. 1993. Temporal and spa-
tial regulation of 1-aminocyclopropane-1-carboxylate oxidase in the
p­ollination-induced senescence of orchid flowers. Plant Physiology 103,
31–39.
Narumi, T., Kanno, Y., Suzuki, M., Kishimoto, S., Ohmiya, A., and Satoh, S.
2005. Cloning of a cDNA encoding an ethylene receptor (DG-ERS1)
from chrysanthemum and comparison of its mRNA level in ethylene-
sensitive and -insensitive cultivars. Postharvest Biology and Technology
36, 21–30.
Nichols, R. 1966. Ethylene production during senescence of flowers. Journal
of Horticultural Science and Biotechnology 41, 279–290.
Nichols, R. 1968. The response of carnations (Dianthus caryophyllus) to eth-
ylene. Journal of Horticultural Science 43, 335–349.

131
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Norikoshi, R., Niki, T., and Ichimura, K. 2008. Changes in ACO activity and
two ACO gene expression in petals, styles and ovary during flower
senescence in cut carnations. Horticulture Research (Japan) 7 (Suppl. 2),
330 (in Japanese).
Ohsumi, Y. 2001. Molecular dissection of autophagy: two ubiquitin-like sys-
tems. National Review of Molecular Cell Biology 2, 211–216.
Okamoto, M., and Ichimura, K. 2010. Changes in ethylene production by pol-
lination in cut Delphinium grandiflorum. Horticulture Research (Japan)
9 (Suppl. 1), 218 (in Japanese).
O’Neill, S.D., Nadeau, J.A., Zhang, X.S., Bui, A.Q., and Halevy, A.H. 1993.
Interorgan regulation of ethylene biosynthetic genes by pollination.
Plant Cell 5, 419–432.
Onozaki, T., Ikeda, H., and Shibata, M. 2004. Video evaluation of ethylene
sensitivity after anthesis in carnation (Dianthus caryophyllus L.) flowers.
Scientia Horticulturae 99, 187–197.
Panavas, T., Pikula, A., Reid, P.D., Rubinstein, B., and Walker, E.L. 1999.
Identification of senescence-associated genes from daylily petals. Plant
Molecular Biology 40, 237–248.
Panavas, T., Walker, E.L., and Rubinstein, B. 1998. Possible involvement of
abscisic acid in senescence of daylily petals. Journal of Experimental
Botany 49, 1987–1997.
Park, K.Y., Drory, A., and Woodson, W.R. 1992. Molecular cloning of an
1-aminocyclopropane-1-carboxylate synthase from senescing carnation
flower petals. Plant Molecular Biology 18, 377–386.
Porat, R., and Halevy, A.H. 1993. Enhancement of petunia and dendrobium
flower senescence by jasmonic acid methyl ester is via the promotion of
ethylene production. Plant Growth Regulation 13, 297–301.
Porat, R., Borochov, A., and Halevy, A.H. 1994. Pollination-induced changes
in ethylene production and sensitivity to ethylene in cut dendrobium
orchid flowers. Scientia Horticulturae 58, 215–221.
Porat, R., Halevy, A.H., Serek, M., and Borochov, A. 1995a. An increase in
ethylene sensitivity following pollination is the initial event trigger-
ing an increase in ethylene production and enhanced senescence of
Phalaenopsis orchid flowers. Physiologia Plantarum 93, 778–784.
Porat, R., Reiss, N., Atzorn, R., Halevy, A.H., and Borochov, A. 1995b.
Examination of the possible involvement of lipoxygenase and jasmo-
nates in pollination-induced senescence of Phalaenopsis and Dendrobium
orchid flowers. Physiologia Plantarum 94, 205–210.
Qiao, H., Chang, K.N., Yazaki, J., and Ecker, J.R. 2009. Interplay between
­ethylene, ETP1/ETP2 F-box proteins, and degradation of EIN2 triggers
ethylene responses in Arabidopsis. Genes and Development 23, 512–521.
Qiao, H., Shen, Z.X., Huang, S.S.C., Schmitz, R.J., Urich, M.A., Briggs,
S.P., and Ecker, J.R. 2012. Processing and subcellular trafficking of
ER-tethered EIN2 control response to ethylene gas. Science 338,
390–393.

132
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Reid, M.S., Fujino, D.W., Hoffman, N.E., and Whitehead, C.S. 1984.
1-Aminocyclopropane-1-carboxylic acid (ACC): The transmitted
­stimulus  in pollinated flowers. Journal of Plant Growth Regulation 3,
189–196.
Rogers, H.J. 2006. Programmed cell death in floral organs: How and why do
flowers die? Annals of Botany 97, 309–315.
Rogers, H.J. 2013. From models to ornamentals: How is flower senescence
regulated? Plant Molecular Biology 82, 563–574.
Sacalis, J.N., and Lee, J.S. 1987. Promotion of floral longevity by the ovary
in carnation flowers. Journal of the American Society for Horticultural
Science 112, 118–121.
Sacalis, J.N., and Nichols, R. 1980. Effects of 2,4-D uptake on petal senes-
cence in cut carnation flowers. HortScience 15, 499–500.
Sakai, H., Hua, J., Chen, Q.G., Chang, C., Medrano, L.J., Bleecker, A.B., and
Meyerowitz, E.M. 1998. ETR2 is an ETR1-like gene involved in ethylene
signaling in Arabidopsis. Proceedings of the National Academy of Sciences
USA 95, 5812–5817.
Saks, Y., Vanstaden, J., and Smith, M.T. 1992. Effect of gibberellic acid on car-
nation flower senescence: Evidence that the delay of carnation flower
senescence by gibberellic acid depends on the stage of flower develop-
ment. Plant Growth Regulation 11, 45–51.
Serek, M., Jones, R.B., and Reid, M.S. 1994. Role of ethylene in opening and
senescence of Gladiolus sp. flowers. Journal of the American Society for
Horticultural Science 119, 1014–1019.
Sexton, R., Laird, G., and Van Doorn, W.G. 2000. Lack of ethylene involve-
ment in tulip petal abscission. Physiologia Plantarum 108, 321–329.
Shibuya, K. 2012. Molecular mechanisms of petal senescence in ornamental
plants. Journal of the Japanese Society for Horticultural Science 81, 140–149.
Shibuya, K., and Clark, D.G. 2006. Ethylene: Current status and future direc-
tions of using transgenic techniques to improve flower longevity of
ornamental crops. Journal of Crop Improvement 18, 391–412.
Shibuya, K., and Ichimura, K. 2010. Depression of autocatalytic ethylene pro-
duction by high-temperature treatment in carnation flowers. Journal of
the Japanese Society for Horticultural Science 79, 97–102.
Shibuya, K., Barry, K.G., Ciardi, J.A., Loucas, H.M., Underwood, B.A.,
Nourizadeh, S., Ecker, J.R., Klee, H.J., and Clark, D.G. 2004. The central
role of PhEIN2 in ethylene responses throughout plant development in
petunia. Plant Physiology 136, 2900–2912.
Shibuya, K., Nagata, M., Tanikawa, N., Yoshioka, T., Hashiba, T., and
Satoh, S. 2002. Comparison of mRNA levels of three ethylene receptors
in senescing flowers of carnation (Dianthus caryophyllus L.). Journal of
Experimental Botany 53, 399–406.
Shibuya, K., Niki, T., and Ichimura, K. 2013. Pollination induces a­ utophagy
in petunia petals via ethylene. Journal of Experimental Botany 64,
1111–1120.

133
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Shibuya, K., Shimizu, K., Yamada, T., and Ichimura, K. 2011. Expression of
autophagy-associated ATG8 genes during petal senescence in Japanese
morning glory. Journal of the Japanese Society for Horticultural Science
80, 89–95.
Shibuya, K., Yamada, T., Suzuki, T., Shimizu, K., and Ichimura, K. 2009.
InPSR26, a putative membrane protein, regulates programmed cell
death during petal senescence in Japanese morning glory. Plant
Physiology 149, 816–824.
Shibuya, K., Yoshioka, T., Hashiba, T., and Satoh, S. 2000. Role of the gynoe-
cium in natural senescence of carnation (Dianthus caryophyllus L.)
­flowers. Journal of Experimental Botany 51, 2067–2073.
Shimizu, H., and Ichimura, K. 2001. Involvement of ethylene in accelera-
tion of flower senescence by filament touching in Portulaca hybrid.
Horticulture Research (Japan) 70 (Suppl. 2), 467 (in Japanese).
Shimizu-Yumoto, H., and Ichimura, K. 2006a. Involvement of high sensitivity of
flag petal to ethylene on its closing in cut sweet pea flowers. Journal of the
Japanese Society for Horticultural Science 75 (Suppl. 1), 246 (in Japanese).
Shimizu-Yumoto, H., and Ichimura, K. 2006b. Senescence of Eustoma flowers
as affected by pollinated area of the stigmatic surface. Journal of the
Japanese Society for Horticultural Science 75, 66–71.
Shimizu-Yumoto, H., and Ichimura, K. 2009a. Abscisic acid, in combination
with sucrose, is effective as a pulse treatment to suppress leaf dam-
age and extend foliage vase-life in cut Eustoma flowers. Journal of
Horticultural Science and Biotechnology 84, 107–111.
Shimizu-Yumoto, H., and Ichimura, K. 2009b. Cultivar variation in the vase life
of unpollinated Eustoma flowers associated with ethylene. Horticulture
Research (Japan) 8, 359–364 (in Japanese with English abstract).
Singh, A., Evensen, K.B., and Kao, T.H. 1992. Ethylene synthesis and flo-
ral senescence following compatible and incompatible pollinations in
Petunia inflata. Plant Physiology 99, 38–45.
Smith, M.T., Saks, Y., and van Staden, J. 1992. Ultrastructural changes in the
petals of senescing flowers of Dianthus caryophyllus L. Annals of Botany
69, 277–285.
Solano, R., Stepanova, A., Chao, Q., and Ecker, J.R. 1998. Nuclear events in
ethylene signaling: a transcriptional cascade mediated by ETHYLENE-
INSENSITIVE3 and ETHYLENE-RESPONSE-FACTOR1. Genes and
Development 12, 3703–3714.
Song, L.L., and Peng, Y.H. 2004. Effect of cold storage on sensitivity of cut
lily to ethylene. Journal of Horticultural Science and Biotechnology 79,
723–728.
Spanjers, A.W. 1981. Bioelectric potential changes in the style of Lilium lon-
giflorum Thunb. after self- and cross-pollination of the stigma. Planta
153, 1–5.
Stead, A.D. 1992. Pollination-induced flower senescence—A review. Plant
Growth Regulation 11, 13–20.

134
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Stead, A.D., and Moore, K.G. 1979. Studies on flower longevity in Digitalis:
Pollination induced corolla abscission in Digitalis flowers. Planta 146,
409–414.
Tanase, K., and Ichimura, K. 2006. Expression of ethylene receptors
Dl-ERS1-3 and Dl-ERS2, and ethylene response during flower senes-
cence in Delphinium. Journal of Plant Physiology 163, 1159–1166.
Tang, X., Gomes, A., Bhatia, A., and Woodson, W.R. 1994. Pistil-specific and
ethylene-regulated expression of 1-aminocyclopropane-1-carboxylate
oxidase genes in petunia flowers. Plant Cell 6, 1227–1239.
Tang, X.Y., Wang, H., Brandt, A.S., and Woodson, W.R. 1993. Organization
and structure of the 1-aminocyclopropane-1-carboxylate oxidase gene
family from Petunia hybrida. Plant Molecular Biology 23, 1151–1164.
ten Have, A., and Woltering, E.J. 1997. Ethylene biosynthetic genes are
­differentially expressed during carnation (Dianthus caryophyllus L.)
flower senescence. Plant Molecular Biology 34, 89–97.
Tripathi, S.K., and Tuteja, N. 2007. Integrated signaling in flower senescence:
an overview. Plant Signaling and Behaviour 2, 437–445.
Van Doorn, W.G., and Woltering, E.J. 2008. Physiology and molecular biology
of petal senescence. Journal of Experimental Botany 59, 453–480.
Van Doorn, W.G., Balk, P.A., van Houwelingen, A.M., Hoeberichts, F.A., Hall,
R.D., Vorst, O., van der Schoot, C., and van Wordragen, M.F. 2003.
Gene expression during anthesis and senescence in Iris flowers. Plant
Molecular Biology 53, 845–863.
van Staden, J., Featonbysmith, B.C., Mayak, S., Spiegelstein, H., and Halevy,
A.H. 1987. Cytokinins in cut carnation flowers. II. Relationship between
endogenous ethylene and cytokinin levels in the petals. Plant Growth
Regulation 5, 75–86.
Wagstaff, C., Chanasut, U., Harren, F.J.M., Laarhoven, L.J., Thomas, B.,
Rogers, H.J., and Stead, A.D. 2005. Ethylene and flower longevity
in Alstroemeria: relationship between petal senescence, abscis-
sion and ethylene biosynthesis. Journal of Experimental Botany 56,
1007–1016.
Waki, K., Shibuya, K., Yoshioka, T., Hashiba, T., and Satoh, S. 2001. Cloning
of a cDNA encoding EIN3-like protein (DC-EIL1) and decrease in its
mRNA level during senescence in carnation flower tissues. Journal of
Experimental Botany 52, 377–379.
Wang, D., Fan, J., and Ranu, R.S. 2004. Cloning and expression of
1-­
aminocyclopropane-1-carboxylate synthase cDNA from rosa
(Rosa × ­hybrida). Plant Cell Reports 22, 422–429.
Wang, H., and Woodson, W.R. 1991. A flower senescence-related mRNA from
carnation shares sequence similarity with fruit ripening-related mRNAs
involved in ethylene biosynthesis. Plant Physiology 96, 1000–1001.
Wang, T.W., and Arteca, R.N. 1995. Identification and characterization of
cDNAs encoding ethylene biosynthetic enzymes from Pelargonium ×
hortorum cv Snow Mass leaves. Plant Physiology 109, 627–636.

135
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Wang, Y., and Kumar, P.P. 2007. Characterization of two ethylene receptors
PhERS1 and PhETR2 from petunia: PhETR2 regulates timing of anther
dehiscence. Journal of Experimental Botany 58, 533–544.
Whitehead, C.S., Halevy, A.H., and Reid, M.S. 1984. Roles of ethylene and
1-aminocyclopropane-1-carboxylic acid in pollination and woundin-
duced senescence of Petunia hybrida flowers. Physiologia Plantarum 61,
643–648.
Woltering, E.J., and Van Doorn, W.G. 1988. Role of ethylene in senescence
of petals—Morphological and taxonomical relationships. Journal of
Experimental Botany 39, 1605–1616.
Woltering, E.J., Balk, P.A., Nijenhuis-de Vries, M.A., Faivre, M., Ruys, G.,
Somhorst, D., Philosoph-Hadas, S., and Friedman, H. 2005. An auxin-
responsive 1-aminocyclopropane-1-carboxylate synthase is responsible
for differential ethylene production in gravistimulated Antirrhinum
majus L. flower stems. Planta 220, 403–413.
Woltering, E.J., deVrije, T., Harren, F., and Hoekstra, F.A. 1997. Pollination
and stigma wounding: Same response, different signal? Journal of
Experimental Botany 48, 1027–1033.
Woltering, E.J., Somhorst, D., and Vanderveer, P. 1995. The role of ethylene
in interorgan signaling during flower senescence. Plant Physiology 109,
1219–1225.
Woodson, W.R., and Brandt, A.S. 1991. Role of the gynoecium in cytokinin-
induced carnation petal senescence. Journal of the American Society for
Horticultural Science 116, 676–679.
Woodson, W.R., Park, K.Y., Drory, A., Larsen, P.B., and Wang, H. 1992.
Expression of ethylene biosynthetic pathway transcripts in senescing
carnation flowers. Plant Physiology 99, 526–532.
Wu, M.J., Zacarias, L., and Reid, M.S. 1991. Variation in the senescence of car-
nation (Dianthus caryophyllus L.) cultivars. II. Comparison of sensitivity
to exogenous ethylene and of ethylene binding. Scientia Horticulturae
48, 109–116.
Wulster, G., Sacalis, J., and Janes, H.W. 1982. Senescence in isolated carnation
petals. Effects of indoleacetic acid and inhibitors of protein ­synthesis.
Plant Physiology 70, 1039–1043.
Xu, X.J., Gookin, T., Jiang, C.Z., and Reid, M. 2007. Genes associated with open-
ing and senescence of Mirabilis jalapa flowers. Journal of Experimental
Botany 58, 2193–2201.
Xu, Y., and Hanson, M.R. 2000. Programmed cell death during pollination-
induced petal senescence in petunia. Plant Physiology 122, 1323–1333.
Xue, J.Q., Li, Y.H., Tan, H., Yang, F., Ma, N., and Gao, J.P. 2008. Expression
of ethylene biosynthetic and receptor genes in rose floral tissues during
ethylene-enhanced flower opening. Journal of Experimental Botany 59,
2161–2169.
Yamada, T., Ichimura, K., Kanekatsu, M., and Van Doorn, W.G. 2007a. Gene
expression in opening and senescing petals of morning glory (Ipomoea
nil) flowers. Plant Cell Reports 26, 823–835.

136
 P H Y SIOLOG Y AN D MOLE C ULAR BIOLOG Y

Yamada, T., Ichimura, K., and Van Doorn, W.G. 2007b. Relationship between
petal abscission and programmed cell death in Prunus yedoensis and
Delphinium belladonna. Planta 226, 1195–1205.
Yamada, T., Ichimura, K., Kanekatsu, M., and Van Doorn, W.G. 2009.
Homologues of genes associated with programmed cell death in a­ nimal
cells are differentially expressed during senescence of Ipomoea nil
­petals. Plant and Cell Physiology 50, 610–625.
Yamada, T., Takatsu, Y., Kasumi, M., Ichimura, K., and Van Doorn, W.G. 2006.
Nuclear fragmentation and DNA degradation during programmed cell
death in petals of morning glory (Ipomoea nil). Planta 224, 1279–1290.
Yamada, T., Takatsu, Y., Kasumi, M., Marubashi, W., and Ichimura, K. 2004.
A homolog of the defender against apoptotic death gene (DAD1) in
senescing gladiolus petals is down-regulated prior to the onset of pro-
grammed cell death. Journal of Plant Physiology 161, 1281–1283.
Yamane, K., and Ogata, R. 1995. Effects of cycloheximide on physiological
parameters of gladiolus florets during growth and senescence. Journal
of the Japanese Society for Horticultural Science 64, 411–416.
Yanagisawa, S., Yoo, S.D., and Sheen, J. 2003. Differential regulation of
EIN3 stability by glucose and ethylene signalling in plants. Nature 425,
521–525.
Yang, S.F., and Hoffman, N.E. 1984. Ethylene biosynthesis and its regulation
in higher plants. Annual Review of Plant Physiology and Plant Molecular
Biology 35, 155–189.
Yangkhamman, P., Fukai, S., and Ichimura, K. 2005. Ethylene production and
vase life of cut carnation flowers under high temperature conditions.
Journal of the Japanese Society for Horticultural Science 74, 337–341.
Yangkhamman, P., Tanase, K., Ichimura, K., and Fukai, S. 2007. Depression
of enzyme activities and gene expression of ACC synthase and ACC oxi-
dase in cut carnation flowers under high-temperature conditions. Plant
Growth Regulation 53, 155–162.

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Chapter 5

Respiratory
Metabolism
Mikal E. Saltveit
University of California, Davis, California

Abstract 140
5.1 Introduction 140
5.2 Why Measure Respiration? 141
5.3 Major Components of Respiration 142
5.3.1 Glycolysis 142
5.3.2 Pentose-Phosphate Shunt 142
5.3.3 Anaerobic Diversion 143
5.3.4 Tricarboxylic Acid Cycle 143
5.3.5 Electron Transport (Chemiosmotic
Phosphorylation) 145
5.4 Measurement of Respiration 146
5.4.1 Loss of Substrate, Heat Production,
and Water 146
5.4.2 Consumption of Oxygen and Production
of Carbon Dioxide 147
5.4.2.1 Static System 147
5.4.2.2 Flow-Through or Dynamic System 149
5.5 Sampling and Analyzing 151
5.6 Instruments and Techniques 152

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5.7 Pre- and Postharvest Factors Affecting Respiration 152

5.7.1 Temperature Effects 152
5.7.2 Respiratory Quotient 154
5.7.3 Physical Stress 154
5.7.4 Internal Factors 155
5.7.4.1 Genotype 155
5.7.4.2 Type of Plant Part 155
5.7.4.3 Respiratory Climacteric 155
5.8 Conclusions and Future Perspectives 156
References 156

Abstract
The three groups of catabolic reactions comprising respiration provide the
energy, the reducing power and carbon fragments used in subsequent ana-
bolic metabolic reactions. Horticultural crops are harvested either at the
optimal stage of maturity and quality (e.g., cucumber, lettuce, zucchini) or
at a stage of maturity that requires subsequent changes to reach maximal
quality (e.g., avocadoes, bananas, tomatoes). In the first group, respiratory
metabolism is suppressed in the postharvest environment to levels only
needed to maintain product quality. In the latter group, the often profound
changes that accompany ripening (e.g., pigment destruction and synthesis,
synthesis of characteristic aroma and flavors, tissue softening, and textural
changes) require significant inputs of energy and substrates derived from
respiration. Measurements of respiration (e.g., carbon dioxide production,
oxygen consumption) give a good indication of the metabolic activity of
the tissue, and thereby the rate of changes in product quality. The rate of
respiration and metabolic activity is tightly coupled to tissue temperature.
Both are reduced by lower product temperatures and increased by elevated
product temperatures. Some commodities (especially those of tropical and
subtropical origin, e.g., avocado, banana, bean, and squash) suffer physi-
ological damage (i.e., chilling injury) if exposed to temperatures below
10°C (50°F) for commodity-specific inductive periods. Respiratory and
metabolic activity can also be reduced by limiting the availability of oxygen
to the tissue. However, too little oxygen induces anaerobic respiration with
production of uncharacteristic compounds that can reduce product quality.

5.1 Introduction
The function of respiratory metabolism is threefold: to extract energy
stored in the chemical bonds of substrate molecules (e.g., glucose, fatty
acids, proteins) and convert it into high-energy phosphate bonds (i.e., ATP),

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to capture reducing power (e.g., in NADH), and to produce small carbon

fragments from which more complex molecules can be synthesized. In
the presence of adequate levels of oxygen, the three sequential respiratory
reactions are glycolysis (i.e., the lysing of glucose to produce pyruvate), the
tricarboxylic acid cycle (i.e., the decarboxylation of pyruvate to CO2 and
production of high-energy compounds, e.g., NADH), and electron trans-
port (i.e., generation of ATP from high-energy compounds by chemiosmotic
phosphorylation). An offshoot of glycolysis, the pentose-phosphate shunt
diverts and reconfigures intermediates from glycolysis into substrates for
subsequent synthetic reactions (e.g., produces the 5-carbon ribose sugars
used in RNA and DNA synthesis). In the absence of oxygen, NAD+ regener-
ated during anaerobic respiration allows glycolysis to continue to extract a
portion of the energy contained in the glucose molecule.
Biological energy is stored and released through redox reactions.
Respiratory metabolism involves the transfer of energy in a substrate mol-
ecule (e.g., glucose) through the extraction and movement of electrons.
Redox (shorthand for reduction–oxidation reactions) involves a coupled
reaction where electrons are gained by a molecule (i.e., it is reduced) while
another molecule is oxidized (i.e., it loses electrons). In aerobic respira-
tion (i.e., in the presence of adequate oxygen), glucose is oxidized (i.e., it
furnishes high-energy electrons) and some of the energy in the electrons
is extracted in the production of ATP as they pass through a number of
enzymatically controlled steps, until they finally reduce oxygen to water
(i.e., transfer the electrons, with the accompanying protons, to oxygen). In
contrast to respiration, photosynthesis reduces carbon dioxide (i.e., adds
electrons to CO2) and oxidizes water (i.e., removes electrons from H 2O) to
produce glucose and oxygen (C6H12O6 and 6O2).
Carbohydrates (i.e., simple sugars like glucose derived from stored
starch) are the main respiratory substrate for most postharvest commodi-
ties. Although glucose contains 686 kcal/mol in chemical bonds, only
673 kcal/mol is available to do “work” because 13 kcal/mol is lost due to the
increase in disorder of the products (i.e., an increase in entropy). However,
only 281 kcal/mol of the 673 kcal/mol (42%) is captured in the 38 ATP
(7.4 kcal/mol) formed because of the inefficiencies of phosphorylation.

5.2  Why Measure Respiration?

Because respiration is tightly controlled so that no more ATP is produced
than is needed for metabolism, and because ATP production is tightly cou-
pled to respiration, the rate of respiration is normally a very accurate indica-
tor of the metabolic activity of the tissue. We do not measure respiration to
find only the rate of CO2 production or O2 consumption, but also the under-
lying rate of metabolism in the tissue. The rate of metabolism is usually

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inversely related to the shelf life of the commodity. The higher the rate of
respiration, the more quickly there will be a decrease in the quality, nutri-
tion, or taste of the commodity. Since we cannot easily measure changes in
these parameters of shelf life, we use measurements of the rate of respira-
tion as their surrogate.

5.3  Major Components of Respiration

5.3.1 Glycolysis
As its name implies, the 10 enzymatic reactions in glycolysis break (or lyse)
the 6-carbon glucose molecule into two 3-carbon pyruvate molecules. The
reactions of glycolysis take place in the cytoplasm. Glucose is first phosphory-
lated in a series of reactions by two ATPs to produce fructose-1,6-diphosphate.
Living cells are homeostatic over short periods of time and tend to maintain
the status quo by carefully regulating the concentration of ATP. A key enzyme
in glycolysis is phosphofructokinase (PFK), a multicomponent enzyme that
converts fructose-6-phosphate into fructose-1,6-diphosphate, thereby pre-
paring the hexose for cleavage into two dissimilar phosphate-containing
3-carbon molecules (i.e., dihydroxyacetone phosphate and glyceraldehyde-
3-phosphate [G3P]), which are interconvertible. The rate of glycolysis is reg-
ulated primarily through controlling the activity of PFK, an enzyme that is
inhibited by ATP, one product of respiration. However, almost every enzyme
in the respiratory pathway is under some sort of metabolic control, indicating
the importance of careful regulation of respiration and metabolism.
Each molecule of G3P undergoes a series of reactions during which
one molecule of NADH and two molecules of ATP are produced. The result-
ing two molecules of the 3-carbon pyruvate contain all six carbons that were
in the original molecule of glucose. Two ATPs were needed to initiate the
reaction, but four ATPs and two NADHs were produced, for a net gain of
two ATPs and two NADHs. So while there are the same numbers of carbon
atoms in the two pyruvate molecules as there were in the original glucose
molecule, the pyruvate molecules have lost about 20% of the extractable
energy originally present in the glucose molecule. The remaining 80% is
extracted by the two subsequent reactions of respiration. Notice that no oxy-
gen was consumed, and no carbon dioxide was produced during glycolysis.

5.3.2  Pentose-Phosphate Shunt

Glucose-6-phosphate can be diverted from glycolysis to another oxidative
pathway that generates NADPH and five carbon substrates for further bio-
synthesis (e.g., ribose-5-phosphate for RNA synthesis).

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 Respir ato ry Me ta bo lism

There are two similar forms of nicotinamide adenine dinucleotide:

one with a phosphate group (NADPH) and one without a phosphate group
(NADH). In general, NADH participates in catabolic reactions, that is, reac-
tions of cellular respiration that break down molecules to release energy,
while NADPH participates in anabolic reactions, such as carbon fixation
during photosynthesis, that is, reactions that consume energy in order to
build up or synthesize larger molecules.

5.3.3  Anaerobic Diversion

NAD+ is needed for glycolysis to function. In the absence of oxygen (i.e.,
anaerobic), NAD+ accumulates in its reduced state (NADH) and all energy
production ceases with the subsequent death of the cell unless the oxidized
form (NAD+) can be regenerated. Anaerobic respiration starts with the
enzyme pyruvate decarboxylase removing a CO2 from pyruvate, forming
the 2-carbon acetaldehyde. However, acetaldehyde is a very reactive mol-
ecule and is quickly converted to ethanol by the enzyme alcohol dehydroge-
nase with the concomitant regeneration of NAD+. In some cells, especially
those that are less acidic, the enzyme lactate dehydrogenase regenerates
NAD+ during the production of lactic acid. However, the accumulation of
lactic acid acidifies the cell and stimulates the activity of pyruvate decar-
boxylase, which thereby shifts carbon flow from the production of lactate
to the production of ethanol. Differences in these two pathways are illus-
trated by the production of yogurt and beer. Lactic acid production during
fermentation to produce yogurt occurs without the release of CO2, while
production of ethanol during beer production is accompanied by the release
of CO2, which generates the effervescent beverage.

5.3.4  Tricarboxylic Acid Cycle

Pyruvate produced by glycolysis can enter a cyclic series of eight reactions
called the tricarboxylic acid cycle (TCA) (also called the citric acid cycle or
Krebs cycle) (Figure 5.1). Hans A. Krebs was the scientist who first eluci-
dated the cycle, and the first molecule in the cycle is citric acid, a tricarbox-
ylic acid. These reactions take place in the liquid portion (i.e., the matrix)
of the mitochondria, a semiautonomous organelle within eukaryotic cells.
Pyruvate is decarboxylated, yielding a molecule of CO2 and a 2-­carbon
acetate residue that is attached to a co-enzyme forming acetyl-CoA. The
2-carbon acetate group is then transferred to the 4-carbon oxaloacetate
molecule, forming the 6-carbon citric acid. As the cycle progresses, an
additional two molecules of CO2 are released, and the resulting 4-carbon
molecule is rearranged into the starting product of oxaloacetate, thereby

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Polysaccharides
Proteins Fats
Simple sugar
(e.g., glucose)
Amino acids Fatty acids
glycerol

Glycolysis
-oxidation

CO2
Pyruvate

Acetyl CoA

Citric
acid CO2
cycle
CO2

Reducing Power
3 O2 (NADH, FADH2)

Electron transport

ATP ADP + Pi
3 H2O Mitochondria Cytoplasm

Figure 5.1  Overview of important features of cellular respiration.

completing the cycle. During the cycle, three NADHs, one FADH2, and one
ATP are produced from each pyruvate molecule (Figure 5.2).
High-energy molecules are not the only product of the TCA cycle. The
intermediates within the cycle can be substrates for subsequent synthetic
reactions. For example, pyruvate is used in the synthesis of acetaldehyde,
ethanol, lactic acid, and alanine; acetyl CoA is used in the synthesis of fatty
acids, cuticular compounds, isoprenoids, carotenoids, sterols, terpenes,
and aromatic amino acids; α-ketoglutarate is used in the synthesis of glu-
tamic acid, other amino acids, chlorophyll, cytochromes, and phytochrome;

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 Respir ato ry Me ta bo lism

Pyruvic acid
COOH Pyruvic
C=O decarboxylase CO2
NADH
CH3
Lactate Acetaldehyde
dehydrogenase Pyruvic H-C=O
2H+ oxidase CH3 NADH
+
NAD
Alcohol
CO2 dehydrogenase
COOH S-CoA 2H+
COOH C=O +
NAD
H-C-OH CH3 H
CH3 H-C-OH
Acetyl-CoA CH3
Lactic acid

Ethanol
To TCA cycle

Figure 5.2  The fate of pyruvic acid in respiratory metabolism.

and oxaloacetic acid is used in the synthesis of aspartic acid, alkaloids, and
nucleic acids.
Notice that in the TCA cycle, all the carbon from the glucose molecule
was released as CO2, but no oxygen has yet been consumed.

5.3.5  Electron Transport (Chemiosmotic Phosphorylation)

The preceding respiratory reactions have converted all the carbon in the
original glucose molecule into CO2, but in none of the previous reactions has
oxygen been consumed. The final reaction of respiratory metabolism regen-
erates NAD+ and converts the energy contained in NADH into ATP. During
this process, the electrons contained in NADH and FADH2 are transferred
to oxygen, the terminal electron acceptor, thereby producing water.
It was previously thought that the series of reactions generating ATP
from NADH (and FADH2) in the mitochondria were a series of coupled enzy-
matic reactions, with the product of one reaction furnishing the substrate
for the next reaction. It is now known that the production of ATP involves
using the high-energy electrons from NADH to pump protons across the
inner mitochondrial membrane, thereby establishing a proton gradient
between the intermembrane space and the matrix. The protons reenter the
matrix by passing through an H+ -ATPase complex embedded in the inner

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membrane. This ATPase uses the energy in the returning protons to pro-
duce ATP from ADP and Pi (inorganic phosphate), a process called chemi-
osmotic phosphorylation.

5.4  Measurement of Respiration

Since the rate of respiration is so tightly coupled to the rate of cell metabo-
lism, measurements of respiration can afford an easy, nondestructive means
of monitoring the metabolic and physiological state of the tissues. Nowhere
in plant science is this truer than in the area of postharvest physiology,
where the events of senescence and ripening are often signaled by abrupt
changes in respiratory behavior. For this reason, postharvest physiologists
have spent considerable time devising means to measure respiration.
The overall reactions of respiratory metabolism consume substrate
molecules (i.e., carbohydrates, fatty acids, proteins, and oxygen) and pro-
duce high-energy compounds, CO2, and water. The rate of respiration could
be measured by the rate at which the substrates disappear or the products
appear. Apart from the water produced by respiration, which is relatively
trivial compared to the very high water content of plant (and particularly
fruit) tissues, all the substrates and products of respiration have been used
in determining respiration rate. They are: loss of substrate, loss of oxygen,
increase in carbon dioxide, and production of heat.
C6H12O6 + 6O2 → 6CO2 + 6H 2O + 38 ATP + 392 kcal (heat)

5.4.1  Loss of Substrate, Heat Production, and Water

It is theoretically possible to monitor the respiration of stored commodities
by measuring the loss of dry matter. During extended storage periods at
warm temperatures, the loss of dry matter of a respiring commodity can
be substantial. When glucose (a hexose sugar) is the substrate, 180 g of
glucose is lost for each 264 g of CO2 produced by the commodity. The rate
of dry weight loss (i.e., loss of glucose) can be estimated by multiplying
the weight of CO2 produced by 0.68: the ratio of the molecular weight of
glucose (180) divided by the molecular weight of the six CO2 produced
(264 = 6 × 44).
In addition to actual dry weight loss (which could be an important
consideration, for example, in onions being stored prior to dehydration), the
oxidation of substrates can be an important postharvest loss because of the
reduction in food value of the commodity to the consumer, reduced taste
quality (sweetness), and potential for acceleration of senescence in tissues
whose storage reserves have been depleted.

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 Respir ato ry Me ta bo lism

The rate of respiration has occasionally been measured by determin-

ing the rate at which heat is produced by the commodity. All of the heat
produced during respiration is lost from the tissue either by conduction to
the cold storage room atmosphere or by vaporization of water in the tissue.
If all the energy not captured in ATP (673 − 281 = 392 kcal) was lost from
the tissue by vaporizing water, 676 g of H 2O would be lost for each 180 g of
glucose oxidized (392 kcal/580 cal/g H 2O). Since 108 g of H 2O is produced
during respiration, there would be a net loss of 568 g of H 2O. This loss will
occur even in a water-saturated atmosphere. Apples at 0°C have a respira-
tion rate of about 4 mg CO2/kg-h, so 10 kg of apples would respire 1.2 g of
CO2, or about 0.82 g of glucose would be lost in a month (30 days). During
this time, 1.78 kcal of heat would have been produced [(1.2/264) × 392] and
could have led to the loss of 3.1 g of water (1.78 kcal/580 kcal/g H 2O).
Measurement of heat production is also important when engineer-
ing refrigeration systems used during postharvest handling and storage.
Calculation of heat production from the respiration equation shows that pro-
duction of 1 mg of CO2 yields 2.55 cal. In the language of the refrigeration
­engineer, a respiration rate of 1 mg CO2/kg-h indicates a heat production of
61.2 kcal/metric ton/day (220 BTU/tonne of produce/day). The British ther-
mal unit (BTU) is defined as the heat required to raise 1 lb of water by 1°F.

5.4.2  Consumption of Oxygen and

Production of Carbon Dioxide
Almost all practical methods for measuring the respiration of plant tis-
sues involve the estimation of uptake of O2 or release of CO2 by the tissue.
These methods are generally simple and have the great advantage of being
nondestructive. Indeed, they have often been used on commodities still
attached to the plant. Two general systems have been used for measuring
gas exchange: the static and the flow-through system.
In either system, respiration rates are usually expressed as weight
or volume of gas produced or consumed per kilogram of fresh weight of
product per hour, for example, 125 mg CO2/kg fresh wt/h or 25 ml O2/kg
fresh weight/h.

5.4.2.1  Static System

In the static system, tissue is placed in a sealed container and the accumu-
lation of CO2 or the depletion of O2 in the atmosphere is periodically mea-
sured (Figure 5.3). This method can be employed over brief periods of time,
especially with even very small portions of tissue (e.g., meristems, florets,
buds), but it has the disadvantage of being a nonequilibrium system. The
depletion of O2 and accumulation of CO2 or other gases (particularly C2H4)

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0.4

O2 CO2

Carbon dioxide %
0.3

0.2

0.1

0
0 1 2 3 4 5
Time
(a) (b)

Figure 5.3  Representation of the setup for measuring O2 consumption or

CO2 production in a static or closed system. (a) Depiction of a commodity in
a closed container. (b) Graph showing the increase in CO2 that would occur
as the commodity respired in a closed container. Note the derivation from
linearity as the CO2 concentration exceeds around 0.2%.

may strongly affect the tissue and its respiration rate. As can be seen in the
graph, the accumulation of CO2 becomes nonlinear at around 0.2%.
Problems with the accumulation of physiologically active concen-
trations of CO2 and C 2H4 can be overcome by including KOH to absorb
CO2 and KMnO 4 to oxidize C 2H4 . Indeed, some respirometers ( jargon for
a machine that measures respiration) measure the pressure (or volume)
change resulting from absorption of respired CO2 . The most notable of
these, the Warburg respirometer, was instrumental in determining the
pathway of glycolysis and has driven many postharvest physiologists to
strong drink because of the technical difficulties often encountered in
its use.
To calculate the rate of respiration in a static system, you need to know
the following: volume of container, weight of tissue, initial carbon dioxide
concentration, length of time, and final carbon dioxide concentration.
For example, assume that a 280 g apple is enclosed in a 1560 ml
container for 15 min. During that time, the CO2 concentration in the jar
increased from 0.03% to 0.23%, or the CO2 concentration increased 0.2%.
Converting the weight and time units to kilograms and hours gives a
weight of 0.28 kg and a sampling period of 15/60 = 0.25 h. The simplest
way to calculate the amount of CO2 produced during the sampling period
would be to multiply the difference in CO2 concentrations between the
beginning and end of the sampling period (i.e., 0.2%) by the total volume of
the container (0.2% × 1560 ml = 3.12 ml). Dividing that volume of CO2 by

148
 Respir ato ry Me ta bo lism

the weight (in kg) and the sampling period (in hours) would give the rate
of CO2 production (i.e., 3.12 ml/(0.28 kg × 0.25 h) = 44.6 ml CO2/kg-h).
However, this calculation could be criticized because many assump-
tions were made that can significantly alter the answer. For example, it did
not take into account the fact that the volume occupied by the apple reduced
the total gas volume in the jar; i.e., the density of the tissue could deviate
from 1 and affect its volume, and the solubility of the gas being measured
in the cellular solution. The density of an apple is around 0.8 g/ml, so the
volume occupied by the apple would be 280/0.8 = 350 ml. The liquid in the
apple fruit will absorb some of the CO2 produced. The solubility of 100% CO2
in water at 20°C is 0.878 ml CO2/ml water (and increases with decreasing
temperatures). Taking all these caveats into consideration, we can conclude
that under normal physiological conditions, the rate of CO2 production can
be calculated to within about 5% of its true value by ignoring the volume
changes introduced by the tissue. The volume of CO2 produced can be cal-
culated by multiplying the change in concentration by the entire void vol-
ume of the container.
The significantly lower solubility of oxygen and ethylene in the
aqueous solution of most commodities requires that the volume of the
tissue be subtracted from the container volume when calculating their
production rate.

5.4.2.2  Flow-Through or Dynamic System

In the flow-through system, tissue is placed in a sealed container through
which a stream of gas flows (Figure 5.4). This flow must be accurately main-
tained during the experiment. After a period of time that is usually equal to
the time required for the flow to replace the container volume three times,
the difference in gas concentrations between the inlet and outlet flows
should be nearly equal to the gases being consumed or produced by the tis-
sue. The system is then in equilibrium, and the differences in concentration
between the inlet and outlet gases will be equal to their production (e.g.,
CO2, C2H4) or consumption (e.g., O2) by the tissue. Gas samples can then
be taken for analysis.
The flow-through system is well suited for long-term experiments and
controlled atmosphere experiments when atmospheres of various composi-
tions are used to ventilate the container. This system has the added advan-
tages that leaks are not critical as long as the gases in the container are
completely mixed, and the physiological active concentrations of evolved
gases can be kept below their critical levels.
To calculate the rate of respiration in a dynamic system, you need to
know the following: weight of tissue, inlet carbon dioxide concentration,
flow of gas, and outlet carbon dioxide concentration.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Gas in

0.15

O2 CO2

Carbon dioxide %
0.10

0.05

0.00
0 2 4 6 8 10
(a) (b) Time
Gas out

Figure 5.4  Representation of the setup for measuring O2 consumption or

CO2 production in a dynamic or flow-through system. (a) Depiction of a com-
modity in a flow-through system. (b) Graph showing the increase in CO2 that
would occur as the commodity respired in flow of gas. Note how the CO2
concentration increases to a steady-state level as the loss of CO2 from the
container approaches the production of CO2 by the commodity. Judicious
selection of commodity weight and flow should keep the equilibrium CO2
concentration below around 0.2%.

The production of carbon dioxide or ethylene or the consumption of

oxygen is calculated by multiplying the increase (or decrease) in concen-
tration between the inlet and outlet tubes by the flow rate. Dividing by the
fresh weight (in kg) of the tissue gives the rate of carbon dioxide production
in ml/kg-h.
For example, assume that a 280 gram apple is enclosed in an 843 ml
jar. The carbon dioxide concentration increases from 0.03% to 0.23% as air
flows through the jar at 100 ml/min. It usually takes three volume changes
for a flow-through system to come into equilibrium, so it will take 25.3 min-
utes [(3 × 843 ml)/100 ml/min] for this system to come into equilibrium.
The concentration of carbon dioxide increased 0.20%, so the amount of car-
bon dioxide produced per hour is 12.0 ml (i.e., 0.2% × 100 ml/min × 60
min/h). Dividing by the 0.28 kg fresh weight gives a rate of production of
42.8 ml CO2/kg-h [i.e., (12.0 ml CO2/h)/0.28 kg].
The accuracy of the dynamic method is dependent on knowing the
exact flow rate. The container volume and flow rate are both important in
determining the time it will take for the system to come to equilibrium, but

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 Respir ato ry Me ta bo lism

as long as the system is at equilibrium, the volume of the container is not

relevant to the calculations. Small leaks are also not important as long as
the gases are thoroughly mixed within the container.

5.5  Sampling and Analyzing

Gas samples can be taken with gastight plastic syringes by inserting the
syringe needle through a septum in the container walls or into flexible tub-
ing protruding from the container. The material used to make most of the
glass and plastic sampling syringes is very impermeable to carbon dioxide
and oxygen, so samples can be held in these containers for days without
significant contamination by gases in the surrounding atmosphere. The
samples are injected in an analyzer and the output compared to standards
to arrive at the correct gas concentration in the sample.
Qualitative and quantitative measures of the component gases in the
sample can be done with a variety of methods and instruments. Early meth-
ods of gas analysis relied on changes in the volume of the gas sample when
specific gases were absorbed. These methods are only of historical interest
today. However, some of the early colorimetric methods are still used. The
reaction of a specific gas with a reagent immobilized on a solid support
causes a color change that identifies the gas and can be used in certain
configurations to estimate the concentration of the gas.
Modern methods of gas analyses rely on electronic instruments.
Some instruments are designed to measure a specific gas. For example, the
infrared analyzer is specific for carbon dioxide, and the paramagnetic or
electrochemical analyzer is specific for oxygen. These instruments do not
require the separation of a gas sample into its individual gas components.
The gas chromatograph separates a gas sample into its individual com-
ponents by passing the sample down a long tube filled with a coated solid
support. The sample is injected into a stream of inert gas (usually nitrogen)
that carries the sample down the tube. The coating and support are selected
to achieve the desired separation of the gases. Upon emerging from the tube,
the separated gases pass through a detector that measures some property
of the gas. Thermal conductivity detectors measure the capacity of the ­sample
to remove energy from a heated filament. Ionization detectors measure the
electrical conductivity of the gas after it has been ionized by ­combustion
(flame ionization detector) or irradiation (photoionization detector).
Samples need not be drawn if the respired gases can be collected. In
a dynamic—or flow-through—system, the commodity is placed in a sealed
container through which a stream of gas is passed. The exit stream is passed
through a column containing caustic solutions or granules that absorb the
respired carbon dioxide. Respiration is determined by subsequent titrimetric

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or gravimetric analysis of the material in the column. This system is r­ elatively

easy to set up and is independent of airflow rates, but any leaks in the tubes
leading from the respiration container (which is usually impossible to seal
completely) cause erroneous results. This method is rarely used because
large amounts of plant tissue and long sampling times are required, and the
method is not as accurate as the instrumental methods described previously.

5.6  Instruments and Techniques

Gas chromatograph: All respiratory gases and ethylene can be
assayed with a gas chromatograph by selecting the proper column
and detector. The instrument can be set up to use small (< 1 ml)
samples and rapidly (< 1 min) assay them.
Infrared CO2 analyzer: An instrument that specifically mea-
sures CO2. It can be modified to analyze small (< 1 ml) samples
within 10 s.
Paramagnetic O2 analyzer: An instrument that specifically mea-
sures oxygen. It requires a large sample of gas to analyze and has a
slower response than the other two instruments. Oxygen can also
be measured by polarography or electrochemical means.
Kitagawa tubes: These are glass tubes filled with a specific for-
mulation of reagent to detect a specific gas. A known volume of gas
(e.g., 100 ml) is drawn through the tube, and the reaction between
the reagents and the specific gas component produces a colored
reaction. The length of the colored band is proportional to the con-
centration of the gas in the sample.
Pressure or volume changes: If CO2 produced by respiration
is absorbed by a chemical (e.g., KOH) in a closed container, the
reduction in the volume of O2 will be measurable as a decrease in
pressure. A monometer can indicate the change in pressure by a
small change in volume of a liquid in a U-tube. A large gas sample
(e.g., 100 ml) is usually required, and minor temperature changes
can significantly change the pressure in a sealed container.

5.7  Pre- and Postharvest Factors

Affecting Respiration

5.7.1  Temperature Effects

Without a doubt, the most important factor affecting the rate of respiration
is temperature. Over the physiological range (i.e., 0°C−30°C), increased
temperatures cause an exponential rise in respiration; in general, the

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velocity of a biological reaction increases two- to three-fold for every 10°C

rise in temperature.
The temperature coefficient for a 10°C interval is called the Q10,
which can be calculated by measuring the rate of the respiration (e.g., CO2
production) at two temperatures. Obviously, if the temperature difference
is 10°C, then the Q10 will simply be the quotient of the two rates, that is,
Q10 = R2/R1.
The temperature coefficient is a useful concept, because it allows
calculation of expected respiration rates at one temperature from a known
rate at another temperature using the equation given above. However, the
respiration rate in perishable commodities does not follow ideal behavior,
and the Q10 can vary considerably. At higher temperatures, the Q10 is usu-
ally smaller than at lower temperatures. Typical figures for Q10 are given in
Table 5.1.
These typical Q10 values allow us to construct a table showing the
effect of different temperatures on the rates of respiration or deterioration
and relative shelf life of a typical perishable commodity (Table 5.2).
Table 5.2 shows that if a commodity has a mean shelf life of 13 days at
20°C, it may be stored for as long as 100 days at 0°C and will last no more
than 4 days at 40°C.
Although respiration is normally reduced at low, but nonfreezing tem-
peratures, certain commodities, chiefly those originating in the tropics and
subtropics, show abnormal respiration when the temperature falls below
10°C−12°C. Typically, the Q10 is much higher at these low temperatures

Table 5.1  Typical Q10 Values

Temperature Range (°C) Q10
  0 – 10 2.5 – 4.0
10 – 20 2.0 – 2.5
20 – 30 1.5 – 2.0
30 – 40 1.0 – 1.5

Table 5.2  Effect of Temperature on Rate of Deterioration

Assumed Velocity of Relative Shelf
Temperature (°C) Q10 Deterioration Life
 0  1.0 100
3.0
10  3.0 33
2.5
20  7.5 13
2.0
30 15.0  7
1.5
40 22.5  4

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than predicted; that is, the rate of respiration falls abnormally rapidly. In
some cases, the respiration rate may even increase dramatically at lower
temperatures, possibly associated with increased glycolysis to compensate
for loss of mitochondrial oxidation. These events are symptoms of the onset
of chilling injury, an economically important low-temperature phenomenon.
Another important respiratory effect of exposure of sensitive com-
modities to chilling temperatures is abnormally high respiration rate (sus-
tained if the tissue has been irreversibly damaged) when the commodity is
returned to normal temperatures. This enhanced respiration presumably
reflects the cells’ efforts to detoxify metabolic intermediates that may have
accumulated during the chilling exposure, as well as to repair damage to
membranes and other subcellular structures.
As the temperature rises beyond the physiological range, the rate
of increase in respiration falls. It becomes negative as the tissue nears its
thermal death point, when metabolism is disorderly and enzyme proteins
are denatured. Many tissues can tolerate high temperatures for short times
(e.g., minutes), and this property is used to advantage in killing some surface
fungi in stone fruits and papaya. Continued exposure to high temperatures
causes phytotoxic symptoms and then tissue collapse. However, condition-
ing and heat shocks (i.e., short exposures to potentially injurious tempera-
tures) can modify the tissue’s responses to subsequent harmful stresses.

5.7.2  Respiratory Quotient

When carbohydrates are the primary substrate respired to produce ATP,
there is an equimolar production of CO2 to O2 consumed. The proportion of
CO2 produced to O2 consumed will vary when other substrates are respired.
Because organic acids (e.g., citric acid) are the product of partially oxidized
glucose, tissue using organic acids in respiration will produce more CO2
per O2 consumed. The ratio of CO2 produced to O2 consumed is called the
­respiratory quotient (RQ) and is defined as the ratio of CO2 evolved to O2 con-
sumed (measured in moles or volumes). RQ values range from 0.7 to 1.3 for
aerobic respiration: carbohydrates have an RQ of around 1.0, while lipids have
an RQ of less than 1.0, and organic acids have an RQ of greater than 1.0. In
the aerobic range, calculation of the RQ can indicate which substrate is being
oxidized during respiration. Very high RQ values can indicate that anaerobic
respiration may be occurring in some parts of the tissues under examination.

5.7.3  Physical Stress

Physical stress has a pronounced effect on the respiration of perishable
products. Both biotic and abiotic stresses cause the tissue to produce a

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signal that migrates from the site of injury and induces a wide range of
physiological changes in adjacent, nonwounded tissue. Some of the more
important changes include enhanced respiration, ethylene production,
phenolic metabolism, and wound healing. Most wound responses are det-
rimental to quality (e.g., rapid softening, browning, toughening), but some
(e.g., wound healing in potatoes) are necessary for their long-term storage.

5.7.4  Internal Factors

A number of commodity parameters are directly related to the respiration
rate and respiratory pattern.

5.7.4.1 Genotype
Some fruits are more perishable than others. You would not, for example,
expect a strawberry to last as long as apples in your fruit bowl. This relative
perishability is usually reflected in the respiration of the different commodi-
ties. The respiration of strawberries is much higher than that of apples.
Some short-lived apple and melon fruit are from cultivars with higher rates
of respiration than long-lived cultivars.

5.7.4.2  Type of Plant Part

There is a very large variation in the respiration rate among different plant
organs. Storage tissues such as nuts and tubers have low respiration rates.
Tissues with vegetative or floral meristems, such as asparagus and broc-
coli, have very high respiration rates.
As plant organs mature, their rate of respiration typically declines.
This means that commodities harvested during active growth, such as
many vegetables and immature fruits, have high respiration rates. Mature
fruits, dormant buds, and storage organs have relatively low respiration
rates. After harvest, the respiration rate typically declines, slowly in non-
climacteric fruits and storage organs, and rapidly in vegetative tissues and
immature fruits. The rapid decline presumably reflects depletion of respi-
rable substrates that are typically low in such tissues.

5.7.4.3  Respiratory Climacteric

An important exception to this general decline in postharvest respiration
is the rapid and sometimes dramatic rise in respiration during the ripen-
ing of climacteric fruit. This rise, which has been the subject of intense
study for many years, is normally considered as having four distinct phases:
(1)  preclimacteric minimum, (2) climacteric rise, (3) climacteric peak,
and (4) postclimacteric decline. The division of fruits into climacteric and

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nonclimacteric types has been very useful for postharvest physiologists.

However, some fruits, for example, kiwifruit and cucumber, appear to blur
the distinction between the groups. Respiratory rises also occur during
stress and other developmental stages, but a true climacteric only occurs
coincident with ripening.

5.8  Conclusions and Future Perspectives

Respiratory metabolism during postharvest handling and storage should
be carefully regulated to proceed at the minimal rate that maintains prod-
uct quality. While elevated metabolic rates are sometimes needed for short
periods of time to increase quality (e.g., ripening of climacteric fruit, ­curing
of root vegetables), the subsequent maintenance of product quality is opti-
mized at far lower metabolic rates. Storing, transporting, and marketing
harvested horticultural commodities at their lowest tolerated temperature
is the primary method used to suppress metabolism and maximize quality
retention. Controlled and modified atmospheres are used to extend stor-
age life by reducing the oxygen concentration and increasing the carbon
dioxide concentration within the commodities. Postharvest practices could
be formulated that increase the  tissue’s tolerance to lower oxygen and
higher carbon dioxide levels. The significant rise in respiratory metabolism
that accompanies the ripening of climacteric fruit may be physiologically
­unnecessary. Elimination of the respiratory climacteric by either genetic
manipulation or postharvest treatments would significantly increase the
retention of ­storage reserves that could possibly contribute to quality
preservation.

References
Kader, A.A. 1992. Postharvest Technology of Horticultural Crops. 2nd ed.
Division of Agriculture and Natural Resources, University of California,
Oakland.
Kader, A.A., and Saltveit, M.E. 2003. Respiration and gas exchange. In
Postharvest Physiology and Pathology of Vegetables, ed. J.A. Bartz and J.K.
Brecht. Marcel Dekker, New York, pp. 7–30.
Kays, S.J., and Paull, R.E. 2004. Postharvest Biology. Exon Press, Athens, GA.
Wills, R.B.H., McGlasson, B., Graham, D., and Joyce, D. 1998. Postharvest:
An Introduction to the Physiology and Handling of Fruit, Vegetables and
Ornamentals. Wallingford, Oxon, UK: CAB International.

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Chapter 6

Stomata and
Postharvest Physiology
Uulke van Meeteren1 and Sasan Aliniaeifard 2
1Wageningen University, Wageningen, the Netherlands
2University of Tehran, Tehran, Iran

Abstract 158
6.1 Introduction 158
6.2 Stomata 159
6.2.1 Role of Stomata in Plants 159
6.2.2 Mechanism of Stomatal Closure and Opening 161
6.2.3 Signal Transduction Pathways in Guard Cells
for Stomatal Closure 164
6.3 Role of Stomata in the Postharvest Phase 166
6.3.1 Stomata in Relation to Vase Life of Cut Flowers 166
6.3.2 Stomata in Relation to Quality of Vegetables 169
6.3.3 Stomata in Relation to Quality of Fruits 172
6.4 Preharvest Conditions Leading to Postharvest
Problems via Stomata 174
6.4.1 Relative Humidity and Stomata Control 175
6.4.1.1 How Does Preharvest Low VPD Affect
Postharvest Stomata Control? 176
6.4.1.2 Induction of Stomata Morphological
Changes by Low VPD 179

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6.4.2 Temperature 181

6.4.3 Light 182
6.5 Stomata and Tolerance to Postharvest Diseases
and Physiological Disorders 184
6.6 Postharvest Treatments and Stomata 186
6.7 Ethylene, Stomata, and Senescence 187
6.8 Conclusions and Future Perspectives 190
References 191

Abstract
Stomata are pores in the gastight waxy cuticula that covers the outer surface
of aerial parts of plants. They make uptake of CO2 possible, which is needed for
photosynthesis. At the same time, water vapor will leave the plant via the sto-
mata. To optimize photosynthesis, while at the same time preventing excess
water loss, stomata control their opening by a signaling network of pathways
that respond to environmental conditions such as light and darkness, water
vapor pressure deficit (VPD), temperature, CO2, and ethylene. Water loss is
one of the most obvious changes in harvested vegetables, often limiting mar-
keting life, and a negative water balance (uptake of water is insufficient to
compensate transpiration) is one of the most important reasons for the end of
the vase life of cut flowers. Also, the postharvest quality of some fruits is nega-
tively affected by water loss. In leafy vegetables, stomata are portals that make
invasion of bacteria into the inner tissue possible and protect in that way bac-
teria for sanitizers in washing solutions with the risk of food-borne bacterial
diseases. This chapter discusses how several environmental factors, during
preharvest cultivation as well as postharvest storage, influence stomata clos-
ing control in harvested cut flowers, vegetables, and fruits. Also, the role of the
number of stomata and their variability between genotypes and due to cultiva-
tion conditions are discussed in relation to postharvest life. One of the most
striking factors is low VPD (high humidity) during the growth of plants: after
exposure of several days to low VPD, the control of stomata closure is largely
disturbed; stomata do not respond anymore to stimuli that normally induce
closure. This malfunctioning is very persistent and results in high water loss
afterward in the harvested products. Fast cooling of produce can close the
stomata of some crops, while in others, the stomata stay open until wilting.

6.1 Introduction
A waxy cuticle covers the outer surface of the epidermal cells of the a­ erial
parts of plants. This cuticular layer protects plants from drying via its
airtight properties. However, a plant needs gas exchange (intake of CO2

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and O2) for photosynthesis and respiration. Therefore, there are pores in the
cuticle, called stomata, of which the opening can be controlled by surround-
ing guard cells. Stomata are the main openings connecting the internal
leaf space to the outside environment. The intercellular air spaces in plant
tissues are saturated with water vapor, which will exit the tissue through
these stomata. Water loss and respiration are two of the most important
processes involved in postharvest physiology of freshly harvested products;
therefore, understanding the control of the opening of the stomata in the
postharvest phase is of importance to control the postharvest behavior of
plant produce.
Besides gas exchange, stomata also offer opportunities for fungi
and bacteria to enter plant tissues. The infection can take place during the
preharvest growth period as a latent infection that results in postharvest
losses, but contamination with bacteria can also take place during handling
after harvest. Especially when human pathogenic bacteria enter plant tis-
sue via the stomata, they are a serious threat to human health.
The closing and opening behavior of stomata is instantly affected by
many environmental cues. Above this, exposure for some time to specific
environmental conditions can influence the stomatal closing control after-
wards. As a consequence, climatic conditions during cultivation can affect
the stomatal closing control during the postharvest phase. In this chapter,
we describe the control mechanism of stomata control, how this control can
be affected by previous environmental conditions, and the specific possible
consequences for quality changes during the postharvest phase of orna-
mentals, vegetables, and fruits.

6.2 Stomata

6.2.1  Role of Stomata in Plants

The presence of a waxy layer (cuticle) that covers the outer surface of the
epidermal cells greatly reduces water loss from the epidermal cell walls
to the surrounding atmosphere. It also blocks exchange of other gases
between the plant tissue and the air around the plant. Stomata are pores in
the plant epidermis surrounded by a pair of kidney-shaped cells in dicots
(and some monocots) or dumbbell-shaped cells in monocots, the guard
cells (Sack, 1987). These stomata provide an entry channel for CO2 and
an exit for water vapor; this last process is called transpiration. In dicots,
the pattern of stomatal arrangement over the leaf surface is irregular,
while their arrangements are in parallel rows in monocots (Figure 6.1).
The guard cells regulate the opening and closing of stomata to control gas
exchange between a plant and the surrounding environment. The main role

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(a)

(b)

Figure 6.1  Distribution of stomata over the leaf of Tradescantia virginiana

as an example of monocots (a) and Vicia faba as an example of dicots (b).
(Pictures are from author’s archive.)

of stomata is providing enough CO2 for photosynthesis, while at the same

time protecting the water status of the plant by preventing excess water loss
via its opening. They have the capacity of adaptation to the environmental
conditions to minimize water loss while promoting the acquisition of CO2.
Therefore, stomata and their opening and closing control are of vital impor-
tance for the survival of plants. Because of this vital importance, evolution
has resulted in a complex signaling network of pathways that will trigger
opening or closing under certain environmental conditions. This signaling
system comprises positively as well as negatively (inhibiting) interacting
modules (Sirichandra et al., 2009).
Besides photosynthesis and water loss, the temperature of a plant
plays a vital role in plant functioning and survival. High leaf temperatures
can decrease photosynthesis, cause damage to the components of the pho-
tosynthetic apparatus (Schreiber and Berry, 1977; Wise et al., 2004; Camejo
et al., 2005), or ultimately result in the death of tissue. Leaf temperature
depends mainly on the energy going into the leaf (absorbed solar irradia-
tion, absorbed infrared irradiation from surroundings) and energy going
out of the leaf (emitted infrared radiation, heat convection, heat conduc-
tion, heat loss accompanying water evaporation). Because of the role of this
last process, leaf temperature is affected by relative air humidity (RH), leaf

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boundary layer conductance, stomatal conductance, and leaf morphologi-

cal traits, such as the presence of trichomes on the leaf (Jones, 1999; Nobel,
1999). Energy is released when water evaporates at the leaf cell walls to
the intercellular spaces. Energy is required to break down the hydrogen
bonds in the liquid phase of water molecules. This energy is taken from the
leaf and transferred to the water molecules and released as gas molecules.
In the substomatal cavity, the water vapor pressure is in equilibrium with
the apoplast fluid facing the gas phase in the cavity. When water vaporizes
from the substomatal cavity, a new equilibrium establishes between the
water vapor pressure of the cavity and cells. Consequently, the water vapor
and associated energy are released into the atmosphere via the stomata
(thermal dissipation). Therefore, transpiration of water vapor from the sto-
mata is associated with cooling of the leaf (Hetherington and Woodward,
2003). For example, using infrared thermal imaging, it has been shown that
mutants with open stomata have lower leaf surface temperature than wild-
type plants (Merlot et al., 2002). Therefore, proper functioning of stomata
is vital for balancing leaf temperature and water loss. In Arabidopsis plants
that had developed at high temperature (28 ° C), increased water loss was
associated with an enhanced leaf cooling capacity. However, the leaves of
the high-temperature-developed plants possess lower stomatal density and
reduced stomatal size. In this case, to cool down the leaf, plant architec-
tural adaptions such as petiole elongation, leaf elevation above the soil, and
decrease in leaf thickness enhanced the diffusion of water vapor from the
stomata (Crawford et al., 2012; Murata and Mori, 2013).
As another role of stomata, it is generally accepted that the tran-
spirational flux is the main mechanism of transport of nutrients in plants
(Mengel and Kirkby, 1982; Novák and Vidovič, 2003).

6.2.2  Mechanism of Stomatal Closure and Opening

Closure and opening of the stomata pores depend on many factors, including
environmental factors such as light, temperature, and RH (or better water
VPD), CO2 concentration inside the stomatal cavity, water availability, and
pathogens, and endogenous factors such as phytohormones and their inter-
actions, and secondary messengers. Stomatal aperture changes over diur-
nal cycles. To facilitate CO2 assimilation, stomata stay open during the day,
especially in response to blue light, and tend to be closed at night (Talbott
and Zeiger, 1998; Schroeder et al., 2001; Tallman, 2004). However, to con-
serve water, plants with a crassulacean acid metabolism (CAM) close their
stomata during the daytime and open at night to uptake CO2 (Bohnert et al.,
1995; Black and Osmond, 2005). In this way, CAM plants lose less water and
are adapted to dry conditions. The number of stomata over the leaf surface
is species dependent (Table 6.1) and highly depends on the environmental

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Table 6.1  Stomatal Density on the Leaf of Some Horticultural Crops

Stomatal Density
Species (no. mm−2) Reference
Faba bean 30–50 Aliniaeifard et al. (2014)
Rose 37–65 Torre et al. (2003), Fanourakis et al.
(2011, 2013a), Giday et al. (2013a)
Tradescantia 19–22 Rezaei Nejad and van Meeteren (2005)
Cucumber 322–460 Bakker (1991), Savvides et al. (2012)
Tomato 75–203 Willmer and Johnston (1976), Bakker
(1991), Amjad et al. (2014)
Eggplant 136–182 Bakker (1991)
Arabidopsis 100–600 Doheny-Adams et al. (2012)
Anthurium 100–220 Schroeder and Stimart (2005)
Strawberry 120–330 Blanke (2002), Orsini et al. (2012)
Papaya 265–400 Parés et al. (2008)
Basil 50–250 Barbieri et al. (2012)
Almond 150–326 Camposeo et al. (2011)
Potato 80–260 Yan et al. (2012)
Gerbera 116–194 Romero-Aranda et al. (1994)
Azalea 108–163 Ceulemans et al. (1984)
Dodonaea 320–430 Shtein et al. (2011)
Apple 320–575 Blanke (1987), Blanke and Lenz (1989),
Blanke et al. (1994)
Trigonella 225–245 Willmer and Johnston (1976)
Orange 360–500 Moreshet and Green (1980), Burg
(2004)
Banana 1700 Wardlaw (1936)

condition prevalent during growth of the plants. The distribution of stomata

over the leaf surface is not a random process. They are distributed in a way
that effective diffusion of the gases between leaf internal tissues and the
surrounding environment is provided. Therefore, development of stomata
must be conserved in a good order to support their roles. According to the
one-cell spacing rule (Sachs, 1991), around the ­stomata is always at least
one nonstomatal epidermal cell that separates stomata from each other.
Since stomatal opening and closure require efficient trafficking of water
and ion molecules between guard cells and neighboring nonstomatal cells,
the one-cell spacing rule is vital for proper regulation of stomatal aperture
(Peterson et al., 2010). Stomatal opening and closing are controlled by guard

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 Stom ata a n d P ostharvest P hy s i o l o g y

cell swelling and shrinking, respectively. Guard cell turgor dynamically

changes in response to environmental conditions and endogenous signals.
Since guard cells lack plasmodesmata, most of the effluxes and influxes of
the solutes are accomplished through pumps, ion channels, and transport-
ers located in the plasma membrane (Daszkowska-Golec and Szarejko,
2013). Stomatal opening is initiated by extrusion of protons from guard cell
membranes through H+ -ATPases. Signals such as blue light and auxin are
positive regulators of H+ -ATPases, while calcium and the phytohormone
abscisic acid (ABA) operate as negative regulators of H+ -ATPases (Hentzen
et al., 1996; Parvathi and Raghavendra, 1997; Elzenga and Van Volkenburgh,
1997; Frechilla et al., 1999; Leonhardt et al., 1999; Elzenga et al., 2000; Zeiger,
2000; Zhang et al., 2004, 2007; Takemiya et al., 2006; Marten et al., 2007a;
Merlot et al., 2007; Su et al., 2007; Harada and Shimazaki, 2009; Zhao et al.,
2012). Proton extrusion induces plasma membrane hyperpolarization and
apoplast acidification. The created voltage gradient activates inward-recti-
fying K+ channels. Influx of K+, which is balanced by the influx of counter-
ions such as Cl− and NO3− and inorganic solutes such as malate, enhances
guard cells’ osmotic potential. Therefore, water pumped into the guard cells
causes swelling of the guard cells, and as a result, stomatal opening occurs.
Stomatal closing is initiated by inhibition of the H+ -ATPases, which depolar-
izes the plasma membrane. Following depolarization, outwardly rectifying
K+ channels enhance the driving force for K+  efflux and decrease the K+
level inside the guard cells. Two distinct types of depolarization-activated
anion channels act in the plasma membrane of the guard cells during sto-
matal closure: one activates rapidly by depolarization and also deactivates
rapidly by hyperpolarization (R-type anion channels), and the other one
activates and deactivates very slowly by voltage gradients (S-type anion
channels) (Schroeder and Keller, 1992). Anion channels facilitate efflux
of malate2−, Cl−, and NO3− from the guard cells. Moreover, gluconeogenic
conversion of malate2− into starch would result in further decrease of the
malate2− level. Cytosolic Ca 2+ elevation (also oscillation) through channels
located in the plasma membrane and in the tonoplast usually accompanies
stomatal closure (Leckie et al., 1998; Siegel et al., 2009; Hubbard et al., 2012;
Daszkowska-Golec and Szarejko, 2013).
Efflux of K + and Cl− ions or malate2− through guard cells’ membrane
decreases osmotic potential, and as a result, water exits from the guard
cells, which causes them to shrink, resulting in stomatal closure (Blatt,
2000; Schroeder et al., 2001; Outlaw, 2003).
Plants dynamically respond to changes in environmental conditions
by regulating the aperture of the stomata. Plant responses to environmen-
tal stresses (e.g., drought) are usually associated with induction of abscisic
acid (ABA) production. ABA, through its signal transduction pathway,
causes stomatal closure (Hu et al., 2006; Endo et al., 2008; Lee and Luan,
2012; Sreenivasulu et al., 2012).

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6.2.3  Signal Transduction Pathways

in Guard Cells for Stomatal Closure
Guard cells perceive multiple signals from the environment and integrate
them to internal signals. By following complex transduction pathways, guard
cells respond by regulating stomatal aperture in order to adapt to the envi-
ronment (Bohnert et al., 1995; Qin and Zeevaart, 2002; Lebaudy et al., 2008;
Oh et al., 2009; Kim et al., 2010; Trontin et al., 2011; Lee and Luan, 2012; Zhu
et al., 2012; Christmann et al., 2013; Kuromori et al., 2014). Over the past
several years, many internal signals have been recognized in guard cells in
response to different environmental signals. For example, calcium, reactive
oxygen species (ROS), phosphatidic acid, cyclic guanosine 3',5’-monophos-
phate (cGMP), nitric oxide (NO), and pH have been found as essential sig-
nals mediated in stomatal closure (Suhita et al., 2004; Li et al., 2006; Wang
and Song, 2008; Xue et al., 2009; Dubovskaya et al., 2011; Kim et al., 2010;
Stael et al., 2011). Although an ABA-independent pathway for closure of the
stomata has also been proposed (Yoshida et al., 2006; Huang et al., 2009;
Montillet et al., 2013; Roychoudhury et al., 2013), ABA is considered to be the
main phytohormone that promotes stomatal closure, which helps to mini-
mize water loss by decreasing transpiration via stomata. This function of
ABA is accomplished through modulating a complex and sophisticated cas-
cade of biochemical and molecular events (Hauser et al., 2011). Almost all of
the previously mentioned internal signals are involved in guard cells’ ABA
signaling pathway for closure of the stomata (Leung et al., 1997; Kwak et al.,
2002; Suhita et al., 2004; Li et al., 2006; Zhu et al., 2007; Neill et al., 2008; Wang
and Song, 2008; Hubbard et al., 2010, 2012; Kim et al., 2010; Dubovskaya
et al., 2011; Joshi-Saha et al., 2011; Hossain et al., 2011). The ABA-induced
stomatal closure is often associated with an increase in guard cells’ calcium
concentration. However, calcium-dependent and ­calcium-independent ABA
signaling pathways have been suggested for ABA-induced stomatal closure
(Figure 6.2) (MacRobbie, 1990; Li and Assmann, 1996; Levchenko et al.,
2005; Sutter et al., 2007; Marten et al., 2007b; Geiger et al., 2009; Siegel et al.,
2009; Geiger et al., 2010; Joshi-Saha et al., 2011).
To initiate ABA signal transduction, guard cells are equipped with ABA
receptors to bind to ABA (Moes et al., 2008; Fujita et al., 2009; Ma et  al.,
2009; Santiago et al., 2009; Cutler et al., 2010; Raghavendra et al., 2010; Lee
et al., 2013). Binding of ABA with its receptors inhibits the activity of group
A protein phosphatases 2C (PP2Cs) (Moes et al., 2008; Fujita et al., 2009; Ma
et al., 2009; Park et al., 2009; Santiago et al., 2009; Raghavendra et al., 2010).
OST1 is an Arabidopsis SnRK2-type protein kinase that, together
with several other SnRK2-type protein kinases, is also known to func-
tion in ABA responses (Belin et al., 2006; Fujita et al., 2009; Lee et al.,
2013; Yoshida et al., 2002, 2006). In contrast to A-type PP2Cs, SnRK2-type

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 Stom ata a n d P ostharvest P hy s i o l o g y

protein kinases are positive regulators of ABA signaling (Fujii et al., 2009;
Fujii and Zhu, 2009; Ma et al., 2009; Park et al., 2009; Umezawa et al., 2009;
Vlad et  al., 2009; Lee and Luan, 2012). Downstream of ABA receptors,
PP2Cs, and SnRKs are ion channels that control stomatal movements (Fujii
et al., 2009; Geiger et al., 2009; Lee et al., 2009). The guard cell slow-type
anion channel (SLAC1) may represent an essential component for stomatal
closure induced by ABA or other signals (Negi et al., 2008; Vahisalu et al.,

ABA

ABA Receptors
Calcium-independent ABA signaling

Calcium-dependent ABA signaling

PP2C

OST1 CDPKs

Ion channels
(e.g., SLAC1)

Stomatal
closure

Figure 6.2  Simplified ABA signal transduction pathway in guard cells for
closure of the stomata. In a calcium-independent ABA signaling pathway,
perception of ABA by receptors leads to inactivation of type 2C protein
phosphatases (PP2C). As a result, S-type anion channels (SLAC1) will be
activated by a SnRK2-type protein kinase (OST1). Consequently, stomatal
closure occurs. In the calcium-dependent ABA signaling pathway, calcium-
dependent protein kinases (CDPKs) can induce stomatal closure via activa-
tion of SLAC1 (Data from Hubbard, K.E. et al., Genes and Development
24, 1695–1708, 2010; Kim, T.H. et al., Annual Review of Plant Biology
61, 561–591, 2010; Antoni, R. et al., Current Opinion in Plant Biology 14,
547–553, 2011; Sreenivasulu, N. et al., Gene 506, 265–273, 2012; Lee,
S.C. et al., Molecular Plant 6, 528–538, 2013).

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2008, 2010; Geiger et al., 2009). SLAC1 acts as a substrate for and is acti-
vated by OST1 (Geiger et al., 2009; Lee et al., 2009, 2013).
Calcium-dependent protein kinases (CDPKs) function as essential
elements of the calcium-dependent ABA signaling (Zhu et al., 2007). It has
been shown that SLAC1 can be activated by CDPKs, which leads to stoma-
tal closure (Mori et al., 2006; Geiger et al., 2010) (Figure 6.2).

6.3  Role of Stomata in the Postharvest Phase

As stomata play an important role in controlling gas exchange between
internal plant tissues and the surrounding environment, they can affect the
postharvest physiology of plants and plant products as far as postharvest
physiology is influenced by gas exchange. In this chapter, we discuss spe-
cific postharvest issues of cut flowers, vegetables, and fruits that are related
to the behavior of stomata in these products.

6.3.1  Stomata in Relation to Vase Life of Cut Flowers

The vase life of cut flowers ends when the flowers have lost their ornamen-
tal value. Most flowers attached to the plant lose their ornamental value
because of senescence symptoms as a result of their advanced development;
they show changes in color, wilting of petals, closing of flowers, or abscis-
sion of flowers or petals. However, when flower shoots are cut and placed
in water, they often lose their attractiveness for the consumer because of
premature wilting of flowers or leaves, or loss of stem turgidity; these are
all symptoms of a water deficit (Van Doorn, 1997, 2012). This water deficit
is a result of a negative water balance: the rate of water uptake has become
less than the rate of water loss by transpiration. Other consequences of the
negative water balance for the quality of cut flowers can be impairment of
flower opening and precocious senescence.
In most cases, the imbalance between uptake and loss of water is
the result of a hampered water uptake, due to a gradual decrease in the
hydraulic conductance of the cut flower stem after cutting (Aarts, 1957; van
Meeteren, 1978, 1989; Rogers et al., 1979; Halevy and Mayak, 1981). There
can be several reasons for this decrease in stem hydraulic conductance:
growth of bacteria at the cut stem surface or inside the xylem vessels,
uptake of air into the cut-open xylem vessels at the cut stem surface (air
embolism), wound-induced responses in the cut stem, or formation of vapor
cavities in the xylem fluid (cavitations) (Tyree and Dixon, 1986; Williamson
and Milburn, 1995; Van Doorn, 1997). Because of the ongoing transpira-
tion of the cut flower, the decrease in hydraulic conductance of the stem
results in a decrease of the water potential of the flower and loss of turgidity.

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The decrease in water potential can cause cavitations, which will further
enlarge the imbalance between water uptake and loss.
The negative effect of a decreased stem hydraulic conductance on the
water balance of a cut flower will be postponed or less severe when the tran-
spiration rate of the cut flower is low. As an example, it has been shown that
the vase life of cut roses can be prolonged by various ways that lower their
transpiration: cooling, high air humidity, and leaf removal (Zieslin, 1989).
Mayak and Halevy (1974) showed that the difference in vase life between a
short-lived cultivar and two long-lived cultivars of rose was large, especially
under conditions promoting high transpiration rates, and was narrowed when
either flowers were held in mild conditions or the leaves were stripped off.
Most cut flowers include, besides one or more flowers, a stem that
often bears leaves. The main parts of a flower consist of a vegetative part,
containing sepals and petals, and a reproductive part, containing stamens
and a gynoecium. Because a waxy cuticle covers the outer surface of the
epidermal cells of leaves, sepals, petals, and the gynoecium, most of the
transpiration of cut flowers is via the stomata. Therefore, it can be expected
that the number and the closing and opening behavior of stomata play a role
in vase life. Stomata are usually present in green epidermal tissues such as
leaves, sepals, and green (parts of) petals (Chimona et al., 2012), but also in
some nongreen petals, stomata can be found (Effmert et al., 2005; Chimona
et al., 2012). Stomata can also be completely absent in petals (Mayak and
Halevy, 1974; Tahir and Rajput, 2010). In petals of commercially important
crops like roses (Rosa hybrida), carnations (Dianthus caryophyllus), and ger-
bera (Gerbera jamesonii), stomata were not found (Van Doorn, 1997). As
shown in the overview by Van Doorn (1997), numbers of stomata in petals
vary between 0 and 45 per cm 2, which is very low compared with the num-
bers of stomata found in leaves, which varies between 5 and 1000 per mm 2,
depending on species and growth conditions (Table 6.1) (Hetherington and
Woodward, 2003). When present, stomata in petals are not always func-
tional, as shown for several orchids (Hew et al., 1980). An exceptional flower
in this context is tulip; tulip petals can contain 5–49 stomata per mm 2, and
a temperature-dependent stomatal movement in tulip petals controls water
transpiration during flower opening and closing (Azad et al., 2007). In con-
clusion, for most cut flowers, the main transpiration is via the stomata in the
leaves of the cut flowering stem.
The role of stomata in vase life was elegantly demonstrated by Halevy
et al. (1974) in an experiment about the effects of application of ABA on
the vase life of cut roses. Application of ABA can induce stomatal closure,
reduce water loss, and extend the vase life of cut rose flower shoots bear-
ing leaves. Its effect was very pronounced when the flowers were exposed
to conditions that promote transpiration (28°C and 45% RH). However, in
a stomata-less system (leafless flower shoots) or in leafy shoots held in
darkness when all stomata were closed, ABA shortened vase life because it

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increased the aging of the flowers and some biochemical processes associ-
ated with it (RNase activity and reduction in protein content).
Elibox and Umaharan (2008) evaluated the vase life of 26 anthurium
(Anthurium andraeanum Hort.) cultivars. Among the tested cultivars, there
were large differences in vase life, varying from 14 to 49 days. Anthurium cut
flowers do not hold leaves on their cut stem and have a low transpiration rate
and long vase life. Still, symptoms associated with the end of the vase life in
anthurium are typical of water stress (Paull, 1982). Anthurium cut flowers
have a brightly colored modified leaf, the spathe, which subtends a spadix
with more than 300 spirally arranged minute flowers. This spathe contains
stomata, mostly on the abaxial side, of which the density varies between
the cultivars from 1.8 to 25.7 per mm 2. Although the rate of water loss of
anthurium cut flowers is relatively low, of 12 morphophysiological character-
istics, only spathe color and abaxial stomatal density were able to accurately
predict vase life (Elibox and Umaharan, 2008). In their study, Elibox and
Umaharan (2008) also showed that changes in vase life of cultivars over
season were mediated through changes in stomatal density. However, culti-
var differences in stomata density of the spathe explained only a small pro-
portion of cultivar variation in vase life. In a later study, the same authors
showed that the balance between factors affecting water uptake, which is
determined by the timing, extent, and duration of vascular occlusion, and
those affecting water loss (e.g., stomatal density and regulation) may con-
tribute to cut flower senescence in anthurium (Elibox and Umaharan, 2010).
In an evaluation of snapdragon (Antirrhinum majus) cultivars, that
differed in length of vase life, leaf stomatal numbers and postharvest water
loss were shown to be important factors in vase life. Cut flowers of a geno-
type with 9 days longer vase life had 53% fewer leaf stomata (Schroeder
and Stimart, 2005). However, long vase life was especially associated with
an early reduction in transpiration followed by low, steady transpiration,
while short-lived genotypes had a linear transpiration pattern over the vase
period. This indicates that differences in stomatal control (opening and
closing) of water loss between cultivars are likely to be a more important
factor than the number of stomata (Schroeder and Stimart, 2005). This
agrees with a previous paper of Rutland et al. (1987), in which they showed
that stomatal density did not correlate with vase life and transpiration of 10
different snapdragon cultivars.
Number of stomata (stomatal density) is not only dependent on geno-
type (cultivar), but also affected by the environmental conditions during
the cultivation of the cut flowers. Four of five rose cultivars tested showed
a significantly higher number of stomata per cut flower at higher cultiva-
tion temperatures (Pandey et al., 2007). However, the vase life of the cut
flowers was not affected by the preharvest temperature conditions (Pandey
et al., 2010). This strengthens the conclusion that stomata number per se
is not the determining factor for vase life. High air humidity (RH) during

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cultivation greatly reduced the vase life of cut roses, which was accompa-
nied by high transpiration rates (Torre, 1999; Mortensen and Gislerod,
2000) and symptoms that are typically related to water stress, including
premature flower and leaf wilting, as well as pedicel bending (Mortensen
and Gislerod, 2000; Torre and Fjeld, 2001; In et al., 2007). Roses grown at
high RH (90–95%) showed a significantly higher number of stomata and
longer stomata, as well as wider stomatal apertures, than roses grown at
a moderate RH of 60%–70% (Torre et al., 2003; Fanourakis et al., 2013a).
However, in rose leaflets subjected to desiccation, the enhanced stomatal
conductance in high-RH- as opposed to moderate-RH-grown plants was
mostly due to poor stomatal closure and, to a lesser extent, the combined
result of higher stomatal density and longer pore length (Fanourakis et al.,
2013a). This confirms that the reduced degree and, especially, the reduced
rate of stomatal closure are the primary causes of the large genotypic varia-
tion in the control of water loss in high-RH-grown rose plants. It is known
that stomata respond rapidly to air humidity, resulting in higher stomatal
conductance at high RH (Hall et al., 1975; Morison and Gifford, 1983).
Growth of rose plants at high RH, however, resulted in stomata that are
less responsive to desiccation. As a result, transpiration of the cut flowers
remains also high when the water potential in the cut flower decreases. This
implies that cultivation of cut flowers at high RH makes the flowers more
vulnerable to dry storage or transport, and to a hampered water uptake at
the consumer’s, caused by a decreased hydraulic stem conductance of the
cut flower. A limited exposure period to high RH of a few days can already
disturb the stomata closure response to desiccation (Rezaei Nejad and van
Meeteren, 2008; van Meeteren et al., 2009; Aliniaeifard et al., 2014). This
suggests that a period of some days of high RH during the preharvest stage
can have a large impact on the postharvest quality of cut flowers.

6.3.2  Stomata in Relation to Quality of Vegetables

In this chapter, we use the nonbiological definition of vegetables, which is
based on culinary and cultural traditions. In culinary terms, a vegetable is
an edible plant or its part, intended for cooking or eating raw (Vainio and
Bianchini, 2003). In botanical terms, a vegetable can be a root, stem, leaf,
immature flower bud, fruit, or sprout. Fruit vegetables often have a savory
flavor, while culinary fruits (discussed in the next part) often have a sweet
taste and are used for desserts or as a snack. As discussed before, stomata
can be found in most green plant parts. That implies that root-type vegeta-
bles, such as carrot, sweet potato, beet, and radish, are vegetables without
stomata. As the majority of stomata are on the leaves of plants, leafy veg-
etables will have a large number of stomata. But we can also find stomata in
many fruits, especially in green fruits, as most immature fruits are. Fruits of

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cucumber, okra, and several legumes, such as pea and bean, are used in an
immature stage as a vegetable. The stomata number on pickle cucumbers
varies from 10 to 100 mm−2 (Smith et al., 1979), what is comparable to some
leaves (see before). The stomatal density decreases when the cucumber
fruit size increases, because stomata are differentiated in an early stage of
fruit development. So, the total number of stomata per fruit does not change
during growth of the fruit. In the legume plants, pots are also rich in sto-
mata. In the pot of broad bean or pea, stomata in the exocarp are 25% of the
stomatal density on the leaf abaxial surface and 50% of the stomatal density
on the adaxial side of the leaf (Willmer and Johnston, 1976; Atkins et al.,
1977; Blanke and Lenz, 1989). The stomatal density in broad bean is around
19 stomata mm−2, and in pea is around 40 stomata mm−2. It has been shown
that 12,000–14,000 stomata exist on one pea pod. The maximum density of
stomata in the pea can be observed after anthesis (Blanke and Lenz, 1989).
Water loss is one of the most obvious changes in harvested ­vegetables,
often being the factor that limits marketing life (Ben-Yehoshua and Rodov,
2003; Shamaila, 2005). Therefore, it is surprising that the effects of prehar-
vest growth conditions and postharvest handling practices on the stoma-
tal aperture of harvested vegetables have received little attention. Water
loss during storage can be controlled with proper and rapid precooling and
storage humidification. In chilling-sensitive vegetables, however, control of
water loss is more difficult, since they can only be stored at warmer tem-
peratures. Also, the environmental conditions in different links of a supply
chain are not always under strict control, which makes the role of stomata
of even more importance. In a study by Thomson (2005), observations
were made on stomatal apertures in the leaves of Japanese mustard spin-
ach (Brassica rapa var. perviridis ‘Komatsuna’) and a large India mustard
(Brassica juncea ‘Red Giant’) in response to cooling and holding treatments
representing commercial procedures. When the leaves were held in air at
20°C, the stomatal aperture of both Brassicas progressively reduced after
harvest; after 15 min, closure was largely complete in B. juncea, while in
B. rapa, stomata closure took about 30 min. Stomata of B. juncea also closed
at 4°C; however, 4°C inhibited stomatal closure of B. rapa. Besides cool-
ing by air, following harvest, B. rapa leaves were also dipped for 20 min in
iced water and in water at 20°C. Neither water dips interfered with stoma-
tal ­closure. However, after these dips and a further 25 min in air at 20°C,
stomata remained largely closed for iced leaves, but had begun to open on
leaves dipped in the 20°C water.
Reducing water loss in B. juncea not only prevented wilting, but
also reduced leaf yellowing; it increased the sweetness and retarded pro-
tein degradation and the loss of vitamin C (Lazan et al., 1987a, 1987b).
Also, in lettuce, the retention of water played an important role in main-
taining ascorbic acid content (Agüero et al., 2011). In general, the nutri-
tional value of vegetables is very much affected by water loss (Paull, 1999;

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Ben-Yehoshua and Rodov, 2003; Shamaila, 2005). However, there has only
been limited research evaluating the impact of water loss on the nutritive
value of vegetables. Ezell and Wilcox (1959, 1962) found that water loss was
associated with significant losses of both vitamin C content and carotene in
a wide range of vegetables (kale, collards, turnip greens, spinach, cabbage,
and snap beans). Diminishing water loss of broccoli by misting resulted in
significant retention of ascorbic acid (Barth et al., 1992). Also, preventing
water loss of strawberries (by wrapping) reduced the loss of ascorbic acid.
As modifications of O2 and CO2 levels were only minimal in the wrapped
treatments, the effect was not due to their modification (Nunes et al., 1998).
Water loss is clearly an important factor in the retention of quality, includ-
ing nutritional quality, in vegetables. As discussed in Section 6.4, the culti-
vation of plants at low VPD can influence the closing ability of stomata. In
that way, the VPD during the preharvest phase of vegetables influences the
postharvest life and nutritional quality of leafy vegetables via the stomata. A
higher rate of water loss after harvest of plant leaves was observed in basil
(Ocimum basilicum L.) and lemon balm (Melissa officinalis L.) as a result
of exposure during cultivation to low VPD than in plants grown at higher
VPDs (Islam et al., 2010). Postharvest life was negatively correlated to the
rate of water loss via stomata after harvest of the plants (Islam et al., 2010).
Whether VPD during the growth of legume plants also affects the stomatal
closing ability of the pots is unknown.
Some recent outbreaks of bacteria-related diseases after consumption
of fresh vegetables have increased the interest in food-borne pathogens and
especially in bacteria–plant interactions. Since stomata are the portals into
the plant for many bacterial species that cause plant diseases, there is more
and more interest in the role that stomata play in food safety, especially
in leafy greens. Scanning electron microscopy studies of lettuce leaves
showed that bacteria can infiltrate stomata and in that way are protected
from chemicals such as sodium hypochlorite during the washing of the
leaves (López-Gálvez et al., 2010). Therefore, washing can be ineffective in
eliminating Escherichia coli cells on leaf surfaces, and such is also the case
for sanitizers used in the washing solution. The main function of sanitizers
in the washing of fresh-cut products is to avoid cross-contamination.
Incubation of GFP-tagged Salmonella enterica with iceberg lettuce
leaves in the light resulted in aggregation of bacteria near open stomata
and invasion into the inner leaf tissue (Kroupitski et al., 2009). In contrast,
when stomata were closed due to darkness, incubation resulted in a scat-
tered attachment pattern and very poor stomatal internalization. However,
Salmonella internalization was not enhanced by forcing stomatal opening
in the dark by the use of fusicoccin. These results imply not only that light
was necessary for stomata opening, but also that the pathogen is attracted
to nutrients produced de novo by photosynthetically active cells. Some plant
pathogens force stomata to open after they have closed as part of the plant’s

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initial response to those invasive bacteria. However, lettuce stomata do not

close in response to Salmonella, probably because this bacterium is not a
plant pathogen (Kroupitski et al., 2009). Also, a few listeriosis outbreaks
have been linked to consumption of contaminated vegetables. By dipping
plants of Arabidopsis thaliana for 5 min into water containing Listeria mono-
cytogenes expressing GFP and rinsing repeatedly, it was demonstrated that
these bacteria can internalize the plants via their stomata (Milillo et  al.,
2008). The bacteria were visualized in the intercellular spaces of the leaves
and recovered from the leaves at densities from 1.52 to 2.17 log CFU cm−2 .
Ten days after exposure, bacterial numbers had increased over initial
numbers by 2.60–2.95  log  CFU cm−2 . Taking measures to limit the pen-
etration process of bacteria in leafy vegetables via stomata could improve
food safety.

6.3.3  Stomata in Relation to Quality of Fruits

It is generally accepted that the main plant structure responsible for gas
exchange between a plant and its environment, stomata, is distributed over
the leaf surface. However, some structures similar to the leaf stomata have
been found in the fruits of many plants. Increased fruit transpiration at
light and decreased transpiration at darkness, together with the morphol-
ogy of a fruit’s guard cells, confirm that fruit’s stomata can function like
those on the leaves (Blanke and Lenz, 1989). Fruits of some horticultural
crops possess stomata, while others lack them. The number of stomata
over the fruit surface is species dependent (Table 6.2) and depends highly
on the stage of the fruit development and the environment. For some crops,
different results regarding the existence of stomata have been reported.
For example, the presence of stomata on pepper fruit has been reported
by Fallik et  al. (1999), while Smith et al. (2006) reported no stomata on
pepper fruit. In some fruits, the stomata are not evenly distributed over
the fruit surface. For instance, the stylar scar region of sweet cherry has
the highest stomatal density, followed by the cheek, while the stem cavity
region is lacking stomata (Peschel et al., 2003). Stomata in sweet cherry
are directly or indirectly involved in water transport. When stomata are
functional, their role in water transport varies during the course of a day,
and they may be relevant to the process of fruit cracking (Christensen,
1976; Peschel et al., 2003).
In some crops at early stages of fruit development, stomata exist on
the fruits (especially when they are green), while with progress toward rip-
ening their number decrease or sometimes they disappear at the late stage
of fruit development (Blanke and Lenz, 1989; Rogiers et al., 2004). Grape
berries contain some functional stomata at the preveraison stage, but they
become clogged with wax and nonfunctional in the postveraison stage of

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Table 6.2  Stomatal Density on the Fruits of Some Horticultural Crops

Species Stomatal Density Reference
Strawberry 1 – 6 (no. mm )
−2 Blanke (2002)
Cucumber 20 – 30 (no. mm−2) Adams and Ho (1995)
Tomato –1 to 0 (no. mm−2) Willmer and Johnston (1976),
Adams and Ho (1995)
Grape 8 – 10 (per berry) Blanke and Leyhe (1988)
Sweet cherry 143 – 2,124 (per fruit) Peschel et al. (2003)
Apple 600 – 6,000 (per fruit) Blanke and Lenz (1989); Blanke
1 (no. mm−2) (1994)
Faba bean 19 ± 2 (no. mm−2 on pod) Willmer and Johnston (1976),
Blanke and Lenz (1989)
Pea 40 (no. mm−2 on pod) Blanke and Lenz (1989)
Orange 6 – 75 (no. mm−2) Turrell and Klotz (1940),
Moreshet and Green (1980),
Blanke (1994)
Banana 3.2 – 11 (no. mm−2) Wardlaw (1936), Johnson and
Brun (1966)
Avocado 20,000–30,000 (per fruit) Blanke (1992, 1994)
3 – 5 (no. mm−2)
Trigonella 61 ± 2 (no. mm−2 on pod) Willmer and Johnston (1976)

fruit development (Rogiers et al., 2004). The transformation from functional

stomata at the preveraison stage to nonfunctional stomata (lenticel covered
by wax) at the postveraison stage causes a decrease in berry transpirational
water loss at the late stage of fruit ripening (Rogiers et al., 2004). Therefore,
the flesh of the grape berry at the postveraison stage is highly isolated from
the outside environment, causing higher CO2 and lower O2 concentrations
in the berry core than in fruits with a high number of stomata, such as pear
(Rogiers et al., 2004; Franck et al., 2007; Mencarelli and Bellincontro, 2013).
The berries are attached to the stem through a pedicel. The pedicel is cov-
ered with a thin layer of cuticle and contains a high number of stomata and
lenticels. These properties make the pedicel vulnerable to water loss when
the RH of the surrounding environment is low. High permeability of the
pedicel and its high water loss properties induce a water potential difference
that draws water from the berry, leading to water loss from the airtight ber-
ries (Mencarelli and Bellincontro, 2013).
Young apple fruits are covered by a thin cuticle after petal fall. In this
stage, stomata are completely formed and uniformly distributed over the
fruit (Schwertfeger and Buchloh, 1968). The number of stomata in apple
fruit is determined before petal fall. Stomatal density in apples is reported

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between 10 and 20 stomata mm−2 around the petal fall (Blanke, 1986; Blanke
and Lenz, 1989). Then after 1 week, it decreases to 6–8 stomata mm−2 , and
thereafter, when the expansion of the cuticle occurs, it decreases further to
less than 1 mm−2 (Schwertfeger and Buchloh, 1968; Blanke, 1986; Blanke
and Lenz, 1989). It is reported that an apple fruit with 10 mm diameter
would have around 600–6000 stomata per fruit (Table 6.2) (Blanke and
Lenz, 1989).
Also in Valencia oranges, the stomatal density is at its maximum
(160  mm−2) when the fruit is small. Fruit growth expands the space
between the stomata; in large fruits, the number of stomata is decreased
to 20–50 mm−2 (Moreshet and Green, 1980). In navel oranges, the stoma-
tal density (10–16 mm−2) is lower than that in the fruit of Valencia orange
(Turrell and Klotz, 1940). It seems that in all fruits, even when they are ripe,
stomata exist on them, but they are covered by a wax layer.
In banana fruit, contrary to the leaf, the stomata are not distributed in
a parallel way. The number of stomata is between 3 and 11 mm−2 in imma-
ture fruits and 3 and 6 mm−2 in full and mature fruits (Johnson and Brun,
1966). The stomatal density in the leaf (Table 6.1) of banana (1700 mm−2) is
considerably higher than their numbers on the fruits (Table 6.2) (Wardlaw,
1936). Although many studies reported nonfunctional stomata in banana
fruit, it has been shown that just before the harvest, they are functional in
response to light and CO2 and capable of gas exchange with the environ-
ment (Moreshet and Green, 1980; Burg, 2004).
Stomatal density in tomato fruits is very low, and in some studies,
even no stomata were reported on the tomato fruit, while the number of sto-
mata in cucumber is relatively high (Table 6.2). Higher stomatal densities
correlate positively with higher water loss from the fruits. Measuring the
amount of water loss immediately after harvest showed a 20 times higher
water loss for cucumber fruits than for tomato fruits (Adams and Ho, 1995).
In ABA-deficient orange plants, an increase in stomatal opening on the fruit
caused higher fruit water loss than in wild-type oranges (Romero et al.,
2013). This also demonstrated the role of stomata in the water loss of fruits.
Therefore, fast handling after harvest is important in order to restrict water
loss and quality loss in fruits with higher stomatal densities.

6.4  Preharvest Conditions Leading to

Postharvest Problems via Stomata
There are several preharvest factors that can influence the postharvest per-
formance of horticultural crops (Sams, 1999; Lee and Kader, 2000; Herppich
et al., 2001; Torre and Fjeld, 2001; Tijskens et al., 2003; Rezaei Nejad and
van Meeteren, 2005; Hewett, 2006; In et al., 2007; Fonseca et al.,  2009;

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Moran  et  al., 2009; Islam et al., 2010; Fanourakis et al., 2011; Burchi and
Prisa, 2013; Tudela et al., 2013). During growth of the plants, the manage-
ment of environmental conditions such as relative humidity (RH), light, and
temperature can considerably influence postharvest performance of horti-
cultural products. Some of these conditions affect postharvest performance
via effects on the stomata (number, morphology, and closing ability).

6.4.1  Relative Humidity and Stomata Control

Commercial production of vegetables, cut flowers, and ornamental plants
under closed and semiclosed environments has tremendously increased
over the last few decades. In this kind of protected cultivation system,
often the RH inside the production facility is high because of energy sav-
ing (reduced ventilation and increased insulation) and transpiration by the
plants. RH is the ratio of the partial pressure of water vapor of the air to the
saturated water vapor pressure, expressed in percentages. Air saturates
when it holds water with its maximum capacity, which depends on tem-
perature. More moisture beyond this capacity would lead to condensation
of water vapor molecules to liquid water. By increasing the temperature,
the maximum water holding capacity of the air increases. Therefore, the
RH depends on the actual water vapor pressure of the air and air tempera-
ture. Vapor pressure deficit (VPD) is defined as the difference between
the saturation water vapor pressure and the actual water vapor pressure.
According to Fick’s first law, the diffusion rate of water vapor from a leaf
is linearly related to the difference in water vapor pressure inside and out-
side the leaf. Therefore, the humidity in plant environments is preferably
expressed as VPD.
Plants that have been produced under low-VPD conditions (very
humid) have often grown normally, but after harvest their stomata do not
function in a normal way anymore. Due to the prolonged exposure to low
VPD, a habituation process occurs in guard cells of the stomata, which
makes the stomata insensitive to stimuli that would normally provoke sto-
matal closure (stomatal malfunctioning) (Rezaei Nejad and van Meeteren,
2005; Aliniaeifard et al., 2014). As a result, stomata also stay open when
they encounter desiccation after harvest of the plants or plant products.
This disturbance in the normal functioning of the stomata due to previ-
ous exposure to low VPD therefore has horticultural consequences. For
example, as discussed before, growing rose plants at low-VPD conditions
often results in a decrease in vase life after harvest of the plants (Torre
et al., 2001). Low-VPD-grown plants often have a higher rate of water loss
than the moderate-VPD-grown plants; this is also the case during desic-
cation or hampered water uptake (postharvest stage) or when they are

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exposed to high VPDs (Rezaei Nejad and van Meeteren, 2005; Rezaei
Nejad et al., 2006; Aliniaeifard and van Meeteren, 2013; Fanourakis et al.,
2013a; Aliniaeifard, 2014; Aliniaeifard et al., 2014). Such a transfer from
low to high VPD is common after vegetative propagation by leafy cuttings,
after in vitro propagation, and at the end of the cultivation period of orna-
mentals when plants or cut flowers are transferred to domestic conditions.
Although plants can be cultured in vitro on a large scale under low-VPD
conditions, in vitro–produced plants are usually susceptible to wilting
upon transfer to normal atmospheric VPDs (Brainerd and Fuchigami,
1982; Ghashghaie et al., 1992; Santamaria et al., 1993; Aguilar et al., 2000;
Hronková et al., 2003; Hazarika, 2006; Aracama et al., 2008; Khan et al.,
2010). This is also because of malfunctioning of the stomata in response to
a wide range of closing stimuli, such as darkness, abscisic acid (ABA), and
elevated calcium (Ca 2+) levels (Brainerd and Fuchigami, 1982; Ziv et al.,
1987; Santamaria et al., 1993). It has been shown that higher rates of water
loss after desiccation (postharvest phase) in the plants grown at low-VPD
conditions are mainly caused by the stomata, compared to the role of the
cuticle (Ziv et al., 1987; Santamaria and Kerstiens, 1994; Aliniaeifard and
van Meeteren, 2013; Fanourakis et al., 2013a).
As discussed in specific sections before, cultivation at low VPD not
only influences the vase life and visual appearance of cut flowers, but also
can influence the postharvest life and nutritional quality of leafy vegetables
via the stomata. Leaves of plants that lose their moisture more easily are
more vulnerable to losing vitamin than plants that are resistant to wilting
(Ezell and Wilcox, 1959; Lee and Kader, 2000).

6.4.1.1  How Does Preharvest Low VPD Affect

Postharvest Stomata Control?
Stomata react very fast to changes in the environmental conditions through
internal signals (Martin and Meidner, 1971; Wigger et al., 2002; Tallman,
2004; Neill et al., 2008; Kim et al., 2010; Hao et al., 2011; Hossain et al.,
2011). In natural conditions, plants continuously encounter changes in the
environment. Therefore, in order to survive, they should have the ability
to react fast to the changing environment. ABA is the main phytohormone
that acts as an internal signal in response to changes in environmental
conditions and triggers changes in various plant physiological and develop-
mental processes, resulting in adaptation to the stress conditions, including
stomatal closure (Aguilar et al., 2000; Chen et al., 2010; Lee and Luan, 2012).
In general, the ABA level is regulated by a balance between its biosynthesis
and its catabolism (Nambara and Marion-Poll, 2005; Lee and Luan, 2012).
The first step in the specific ABA production pathway is the synthesis of
violaxanthin through zeaxanthine epoxidase. Neoxanthin synthase and an
isomerase may be required for formation of cis-isomers of violaxanthin

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and neoxanthin. 9-cis-Epoxycarotenoid dioxygenases (NCEDs) cleave the

cis-xanthophylls to yield xanthoxin. It has been shown that NCED is the
rate-limiting factor in the ABA biosynthetic pathway (Qin and Zeevaart,
1999; Iuchi et al., 2001; Tan et al., 2003). Through a short-chain alcohol
dehydrogenase (ABA2), xanthoxin is then converted to abscisic aldehyde.
Finally, abscisic aldehyde oxidase (A AO3) catalyzes the last step of ABA
biosynthesis by oxidation of abscisic aldehyde into ABA (Nambara and
Marion-Poll, 2005).
The place in the plant of ABA biosynthesis and the place of its action
are still under debate (Christmann et al., 2005; Davies et al., 2005; Endo
et al., 2008; Jiang and Hartung, 2008; Melhorn et al., 2008). Following
drought stress, root-produced ABA acts as a signal between roots and
shoots (Holbrook et al., 2002; Wilkinson and Davies, 2002; Davies et al.,
2005; Jiang and Hartung, 2008). However, it has also been indicated that
synthesis of ABA in the shoot is a response to a long-distance hydraulic sig-
nal in xylem vessels due to low water potential in the soil (Holbrook et al.,
2002; Christmann et al., 2005, 2007, 2013). Nonetheless, it has been shown
that NCED3 is mainly expressed and localized in the vascular parenchyma
of leaves (Cheng et al., 2002; Endo et al., 2008). The expression of AAO3 in
the guard cells has also been reported (Koiwai et al., 2004; Nambara and
Marion-Poll, 2005; Melhorn et al., 2008). Moreover, the transient expres-
sion of NCED3 or AAO3 in guard cells promotes stomatal closure (Melhorn
et al., 2008). Bauer et al. (2012) showed that guard cells possess the entire
ABA biosynthetic pathway. Guard cells are able to autonomously synthesize
ABA, and there is a positive feedback loop for ABA production when they
have been exposed to high VPDs around the leaves (Bauer et al., 2012). It
has been suggested that there is no need for root–shoot transport of ABA
(Osakabe et al., 2013). Moreover, increasing VPD around the leaves of well-
watered plants resulted in a higher ABA level in the leaf (Rezaei Nejad and
van Meeteren, 2007, 2008).
ABA inactivates mainly through oxidation or conjugation processes.
Hydroxylation of ABA is the main process for ABA inactivation (Nambara
and Marion-Poll, 2005). Oxidation of ABA is catalyzed by 8’-hydroxylases
to form 8’-hydroxy ABA. In the next step, 8-hydroxy ABA spontaneously
isomerizes to phaseic acid (PA) and is further reduced to dihydrophaseic
acid (DPA) through an unknown reductase (Krochko et al., 1998; Cutler and
Krochko, 1999). Similar to PA, neophaseicacid (neoPA) can be formed from
hydroxy ABA through isomerization (Zhou et al., 2004). ABA 8-hydroxy-
lases are members of a CYP707A subfamily of cytochrome P450 monooxy-
genases (Kushiro et al., 2004; Saito et al., 2004). It has been reported that
exposure of plants to low VPD induces catabolism of ABA via CYP707As
genes (Okamoto et al., 2009).
Apart from the oxidative catabolic pathways, ABA can be inacti-
vated via conjugation with glucose to form its glucose ester (ABA-GE)

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(Xu et al., 2002; Priest et al., 2006). ABA-GE is readily reversible but
not easily permeable to biomembranes and may function as a realizable
and transportable form of ABA (Dietz et al., 2000; Sauter et al., 2002).
When ABA is needed, ABA-GE is hydroxylated through β-glucosidase
to increase the active form of ABA (Lee et al., 2006). The activity of
β-glucosidase was decreased in rose plants that had been grown in low
VPD compared with its activity in moderate-VPD-grown plants. This
resulted in a higher ratio of ABA-GE to ABA (Arve et al., 2012). However,
it is still not clear whether the lower ABA level after long-term exposure
to low VPD is due to lower production or the higher catabolism of ABA.
It has been shown that exposure to different VPDs influences ABA in
the leaves of plants. In many plant species, such as spinach (Spinacia olera-
cea) (Zeevaart, 1974), rose (Arve et al., 2012; Giday et al., 2013a), Arabidopsis
(Okamoto et al., 2009), and spiderwort (Tradescantia virginiana) (Rezaei
Nejad and van Meeteren, 2007, 2008), the foliar [ABA] decreases as a result
of exposure to low-VPD conditions. In Tradescantia, 1 day after transfer-
ring moderate-VPD-grown plants to a low-VPD condition, the foliar [ABA]
decreased to the ABA level found in plants grown at low VPD. Reciprocal
transfer of moderate-VPD-grown plants back from low to moderate VPD
increased the foliar [ABA] again to its original level in moderate-VPD-
grown plants (Rezaei Nejad and van Meeteren, 2008). In rose plants, con-
trary to moderate-VPD-grown plants, exposure of low-VPD-grown plants
to darkness did not result in elevation of the foliar [ABA] (Arve et al., 2012;
Giday et al., 2013a). In Arabidopsis, the [ABA] decreased sharply even 1 h
after exposure to a low-VPD condition (Okamoto et al., 2009). Moreover,
in vitro–propagated plants that were produced under low-VPD conditions
were deficient in ABA (Hronková et al., 2003). Therefore, in the absence of
stresses, the foliar [ABA] depends on the VPD as well; decreasing the VPD
will result in a decrease in [ABA].
A foliar threshold level for ABA has been suggested in order to keep
the stomata responsive to closing stimuli. Since the foliar ABA level in mod-
erate-VPD-exposed plants is always higher than this threshold, their sto-
mata are always responsive. In different rose cultivars and also Arabidopsis
accessions, higher ABA levels than the threshold were still present after
low-VPD exposure; these genotypes maintained responsiveness to closing
stimuli in the postharvest phase (Giday et al., 2013a; Aliniaeifard and van
Meeteren, 2014). In genotypes that showed malfunctioning of stomata after
a 4-day exposure to low VPD, a daily spray of ABA during the 4-day expo-
sure to low VPD resulted in maintaining the stomatal closing response to
closing stimuli in Arabidopsis and bean (Aliniaeifard and van Meeteren,
2013, 2014; Aliniaeifard, 2014; Aliniaeifard et al., 2014). This confirmed that
a long-term low ABA level results in loss of stomatal closing ability after-
wards. Changes in the signaling pathway inside the guard cells of stomata
due to long-term exposure to low-VPD conditions are reviewed in detail by

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Aliniaeifard and van Meeteren (2013). They proposed that alterations in

signaling pathways due to a long-term low transpiration rate under long-
term exposure to environmental conditions, especially low VPD, lead to
depletion of ABA, Ca 2+, and H 2O2 in the guard cells, as well as depletion of
extracellular ABA and Ca 2+. This will be accompanied by a low sensitivity
for ABA, due to a strong negative regulation of the ABA signaling path-
way by the type 2C protein phosphatases ABI1 and ABI2 and a low positive
regulation of the ABA signaling pathway by OST1. These effects will be
strengthened by a low sensitivity of the anion channel SLAC1 for Ca 2+. This
coincidence in changes of Ca 2+, ABA receptors, and positive and negative
regulators of ABA signaling is proposed as an explanation for persistency
of the stomatal malfunctioning induced by long-term exposure to low VPD.

6.4.1.2  Induction of Stomata Morphological

Changes by Low VPD
Besides stomatal closing ability, stomatal size and density can also be
influenced by VPD (Fordham et al., 2001; Tricker et al., 2012). Rose,
Tradescantia, and bean plants that developed their leaves in low-VPD
conditions are characterized by large stomata and a wide aperture area
(Figure  6.3) (Torre et al., 2003; Rezaei Nejad and van Meeteren, 2005;
Fanourakis et al., 2011, 2013a; Aliniaeifard et al., 2014). A question arises:
Are stomatal morphological alterations involved in the decreased ­stomatal
closing ­
ability of low-VPD-grown plants after harvest? A connection
between stomatal function and structural features for various species
has previously been suggested (Franks and Farquhar, 2007; Doheny-
Adams et al., 2012; Drake et al., 2013; Giday et al., 2013b). Because of the
higher ratio of a guard cell’s membrane surface to its volume, species with
smaller stomata may respond faster than species with larger stomata. On

L M

100 μm 100 μm

Figure 6.3  Stomatal size and density in low (L)- and moderate (M)-VPD-
grown Vicia faba plants.

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the other hand, smaller stomata are usually associated with higher stoma-
tal density per leaf area (Hetherington and Woodward, 2003). To optimize
the trade-off between carbon gain and transpirational water loss, these
characteristics (smaller stomata and higher stomatal density) allow the
leaf to attain high stomatal conductance under favorable conditions, and
to promptly reduce stomatal conductance when conditions are unfavor-
able, which help the plant cope with stress conditions (Xu and Zhou, 2008;
Doheny-Adams et al., 2012). Using five closely related species of the genus
Banksia, it has been demonstrated that the rate of stomatal response was
negatively correlated with stomatal size (Drake et al., 2013). It has been
indicated that smaller stomata require less leaf drying to close and that
plants’ stomatal size underlays much of the variation in the regulation of
transpiration upon desiccation (postharvest phase) after growth of the
plants in low and moderate VPDs (Franks and Farquhar, 2007; Doheny-
Adams et al., 2012; Drake et al., 2013; Giday et al., 2013b).
In cut rose flowers, although changes in stomatal length had no
influence on stomatal functionality, anatomical features per se represent a
significant and direct contribution to the increased water loss in the post-
harvest phase of the roses (Fanourakis et al., 2013a). Higher stomatal den-
sities enlarge the stomatal pore area per leaf area, leading to an increase
in the transpiration rate in low-VPD-grown plants. However, in faba bean,
stomatal density decreased as a result of growing them at low VPD. On
the other hand, in faba bean, stomata of low-VPD-grown plants were
larger for all guard cell dimensions than those of moderate-VPD-grown
plants (Aliniaeifard et al., 2014). Therefore, high stomatal conductance
in low-VPD-grown plants can be attributed to their larger stomata with
wider pore area in comparison with the stomata in m ­ oderate-VPD-grown
plants. In the previous studies, investigating the impact of stomatal mor-
phological traits on stomatal functionality, the stomatal responsiveness
of different species was investigated or plants from one or several geno-
types were exposed to contrasting environments, which usually induced
changes in both stomatal morphology and responsiveness (Hetherington
and Woodward, 2003; Torre et al., 2003; Franks and Farquhar, 2007; Drake
et al., 2013; Fanourakis et al., 2013a; Giday et al., 2013b). Analyzing the
importance of the stomatal morphological traits on the stomatal function-
ality, not only in moderate and low-VPD-grown plants, but also in plants
that had developed their leaves in moderate VPD and thereafter trans-
ferred for 1–4 days to low VPD, confirmed that stomata ­morphological
traits are not always determinant of stomatal functionality: the morphol-
ogy of stomata in plants that had developed their leaves at moderate VPD
and were then transferred for 4 days to low VPD was more similar to that
of ­moderate-VPD-grown plants, while their response to ABA and des-
iccation was similar to the stomatal response of low-VPD-grown plants
(Aliniaeifard et al., 2014).

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It has been suggested that changes in the signaling related to ABA

are the determining factor in the decreased closing ability of stomata and,
as a result, decreased vase or shelf life of horticultural crops after long-term
exposure to low VPD (Aliniaeifard and van Meeteren, 2013; Aliniaeifard,
2014; Aliniaeifard et al., 2014). It can be concluded that production of flow-
ers and vegetables in low VPD renders the stomata incapable of a suitable
response to closing stimuli afterwards. In the flowers, their rate of water
uptake becomes lower than their transpiration rate, resulting in a shorter
vase life due to a negative water balance, and leafy vegetables have a
reduced capacity to keep water after harvest, resulting in a loss of textural
and nutritional quality.

6.4.2 Temperature
As discussed in Section 6.2.1, stomata are involved in controlling plant
t emperature through the transpiration rate. Therefore, it could be
­
expected that stomata will open more with increasing temperatures to
prevent overheating of the plant. However, the response of stomata to tem-
perature is not constant for different temperatures, and usually it is species
dependent. There are contradictory reports regarding stomatal response
to different temperatures. Both stomatal closure and stomatal opening
have been reported as a result of an increase in temperature (Schulze
et al., 1973; Wilkinson et al., 2001). Likely this is because stomata opening
responds to many factors; among these factors are the CO2 concentration
in the stomatal cavity and the water potential of the leaves. These param-
eters are respectively affected by the rate of photosynthesis and by water
uptake and water loss. These processes are also largely influenced by
temperature. Schulze et al. (1973) showed for five species (Zygophyllum
dumosum, Artemisia herba-alba, Hamada scoparia, Reaumurian egevensis,
Prunus armeniaca) that stomata opened more in response to a gradual
increase in temperature (from 31.6°C to 39.8°C) at low water stress. This
response was independent of VPD. At high plant water stress, however,
the stomatal response was reversed; that is, the stomata closed when the
temperature was gradually increased. In leaf disks of legumes floating on
water, stomata opened in darkness by increasing the temperature from
20°C to 45°C (Feller, 2006; Reynolds-Henne et al., 2010), even when ABA
was applied. By using epidermal strips floating on water, the direct effect
of temperature on guard cells was studied, excluding the effects of pho-
tosynthesis and water loss (Rogers et al., 1979). They showed that stoma-
tal aperture increased by an increase of temperature from 10°C to 40°C,
while at 45°C the aperture was less than those obtained at 35°C and 40°C.
In the experiments mentioned, there was a fast recovery of stomata to its
original apertures when temperature was decreased again. This indicates

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that large aftereffects of cultivation temperature on postharvest behavior

are not expected.
There are some reports that stomatal density decreases with increas-
ing temperature (Beerling and Chaloner, 1993). Stomatal density of
Dodonaea, produced as cut foliage branches, showed a strong correlation
with seasonal average monthly day temperature. The cut branches showed
also strong seasonal variations in longevity, wilting after 1 week in winter,
while displaying a vase life of 3 weeks in summer. On day 1 of vase life, the
branch water status was positively correlated with vessel, stomata, and tri-
chome densities, and negatively correlated with vessel member length and
leaf thickness. However, on day 16, the branch water status was negatively
correlated only with vessel member length and diameter, implying a differ-
ent relative importance of anatomical parameters for surviving water stress
in the vase. The limited part that stomata number plays in the vase life of
cut flowers was discussed in Section 6.2.
Mostly, horticultural products are cooled down after harvest. Rapid
stomatal closure can be induced by exposure to low temperatures. However,
there is variation in the stomatal closure response between species after
exposure to low temperatures. For example, in response to cold (7°C), sto-
mata of Commelina communis are closed within 1 hour of transferring the
plants from 27°C to 7°C, whereas the stomata of tobacco (Nicotiana rustica)
are not capable of closure, which leads to uncontrolled water loss and wilt-
ing of the leaves after 5 hours of the transfer to low temperature (Wilkinson
et al., 2001). The rapid low-temperature-induced stomatal closure is likely
to be the result of a low-temperature increase in guard cell plasma mem-
brane calcium channel activity. Chilling-sensitive plants, such as tobacco,
do not show this increase in calcium uptake at low temperature (Wilkinson
et al., 2001). It is also possible that in chilling-sensitive produce, the mem-
branes of epidermal cells lose their semipermeability characteristics, and
thus their turgor, before guard cells lose them. In that case, the stomata
will also open. It can be concluded that the effect of temperature during
the postharvest phase on the function of stomata, as well as on keeping the
quality of plant produce, is highly species dependent. Both low and high
temperatures can influence the proper function of stomata and, as a result,
influence the quality of the products. The effect of preharvest temperature
on the role of stomata in keeping quality is very limited.

6.4.3 Light
Greenhouse crop production frequently makes use of supplementary light-
ing to improve plant productivity in periods of the year when natural irradi-
ance is low; on some occasions, continuous lighting is used (Mortensen
and Fjeld, 1998; Dodd et al., 2005; Pettersen et al., 2006; Velez-Ramirez

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et al., 2011). For example, in pot roses, extending the lighting period from
18 to 24 h per day shortened the time to flowering by 12% and the num-
ber of flowers increased by 34% (Pettersen et al., 2007). Although grow-
ing plants under continuous light has several advantages (e.g., reduction
of powdery mildew), it adversely influences the postharvest life of the cut
flowers (Mortensen and Fjeld, 1998; Mortensen and Gislerød, 1999, 2011)
due to poor stomatal closure under conditions of decreasing leaf water
potential and turgor (Slootweg and van Meeteren, 1991; Arve et al., 2012).
The vase life of the roses decreased by using supplemental lighting. The
vase life depends on the duration of exposure to light. The longer the light-
ing period during growth, the shorter the vase life afterwards (Mortensen
and Fjeld, 1998; Pettersen et al., 2006; Mortensen and Gislerød, 1999, 2011;
Arve et al., 2012). Extending the lighting period from 16 to 20 h decreased
the postharvest life in lemon balm, while it is not affected in basil; however,
24 h exposure to light resulted in decreased postharvest life in both herbs
(Islam et al., 2010). It has been shown that by decreasing the RH in the
growing units when plants are grown under continuous light, it is possible
to reduce the disability of stomata to close and, as a result, increase the vase
life of cut flowers (Mortensen and Fjeld, 1998; Mortensen and Gislerød,
1999, 2011; Pettersen et al., 2006; Arve et al., 2012). However, even in mod-
erate RH conditions, exposure to continuous light results in decreased vase
life of cut roses (Mortensen and Fjeld, 1998).
Exposure to light not only influences the sugar content through pho-
tosynthesis, but also can induce sugar accumulation because of osmotic
adjustment in the cells. For example, higher sugar levels have been found
in the rinds of mandarin fruits positioned outside of the tree canopy fac-
ing toward sunlight. Sugar accumulated (osmotic adjustment) in the rinds
of sunlight-exposed fruits because of dehydration of the rind’s cells due
to high transpiration via stomata of sun-exposed fruit surfaces (Yakushiji
et  al., 1998; Magwaza et al., 2013). Accumulation of sugar and other bio-
chemicals in the rinds of sun-exposed fruit during the growing season
improves their postharvest performance and decreases susceptibility to
rind breakdown (Magwaza et al., 2013).
The amount of healthy compounds of a plant can be influenced by
the lighting period and light intensity. Although light has no direct role
in the production of ascorbic acid (vitamin C), the duration and intensity
of light can influence the production of ascorbic acid in the leaf (Lee and
Kader, 2000). In general, the amount of ascorbic acid increases by light
(intensity and duration) and decreases in the dark period (Lee and Kader,
2000). On the other hand, plants that have been produced in a prolonged
lighting period could lose their vitamin C more easily after harvest because
of higher water loss via stomata after harvest; therefore, there should be
a balance for the lighting period for producing vitamin C before harvest
and preventing loss of vitamin C after harvest of leafy vegetables. However,

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research in which the effect of light during cultivation on vitamin C content

during the pre- and postharvest phases is lacking. Also, the amount of other
secondary metabolites can be affected by light. For example, linalyl acetate
is one of the compounds in the essential oil of oregano plants with anti-
inflammatory activity (Peana et al., 2002). It has been found that growing
oregano plants in full light resulted in a decreased amount of linalyl acetate
due to more open stomata and higher transpiration rate. On the other hand,
growing plants in pots under 50% shade resulted in a higher percentage of
linalyl acetate in the essential oil (Tibaldi et al., 2011).

6.5  Stomata and Tolerance to Postharvest

Diseases and Physiological Disorders
Postharvest diseases reduce the quantity and quality of horticultural prod-
ucts. Although some diseases may not lower produce saleability, they still
reduce product value. Losses in value of the product through postharvest
diseases may occur at any time from harvest to consumption (Coates and
Johnson, 1997). The entrance of pathogens to plant tissues is the first and
critical stage during the infection process. Some pathogenic and sapro-
phytic microorganisms are able to survive and reduplicate on the leaf sur-
face without infection (Melotto et al., 2008). Although many of pathogenic
fungi can penetrate the cuticle and epidermis by different means (Mendgen
et al., 1996), some fungal diseases and many pathogenic bacteria are unable
to directly penetrate the host cuticle. The cuticle plays a key role in the
survival of plants against biotic and abiotic stresses. The primary physi-
ological role of the plant cuticle is to seal the tissue against a relatively dry
atmosphere, preventing desiccation by minimizing water loss (Mintz-Oron
et al., 2008; Riederer and Muller, 2008); the position of stomata in the inter-
face between the surrounding environment and leaf internal tissues adds
another role for this microscopic pore. It provides a route for endophytic
colonization of pathogens in the plant tissues; therefore, plants also have
established a mechanism to regulate stomatal aperture not only in response
to a range of environmental factors, but also in response to pathogens
(Melotto et al., 2008; Gudesblat et al., 2009). Both stomatal density and
size are important factors determining the infection of products by patho-
gens. Asiatic citrus canker caused by Xanthomonas citri subsp. citri seri-
ously affects the citrus industry in Asia. It causes formation of pustule-like
necrotic lesions on the fruit, and it highly decreases fruit quality. Stomatal
characteristics such as stomatal density, size, and closing response will
influence the magnitude of infection of fruit internal tissues. Stomatal char-
acteristics can be considered the main factor influencing the resistance
magnitude of citrus plants to the bacteria (Wang et al., 2011a). Therefore,

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the presence of more stomata and a larger stomatal aperture area in the fruit
surface could increase the susceptibility to bacteria. Wang et al. (2011a)
found that under the same experimental conditions, ‘Meiwa’ kumquat has a
lower stomatal density and smaller stomatal openings than ‘Newhall’ navel
orange. These stomatal characteristics of ‘Meiwa’ make this cultivar better
resistant to canker than ‘Newhall’. ‘Pinalate’ (Citrus sinensis L. Osbeck) is a
yellow ABA-deficient mutant derived from the orange ‘Navelate’; therefore,
its stomata will close less. ‘Pinalate’ has a higher transpiration rate, and due
to the impaired stomatal functioning, it is more susceptible to pathogenic
infections (Alférez et al., 2005).
Since stomata are the entrance site of many bacterial and fungal
diseases into leaves or fruits of horticultural products, the immunity of a
product to pathogen virulence in many cases depends on the function of
the stomata (Zeng et al., 2010). Many postharvest treatments are efficient
when they restrict the entrance of pathogens into the products. For exam-
ple, heat treatments such as hot water dips, vapor heat, short hot water
rinse, and brushing, causing melting of the wax layer in the surface of the
fruits, and therefore covering of the stomata, provide a mechanical barrier
for the pathogens, restricting the invasion of pathogens into the internal
tissues (Porat et al., 2000; Schirra et al., 2000; Waller et al., 2001; Mulas
and Schirra, 2007). On some occasions, exogenous wax is applied in order
to extend the quality of the fruits. The amount of wax applied on the fruit is
important to prevent pathogen infection. Low-coating loads result in some
uncoated surface areas with bare stomata, while sufficient wax loads cover
all the stomata and restrict pathogen infections (Njombolwana et al., 2013).
Salicylic acid is one of the plant hormones known to be involved in sensing
the pathogenic invasion. Salicylic acid–induced stomatal closure is one of
the cellular pathways that has been suggested after infection by pathogens
(Lu and Chen, 2005; Melotto et al., 2006; Khokon et al., 2011). It has been
shown that inducing stomatal closure by salicylic acid improves the plant
capacity to cope with diseases. For example, in lily, salicylic acid treatments
lead to induction of stomatal closure, causing more tolerance of lily plants to
Botrytis elliptica (Lu and Chen, 2005).
In some products, monitoring stomatal conductance can be used
as a nondestructive index for storability and quality status. Controlling
water loss from the leaves (especially from the stomata) can be consid-
ered an important way to minimize quality problems and extend the
postharvest life in crops such as rose cut flowers and kalanchoe cuttings
(Bredmose and Nielsen, 2009; Fanourakis et al., 2013a, 2013b). Stomatal
density in tomato fruits is very low, whereas the number of stomata in
cucumber is relatively high (Table 6.2). Higher stomatal densities posi-
tively correlate with higher water loss from the fruits. Measuring the
amount of water loss immediately after harvest showed 20 times higher

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water loss for cucumber fruits than tomato fruits (Adams and Ho, 1995).
Therefore, fast handling after harvest is important in order to restrict
water loss and quality loss in crops with higher stomatal densities. High
humidity during growth decreases movement of calcium to the leaves in
cucumber and tomato plants. However, due to the higher stomatal den-
sity in the fruits of cucumber than in tomato fruits, a higher level of cal-
cium accumulates in the cucumber fruits (Adams and Ho, 1995). This
confirmed that transpiration by fruits plays a pivotal role in determining
calcium levels in the fruits and susceptibility to physiological disorders
related to calcium deficiency.

6.6  Postharvest Treatments and Stomata

In many horticultural products, the stomata are functional after harvest,
and their function is important for postharvest quality (Johnson and Brun,
1966; Moreshet and Green, 1980; Blanke and Lenz, 1989; Burg, 2004).
Storage of fresh-cut Romaine lettuce in light conditions increases the num-
ber of open stomata and results in increased water loss and increased occur-
rences of leaf browning (Martínez-Sánchez et al., 2011). Stomatal opening
in Chinese kale positively correlates with loss of fresh weight. However,
vitamin C content in light-stored Chinese kale leaves was higher than in
dark-stored leaves (Noichinda et al., 2007).
Postharvest potassium silicate application covers fruit stomata with a
silicate layer, which positively delays fruit weight loss by keeping fruit mois-
ture (Tesfay et al., 2011). In some horticultural products, postharvest heat
treatments decrease water loss from the product instead of increasing it.
This effect of postharvest heat treatments is due to their effects on the melt-
ing of cuticular wax, which causes coverage of stomata and microcracks;
therefore, heat treatments are able to reduce water loss by decreasing tran-
spiration (Schirra et al., 2000). For example, treatment of Valencia oranges
by 45°C water for 42 min keeps the firmness of the fruit and considerably
decreases water loss. But immersion of fruits in 53°C water for 12 minutes
increases the water loss from the fruits (Williams et al., 1994; Tang et al.,
2007). However, by shortening the time of hot water immersion, it would
be possible to use hotter water to control water loss. For example, treat-
ing orange fruits at 56°C for 20 s also melts epicuticular waxes, leading to
covered and sealed stomata and cracks on the fruit without occurrences of
other related problems, such as fruit weight loss, surface damage, or inter-
nal quality loss (Porat et al., 2000).
Postharvest pathogens are one of the items considered for loss of
quality during storage of the products. Since the main ports for entrance
of pathogenic disease are stomata, covering of stomata by wax has the
potential to reduce the incidence of postharvest diseases by providing a

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physical barrier against pathogens (Schirra et al., 2000; Li et al., 2013).

Blocking stomata and other cracks by treating orange fruits at 56°C for
20 s, also restricts occurrences of Penicillium digitatum (Porat et al., 2000).
Although ethylene can induce some quality problems in the postharvest
stage (hastening ripening or senescence), treatment with ethylene forms
new waxes on the stored fruits that cover stomata, cracks, or areas lacking
wax and provides mechanical barriers to P. digitatum penetration (Cajuste
et al., 2010). As another role in postharvest treatments, the presence of
stomata facilitates the absorption of 1-methylcyclopropene (1-MCP) into
product tissues (Dong et al., 2013), a gas often used to inhibit senescence
as it blocks ethylene receptors.
Storage of products in low-pressure-type storage induces stomatal
malfunctioning in response to darkness (Laurin et al., 2006). The problem
with stomatal malfunctioning worsens when the product is stored at low
pressure for a long time period. Under such conditions, it has been reported
that the marketable period shortens and the weight loss of cucumber
increases by 5% (Laurin et al., 2006). Similarly, in grape berries, 20% of the
stomata stay fully or partially open at darkness (Blanke and Leyhe, 1988).
Kirk et al. (1986) reported that long-term storage of fresh hibiscus cuttings
in low-pressure storage induces stomatal malfunctioning in response to
several closing stimuli, such as high carbon dioxide and low RH.
The stomatal closing effect of ABA has been used to control water loss in
order to extend the postharvest life of cut flowers and potted plants (Cornish
et al., 1985; Pompodakis et al., 2004; Shimizu-Yumoto and Ichimura, 2009;
Reid and Jiang, 2012). Since ABA is an expensive compound, some stud-
ies have tried to find a way to increase the endogenous ABA level with less
expensive methods. In some studies, NaCl or osmotic stress was applied to
the plants to close the stomata in order to extend their postharvest life (Reid
and Jiang, 2012; Giday et al., 2014). S-Abscisic acid (S-ABA), the biologically
active form of ABA produced through microbial fermentation, is commer-
cially used for extending postharvest life of products. However, in some spe-
cies, some damage to the flower and shoot has been reported by using this
compound (Runkle et al., 2007; Waterland et al., 2010). It seems that using a
high concentration of S-ABA is problematic, while using low concentrations
can extend postharvest life without negative effects (Reid and Jiang, 2012).
However, senescence can be enhanced by application of ABA (Halevy et al.,
1974; Ferrante et al., 2004; Hunter et al., 2004).

6.7  Ethylene, Stomata, and Senescence

To integrate all the plant responses to the environment, phytohormones play
an important role, as well as the interplay between them. The role of ABA
as the main phytohormone reducing transpiration by provoking stomatal

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

closure is mentioned above. There are contradictory reports regarding sto-

matal responses to ethylene. Both induction of stomatal closure (Vitagliano
and Hoad, 1978; Pallas and Kays, 1982; Taylor and Gunderson, 1986, 1988;
Tissera and Ayres, 1986; Desikan et al., 2005, 2006) and promotion of sto-
matal opening (Frommhold, 1982; Merritt et al., 2001; She and Song, 2012;
Song et al., 2012) have been reported for ethylene. However, in some stud-
ies, no effect of ethylene on stomatal opening and closure has been reported
(Pallaghy and Raschke, 1972; Bradford, 1983; Madhavan et al., 1983). The
interplay between ethylene and other phytohormones is likely to be determi-
nant in the regulation of stomatal movements by ethylene (Pallas and Kays,
1982; Merritt et al., 2001; Tanaka et al., 2005, 2006; Acharya and Assmann,
2009; Chen et al., 2013). It has been reported that induction of stomatal open-
ing or inhibition of stomatal closure by auxins and cytokinins is through their
promotional effect on ethylene production (Merritt et al., 2001; Tanaka et al.,
2006). It seems that stomata close in response to ethylene in the absence of
ABA and open in the presence of ABA (Desikan et al., 2006; Tanaka et al.,
2006). It has been shown that ethylene or ACC (the precursor of ethylene)
can prevent ABA-induced stomatal closure (Tanaka et al., 2005). Application
of ethylene increases stomatal aperture of wild-type Arabidopsis exposed to
drought conditions (Tanaka et al., 2005). It is likely that ethylene acts on the
late steps of the ABA-induced stomatal closure (Tanaka et al., 2005). The
interaction between AtERF7 (which is a member of the ethylene-responsive
binding factor) and PKS3 (a Ser/Thr protein kinase that interacts with ABI2
and, to some extent, ABI1) (Guo et al., 2002) decreases the guard cells’ sen-
sitivity to ABA and increases water loss through transpiration (Song et al.,
2005). For induction of stomatal closure by ethylene, it seems that ethyl-
ene close the stomata through an ABA-independent pathway. The ethylene
receptor ETR1 mediates H 2O2 signaling in guard cells and may be a sensor
for H 2O2 perception in guard cells (Desikan et al., 2005). So ETR1 func-
tions as the site of ethylene and H 2O2 cross talk, leading to stomatal closure
(Desikan et al., 2005).
Usually after harvesting, horticultural products are stored in dark-
ness and face water stress and high CO2 concentrations during storage. It
has been shown that water stress and high CO2 concentrations can induce
ethylene production. Stomatal responses are sensitive to darkness, water
stress, and CO2. As discussed before, it seems that the effect of ethylene
on stomatal closure highly depends on other factors, such as the presence
of other stresses and its interaction with other phytohormones. In order to
keep the quality of a product during storage, it is important to know the spe-
cies-specific stomatal responses of the product to CO2 and ethylene. Usually
the stomata of harvested leaves close instantaneously or within 10–35 min
because of harvesting procedures, and this considerably increases their
internal ethylene concentrations (Burg, 2004). On the other hand, CO2 can
inhibit the enhancing effect of ethylene on senescence (Aharoni et al., 1979;

188
 Stom ata a n d P ostharvest P hy s i o l o g y

Sisler and Wood, 1988). The dual effect of high CO2 during storage is pre-
sented in Figure 6.4. At atmospheric pressure, closure of stomata occurs
during storage and transport in darkness, but using low-pressure (LP)
storage encourages stomatal opening even in darkness (Kirk and Skytt
Anderson, 1985; Kirk et al., 1986; Veierskov and Kirk, 1986). This low sto-
matal resistance at LP results in improved gas diffusion (such as ethylene)
and considerably improves intercellular ventilation. In Valencia orange with
open stomata, the transpirational resistance is 13.5 scm−1 (Moreshet and
Green, 1980). In the fruits, the stomatal resistance to ethylene transport is
23 scm−1. Due to stomatal closure during harvest, the resistance to ethyl-
ene transport increases to 6886 scm−1 in the orange fruits (Ben Yehoshua
et al., 1979). Moreover, fruit’s cuticular resistance to ethylene transport is
approximately 10-fold higher than its diffusive resistance to ethylene of
the stomata. Under the situation of closed stomata, the internal ethylene
concentration largely increases. Therefore, in order to prevent detrimental
effects of ethylene, it is vital to decrease the internal ethylene concentration
by some means, such as vacuum extraction (Burg, 2004).
Detaching organs from the mother plants and placing them in dark-
ness can promote leaf senescence in many plant species. A relationship
between stomatal aperture and senescence has been suggested (Thimann

High CO2 Darkness Desiccation

Stomatal closure

After
High internal harvest
ethylene level

Accelerated
senescence
(quality loss)

Figure 6.4  Diagram describing the effect of stomatal closing stimuli

­prevalent after harvest on the product quality.

189
P OST H AR V EST RI P ENING P H Y SIOLOG Y

and Satler, 1979). Senescence can be accelerated in darkness and slowed

down in light. The preventive effect of light on senescence is partly due to
its effect on stomatal opening, as suggested by the finding that promotion of
stomatal opening in darkness (using cytokinins, fusicoccin, etc.) postpones
senescence (in a similar way as light) (Thimann and Satler, 1979; Thimann,
1985). Cytokinins induce stomatal opening and delay senescence (Gan and
Amasino, 1995; Wingler et al., 1998; Song et al., 2006; Tanaka et al., 2006).
On the other hand, it has been shown that senescence can be accelerated by
ethylene. As discussed before, there is a controversy in reports regarding the
role of ethylene on stomatal closure. The rate of senescence level correlates
positively with the ABA level. Since antagonistic effects of cytokinins on ABA
have been proved, this can explain the preventive effects of cytokinins on
senescence (Thimann, 1985; Pospíšilová et al., 2000; Wang et al., 2011b).
In the flowers (petals) also, ABA regulates the process of senescence.
The level of ABA increases as senescence of petals starts. Also, a higher
ABA level has been shown in short-lived rose genotypes than in long-lived
genotypes (Mayak et al., 1972; Swanson et al., 1975). Confirming the role
of ABA in senescence, it has been reported that exogenous application of
ABA accelerates senescence in carnations (Mayak and Dilley, 1976) and
roses (Mayak et al., 1972).
The effect of the environment on leaf senescence is local, and it dif-
fers from young leaves to old leaves (leaf age dependency) (Weaver and
Amasino, 2001). It has been shown that the problem of low-VPD-induced
stomatal malfunctioning increases by leaf age; after exposure to low VPD,
stomata of old leaves are less responsive to desiccation than the stomata
of younger leaves (Rezaei Nejad and van Meeteren, 2007). In Arabidopsis,
after desiccation the rate of water loss increases by the leaf age; the expres-
sions of Senescence-Associated Gene113 (SAG113) and AtNAP (gene encod-
ing for a NAC family transcription factor, AtNAP) are co-induced during
leaf senescence (Zhang and Gan, 2012; Zhang et al., 2012). SAG113, a gene
that encodes a protein phosphatase belonging to the PP2C family, functions
as a negative regulator of the ABA signaling pathway and prevents stomatal
closure in response to closing stimuli such as ABA and desiccation (Zhang
et al., 2012). The AtNAP transcription factor physically interacts with the
promoter region of SAG113 and promotes its expression at the transcrip-
tional level (Zhang and Gan, 2012) and keeps the stomata less functional to
closing stimuli.

6.8  Conclusions and Future Perspectives

Climatic conditions in both protected and open-field cultivation can influ-
ence postharvest performance of horticultural products. During posthar-
vest handling and storage, many parameters, such as species, variety, light,

190
 Stom ata a n d P ostharvest P hy s i o l o g y

humidity, temperature, radiation exposure, microbial load, presence of

pests, packaging, and chemical treatments, considerably change physical
and biochemical properties and the rate of deterioration of horticultural
products. Sometimes, growing plants in certain conditions results in the
optimum growth of the plants, but after harvest, several problems for the
product occur that lead to decreased shelf or vase life and, in the worse situ-
ations, deterioration of horticultural products.
Stomata are the main ports connecting internal leaf space to the out-
side environment. They can play an important role in connecting prehar-
vest factors to postharvest quality changes; this possible linkage function
has hardly been investigated.

References
Aarts, J.F.T. 1957. Over de houdbaarheid van snijbloemen [On the keepabil-
ity of cut flowers]). Mediterrenean Landbouwhogeschool Wageningen 57,
1–62.
Acharya, B.R., and Assmann, S.M. 2009. Hormone interactions in stomatal
function. Plant Molecular Biology 69, 451–462.
Adams, P., and Ho, L. 1995. Differential effects of salinity and humidity on
growth and Ca status of tomato and cucumber grown in hydroponic
culture. Acta Horticulturae 401, 357–364
Agüero, M.V., Ponce, A.G., Moreira, M.R., and Roura, S.I. 2011. Lettuce qual-
ity loss under conditions that favor the wilting phenomenon. Postharvest
Biology and Technology 59, 124–131.
Aguilar, M.L., Espadas, F.L., Coello, J., Maust, B.E., Trejo, C., Robert, M.L.,
and Santamaría, J.M. 2000. The role of abscisic acid in controlling leaf
water loss, survival and growth of micropropagated Tagetes erecta
plants when transferred directly to the field. Journal of Experimental
Botany 51, 1861–1866.
Aharoni, N., Anderson, J.D., and Lieberman, M. 1979. Production and action
of ethylene in senescing leaf discs effect of indoleacetic acid, kinetin,
silver ion, and carbon dioxide. Plant Physiology 64, 805–809.
Alférez, F., Sala, J.M., Sanchez-Ballesta, M.T., Mulas, M., Lafuente, M.T., and
Zacarias, L. 2005. A comparative study of the postharvest performance
of an ABA-deficient mutant of oranges. I. Physiological and quality
aspects. Postharvest Biology and Technology 37, 222–231.
Aliniaeifard, S. 2014. Signal transduction pathways in guard cells after
prolonged exposure to low vapour pressure deficits. PhD thesis,
Wageningen University.
Aliniaeifard, S., and van Meeteren, U. 2013. Can prolonged exposure to
low VPD disturb the ABA signaling in stomatal guard cells? Journal of
Experimental Botany 64, 3551–3566.

191
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Aliniaeifard, S., and van Meeteren, U. 2014. Natural variation in stomatal

response to closing stimuli among Arabidopsis thaliana accessions after
exposure to low VPD as a tool to recognise the mechanism of disturbed
stomatal functioning. Journal of Experimental Botany, doi: 10.1093/jxb/
eru370.
Aliniaeifard, S., Malcolm Matamoros, P., and van Meeteren, U. 2014. Stomatal
malfunctioning under low VPD conditions: Induced by morphological
and anatomical or by signaling alterations? Physiologia Plantarum 152,
688–699.
Amjad, M., Akhtar, J., Anwar-ul-Haq, M., Yang, A., Akhtar, S.S., and Jacobsen,
S.E. 2014. Integrating role of ethylene and ABA in tomato plants adapta-
tion to salt stress. Scientia Horticulturae 172, 109–116.
Antoni, R., Rodriguez, L., Gonzalez-Guzman, M., Pizzio, G.A., and Rodriguez,
P.L. 2011. News on ABA transport, protein degradation, and ABFs/
WRKYs in ABA signaling. Current Opinion in Plant Biology 14, 547–553.
Aracama, C.V., Kane, M.E., Wilson, S.B., and Philman, N.L. 2008. Comparative
growth, morphology, and anatomy of easy- and difficult-to-acclimatize
sea oats (Uniola paniculata) genotypes during in vitro culture and ex
vitro acclimatization. Journal of the American Society for Horticultural
Science 133, 830–843.
Arve, L.E., Terfa, M.T., Gislerød, H.R., Olsen, J.E., and Torre, S. 2012. High
relative air humidity and continuous light reduce stomata functional-
ity by affecting the ABA regulation in rose leaves. Plant, Cell and
Environment 36, 382–392.
Atkins, C.A., Kuo, J., Pate, J.S., Flinn, A.M., and Steele, T.W. 1977.
Photosynthetic pod wall of pea (Pisum sativum L.) distribution of car-
bon dioxide-fixing enzymes in relation to pod structure. Plant Physiology
60, 779–786.
Azad, A.K., Sawa, Y., Ishikawa, T., and Shibata, H. 2007. Temperature-dependent
stomatal movement in tulip petals controls water transpiration during
flower opening and closing. Annals of Applied Biology 150, 81–87.
Bakker, J.C. 1991. Effects of humidity on stomatal density and its relation to
leaf conductance. Scientia Horticulturae 48, 205–212.
Barbieri, G., Vallone, S., Orsini, F., Paradiso, R., De Pascale, S., Negre-
Zakharov, F., and Maggio, A. 2012. Stomatal density and metabolic
determinants mediate salt stress adaptation and water use efficiency in
basil (Ocimum basilicum L.). Journal of Plant Physiology 169, 1737–1746.
Barth, M.M., Perry, A.K., Schmidt, S.J., and Klein, B.P. 1992. Misting affects
market quality and enzyme activity of broccoli during retail storage.
Journal of Food Science 57, 954–957.
Bauer, H., Ache, P., Lautner, S., Fromm, J., Hartung, W., Al-Rasheid,
Khaled  A.S., Sonnewald, S., Sonnewald, U., Kneitz, S., Lachmann, N.,
Mendel, R.R., Bittner, F., Hetherington, A.M., and Hedrich, R. 2012. The
stomatal response to reduced relative humidity requires guard cell-
autonomous ABA synthesis. Current Biology 23, 53–57.

192
 Stom ata a n d P ostharvest P hy s i o l o g y

Beerling, D.J., and Chaloner, W.G. 1993. The impact of atmospheric CO2 and
temperature-change on stomatal density—observations from quercus-
robur lammas leaves. Annals of Botany 71, 231–235.
Belin, C., De Franco, P.O., Bourbousse, C., Chaignepain, S., Schmitter, J.M.,
Vavasseur, A., Giraudat, J., Barbier-Brygoo, H., and Thomine, S. 2006.
Identification of features regulating OST1 kinase activity and OST1
function in guard cells. Plant Physiology 141, 1316–1327.
Ben-Yehoshua, S., and Rodov, V. 2003. Transpiration and water stress. In
Postharvest Physiology and Pathology of Vegetables, ed. J.A. Bartz and J.K.
Brecht, 111–159. 2nd ed. Marcel Dekker, New York.
Ben-Yehoshua, S., Kobiler, I., and Shapiro, B. 1979. Some physiological
effects of delaying deterioration of citrus fruits by individual seal pack-
aging in high density polyethylene film. Journal of the American Society
for Horticultural Science 104, 868–872.
Black, C.C., and Osmond, C.B. 2005. Crassulacean acid metabolism pho-
tosynthesis: ‘working the night shift’. In Discoveries in Photosynthesis,
ed. Govindjee, J.T. Beetty, H. Gest, and J.F. Allen, 881–893. Springer,
Dordrecht, The Netherlands.
Blanke, M. 1986. Comparative SEM study of stomata on developing quince,
apple, grape and tomato fruit. Angewandte Botanik 60, 209–214.
Blanke, M. 1987. Comparative SEM study of stomata and surface morphol-
ogy in apple. Angewandte Botanik 61, 433–438.
Blanke, M. 2002. Photosynthesis of strawberry fruit. Acta Horticulturae 567,
373–376.
Blanke, M., and Lenz, F. 1989. Fruit photosynthesis. Plant, Cell and
Environment 12, 31–46.
Blanke, M.M. 1992. Photosynthesis of avocado fruit. In Proceedings of Second
World Avocado Congress, vol. 1, pp. 179–189. University of California,
Davis, CA.
Blanke, M.M. 1994. Regulation of respiration in apple, avocado and citrus
orange fruit. Acta Horticulturae 398, 139–146.
Blanke, M.M., and Leyhe, A. 1988. Stomatal and cuticular transpiration of the
cap and berry of grape. Journal of Plant Physiology 132, 250–253.
Blanke, M.M., Höfer, M., and Pring, R.J. 1994. Stomata and structure of tetra-
ploid apple leaves cultured in vitro. Annals of Botany 73, 651–654.
Blatt, M.R. 2000. Cellular signaling and volume control in stomatal movements
in plants. Annual Review of Cell and Developmental Biology, 16, 221–241.
Bohnert, H.J., Nelson, D.E., and Jensen, R.G. 1995. Adaptations to environ-
mental stresses. Plant Cell 7, 1099–1111.
Bradford, K.J. 1983. Involvement of plant growth substances in the altera-
tion of leaf gas exchange of flooded tomato plants. Plant Physiology 73,
480–483.
Brainerd, K.E., and Fuchigami, L.H. 1982. Stomatal functioning of in vitro and
greenhouse apple leaves in darkness, mannitol, ABA, and CO2. Journal
of Experimental Botany 33, 388–392.

193
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Bredmose, N.B., and Nielsen, K.L. 2009. Controlled atmosphere storage

at high CO2 and low O2 levels affects stomatal conductance and influ-
ence root formation in kalanchoe cuttings. Scientia Horticulturae 122,
91–95.
Burchi, G., and Prisa, D. 2013. Preharvest conditions that can improve the
postharvest quality of ornamentals. Acta Horticulturae 970, 23–28.
Burg, S.P. 2004. Postharvest Physiology and Hypobaric Storage of Fresh Produce.
CABI Publishing, Wallingford, UK.
Cajuste, J.F., González-Candelas, L., Veyrat, A., García-Breijo, F.J., Reig-
Armiñana, J., and Lafuente, M.T. 2010. Epicuticular wax content
and morphology as related to ethylene and storage performance of
‘Navelate’ orange fruit. Postharvest Biology and Technology 55, 29–35.
Camejo, D., Rodríguez, P., Angeles Morales, M., Miguel Dell’Amico, J.,
Torrecillas, A., and Alarcón, J.J. 2005. High temperature effects on pho-
tosynthetic activity of two tomato cultivars with different heat suscepti-
bility. Journal of Plant Physiology 162, 281–289.
Camposeo, S., Palasciano, M., Vivaldi, G.A., and Godini, A. 2011. Effect of
increasing climatic water deficit on some leaf and stomatal parame-
ters of wild and cultivated almonds under Mediterranean conditions.
Scientia Horticulturae 127, 234–241.
Ceulemans, R., Heursel, J., Ibrahim, N., and Impens, I. 1984. Variations
among physiological, morphological and biochemical characteristics
of evergreen azalea (Rhododendron simsii Planch.) cultivars. Scientia
Horticulturae 22, 147–155.
Chen, L., Dodd, I.C., Davies, W.J., and Wilkinson, S. 2013. Ethylene limits
abscisic acid- or soil drying-induced stomatal closure in aged wheat
leaves. Plant, Cell and Environment 36, 1850–1859.
Chen, X., Pierik, R., Peeters, A.J.M., Poorter, H., Visser, E.J.W., Huber, H., de
Kroon, H., and Voesenek, L.A.C.J. 2010. Endogenous abscisic acid as a
key switch for natural variation in flooding-induced shoot elongation.
Plant Physiology 154, 969–977.
Cheng, W.H., Endo, A., Zhou, L., Penney, J., Chen, H.C., Arroyo, A., Leon, P.,
Nambara, E., Asami, T., and Seo, M. 2002. A unique short-chain dehy-
drogenase/reductase in Arabidopsis glucose signaling and abscisic acid
biosynthesis and functions. Plant Cell Online 14, 2723–2743.
Chimona, C., Stamellou, A., Argiropoulos, A., and Rhizopoulou, S. 2012. Study
of variegated and white flower petals of Capparis spinosa expanded at
dusk in arid landscapes. Journal of Arid Land 4, 171–179.
Christensen, J.V. 1976. Cracking in cherries. Danish Journal of Plant and Soil
Science 80, 289–324.
Christmann, A., Grill, E., and Huang, J. 2013. Hydraulic signals in long-dis-
tance signaling. Current Opinion in Plant Biology 16, 293–300.

194
 Stom ata a n d P ostharvest P hy s i o l o g y

Christmann, A., Hoffmann, T., Teplova, I., Grill, E., and Muller, A. 2005.
Generation of active pools of abscisic acid revealed by in vivo imaging
of water-stressed arabidopsis. Plant Physiology 137, 209–219.
Christmann, A., Weiler, E.W., Steudle, E., and Grill, E. 2007. A hydraulic
signal in root-to-shoot signaling of water shortage. Plant Journal 52,
167–174.
Coates, L., and Johnson, G. 1997. Postharvest diseases of fruit and vegeta-
bles. In Plant Pathogens and Plant Diseases, ed. J.F. Brown, and H.J.
Ogle, 533–548. Rockvale Publications, Armidale, Australia.
Cornish, K., King, A., Reid, M., and Paul, J. 1985. Role of ABA in stress-
induced reduction of water loss from potted chrysanthemum plants.
Acta Horticulturae 167, 381–386.
Crawford, A.J., McLachlan, D.H., Hetherington, A.M., and Franklin, K.A.
2012. High temperature exposure increases plant cooling capacity.
Current Biology 22, R396–R397.
Cutler, A.J., and Krochko, J.E. 1999. Formation and breakdown of ABA.
Trends in Plant Science 4, 472–478.
Cutler, S.R., Rodriguez, P.L., Finkelstein, R.R., and Abrams, S.R. 2010. Abscisic
acid: Emergence of a core signaling network. Annual Review of Plant
Biology 61, 651–679.
Daszkowska-Golec, A., and Szarejko, I. 2013. Open or close the gate—­
Stomata action under the control of phytohormones in drought stress
conditions. Frontiers in Plant Science 4, 138.
Davies, W., Kudoyarova, G., and Hartung, W. 2005. Long-distance ABA sig-
naling and its relation to other signaling pathways in the detection of
soil drying and the mediation of the plant’s response to drought. Journal
of Plant Growth Regulation 24, 285–295.
Desikan, R., Hancock, J.T., Bright, J., Harrison, J., Weir, I., Hooley, R., and
Neill, S.J. 2005. A role for ETR1 in hydrogen peroxide signaling in sto-
matal guard cells. Plant Physiology 137, 831–834.
Desikan, R., Last, K., Harrett-Williams, R., Tagliavia, C., Harter, K., Hooley,
R., Hancock, J.T., and Neill, S.J. 2006. Ethylene-induced stomatal clo-
sure in Arabidopsis occurs via AtrbohF-mediated hydrogen peroxide
synthesis. Plant Journal 47, 907–916.
Dietz, K.J., Sauter, A., Wichert, K., Messdaghi, D., and Hartung, W. 2000.
Extracellular β-glucosidase activity in barley involved in the hydrolysis
of ABA glucose conjugate in leaves. Journal of Experimental Botany 51,
937–944.
Dodd, A.N., Salathia, N., Hall, A., Kévei, E., Tóth, R., Nagy, F., Hibberd, J.M.,
Millar, A.J., and Webb, A.A.R. 2005. Plant circadian clocks increase pho-
tosynthesis, growth, survival, and competitive advantage. Science 309,
630–633.

195
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Doheny-Adams, T., Hunt, L., Franks, P.J., Beerling, D.J., and Gray, J.E. 2012.
Genetic manipulation of stomatal density influences stomatal size, plant
growth and tolerance to restricted water supply across a growth car-
bon dioxide gradient. Philosophical Transactions of the Royal Society B:
Biological Sciences 367, 547–555.
Dong, X., Ramírez-Sánchez, M., Huber, D.J., Rao, J., Zhang, Z., Choi, S.T.,
and Lee, J.H. 2013. Diffusivity of 1-methylcyclopropene in spinach and
bok choi leaf tissue, disks of tomato and avocado fruit tissue, and whole
tomato fruit. Postharvest Biology and Technology 78, 40–47.
Drake, P.L., Froend, R.H., and Franks, P.J. 2013. Smaller, faster stomata: scal-
ing of stomatal size, rate of response, and stomatal conductance. Journal
of Experimental Botany 64, 495–505.
Dubovskaya, L.V., Bakakina, Y.S., Kolesneva, E.V., Sodel, D.L., McAinsh,
M.R., Hetherington, A.M., and Volotovski, I.D. 2011. cGMP-depen-
dent ABA-induced stomatal closure in the ABA-insensitive Arabidopsis
mutant abi1-1. New Phytologist 191, 57–69.
Effmert, U., Grosse, J., Rose, U.S.R., Ehrig, F., Kagi, R., and Piechulla, B.
2005. Volatile composition, emission pattern, and localization of floral
scent emission in Mirabilis jalapa (Nyctaginaceae). American Journal
of Botany 92, 2–12.
Elibox, W., and Umaharan, P. 2008. Morphophysiological characteristics
associated with vase life of cut flowers of anthurium. HortScience 43,
825–831.
Elibox, W., and Umaharan, P. 2010. Cultivar differences in the deterioration
of vase-life in cut-flowers of Anthurium andraeanum is determined by
mechanisms that regulate water uptake. Scientia Horticulturae 124,
102–108.
Elzenga, J.T.M., and Van Volkenburgh, E. 1997. Kinetics of Ca2+- and ATP-
dependent, voltage-controlled anion conductance in the plasma mem-
brane of mesophyll cells of Pisum sativum. Planta 201, 415–423.
Elzenga, J.T.M., Staal, M., and Prins, H.B.A. 2000. Modulation by phyto-
chrome of the blue light-induced extracellular acidification by leaf epi-
dermal cells of pea (Pisum sativum L.): a kinetic analysis. Plant Journal
22, 377–389.
Endo, A., Sawada, Y., Takahashi, H., Okamoto, M., Ikegami, K., Koiwai, H.,
Seo, M., Toyomasu, T., Mitsuhashi, W., Shinozaki, K., Nakazono, M.,
Kamiya, Y., Koshiba, T., and Nambara, T. 2008. Drought induction of
arabidopsis 9-cis-epoxycarotenoid dioxygenase occurs in vascular
parenchyma cells. Plant Physiology 147, 1984–1993.
Ezell, B.D., and Wilcox, M.S. 1959. Vegetable vitamins, loss of vitamin C
in fresh vegetables as related to wilting and temperature. Journal of
Agricultural and Food Chemistry 7, 507–509.
Ezell, B.B., and Wilcox, M.S. 1962. Vegetable vitamin values, loss of caro-
tene in fresh vegetables as related to wilting and temperature. Journal
of Agricultural and Food Chemistry 10, 124–126.

196
 Stom ata a n d P ostharvest P hy s i o l o g y

Fallik, E., Grinberg, S., Alkalai, S., Yekutieli, O., Wiseblum, A., Regev, R.,
Beres, H., and Bar-Lev, E. 1999. A unique rapid hot water treatment
to improve storage quality of sweet pepper. Postharvest Biology and
Technology 15, 25–32.
Fanourakis, D., Carvalho, S.M.P., Almeida, D.P.F., and Heuvelink, E. 2011.
Avoiding high relative air humidity during critical stages of leaf ontogeny
is decisive for stomatal functioning. Physiologia Plantarum 142, 274–286.
Fanourakis, D., Heuvelink, E., and Carvalho, S.M.P. 2013a. A comprehensive
analysis of the physiological and anatomical components involved in
higher water loss rates after leaf development at high humidity. Journal
of Plant Physiology 170, 890–898.
Fanourakis, D., Pieruschka, R., Savvides, A., Macnish, A.J., Sarlikioti, V.,
and Woltering, E.J. 2013b. Sources of vase life variation in cut roses: a
review. Postharvest Biology and Technology 78, 1–15.
Feller, U. 2006. Stomatal opening at elevated temperature: an underestimated
regulatory mechanism? Genaral and Applied Plant Physiology 32, 19–31.
Ferrante, A., Vernieri, P., Serra, G., and Tognoni, F. 2004. Changes in abscisic
acid during leaf yellowing of cut stock flowers. Plant Growth Regulation
43, 127–134.
Fonseca, J.M., Kim, H.J., Kline, W.L., Wyenandt, C.A., Hoque, M., Ajwa,
H., and French, N. 2009. Effect of preharvest application of a second-
generation harpin protein on microbial quality, antioxidants, and shelf
life of fresh-cut lettuce. Journal of the American Society for Horticultural
Science 134, 141–147.
Fordham, M.C., Harrison-Murray, R.S., Knight, L., and Evered, C.E. 2001. Effects
of leaf wetting and high humidity on stomatal function in leafy cuttings and
intact plants of Corylus maxima. Physiologia Plantarum 113, 233–240.
Franck, C., Lammertyn, J., Ho, Q.T., Verboven, P., Verlinden, B., and Nicolaï,
B.M. 2007. Browning disorders in pear fruit. Postharvest Biology and
Technology 43, 1–13.
Franks, P.J., and Farquhar, G.D. 2007. The mechanical diversity of stomata
and its significance in gas-exchange control. Plant Physiology 143, 78–87.
Frechilla, S., Zhu, J., Talbott, L.D., and Zeiger, E. 1999. Stomata from npq1,
a zeaxanthin-less Arabidopsis mutant, lack a specific response to blue
light. Plant and Cell Physiology 40, 949–954.
Frommhold, I. 1982. Effect of ethephon on stomatal opening in detached
epidermal strips of tobacco leaves (Nicotiana tobacum L. cv. Samsun).
Biologia Plantarum 24, 303–306.
Fujii, H., and Zhu, J.K. 2009. Arabidopsis mutant deficient in 3 abscisic acid-
activated protein kinases reveals critical roles in growth, reproduction,
and stress. Proceedings of the National Academy of Sciences of the United
States of America 106, 8380–8385.
Fujii, H., Chinnusamy, V., Rodrigues, A., Rubio, S., Antoni, R., Park, S.Y.,
Cutler, S.R., Sheen, J., Rodriguez, P.L., and Zhu, J.K. 2009. In vitro recon-
stitution of an abscisic acid signaling pathway. Nature 462, 660–664.

197
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Fujita, Y., Nakashima, K., Yoshida, T., Katagiri, T., Kidokoro, S., Kanamori,
N., Umezawa, T., Fujita, M., Maruyama, K., Ishiyama, K., Kobayashi,
M., Nakasone, S., Yamada, K., Ito, T., Shinozaki, K., and Yamaguchi-
Shinozaki, K. 2009. Three SnRK2 protein kinases are the main posi-
tive regulators of abscisic acid signaling in response to water stress in
Arabidopsis. Plant and Cell Physiology 50, 2123–2132.
Gan, S., and Amasino, R.M. 1995. Inhibition of leaf senescence by autoregu-
lated production of cytokinin. Science 270, 1986–1988.
Geiger, D., Scherzer, S., Mumm, P., Marten, I., Ache, P., Matschi, S., Liese, A.,
Wellmann, C., Al-Rasheid, K.A.S., Grill, E. et al. 2010. Guard cell anion
channel SLAC1 is regulated by CDPK protein kinases with distinct Ca2+
affinities. Proceedings of the National Academy of Sciences of the United
States of America 107, 8023–8028.
Geiger, D., Scherzer, S., Mumm, P., Stange, A., Marten, I., Bauer, H., Ache,
P., Matschi, S., Liese, A., Al-Rasheid, K.A.S., Grill, E., Romeis, T., and
Hedrich, R. 2009. Activity of guard cell anion channel SLAC1 is con-
trolled by drought-stress signaling kinase-phosphatase pair. Proceedings
of the National Academy of Sciences of the United States of America 106,
21425–21430.
Ghashghaie, J., Brenckmann, F., and Saugier, B. 1992. Water relations and
growth of rose plants cultured in vitro under various relative humidities.
Plant Cell, Tissue and Organ Culture 30, 51–57.
Giday, H., Fanourakis, D., Kjaer, K.H., Fomsgaard, I.S., and Ottosen, C.O.
2013a. Foliar abscisic acid content underlies genotypic variation in sto-
matal responsiveness after growth at high relative air humidity. Annals
of Botany 112, 1857–1867.
Giday, H., Kjaer, K.H., Fanourakis, D., and Ottosen, C.O. 2013b. Smaller
stomata require less severe leaf drying to close: A case study in Rosa
hydrida. Journal of Plant Physiology 170, 1309–1316.
Giday, H., Fanourakis, D., Kjaer, K.H., Fomsgaard, I.S., and Ottosen, C.O.
2014. Threshold response of stomatal closing ability to leaf abscisic
acid concentration during growth. Journal of Experimental Botany, doi:
10.1093/jxb/eru216.
Gudesblat, G.E., Torres, P.S., and Vojnov, A.A. 2009. Stomata and pathogens:
Warfare at the gates. Plant Signaling and Behavior 4, 1114–1116.
Guo, Y., Xiong, L., Song, C.P., Gong, D., Halfter, U., and Zhu, J.K. 2002. A
calcium sensor and its interacting protein kinase are global regulators
of abscisic acid signaling in arabidopsis. Developmental Cell 3, 233–244.
Halevy, A.H., and Mayak, S. 1981. Senescence and postharvest physiology of
cut flowers. Part 2. Horticultural Reviews 3, 59–143.
Halevy, A.H., Mayak, S., Tirosh, T., Spiegelstein, H., and Kofranek, A.M.
1974. Opposing effects of abscisic acid on senescence of rose flowers.
Plant and Cell Physiology 15, 813–821.
Hall, A.E., Camacho, B.S.E., and Kaufmann, M.R. 1975. Regulation of water
loss by citrus leaves. Physiologia Plantarum 33, 62–65.

198
 Stom ata a n d P ostharvest P hy s i o l o g y

Hao, J.H., Wang, X.L., Dong, C.J., Zhang, Z.G., and Shang, Q.M. 2011.
Salicylic acid induces stomatal closure by modulating endogenous hor-
mone levels in cucumber cotyledons. Russian Journal of Plant Physiology
58, 906–913.
Harada, A., and Shimazaki, K.I. 2009. Measurement of changes in cytosolic
Ca2+ in Arabidopsis guard cells and mesophyll cells in response to blue
light. Plant and Cell Physiology 50, 360–373.
Hauser, F., Waadt, R., and Schroeder, J.I. 2011. Evolution of abscisic acid syn-
thesis and signaling mechanisms. Current Biology 21, R346–R355.
Hazarika, B.N. 2006. Morpho-physiological disorders in in vitro culture of
plants. Scientia Horticulturae 108, 105–120.
Hentzen, A.E., Smart, L.B., Wimmers, L.E., Fang, H.H., Schroeder, J.I.,
and Bennett, A.B. 1996. Two plasma membrane H+ -ATPase genes
expressed in guard cells of Vicia faba are also expressed throughout
the plant. Plant and Cell Physiology 37, 650–659.
Herppich, W.B., Linke, M., Landahl, S., and Gzik, A. 2001. Preharvest and
postharvest responses of radish to reduced water supply during growth.
Acta Horticulturae 553, 89–90.
Hetherington, A.M., and Woodward, F.I. 2003. The role of stomata in sensing
and driving environmental change. Nature 424, 901–908.
Hew, C.S., Lee, G.L., and Wong, S.C. 1980. Occurrence of non-functional sto-
mata in the flowers of tropical orchids. Annals of Botany 46, 195–201.
Hewett, E.W. 2006. An overview of preharvest factors influencing postharvest
quality of horticultural products. International Journal of Postharvest
Technology and Innovation 1, 4–15.
Holbrook, N.M., Shashidhar, V., James, R.A., and Munns, R. 2002. Stomatal
control in tomato with ABA‐deficient roots: Response of grafted plants
to soil drying. Journal of Experimental Botany 53, 1503–1514.
Hossain, M.A., Munemasa, S., Uraji, M., Nakamura, Y., Mori, I.C., and Murata,
Y. 2011. Involvement of endogenous abscisic acid in methyl jasmonate-
induced stomatal closure in Arabidopsis. Plant Physiology 156, 430–438.
Hronková, M., Zahradníčková, H., Šimková, M., Šimek, P., and Heydová, A.
2003. The role of abscisic acid in acclimation of plants cultivated in vitro
to ex vitro conditions. Biologia Plantarum 46, 535–541.
Hu, X., Zhang, A., Zhang, J., and Jiang, M. 2006. Abscisic acid is a key inducer
of hydrogen peroxide production in leaves of maize plants exposed to
water stress. Plant and Cell Physiology 47, 1484–1495.
Huang, X.Y., Chao, D.Y., Gao, J.P., Zhu, M.Z., Shi, M., and Lin, H.X. 2009. A
previously unknown zinc finger protein, DST, regulates drought and salt
tolerance in rice via stomatal aperture control. Genes and Development
23, 1805–1817.
Hubbard, K.E., Nishimura, N., Hitomi, K., Getzoff, E.D., and Schroeder,
J.I. 2010. Early abscisic acid signal transduction mechanisms: Newly
discovered components and newly emerging questions. Genes and
Development 24, 1695–1708.

199
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Hubbard, K.E., Siegel, R.S., Valerio, G., Brandt, B., and Schroeder, J.I. 2012.
Abscisic acid and CO2 signaling via calcium sensitivity priming in guard
cells, new CDPK mutant phenotypes and a method for improved resolu-
tion of stomatal stimulus–response analyses. Annals of Botany 109, 5–17.
Hunter, D.A., Ferrante, A., Vernieri, P., and Reid, M.S. 2004. Role of abscisic
acid in perianth senescence of daffodil (Narcissus pseudonarcissus
‘Dutch Master’). Physiologia Plantarum 121, 313–321.
In, B.C., Motomura, S., Inamoto, K., Doi, M., and Mori, G. 2007. Multivariate
analysis of relations between preharvest environmental factors, post-
harvest morphological and physiological factors, and vase life of cut
‘Asami Red’ roses. Journal of the Japanese Society for Horticultural
Science 76, 66–72.
Islam, N., Torre, S., Wold, A.B., and Gislerød, H.R. 2010. Effects of growing
conditions on the postharvest quality of herbs. Acta Horticulturae 877,
187–194.
Iuchi, S., Kobayashi, M., Taji, T., Naramoto, M., Seki, M., Kato, T., Tabata,
S., Kakubari, Y., Yamaguchi‐Shinozaki, K., and Shinozaki, K. 2001.
Regulation of drought tolerance by gene manipulation of 9‐cis‐epoxy-
carotenoid dioxygenase, a key enzyme in abscisic acid biosynthesis in
Arabidopsis. Plant Journal 27, 325–333.
Jiang, F., and Hartung, W. 2008. Long-distance signaling of abscisic acid
(ABA): the factors regulating the intensity of the ABA signal. Journal of
Experimental Botany 59, 37–43.
Johnson, B.E., and Brun, W. 1966. Stomatal density and responsiveness of
banana fruit stomates. Plant Physiology 41, 99–101.
Jones, H.G. 1999. Use of thermography for quantitative studies of spatial and
temporal variation of stomatal conductance over leaf surfaces. Plant,
Cell and Environment 22, 1043–1055.
Joshi-Saha, A., Valon, C., and Leung, J. 2011. Abscisic acid signal off the
STARTing block. Molecular Plant 4, 562–580.
Khan, K., Joshi, P., and Purohit, S.D. 2010. Stomatal characteristics dur-
ing micropropagation of Wrightia tomentosa. Acta Horticulturae 865,
187–192.
Khokon, M.A.R., Okuma, E., Hossain, M.A., Munemasa, S., Uraji, M.,
Nakamura, Y., Mori, I.C., and Murata, Y. 2011. Involvement of extra-
cellular oxidative burst in salicylic acid-induced stomatal closure in
Arabidopsis. Plant, Cell and Environment 34, 434–443.
Kim, T.H., Böhmer, M., Hu, H., Nishimura, N., and Schroeder, J.I. 2010.
Guard cell signal transduction network: advances in understanding
abscisic acid, CO2, and Ca2+ signaling. Annual Review of Plant Biology
61, 561–591.
Kirk, H., and Skytt Anderson, A. 1985. Influence of low pressure storage on sto-
matal opening and rooting of cuttings. Acta Horticulturae 181, 393–398.
Kirk, H., Andersen, A.S., Veierskov, B., Johansen, E., and Aabrandt, Z. 1986.
Low-pressure storage of hibiscus cuttings. Effect on stomatal opening
and rooting. Annals of Botany 58, 389–396.

200
 Stom ata a n d P ostharvest P hy s i o l o g y

Koiwai, H., Nakaminami, K., Seo, M., Mitsuhashi, W., Toyomasu, T., and
Koshiba, T. 2004. Tissue-specific localization of an abscisic acid biosyn-
thetic enzyme, AAO3, in arabidopsis. Plant Physiology 134, 1697–1707.
Krochko, J.E., Abrams, G.D., Loewen, M.K., Abrams, S.R., and Cutler, A.J.
1998. (+)-Abscisic acid 8´-hydroxylase is a cytochrome P450 monooxy-
genase. Plant Physiology 118, 849–860.
Kroupitski, Y., Golberg, D., Belausov, E., Pinto, R., Swartzberg, D., Granot,
D., and Sela, S. 2009. Internalization of Salmonella enterica in leaves is
induced by light and involves chemotaxis and penetration through open
stomata. Applied and Environmental Microbiology 75, 6076–6086.
Kuromori, T., Sugimoto, E., and Shinozaki, K. 2014. Intertissue signal trans-
fer of abscisic acid from vascular cells to guard cells. Plant Physiology
164, 1587–1592.
Kushiro, T., Okamoto, M., Nakabayashi, K., Yamagishi, K., Kitamura, S.,
Asami, T., Hirai, N., Koshiba, T., Kamiya, Y., and Nambara, E. 2004. The
Arabidopsis cytochrome P450 CYP707A encodes ABA 8´-hydroxylases:
key enzymes in ABA catabolism. EMBO Journal 23, 1647–1656.
Kwak, J.M., Moon, J.H., Murata, Y., Kuchitsu, K., Leonhardt, N., DeLong, A.,
and Schroeder, J.I. 2002. Disruption of a guard cell–expressed protein
phosphatase 2A regulatory subunit, RCN1, confers abscisic acid insen-
sitivity in Arabidopsis. Plant Cell 14, 2849–2861.
Laurin, É., Nunes, M.C.N., Émond, J.P., and Brecht, J.K. 2006. Residual effect
of low-pressure stress during simulated air transport on Beit Alpha-type
cucumbers: Stomata behavior. Postharvest Biology and Technology 41,
121–127.
Lazan, H., Ali, Z.M., Mohd, A.A., and Nahar, F. 1987a. Water stress and qual-
ity decline during storage of tropical leafy vegetables. Journal of Food
Science 52, 1286–1292.
Lazan, H., Ali, Z.M., Mohd, A.A., and Ong, G.B. 1987b. Influence of water
stress on cold-induced sweetening in leafy vegetable, Brassica juncea L.
Journal of Food Science 52, 1289–1292.
Lebaudy, A., Vavasseur, A., Hosy, E., Dreyer, I., Leonhardt, N., Thibaud, J.B.,
Véry, A.A., Simonneau, T., and Sentenac, H. 2008. Plant adaptation to
fluctuating environment and biomass production are strongly depen-
dent on guard cell potassium channels. Proceedings of the National
Academy of Sciences of the United States of America 105, 5271–5276.
Leckie, C.P., McAinsh, M.R., Montgomery, L., Priestley, A.J., Staxen, I.,
Webb, A.A.R., and Hetherington, A.M. 1998. Second messengers in
guard cells. Journal of Experimental Botany 49, 339–349.
Lee, K.H., Piao, H.L., Kim, H.Y., Choi, S.M., Jiang, F., Hartung, W., Hwang,
I., Kwak, J.M., and Lee, I.J. 2006. Activation of glucosidase via stress-
induced polymerization rapidly increases active pools of abscisic acid.
Cell 126, 1109–1120.
Lee, S.C., and Luan, S. 2012. ABA signal transduction at the crossroad of
biotic and abiotic stress responses. Plant, Cell and Environment 35,
53–60.

201
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lee, S.C., Lan, W., Buchanan, B.B., and Luan, S. 2009. A protein kinase-phos-
phatase pair interacts with an ion channel to regulate ABA signaling in
plant guard cells. Proceedings of the National Academy of Sciences of the
United States of America 106, 21419–21424.
Lee, S.C., Lim, C.W., Lan, W., He, K., and Luan, S. 2013. ABA signaling in
guard cells entails a dynamic protein-protein interaction relay from the
PYL-RCAR family receptors to ion channels. Molecular Plant 6, 528–538.
Lee, S.K., and Kader, A.A. 2000. Preharvest and postharvest factors influenc-
ing vitamin C content of horticultural crops. Postharvest Biology and
Technology 20, 207–220.
Leonhardt, N., Vavasseur, A., and Forestier, C. 1999. ATP binding cassette
modulators control abscisic acid-regulated slow anion channels in
guard cells. Plant Cell 11, 1141–1151.
Leung, J., Merlot, S., and Giraudat, J. 1997. The Arabidopsis ABSCISIC ACID-
INSENSITIVE2 (ABI2) and ABI1 genes encode homologous protein
phosphatases 2C involved in abscisic acid signal transduction. Plant
Cell 9, 759–771.
Levchenko, V., Konrad, K.R., Dietrich, P., Roelfsema, M.R.G., and Hedrich, R.
2005. Cytosolic abscisic acid activates guard cell anion channels without
preceding Ca2+ signals. Proceedings of the National Academy of Sciences
of the United States of America 102, 4203–4208.
Li, J., and Assmann, S.M. 1996. An abscisic acid-activated and calcium-indepen-
dent protein kinase from guard cells of fava bean. Plant Cell 8, 2359–2368.
Li, S., Assmann, S.M., and Albert, R. 2006. Predicting essential components
of signal transduction networks: a dynamic model of guard cell abscisic
acid signaling. PLoS Biology 4, 1732–1748.
Li, X., Zhu, X., Zhao, N., Fu, D., Li, J., Chen, W., and Chen, W. 2013. Effects
of hot water treatment on anthracnose disease in papaya fruit and its
possible mechanism. Postharvest Biology and Technology 86, 437–446.
López-Gálvez, F., Gil, M.I., Truchado, P., Selma, M.V., and Allende, A. 2010.
Cross-contamination of fresh-cut lettuce after a short-term exposure
during pre-washing cannot be controlled after subsequent washing with
chlorine dioxide or sodium hypochlorite. Food Microbiology 27, 199–204.
Lu, Y.Y., and Chen, C.Y. 2005. Molecular analysis of lily leaves in response to
salicylic acid effective towards protection against Botrytis elliptica. Plant
Science 169, 1–9.
Ma, Y., Szostkiewicz, I., Korte, A., Moes, D., Yang, Y., Christmann, A., and
Grill, E. 2009. Regulators of PP2C phosphatase activity function as
abscisic acid sensors. Science 324, 1064–1068.
MacRobbie, E.A.C. 1990. Calcium-dependent and calcium-independent
events in the initiation of stomatal closure by abscisic acid. Proceedings
of the Royal Society B: Biological Sciences 241, 214–219.
Madhavan, S., Chrominiski, A., and Smith, B.N. 1983. Effect of ethylene
on stomatal opening in tomato and carnation leaves. Plant and Cell
Physiology 24, 569–572.

202
 Stom ata a n d P ostharvest P hy s i o l o g y

Magwaza, L.S., Opara, U.L., Cronje, P.J.R., Landahl, S., and Terry, L.A. 2013.
Canopy position affects rind biochemical profile of ‘Nules Clementine’
mandarin fruit during postharvest storage. Postharvest Biology and
Technology 86, 300–308.
Marten, H., Hedrich, R., and Roelfsema, M.R.G. 2007a. Blue light inhibits
guard cell plasma membrane anion channels in a phototropin-depen-
dent manner. Plant Journal 50, 29–39.
Marten, H., Konrad, K.R., Dietrich, P., Roelfsema, M.R.G., and Hedrich, R.
2007b. Ca2+-dependent and -independent abscisic acid activation of
plasma membrane anion channels in guard cells of Nicotiana tabacum.
Plant Physiology 143, 28–37.
Martin, E.S., and Meidner, H. 1971. Endogenous stomatal movements in
Tradescantia virginiana. New Phytologist 70, 923–928.
Martínez-Sánchez, A., Tudela, J.A., Luna, C., Allende, A., and Gil, M.I. 2011.
Low oxygen levels and light exposure affect quality of fresh-cut Romaine
lettuce. Postharvest Biology and Technology 59, 34–42.
Mayak, S., and Dilley, D. 1976. Effect of sucrose on response of cut carnation
(Dianthus caryophyllus) to kinetin, ethylene, and abscisic acid. Journal
of the American Society for Horticultural Science 101, 583–585.
Mayak, S., and Halevy, A.H. 1974. The action of kinetin in improving the
water balance and delaying sensecence processes of cut rose flowers.
Physiologia Plantarum 32, 330–336.
Mayak, S., Halevy, A., and Katz, M. 1972. Correlative changes in phytohor-
mones in relation to senescence processes in rose petals. Physiologia
Plantarum 27, 1–4.
Melhorn, V., Matsumi, K., Koiwai, H., Ikegami, K., Okamoto, M., Nambara,
E., Bittner, F., and Koshiba, T. 2008. Transient expression of AtNCED3
and AAO3 genes in guard cells causes stomatal closure in Vicia faba.
Journal of Plant Research 121, 125–131.
Melotto, M., Underwood, W., Koczan, J., Nomura, K., and He, S.Y. 2006. Plant
stomata function in innate immunity against bacterial invasion. Cell 126,
969–980.
Melotto, M., Underwood, W., and Sheng, Y.H. 2008. Role of stomata in
plant innate immunity and foliar bacterial diseases. Annual Review of
Phytopathology 46, 101–122.
Mencarelli, F., and Bellincontro, A. 2013. Technology and management of
postharvest dehydration. In Sweet, Reinforced and Fortified Wines:
Grape Biochemistry, Technology and Vinification, ed. F. Mencarelli and
P. Tonutti, 51–75. John Wiley & Son, Chichester, West Sussex, UK.
Mendgen, K., Hahn, M., and Deising, H. 1996. Morphogenesis and mech-
anisms of penetration by plant pathogenic fungi. Annual Review of
Phytopathology 34, 367–386.
Mengel, K., and Kirkby, E.A. 1982. Principles of Plant Nutrition, 655.
International Potash Institute, Bern.

203
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Merlot, S., Leonhardt, N., Fenzi, F., Valon, C., Costa, M., Piette, L., Vavasseur,
A., Genty, B., Boivin, K., Muller, A., Giraudat, J., and Leung, J. 2007.
Constitutive activation of a plasma membrane H+ -ATPase prevents
abscisic acid-mediated stomatal closure. EMBO Journal 26, 3216–3226.
Merlot, S., Mustilli, A.C., Genty, B., North, H., Lefebvre, V., Sotta, B.,
Vavasseur, A., and Giraudat, J. 2002. Use of infrared thermal imaging
to isolate Arabidopsis mutants defective in stomatal regulation. Plant
Journal 30, 601–609.
Merritt, F., Kemper, A., and Tallman, G. 2001. Inhibitors of ethylene synthe-
sis inhibit auxin-induced stomatal opening in epidermis detached from
leaves of Vicia faba L. Plant and Cell Physiology 42, 223–230.
Milillo, S.R., Badamo, J.M., Boor, K.J., and Wiedmann, M. 2008. Growth
and persistence of Listeria monocytogenes isolates on the plant model
Arabidopsis thaliana. Food Microbiology 25, 698–704.
Mintz-Oron, S., Mandel, T., Rogachev, I., Feldberg, L., Lotan, O., Yativ, M.,
Wang, Z., Jetter, R., Venger, I., and Adato, A. 2008. Gene expression
and metabolism in tomato fruit surface tissues. Plant Physiology 147,
823–851.
Moes, D., Himmelbach, A., Korte, A., Haberer, G., and Grill, E. 2008. Nuclear
localization of the mutant protein phosphatase abi1 is required for insen-
sitivity towards ABA responses in Arabidopsis. Plant Journal 54, 806–819.
Montillet, J.L., Leonhardt, N., Mondy, S., Tranchimand, S., Rumeau, D.,
Boudsocq, M., Garcia, A.V., Douki, T., Bigeard, J., Laurière, C.,
Chevalier, A., Castresana, C., and Hirt, H. 2013. An abscisic acid-
independent oxylipin pathway controls stomatal closure and immune
defense in Arabidopsis. PLoS Biology 11, e1001513. doi: 10.1371/journal.
pbio.1001513.
Moran, R.E., DeEll, J.R., and Halteman, W. 2009. Effects of preharvest pre-
cipitation, air temperature, and humidity on the occurrence of soft scald
in ‘Honeycrisp’ apples. HortScience 44, 1645–1647.
Moreshet, S., and Green, G. 1980. Photosynthesis and diffusion conduc-
tance of the Valencia orange fruit under field conditions. Journal of
Experimental Botany 31, 15–27.
Mori, I.C., Murata, Y., Yang, Y., Munemasa, S., Wang, Y.F., Andreoli, S., Tiriac,
H., Alonso, J.M., Harper, J.F., Ecker, J.R., Kwak, J.R., and Schroeder, J.I.
2006. CDPKs CPK6 and CPK3 function in ABA regulation of guard cell
S-type anion- and Ca2+-permeable channels and stomatal closure. PLoS
Biology 4, 1749–1762.
Morison, J.I., and Gifford, R.M. 1983. Stomatal sensitivity to carbon dioxide
and humidity: A comparison of two C3 and two C4 grass species. Plant
Physiology 71, 789–796.
Mortensen, L.M., and Fjeld, T. 1998. Effects of air humidity, lighting period
and lamp type on growth and vase life of roses. Scientia Horticulturae
73, 229–237.

204
 Stom ata a n d P ostharvest P hy s i o l o g y

Mortensen, L.M., and Gislerød, H.R. 1999. Influence of air humidity and light-
ing period on growth, vase life and water relations of 14 rose cultivars.
Scientia Horticulturae 82, 289–298.
Mortensen, L.M., and Gislerod, H.R. 2000. Effect of air humidity on growth,
keeping quality, water relations, and nutrient content of cut roses.
Gartenbauwissenschaft 65, 40–44.
Mortensen, L.M., and Gislerød, H.R. 2011. Vase life: the influence of variation
in air humidity, temperature and super-elevated CO2 concentration in
roses grown under continuous light. European Journal of Horticultural
Science 76, 63–68.
Mulas, M., and Schirra, M. 2007. The effect of heat conditioning treatments
on the postharvest quality of horticultural crops. Stewart Postharvest
Review 3.
Murata, Y., and Mori, I.C. 2013. Stomatal regulation of plant water status. In
Plant Abiotic Stress, ed. M.A. Jenks and P.M. Hasegawa, 47–67. John
Wiley & Son, West Sussex, UK.
Nambara, E., and Marion-Poll, A. 2005. Abscisic acid biosynthesis and catabo-
lism. Annual Review of Plant Biology 56, 165–185.
Negi, J., Matsuda, O., Nagasawa, T., Oba, Y., Takahashi, H., Kawai-Yamada,
M., Uchimiya, H., Hashimoto, M., and Iba, K. 2008. CO2 regulator
SLAC1 and its homologues are essential for anion homeostasis in plant
cells. Nature 452, 483–486.
Neill, S., Barros, R., Bright, J., Desikan, R., Hancock, J., Harrison, J., Morris,
P., Ribeiro, D., and Wilson, I. 2008. Nitric oxide, stomatal closure, and
abiotic stress. Journal of Experimental Botany 59, 165–176.
Njombolwana, N.S., Erasmus, A., van Zyl, J.G., du Plooy, W., Cronje, P.J.R.,
and Fourie, P.H. 2013. Effects of citrus wax coating and brush type on
imazalil residue loading, green mould control and fruit quality retention
of sweet oranges. Postharvest Biology and Technology 86, 362–371.
Nobel, P.S. 1999. Physicochemical and Environmental Plant Physiology. 2nd
ed. Academic Press, San Diego.
Noichinda, S., Bodhipadma, K., Mahamontri, C., Narongruk, T., and Ketsa, S.
2007. Light during storage prevents loss of ascorbic acid, and increases
glucose and fructose levels in Chinese kale Brassica oleracea var. albo-
glabra. Postharvest Biology and Technology 44, 312–315.
Novák, V., and Vidovič, J. 2003. Transpiration and nutrient uptake dynamics
in maize (Zea mays L.). Ecological Modelling 166, 99–107.
Nunes, M.C.N., Brecht, J.K., Morais, A.M.M.B., and Sargent, S.A. 1998.
Controlling temperature and water loss to maintain ascorbic acid levels
in strawberries during postharvest handling. Journal of Food Science 63,
1033–1036.
Oh, M.M., Trick, H.N., and Rajashekar, C.B. 2009. Secondary metabolism
and antioxidants are involved in environmental adaptation and stress
tolerance in lettuce. Journal of Plant Physiology 166, 180–191.

205
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Okamoto, M., Tanaka, Y., Abrams, S.R., Kamiya, Y., Seki, M., and Nambara,
E. 2009. High humidity induces abscisic acid 8´-hydroxylase in stomata
and vasculature to regulate local and systemic abscisic acid responses
in Arabidopsis. Plant Physiology 149, 825–834.
Orsini, F., Alnayef, M., Bona, S., Maggio, A., and Gianquinto, G. 2012. Low
stomatal density and reduced transpiration facilitate strawberry adapta-
tion to salinity. Environmental and Experimental Botany 81, 1–10.
Osakabe, Y., Yamaguchi-Shinozaki, K., Shinozaki, K., and Tran, L.S.P. 2013.
ABA control of plant macroelement membrane transport systems in
response to water deficit and high salinity. New Phytologist 202, 35–49.
Outlaw, W.H. 2003. Integration of cellular and physiological functions of
guard cells. Critical Reviews in Plant Sciences 22, 503–529.
Pallaghy, C., and Raschke, K. 1972. No stomatal response to ethylene. Plant
Physiology 49, 275–276.
Pallas, J.E., and Kays, S.J. 1982. Inhibition of photosynthesis by ethylene—
a stomatal effect. Plant Physiology 70, 598–601.
Pandey, R., Chacko, P.M., Choudhary, M.L., Prasad, K.V., and Pal, M.
2007. Higher than optimum temperature under CO2 enrichment influ-
ences stomata anatomical characters in rose (Rosa hybrida). Scientia
Horticulturae 113, 74–81.
Pandey, R., Chacko, P.M., Pal, M., Choudhary, M.L., and Singh, R. 2010.
Influence of growth temperature on keeping quality traits of rose (Rosa
hybrida L.) cut flowers grown under continuous CO2 enrichment. Indian
Journal of Plant Physiology 15, 343–349.
Parés, J., Arizaleta, M., Sanabria, M.E., and García, G. 2008. Effect of salin-
ity levels on the stomatal density, stomatal index and leaf thickness of
Carica papaya L. Acta Botánica Venezuelica 31, 27–34.
Park, S.Y., Fung, P., Nishimura, N., Jensen, D.R., Fujii, H., Zhao, Y., Lumba,
S., Santiago, J., Rodrigues, A., Chow, T.F.F., Alfred, S.E., Bonetta, D.,
Finkelstein, R., Provart, N.J., Desveaux, D., Rodriguez, P.L., McCourt,
P., Zhu, J.K., Schroeder, J.I., Volkman, B.F., and Cutler, S.R. 2009.
Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL
family of START proteins. Science 324, 1068–1071.
Parvathi, K., and Raghavendra, A.S. 1997. Blue light-promoted stomatal open-
ing in abaxial epidermis of Commelina benghalensis is maximal at low
calcium. Physiologia Plantarum 101, 861–864.
Paull, R.E. 1982. Anthurium (Anthurium andraeanum Andre) vase life evalu-
ation criteria. HortScience 17, 606–607.
Paull, R.E. 1999. Effect of temperature and relative humidity on fresh com-
modity quality. Postharvest Biology and Technology 15, 263–277.
Peana, A.T., D’Aquila, P.S., Panin, F., Serra, G., Pippia, P., and Moretti, M.D.L.
2002. Anti-inflammatory activity of linalool and linalyl acetate constitu-
ents of essential oils. Phytomedicine 9, 721–726.

206
 Stom ata a n d P ostharvest P hy s i o l o g y

Peschel, S., Beyer, M., and Knoche, M. 2003. Surface characteristics of sweet
cherry fruit: stomata-number, distribution, functionality and surface
wetting. Scientia Horticulturae 97, 265–278.
Peterson, K.M., Rychel, A.L., and Torii, K.U. 2010. Out of the mouths of
plants: The molecular basis of the evolution and diversity of stomatal
development. Plant Cell 22, 296–306.
Pettersen, R.I., Moe, R., and Gislerød, H.R. 2007. Growth of pot roses and post-
harvest rate of water loss as affected by air humidity and temperature
variations during growth under continuous light. Scientia Horticulturae
114, 207–213.
Pettersen, R.I., Mortensen, L.M., Moe, R., and Gislerød, H.R. 2006. Air humid-
ity control essential for rose production under continuous lighting. Acta
Horticulturae 711, 323–331.
Pompodakis, N.E., Joyce, D.C., Terr y, L.A., and L ydakis, D.E. 2004.
Effects of vase solution pH and abscisic acid on the longevity of cut
‘Baccara’ roses. Journal of Horticultural Science and Biotechnology
79, 828–832.
Porat, R., Daus, A., Weiss, B., Cohen, L., Fallik, E., and Droby, S. 2000.
Reduction of postharvest decay in organic citrus fruit by a short hot
water brushing treatment. Postharvest Biology and Technology 18,
151–157.
Pospíšilová, J., Synková, H., and Rulcová, J. 2000. Cytokinins and water stress.
Biologia Plantarum 43, 321–328.
Priest, D.M., Ambrose, S.J., Vaistij, F.E., Elias, L., Higgins, G.S., Ross, A.R.,
Abrams, S.R., and Bowles, D.J. 2006. Use of the glucosyltransferase
UGT71B6 to disturb abscisic acid homeostasis in Arabidopsis thaliana.
Plant Journal 46, 492–502.
Qin, X., and Zeevaart, J.A. 1999. The 9-cis-epoxycarotenoid cleavage reaction
is the key regulatory step of abscisic acid biosynthesis in water-stressed
bean. Proceedings of the National Academy of Sciences of the United States
of America 96, 15354–15361.
Qin, X., and Zeevaart, J.A.D. 2002. Overexpression of a 9-cis-epoxycarotenoid
dioxygenase gene in Nicotiana plumbaginifolia increases abscisic
acid and phaseic acid levels and enhances drought tolerance. Plant
Physiology 128, 544–551.
Raghavendra, A.S., Gonugunta, V.K., Christmann, A., and Grill, E. 2010. ABA
perception and signaling. Trends in Plant Science 15, 395–401.
Reid, M.S., and Jiang, C.Z. 2012. Postharvest biology and technology of cut
flowers and potted plants. Horticultural Reviews 40, 1–54.
Reynolds-Henne, C.E., Langenegger, A., Mani, J., Schenk, N., Zumsteg, A.,
and Feller, U. 2010. Interactions between temperature, drought and sto-
matal opening in legumes. Environmental and Experimental Botany 68,
37–43.

207
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Rezaei Nejad, A., and van Meeteren, U. 2005. Stomatal response character-
istics of Tradescantia virginiana grown at high relative air humidity.
Physiologia Plantarum 125, 324–332.
Rezaei Nejad, A., and van Meeteren, U. 2007. The role of abscisic acid in
disturbed stomatal response characteristics of Tradescantia virginiana
during growth at high relative air humidity. Journal of Experimental
Botany 58, 627–636.
Rezaei Nejad, A., and van Meeteren, U. 2008. Dynamics of adaptation of stoma-
tal behaviour to moderate or high relative air humidity in Tradescantia
virginiana. Journal of Experimental Botany 59, 289–301.
Rezaei Nejad, A., Harbinson, J., and van Meeteren, U. 2006. Dynamics of spa-
tial heterogeneity of stomatal closure in Tradescantia virginiana altered
by growth at high relative air humidity. Journal of Experimental Botany
57, 3669–3678.
Riederer, M., and Muller, C. 2008. Annual Plant Reviews, Biology of the Plant
Cuticle 23. John Wiley & Son, West Sussex, UK.
Rogers, C.A., Powell, R.D., and Sharpe, P.J.H. 1979. Relationship of tempera-
ture to stomatal aperture and potassium accumulation in guard cells of
Vicia faba. Plant Physiology 63, 388–391.
Rogiers, S.Y., Hatfield, J.M., Jaudzems, V.G., White, R.G., and Keller, M.
2004. Grape berry cv. Shiraz epicuticular wax and transpiration during
ripening and preharvest weight loss. American Journal of Enology and
Viticulture 55, 121–127.
Romero, P., Rodrigo, M.J., and Lafuente, M.T. 2013. Differential expression
of the Citrus sinensis ABA perception system genes during postharvest
fruit dehydration. Postharvest Biology and Technology 76, 65–73.
Romero-Aranda, R., Cantó-Garay, R., and Martínez, P.F. 1994. Distribution
and density of stomata in two cultivars of Gerbera jamesonii and its rela-
tion to leaf conductance. Scientia Horticulturae 58, 167–173.
Roychoudhury, A., Paul, S., and Basu, S. 2013. Cross-talk between abscisic
acid-dependent and abscisic acid-independent pathways during abiotic
stress. Plant Cell Reports 32, 985–1006.
Runkle, E., Woolard, D., Campbell, C., Blanchard, M., and Newton, L. 2007.
Exogenous applications of abscisic acid improved the postharvest
drought tolerance of several annual bedding plants. Acta Horticulturae
755, 127–133.
Rutland, R.B., Chang, H.L.L., and Pallas, J.E., Jr. 1987. Stomatal density of
snapdragon as a possible determinant of transpiration. HortScience 22,
599–604.
Sachs, T. 1991. Stomata as an example of meristmoid development. In Pattern
Formation in Plant Tissues, 101–117. Cambridge University Press,
Cambridge.
Sack, F.D. 1987. The development and structure of stomata. In Stomatal
Function, ed. E. Zeiger, G.D. Farquhar, and I.R. Cowan, 59–89. Stanford
University Press, Stanford, CA.

208
 Stom ata a n d P ostharvest P hy s i o l o g y

Saito, S., Hirai, N., Matsumoto, C., Ohigashi, H., Ohta, D., Sakata, K., and
Mizutani, M. 2004. Arabidopsis CYP707As encode (+)-abscisic acid
8´-hydroxylase, a key enzyme in the oxidative catabolism of abscisic
acid. Plant Physiology 134, 1439–1449.
Sams, C.E. 1999. Preharvest factors affecting postharvest texture. Postharvest
Biology and Technology 15, 249–254.
Santamaria, J.M., and Kerstiens, G. 1994. The lack of control of water loss
in micropropagated plants is not related to poor cuticle development.
Physiologia Plantarum 91, 191–195.
Santamaria, J.M., Davies, W.J., and Atkinson, C.J. 1993. Stomata of microprop-
agated Delphinium plants respond to ABA, CO2, light and water poten-
tial, but fail to close fully. Journal of Experimental Botany 44, 99–107.
Santiago, J., Rodrigues, A., Saez, A., Rubio, S., Antoni, R., Dupeux, F., Park,
S.Y., Márquez, J.A., Cutler, S.R., and Rodriguez, P.L. 2009. Modulation
of drought resistance by the abscisic acid receptor PYL5 through inhibi-
tion of clade A PP2Cs. Plant Journal 60, 575–588.
Sauter, A., Dietz, K.J., and Hartung, W. 2002. A possible stress physiological
role of abscisic acid conjugates in root to shoot signaling. Plant, Cell and
Environment 25, 223–228.
Savvides, A., Fanourakis, D., and van Ieperen, W. 2012. Co-ordination of
hydraulic and stomatal conductances across light qualities in cucumber
leaves. Journal of Experimental Botany 63, 1135–1143.
Schirra, M., D’Hallewin, G., Ben-Yehoshua, S., and Fallik, E. 2000. Host–
pathogen interactions modulated by heat treatment. Postharvest Biology
and Technology 21, 71–85.
Schreiber, U., and Berry, J.A. 1977. Heat-induced changes of chlorophyll fluo-
rescence in intact leaves correlated with damage of the photosynthetic
apparatus. Planta 136, 233–238.
Schroeder, J.I., and Keller, B.U. 1992. Two types of anion channel cur-
rents in guard cells with distinct voltage regulation. Proceedings of
the National Academy of Sciences of the United States of America 89,
5025–5029.
Schroeder, K.R., and Stimart, D.P. 2005. Comparison of stomatal density
and postharvest transpiration between long- and short-lived cut flower
genotypes of Antirrhinum majus L. Journal of the American Society for
Horticultural Science 130, 742–746.
Schroeder, J.I., Allen, G.J., Hugouvieux, V., Kwak, J.M., and Waner, D. 2001.
Guard cell signal transduction. Annual Review of Plant Biology, 52,
627–658.
Schulze, E.D., Lange, O.L., Kappen, L., Buschbom, U., and Evenari, M. 1973.
Stomatal responses to changes in temperature at increasing water
stress. Planta 110, 29–42.
Schwertfeger, G., and Buchloh, G. 1968. Die Korkentwicklung in den
Abschlussgeweben der Triebe und Fruchte verschiedener Apfelsorten.
Gartenbauwissenschaft 33, 77–102.

209
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Shamaila, M. 2005. Water and its relation to fresh produce. In Produce

Degradation: Pathways and Prevention, ed. S.I. Dike Ukuku and
O. Lamikanra, 267–291. CRC Press, Boca Raton, FL.
She, X., and Song, X. 2012. Ethylene inhibits abscisic acid-induced stomatal
closure in Vicia faba via reducing nitric oxide levels in guard cells. New
Zealand Journal of Botany 50, 203–216.
Shimizu-Yumoto, H., and Ichimura, K. 2009. Abscisic acid, in combination with
sucrose, is effective as a pulse treatment to suppress leaf damage and
extend foliage vase-life in cut Eustoma flowers. Journal of Horticultural
Science and Biotechnology 84, 107–111.
Shtein, I., Meir, S., Riov, J., and Philosoph-Hadas, S. 2011. Interconnection
of seasonal temperature, vascular traits, leaf anatomy and hydraulic
performance in cut Dodonaea ‘Dana’ branches. Postharvest Biology and
Technology 61, 184–192.
Siegel, R.S., Xue, S., Murata, Y., Yang, Y., Nishimura, N., Wang, A., and
Schroeder, J.I. 2009. Calcium elevation-dependent and attenuated rest-
ing calcium-dependent abscisic acid induction of stomatal closure and
abscisic acid-induced enhancement of calcium sensitivities of S-type
anion and inward-rectifying K+ channels in Arabidopsis guard cells.
Plant Journal 59, 207–220.
Sirichandra, C., Wasilewska, A., Vlad, F., Valon, C., and Leung, J. 2009. The
guard cell as a single-cell model towards understanding drought tolerance
and abscisic acid action. Journal of Experimental Botany 60, 1439–1463.
Sisler, E.C., and Wood, C. 1988. Interaction of ethylene and CO2. Physiologia
Plantarum 73, 440–444.
Slootweg, G., and van Meeteren, U. 1991. Transpiration and stomatal con-
ductance of roses cv. Sonia grown with supplemental lighting. Acta
Horticulturae, 298, 119–125.
Smith, D.L., Stommel, J.R., Fung, R.W.M., Wang, C.Y., and Whitaker, B.D.
2006. Influence of cultivar and harvest method on postharvest storage
quality of pepper (Capsicum annuum L.) fruit. Postharvest Biology and
Technology 42, 243–247.
Smith, K., Fleming, H., Van Dyke, C., and Lower, R. 1979. Scanning electron
microscopy of the surface of the pickling cucumber fruit. Journal of the
American Society for Horticultural Science 104, 528–533.
Song, C.P., Agarwal, M., Ohta, M., Guo, Y., Halfter, U., Wang, P., and Zhu, J.K.
2005. Role of an Arabidopsis AP2/EREBP-type transcriptional repressor
in abscisic acid and drought stress responses. Plant Cell 17, 2384–2396.
Song, X.G., She, X.P., and Wang, J. 2012. Inhibition of darkness-induced sto-
matal closure by ethylene involves a removal of hydrogen peroxide
from guard cells of Vicia faba. Russian Journal of Plant Physiology 59,
372–380.
Song, X.G., She, X.P., He, J.M., Huang, C., and Song, T.S. 2006. Cytokinin- and
auxin-induced stomatal opening involves a decrease in levels of hydro-
gen peroxide in guard cells of Vicia faba. Functional Plant Biology 33,
573–583.

210
 Stom ata a n d P ostharvest P hy s i o l o g y

Sreenivasulu, N., Harshavardhan, V.T., Govind, G., Seiler, C., and Kohli, A.
2012. Contrapuntal role of ABA: Does it mediate stress tolerance or
plant growth retardation under long-term drought stress? Gene 506,
265–273.
Stael, S., Wurzinger, B., Mair, A., Mehlmer, N., Vothknecht, U.C., and Teige,
M. 2011. Plant organellar calcium signaling: An emerging field. Journal
of Experimental Botany 63, 1525–1542.
Su, J.S., Wang, Y.F., Frelet, A., Leonhardt, N., Klein, M., Forestier, C., Mueller-
Roeber, B., Cho, M.H., Martinoia, E., and Schroeder, J.I. 2007. The
ATP binding cassette transporter AtMRP5 modulates anion and cal-
cium channel activities in Arabidopsis guard cells. Journal of Biological
Chemistry 282, 1916–1924.
Suhita, D., Raghavendra, A.S., Kwak, J.M., and Vavasseur, A. 2004.
Cytoplasmic alkalization precedes reactive oxygen species production
during methyl jasmonate- and abscisic acid-induced stomatal closure.
Plant Physiology 134, 1536–1545.
Sutter, J.U., Sieben, C., Hartel, A., Eisenach, C., Thiel, G., and Blatt, M.R.
2007. Abscisic acid triggers the endocytosis of the Arabidopsis KAT1 K+
channel and its recycling to the plasma membrane. Current Biology 17,
1396–1402.
Swanson, B.T., Jr., Wilkins, H.F., Weiser, C.F., and Klein, I. 1975. Endogenous
ethylene and abscisic acid relative to phytogerontology. Plant Physiology
55, 370–376.
Tahir, S.S., and Rajput, M.T.M. 2010. SEM studies of petal structure of corolla
of the species Sibbaldia L. (Rosaceae). Pakistan Journal of Botany 42,
1443–1449.
Takemiya, A., Kinoshita, T., Asanuma, M., and Shimazaki, K.I. 2006. Protein
phosphatase 1 positively regulates stomatal opening in response to blue
light in Vicia faba. Proceedings of the National Academy of Sciences of the
United States of America 103, 13549–13554.
Talbott, L., and Zeiger, E. 1998. The role of sucrose in guard cell osmoregula-
tion. Journal of Experimental Botany 49, 329–337.
Tallman, G. 2004. Are diurnal patterns of stomatal movement the result of
alternating metabolism of endogenous guard cell ABA and accumula-
tion of ABA delivered to the apoplast around guard cells by transpira-
tion? Journal of Experimental Botany 55, 1963–1976.
Tan, B.C., Joseph, L.M., Deng, W.T., Liu, L., Li, Q.B., Cline, K., and McCarty,
D.R. 2003. Molecular characterization of the Arabidopsis 9-cis-epoxyca-
rotenoid dioxygenase gene family. Plant Journal 35, 44–56.
Tanaka, Y., Sano, T., Tamaoki, M., Nakajima, N., Kondo, N., and Hasezawa,
S. 2005. Ethylene inhibits abscisic acid-induced stomatal closure in
Arabidopsis. Plant Physiology 138, 2337–2343.
Tanaka, Y., Sano, T., Tamaoki, M., Nakajima, N., Kondo, N., and Hasezawa,
S. 2006. Cytokinin and auxin inhibit abscisic acid-induced stomatal
closure by enhancing ethylene production in Arabidopsis. Journal of
Experimental Botany 57, 2259–2266.

211
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Tang, J., Mitcham, E., Wang, S., and Lurie, S. 2007. Heat Treatments for
Postharvest Pest Control: Theory and Practice. CABI, Wallingford, UK.
Taylor, G.E., and Gunderson, C.A. 1986. The response of foliar gas exchange
to exogenously applied ethylene. Plant Physiology 82, 653–657.
Taylor, G.E., and Gunderson, C.A. 1988. Physiological site of ethylene effects
on carbon dioxide assimilation in Glycine max L. Merr. Plant Physiology
86, 85–92.
Tesfay, S.Z., Bertling, I., and Bower, J.P. 2011. Effects of postharvest potassium
silicate application on phenolics and other anti-oxidant systems aligned to
avocado fruit quality. Postharvest Biology and Technology 60, 92–99.
Thimann, K.V. 1985. The interaction of hormonal and environmental factors
in leaf senescence. Biologia Plantarum 27, 83–91.
Thimann, K.V., and Satler, S. 1979. Relation between senescence and stomatal
opening: senescence in darkness. Proceedings of the National Academy
of Sciences of the United States of America 76, 2770–2773.
Thomson, G.E. 2005. Postharvest handling effects on stomatal aperture of
harvested Brassica rapa var. perviridis ‘Komatsuna’ and B. juncea ‘Red
Giant’ mustard leaves. New Zealand Journal of Crop and Horticultural
Science 33, 311–316.
Tibaldi, G., Fontana, E., and Nicola, S. 2011. Growing conditions and posthar-
vest management can affect the essential oil of Origanum vulgare L. ssp.
hirtum (Link) Ietswaart. Industrial Crops and Products 34, 1516–1522.
Tijskens, L.M.M., Veltman, R.H., Heuvelink, E., and Simčič, M. 2003.
Modelling postharvest quality behaviour as affected by preharvest con-
ditions. Acta Horticulturae 599, 469–477.
Tissera, P., and Ayres, P. 1986. Endogenous ethylene affects the behaviour of
stomata in epidermis isolated from rust infected faba bean (Vicia faba
L.). New Phytologist 104, 53–61.
Torre, S. 1999. The role of air humidity and calcium in determining posthar-
vest quality of roses. Institutt for Plantefag, Norges Landbrukshogskole
[Department of Horticulture and Crop Sciences, Agricultural University
of Norway].
Torre, S., and Fjeld, T. 2001. Water loss and postharvest characteristics of
cut roses grown at high or moderate relative air humidity. Scientia
Horticulturae 89, 217–226.
Torre, S., Fjeld, T., and Gislerød, H.R. 2001. Effects of air humidity and K/Ca
ratio in the nutrient supply on growth and postharvest characteristics of
cut roses. Scientia Horticulturae 90, 291–304.
Torre, S., Fjeld, T., Gislerød, H.R., and Moe, R. 2003. Leaf anatomy and sto-
matal morphology of greenhouse roses grown at moderate or high air
humidity. Journal of the American Society for Horticultural Science 128,
598–602.
Tricker, P.J., George Gibbings, J., Rodríguez López, C.M., Hadley, P., and
Wilkinson, M.J. 2012. Low relative humidity triggers RNA-directed de
novo DNA methylation and suppression of genes controlling stomatal
development. Journal of Experimental Botany 63, 3799–3814.

212
 Stom ata a n d P ostharvest P hy s i o l o g y

Trontin, C., Tisné, S., Bach, L., and Loudet, O. 2011. What does Arabidopsis
natural variation teach us (and does not teach us) about adaptation in
plants? Current Opinion in Plant Biology 14, 225–231.
Tudela, J.A., Marín, A., Martínez-Sánchez, A., Luna, M.C., and Gil, M.I. 2013.
Preharvest and postharvest factors related to off-odours of fresh-cut ice-
berg lettuce. Postharvest Biology and Technology 86, 463–471.
Turrell, F., and Klotz, L. 1940. Density of stomata and oil glands and incidence
of water spot in the rind of Washington navel orange. Botanical Gazette
101, 862–871.
Tyree, M.T., and Dixon, M.A. 1986. Water stress induced cavitation and
embolism in some woody plants. Physiologia Plantarum 66, 397–405.
Umezawa, T., Sugiyama, N., Mizoguchi, M., Hayashi, S., Myouga, F.,
Yamaguchi-Shinozaki, K., Ishihama, Y., Hirayama, T., and Shinozaki,
K. 2009. Type 2C protein phosphatases directly regulate abscisic acid-
activated protein kinases in Arabidopsis. Proceedings of the National
Academy of Sciences of the United States of America 106, 17588–17593.
Vahisalu, T., Kollist, H., Wang, Y.F., Nishimura, N., Chan, W.Y., Valerio, G.,
Lamminmäki, A., Brosché, M., Moldau, H., Desikan, R., Schroeder, J.I.,
and Kangasjarvi, J. 2008. SLAC1 is required for plant guard cell S-type
anion channel function in stomatal signaling. Nature 452, 487–491.
Vahisalu, T., Puzõrjova, I., Brosché, M., Valk, E., Lepiku, M., Moldau, H.,
Pechter, P., Wang, Y.S., Lindgren, O., Salojärvi, J., Loog, M., Kangasjarvi,
J., and Kollist, H. 2010. Ozone-triggered rapid stomatal response
involves the production of reactive oxygen species, and is controlled by
SLAC1 and OST1. Plant Journal 62, 442–453.
Vainio, H., and Bianchini, F. 2003. Fruit and Vegetables. IARC Handbooks of
Cancer Prevention. World Health Organization, Geneva.
Van Doorn, W.G. 1997. Water relations of cut flowers. Horticultural Reviews
18, 1–85.
Van Doorn, W.G. 2012. Water relations of cut flowers: an update. Horticultural
Reviews 40, 55–106.
van Meeteren, U. 1978. Water relations and keeping-quality of cut gerbera
flowers 2. Water balance of ageing flowers. Scientia Horticulturae 9,
189–197.
van Meeteren, U. 1989. Water relations and early leaf wilting of cut chrysan-
themums. Acta Horticulturae 261, 129–135.
van Meeteren, U., Rezaei Nejad, A., and Harbinson, J. 2009. Effect of (changes
in) air humidity on transpiration and (adaptation of) stomatal closure of
Tradescantia leaves during water stress. Acta Horticulturae 847, 115–122.
Veierskov, B., and Kirk, H.G. 1986. Senescence in oat leaf segments under
hypobaric conditions. Physiologia Plantarum 66, 283–287.
Velez-Ramirez, A.I., van Ieperen, W., Vreugdenhil, D., and Millenaar, F.F. 2011.
Plants under continuous light. Trends in Plant Science 16, 310–318.
Vitagliano, C., and Hoad, G.V. 1978. Leaf stomatal resistance, ethylene evolu-
tion and ABA levels as influenced by (2-chloroethyl) phosphonic acid.
Scientia Horticulturae 8, 101–106.

213
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Vlad, F., Rubio, S., Rodrigues, A., Sirichandra, C., Belin, C., Robert, N., Leung,
J., Rodriguez, P.L., Laurière, C., and Merlot, S. 2009. Protein phospha-
tases 2C regulate the activation of the Snf1-related kinase OST1 by
abscisic acid in Arabidopsis. Plant Cell 21, 3170–3184.
Waller, J.M., Lenné, J.M., and Waller, S.J. 2001. Plant Pathologist’s Pocketbook.
CABI Publishing, Wallingford, UK.
Wang, P., and Song, C.P. 2008. Guard-cell signaling for hydrogen peroxide
and abscisic acid. New Phytologist 178, 703–718.
Wang, Y., Fu, X.Z., Liu, J.H., and Hong, N. 2011a. Differential structure and
physiological response to canker challenge between ‘Meiwa’ kum-
quat and ‘Newhall’ navel orange with contrasting resistance. Scientia
Horticulturae 128, 115–123.
Wang, Y., Li, L., Ye, T., Zhao, S., Liu, Z., Feng, Y.Q., and Wu, Y. 2011b. Cytokinin
antagonizes ABA suppression to seed germination of Arabidopsis by
down regulating ABI5 expression. Plant Journal 68, 249–261.
Wardlaw, C. 1936. Studies in tropical fruits. II. Observations on internal gas
concentrations in fruit. Annals of Botany 50, 655–676.
Waterland, N.L., Campbell, C.A., Finer, J.J., and Jones, M.L. 2010. Abscisic
acid application enhances drought stress tolerance in bedding plants.
HortScience 45, 409–413.
Weaver, L.M., and Amasino, R.M. 2001. Senescence is induced in individually
darkened Arabidopsis leaves, but inhibited in whole darkened plants.
Plant Physiology 127, 876–886.
Wigger, J., Phillips, J., Peisker, M., Hartung, W., Zur Nieden, U., Artsaenko,
O., Fiedler, U., and Conrad, U. 2002. Prevention of stomatal closure by
immunomodulation of endogenous abscisic acid and its reversion by
abscisic acid treatment: Physiological behaviour and morphological fea-
tures of tobacco stomata. Planta 215, 413–423.
Wilkinson, S., and Davies, W.J. 2002. ABA-based chemical signaling: The co-
ordination of responses to stress in plants. Plant, Cell and Environment
25, 195–210.
Wilkinson, S., Clephan, A.L., and Davies, W.J. 2001. Rapid low temperature-
induced stomatal closure occurs in cold-tolerant Commelina communis
leaves but not in cold-sensitive tobacco leaves, via a mechanism that
involves apoplastic calcium but not abscisic acid. Plant Physiology 126,
1566–1578.
Williams, M., Brown, M.A., Vesk, M., and Brady, C. 1994. Effect of posthar-
vest heat treatments on fruit quality, surface structure, and fungal dis-
ease in Valencia oranges. Animal Production Science 34, 1183–1190.
Williamson, V.G., and Milburn, J.A. 1995. Cavitation events in cut stems kept
in water: Implications for cut flower senescence. Scientia Horticulturae
64, 219–232.
Willmer, C., and Johnston, W. 1976. Carbon dioxide assimilation in some aer-
ial plant organs and tissues. Planta 130, 33–37.

214
 Stom ata a n d P ostharvest P hy s i o l o g y

Wingler, A., Von Schaewen, A., Leegood, R.C., Lea, P.J., and Quick, W.P. 1998.
Regulation of leaf senescence by cytokinin, sugars, and light effects on
NADH-dependent hydroxypyruvate reductase. Plant Physiology 116,
329–335.
Wise, R., Olson, A., Schrader, S., and Sharkey, T. 2004. Electron transport is
the functional limitation of photosynthesis in field grown pima cotton
plants at high temperature. Plant, Cell and Environment 27, 717–724.
Xu, Z., and Zhou, G. 2008. Responses of leaf stomatal density to water sta-
tus and its relationship with photosynthesis in a grass. Journal of
Experimental Botany 59, 3317–3325.
Xu, Z.J., Nakajima, M., Suzuki, Y., and Yamaguchi, I. 2002. Cloning and char-
acterization of the abscisic acid-specific glucosyltransferase gene from
adzuki bean seedlings. Plant Physiology 129, 1285–1295.
Xue, H.W., Chen, X., and Mei, Y. 2009. Function and regulation of phospho-
lipid signaling in plants. Biochemical Journal 421, 145–156.
Yakushiji, H., Morinaga, K., and Nonami, H. 1998. Sugar accumulation and
partitioning in Satsuma mandarin tree tissues and fruit in response to
drought stress. Journal of the American Society for Horticultural Science
123, 719–726.
Yan, F., Sun, Y., Song, F., and Liu, F. 2012. Differential responses of stomatal
morphology to partial root-zone drying and deficit irrigation in potato
leaves under varied nitrogen rates. Scientia Horticulturae 145, 76–83.
Yoshida, R., Hobo, T., Ichimura, K., Mizoguchi, T., Takahashi, F., Aronso, J.,
Ecker, J.R., and Shinozaki, K. 2002. ABA-activated SnRK2 protein kinase
is required for dehydration stress signaling in Arabidopsis. Plant and
Cell Physiology 43, 1473–1483.
Yoshida, R., Umezawa, T., Mizoguchi, T., Takahashi, S., Takahashi, F., and
Shinozaki, K. 2006. The regulatory domain of SRK2E/OST1/SnRK2.6
interacts with ABI1 and integrates abscisic acid (ABA) and osmotic
stress signals controlling stomatal closure in Arabidopsis. Journal of
Biological Chemistry 281, 5310–5318.
Zeevaart, J.A.D. 1974. Levels of ({+/−}) abscisic acid and xanthoxin in spinach
under different environmental conditions. Plant Physiology 53, 644–648.
Zeiger, E. 2000. Sensory transduction of blue light in guard cells. Trends in
Plant Science 5, 183–185.
Zeng, W., Melotto, M., and He, S.Y. 2010. Plant stomata: a checkpoint of host
immunity and pathogen virulence. Current Opinion in Biotechnology 21,
599–603.
Zhang, K., and Gan, S.S. 2012. An abscisic acid-AtNAP transcription factor-
SAG113 protein phosphatase 2C regulatory chain for controlling dehy-
dration in senescing Arabidopsis leaves. Plant Physiology 158, 961–969.
Zhang, K., Xia, X., Zhang, Y., and Gan, S.S. 2012. An ABA-regulated and Golgi-
localized protein phosphatase controls water loss during leaf senes-
cence in Arabidopsis. Plant Journal 69, 667–678.

215
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Zhang, X., Takemiya, A., Kinoshita, T., and Shimazaki, K.I. 2007. Nitric oxide
inhibits blue light-specific stomatal opening via abscisic acid signaling
pathways in Vicia guard cells. Plant and Cell Physiology 48, 715–723.
Zhang, X., Wang, H., Takemiya, A., Song, C.P., Kinoshita, T., and Shimazaki,
K. 2004. Inhibition of blue light-dependent H+ pumping by abscisic acid
through hydrogen peroxide-induced dephosphorylation of the plasma
membrane H+ -ATPase in guard cell protoplasts. Plant Physiology 136,
4150–4158.
Zhao, X., Qiao, X.R., Yuan, J., Ma, X.F., and Zhang, X. 2012. Nitric oxide inhib-
its blue light-induced stomatal opening by regulating the K+ influx in
guard cells. Plant Science 184, 29–35.
Zhou, R., Cutler, A.J., Ambrose, S.J., Galka, M.M., Nelson, K.M., Squires,
T.M., Loewen, M.K., Jadhav, A.S., Ross, A.R., and Taylor, D.C. 2004. A
new abscisic acid catabolic pathway. Plant Physiology 134, 361–369.
Zhu, M., Dai, S., and Chen, S. 2012. The stomata frontline of plant interac-
tion with the environment—Perspectives from hormone regulation.
Frontiers in Biology 7, 96–112.
Zhu, S.Y., Yu, X.C., Wang, X.J., Zhao, R., Li, Y., Fan, R.C., Shang, Y., Du, S.Y.,
Wang, X.F., Wu, F.Q., Xu, Y.H., Zhang, X.Y., and Zhang, D.P. 2007. Two
calcium-dependent protein kinases, CPK4 and CPK11, regulate abscisic
acid signal transduction in Arabidopsis. Plant Cell 19, 3019–3036.
Zieslin, N. 1989. Postharvest control of vase life and senescence of rose flow-
ers. Acta Horticulturae 261, 257–264.
Ziv, M., Schwartz, A., and Fleminger, D. 1987. Malfunctioning stomata in
vitreous leaves of carnation (Dianthus caryophyllus) plants propagated
in vitro: Implications for hardening. Plant Science 52, 127–134.

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Chapter 7

Water Loss from

Harvested Horticultural
Commodities
Mikal E. Saltveit
University of California, Davis, California

Abstract 218
7.1 Importance of Water Loss 219
7.2 Properties of Water 220
7.3 Psychrometrics: Behavior of Water in Air 221
7.4 Transpiration: Diffusion of Water Vapor 224
7.5 Resistance to Diffusion of Water Vapor 226
7.6 Measurement of Transpiration 226
7.6.1 Weight Loss 227
7.6.2 Direct Measurement 227
7.6.3 Diffusion Porometer 227
7.7 Factors Affecting Water Loss 227
7.7.1 Commodity Factors 227
7.7.1.1 Surface-to-Volume Ratio 228
7.7.1.2 Routes of Water Loss 228
7.7.1.3 Anatomy of the Evaporating Surface 229
7.7.1.4 Physiological State of the Commodity 230

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7.7.1.5 Cultivar 230

7.7.1.6 Cultural Conditions 231
7.7.2 Environmental Factors 231
7.7.2.1 Humidity 231
7.7.2.2 Diffusion Shells and Air Velocity 231
7.7.2.3 Temperature 232
7.7.2.4 Atmospheric Pressure 232
7.8 Methods to Reduce Water Loss 232
7.8.1 Handling Techniques 232
7.8.2 Proper Refrigeration Design 232
7.8.3 Packaging 233
7.8.4 Waxing 234
7.8.5 Film Wraps 234
7.8.6 Curing 234
7.9 Conclusions and Future Perspectives 235
References 235

Abstract
Water loss from harvested horticultural commodities will continue to be
a problem as long as warm products need to be cooled and kept cold
during storage and marketing. Even a small amount of water loss can
adversely affect product quality, marketability, and storability. Apart
from the loss of fresh weight that accompanies water loss, a few percent
water loss can stimulate physiological changes that hasten senescence.
The physics of heat transfer in a mechanical refrigeration system neces-
sitates that the air in a cold room will have a vapor pressure less than
that of the commodity. This inherent difference in the vapor pressure
between water near the surface of fresh fruits, vegetables, and ornamen-
tals and the ambient cooling air is the prominent force that determines
the rate of water loss. Increasing the relative humidity of the surround-
ing air or the resistance of the surface of the commodity to the diffu-
sion of water vapor will decrease the rate of water loss. This can be done
by applying a wax or edible coating to the surface of whole or fresh-cut
commodities, by enclosing them in a package that raises the ambient
humidity, and by using harvest and postharvest practices that reduce
injuries (e.g., bruises, wounds, abrasions, etc.) that decrease the ability
of the surface to retard water loss. Preharvest factors (e.g., cultivar selec-
tion, growing environment, cultural practices, and maturity at harvest)
can have significant effects on rates of water loss. Familiarity with the

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psychrometric chart provides insights into the relationships among vapor

pressures, relative humidity, and product temperature. Rapid cooling of
warm-harvested commodities and maintaining a high relative humidity
in the cold room is essential to minimize the initial and subsequent level
of water loss.

7.1  Importance of Water Loss

Since most fresh fruits, vegetables, and ornamentals are 90%–95% water,
even a few percent loss of water can produce a significant change in their
postharvest behavior (Table 7.l). Water loss at levels much below those
that produce visible symptoms can influence many of the physical and
physiological changes occurring in harvested horticultural crops. These
effects range from physical (e.g., wilting, shriveling) and economic (e.g.,
weight loss) changes, to subtle visual and physiological (e.g., softening,
senescence) changes. While most effects are deleterious (e.g., reduction of
wound healing in roots and tubers), some may be beneficial (e.g., reduction
of chilling injury symptoms in some sensitive fruit).
Water loss can be a major source of economic loss during the post-
harvest period. The quality losses associated with these changes can result

Table 7.1  Effects of Water Loss on the Postharvest Behavior of Harvested

Horticultural Commodities
Water Loss (%) Possible Change in Postharvest Behavior
0.5 Greater activity of pectolytic enzymes (e.g.,
polygalacturonase).
1.0 Increased CO2 and C2H4 production; faster ripening,
yellowing, and abscission. Reduced wound healing
(periderm formation).
2.0 Reduced turgor, increased ABA content; reduced
susceptibility to chilling injury. Accelerated loss of
volatiles.
3.0 Reduced severity of certain physiological disorders such
as senescent breakdown. Loss of membrane integrity.
4.0 Faster loss of vitamins A and C, and flavor. Discoloration
of mechanical injuries.
5.0 Loss of color intensity and gloss. Accentuation of pitting
associated with chilling injury. Wilting and shriveling.
6.0 Loss of textural quality, e.g., softening, limpness,
flaccidity, and loss of crispness and juiciness.

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from a reduction in grade, quality, or price of the commodity. In addition,

the loss of water reduces fresh weight, which directly becomes a reduction
in value for products sold by weight. Water loss may also reduce the total
package weight so that the package may be rejected as no longer meet-
ing the grade standard. Repacking product to meet grade standards can be
very expensive.
An important function of water is to maintain the outward form of
perishable products. The cells in a commodity are like tiny balloons that
are osmotically inflated with water under pressure. The cell wall constrains
this turgor pressure, producing rigid, inflexible cells that contribute to the
texture and appearance of tissues. Water loss produces a reduction in tur-
gor pressure and the shrinkage of cells that becomes visually apparent as
wilting of leafy vegetables, flaccidity of other vegetables, and shriveling and
wrinkling of fruits.
Water loss can also be indirectly responsible for deterioration during
transport and marketing. Many perishables are “bulge” packed so that each
piece is tightly constrained within the box. The inevitable loss of water after
harvest reduces the size of the commodity, and if the individual commodi-
ties are not immobilized by packing material, they can be knocked about
within the package during handling and transport. This can lead to vari-
ous types of transit damage (e.g., scuffing of soft fruits, “roller” bruising of
pears, and shattering of grapes).

7.2  Properties of Water

Water is a remarkable chemical whose properties are fundamental to the
processes of life. The electrical potential surrounding a water molecule is
not uniformly distributed. The partial opposite charges on the oxygen and
hydrogen atoms in a water molecule (i.e., the asymmetric charge separa-
tion that produces polarity) makes water an effective solvent for a wide
range of chemicals, including many biologically important molecules.
Noncovalent hydrogen bonds between the hydrogen of one molecule and
the oxygen of an adjacent water molecule contribute to the exceptional
thermodynamic properties of water. Water has a high heat capacity (i.e.,
it takes a lot of energy to change water’s temperature: 1 kcal/kg/°C) and
an extremely high heat of vaporization (i.e., it takes a lot of energy to turn
liquid water into vapor: 540 kcal/kg/°C). These two properties help sta-
bilize the plant’s temperature (e.g., reducing the rate of freezing on cool
nights, and preventing overheating through evaporative cooling of tissue
exposed to direct sunlight). Water is also incompressible, so it can be an
important structural component of cells.
Water loss from perishable crops results primarily from evaporation
of liquid water from the surfaces of cells and diffusion of this water vapor

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 WATER LOSS

away from the tissue. The epidermis of all terrestrial plants is covered with
a waxy cuticle that evolved to significantly reduce diffusion of water vapor
from the tissue. However, this coating also reduces the diffusive exchange
of O2 and CO2 with the external environment. Specialized cells in the epi-
dermis of photosynthetically active leaves (i.e., guard cells) surround a
pore (i.e., the stoma) that breaches the epidermal barrier. Guard cells can
actively change their shape and size to alter the size of the stomatal open-
ing to balance the import of CO2 against the loss of water vapor. In other
organs, static ventilation systems such as lenticels allow diffusion of CO2
and O2 across the epidermis while minimizing water loss. The cuticle may
be modified during growth, development, and ripening to alter its water
diffusion characteristics. The rate of water loss, that is, transpiration, is
therefore controlled by active (i.e., stomata) and passive (e.g., lenticels)
routes of water loss. Transpiration in actively growing plants also provides
the driving force for water uptake by the roots and movement through the
xylem to the leaves.

7.3  Psychrometrics: Behavior of Water in Air

The behavior of water in the vapor phase is termed psychrometrics (i.e.,
properties of water vapor in air at different temperatures). When liquid
water is in contact with air, an equilibrium is established between mole-
cules that evaporate from the liquid’s surface and those that remain in the
liquid phase. At equilibrium, the amount of water vapor in the air above liq-
uid water is dictated by the temperature and the solute concentration. The
contribution of this latter factor is typically ignored since the solute con-
centration of water evaporating from the cell’s surface is usually very low.
In the vapor phase, water vapor acts just like any other gas. The total
pressure of the air in contact with liquid water is usually 1 atm (i.e., 101 kPa,
1013 mbar, 760 mmHg) and is the sum of the partial pressures of the com-
ponent gasses (i.e., N2, O2, Ar, CO2, H 2O, etc.).
The relationship between all the variables involved (see list below)
can be displayed in a psychrometric chart (Figure 7.1). Careful study of
the psychrometric chart reveals a great deal about the behavior of water
molecules in air, which is of some importance to postharvest physiologists
and technologists. Terms used in the psychrometric chart are listed and
defined below:

Dry bulb temperature: The horizontal axis of the chart is the

temperature of a dry thermometer in air.
Vapor pressure (VP): The vertical right-hand axis of the chart
shows the partial pressure of water vapor in the air (e.g., in kPa or
some other measure of pressure).

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Relative humidity (%)

100 80 60
4.5
30
4.0

3.5
25

)
°C
e(
3.0

ur
rat

Vapor pressure (kPa)

pe
tem 20 2.5
lb
bu
et
W

2.0
15

1.5
10

5 1.0
0
100
Relative 80 0.5
60
humidity 40
(%) 20
0 0.0
0 5 10 15 20 25 30 35 40 45
Dry bulb temperature (°C)

Figure 7.1  A psychrometric chart showing the interrelationships among dry

and wet bulb temperatures, vapor pressures, and relative humidity.

Wet bulb temperature: The left-to-right upward curving lines

show the temperature of a thermometer whose bulb is covered
with ventilated water-soaked gauze.
Dew point temperature: The horizontal lines extending from the
right axis and intersecting the wet bulb temperature line at the
dew point, the temperature of a surface upon which liquid water
will condense.
Saturation vapor pressure (SVP): The maximum amount of
water vapor (expressed as a partial pressure) that air at a specific
temperature can hold. It is found by extending a horizontal line
from the wet bulb temperature to the right boundary of the chart.
Relative humidity (RH): The ratio of the actual water content
of the air to the maximum possible water content under any par-
ticular temperature. The RH lines are represented by the equally
spaced curved lines that slope upward from left to right. The RH
can be expressed for a particular air sample and temperature as

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 WATER LOSS

RH  =  (VP/SVP) × 100, where RH = percent of maximum water

vapor in the air sample at that temperature, SVP = saturation vapor
pressure at 100% RH and that temperature, and VP = vapor pres-
sure in the air sample at that temperature.

Let’s use a few examples to see how we can use the psychrometric chart
(Figure 7.1).
The relative humidity (RH) can be determined by finding the inter-
section of the line extending upward from the dry bulb temperature with
the downward-sloping line from the wet bulb temperature. For example,
air with a dry bulb temperature of 25°C and a wet bulb temperature of 18°C
has a RH of 50% (point A on Figure 7.2). The vapor pressure (VP) of that
50% RH air can be determined by following the horizontal line to its inter-
section with the vertical axis at 1.6 kPa (point B on Figure 7.2). The dew
point temperature of 14°C can be determined by following the horizontal

Relative humidity (%)

100 80 60
4.5
30
4.0

3.5
)
°C

25
e(

3.0
ur
rat

Vapor pressure (kPa)

pe
tem

20 2.5
lb
bu

18
50
et

2.0
W

15 A B
C 14
1.6
1.5
10

5 1.0
0
100
Relative 80 0.5
60
humidity 40
(%) 20
0 0.0
0 5 10 15 20 25 30 35 40 45
Dry bulb temperature (°C)

Figure 7.2  Example of how the psychrometric chart can be used to find the
vapor pressure and dew point from measurements of the wet and dry bulb
temperatures (refer to text for detailed description).

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

line left to its intersection with the wet bulb temperature curve (point C on
Figure 7.2).
In another example, the RH of air with a dry bulb temperature of 15°C
and a wet bulb temperature of 12°C will be 70%, with a vapor pressure of
1.2 kPa and a dew point of 10°C.
If a sample of warm moist air is cooled (move left along a horizontal
line), its relative humidity increases until it reaches saturation (at the left-
most line, the wet bulb temperature curve). The cooled air is now at its dew
point. Further removal of heat will result in condensation of water from the
air onto the cool surface. In contrast, the RH of air rapidly decreases with
increasing temperature (move right along the horizontal line from the wet
bulb temperature curve). This means that at warm temperatures, air can
hold much more water vapor than at cool temperatures. The water holding
capacity of air just about doubles with every 11°C rise in temperature.
If a cold commodity is placed in a warm room, the air in contact with
the commodity will become cooler, and depending on the initial RH of the
air, water may condense on the surface of the fruit. In contrast, if a warm
commodity is placed in a cool atmosphere, the air in contact with the fruit
will become warmer (move to the right along the horizontal lines) and its
RH will decrease, thereby facilitating water evaporation from the tissue.

7.4  Transpiration: Diffusion of Water Vapor

For any gas, the rate at which it diffuses between two points is proportional
to the difference in partial pressures between the points and the diffusive
resistance of the intervening material. The rate at which water vapor dif-
fuses from a commodity is therefore proportional to the difference between
the vapor pressure at the surface of the commodity and the vapor pressure
of the surrounding air. The vapor pressure of water molecules in the air
can be determined from the psychrometric chart for any temperature and
RH. Because air inside a commodity is in equilibrium with the almost pure
water in the cell walls, the vapor pressure in the air spaces within tissue is
close to the saturated vapor pressure at that temperature. The vapor pres-
sure difference between the tissue and the outside air can therefore be cal-
culated with the equation
VPD = SVPinternal − VPexternal
where VPD = vapor pressure difference between the tissue and the
­surrounding air, SVPinternal = saturation vapor pressure of air at the tem-
perature of the tissue, and VPexternal = vapor pressure in the external air at
its temperature and RH.
The VPD can be used to determine the rates of water loss for different
combinations of commodity, air temperature, and RH using the following

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 WATER LOSS

equation (these calculations are only correct when other conditions, such
as barometric pressure, nature of the commodity’s surface, and air velocity
past the product remain constant).
J = K × VPD
where J = rate of water loss (g/h, % FW/day, etc.), K = ­proportionality
­constant (a function of many features of the commodity that will be
discussed later), and VPD = vapor pressure difference, or deficit (kPa,
mmHg, mbar, etc.).
For example, consider a harvested head of lettuce at 25°C that loses
1% of its fresh weight when held for 1 hour (i.e., J = 1%/h) in a room at 25°C
and 80% RH. What will be its initial and final rates of weight loss during
cooling to 4°C in a store at 80% RH (Figure 7.3)?
From the psychrometric chart we can determined that at 25°C, the
­saturation VP is 3.2 kPa (point A on Figure 7.3). The VP of the 25°C  air

Relative humidity (%)

100 80 60
4.5
30
4.0

3.5
)
(°C

25
A
re

3.0
tu
ra

Vapor pressure (kPa)

pe
tem

20 2.5
lb
bu
et

VPD = 3.2 – 0.6 = 2.6

W

2.0
15

1.5
10

5 1.0
0 B
100
Relative 80 0.5
humidity
60
40
VPD = 0.8 – 0.6 = 0.2
(%) 20
0 0.0
0 5 10 15 20 25 30 35 40 45
Dry bulb temperature (°C)

Figure 7.3  Example of how the psychrometric chart can be used to calcu-
late the vapor pressure deficit (VPD) experienced by a head of lettuce placed
into a cold room as it cools from 25°C to 4°C (refer to text for detailed
description).

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

at  80% RH is 80% of the saturation VP or about 2.5 kPa, and the VPD is
the difference of the two values, or about 0.7 kPa (i.e., 3.2 − 2.5 = 0.7).
Substituting in the equation J = K × VPD, we can calculate the proportional-
ity constant K [K = (1%/h)/(0.7 kPa) = 1.4% FW/kPa-h].
The VP of 100% RH air at 4°C is 0.8 kPa, while 80% RH air at the
same temperature has a VP of 0.6 kPa (0.8 × 0.80) (point B of Figure 7.3).
When the 25°C lettuce is initially put in the 80% RH 4°C cold room, the VPD
between the tissue and the air will be 2.6 kPa (i.e., 3.2 − 0.6). The initial rate
of water loss will therefore be 2.6 kPa × 1.4% FW/kPa-h = 3.6% FW/h. This
initial high rate of water loss is one reason why cooling to the correct stor-
age temperature should be done as quickly as possible. Once the product
is cooled to 4°C, the VP of the lettuce decreases to 0.8 kPa and the VPD
between the product and the air decreases to 0.2 kPa (0.8 − 0.6). The new
rate of water loss is 0.2 kPa × 1.4% FW/kPa-h = 0.28% FW/h. This amounts
to a 92% reduction in the rate of water loss.

7.5  Resistance to Diffusion of Water Vapor

The transpiration equation ( J = K × VPD) is a specific example of Fick’s law
of gas diffusion, which in the case of the diffusion of water vapor states that
J = (Area/Resistance) × (SVPinternal − VPexternal) = (A/R) × (VPD)
where J = rate of water vapor loss, A = surface area over which water vapor
is being lost to the external environment, R = coefficient of resistance to
water vapor diffusion, and VPD = vapor pressure difference, or deficit (kPa,
mmHg, mbar, etc.).
Once VPD is minimized by reducing the temperature and increasing
the RH around the commodity, the resistance to diffusion (R) is the most
important constant for determining susceptibility of different products to
the loss of water vapor. Because R is a resistance coefficient, higher val-
ues indicate lower rates of water loss. The composition of the cuticle, and
its resistance to diffusion, may change in response to growing conditions,
during development, and during postharvest handling. Waxing and film
coatings are some postharvest practices that are used to increase the com-
modities’ resistance to water vapor loss.

7.6  Measurement of Transpiration

Measurements of the rates of water loss are as important to the postharvest
physiologist as are measurements of respiration and ethylene production.
Several methods are commonly used; the most important are described in
the following subsections.

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 WATER LOSS

7.6.1  Weight Loss

Since almost all weight loss is due to water loss, the simplest method to deter-
mine the rate of water loss is to periodically weigh the commodity and cal-
culate the rate of water loss over time. In a few cases (e.g., long-term storage
of commodities such as onions), the dry weight loss due to respiration may
contribute to total weight loss and can be a source of errors with this method.

7.6.2  Direct Measurement

Measurement of changes in relative humidity (RH) or dew point of air in
a container enclosing a known weight of commodity can be used to deter-
mine the transpiration rate; this is similar to the method used in a static
or flowing system to measure the rate of respiration (see Chapter 5). This
method may be useful for researchers wishing to study water loss in com-
modities under controlled atmospheres or when CO2 and C2H4 production
is also being measured.

7.6.3  Diffusion Porometer

The diffusion porometer, designed to measure diffusive resistance of leaves
(and thereby to estimate opening of the stomata), can be used to measure
the diffusive resistance of those perishable commodities that lose water
fairly readily.

7.7  Factors Affecting Water Loss

As noted above, the equation used to describe water loss from perishables is,
in fact, a simplified version of an important law governing the diffusion of all
gases, Fick’s law of diffusion. This law states that diffusion of gases across a
surface is governed by the area of the surface, the resistance of the surface to
diffusion, and the concentrations of gases on either side of the surface. The
last term is governed by the temperature and RH in the storage environment,
while the other two terms are dependent on the properties of the commodity.

7.7.1  Commodity Factors

Commodities differ in size, shape, composition, physiology, and the compo-
sition and topology of the epidermis and cuticle. Many of these properties
can be significantly changed due to the conditions under which they were
grown and under which they are handled after harvest. The same cultivar

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can have markedly different rates of water loss depending on where and
when it was grown and how it was handled during and after harvest.

7.7.1.1  Surface-to-Volume Ratio

The rate at which water is lost from a commodity is a function of its sur-
face area. If one calculates water loss as a percentage of the initial product
weight for commodities whose surfaces have the same resistance to water
vapor diffusion, the rate of water loss will be a function of the surface
area-to-volume ratio of each commodity. Simple formulae for calculating
surface area and volume (Table 7.2) of commodities reveal major differ-
ences in the surface-to-volume ratio between large and small commodi-
ties (Table 7.3). If the rate of water loss per unit surface area is identical
for large and small units of a commodity, then the large item will still
lose less water per unit weight than the smaller item because it has less
surface area to volume than the smaller unit. For any commodity, smaller-
sized items will lose water at a faster rate than larger items.

7.7.1.2  Routes of Water Loss

There are two routes by which perishable commodities lose water.
Epidermal cells on the surface of the commodity can lose water directly
through their surface. The outer surfaces of all plant parts are covered with
water-repellant coverings: cuticle in aerial organs and suberin in roots and

Table 7.2  Equations for the Calculation of the Area and Volume for a Sphere
(Tomato), a Cylinder with Spherical Ends (Cucumber), and A right Circular
Cone with One Hemispherical End (Carrot).
Sphere Cylinder Cone
Area 4πr2 2πLr + 4πr 2 πr'r2 + L2 + 2πr2
Volume 4/3πr 3 πLr2 + 4/3πr3 1/3πLr2 + 2/3πr3

Table 7.3  Area and Volume of Some Commodities

Length Radius Area Volume
Commodity (cm) (cm) (cm2) (cm3 or mL) Area/Volume
Tomato (small) 4 201 268 0.75
Tomato (large) 8 804 2,144 0.38
Cucumber (small)  8 2 151 134 1.12
Cucumber (large) 16 4 603 1,072 0.56
Carrot (small)  8 2  89 50 1.78
Carrot (large) 16 4 358 402 0.89

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 WATER LOSS

tubers. Water losses through the epidermal cells occur by a relatively com-
plex process whereby water molecules must dissolve into the epidermal cov-
ering, diffuse through it, and then evaporate into the outside atmosphere.
The rate at which evaporation occurs is often limited by the solubility of
water in the outer surface coating.
The surface coating is very effective in retarding water loss. For exam-
ple, evaporation from the cells inside the leaf is 10-fold higher than from
cells on the leaf surface. In very heavily cutinized or suberized organs, very
little water escapes by direct evaporation from the surface cells. Instead,
most water is lost in the vapor phase through apertures in the surface of
the organ, such as stomata, lenticels, and wounds. The rate of evaporation
is affected not only by the route of evaporation, but also by the architecture
of the evaporating surface.

7.7.1.3  Anatomy of the Evaporating Surface

Important routes of evaporation and anatomical features affecting its
rate are

Stomata: Specialized organelles common to all green tissues of

higher plants; stomata are designed to facilitate inward diffusion
of CO2 during photosynthesis, but to minimize water loss at times
of low light or during water stress. Typically, the lower surfaces of
leaves are much more richly supplied with stomata than the upper
surface. Stomata will normally close when a tissue or organ is
under water stress, but can remain open during sudden reduction
in temperature (such as occurs during forced-air cooling), thus
aggravating water loss during storage.
Lenticels: In large organs, there is a danger that insufficient O2
will diffuse into the tissues to prevent anaerobiosis because of the
diffusive resistance of the cuticle. Lenticels are growths of unsu-
berized subepidermal cells that erupt through the epidermis and
facilitate the passive ventilation of bulky tissues. Changes in their
structure and thickness in response to environmental changes
during development and after harvest can modify their diffusive
properties.
Surface imperfections: In many commodities, especially after
harvest, water escapes through various imperfections in the epi-
dermis. Some of these imperfections may be the result of abnor-
mal growth; others, such as stem scars, open calyx, and blossom
scars, are normal anatomical features. Wounds of any sort also
result in increased water loss. Bruising, abrasion, and most
importantly, actual rupture of the epidermis result in greatly
accelerated water loss.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Cuticular waxes: The chemical composition and a­ rchitecture

of the cuticular surface can dramatically affect water loss.
Thickness of the cuticle obviously affects the rate of cuticular
diffusion of water. Waxes deposited on the surface of maturing
fruits and vegetables frequently have a complex architecture
composed of randomly oriented plates and fibers. These obstruc-
tions to airflow produce diffusion shells above the surface that
retard water loss.
Trichomes: Water loss in many plant parts is reduced by the pres-
ence of surface appendages called trichomes. These may be single
celled, multicelled, tufted, star shaped, or branched. Although
trichomes increase the surface area of an organ, they also trap
saturated water vapor around it and reduce air velocities past the
evaporating surface. Breakage of hairs or trichomes in posthar-
vest handling can aggravate water loss from commodities such as
peaches, kiwifruit, and zucchini.
Architecture: The effect of the surface-to-volume ratio on the
rate of water loss has been discussed already. In many plant
parts, the way in which the organ is constructed may also have
a dramatic effect on the rate of water loss. A striking example
is the difference between leaf lettuce and head cabbage. Both
are leafy vegetables, but the condensed cabbage offers little of
its leaf surface to evaporation. In compact leafy vegetables such
as cabbage or onion, the rate of water loss may be limited by the
ability of the vascular system to provide water to the outer evapo-
rating leaves. The desiccated scale leaves of onion or wrapper
leaves of cabbage can further reduce the rate of water loss from
the commodity.

7.7.1.4  Physiological State of the Commodity

It is obvious that young vegetative tissues, such as watercress, with plen-
tiful stomata and thin cuticles, lose a great deal more water than heavily
suberized storage organs such as potato. More subtle changes in physiolog-
ical state, such as ripening, can also strongly affect the rate of water loss.
The effect of ripening on the composition and structure of the surface wax
of apples has been alluded to before. A similar effect is probably responsible
for the difference in the water loss from dark green and yellow lemons.

7.7.1.5 Cultivar
The many structural and chemical differences between different cultivars
can substantially affect weight loss.

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 WATER LOSS

7.7.1.6  Cultural Conditions

Seasonal differences in environmental conditions during the growing
period may also affect the rate of weight loss from harvested commodities.
The porosity and diffusivity of the epidermis and cuticle can vary signifi-
cantly between the same commodity grown in hot versus cool and humid
versus dry environments. There are even differences in the rate of water
loss from fruit grown in the shaded inside of the tree and those grown on
the sunny periphery of the tree.

7.7.2  Environmental Factors

The postharvest environment can have a substantial effect on the rate of
water loss from harvested commodities. Environmental factors are particu-
larly important during long-term storage.

7.7.2.1 Humidity
The importance of the water vapor content of the air on water loss has
been emphasized already. Lower-humidity environments also frequently
adversely affect the quality of stored commodities. Because of the natural
cycling of temperatures in cold storage rooms, a compromise must be made
between the danger of condensation of water on cold fruit during the warm
part of the cycle in rooms with very high RH, and unacceptable levels of
water loss if the RH is maintained at levels safe for the entire cycle.

7.7.2.2  Diffusion Shells and Air Velocity

When water vapor diffuses beyond the epidermis, it has not yet entirely
escaped from the influence of the commodity. Thin surface layers of satu-
rated air, termed diffusion shells, which may be pronounced in commodi-
ties with hairs or elaborately sculptured wax deposits on their surface,
provide a significant further barrier to diffusion of water vapor. For this
reason, the rate at which the ambient air is moving past the commodity can
strongly affect the rate of water loss.
As the air velocity increases, the rate of water loss increases in a
hyperbolic fashion, eventually reaching a plateau when, presumably, the
diffusion shells have been completely swept away. In the design of cooling
and storage facilities, and in evaluating postharvest handling systems, it is
important to choose an appropriate compromise between air velocities that
will maintain adequate temperature control and those that will exacerbate
the loss of water from the product.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

7.7.2.3 Temperature
The close interdependence of temperature, VP, and RH is implicit in the
psychrometric chart, and the importance of temperature in the VPD-driven
loss of water from perishable commodities cannot be overemphasized.
Some studies have shown a substantial effect of temperature on water loss,
even allowing for the changes in VPD. Enhanced respiration and changes
in the composition of the cuticle at warmer temperatures may account for
these observations.

7.7.2.4  Atmospheric Pressure

Reducing the total atmospheric pressure (e.g., at high altitudes or under
hypobaric conditions during storage or transport) reduces the partial pres-
sure of water vapor and other gases in the air. This, in turn, accelerates
water loss (outward diffusion of water vapor) from fresh produce. This is
the basis of vacuum cooling, where a lowered pressure facilitates evapora-
tive cooling of the commodity.

7.8  Methods to Reduce Water Loss

A wide range of tools are available to the postharvest technologist for reduc-
ing the rate of water loss from perishable commodities. These techniques
are designed to reduce water loss either by reducing VPD or by increasing
the resistance of the surface-to-water diffusion.

7.8.1  Handling Techniques

The primary factors in reducing water loss are the care and rapidity with
which the commodity is removed from the plant, packed, and brought to
the optimum storage temperature. Rough handling will increase surface
damage and water loss through the ruptured epidermis. Tardiness in han-
dling, packing, and cooling will increase the time over which the product is
exposed to high VPDs and result in high water loss.

7.8.2  Proper Refrigeration Design

It is desirable to maintain high relative humidity in the cold room, especially
for the long-term storage of perishables. In general, the highest humidities
are obtained by jacketing the storeroom to reduce heat infiltration and loss
of humidity. There should be more than enough refrigeration capacity to

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 WATER LOSS

remove heat from all anticipated sources. Evaporators should be as large as

possible to permit the removal of the required amount of heat without having
to be run at a large temperature differential. The transfer of heat from the
storage room air to the cold evaporator coils is governed by the temperature
differential between them and the volume of air passing over the coils. If the
evaporator is too small, a large temperature differential will be needed to
transfer enough energy to maintain the storage temperature. A very low coil
temperature will be needed to produce the needed large temperature dif-
ferential. Water will condense on the cold coils, thereby lowering the water
content of the cold air exiting the evaporator. As the air warms to the storage
room temperature, its relative humidity will fall to undesirable levels.
In recent years, much has been claimed about the “humifresh” sys-
tems, in which the storage room air is circulated through baths or curtains
of water at or near 0°C. These systems allow very high humidity and very
uniform temperatures, but can be limited in their capacity and ability to
maintain temperatures at or near freezing.
Humidifiers raise the humidity by injecting mist or fog into the stor-
age room, but unless the refrigeration plant is designed to accommodate
the extra load of condensate, the evaporator coils may become covered
with ice, resulting in an actual reduction in humidity due to lowered coil
temperatures. Modern equipment can be fitted with hot gas bypass valves,
which allow the surplus capacity of the refrigeration system to bypass the
condenser when the commodities have attained store temperature and the
refrigeration load is lighter. This system can also divert warm gas to peri-
odically defrost the evaporator coils.
The importance of air velocity to weight loss was noted above. Storage
rooms should be designed to maintain proper product temperatures without
excessive rates of air movement. High air velocities are often used initially
to rapidly cool produce (e.g., forced-air precooling). Thereafter, convective
systems are usually adequate to maintain temperatures and will prevent
excessive weight loss.

7.8.3 Packaging
Placing commodities in packages of any sort obviously changes their geom-
etry with respect to weight loss. The larger the package, the smaller is its
surface area in relation to its volume. This is because the surface area
increases as a function of the dimensions of the package squared, while
the volume increases as a function of the dimensions cubed. In effect, the
package reduces the VPD within the package because the air inside the
package will be at high RH. To further reduce water loss, some products
are packed in polyethylene or other plastic liners, usually ventilated to avoid
anaerobic or high CO2 conditions in the package. Airflow through holes in

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

the package to maintain the desired product temperature and gas concen-
trations will nullify this benefit.
Certain containers, such as wooden or plain fiberboard boxes, can
absorb a substantial amount of water, thereby decreasing the initial VP in
the package. It is not feasible to equilibrate the containers with the storage
atmosphere, and waxed fiberboard cartons may be too expensive for many
commodities. Under these circumstances, waxed paper or polyethylene lin-
ers can substantially reduce weight loss for little extra cost.

7.8.4 Waxing
An alternative route to those noted above for reducing water loss is to raise
the resistance of the fruit surface to diffusion of water vapor by the applica-
tion of wax. Waxing is widely used as a treatment for fruits, frequently for
cosmetic reasons rather than to reduce weight loss. The waxes employed
are proprietary formulations and probably comprise largely waxes derived
from petrochemicals. Remember that although waxing can slightly reduce
the rate of water loss, temperature (even allowing for differences in the
VPD) can have a far greater effect than waxing. Waxing can also modify
the  internal atmosphere of perishable commodities. There is a danger
that the use of surface coatings can produce toxic internal levels of CO2,
­particularly when the commodity is returned to ambient temperatures after
transportation or storage.

7.8.5  Film Wraps

In recent years, there has been much interest in the use of individual film
wraps for packaging and marketing some fruits and vegetables. This tech-
nology has become a commercial proposition with grapefruit and lemons.
Shrink-wrapping individual grapefruit in high-density polyethylene film
reduced the rate of water loss from the fruits. This technique also reduces
the occurrence of decay and several disorders common in harvested citrus.
The relative contributions of reduction in water loss, isolation of individual
fruits, and modification of the fruit’s internal atmosphere to this beneficial
response have not yet been determined.

7.8.6 Curing
In some commodities, the formation of new or heavier deposits of suber-
ized cells on the surface will substantially increase their resistance to water
loss. Onions, potatoes, and sweet potatoes are frequently cured to improve

234
 WATER LOSS

their postharvest life and reduce water loss. High temperatures and RH
often assist in the suberization process that occurs during curing.

7.9  Conclusions and Future Perspectives

Reducing water loss is an important consideration in deciding which post-
harvest practices are used. Understanding how the physical laws of gas
diffusion govern water loss and how the concepts visualized in the psychro-
metric chart can be used to calculate the rate of water loss is essential to
formulating and implementing postharvest practices that will minimize
the detrimental effects of water loss. Future practices and technologies to
minimize water loss must be devised to act within the constraints imposed
by these physical laws and also within government regulations. Consumer
preferences will continue to identify those factors that must be either mini-
mized or maximized to optimize product quality. Economic considerations
are also important since no practice will be widely adopted unless it offers a
significant economic advantage over existing systems. The current desire
to consume more locally grown food may minimize the importance of
reducing the rate of water loss since the time from harvest to consumption
would be reduced. In contrast, the desire for more exotic horticultural com-
modities, the desire that seasonal crops be available year-round, and the
basic economics of large-scale production may all contribute to increase
transit times (and levels of water loss) as the distance between areas of pro-
duction and consumption increases. Future research should study package
designs that minimize water loss and cuticular and epidermal properties
that maximize the resistance to water vapor diffusion.

References
Kader, A.A. 2002. Postharvest Technology of Horticultural Crops. 3rd ed.
Division of Agriculture and Natural Resources, University of California,
Oakland.
Kader, A.A., and Saltveit, M.E. 2003. Respiration and gas exchange. In Bartz,
J.A., and Brecht, J.K. (eds.), Postharvest Physiology and Pathology of
Vegetables. Marcel Dekker, New York, pp. 7–30.
Kays, S.J., and Paull, R.E. (eds.). 2004. Postharvest Biology. Exon Press,
Athens, GA.
Wills, R.B.H., McGlasson, B., Graham, D., and Joyce, D. (eds.). 2007.
Postharvest: An Introduction to the Physiology and Handling of Fruit,
Vegetables and Ornamentals. CAB International, Wallingford, UK.
Woods, J.L. 1990. Moisture loss from fruits and vegetables. Postharvest News
and Information 1, 195–199.

235
Chapter 8

Lysophospholipids and
Postharvest Quality
of Fruits, Vegetables,
and Cut Flowers
Domingos P.F. Almeida
Universidade de Lisboa, Lisbon, Portugal

Abstract 238
8.1 Introduction 238
8.2 Lipids as Signal Molecules in Plant Senescence and
Stress Response 239
8.2.1 Chemistry 239
8.2.2 Metabolism 240
8.3 Modulation of Postharvest Quality by Lysophospholipids 241
8.3.1 Effects on Color 242
8.3.2 Effects on Texture 242
8.3.3 Other Effects on Postharvest Quality-Related
Features 246
8.4 LPE Treatment Condition 247
8.5 Improvement of Postharvest Quality by
Lysophospholipids: Potential and Limitations 248
8.6 Conclusions and Future Perspectives 249
References 249

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Abstract
Regulatory lipid molecules are involved in plant responses to ­environmental
stresses and developmental cues. Phosphatidic acid is one of the best-
known signaling lipids in plants, but other lipidic molecules are involved in
signal transduction pathways. Therefore, the potential exists for the tech-
nological or biotechnological use of signaling lipids as tools to modulate the
features of fresh produce with postharvest relevance. One attempt has been
made with the exogenous application of lysophosphatidylethanolamine
(LPE) to crops and produce. Modulation of postharvest quality by lyso-
phospholipids has been documented in dozens of horticultural commodi-
ties. Reported effects include delayed senescence and improved color and
texture, among other postharvest quality-related features. Despite the evi-
dence for the regulatory effects of lysophospholipids, two decades of trials
of LPE applications to horticultural produce have not yet been conclusive
regarding the development of lipids as plant growth regulators to improve
postharvest quality. Different approaches can be envisioned to harness the
potential for quality modulation by regulatory lipids: genetic engineering
to alter endogenous levels of regulatory lipids, pharmacological treatments
to target the biosynthesis of the signal transduction pathways of regulatory
lipids, and exogenous application of formulations containing signaling or
regulatory lipids. The improvement of our fundamental understanding of
lipid-mediated signaling and metabolic regulation is likely to result in novel
technological applications aimed at modulated produce quality.

8.1 Introduction
The importance of membranes on the postharvest quality of perishable
plant organs has long been recognized for their structural role, cellu-
lar compartmentation, and site of intense metabolic activity (Marangoni
et al., 1996). However, in addition to these functions, lipids have emerged
as an important class of regulatory and signaling molecules, and their role
in responses to environmental stresses and developmental cues is now
evident in plants and mammalians. Many regulatory lipids are related to
membrane phospholipids, as anabolic precursors or degradation products.
Phosphatidic acid is one of the best-known signaling lipids in plants (Wang
et al., 2006), but other lipidic molecules are involved in signal transduction
pathways. Therefore, the potential exists for the technological or biotech-
nological use of signaling lipids as tools to modulate the features of fresh
produce with postharvest relevance. One attempt has been made with the
exogenous application of lysophosphatidylethanolamine (LPE) to crops
and produce.

238
 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

This chapter provides an overview of the potential use of lysophos-

pholipids as tools to improve the quality attributes of fruits, vegetables, and
cut flowers; summarizes two decades of trials of LPE applications to horti-
cultural produce; and highlights the difficulties encountered in developing
lipids as plant growth regulators.

8.2  Lipids as Signal Molecules in Plant

Senescence and Stress Response

8.2.1 Chemistry
Phospholipids are structural components of cell membranes. In addition,
phospholipids and their catabolic products are important messengers in
the regulation of plant growth, development, and response to stress. These
molecules are chemically derived from a glycerol backbone esterified to
fatty acids in the sn-1 and sn-2 positions and phosphorylated at the sn-3 posi-
tion (Figure 8.1). The diversity of phospholipids is made possible by the
nature of the fatty acids and the head groups in the structure. The total
phospholipid composition of Arabidopsis leaves can be taken as represen-
tative of plant cells. The head groups most common in plants are choline,
ethanolamine, and glycerol (Miquel and Browse, 1992; Li et al., 2006), with
lower quantities of inositol and serine (Li et al., 2006). The fatty acids most

PLA1
PAT-PLA
H O

H C O C R1
O

R2 C O C H
O
PLA2

H C O P O X

H O

PLC PLD

Figure 8.1  Structure of a generic phospholipid with the acyl chains R1 and
R2 and the head group X, indicating the covalent bond hydrolyzed by PLA,
PLC, and PLD.

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

commonly found in plant phospholipids are 16:0, 16:3, 18:2, and 18:3, with
a lower proportion of 16:1, 16:2, 18:0, and 18:1 (Miquel and Browse, 1992).
The sn-2 carbon is usually esterified to an unsaturated acyl chain.

8.2.2 Metabolism
Plant membrane phospholipids are hydrolyzed by phospholipases, a large
and diverse group of enzymes belonging to three major classes: phospholi-
pases A, C, and D. Phospholipase A (PLA) hydrolyzes the acyl chain from
the glycerol backbone at the sn-1 or sn-2 positions or both, releasing free
fatty acids and lysophospholipids as reaction products. Three subtypes
of PLA are defined based on their positional specificity. PLA 1 and PLA 2
specifically hydrolyze the ester bond at the sn-1 and sn-2 positions, respec-
tively, whereas patatin-like PLA (PAT-PLA) catalyzes the hydrolysis at
both positions. Phospholipase C (PLC) cleaves the linkage between the
phosphate and the glycerol, releasing a phosphoryl head group and adiacyl
glycerol. Phospholipase D (PLD) hydrolyzes the terminal phosphodiester
bond between the head group and the phosphate group, releasing phospha-
tidic acid. Each of these classes of phospholipases is encoded by a large
gene family and contains several isoforms that can be grouped according
to their sequence similarities and substrate specificities (Qin and Wang,
2002; Meijer and Munnik, 2003; Li et al., 2007, 2009; Tasma et al., 2008;
Chen et al., 2011). The hydrolytic products of phospholipase activity have
a wide range of roles in plant metabolism, including responses to abiotic
stresses (drought, salinity, freezing, wounding, ultraviolet radiation), dis-
ease responses, hormone effects (absicic acid and auxin), jasmonic acid
biosynthesis, nutrition, including nitrogen assimilation and response to
phosphorus starvation, cell growth and elongation, shoot gravitropism,
onset of senescence, intracellular trafficking, stomatal function, and cyto-
skeletal organization (Wang et al., 2006; Xue et al., 2009; Chen et al., 2011;
Boss and Im, 2012; Dong et al., 2012).
Considering the broad range of physiological processes that can be
mediated by phospholipid-derived molecules, technological applications of
these molecules to modulate postharvest quality attributes in fruits, veg-
etables, and ornamentals can be foreseen. Arguably, the most immediate
technology is the direct application of exogenous lipidic molecules to crops
or harvested produce. This approach has been attempted with LPE.
LPE is the lysophospholipid resulting from the hydrolysis of phospha-
tidylethanolamine (PE) at the sn-2 position catalyzed by PLA 2. LPE is natu-
rally present in small concentrations in plant tissues (Li et al., 2006). The
chemical nature of LPE depends on the acyl chain present at the sn-1 posi-
tion of the glycerol backbone. LPE containing a 14:0 acyl chain is seldom

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 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

present in plant tissues (Ryu et al., 1997), but the 16:0, 18:0, and 18:1 acyl
chains exist in plants.

8.3  Modulation of Postharvest Quality

by Lysophospholipids
The biochemical, physiological, and horticultural effects of the applications
of LPE to horticultural commodities have been recently reviewed (Amaro
and Almeida, 2013). The molecule has documented effects on the in vitro
activity of PLD and a few other enzymes, on ethylene biosynthesis, and on
lipid catabolism.
Of special interest for quality-related applications is the observation
that LPE inhibits the activity of PLD (presumably PLDα) in vitro (Ryu
et  al., 1997). The inhibition is concentration dependent and depends also
on the nature of LPE. The inhibitory effect is much higher with 18:1-LPE
than with 14:0-, 16:0-, or 18:0-LPE (Ryu et al., 1997). Despite this evidence,
PLD inhibition by exogenously applied LPE has not yet been demonstrated
in vivo in horticultural commodities, and the relevance of in vitro inhibition
of PLD by LPE to the physiological effects is not clear (Hong et al., 2009b).
If materialized in vivo, an inhibitory effect of LPE on PLD would provide
a means to regulate the production of phosphatidic acid and delay senes-
cence (Hong et al., 2009a).
LPE treatments also affect ethylene biosynthesis, but the reported
effects are inconsistent (Amaro and Almeida, 2013). Applied to ripe cran-
berry, a nonclimacteric fruit, 18:1-LPE significantly reduces the ethylene
production rate, but 14:0-LPE has no effect, while 16:0- and 18:0-LPE reduce
ethylene biosynthesis only slightly (Ryu et al., 1997). LPE is also reported to
reduce the ethylene production rate in flowers of carnation (Kaur and Palta,
1996) and snapdragon (Kaur and Palta, 1997), and in tomato fruit (Farag
and Palta, 1993a; Hong, 2006). However, LPE has also been reported to
increase the ethylene production rate in the climacteric fruit apple (Farag
and Palta, 1991c), banana (Ahmed and Palta, 2011a), and tomato (Hong
et al., 2001), and in the nonclimacteric cranberry (Farag and Palta, 1991c)
and red pepper (Kang et al., 2003). The effect of LPE on the ethylene pro-
duction rate may depend on the maturity stage, at least in the climacteric
tomato fruit (Altwies et al., 2002; Hong et al., 2002; Hong, 2006). The mech-
anism by which LPE interferes with ethylene biosynthesis is unknown, but
candidate sites of action are the expression of ACC synthase genes (Hong
et al., 2008), ACC oxidase activity (Hong et al., 2002; Hong, 2006), and the
ethylene signal transduction pathway (Cowan, 2009).
Lysophospholipid can act as molecular chaperones in vitro facilitating
the maintenance or recovery of protein tertiary structure (Kern et al., 2001).

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

In addition, several enzyme activities, other than PLD, seem to be affected

by exogenous treatments with LPE formulations. Transient increases in
acid invertase, phenylalanine ammonia lyase, endo β-1,3(4)-glucanase, and
peroxidase, and reduction in the activity of polyphenoloxidase, 3-hydroxy-
3-methylglutaryl coenzyme A reductase, and polygalacturonase have been
reported (Farag and Palta, 1992a; Mangat and Palta, 1995; Cowan et al.,
2006; Hong et al., 2007, 2009b). The effects of LPE on these enzymes are
likely exerted via interference with signal transduction or gene expression
(Alvarez-Venegas et al., 2006a, 2006b), rather than direct action upon the
enzymes (Hong et al., 2009a).
Considering the effects of LPE, mainly on PLD activity and ethylene
biosynthesis, the exogenous application of the substance should affect key
attributes of fruits, vegetables, and ornamentals. Table 8.1 summarizes the
reported effects of LPE treatments on plants and plant organs.

8.3.1  Effects on Color

Treatments with LPE interfere with the pigments and color of leaves and
fruits. Higher chlorophyll retention was observed in tomato leaves treated
with LPE (Farag and Palta, 1991b), and potato plantlets grown in medium
containing LPE had twice the chlorophyll content as ethylene-treated con-
trols (Özgen et al., 2005).
More abundant are the reports on fruit color. The preharvest spray
with LPE increases anthocyanin content and improves the color uniformity
of cranberry (Farag and Palta, 1991a; Özgen and Palta, 1999; Özgen et al.,
2004) and bicolored apples (Farag and Palta, 1991a). Tomato and pepper
crops, with climacteric and nonclimacteric fruit, respectively, sprayed with
LPE had faster fruit ripening and a higher proportion of red fruit at the time
of harvest (Farag and Palta, 1991b; Kang et al., 2003). In fresh-cut canta-
loupe fruit, however, a postprocessing treatment with LPE did not affect the
flesh color determined by carotenoid pigments (Amaro et al., 2012).

8.3.2  Effects on Texture

Improved firmness retention during storage as a consequence of LPE treat-
ments has been reported for cranberry, apple (Farag and Palta, 1991a,
1991c), and tomato fruit (Farag and Palta, 1993a). Also, excised banana peel
incubated with LPE at 100 mg L −1 remained firmer and with better tissue
integrity than tissue treated at lower LPE concentrations (Workmaster and
Palta, 1996). In contrast, the firmness of fresh-cut melon cubes vacuum-
infiltrated with 200 mg L −1 LPE was not affected by the treatment (Amaro
et al., 2012).

242
 Table 8.1  Summary of LPE Effects on Horticultural Commodities
Unaffected or
Species Increased Decreased or Delayed Inconsistent References
Fruits
Apple Firmness, color Respiration rate Farag and Palta (1991c)
uniformity, peel
anthocyanin content,
ethylene production
Banana Shelf life, fruit diameter, Senescence, respiration Fresh weight loss, Workmaster and Palta
marketable fruits, rate, soluble solids ethylene production (1996), Ahmed and
firmness content, electrolyte Palta (2010, 2011a)
leakage
Cranberry Fruit set, anthocyanin Chlorothalonil toxicity, Özgen and Palta (1999,
content, color ethylene production 2003, 2004), Farag and
uniformity, firmness Palta (1991c), Ryu et al.
L Y SO P H OS P H OLI P I D S

(1997)
Cantaloupe Aldehyde production Volatile alcohols and Amaro et al. (2012)
(fresh cut) esters
AN D

Grape Anthocyanin Hong et al. (2007)

concentration, soluble
solids content, firmness,
berry size
Loquat Fruit set, earliness of Pollen germination and Demirkeser et al. (2009)
flowering and maturity, pollen tube growth rates
pollen viability

(Continued )
P OST H AR V EST

243
244
Table 8.1  Summary of LPE Effects on Horticultural Commodities
Unaffected or
Species Increased Decreased or Delayed Inconsistent References
Pepper Ripening, ethylene Ethephon-induced injury, Kang et al. (2003), Hong
production chilling injury and Chung (2006)
P OST H AR V EST

Tomato Ripening, firmness, Ethylene production and Farag and Palta (1991b,
ethylene production respiration rates and 1991d, 1993a, 1993b),
(mature-green fruit), ACC oxidase activity in Mangat and Palta
respiration rate (mature ripening fruit, electrolyte (1995), Özgen et al.
green), ACC oxidase leakage, decay, (2000), Hong et al.
activity (mature green) polygalacturonase (2001, 2002), Altwies
activity, injury by et al. (2002), Hong
RI P ENING

ethephon, PLD activity (2006)

Flowers
Rose Vase life Flower opening Snider et al. (2003a)
Snapdragon Bud opening Weight loss, ethylene Kaur and Palta (1997)
production, ion leakage
Impatiens Number of open flowers, Snider et al. (2003b)
recovery from water
stress cycles
P H Y SIOLOG Y

Leaves and
cotyledons
Arabidopsis Freezing tolerance Rajashekar et al. (2006)
thaliana
 Philodendron PAL or insoluble acid Hong et al. (2009a)
cordatum invertase, ABA-induced
increase in
malonaldehyde,
chlorophyll
Potato Chlorophyll content Özgen et al. (2005)
(in vitro)
Radish Acid invertase, Polyphenoloxidase and Soluble acid invertase Cowan et al. (2006),
phenylalanine ammonia 3-hydroxy-3- activity, glucose and Hong et al. (2009b)
lyase, endo β-1,3(4)- methylglutaryl sucrose levels
glucanase and coenzyme A reductase
peroxidase (transient), activity
PAL and insoluble acid
invertase activity
L Y SO P H OS P H OLI P I D S
AN D
P OST H AR V EST

245
P OST H AR V EST RI P ENING P H Y SIOLOG Y

8.3.3  Other Effects on Postharvest Quality-Related Features

Other quality-related effects of LPE have been reported. The delayed senes-
cence of leaves and their protection against injury have been observed. LPE
delays the senescence of tomato leaves, lowering their rates of respiration
and ethylene production, maintaining fresh weight, and lowering electro-
lyte leakage in relation to untreated controls (Farag and Palta, 1991b). LPE
also protected the foliage of tomato (Özgen et al., 2000) and pepper (Kang
et al., 2003) plants against injury by ethephon, and increased the freezing
tolerance of Arabidopsis thaliana leaves (Rajashekar et al., 2006).
Floral crops and cut flowers also benefit from LPE treatments. LPE
enhanced bud opening, delayed weight loss, lowered ethylene production,
and reduced electrolyte leakage from the cut flowers of snapdragon d ­ uring
vase life in deionized water (Kaur and Palta, 1997). The same benefits of
LPE were reported in carnation flowers if treated until the developmen-
tal stage IV, but more developed flowers were not affected by LPE (Kaur
and Palta, 1996). In cut roses, a vase solution composed of LPE, the bio-
cide 8-hydroxyquinoline citrate, and sucrose as a respiratory substrate is
reported to increase the vase life by 30% in relation to a commercial vase
solution, despite the higher incidence of bent neck (Snider et al., 2003a). In
impatiens bedding plants subjected to cycles of water stress, LPE sprays
increased the number of open flowers and enhanced the crop’s ability to
recover from stress (Snider et al., 2003b).
In fruit crops and harvested fruit, LPE has been reported to improve
fruit setting, increase fruit size, decrease respiration rate and electrolyte leak-
age, and increase soluble solids content. Overall, the observations indicate
improved postharvest storability of LPE-treated fruit. Table grapes, sprayed
with 10 mg L −1 LPE 4 or 6 weeks after fruit set, yielded larger and firmer ber-
ries with higher soluble solids content than untreated controls (Hong et al.,
2007). In loquat trees, LPE spraying before bloom also improved fruit set
and anticipated the harvest date (Demirkeser et al., 2009). Higher fruit set
was also observed in LPE-treated cranberry (Özgen and Palta, 2003). In red
pepper, LPE applications increased the total number of harvested fruit by
30% and reduced the proportion of fruit with disease symptoms (Kang et al.,
2003). LPE also reduces pitting and decay during cold storage of green pep-
pers (Hong and Chung, 2006). Preharvest LPE spray mitigates the injury
caused by the fungicide chlorothalonil in cranberry (Özgen and Palta, 2003).
LPE improves storability of tomato fruit, reducing its respiration
rate and ethylene production during postharvest storage (Farag and
Palta, 1991b) and lowering electrolyte leakage (Farag and Palta, 1993a).
Improved membrane integrity with a reduction in electrolyte leakage and
ethylene biosynthesis was also observed in banana peel incubated in LPE
(Workmaster and Palta, 1996; Ahmed and Palta, 2011a).

246
 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

Inconsistencies exist in the reported effects of LPE on ethylene

­ roduction, respiration rate, and protection from decay on different com-
p
modities. LPE applied to apple and cranberry fruit by vacuum infiltration
or dipping stimulated ethylene production and had no effect on respiration
rate (Farag and Palta, 1991c), but in another study, lower ethylene produc-
tion and respiration rate were reported for cranberry fruit dipped in LPE
(Özgen and Palta, 1999). LPE spray reduces peel pitting but significantly
increases the decay of ‘Navel’ oranges compared with untreated fruit
(Alfarez et al., 2008).
Effects of LPE on fruit flavor have been insufficiently addressed,
but are anticipated based on the reported effects on ethylene and PLD.
LPE did not affect the total amount of volatile esters and alcohols in
fresh-cut melon cubes, but reduced the concentration of aldehydes
(Amaro et al., 2012).

8.4  LPE Treatment Condition

LPE has been exogenously applied in more than 30 trials to about 15 hor-
ticultural species, including whole plants, fruit, leaves, and cut flowers
(Amaro and Almeida, 2013). The application methods include preharvest
crop sprays, postharvest applications by dipping, and pulsing and vase solu-
tions for cut flowers. Experimental methods unsuitable for large-scale com-
mercial applications have also included vacuum infiltration and feeding of
fruit and leaves via the peduncle or petiole, respectively. Addition to the
in  vitro culture medium has also been used (Özgen et al., 2005; Ahmed
and Palta, 2011b). The concentrations of LPE used in exogenous applica-
tions range from 10 to 400 mg L −1, with most trials using 100 to 200 mg L −1.
These values are one to two orders of magnitude above the water solubility
of LPE (Wishart et al., 2009).
The described benefits of LPE (Table 8.1) include delayed leaf senes-
cence (Farag and Palta, 1993a; Özgen et al., 2005; Hong et al., 2009a),
stimulation of ripening in nonclimacteric table grape (Hong et al., 2007),
acceleration in color development and extension of shelf life in cranberry
(Özgen et al., 2004) and tomato (Farag and Palta, 1993b), and increased
vase life of cut flowers (Kaur and Palta, 1996, 1997; Snider et al., 2003a).
Unfortunately, in many trials, LPE was combined with other substances,
such as ethanol or surfactants, to promote the penetration of the solution
into the plant organs, which makes it difficult to ascribe the treatment
effects in vivo to LPE alone (Amaro and Almeida, 2013). In addition, the
nature of the LPE molecule used in the horticultural treatments, namely,
the acyl chain on the sn-1 position, is not always specified. The commer-
cially developed LPE, whose effects are discussed herein, is derived from

247
P OST H AR V EST RI P ENING P H Y SIOLOG Y

egg, which is the 18:0-LPE (Cowan, 2009). However, the 18:1-LPE has stron-
ger biochemical effects on ethylene and phospholipase D (Ryu et al., 1997).

8.5  Improvement of Postharvest Quality by

Lysophospholipids: Potential and Limitations
The mounting evidence for the signaling roles of phospholipid-derived
metabolites certainly opens the door for the development of technologi-
cal applications aimed at modulating crop responses and product qual-
ity, based on regulatory lipids. Different approaches can be envisioned:
(1) genetic engineering to alter endogenous levels of regulatory lipids,
(2) pharmacological treatments to target the biosynthesis of the signal
transduction pathways of regulatory lipids, and (3) exogenous applica-
tion of formulations containing signaling or regulatory lipids. As of yet,
only the latter approach has reached technological application in horti-
cultural crops and products, although without commercial success. Of
the three generic approaches to technology development, the exogenous
application of natural lipids to crops or products, as plant growth regula-
tors, is the most direct considering food safety and environmental reg-
ulations and the public perception of technologies applied to crops and
foods. It may not, however, be the most effective means of harnessing
the putative benefits of lipid-mediated quality modulation. The formula-
tions used so far contained one single phospholipid as the putative active
ingredient: LPE.
It is not clear whether the exogenous application of these phospho-
lipid metabolites is an effective means of inducing the putative effects on
a commercial scale. Moreover, the attribution of the reported effects to
the applied lysophospholipid cannot be unequivocally ascribed (Amaro
and Almeida, 2013). On the one side, lysophospholipids are amphipa-
thic molecules. Substances with amphipathic properties applied exog-
enously to plant organs elicit wound responses, affect biochemical
pathways (Dawson and Hemington, 1967), and interact with cellular
membranes (Hong et al., 2009b). Neutral lipids, phospholipids, stripped
plant oils, or even commercial plant oils have reported beneficial effects
in fruit ( Ju et al., 2000; Ju and Curry, 2000, 2001) that cannot be clearly
ascribed to one molecular component. On the other hand, it is not easy
to stabilize lipid molecules in an aqueous formulation for pre- or post-
harvest applications. Among the preparation methods described in the
literature, the addition of ethanol and detergents is reported to improve
the performance of LPE for postharvest dip applications (Özgen and
Palta, 1999). The differences in LPE preparation modes and the use
of excipients and adjuvants in the formulations may account for some

248
 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

of the reported effects attributed to LPE. Ethanol, per se, can delay
ripening-related changes and reduce decay in fruits (Margosan et al.,
1997; Chervin et al., 2005; Pesis, 2005).

8.6  Conclusions and Future Perspectives

The improvement of our fundamental understanding of lipid-mediated sig-
naling and metabolic regulation is likely to result in novel technological
applications aimed at modulated produce quality. Despite the in vitro inhibi-
tion of PLD and interference with ethylene production, the in vivo targets
of exogenous LPE remain unclear. The reported horticultural effects of
treatments with exogenous LPE are not always consistent with the putative
mode of action of LPE (Amaro and Almeida, 2013). The extent to which the
horticultural effects attributed to LPE are specific or generic responses to
amphipathic molecules or responses to excipients or additives in the formu-
lations is also unclear. Despite the inconsistencies in the published record
regarding the horticultural effects of LPE, other signaling or regulatory lip-
ids are likely to be developed into postharvest technologies to modulate the
quality of fruits, vegetables, and ornamentals. Whether other bioactive lip-
ids can be developed into plant growth regulation formulations remains an
open question. The pharmacological and genetic engineering approaches
to altering lipid-mediated metabolic regulation are currently farther from
practical applications, but the potential exists.

References
Ahmed, Z.F.R., and Palta, J.P. 2010. Lysophosphatidylethanolamine, a natu-
ral phospholipid, may retard senescence and improve the shelf life of
banana fruit. HortScience 45, 66. (Abstr.).
Ahmed, Z.F.R., and Palta, J.P. 2011a. A natural lipid, lysophosphatidyletha-
nolamine, may promote ripening while reducing senescence in banana
fruit. HortScience 46, 273.
Ahmed, Z.F.R., and Palta, J.P. 2011b. Hormone-like effect of a natural lipid,
lysophosphatidylethanolamine, can mitigate calcium deficiency injury
in potato shoot cultures. HortScience 46, 196.
Alfarez, F., Lluch, Y., and Burns, J.K. 2008. Phospholipase A2 and postharvest
peel pitting in citrus fruit. Postharvest Biology and Technology 49, 69–76.
Altwies, J., Guelzow, M., Hong, J.H., Rademacher, M., and Towse, J. 2002.
Potential use of lysophosphatidylethanolamine, a natural lipid, as a post-
harvest dip to accelerate ripening of mature green tomatoes packed for
fresh-market. In XXVI International Horticultural Congress, Toronto,
Canada, p. 250 [abstract].

249
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Alvarez-Venegas, R., Sadder, M., Hlavacka, A., Baluska, F., Xia, Y., Lu, G.,
Firsov, A., Sarath, G., Moriyama, H., Dubrovsky, J.G., and Avramova,
Z. 2006a. The Arabidopsis homolog of trithorax, ATX1, binds phospha-
tidylinositol 5-phosphate, and the two regulate a common set of tar-
get genes. Proceedings of the National Academy of Sciences USA 103,
6049–6050.
Alvarez-Venegas, R., Xia, Y., Lu, G., and Avramova, Z. 2006b. Phosphoinositide
5-phosphate and phosphoinositide 4-phosphate trigger distinct spe-
cific responses of Arabidopsis genes. Plant Signaling and Behavior 1,
140–151.
Amaro, A.L., and Almeida, D.P.F. 2013. Lysophosphatidylethanolamine
effects on horticultural commodities: A review. Postharvest Biology and
Technology 78, 92–102.
Amaro, A.L., Fundo, J.F., Beaulieu, J.C., Stein, R.E., Fernández-Trujillo, J.P.,
and Almeida, D.P.F. 2012. Lysophosphatidylethanolamine effects upon
volatiles and quality of fresh-cut cantaloupe melon. Acta Horticulturae
934, 959–965.
Boss, W.F., and Im, Y.J. 2012. Phosphoinositide signaling. Annual Review of
Plant Biology 63, 409–429.
Chen, G., Snyder, C.L., Greer, M.S., and Weselake, R.J. 2011. Biology and
biochemistry of plant phospholipases. Critical Reviews in Plant Science
30, 239–258.
Chervin, C., Westercamp, P., and Monteils, G. 2005. Ethanol vapours limit
Botrytis development over postharvest life of table grapes. Postharvest
Biology and Technology 36, 319–322.
Cowan, A.K. 2009. Plant growth promotion by 18:0-lyso-phosphatidyletha-
nolamine involves senescence delay. Plant Signaling and Behavior 4,
324–327.
Cowan, A.K., Leung, C., and Santori, C.H. 2006. Lyso-phosphatidyletha-
nolamine (LPE) as a plant bioregulator. Acta Horticulturae 727, 527–536.
Dawson, R.M.C., and Hemington, N. 1967. Some properties of purified
phospholipase D and specially the effect of amphipathic substances.
Biochemical Journal 102, 76–86.
Demirkeser, T.H., Polat, A.A., and Caliskan, O. 2009. Effect of natural lipid on
flowering, pollination traits and fruit set on loquat (Eriobotrya japonica
Lindl.). Asian Journal of Chemistry 21, 113–120.
Dong, W., Lv, H., Xia, G., and Wang, M. 2012. Does diacylglycerol serve as a
signaling molecule in plants? Plant Signaling and Behavior 7, 472–475.
Farag, K.M., and Palta, J.P. 1991a. Use of lysophosphatidylethanolamine, a
natural lipid, as an aid for fruit ripening and improving keeping quality.
In Proceedings of the 17th Annual Meeting of the Plant Growth Regulator
Society of America, Saint Paul, MS, USA pp. 135–137.
Farag, K.M., and Palta, J.P. 1991b. Use of lysophosphatidylethanolamine,
a natural lipid, to delay tomato fruit and leaf senescence. HortScience
26, 67 (Abstr.).

250
 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

Farag, K.M., and Palta, J.P. 1991c. Enhancing ripening and keeping quality of
apple and cranberry fruits using lysophosphatidylethanolamine, a natu-
ral lipid. HortScience 26, 67.
Farag, K.M., and Palta, J.P. 1991d. Improving postharvest keeping quality of
vine-ripened tomato fruits with a natural lipid. HortScience 26, 778.
Farag, K.M., and Palta, J.P. 1992. Evidence for a specific inhibition of the activ-
ity of polygalacturonase by lysophosphatidylethanolamine in tomato
fruit tissue: Implication for enhancing storage stability and reducing
abscission of the fruit. Plant Physiology 99, 54.
Farag, K.M., and Palta, J.P. 1993a. Use of lysophosphatidylethanolamine,
a natural lipid, to retard tomato leaf and fruit senescence. Physiologia
Plantarum 87, 515–524.
Farag, K.M., and Palta, J.P. 1993b. Use of natural lipids to accelerate ripen-
ing and enhance storage life of tomato fruit with and without ethephon.
HortTechnology 3, 62–65.
Hong, J.H. 2006. Lysophosphatidylethanolamine enhances ripening and pro-
longs shelf life in tomato fruit: Contrasting effect on mature green vs
red tomatoes. Horticulture, Environment, and Biotechnology 47, 55–58.
Hong, J.H., and Chung, G. 2006. Effects of application of lysophosphatidyleth-
anolamine on marketable yield and storability of red pepper (Capsicum
annuum L.). Horticulture, Environment, and Biotechnology 47, 243–246.
Hong, J.H., Altwies, J.R., Guelzow, M., and Palta, J.P. 2002. The influence
of lysophosphatidylethanolamine, a natural lipid, ethylene production
and ACC oxidase activity on mature green vs. red tomatoes. In XXVI
International Horticultural Congress, Toronto, Canada, p. 262 [abstract].
Hong, J.H., Arya, L.M., Altwies, J.R., and Palta, J.P. 2001. Stimulation of ripen-
ing of fresh-market tomato fruit by a postharvest dip in lysophosphati-
dylethanolamine (LPE), a natural lipid. HortScience 36, 598.
Hong, J.H., Chung, G., and Cowan, A.K. 2009a. Delayed leaf senescence by
exogenous lyso-phosphatidylethanolamine: Towards a mechanism of
action. Plant Physiology and Biochemistry 47, 526–534.
Hong, J.H., Chung, G., and Cowan, A.K. 2009b. Lysophosphatidylethano-
lamineenhanced phenylalanine ammonia-lyase and insoluble acid inver-
tase in isolated radish cotyledons. Plant Growth Regulation 57, 69–78.
Hong, J.H., Hwang, S.K., and Chung, G.H. 2008. Influence of lysophospha-
tidylethanolamine on reactive oxygen species, ethylene biosynthesis,
and auxin action in plant tissues. Korean Journal of Horticultural Science
and Technology 26, 209–214.
Hong, J.H., Hwang, H.K., Chung, G.H., and Cowan, A.K. 2007. Influence of
lysophosphatidylethanolamine application on fruit quality of Thompson
seedless grapes. Journal of Applied Horticulture 9, 112–114.
Ju, Z., and Curry, E.A. 2000. Stripped corn oil emulsion alters ripening,
reduces superficial scald, and reduces core flush in ‘Granny Smith’
apples and decay in ‘d’Anjou’ pears. Postharvest Biology and Technology
20, 185–193.

251
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Ju, Z., and Curry, E.A. 2001. Plant oil emulsions prevent senescent scald and
core breakdown and reduce fungal decay in ‘Bartlett’ pears. Journal of
the American Society for Horticultural Science 126, 358–363.
Ju, Z., Duan, Y., and Ju, Z. 2000. Mono-, di-, and tri-acylglycerols and phos-
pholipids from plant oils inhibit scald development in ‘Delicious’ apples.
Postharvest Biology and Technology 19, 1–7.
Kang, C.K., Yang, Y.L., Chung, G.H., and Palta, J.P. 2003. Ripening promotion
and ethylene evolution in red pepper (Capsicum annuum) as influenced
by newly developed formulations of a natural lipid, lysophosphatidyl-
ethanolamine. Acta Horticulturae 628, 317–322.
Kaur, N., and Palta, J.P. 1996. Prolonging the vase life of snapdragons and car-
nations with a natural lipid, lysophosphatidylethanolamine. HortScience
31, 636.
Kaur, N., and Palta, J.P. 1997. Postharvest dip in a natural lipid, lysophos-
phatidylethanolamine, may prolong vase life of snapdragon flowers.
HortScience 32, 888–890.
Kern, R., Joseleau-Petit, D., Chattopadhyay, M.K., and Richarme, G. 2001.
Chaperone-like properties of lysophospholipids. Biochemical and
Biophysical Research Communications 289, 1268–1274.
Li, G., Lin, F., and Xue, H.W. 2007. Genome-wide analysis of the phospholi-
pase D family in Oryza sativa and functional characterization of PLDβ1
in seed germination. Cell Research 17, 881–894.
Li, M., Hong, Y., and Wang, X. 2009. Phospholipase D- and phosphatidic
acid-mediated signaling in plants. Biochimica et Biophysica Acta 1791,
927–935.
Li, M., Welti, R., and Wang, X. 2006. Quantitative profiling of Arabidopsis polar
glycerolipids in response to phosphorus starvation: Roles of phospho-
lipases D ζ1 and D ζ2 in phosphatidylcholine hydrolysis and digalac-
tosyldiacylglycerol accumulation in phosphorus-starved plants. Plant
Physiology 142, 750–761.
Mangat, N.K., an Palta, J.P. 1995. Inhibition of polygalacturonase in tomato
pericarp tissue by lysophosphatidylethanolamine: Implications in fruit
shelf-life. HortScience 30, 889. (Abstr.)
Marangoni, A.G., Palma, T., and Stanley, D.W. 1996. Membrane effects
in posthar vest physiology. Postharvest Biology and Technology 7,
193–217.
Margosan, D.A., Smilanick, J.L., Simmons, G.F., and Delmer, J.H. 1997.
Combination of hot water and ethanol to control postharvest decay of
peaches and nectarines. Plant Disease 81, 1405–1409.
Meijer, H.J., and Munnik, T. 2003. Phospholipid-based signaling in plants.
Annual Review of Plant Biology 54, 265–306.
Miquel, M., and Browse, J. 1992. Arabidopsis mutants deficient in polyun-
saturated fatty acid synthesis: biochemical and genetic characterization
of a plant oleoyl-phosphatidylcholine desaturase. Journal of Biological
Chemistry 267, 1502–1509.

252
 L Y SO P H OS P H OLI P I D S AN D P OST H AR V EST

Özgen, M., and Palta, J.P. 1999. Using natural lipids to accelerate ripening
(uniform color development) and promote shelf life of cranberries. In
Wisconsin State Growers Association Summer Growers Meeting and Field
Day, pp. 33–34.
Özgen, M., and Palta, J.P. 2003. A natural lipid, lysophosphatidylethanolamine
(LPE), can mitigate adverse effect of fungicide, chlorothalonil, on fruit
set and yield in cranberries. Acta Horticulturae 628, 747–752.
Özgen, M., Farag, K.M., Özgen, S., and Palta, J.P. 2004. Lysophosphatidyle
thanolamine accelerates color development and promotes shelf-life of
cranberries. HortScience 40, 127–130.
Özgen, M., Palta, J.P., and Ryu, S.B. 2000. Mitigation of ethephon injury to
tomato plants by a natural lipid, lysophosphatidylethanolamine (LPE):
influence on the activity of phospholipase D (PLD). HortScience 35, 503.
Özgen, M., Park, S., and Palta, J.P. 2005. Mitigation of ethylene-promoted
leaf senescence by a natural lipid, lysophosphatidylethanolamine.
HortScience 40, 1166–1167.
Pesis, E. 2005. The role of the anaerobic metabolites, acetaldehyde and etha-
nol, in fruit ripening, enhancement of fruit quality and fruit deteriora-
tion. Postharvest Biology and Technology 37, 1–19.
Qin, C., and Wang, X. 2002. The Arabidopsis phospholipase D family:
Characterization of a calcium-independent and phosphatidylcholine-
selective PLDζ1 with distinct regulatory domains. Plant Physiology 128,
1057–1068.
Rajashekar, C.B., Zhou, H., Zhang, Y., Li, W., and Wang, X. 2006. Suppression
of phospholipase Dα1 induces freezing tolerance in Arabidopsis:
Response of cold responsive genes and osmolyte accumulation. Journal
of Plant Physiology 163, 916–926.
Ryu, S.B., Karlsson, B.H., Özgen, M., and Palta, J.P. 1997. Inhibition of phos-
pholipase D by lysophosphatidylethanolamine, a lipid-derived senes-
cence retardant. Proceedings of the National Academy of Sciences USA
94, 12717–12721.
Snider, A., Palta, J.P., and Peoples, T. 2003a. The potential use of lysophosphati-
dylethanolamine (LPE), a natural lipid, to lengthen the vase life of short-
lived ‘Lavande’ and ‘Sensation’ roses. Acta Horticulturae 624, 419–426.
Snider, A., Palta, J.P., and Peoples, T. 2003b. Use of lysophosphatidyletha-
nolamine (LPE), a natural lipid, to enhance opening and retention of
flowers on bedding plants experiencing water stress during retail sales.
Issues and advances in postharvest horticulture. Acta Horticulturae
628, 849–853.
Tasma, I.M., Brendel, V., Whitham, S.A., and Bhattacharyya, M.K. 2008.
Expression and evolution of the phosphoinositide-specific phospho-
lipase C gene family in Arabidopsis thaliana. Plant Physiology and
Biochemistry 46, 627–637.
Wang, X., Devaiah, S.P., Zhang, W., and Welti, R. 2006. Signaling functions of
phosphatidic acid. Progress in Lipid Research 45, 250–278.

253
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Wishart, D.S., Knox, C., Guo, A.C., Eisner, R., Young, N., Gautam, B., Hau,
D.D., Psychogios, N., Dong, E., Bouatra, S., Mandal, R., Sinelnikov, I.,
Xia, J., Jia, L., Cruz, J.A., Lim, E., Sobsey, C.A., Shrivastava, S., Huang,
P., Liu, P., Fang, L., Peng, J., Fradette, R., Cheng, D., Tzur, D., Clements,
M., Lewis, A., De Souza, A., Zuniga, A., Dawe, M., Xiong, Y., Clive, D.,
Greiner, R., Nazyrova, A., Shaykhutdinov, R., Li, L., Vogel, H.J., and
Forsythe, I. 2009. HMDB: A knowledge base for the human metabo-
lone. Nucleic Acid Research 37, D603–610.
Workmaster, B.A.A., and Palta, J.P. 1996. Using banana peel for bioassay
development to evaluate the retardation of senescence by lysophospha-
tidylethanolamine, a natural lipid. HortScience 31, 604.
Xue, H.W., Chen, X., and Mei, Y. 2009. Function and regulation of phospho-
lipid signalling in plants. Biochemical Journal 421, 145–156.

254
Chapter 9

Fruit Skin Color and

the Role of Pigments
during Fruit Ripening
Emrul Kayesh,1 Lingfei Shangguan,2
and M. Mofazal Hossain3
1Bangabandhu Sheikh Mujibur Rahman Agricultural
University, Gazipur, Bangladesh
2 Nanjing Agricultural University, Nanjing, China
3Bangabandhu Sheikh Mujibur Rahman Agricultural
University, Gazipur, Bangladesh

Abstract 256
9.1 Introduction 256
9.2 Economics of Color Fruit 258
9.3 Carotenoid Pigments in Fruits 259
9.4 Anthocyanin Pigments in Fruits 260
9.5 Grape as a Model System for Pigmentation Studies
in Fruit Crops 270
9.6 Case Studies 278
9.6.1 Apple 278
9.6.2 Strawberry 279

255
P OST H AR V EST RI P ENING P H Y SIOLOG Y

9.6.3 Tomato 280

9.6.4 Litchi 281
9.6.5 Chinese Bayberry 282
9.6.6 ‘Hass’ Avocados 283
9.6.7 Pear 283
9.6.8 Cherry 284
9.6.9 Kiwifruit 284
9.7 Pigments from Fruit to Human Health 284
9.8 Transgenic Plants Developed for Fruit Color 285
9.9 Conclusions and Future Perspectives 286
References 288

Abstract
The color of fruit is an important phase in its life cycle and results from the
presence of carotenoid and anthocyanin pigments. During coloration, sev-
eral physiological and biochemical changes take place through differential
expression of various genes that are developmentally regulated. Expression
and suppression of these genes contribute to various changes in the fruit
that make it visually attractive and edible. Pigments and surface topography
selectively absorb and refract incident visible light to produce a reflectance
spectrum characteristic of a particular fruit skin. The share of colored fruit
in the total fruit production is quite significant, which is contributing sev-
eral billion dollars annually. contributing several billion dollars annually.
This has encouraged scientists to study the pigmentation mechanism in
fruits. Pigmentation evokes several responses during coloration through
a signaling cascade, and thousands of genes are involved, with each gene
being a member of a multigene family. The transcripts are abundant in the
skins of cultivars with red skin, but rarely in skins of cultivars with other
fruit colors, suggesting that these genes have major roles in the determina-
tion of fruit skin color. In this chapter, various developments that have taken
place in the last decade with respect to identifying and altering the function
of ripening-related genes have been described. Taking clues from the stud-
ies in grape as a model fruit, a few case studies are reviewed.

9.1 Introduction
Fruits are an important part of the human supplementary diet, with the
quality of fruit being determined by a wide range of desirable character-
istics, such as food value, flavor, shelf life, and fruit color. Carotenoid and
anthocyanin are the most important pigments, being responsible for a

256
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

wide range of colors displayed by different plant organs, such as leaves,

fruits, and seeds. Accumulation of pigments in fruit skin is an important
determinant of fruit quality, with fruit skin color being one of the most
important qualities used as the basis for selection in breeding programs.
Nowadays, fruit skin color has become greatly diversified, and it is deter-
mined by the quality and composition of pigments. Carotenoids are some
of the most vital colored phytochemicals, occurring as all-trans- and
cis-isomers and accounting for the brilliant colors of a variety of fruits
and vegetables. Carotenoids extensively studied in this regard include
β-carotene, lycopene, lutein, and zeaxanthin. Coloration of fruits depends
on their growth maturity, concentration of carotenoid isomers, and food
processing methods. Carotenoids contribute to the yellow color found in
many fruits (Bartley and Scolnik, 1995; Lancaster et al., 1997). The col-
ors of fruits depend on conjugated double bonds and the various func-
tional groups contained in the carotenoid molecule (Rodriguez-Amaya
and Kimura, 2004). A study also reported that the greater the number
of conjugated double bonds, the higher the absorption maxima (λmax)
(Rodriguez-Amaya, 2001). As a result, the color ranges from yellow to
red to orange in many fruits and vegetables (Bartley and Scolnik, 1995;
Hornero-Méndez and Mínguez-Mosquera, 2000). Besides, esterification
of carotenoids with fatty acids can also occur during fruit ripening, which
may affect the color intensity (Minguez-Mosquera and Hornero-Mendez,
1994). Anthocyanins are the largest and most diverse group of plant pig-
ments derived from the phenylpropanoid pathway, ranging in color from
red to violet and blue (Tunen and Mol, 1991). They are water-soluble phe-
nolic compounds and part of a large and widespread group of plant flavo-
noids. There are less than 20 anthocyanidins (aglycones or chromophores
of anthocyanins), differing in the number and position of their hydroxyl
groups and methyl groups. Anthocyanidins are modified by glycosyl and
aromatic or aliphatic acyl moieties, resulting in hundreds of anthocyanin
molecules that differ in hue and stability. These pigments accumulate in
the vacuoles, and their stability and hue depend on intravacuolar condi-
tions such as pH, copigmentation with coexisting colorless flavonoids, and
formation of complexes with metal ions (Mazza and Miniati, 1993; Tanaka
and Ohmiya, 2008). Anthocyanins are located mainly in the epidermal tis-
sue, but are also present in the palisade and spongy mesophyll in leaves
and in the flesh of fruits and underground storage organs such as sweet
potato (Hughes et al., 2007; Sugawara and Igarashi, 2008).
Color development of fruit has been an important evolutionary
trait in plant fitness. Fruit color is a blend of carotenoids and flavonoids.
Flavonoids can act as UV and pathogen protectants as well as signal
­molecules in the interaction between bacteria and plants. Anthocyanin, a
subclass of flavonoids, has been implicated as a major color pigment in
the plants. Besides anthocyanin, other pigments, such as flavonols and

257
P OST H AR V EST RI P ENING P H Y SIOLOG Y

carotenoids, metal complexes, vacuolar pH, and cell shape, have been
known to influence the final pigmentation phenotype. The genetics and
biochemistry of the a­ nthocyanin biosynthetic pathway, a major branch
of flavonoid metabolism, have been well characterized in maize, petunia,
snapdragon, and more recently, Arabidopsis (Holton and Cornish, 1995;
Winkel-Shirley, 2001; Kim et al., 2003; Martens et al., 2010). The regula-
tion of anthocyanin biosynthesis has also been studied thoroughly and is
comprised of basic helix–loop–helix (bHLH) transcription factors, inter-
acting with R2R3 MYB transcription factors to activate either all or part of
the anthocyanin genes (Allan et al., 2008). Many studies have generated
transgenic plants with either increased concentration or altered composi-
tion of anthocyanin in flowers, or fruit by manipulating the expression of
the anthocyanin r­ egulatory genes (Quattrocchio et al., 1998; Davies et al.,
2003; Ben Zvi et al., 2008; Butelli et al., 2008; Tanaka and Ohmiya, 2008;
Oren-Shamir, 2009).
Fruit color is one of the characteristics that are important for both the
commercial and the aesthetic value of fruit. The mechanism of color forma-
tion in the fruit has not been studied extensively at the molecular level.
Color variation has a genetic basis that is species specific, with different
genes in the pathway having been found to be responsible for color varia-
tion in different species (Arakawa et al., 1999; Awad et al., 2000; Jaakola
et al., 2002; Ben-Yehudah et al., 2005). In this chapter, we review various
developments that have taken place in the last decade with respect to iden-
tification and alteration of functions of genes involved in fruit pigmentation
during fruit ripening.

9.2  Economics of Color Fruit

The share of colored fruit in the world’s total fruit production is very
­significant, which is contributing several billion dollars annually. Data
based on Food and Agriculture Organization (FAO) statistics (www
.faostat.fao.org) for the year 2012 on major colored fruit have been com-
piled.  Fruit produced by the top 20 countries is often referred to as the
total world production of that fruit, and therefore the figures quoted herein
follow this assumption. The total world production of the 22 major colored
fruit selected for this review in 2012 is 473,679,743 MT, of which tomato
occupies the top position, with 29.9% (141,400,629 MT), followed by banana
20% (95,595,965 MT), apple 15% (71,736,938 MT), grapes 14% (66,935,199
MT), and mangoes and mangosteens 7.40% (35,035,641 MT). On the
extreme end, cranberries and gooseberries have the least acreage har-
vested (0.14%) and least percentage of total production (0.12%) (Figure 9.1).
In terms of market value calculation, the total world fleshy fruit generates
more than 1.2 × 1012 USD, of which grapes top the list with 2.7 × 1010 USD,

258
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Avocados and cherries

1% 3% 2%
2%
4%
Persimmons, apricots, and
strawberries
30% 8%
Papayas
14% Plums and sloes

21% 15% Peaches and nectarines

Mangoes, mangosteens,
and guavas
Grapes

Figure 9.1  Percent share of major color fruit out of the total world p
­ roduction
in metric tons.

followed by tomato with 2.5 × 1010. While considering export potential,

apple, banana, tomato, and grapes fetch the maximum value compared to
other fruits (Kayesh et al., 2013).

9.3  Carotenoid Pigments in Fruits

Carotenoids are widely distributed in the cellular tissues of plants (Furr and
Clark, 1997). The distribution of carotenoids in human tissues is originated
from plant sources. Therefore, fruits constitute the major source of carot-
enoids in the human diet (Scott et al., 1996; Krinsky and Johnson, 2005). In
plants, carotenoids are found as fat-soluble and colored pigments (Simpson,
1985; Minguez-Mosquera et al., 1990). Carotenoids can be isolated from the
grana of chloroplasts in the form of carotenoprotein complexes, which give
various colors to the outer surfaces of the plants (Takyi, 2001). The visible
colors of the plant are due to the conjugated double bonds of carotenoids
that absorb light. The greater the number of double bonds, the more is the
absorbance of the red color wavelength. The occurrences of carotenoids in
plants are not as a single compound. Most of the carotenoids are bound with
chlorophyll, and a combination of carotene–chlorophyll and ­xanthophyll–
chlorophyll occurs often. The binding of carotenoids to chlorophylls can
give rise to a variety of colors in fruits; however, as fruit matures, the chloro-
phyll content decreases and results in colored carotenoid pigments (Marín
et al., 2004). Besides, a study has been carried out to improve carotenoids’
color retention during ripening (Markus et al., 1999). In nature, fruits have
less xanthophyll content than vegetables. Some fruits, such as papaya
and persimmon, have a high amount of xanthophylls (lutein and zeaxan-
thin). In fruits, β-carotene is found to be bound to either chlorophylls or

259
P OST H AR V EST RI P ENING P H Y SIOLOG Y

xanthophylls, forming chlorophyll–carotenoid complexes, which absorb

light in the orange or red light spectrum and give rise to green, purple, or
blue coloration (Wieruszewski, 2002). These complexes could decrease the
bioavailability of β-carotene and further weaken its bioefficacy for the con-
version to vitamin A. This setback, however, can be resolved by saponifying
the plant extract to yield all-trans-β-carotene in a free-state form (Yahia
et  al., 2006). Comprehensive data for the typical carotenoid contents in
fruits are given and summarized in Table 9.1. The data from this compila-
tion are useful for comparison of the ongoing study with other, previous
reports. Naturally occurring β-carotene, with 11 double bonds, is orange in
color (Rao and Rao, 2007). Takyi (2001) reported β-carotene occurs as an
orange pigment, while α-carotene is a yellow pigment, which can be found
in fruits. Yellow fruits that contain low or trace amounts of β-carotene are
mainly from the genera Ananas, Averrhoea, Citrus, Durio, Malus, Musa,
Nephelium, Pyrus, Rubus, and Vitis (Table 9.1).

9.4  Anthocyanin Pigments in Fruits

Anthocyanins are present in different plant organs, such as fruits, fl
­ owers,
stems, leaves, and roots (Brouillard, 1982). These pigments are normally
found dissolved uniformly in the vacuolar solution of epidermal cells.
However, in certain species, the anthocyanins are localized in discrete
regions of the cell vacuole, called anthocyanoplasts (Pecket and Small,
1980). The main sources of anthocyanins, as indicated in Table 9.2, are red
fruits, mainly berries and red grapes (Escribano-Bailón et al., 2004). The
content of anthocyanin may vary from fruit to fruit of the same type due
to different external and internal factors, such as genetic and agronomic
factors, intensity and type of light, temperature, processing, and storage.
For example, anthocyanin concentrations in red grapes are distinctively
variable, being able to reach values of up to 250 mg/100 g. The most com-
mon anthocyanidins in higher plants are delphinidin, cyanidin, petunidin,
pelargonidin, peonidin, and malvidin. The glycosides of the three nonmeth-
ylated anthocyanidins (delphinidin, cyanidin, and pelargonidin) are the
most abundant in nature, which represent 80%, 69%, and 50% of leaf, fruit,
and flower pigments, respectively. The distribution of the six most common
anthocyanidins in the edible parts of plants is cyanidin (50%), pelargonidin
(12%), peonidin (12%), delphinidin (12%), petunidin (7%), and malvidin (7%).
The most widespread anthocyanin in most fruits is cyaniding 3-glucoside
(Kong et al., 2003).
Color development of flower and fruit has been an important evolution-
ary trait in plant fitness. Anthocyanins, in combination with carotenoids and
chlorophylls, are responsible for almost all fruit coloration. Anthocyanins
belong to the diverse group of ubiquitous secondary metabolites known

260
 Table 9.1  Carotenoid Contents (mg/100 g fresh weight) of Some Common Fruits
Family Genus Species Name α-Carotene β-Carotene Lycopene References
ROLE

Anacardiaceae Mangifera indica L. Mango — 0.553 0.353 Setiawan et al.

(2001)
O F

— 1.71 (0.95) — Bhaskarachary

et al. (1995)
0.017 0.445 — Holden et al.
[0.001] [0.016] (1999)
var. ND 0.615 ND Tee and Lim
Black-gold (1991)
var. Gedong 0.061 3.267 — Hulshof et al.
P IGMENTS

(0.086) (2.075) (1997)

var. ND 0.19 (0.123) — Hulshof et al.
Manalagi (1997)
var. 0.067 1.606 — Hulshof et al.
Indramayn (0.005) (0.166) (1997)
D URING

var. Harum 0.055 1.08 (0.264) — Hulshof et al.

manis (0.001) (1997)
var. Golek 0.055 1.237 — Hulshof et al.
(0.003) (0.626) (1997)
F RUIT

(Continued )
RI P ENING

261
262
Table 9.1  Carotenoid Contents (mg/100 g fresh weight) of Some Common Fruits
Family Genus Species Name α-Carotene β-Carotene Lycopene References
Actinidiaceae Spondias dulcis L. Hog plum — 0.201 0.364 Hulshof et al.
Actinidia (1997)
deliciosa Kiwifruit ND 0.074 ND Yano et al.
P OST H AR V EST

C.F. Liang & [0.021] (2005)

A.R.
Ferguson
var.
Hayward
var. Zespri ND 0.092 ND Yano et al.
gold [0.008] (2005)
RI P ENING

Bromeliaceae Ananas comosus (L.) Pineapple ND 0.056 ND Yano et al.

Merr. [0.005] (2005)
ND 0.17 ND Lako et al.
(2007)
Caricaceae Carica papaya L. Papaya ND 0.23–1.981 1.477– Tee and Lim
5.75 (1991),
Setiawan et al.
(2001),
Bramley (2000)
P H Y SIOLOG Y

var. Fruit — 1.05 (0.44) — Yano et al.

tower ND 0.276 — (2005)
[0.245]
ND 0.409 2.481
[0.027] [0.692]
 var. Sun rise ND 1.981 1.477 Lako et al.
[0.059] [0.302] (2007)
var. Yellow ND 1.048 1.987
ROLE

sweet var. [0.026] [0.851]

Hawaiian ND 0.5 1.7
O F

Cucurbitaceae Citrullus lanatus Watermelon 0–0.76 0.14– 0.071– Yano et al.

Ebenaceae Diospyros (Thunb.) Persimmon 6.806 11.389 (2005)
Ericaceae Vaccinium Matsum & Blueberries ND 0.59 [0.033] 6.184 Lako et al.
Malvaceae Durio Nakai Durian [0.152] (2007)
sp. spp. — 0.253 —
zibethinus L. ND 0.129 0.415
[0.003] [0.013]
P IGMENTS

ND 0.035 ND
ND 0.027 ND
[0.005]
0.006 0.023 —
Moraceae Artocarpus heterophyllus Jackfruit ND 0.026–0.36 0.037 Charoensiri
Lam. et al. (2009)
D URING

— 0.16 (0.06) — Bhaskarachary

et al. (1995)
Musaceae Musa spp. Banana 0.005 0.021 — Yano et al.
[0.005] [0.014] (2005)
F RUIT

0.058 0.058 ND
[0.007] [0.006]

(Continued )
RI P ENING

263
264
Table 9.1  Carotenoid Contents (mg/100 g fresh weight) of Some Common Fruits
Family Genus Species Name α-Carotene β-Carotene Lycopene References
paradisiaca Banana — 0.097 0.114 Setiawan et al.
L. (2001)
var. Ambon
P OST H AR V EST

sapientum — 0.04 — Tee and Lim

Linn. (1991)
var. Emas
var. Tanduk — 0.092 — Tee and Lim
(1991)
Myrtaceae Psidium guajava L. Guava — 0.001 0.114 Setiawan et al.
(2001)
RI P ENING

Oxalidaceae Averrhoa var. Pink Pink guava — 0.001 — Bhaskarachary

Passifloraceae Passiflora carambola L. Starfruit (0.0001) et al. (1995)
edulis Sims Passion fruit — — 5.4 Bramley (2000)
ND 0.359 2.307 Bhaskarachary
[0.015] [0.058] et al. (1995),
5.027 4.383 Setiawan et al.
[0.08] [0.371] (2001), Yano
ND 0.028–0.042 0–0.042 et al. (2005),
ND ND ND Lako et al.
P H Y SIOLOG Y

0.035 0.53 — (2007),

ND 0.156 [0.02] 0.057 Charoensiri
[0.003] et al. (2009)
 Rosaceae Eriobotrya japonica Loquat — 0.207 — Granado-
(Thunb.) Lorencio et al.
Lindl. (2007)
ROLE

Fragaria ananassa Strawberry 0.005 — — Holden et al.

Duchesne (1999)
O F

Malus domestica Apple 0.001– 0.031– 0.209 Levy et al.

Borkh. 0.03 0.072 (1995),
var. Fuji ND 0.036 ND Dias et al.
[0.003] (2009)
Rosaceae Prunus armeniaca L. Apricot ND 2.554 0.005 Holden et al.
(1999)
salicina Lindl. Nectarine
P IGMENTS

var. Red Jim — 0.073 — Gil et al. (2002)

(0.016)
var. August — 0.128 — Gil et al. (2002)
red (0.005)
var. Spring — 0.085 — Gil et al. (2002)
D URING

bright (0.006)
var. May glo — 0.058 — Gil et al. (2002)
(0.005)
var. — 0.131 — Gil et al. (2002)
F RUIT

September (0.023)
red

(Continued )
RI P ENING

265
266
Table 9.1  Carotenoid Contents (mg/100 g fresh weight) of Some Common Fruits
Family Genus Species Name α-Carotene β-Carotene Lycopene References
persica (L.) Peach 0.001 0.097 — Holden et al.
Batsch [0.001] [0.013] (1999)
var. Summer — 0.04 (0.01) — Gil et al. (2002)
P OST H AR V EST

sweet
var. Snow — 0.008 — Gil et al. (2002)
king (0.002)
var. Santa — 0.049 — Gil et al. (2002)
Rosa (0.012)
var. — 0.057 — Gil et al. (2002)
Angeleno (0.009)
RI P ENING

var. ND 0.218 ND
Ponteroza [0.019]
var. Soldam ND 0.439 ND
[0.029]
Rosaceae Prunus spp. Cherry ND ND ND Lako et al.
— 0.14 (0.06) − (2007),
Bhaskarachary
et al. (1995)
P H Y SIOLOG Y

— 0.028 — Holden et al.

(1999)
var. Domestic 0.018 0.071 ND Yano et al.
(0.004) (0.004) (2005)
 var. USA ND 0.037 ND Yano et al.
(0.004) (2005)
Pyrus sp. Pear 0.006 0.027 ND Holden et al.
ROLE

(1999)
Rubus sp. Raspberry 0.012 0.008 — Holden et al.
O F

(1999)
Rutaceae Citrus aurantium L. Orange — 0.17 (0.08) — Bhaskarachary
et al. (1995)
maxima Pummelo 0.014 0.32 — Holden et al.
Merr. (1999)
microcarpa Musk lime — 0.012 — Tee and Lim
P IGMENTS

Bunge (1991)
nobilis L. Orange — 0.025 — Tee and Lim
paradisiaca Grapefruit ND 0.452 1.869 (1991)
Macfad. var. [0.019] [0.654] Yano et al.
Star ruby (2005)
var. Pink 0.005 0.603 — Holden et al.
D URING

[0.005] [0.152] (1999),

— — 3.36 Bramley (2000)
var. White 0.008 0.014 —
sinensis (L.) Orange 0.016 0.051 — Holden et al.
F RUIT

Osbeck (1999)
(Continued )
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267
268
Table 9.1  Carotenoid Contents (mg/100 g fresh weight) of Some Common Fruits
Family Genus Species Name α-Carotene β-Carotene Lycopene References
var. Navel 0.019 0.139 ND Yano et al.
[0.002] [0.014] (2005)
var. Valencia 0.015 0.051 ND Yano et al.
P OST H AR V EST

[0.001] [0.004] (2005)

reticulata Mandarin — 0.081 — Tee and Lim
Blanco (1991)
Orange ND 0.03 ND Lako et al.
(2007)
ND, not detected; —, data not available; var., variety; mean (standard deviation), mean [standard error].
RI P ENING
P H Y SIOLOG Y
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Table 9.2  Anthocyanin Contents of Some Common Fruits

Anthocyanin (mg/100 g
Fruit fresh weight) References
Red grape 30–750 Lamikanra (1989)
Apple, Red 1.7 Koponen et al. (2007)
Delicious
Blackberry 82.5–325.9 Torre and Barrit (1977), Wang
and Lin (2000)
Blueberry 25–495 Ballinge et al. (1972), Makus and
Ballinger (1973)
Bilberry 300–698 Mazza and Miniati (1993)
Cherry 2–450 Harbone and Hall (1964), Mazza
and Miniati (1993), Gao and
Mazza (1995)
Chokeberry 410–1480 Wu et al. (2006), Koponen et al.
(2007)
Cranberry 67–140 Wu et al. (2006), Koponen et al.
(2007)
Crowberry 360 Koponen et al. (2007)
Gooseberry 2.0–43.3 Wu et al. (2006), Pantelidis et al.
(2007)
Grapefruit 5.9 Koponen et al. (2007)
Peach 4.2 Koponen et al. (2007)
Pear 5–10 Mazza and Miniati (1993)
Pomegranate 600–765 Kriventsov and Arendt (1981)
(juice)
Raspberry 20–687 Wang and Lin (2000), Wu et al.
(2006)
Red apple 1.3–12 Mazza and Velioglu (1992),
Wu et al. (2006)
Rowanberry 14 Koponen et al. (2007)
Saskatoon 234 Koponen et al. (2007)
berry
Strawberry 19–55 Lopes-da-Silva et al. (2007)

as flavonoids (Pierantoni et al., 2010). Anthocyanin biosynthesis has been

well characterized in flowers of petunia (Petunia hybrida) and snapdragon
(Antirrhinum majus) and in kernels of maize (Zea mays) (Holton and
Cornish, 1995), with the anthocyanin biosynthetic pathway being one of the
well-known pathways in plants (Dixon and Steele, 1999). The precursors

269
P OST H AR V EST RI P ENING P H Y SIOLOG Y

for the synthesis of anthocyanin are malonyl-CoA and 4-coumaroyl-CoA,

which is condensed to chalcone by chalcone synthase (CHS). Chalcone
isomerase (CHI) then catalyzes the isomerization of chalcone to the flava-
none. Flavanone 3-hydroxylase (F3H) catalyzes the hydroxylation of fla-
vanones at the 3 position of the C-ring to form the dihydroflavonols, which
are then converted to leucoanthocyanidins by dihydroflavonol 4-reductase
(DFR). Anthocyanidin synthase (ANS) converts leucoanthocyanidins to
anthocyanidins, the first colored compound in the anthocyanin pathway.
The final step in the anthocyanin pathway is 3-glycoside formation by
UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT). In apple, UDP-
galactose:flavonoid 3-O-galactosyltransferase (UFGalT) might be involved
in this step since cyanidin 3-galactoside is the major pigment in the red
skin of apple (Lancaster, 1992). Regulatory genes that control expression
of the structural genes in the biosynthetic pathway have also been identi-
fied in many plants. These genes influence the intensity and pattern of the
expression of many different anthocyanin biosynthesis genes (Holton and
Cornish, 1995). Anthocyanin biosynthesis in grape skin has been exten-
sively studied. It has been determined that anthocyanins are synthesized
from phenylalanine through an anthocyanin biosynthetic pathway regu-
lated by enzyme activities (Hrazdina et al., 1984) and gene expressions
(Boss et al., 1996). Some anthocyanin biosynthetic pathway genes, includ-
ing a large number of isogenes—phenylalanine ammonia lyase (PAL), chal-
cone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase
(F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygen-
ase (LDOX), UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT), and
so on—have been cloned (Sparvoli et al., 1994; Mori et al., 2005) (Table 9.3).

9.5  Grape as a Model System for

Pigmentation Studies in Fruit Crops
Grapevine (Vitis vinifera L.) is the most important fruit crop and has been
widely cultivated in the world, with a long history of physiological, bio-
chemical, and molecular investigations and advances in the development
of genetic and molecular tool kits for this species. The grapevine species
is a representative of the genus Vitis and has a disomic inheritance with
2n = 38 and a relatively small genome of 475 Mbp (Lodhi and Reisch, 1995;
Aradhya et al., 2003). Grape skin color is an important factor both biologi-
cally and economically, since the quality of grape berries greatly depends
on skin color, which greatly affects market prices and the wine industry
(Koshita et al., 2011). Fruit skins of grapes have the highest amounts of
volatile and polyphenolic compounds compared to other parts of the fruit.
The color of grape berries is determined by the quantity and composition of

270
 Table 9.3  Regulatory Genes Identified in Different Fruit Crops and Their Targeting Structural Genes Involved in Anthocyanin
Biosynthesis
Structural
ROLE

Fruit Species MYB bHLH Genes Fruit Tissue Accession No. References
Grapes VvMYB5a UFGT, C4H, Berry skin AY555190 Deluc et al. (2006)
O F

ANS, F3H
VvMYB5b MYCA1 VvLAR, VvANR Berry skin AY899404 Deluc et al. (2008)
VvMYBA1 CHI, CHS, Berry skin DQ886418 Walker et al. (2007)
4CL, DFR
VvMYBA2 UFGT, ANR DQ886419 Walker et al. (2007)
VlmybA1-1 UFGT AB073010 Kobayashi et al.
P IGMENTS

(2002)
VlmybA1-2 UFGT, OMT, AB073012 Kobayashi et al.
GST (2002)
VlmybA2 UFGT AB073013 Kobayashi et al.
(2002)
D URING

VlmybB1-1 Unknown AB073016 Kobayashi et al.

(2002)
VlmybB1-2 Unknown AB073017 Kobayashi et al.
(2002)
F RUIT

VlmybC Unknown AB073014 Kobayashi et al.

(2002)
VlmybD Unknown AB073015 Kobayashi et al.
VvmybA1 UFGT AB097923 (2002, 2004)
(Continued )
RI P ENING

271
272
Table 9.3  Regulatory Genes Identified in Different Fruit Crops and Their Targeting Structural Genes Involved in Anthocyanin
Biosynthesis
Structural
Fruit Species MYB bHLH Genes Fruit Tissue Accession No. References
VvmybA2 UFGT AB097924 Kobayashi et al.
P OST H AR V EST

(2004)
VvmybA3 Unknown AB097925 Kobayashi et al.
(2004)
VvMybA1 UFGT DQ886417 Walker et al. (2007)
VvMybA2r UFGT DQ886419 Walker et al. (2007)
VvMybA2w Unknown DQ886420 Walker et al. (2007)
VvMybA3 Unknown DQ886421 Walker et al. (2007)
RI P ENING

VvMybPA1 CHI, F3′ 5′ H, AM259485 Bogs et al. (2007)

ANR, LAR
Apple MdMYB1 MdHLH3 ANS Peel DQ886414 Takos et al. (2006)
MdMYBA HLH33 Peel AB279598 Ban et al. (2007)
MdMYB10 Peel/flesh/ EU518250 Espley et al. (2009)
leaf
Strawberry FaMYB1 ANS, UFGT Flesh AF401220 Aharoni et al. (2001)
P H Y SIOLOG Y

FaMYB10 DFR EU155162 Wang et al. (2010)

FvMYB10 DFR EU155163 Wang et al. (2010)
Pear PyMYB10 DFR Peel GU253310 Feng et al. (2010)
PcMYB10 Peel HM775223 Pierantoni et al.
PbMYB10 EU153577 (2010), Wang et al.
(2010)
 PpyMYB10 EU153576 Wang et al. (2010)
Peach PprMYB10 DFR Unknown EU155160 Wang et al. (2010)
ROLE

Bilberry VmTDR4 VmMYB2 Peel/flesh FJ418852 Jaakola et al. (2010)

Red raspberry RiMYB10 DFR Unknown EU155165 Wang et al. (2010)
O F

Chinese MrMYB1 AtDFR Flesh GQ340767 Niu et al. (2010)

bayberry
Mangosteen GmMYB10 GmDFR, Pericarp FJ197137 Palapol et al. (2009)
GmUFGT,
GmLDOX
Loquat EjMYB10 Unknown EU153572 Wang et al. (2010)
Quince CoMYB10 Unknown EU153571 Wang et al. (2010)
P IGMENTS

Apricot ParMYB10 DFR Unknown EU153178 Wang et al. (2010)

Cherry PavMYB10 DFR Unknown EU153581 Wang et al. (2010)
PcrMYB10 DFR Unknown EU153582 Wang et al. (2010)
PcfMYB10 Unknown EU153583 Wang et al. (2010)
Almond PdMYB10 DFR Unknown EU155159 Wang et al. (2010)
D URING

Plum PiMYB10 Unknown EU153579 Wang et al. (2010)

PdmMYB10 Unknown EU153580
PsMYB10 Unknown EU155161 Wang et al. (2010)
F RUIT
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273
P OST H AR V EST RI P ENING P H Y SIOLOG Y

anthocyanins in their skins (Pomar et al., 2005; Torres et al., 2010). Though
earlier studies were focused more on the molecular biology of fruit skin
color, genomics approaches have recently been used to reveal insights
into the control of primary coloration upstream of anthocyanin, coloration-
related signal transduction systems, and downstream metabolic networks.
Grape pleiotropic coloration mutations have added greatly to the under-
standing of fruit skin color. Genes of anthocyanin production and pigment
biosynthesis enzymes were among the first anthocyanin-responsive genes
isolated from fruits of grape, and anthocyanin biosynthesis in grape skin
has been extensively studied (Boss et al. 1996). In grapevine, anthocyanin
biosynthesis depends on Myb-related genes, which play a crucial role in
the different varieties that exhibit an assortment of biosynthesis modes. In
American hybrid grapevine, VlmybA1-2, VlmybA2, and VlmybA3 are located
in the pericarp, though the exact location is unknown, and also regulate
the anthocyanin biosynthetic gene cluster (dashed box in Figure 9.2). Gene
clusters can encode the production of the Myb protein, thus contributing to
the expression of the anthocyanin biosynthetic pathway in UFGT-turned
grape skin color. Grape color loci are mainly divided into three cases
(Figure 9.2):

1. VvmybA1 and VvmybA2 can be mutated, VvmybA1a and VvmybA2w

have no function, and VvmybA1b, VvmybA1c, and VvmybA2r have a
function. In addition, some red-white mutant varieties, VvmybA1
and VvmybA2, are lost and cannot synthesize anthocyanins, though
VvmybA2r and VvmybA1c, VvmybA2w and VvmybA1b, and VvmybA2w
and VvmybA1c can synthesize anthocyanins in Europe and American
hybrid grapes.
2. The European oriental species has only VvmybA1SUB and lacks
VvmybA2, VlmybA1-2, VlmybA2, and VlmybA3. Whether VvmybA1SUB
can regulate anthocyanin biosynthesis in grapevine is still unknown
(dashed arrows in Figure 9.2).
3. In European and American hybrid grapes, the VlmybA1-3 gene
cluster can be encoded separately, and VlmybA1-2 and VlmybA2
composition generating the Myb protein induced UFGT to express
anthocyanins in the skin.

Due to the research importance and the economic value of Arabidopsis

t­ haliana (chromosome no. = 5) and V. vinifera (chromosome no. = 19), these
two model plant species have been widely used for investigation of the struc-
tural genes involved in the anthocyanin biosynthesis pathway. These genes
include phenylalanine ammonia lyase (PAL), cinnamate 4-­ hydroxylase
(C4H), 4-­coumarate-CoA ligase (4CL), chalcone synthase (CHS), chalcone
isomerase (CHI), flavanone 3-hydroxylase (F3H), flavanone 3′-hydroxylase

274
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Chr2
VvmybA2 VvmybA1

Mutation
I
Regulatory
function in red/
black pericarp
Chr2
Chr2
VvmybA2w VvmybA1a VvmybA2r VvmybA1c VvmybA1SUB
Chr2 VvmybA2w VvmybA1b
null null
VvmybA2w VvmybA1c II

No regulatory Coding sequence

MYB Promoter
function in
UFGT
white pericarp

Anthocyanin
Delphinidin Chr2
UFGT III VvmybA1-2 VvmybA1-3
Chr2
VvmybA2 VvmybA1-3

Anthocyanin OMT

Regulatory
function in red/
black pericarp

Figure 9.2  Schematic diagrams of the regulation of the skin color locus of
grapevine.

(F3′H), flavanone 3′,5′-hydroxylase (F3′ 5′ H), dihydroflavonol reductase

(DFR), anthocyanidin synthase/leucoanthocyanidin dioxygenase (ANS/
LDOX), and UDP-flavonoid glucosyltransferase (UFGT). After blast search
and protein domain analysis, the homologous genes involved in the antho-
cyanin biosynthesis pathway were isolated (Table 9.4). PAL is the first
structural gene in the pathway, and four copies (AtPAL1, AtPAl2, AtPAL3,
and AtPAL4) have been reported in A. thaliana and 11 in V. vinifera. AtC4H
is located in chromosome 2 of A. thaliana, and three copies of C4H have
been isolated in V. vinifera, with two copies of C4H being in chromosomes
6 and  11. Vv4CL5 was located in chromosome 1_random, while VvC4H3,
Vv4CL8, and VvCH3 were linked to chromosome unknown_random. In
total, 11 grapevine structural genes (18.18%) were located in random
sequences, with 8 being located in chromosome unknown_­random, 2 in
chromosome 16_random, and 1 in chromosome 1_random.

275
276
Table 9.4  Homolog Genes Involved in Anthocyanin Biosynthesis in Arabidopsis and Grape
Arabidopsis
Gene Gene ID Chromosome Start End Grape Gene Gene ID Chromosome Start End
AtPAL1 AT2G37040 2 15,557,376 15,560,363 VvPAL1 GSVIVP00013939001 chr16 700,746 703,220
AtPAL2 AT3G53260 3 19,744,051 19,746,780 VvPAL2 GSVIVP00013930001 chr16 648,662 651,184
AtPAL4 AT3G10340 3 3,204,013 3,208,022 VvPAL3 GSVIVP00013943001 chr16 724,118 727,239
P OST H AR V EST

AtPAL3 AT5G04230 5 1,160,831 1,163,775 VvPAL4 GSVIVP00013928001 chr16 618,476 621,190

VvPAL5 GSVIVP00013947001 chr16 779,795 782,254
VvPAL6 GSVIVP00013922001 chr16 597,226 599,843
VvPAL7 GSVIVP00023211001 chr8 11,718,432 11,721,087
VvPAL8 GSVIVP00018175001 chr13 5,969,524 5,991,084
VvPAL9 GSVIVP00024561001 chr6 3,222,841 3,226,347
RI P ENING

VvPAL10 GSVIVP00013927001 chr16 612,547 614,853

VvPAL11 GSVIVP00013924001 chr16 606,754 608,798
AtC4H AT2G30490 2 12,993,663 12,995,770 VvC4H1 GSVIVP00023932001 chr6 8,813,071 8,816,574
VvC4H2 GSVIVP00017017001 chr11 10,158,634 10,160,529
VvC4H3 GSVIVP00007155001 chrUn_random 45,548,882 45,550,610
At4CL1 AT1G51680 1 19,158,752 19,161,552 Vv4CL1 GSVIVP00031383001 chr11 12,980,141 12,983,730
At4CL2 AT3G21240 3 7,454,269 7,457,379 Vv4CL2 GSVIVP00031385001 chr11 13,004,990 13,010,232
At4CL3 AT1G65060 1 24,167,202 24,171,502 Vv4CL3 GSVIVP00014031001 chr16 2,123,569 2,128,535
At4CL5 AT3G21230 3 7,448,040 7,452,000 Vv4CL4 GSVIVP00022179001 chr6 22,071,551 22,079,236
P H Y SIOLOG Y

At4CL4 AT5G63380 5 25,387,411 25,390,063 Vv4CL5 GSVIVP00002799001 chr1_random 48,50,822 4,853,094

At4CL6 AT4G05160 4 2,664,385 2,666,707 Vv4CL6 GSVIVP00018101001 chr13 7,219,492 7,221,432
At4CL7 AT4G19010 4 10,411,490 10,414,249 Vv4CL7 GSVIVP00015553001 chr18 1,226,323 1,230,351
At4CL8 AT5G38120 5 15,213,765 15,216,205 Vv4CL8 GSVIVP00003591001 chrUn_random 26,173,966 26,179,603
At4CL9 AT1G20510 1 7,103,454 7,105,881 Vv4CL9 GSVIVP00020555001 chr14 15,331,127 15,333,537
 At4CL10 AT1G62940 1 23,310,535 23,312,747 Vv4CL10 GSVIVP00020554001 chr14 15,323,828 15,330,405
At4CL12 AT1G20500 1 7,100,502 7,102,915
At4CL14 AT1G20480 1 7,094,833 7,097,114
ROLE

AtCHS AT5G13930 5 4,488,688 4,490,264 VvCHS1 GSVIVP00037969001 chr14 13,875,792 13,877,198

VvCHS2 GSVIVP00037967001 chr14 13,889,324 13,890,882
O F

VvCHS3 GSVIVP00006341001 chrUn_random 37,798,575 37,800,731

AtCHI AT3G55120 3 20,430,114 20,431,470 VvCHI GSVIVP00029513001 chr13 2,128,014 2,129,693
AtF3H AT3G51240 3 19,025,264 19,026,939 VvF3H1 GSVIVP00036784001 chr4 14,790,912 14,792,727
VvF3H2 GSVIVP00014419001 chr18 12,160,108 12,162,146
VvF3H3 GSVIVP00036782001 chr4 14,832,092 14,835,848
AtF3′ H AT5G07990 5 2,560,394 2,563,109 VvF3′ H1 GSVIVP00016217001 chr17 8,112,032 8,114,427
VvF3′ H2 GSVIVP00016215001 chr17 8,135,555 8,137,842
P IGMENTS

AtF3′ 5′ H1 AT4G12300 4 7,307,737 7,309,755 VvF3′ 5′ H1 GSVIVP00007272001 chr2 4,774,838 4,777,086

AtF3′ 5′ H2 AT4G12320 4 7,314,778 7,316,669 VvF3′ 5′ H2 GSVIVP00007269001 chr2 4,754,383 4,756,418
AtF3′ 5′ H3 AT4G12310 4 7,310,416 7,312,577 VvF3′ 5′ H3 GSVIVP00010339001 chrUn_random 63,649,713 63,651,747
AtF3′ 5′ H4 AT4G12330 4 7,317,560 7,319,739 VvF3′ 5′ H4 GSVIVP00007267001 chr2 4,745,715 47,47,542
AtF3′ 5′ H5 AT5G44620 5 17,997,779 17,999,558 VvF3′ 5′ H5 GSVIVP00010337001 chrUn_random 63,642,459 63,644,286
D URING

AtF3′ 5′ H6 AT4G22690 4 11,929,359 11,931,693 VvF3′ 5′ H6 GSVIVP00010332001 chrUn_random 63,611,190 63,613,352

AtF3′ 5′ H7 AT4G22710 4 11,934,969 11,936,755 VvF3′ 5′ H7 GSVIVP00010336001 chrUn_random 63,635,888 63,637,834
VvF3′ 5′ H8 GSVIVP00007266001 chr2 4,738,982 4,741,051
VvF3′ 5′ H9 GSVIVP00038447001 chr16_random 1,531,571 1,533,743
F RUIT

VvF3′ 5′ H10 GSVIVP00038441001 chr16_random 1,444,657 1,446,826

VvF3′ 5′H11 GSVIVP00038443001 chr16_random 1,473,707 1,475,916
VvF3′ 5′H12 GSVIVP00001045001 chr2 4,336,836 4,339,311
VvF3′ 5′ H13 GSVIVP00026276001 chr15 6,313,161 6,315,563
RI P ENING

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

9.6  Case Studies

9.6.1 Apple
Apple (Malus spp.) is one of the most popular fruit crops worldwide. There
are more than 7500 known cultivars of apples, with different cultivars
available for temperate and subtropical climates. Among the quality char-
acteristics of the cultivars, red fruit skin coloration is of major importance
and significantly determines the market value of the produce, because
consumers associate better-colored apples with better taste, ripeness, and
flavor (King and Cliff, 2002). Color is also an obvious characteristic that
facilitates product discrimination. For example, the apple cultivar ‘Cripps’
Pink’, commercially sold as Pink Lady, has been very successful due to
its distinct pink hue (Corrigan et al., 1997). The coloration of apple skin is
derived from anthocyanins belonging to a class of flavonoids (Gallus et al.,
2005; Liu et al., 2005; Rasmussen et al., 2005). Anthocyanin ­accumulation
positively correlates with the expression of at least five anthocyanin
biosynthetic genes (MdCHS, MdF3H, pDFR, MdANS, and pUFGluT ),
­
­suggesting that the levels for the expression of the five genes basically cor-
respond to the degree of anthocyanin concentration (Honda et al., 2002;
Ubi et al., 2006).
Apple skin is also a rich source of compounds that are potent antioxi-
dants, such as anthocyanins, chitosans (CTs), and flavonols, whose intake
in the human diet has been correlated with lower incidences of cancer
and cardiac disease (Gallus et al., 2005; Liu et al., 2005; Rasmussen et al.,
2005). For these reasons, the antioxidative potential of apple skin may
become a new quality marker for different apple cultivars (Schirrmacher
and Schempp, 2003). Although the pigments that comprise fruit color vary
with fruits, the amount and composition of anthocyanins that belong to a
class of flavonoids are the major determinants of apple fruit skin redden-
ing. Anthocyanin biosynthesis in apple fruit is developmentally regulated
and occurs at two peaks. The first peak occurs at the fruitlet stage in both
red and nonred cultivars, and this peak is not economically important. The
second peak takes place later, at the ripening fruit stage, only in red cul-
tivars. The intensity and extent of reddening at the mature fruit stage are
most important, as they impact greatly the marketable value of the produce
and have direct bearing in the economics of apple production. The expres-
sion of the five anthocyanin biosynthetic genes, chalcone synthase (CHS),
flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocy-
anidin synthase (ANS), and UDP-glucose:flavonoid 3-O-glucosyltransferase
(UFGluT) during ripening in five early ripening apple cultivars, as well as
under UV-B and temperature treatments. The expression of CHS, ANS and
UFGluT was substantially depressed in fruit skin, whereas that of all five

278
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

genes including CHS, ANS and UFGluT was enhanced by UV-B and low
temperature treatments with the accumulation of anthocyanins. The accu-
mulation level of the main anthocyanin pigment, cyanidin 3-galactoside,
in the fruits under UV-B and low temperature treatment, was less than
that in the field-ripened fruits, while cyanidin 3-arabinoside was relatively
the same level. The UV-B and low temperature were important factors for
anthocyanin accumulation in apple fruit skin by inducing the expression
of the anthocyanin biosynthetic genes, especially CHS, ANS and UFGluT
genes. The recent studies at the molecular level have revealed that the
regulation of the expression of the biosynthetic genes is one of the causes
that determine the anthocyanin accumulation in apple skin.

9.6.2 Strawberry
Strawberry (Fragaria spp.) fruit is widely appreciated, not only for its
characteristic aroma, but also for its bright red color. Besides sweetness
and flavor, the development of red color is one of the most important char-
acteristics affecting the fruit quality of strawberry, as it contributes to
a consumer’s initial impression and decision to buy or not. These fruit
offer an interesting model to study fruit pigmentation by anthocyanin,
considering that the color of the white and red botanical forms reflects
the presence of such phytochemicals. In the Fragaria genus, fruit color
is determined by the accumulation of anthocyanin, the most abundant
flavonoid in strawberry fruits (Hannum, 2004). Pelargonidin 3-gluco-
side is the predominant anthocyanin in several varieties of red straw-
berries, usually followed by pelargonidin 3-rutinoside and cyanidin
3-­glucoside (Gil et al., 1997; Kosar et al., 2004; Tulipani et al., 2008).
These three compounds represent more than 95% of the total anthocya-
nins in the cultivated strawberry (Lopes-da-Silva et al., 2007). Flavonoid
genes involved in anthocyanin biosynthesis exhibit upregulation dur-
ing  ­ripening, leading to fruit pigmentation in strawberry and thereby
establishing a positive correlation between transcript levels of flavonoid
genes and anthocyanin accumulation (Manning, 1998; Almeida et al.,
2007; Carbone et al., 2009). The cDNA fragments of genes from phenyl-
propanoid (PAL, C4H, and 4CL) and the flavonoid biosynthetic pathway
(CHS, CHI, F3H, DFR, ANS, UFGT, LAR, and ANR) have been isolated
from the native Chilean strawberry, and the differential transcriptional
profiles of these genes analyzed in Fragaria chiloensis and Fragaria
patagonica through four different fruit developmental stages and tis-
sues by quantitative real-time polymerase chain reaction (qRT-PCR).
In addition, the accumulation of the two major anthocyanins, pelargo-
nidin 3-glucoside and cyanidin 3-glucoside, in these native strawberries

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

was quantified by high-performance liquid chromatography with diode

array detection (HPLC-DAD). Salvatierra et al. (2010) discussed the
relationship between transcriptional profiles and anthocyanin contents
present in terms of the modulation of flavonoid gene expression in the
determination of the different pigmentation patterns observed in the
white- and red-fruited Chilean native strawberries.

9.6.3 Tomato
Tomato (Lycopersicon esculentum) has dark green foliage and elevated
anthocyanin expression in the hypocotyls of seedlings and in the skin
and outer pericarp tissues of fruits. Tomato fruit is not usually reported
to contain anthocyanin (Jones et al., 2003), but Oregon State University
managed to develop a high-anthocyanin tomato cultivar using genes intro-
gressed from wild species. Cultivated tomatoes produce anthocyanin in
vegetative tissues, but only small amounts of other flavonoids, such as nar-
ingenin chalcone and flavonols, can be found in the fruit (Muir et al., 2001;
Torres et al., 2005; Bovy et al., 2007). As a consequence, tomato is consid-
ered an excellent candidate for an enhancement of the flavonoid and antho-
cyanin contents through transgenic approaches (Gonzali et  al., 2009).
Recently, Butelli et al. (2008) expressed in tomato Delila and Rosea1, two
genes coding for transcription factors involved in anthocyanin produc-
tion in snapdragon. The tomato anthocyanin fruit (Aft) genotype is char-
acterized by purple color in the skin and outer pericarp of its fruits due
to higher levels of anthocyanins—flavonoid metabolites. The dominant
gene Aft (Anthocyanin fruit) was introgressed into domesticated tomato
plants by a cross with Solanum chilense (Jones et al., 2003). Aft triggers
anthocyanin accumulation in immature green fruit upon stimulation by
high light. Subsequently, pigments are produced continuously through-
out development (Mes et al., 2008). The Aft gene identity still has to be
revealed. Recent linkage analyses showed that the Aft locus cosegregates
with two different MYB transcription factor genes located on chromo-
some 10, SlAN2 (Mes et al., 2008; Boches et al., 2009) and anthocyanin
1 (SlANT1) (Sapir et al., 2008), both involved in anthocyanin synthesis
in tomato (Mathews et al., 2003; Mes et al., 2008). A recessive gene, Atv
(Atroviolacea), derived from the interspecific cross with Solanum chees-
maniae (L. Riley) Fosberg, has been shown to influence anthocyanin pig-
mentation in the entire tomato plant, particularly in the vegetative tissues
(Mes et al., 2008). The Atv gene has been located to chromosome 7 (Rick
et al., 1968), and previous studies indicated that its mutation may affect
phytochrome responses, since Atv plants exhibit an exaggerated response
to red light in terms of anthocyanin production (Kendrick et al., 1997). The

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anthocyanidin profile of Aft/Aft Atv/Atv fruit was consistent with results

from previous analyses on high-­a nthocyanin tomato (Jones et al., 2003;
Mathews et al., 2003), indicating that Aft and Atv alleles likely do not affect
the structural genes of the biosynthetic pathway. No influence of anthocy-
anin accumulation on carotenoid levels was detected (Mes et al., 2008).
Povero et al. (2011) published a detailed transcript profiling analysis of
the anthocyanin biosynthetic pathway that was carried out in these tomato
fruit and revealed sets of differentially regulated genes and synergistic
effects of Aft and Atv. They demonstrate that the regulation of transcripts
involved in phenylpropanoid metabolism is tightly linked to the anthocy-
anic phenotypes of tomato fruit and is also accompanied by changes in
other structural and metabolic traits.

9.6.4 Litchi
Litchi (Litchi chinensis Sonn.) is a tropical and subtropical fruit that has
high commercial value due in part to its white and translucent aril and
attractive red color (Holcroft and Mitcham, 1996; Jiang et al., 2004).
The red color of the litchi fruit peel is mainly attributed to anthocyanin
(Prasad and Jha, 1978; Jiang et al., 2006). Litchi fruit pericarp contains
a large amount of pigments that are responsible for the red color (Lee
and Wicker, 1991). Cyanidin was identified as the major anthocyanin
present in pericarp tissues of fresh litchi fruit, followed by delphinidin
and pelargonidin (Oomah and Mazza, 1999; Jiang et al., 2004) and mal-
vidin (Prasad and Jha, 1978; Lee and Wicker, 1991). Pericarp browning
is a common and important defect of harvested litchi fruit, which results
in reduced market value (Huang and Scott, 1985) and has mainly been
attributed to the degradation of anthocyanin and the oxidation of pheno-
lics by polyphenol oxidase (PPO) (Jiang et al., 2004). Thus, postharvest
technologies have been developed to reduce the anthocyanin degradation
and maintain the red color of harvested litchi fruit (Jiang et al., 2006).
Lee and Wicker (1991) identified the red pigment in litchi as cyanidin
3-rutiside. Cyanindin 3-glucoside and malvidin 3-glucoside may also be
present in these anthocyanins (Zhang et al., 2000). Sarni-Manchado et al.
(2000) identified the anthocyanins as cyanidin 3-rutinoside, cyanidin glu-
coside, quercetin 3-rutinoside, and quercetin glucoside, while Zhang et al.
(2004) reported that the major anthocyanin of litchi fruit pericarp was
cyanidin 3-rutinoside. The differences in the reported identification of the
anthocyanins may be attributed to differences in the extraction and puri-
fication procedures (Jiang et al., 2004). Most of the red pigments in litchi
fruit pericarp present in the ethyl acetate fraction and the pigments are
identified as flavanols. Identification of structures of flavanols was needed

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to explain the loss in the skin color of harvested litchi fruit, in relation to
their antioxidant activity.
Recent research has shown that pigments in postharvest fruits
exhibit a strong antioxidant activity (Einbond et al., 2004; Bao et al., 2005).
Antioxidative properties of pigments resulted from their high reactivity
as hydrogen or electron donors and from their ability to chelate transition
metal ions, that is, terminate the Fenton reaction (Rice-Evans et al., 1997;
Wada and Ou, 2002). It is well established that enzymatic browning of post-
harvest fruits is related to antioxidant activity (Martinez and Whitaker,
1995). Unfortunately, not much information on the antioxidant activ-
ity of skin pigments from harvested litchi fruit, as well as the ­molecular
­mechanisms regulating this activity, is available.

9.6.5  Chinese Bayberry

The Chinese bayberry (Myrica rubra Sieb. and Zucc.) is an important eco-
nomic Asian fruit crop belonging to the Myricaceae family (Chen et  al.,
2004). This is a fruit where red color is one of the key fruit quality traits.
Anthocyanins are the principal color compounds in this species, with
more than 85% of fruit anthocyanins present as cyaniding 3-O-glucoside,
and no delphinidin, malvidin, and petunidin anthocyanins found present
(Zhang et al., 2008; Fang et al., 2009). This species provides an interest-
ing model for dissecting the causes of color variability in fruit, since a
range of cultivars exists, producing fruit from white (no obvious anthocy-
anin accumulation, e.g., the cultivar ‘Shuijing’) through red (‘Dongkui’) to
dark red-purple (‘Biqi’) (Zhang et al., 2009). Genes encoding the antho-
cyanin biosynthesis enzymes chalcone synthase, chalcone isomerase,
flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), dihy-
droflavonol 4-reductase (DFR), anthocyanidin synthase (ANS) and UDP-
glucose:flavonoid 3-O-glucosyltransferase (UFGT), as well asMrMYB1, a
R2R3 MYB transcription factor homologous to known activators of antho-
cyanin biosynthesis, were isolated from ripe fruit of BQ (Niu et al., 2010).
Differences in mRNA abundance of MrF3H, MrF3’H, MrDFR1, MrANS,
and MrUFGT highly correlated with differential accumulation of anthocya-
nins between cultivars, suggesting coordinated regulation by transcription
factors. The transcript level of MrMYB1 was strongly associated with the
anthocyanin content in ripe fruit. Overexpression of MrMYB1 both stimu-
lated anthocyanin accumulation and activated an  Arabidopsis-DFR  pro-
moter in tobacco. MrMYB1d, an allele with a 1 bp deletion at nucleotide 30
of the coding sequence, suggests that a nonsense mutation of the MYB1
protein may be responsible for no or low expression of MYB1 in the white
and red fruit.

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9.6.6  ‘Hass’ Avocados

‘Hass’ avocado fruit color change is important as an indicator of ripeness
for both industry and consumers. Cox et al. (2004) sought to character-
ize pigment changes during ripening of ‘Hass’ avocados and observed that
Cox et al. (2004) sought to characterize pigment changes during ripen-
ing of ‘Hass’ avocados and observed that skin color and changes in chlo-
rophyll and anthocyanin concentrations in relation to fruit firmness and
ripening temperature. As fruit ripened,  skin color  changed from green
to purple to black, which was observed as a decrease in Lightness (L),
chroma, and hue. Total anthocyanins in skin tissue increased during rip-
ening, but this increase is due almost entirely to a single anthocyanin:
cyanidin 3-O-glucoside. Cyanidin 3-O-glucoside concentration increased
after 3–6 days postharvest (depending on ripening temperature), and lev-
els increased earlier, and are higher, with higher ripening temperature.
Chlorophyll a and b levels decreased until 4–5 days after harvest, but did
not change significantly thereafter. No significant differences in chlo-
rophyll concentration are observed among the three ripening tempera-
tures. Fruit harvested directly from the tree (unripe) with skin darkening
showed pigment changes similar to those during ripening (reduced chlo-
rophyll and increased cyanidin 3-O-glucoside). Color change in ‘Hass’ avo-
cados from green to purple and then black results from an initial decrease
in chlorophyll content, followed by an increase in the levels of the anthocy-
anin cyanidin 3-O-glucoside.

9.6.7 Pear
Pear fruit  pigmentation  (red and green pear) is observed through tran-
scriptional and chemical approaches. The proportion of different anthoy-
anins is demonstrated to be characteristic of each botanical form, with
pelargonidin 3-glucoside being the most abundant in red fruit and cyani-
din 3-glucoside the major one in green fruit. Partial gene sequences of
the phenylpropanoid and flavonoid  biosynthesis pathways are isolated
from the native Chilean strawberry fruits and used to design gene-spe-
cific primers in order to perform transcriptional analyses by qRT-PCR.
These genes show spatial, developmental, and genotypic-associated pat-
terns. The red fruit exhibited higher transcript levels of anthocyanin-
related genes and higher levels of anthocyanin than the green pigmented
fruit. The anthocyanin accumulation in red and green fruits is concomi-
tant with the particular progress of the transcriptional activity of genes
involved in the  biosynthesis  of flavonoid pigments. The differences in
anthocyanin contents, in terms of both type and quantity, between the two

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botanical forms are coincident with the differential transcriptional pat-

terns found in the anthocyanin-related genes.

9.6.8 Cherry
The cherry (Prunus cerasus L.), containing high levels of anthocyanins that
possess strong antioxidant and anti-inflammatory properties, has attracted
much attention. The evaluation and characterization of the in  vitro–­
produced pigments are compared to those of the anthocyanins found
in vivo in fruits of several cherry cultivars. Interestingly, the anthocyanin
profiles found in whole fruit extracts are similar in all tested genotypes.
The evaluation of antioxidant activity, performed by oxygen radical absor-
bance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC)
assays, revealed a relatively high antioxidant capacity for the fruit extracts
and a lower one for the callus extract. The structural gene DFR was deter-
mined from cherry where two MYB factors were responsible: PavMYB10
and PcrMYB10 (Wang et al., 2010).

9.6.9 Kiwifruit
The kiwifruit or Chinese gooseberry (often shortened to kiwi) is the edible
berry of a woody vine in the genus Actinidia, and it is native to northern
China. The anthocyanins responsible for the red color of red kiwifruit
were extracted in acidified ethanol and isolated by solid-phase extraction
(SPE), followed by preparative HPLC. Five anthocyanins were obtained
and subsequently identified as delphinidin 3-[2-(xylosyl)galactoside],
delphinidin 3-galactoside, cyanidin 3-[2-(xylosyl)galactoside], cyanidin
3-galactoside, and cyanidin 3-glucoside by a combination of liquid
chromatography–mass spectrometry (LC-MS)/ mass Spectrometry (MS),
Gas chromatography–mass spectrometry (GC-MS), and Two-dimensional
nuclear magnetic resonance spectroscopy (2D NMR).

9.7  Pigments from Fruit to Human Health

In recent years, studies have begun to investigate the specific properties
of isolated anthocyanin pigments; however, many still study the health
effects of anthocyanins from fruit extracts where anthocyanins are pres-
ent in combination with other compounds. In fact, some reports suggest
that anthocyanin activity is actually potentiated when delivered in mix-
tures, as opposed to isolates. Anthocyanins act as powerful antioxidants
and water-soluble vacuolar pigments, and also possess a multitude of

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biological roles, including protection against solar exposure and ultravio-

let radiation, free radical scavenging and antioxidative capacity, defense
against many different pathogens, attraction of predators for seed disper-
sal, and the newly proposed modulation of signaling cascades (Bornsk
et al., 2012).
There are still several aspects of anthocyanins’ pharmacological
roles that have remained elusive to scientists. In most of the interventions
of anthocyanins in human health, details on the mechanisms of action for
bioactivity, uptake, absorption, bioavailability, whole-body distribution, and
tissue localization are still not fully understood.
Pigments are able to prevent oxidative damage to DNA, proteins, lip-
ids, and other macromolecules caused by reactive oxygen species (ROS).
They have systemic action, since they are absorbed and circulate in the
blood, and it is this circulating form that acts upon different target tissues
in the human body. Finally, they may also act as topical agents, for example,
by protecting the skin from UV radiation. Free radical damage contributes
to the etiology of many chronic diseases (Bobe et al., 2006; Chen et al.,
2006; Valcheva-Kuzmanova and Belcheva, 2007), and thus antioxidants
may have beneficial effects on human health at different levels (visual acu-
ity, cancer, heart disease, age-related neurodegenerative disorders, etc.).
In research trials, anthocyanins have demonstrated the ability to reduce
cancer cell proliferation (growth and multiplication) and inhibit tumor for-
mation. The capacity of anthocyanin pigments to interfere with the pro-
cess of carcinogenesis seems to be linked to multiple potential mechanisms
of action, including the inhibition of cyclooxygenase (COX) enzymes and
potent antioxidant potential.

9.8  Transgenic Plants Developed for

Fruit Color
Transgenic fruit plants have been developed either to understand the
mechanism of fruit skin color or for use in biotechnological applications
and the fruit industry. Since the production of the first transgenic plants
in the 1980s, a wide diversity of patents have been sought, and granted, on
all aspects of the process, ranging from the underlying methods for tissue
culture to the means of introducing the heterologous DNA to the composi-
tion of the DNA construct introduced (Dunwell, 2005; Pray and Naseem,
2007). The amount of patent information available in the area of plant
transformation is substantial. A search of the U.S. application database
(http://www.bios.net/daisy/bios/patentlens.html) alone for “transgenic
plant” and “method” returns extensive results. Summaries of relevant
recently granted patents and patent applications in the United States are
given in Tables 9.5 and 9.6.

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Table 9.5  U.S. Patents (Granted) on Transgenic Plants Developed for Fruit
Skin Color
Patent Named
No. Date Inventors Applicant Area
7678393 March 16, Bradford DB Laboratories Anthocyanin
2010 et al. LLC, Stites, mixture useful
Idaho for topical
and internal
applications
7682637 March 23, Gerald et al. Phenolics LLC, Compositions
2010 Omaha, enriched in
Nebraska total phenols
7727564 June 1, 2010 Kenneth et al. K2A LLC, Fruit-based
Springville, dietary
Utah supplements
7750211 July 6, 2010 Richard et al. Samuel Roberts Production of
Noble flavonoids and
Foundation, isoflavonoids
Ardmore,
Oklahoma
7767416 August 3, Spangenberg Ariculture Flavonoid
2010 et al. Victoria Service biosynthesis in
Pty. Ltd., plants
Altwood,
Australia, and
Agresearch
Ltd., Hamilton,
New Zealand
7855319 December Rommens J.R. Simplot Antioxidant-
21, 2010 et al. Company, containing
Boise, Idaho food
7880059 February 1, Richard et al. The Samuel Proanthocyanins
2011 Roberts Noble to improve
Foundation, forage quality
Ardmore
Oklahoma (US)

9.9  Conclusions and Future Perspectives

The initiation and progression of fruit color is a highly complex phenom-
enon and dependent on several factors, such as fruit development stage,
developmental regulation, phytohormones, and external factors, including

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 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Table 9.6  U.S. Patent Applications on Transgenic Plants Developed for Fruit
Color
Named
Patent No. Date Inventors Applicant Area
20100016565 January 21, Connors Exelixis Plant Anthocyanin
2010 et al. Science, Inc. mutant (ANT1)
in tomato
20100015065 January 21, Matsumoto — Anthocyanin
2010 et al. composition in
food
20100021614 January 28, Nishijima — Bioabsorption of
2010 et al. flavonoid
20100041877 February Tamura — Purified
18, 2010 et al. anthocaynin
20100107277 April 29, Bruguera International Flavonoid
2010 et al. Flower 3,5-hydroxylase
Developments gene
Pty. Ltd.
20100216729 April 30, Gutierrez- — Cancer cell
2010 Uribe growth
et al. inhibition
20100199370 August 5, Levin et al. — Fruits with high
2010 levels of
anthocyanin
and flavonols
20110072539 March 24, Espley — Pigment
2011 et al. production in
plant
—, data not available.

light and temperature. While considering carotenoids and anthocyanins are

major factors in the fruit skin color, a linear set of events can be described.
Fruit skin color variation in fruit is strongly associated with genetic varia-
tion and a combination of bioinformatic aspects (Borevitz et al., 2000; Xie
et al., 2006). Consequently, biotechnological approaches, which include
marker-assisted breeding and genetic transformation, are of particular
importance in fruit skin coloration. This chapter established a clear rela-
tionship between fruit pigmentation and differential transcriptional pat-
terns in the anthocyanin- and carotenoid-related genes during the fruit
color developmental process.
The latest achievements in plant genomics have enabled scientists to
identify the entire array of genes that sustain a plant. The best example is

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publication of the whole genome sequence of grape, providing a valuable tool

and resource to the fruit plant community. This progress in genomic research
has fostered new approaches to address the understanding of physiological
and biochemical pathways. The crux of the technology will be how these prod-
ucts fare against public opinion and the challenges of the current regulatory
systems. However, the functional relationships between this genetic element
and transcriptional activity are still unknown, and more studies are needed
to better characterize the genetic bases of fruit skin coloration. Future expan-
sions in this area of research will depend strongly on solving these problems.

References
Aharoni, A., Ric De Vos, C.H., Wein, M., Sun, Z.K., Greco, R., Kroon, A., Mol,
J.M.N., and Connell, A.P.O. 2001. The strawberry FaMYB1 transcrip-
tion factor suppresses anthocyanin and flavonol accumulation in trans-
genic tobacco. Plant Journal 28, 319–332.
Allan, A.C., Hellens, R.P., and Laing, W.A. 2008. MYB transcription factors
that color our fruit trends. Plant Science 13, 99–102.
Almeida, J.R.M., Amico, E.D., Preuss, A., Carbone, F., de Vos, C.H.R., Deiml,
B., Mourgues, F., Perrotta, G., Fischer, T.C., Bovy, A.G., Martens, S., and
Rosati, C. 2007. Characterization of major enzymes and genes involved
in flavonoid and proanthocyanidin biosynthesis during fruit develop-
ment in strawberry (Fragaria × ananassa). Archives of Biochemistry and
Biophysics 465, 61–71.
Aradhya, M.K., Dangl, G.S., Prins, B.H., Boursiquot, J.M., Walker, M.A.,
Meredith, C.P., and Simon, C.J. 2003. Genetic structure and differentia-
tion in cultivated grape, Vitis vinifera L. Genetic Research 81, 179–192.
Arakawa, O., Shinoda, M., Hiraga, M., and Wang, H. 1999. Comparison of
anthocyanin synthesis of true-to-type ‘Tsugaru’ apple and its red sport
strain. Journal of Horticultural Science and Biotechnology 74, 738–742.
Awad, M.A., de Jager, A., and van Westing, L.M. 2000. Flavonoid and chlo-
rogenic acid level in apple fruit: Characterization of variation. Scientia
Horticulturae 83, 249–263.
Ballinger, W.E., Maness, E.P., Galletta, G.J., and Kushman, L.J. 1972.
Anthocyanin of ripe fruit of a pink-fruited hybrid of highbush blue-
berries, Vaccinium corymbosum L. Journal of the American Society for
Horticulture Science 97, 381–384.
Ban, Y., Honda, C., Hatsuyama, Y., Igarashi, M., Bessho, H., and Moriguchi,
T. 2007. Isolation and functional analysis of a MYB transcription factor
gene that is a key regulator for the development of red coloration in
apple skin. Plant and Cell Physiology 48, 958–970.
Bao, J.S., Cai, Y.Z., Sun, M., Wang, G.Y., and Corke, H. 2005. Anthocyanins
flavonols and free radical scavenging activity of Chinese bayberry
(Myrica rubra) extracts and their color properties and stability. Journal
of Agricultural and Food Chemistry 53, 2327–2332.

288
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Bartley, G.E., and Scolnik, P.A. 1995. Plant carotenoids: pigments for
photoprotection, visual attraction, and human health. Plant Cell 7,
1027–1038.
Ben-Yehudah, G., Korchinsky, R., Redel, G., Ovadya, R., Oren-Shamir, M.,
and Cohen, Y. 2005. Color accumulation patterns and the anthocy-
anin biosynthetic pathway in “Red Delicious” apple variants. Journal of
Horticultural Science and Biotechnology 80, 187–192.
Ben Zvi, M.M., Florence, N.Z., Masci, T., Ovadis, M., Shklarman, E., Ben-
Meir, H., Tzfira, T., Dudareva, N., and Vainstein, A. 2008. Interlinking
showy traits: Co-engineering of scent and color biosynthesis in flowers.
Plant Biotechnology Journal 6, 403–415.
Bhaskarachary, K., Rao, D.S.S., Deosthale, Y.G., and Reddy, V. 1995. Carotene
content of some common and less familiar foods of plant origin. Food
Chemistry 54, 189–193.
Bobe, G., Wang, B., Seeram, N.P., Nair, M.G., and Bourquin, L.D. 2006.
Dietary anthocyanin-rich tart cherry extract inhibits intestinal tumori-
genesis in APC (Min) mice fed suboptimal levels of sulindac. Journal of
Agricultural and Food Chemistry 54, 9322–9328.
Boches, P.S., Peterschmidt, B.C., and Myers, J.R. 2009. Breeding tomato
for increased fruit phenolics. Journal of the American Society for
Horticultural Science 44, 1055–1056.
Bogs, J., Jaffé, F.W., Takos, A.M., Walker, A.R., and Robinson, S.P. 2007. The
grapevine transcription factor VvMYBPA1 regulates proanthocyanidin
synthesis during fruit development. Plant Physiology 143, 1347–1361.
Borevitz, J.O., Xia, Y., Blount, J., Dixon, R.A., and Lamb, C. 2000. Activation
tagging identifies a conserved MYB regulator of phenylpropanoid bio-
synthesis. Plant Cell 12, 2383–2394.
Bornsk, S.M., Ziberna, L., Polak, T., Vanzo, A., Ulrih, N.P., Abram, V., Tramer,
F., and Passamonti, S. 2012. Bilberry and blueberry anthocyanins act as
powerful intracellular antioxidants in mammalian cells. Food Chemistry
134, 1878–1884.
Boss, P.K., Davies, C., and Robinson, S.P. 1996. Anthocyanin composition
and anthocyanin pathway gene expression in grapevine sports differ-
ing in berry skin color. Australian Journal of Grape Wine Research 2,
163–170.
Bovy, A., Schijlen, E., and Hall, R.D. 2007. Metabolic engineering of flavo-
noids in tomato (Solanum lycopersicum): The potential for metabolo-
mics. Metabolomics 3, 399–412.
Bramley, P.M. 2000. Is lycopene beneficial to human health? Phytochemistry
54, 233–236.
Brouillard, R. 1982. Chemical structure of anthocyanins. In Anthocyanins as
Food Colors, Markakis, P. (ed.), 1–40. Academic Press, New York.
Butelli, E., Titta, L., Giorgio, M., Mock, H.P., Matros, A., Peterek, S.,
Schijlen,  E., Hall, R.D., Bovy, A.G., Luo, J., and Martin, C. 2008.
Enrichment of tomato fruit with health-promoting anthocyanin by expres-
sion of select transcription factors. Nature Biotechnology 26, 1301–1308.

289
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Carbone, F., Preuss, A., Vos, R.C.H.D., Amico, E.D., Perrotta, G., Bovy, A.G.,
Martens, S., and Rosati, C. 2009. Developmental, genetic and envi-
ronmental factors affect the expression of flavonoid genes, enzymes
and metabolites in strawberry fruits. Plant and Cell Environment 32,
1117–1131.
Charoensiri, R., Kongkachuichai, R., Suknicom, S., and Sungpuag, P. 2009.
Beta-carotene, lycopene, and alpha-tocopherol contents of selected Thai
fruits. Food Chemistry 113, 202–207.
Chen, K.S., Xu, C.J., Zhang, B., and Ferguson, I.B. 2004. Red bayberry:
Botany and horticulture. Horticultural Reviews 30, 83–114.
Chen, P.N., Chu, S.C., Chiou, H.L., and Kuo, W.H. 2006. Mulberry anthocya-
nins, cyanidin 3-rutinoside and cyanidin 3-glucoside, exhibited an inhib-
itory effect on the migration and invasion of a human lung cancer cell
line. Cancer Letters 235, 248–259.
Corrigan, V.K., Hurst, P.L., and Boulton, G. 1997. Sensory characteristics and
consumer acceptability of ‘Pink Lady’ and other late-season apple culti-
vars. New Zealand Journal of Crop and Horticulture 25, 375–383.
Davies, K.M., Schwinn, K.E., Deroles, S.C., Manson, D.G., Lewis, D.H.,
Bloor, S.J., and Bradley, J.M. 2003. Enhancing anthocyanin production
by altering competition for substrate between flavonol synthase and
dihydroflavonol 4-reductase. Euphytica 131, 259–268.
Deluc, L., Barrieu, F., Marchive, C., Lauvergeat, V., Decendit, A., Richard, T.,
Carde, J.P., Me´rillon, J.M., and Hamdi, S. 2006. Characterization of a
grapevine R2R3-MYB transcription factor that regulates the phenylpro-
panoid pathway. Plant Physiology 140, 499–511.
Deluc, L., Bogs, J., Walker, A.R., Ferrier, T., Decendit, A., Merillon,
J.M., Robinson, S.P., and Barrieu, F. 2008. The transcription factor
VvMYB5b contributes to the regulation of anthocyanin and proantho-
cyanidin biosynthesis in developing grape berries. Plant Physiology
147, 2041–2053.
Dias, M.G., Filomena, M., Camões, G.F.C., and Oliveira, L. 2009. Carotenoids
in traditional Portuguese fruits and vegetables. Food Chemistry 113,
808–815.
Dixon, R.A., and Steele, C.L. 1999. Flavonoids and isoflavonoids—A gold
mine for metabolic engineering. Trends in Plant Science 4, 394–400.
Dunwell, J.M. 2005. Intellectual property aspects of plant transformation.
Plant Biotechnology Journal 3, 371–384.
Einbond, L.S., Reynertson, K.A., Luo, X.D., Basile, M.J., and Kennelly, E.J.
2004. Anthocyanin antioxidants from edible fruits. Food Chemistry 84,
23–28.
Escribano-Bailón, M.T., Santos-Buelga, C., and Rivas-Gonzalo, J.C. 2004.
Anthocyanins in cereals. Journal of Chromatography A 1054, 129–141.
Espley, R.V., Brendolise, C., Chagne, D., Amma, S.K., Green, S., Volz, R.,
Putterill, J., Schouten, H.J., Gardiner, S.E., Hellens, R.P., and Allan, A.C.
2009. Multiple repeats of a promoter segment causes transcription fac-
tor autoregulation in red apples. Plant Cell 21, 168–183.

290
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Fang, Z., Zhang, Y., Lü, Y., Ma, G., Chen, J., Liu, D., and Ye, X. 2009. Phenolic
compounds and antioxidant capacities of bayberry juices. Food
Chemistry 113, 884–888.
Furr, H.C., and Clark, R.M. 1997. Intestinal absorption and tissue distribution
of carotenoids. Journal of Nutritional Biochemistry 8, 364–377.
Gallus, S., Talamini, R., Giacosa, A., Montella, M., Ramazzotti, V., Franceschi,
S., Negri, E., and La Vecchia, C. 2005. Does an apple a day keep the
oncologist away? Annals of Oncology 16, 1841–1844.
Gao, L., and Mazza, G. 1995. Quantitation and distribution of simple and acyl-
ated anthocyanins and other phenolics in blueberries. Journal of Food
Science 59, 1057–1059.
Gil, M.I., Holcroft, D.M., and Kader, A.A. 1997. Changes in strawberry antho-
cyanins  and other polyphenols in response to carbon dioxide treat-
ments. Journal of Agricultural and Food Chemistry 45, 1662–1667.
Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., and Kader, A.A. 2002.
Antioxidant capacities, phenolic compounds, carotenoids, and vitamin C
contents of nectarine, peach, and plum cultivars from California. Journal
of Agricultural and Food Chemistry 50, 4976–4982.
Gonzali, S., Mazzucato, A., and Perata, P. 2009. Purple as a tomato towards
high anthocyanin tomatoes. Trends in Plant Science 14, 237–241.
Granado-Lorencio, F., Olmedilla-Alonso, B., Herrero-Barbudo, C., Blanco-
Navarro, I., Pérez- Sacristán, B., and Blázquez-García, S. 2007. In vitro
bioaccessibility of carotenoids and tocopherols from fruits and vegeta-
bles. Food Chemistry 102, 641–648.
Hannum, S.M. 2004. Potential impact of strawberries on human health: a review
of the science. Critical Reviews of Food Science and Nutrition 44, 1–17.
Harborne, J.B., and Hall, E. 1964. Plant polyphenols-XIII. The systematic
distribution and origin of anthocyanins containing branched trisaccha-
rides. Phytochemistry 3, 453–463.
Holcroft, D.M., and Mitcham, E.J. 1996. Postharvest physiology and handling
of litchi (Litchi chinensis Sonn.). Postharvest Biology and Technology 9,
265–281.
Holden, J.M., Eldridge, A.L., Beecher, G.R., Buzzard, I.M., Bhagwat, A.S.,
Davis, C.S., Douglass, L.W., Gebhardt, E.S., Haytowitz, D., and Schakel,
S. 1999. Carotenoid content of U.S. foods: An update of the database.
Journal of Food Composition and Analysis 12, 169–196.
Holton, T.A., and Cornish, E.C. 1995. Genetics and biochemistry of anthocy-
anin biosynthesis. Plant Cell 7, 1071–1083.
Honda, C., Kotoda, N., Wada, M., Kondo, S., Kobayashi, S., Soejima, J., Zhang,
Z., Tsuda, T., and Moriguchi, T. 2002. Anthocyanin biosynthetic genes
are coordinately expressed during red coloration in apple skin. Plant
Physiology and Biochemistry 40, 955–962.
Hornero-Méndez, D., and Mínguez-Mosquera, M.I. 2000. Xanthophyll esteri-
fication accompanying carotenoid over accumulation in chromoplast of
Capsicum annuum ripening fruits is a constitutive process and useful for
ripeness index. Journal of Agricultural and Food Chemistry 48, 1617–1622.

291
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Hrazdina, G., Parsons, G.F., and Mattick, L.R. 1984. Physiological and bio-
chemical events during development and maturation of grape berries.
American Journal of Enology and Viticulture 35, 220–227.
Huang, P.Y., and Scott, K.J. 1985. Control of rotting and browning of litchi fruit after
harvest at ambient temperatures in China. Tropical Agriculture 62, 2–4.
Hughes, N.M., Morley, C.B., and Smith, W.K. 2007. Coordination of anthocy-
anin decline and photosynthetic maturation in juvenile leaves of three
deciduous tree species. New Phytology 175, 675–685.
Hulshof, P.J.M., Chao, X., Van De Bovenkamp, P., Muhilal, and West, C.E.
1997. Application of a validated method for the determination of provi-
tamin A carotenoids in Indonesia foods of different maturity and origin.
Journal of Agricultural and Food Chemistry 45, 1174–1179.
Jaakola, L., Matta, K., Pirttila, A., Torronen, R., Karenlampi, S., and Hohotola,
A. 2002. Expression of gene involved in anthocyanin biosynthesis in
relation to anthocyanin, proanthocyanin, and flavonol levels during bil-
berry fruit development. Plant Physiology 130, 729–739.
Jaakola, L., Poole, M., Jones, M.O., Karppinen, T.K., Koskimmäki, J.J.,
Hohtola, A., Häggman, H., Fraser, P.D., Manning, K., King, G.J,
Thomson, H., and Seymour, G.B. 2010. A SQUAMOSA MADS box gene
involved in the regulation of anthocyanin accumulation in bilberry fruit.
Plant Physiology 153, 1619–1629.
Jiang, Y.M., Duan, X.W., Joyce, D., Zhang, Z.Q., and Li, L.R. 2004. Advances
in understanding enzymatic browning of harvested litchi fruit. Food
Chemistry 88, 443–446.
Jiang, Y.M., Wang, Y., Song, L.L., Liu, H., Lichter, A., Kerdchoechuen, O.,
Joyce, D.C., and Shi, J. 2006. Production and postharvest characteris-
tics and technology of litchi fruit: An overview. Australian Journal of
Experimental Agriculture 46, 1541–1556.
Jones, C.M., Mes, P., and Myers, J.R. 2003. Characterization and inheritance
of the anthocyanin fruit (Aft) tomato. Journal of Heredity 94, 449–456.
Kayesh, E., Lingfei, S., Nicholas, K.K., Xin, S., Nadira, B., Yanping, Z., Jian,
H., Changnian, S., Zong-Ming, C., and Jinggui, F. 2013. Fruit skin color
and the role of anthocyanin. Acta Physiologiae Plantarum 35, 2879–2890.
Kendrick, R.E., Kerckhoffs, L.H.J., van Tuinen, A., and Koornneef, M. 1997.
Photomorphogenic mutants of tomato. Plant Cell and Environment 20,
746–751.
Kim, S.H., Lee, J.R., Hong, S.T., Yoo, Y.K., and Kim, S.R. 2003. Molecular
cloning and analysis of anthocyanin biosynthesis genes preferentially
expressed in apple skin. Plant Science 165, 403–413.
King, M.C., and Cliff, M.A. 2002. Development of a model for prediction of
consumer liking from visual attributes of new and established apple cul-
tivar. Journal of American Pomological Society 56, 223–229.
Kobayashi, H., Suzuki, S., Tanzawa, F., and Takayanagi, T. 2009. Low expres-
sion of flavonoid 3′,5′-hydroxylase (F3′5′H) associated with cyanidin-
based anthocyanins in grape leaf. American Journal of Enology and
Viticulture 60, 362–367.

292
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Kobayashi, S., Goto-Yamamoto, N., and Hirochika, H. 2004. Retrotransposon-

induced mutations in grape skin color. Science 304, 982.
Kobayashi, S., Goto-Yamamoto, N., and Hirochika, H. 2005. Association
of VvmybA1 gene expression with anthocyanin production in grape
(Vitis vinifera) skin-color mutants. Journal of the Japanese Society for
Horticultural Science 74, 196–203.
Kobayashi, S., Ishimaru, M., Hiraoka, K., and Honda, C. 2002. Myb related
genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin bio-
synthesis. Planta 215, 924–933.
Kong, J.M., Chia, L.S., Goh, N.K., Chia, T.F., and Brouillard, R. 2003. Analysis
and biological activities of anthocyanins. Phytochemistry 64, 923–933.
Kopen, J.M., Happonen, A.M., Mattila, P.H., and Torronen, A.R. 2007.
Contents of anthocyanins and ellagitannins in selected foods con-
sumed in Finland. Journal of Agricultural and Food Chemistry 55,
1612–1619.
Kosar, M., Kafkas, E., Paydas, S., and Baser, K.H.C. 2004. Phenolic composi-
tion of strawberry genotypes at different maturation stages. Journal of
Agricultural and Food Chemistry 52, 1586–1589.
Koshita, Y., Yamane, T., Yakushiji, H., Azuma, A., and Mitani, N. 2011.
Regulation of skin color in ‘Aki Queen’ grapes: Interactive effects of
temperature, girdling, and leaf shading treatments on coloration and
total soluble solids. Scientia Horticulturae 129, 98–101.
Krinsky, N.I., and Johnson, E.J. 2005. Carotenoid actions and their relation to
health and disease. Molecular Aspects of Medicine 26, 459–516.
Kriventsov, V.I., and Arendt, N.K. 1981. Anthocyanins of pomegranate juice.
Trudy Gosuadarstvennogo Nikitskogo, Botanicheskogo Sada 83, 110–116.
Lako, J., Trenerry, V.C., Wahlqvist, M., Wattanapenpaiboon, N., Sotheeswaran,
S., and Premier, R. 2007. Phytochemical flavonols, carotenoids and the
antioxidant properties of a wide selection of Fijian fruit, vegetables and
other readily available foods. Food Chemistry 101, 1727–1741.
Lancaster, J.E. 1992. Regulation of skin color in apples. Critical Review of
Plant Science 10, 487–502.
Lancaster, J.E., Lister, C.E., Reay, P.F., and Triggs, C.M. 1997. Influence of
pigment composition on skin color in a wide range of fruit and veg-
etables. Journal of the American Society for Horticultural Science 122,
594–598.
Lamikanra, O. 1989. Anthocyanins of Vitis rotundifolia hybrid grapes. Food
Chemistry 33, 225–237.
Lee, H.S., and Wicker, L. 1991. Anthocyanin pigments in the skin of lychee
fruit. Journal of Food Science 56, 466–468.
Levy, A., Harel, S., Palevitch, D., Akiri, B., Menagem, E., and Kanner, J. 1995.
Carotenoid pigments and β-carotene in paprika fruits (Capsicum spp.)
with different genotypes. Journal of Agricultural and Food Chemistry 43,
362–366.
Liu, R.H., Liu, J.R., and Chen, B.Q. 2005. Apples prevent mammary tumors in
rats. Journal of Agricultural and Food Chemistry 53, 2341–2343.

293
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lodhi, M.A., and Reisch, B.I. 1995. Nuclear DNA content of Vitis species,
cultivation and other genera of the Vitaceae. Theoretical and Applied
Genetics 90, 11–16.
Lopes-da-Silva, F., Escribano-Bailón, M.T., Perez-Alonso, J.J., Rivas-Gonzalo,
J.C., and Santos-Buelga, C. 2007. Anthocyanin pigments in strawberry.
LWT Food Science and Technology 40, 374–382.
Makus, D.J., and Batlinger, W.E. 1973. Characterization of anthocyanins dur-
ing ripening of fruit of Vaccinium corymbosum L. cv. Wolcott. Journal of
the American Society for Horticulture Science 98, 99–101.
Manning, K. 1998. Isolation of a set of ripening-related genes from straw-
berry, their identification and possible relationship to fruit quality traits.
Planta 205, 622–631.
Marín, A., Ferreres, F., Tomas-Barberan, F.A., and Gil, M.I. 2004.
Characterization and quantitation of antioxidant constituents of sweet
pepper (Capsicum annuum L.). Journal of Agricultural and Food
Chemistry 52, 3861–3869.
Markus, F., Daood, H.G., Kapitany, J., and Biacs, P.A. 1999. Change in the
carotenoid and antioxidant content of spice red pepper (paprika)
as a function of ripening and some technological factors. Journal of
Agricultural and Food Chemistry 47, 100–107.
Martens, S., Preuß, A., and Matern, U. 2010. Multifunctional flavonoid
dioxygenases: Flavonol and anthocyanin biosynthesis in Arabidopsis
­thaliana L. Phytochemistry 71, 1040–1049.
Martinez, M.V., and Whitaker, J.R. 1995. The biochemistry and control of
enzymatic browning. Trends in Food Science and Technology 6, 195–200.
Mathews, H., Clendennen, S.K., Caldwell, C.G., Liu, X.L., Connors, K.,
Matheis, N., Schuster, D.K., Menasco, D.J., Wagoner, W., Lightner, J.,
and Wagner, R. 2003. Activation tagging in tomato identifies a transcrip-
tional regulator of anthocyanin biosynthesis, modification, and trans-
port. Plant Cell 15, 1689–1703.
Mazza, G., and Miniati, E. 1993. Anthocyanins in Fruits, Vegetables and Grains.
CRC Press, Taylor and Francis, Boca Raton, FL.
Mazza, G., and Velioglu, Y.S. 1992. Anthocyanin and other phenolic com-
pounds in fruits of red-flesh apples. Food Chemistry 43, 113–117.
Mes, P.J., Boches, P., Myers, J.R., and Durst, R. 2008. Characterization of
tomatoes expressing anthocyanin in the fruit. Journal of the American
Society for Horticultural Science 133, 262–269.
Minguez-Mosquera, M.I., and Hornero-Mendez, D. 1994. Changes in carot-
enoid esterification during the fruit ripening of Capsicum annuum cv.
Bola. Journal of Agricultural and Food Chemistry 42, 640–644.
Minguez-Mosquera, M.I., Gandul-Rojas, B., Garrido-Fernandez, J., and
Gallardo-Guerrero, L. 1990. Pigments present in virgin olive oil. Journal
of American Oil Chemists Society 67, 192–196.
Mori, K., Sugaya, S., and Gemma, H. 2005. Decreased anthocyanin biosyn-
thesis in grape berries grown under elevated night temperature condi-
tion. Scientia Horticulturae 105, 319–330.

294
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Muir, S.R., Collins, G.J., Robinson, S., Hughes, S., Bovy, A., De Vos, C.H.R.,
van Tuinen, A.J., and Verhoeyen, M.E. 2001. Overexpression of petunia
chalcone isomerase in tomato results in fruit containing increased lev-
els of flavonols. Nature Biotechnology 19, 470–474.
Niu, S.S., Xu, C.J., Zhang, W.S., Zhang, B., Li, X., Wang, K.L., Ferguson, I.B.,
Allan, A.C., and Chen, K.S. 2010. Coordinated regulation of anthocyanin
biosynthesis in Chinese bayberry (Myrica rubra) fruit by a R2R3 MYB
transcription factor. Planta 231, 887–899.
Oomah, B.D., and Mazza, G. 1999. Health benefits of phytochemicals from
selected Canadian crops. Trends in Food Science and Technology 10, 193–198.
Oren-Shamir, M. 2009. Does anthocyanin degradation play a significant role
in determining pigment concentration in plants? Plant Science 177,
310–316.
Palapol, Y., Ketsa, S., Wang, K.L., Ferguson, B.I., and Allan, A.C. 2009. A MYB
transcription factor regulates anthocyanin biosynthesis in mangosteen
(Garcinia mangostana L.) fruit during ripening. Planta 229, 1323–1334.
Pantelidis, G.E., Vasilakakis, M., Manganaris, G.A., and Diamantidis, G. 2007.
Antioxidant capacity, phenol, anthocyanin and ascorbic acid contents
in raspberries, blackberries, red currants, gooseberries and Cornelian
cherries. Food Chemistry 102, 777–783.
Pecket, R.C., and Small, C.J. 1980. Occurrence, location and development of
anthocyanoplasts. Phytochemistry 19, 2571–2576.
Pierantoni, L., Dondini, L., De Franceschi, P., Musacch, S., Winkel, B.S.J.,
and Sansavini, S. 2010. Mapping of an anthocyanin regulating MYB
transcription factor and its expression in red and green pear (Pyrus
­communis). Plant Physiology and Biochemistry 48, 1020–1026.
Pomar, F., Novo, M., and Masa, A. 2005. Varietal differences among the antho-
cyanin profiles of 50 red tablegrape cultivars studied by high perfor-
mance liquid chromatography. Journal of Chromatography 109, 34–41.
Povero, G., Gonzali, S., Bassolino, L., Mazzucato, A., and Perata, P. 2011.
Transcriptional analysis in high-anthocyanin tomatoes reveals ­synergistic
effect of Aft and atv genes. Journal of Plant Physiology 168, 270–279.
Prasad, U.S., and Jha, O.P. 1978. Changes in pigmentation patterns during
litchi ripening: Flavonoid. Plant Biochemistry Journal 5, 44–49.
Pray, C.E., and Naseem, A. 2007. Supplying crop biotechnology to the poor:
Opportunities and constraints. Journal of Developmental Studies 43, 192–217.
Quattrocchio, F., Wing, J.F., Van der Woude, K., Mol, J.N.M., and Koes, R.
1998. Analysis of bHLH and MYB domain proteins: Species-specific reg-
ulatory differences are caused by divergent evolution of target anthocy-
anin genes. Plant Journal 13, 475–488.
Rao, A.V., and Rao, L.G. 2007. Carotenoids and human health. Pharmacology
Research 55, 207–216.
Rasmussen, S.E., Frederiksen, H., Krogholm, K.S., and Poulsen, L. 2005.
Dietary proanthocyanidins: occurrence, dietary intake, bioavailability,
and protection against cardiovascular disease. Molecular Nutrition and
Food Research 49, 159–174.

295
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Rice-Evans, C., Miller, N.J., and Paganga, G. 1997. Antioxidant properties of

phenolic compounds. Trends in Plant Science 2, 152–159.
Rick, C.M., Reeves, A.F., and Zobel, R.W. 1968. Inheritance and linkage ­relations
of four new mutants. Tomato Genetics Cooperative Report 18, 34–35.
Rodriguez-Amaya, D.B. 2001. A Guide to Carotenoid Analysis in Foods.
International Life Sciences Institute, ILSI Press, Washington, DC.
Rodriguez-Amaya, D.B., and Kimura, M. 2004. HarvestPlus Handbook for
Carotenoid Analysis. HarvestPlus Technical Monograph, Series 2.
International Food Policy Research Institute and International Center
for Tropical Agriculture, Washington, DC.
Salvatierra, A., Pimentel, P., Moya-Leon, M.A., Caligari, P.D.S., and Herrera,
R. 2010. Comparison of transcriptional profiles of flavonoid genes
and anthocyanin contents during fruit development of two botanical
forms of Fragaria chiloensis ssp. chiloensis. Phytochemistry 71, 1839–1847.
Sapir, M., Oren-Shamir, M., Ovadia, R., Reuveni, M., Evenor, D., Tadmor, Y.
et al., 2008. Molecular aspects of anthocyanin fruit tomato  in relation
to high pigment-I. Journal of Heredity 99, 292–303.
Sarni-Manchado, P., Roux, E.L., Guerneve, C.L., Lozano, Y., and Cheynier, V.
2000. Phenolic composition of litchi fruit pericarp. Journal of Agricultural
and Food Chemistry 48, 5995–6002.
Schirrmacher, G., and Schempp, H. 2003. Antioxidative potential of flavonoid-
rich extracts as new quality marker for different apple varieties. Journal
of Applied Botany 77, 163–166.
Scott, K.J., Thurnham, D.I., Hart, D.J., Bingham, S.A., and Day, K. 1996. The
correlation between the intake of lutein, lycopene and β-carotene from
vegetables and fruits, and blood plasma concentrations in a group of
women aged 50–65 years in the UK. British Journal of Nutrition 75,
409–418.
Setiawan, B., Sulaeman, A., Giraud, D.W., and Driskell, J.A. 2001. Carotenoid
content of selected Indonesian fruits. Journal of Food Composition and
Analysis 14, 169–176.
Simpson, K.L. 1985. Chemical changes in natural food pigments. In Chemical
Changes in Food during Processing, Richardson, T., and Finley, J.W.
(eds.). Van Nostrand Reinhold Company, New York, 409–437.
Sparvoli, F., Martin, C., Scinza, A., Gavazzi, G., and Tonelli, C. 1994. Cloning and
molecular analysis of structural genes involved in flavonoid and stilbene bio-
synthesis in grape (Vitis vinifera L.). Plant Molecular Biology 24, 743–755.
Sugawara, T., and Igarashi, K. 2008. Cultivar variation and anthocyanins
and rutin content in sweet cherries (Prunus avium L.). Journal of the
Japanese Society for Food Science and Technology—Nippon Shokuhin
Kagaku Kogaku Kaishi 55, 239–244.
Takos, A.M., Jaffe, F.W., Jacob, S.R., Bogs, J., Robinson, S.P., and Walker, A.R.
2006. Light-induced expression of a MYB gene regulates anthocyanin
biosynthesis in red apple. Plant Physiology 142, 1216–1232.

296
 ROLE O F P IGMENTS D URING F RUIT RI P ENING

Takyi, E.E.K. 2001. Bioavailability of carotenoids from vegetables versus sup-

plements. In Vegetables, Fruits, and Herbs in Health Promotion, Watson,
R.R. (ed.). CRC Press, Boca Raton, Fl, 19–31.
Tanaka, Y., and Ohmiya, A. 2008. Seeing is believing: engineering antho-
cyanin and carotenoid biosynthetic pathways. Current Opinion in
Biotechnology 19, 190–197.
Tee, E.-S., and Lim, C.-L. 1991. Carotenoid composition and content of
Malaysian vegetables and fruits by the AOAC and HPLC methods. Food
Chemistry 41, 309–339.
Torre, L.C., and Barritt, B.H. 1977. Quantitative evaluation of Rubus fruit
anthocyanin pigments. Journal of Food Science 42, 488–490.
Torres, C.A., Davies, N.M., Yanez, J.A., and Andrews, P.K. 2005. Disposition
of selected flavonoids in fruit tissues of various tomato (Lycopersicon
esculentum Mill.) genotypes. Journal of Agricultural and Food Chemistry
53, 9536–9543.
Torres, D.C., Díaz-Maroto, M.C., Hermosín-Gutiérrez, I., and Pérez-Coello,
M.S. 2010. Effect of freeze-drying and oven-drying on volatiles and
­phenolics composition of grape skin. Analytica Chimica Acta 660, 177–182.
Tulipani, S., Mezzetti, B., Capocasa, F., Bompadre, S., Beekwilder, J., de Vos,
C.H.R., Capanoglu, E., Bovy, A., and Battino, M. 2008. Antioxidants, phe-
nolic compounds, and nutritional quality of different strawberry geno-
types. Journal of Agricultural and Food Chemistry 56, 696–704.
Tunen, V.A.J., and Mol, J.N.M. 1991. Control of flavonoids synthesis and
manipulation of flower color. Plant Biotechnology 2, 94–125.
Ubi, B.E., Honda, C., Bessho, H., Kondo, S., Wada, M., Kobayashi, S., and
Moriguchi, T. 2006. Expression analysis of anthocyanin biosynthetic
genes in apple skin. Effect of UV-B and temperature. Plant Science 170,
571–578.
Valcheva-Kuzmanova, S.V., and Belcheva, A. 2007. Colon-available raspberry
polyphenols exhibit anti-cancer effects on in vitro models of colon
­cancer. Journal of Carcinogenesis 6, 4.
Wada, L., and Ou, B.X. 2002. Antioxidant activity and phenolic content of
Oregon caneberries. Journal of Agricultural and Food Chemistry 50,
3495–3500.
Walker, A.R., Lee, E., Bogs, J., McDavid, D.A.J., Thomas, M.R., and Robinson,
S.P. 2007. White grapes arose through the mutation of two similar and
adjacent regulatory genes. Plant Journal 49, 772–785.
Wang, K.L., Bolitho, K., Grafton, K., Kortstee, A., Karunairetnam, S., McGhie,
T.K., Espley, R.V., Hellens, R.P., and Allan, A.C. 2010. An R2R3 MYB
transcription factor associated with regulation of the anthocyanin bio-
synthetic pathway in Rosaceae. BMC Plant Biology 10:50.
Wang, S.Y., and Lin, H.S. 2000. Antioxidant activity in fruit and leaves of black-
berry, raspberry and strawberry varies with cultivar and developmental
stage. Journal of Agricultural and Food Chemistry 48, 140–146.

297
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Wieruszewski, J.B. 2002. Astaxanthin bioavailabity, retention efficiency and

kinetics in Atlantic salmon (Salmo salar) as influenced by pigment con-
centration and method of administration (kinetics only). Master thesis,
Simon Fraser University, Ottawa, Canada.
Winkel-Shirley, B. 2001. Flavonoid biosynthesis: A colorful model for genet-
ics, biochemistry, cell biology, and biotechnology. Plant Physiology 126,
485–493.
Wu, X., Beecher, G.R., Holden, J.M., Haytowitz, D.B., Gebhardt, S.E., and
Prior, R.L. 2006. Concentrations of anthocyanins in common foods in
the United States and estimation of normal consumption. Journal of
Agricultural and Food Chemistry 54, 4069–4075.
Xie, D.Y., Sharma, S.B., Wright, E., Wang, Z.Y., and Dixon, R.A. 2006.
Metabolic engineering of proanthocyanidins through co-expression of
anthocyanin reductase and the PAP1MYB transcription factor. Plant
Journal 45, 895–907.
Yahia, E.M., Ornelas-Paz, J.J., and Gardea, A. 2006. Extraction, separation
and partial identification of ‘Ataulfo’ mango fruit carotenoids. Acta
Horticulturae 712, 333–338.
Yano, M., Kato, M., Ikoma, Y., Kawasaki, A., Fukazawa, Y., Sugiura, M.,
Matsumoto, H., Oohara, Y., Nagao, A., and Ogawa, K. 2005. Quantitation
of carotenoids in raw and processed fruits in Japan. Food Science and
Technology Research 11, 13–18.
Zhang, D.L., Quantick, P.C., and Grigor, J.M. 2000. Changes in phenolic com-
pounds in litchi (Litchi chinensis Soon.) fruit during postharvest stor-
age. Postharvest Biology and Technology 19, 165–172.
Zhang, S.M., Gao, Z.S., and Xu, C.J., Chen, K., Wang, G., Zheng, J., and Lu,
T. 2009. Genetic diversity of Chinese bayberry (Myrica rubra Sieb.
et Zucc.) accessions revealed by amplified fragment length polymor-
phism. HortScience 44, 487–491.
Zhang, W.S., Li, X., Zheng, J.T., Wang, G.Y., Sun, C.D., Ferguson, I.B., and
Chen, K.S. 2008. Bioactive components and antioxidant capacity of
Chinese bayberry (Myrica rubra Sieb. and Zucc.) fruit in relation to
fruit maturity and postharvest storage. European Food Research and
Technology 227, 1091–1097.
Zhang, Z.Q., Pang, X.Q., Yang, C., Ji, Z.L., and Jiang, Y.M. 2004. Purification
and structural analysis of anthocyanins from litchi pericarp. Food
Chemistry 84, 601–604.

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Chapter 10

Molecular Regulation
of Fruit Ripening
Ajay Arora
Indian Agricultural Research Institute, New Delhi, India

Abstract 300
10.1 Introduction 300
10.2  Transcriptional Control of Fruit Ripening 301
10.3  Hormonal and Transcriptional Regulation during
Ripening 305
10.3.1 Ethylene 306
10.3.2 Auxins 308
10.3.3 Gibberellins 308
10.3.4  Abscisic Acid 308
10.4  Epigenetic Regulation of Fruit Development
and Ripening 309
10.5  Ripening Pathways and Associated Fruit Quality 312
10.5.1  Sugar Accumulation 313
10.5.2  Cell Walls and Fruit Shelf Life 313
10.5.3  Color, Flavor, and Nutrition in Fruits 314
10.5.3.1 Chloroplast-to-Chromoplast
Conversion 314
10.5.3.2  Flavor and Aroma Production 315
10.5.3.3  Control of Color and Texture
Changes 316

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10.6 Human Nutrition and Fruits 317

10.6.1 Human Nutrition: Functional Genomics/
Systems Approach 320
10.7 Horticultural Crop Improvement 321
10.8 Genetic Manipulation of Ripening Regulatory Genes 322
10.9 Conclusions and Future Perspectives 322
References 324

Abstract
Fruit ripening is a highly coordinated developmental process that coincides
with seed maturation. Regulated expression of thousands of genes controls
fruit softening as well as accumulation of pigments, sugars, acids, and
volatile compounds that increase attraction to animals. A combination of
molecular tools and ripening-affected mutants has permitted researchers
to establish a framework for the control of ripening. Key to crop improve-
ment is a deeper understanding of the processes underlying fruit ripening.
In tomato, mutations blocking the transition to ripe fruits have provided
insights into the role of ethylene and its associated molecular networks
involved in the control of ripening. However, the role of other plant hor-
mones is still poorly understood. Translation of information from tomato
to other fleshy-fruited species indicates that regulatory networks are con-
served across a wide spectrum of angiosperm fruit morphologies. In this
chapter, we describe how plant hormones, transcription factors, and epigen-
etic changes are intimately related to provide tight control of the ripening
process. These discoveries are likely to have a major impact on strategies
for crop improvement through genetic manipulation of ripening regulatory
genes in fruit-bearing species. The findings from comparative genomics
and system biology approaches are also discussed. Recent developments in
the sequencing of angiosperm genomes have provided the foundation for a
step change in crop improvement through the understanding and harness-
ing of genome-wide genetic and epigenetic variation.

10.1 Introduction
Ripening represents a summation of physiological and biochemical pro-
cesses of fleshy fruits, including, but not limited to, degreening and accu-
mulation of colored pigments for attraction, textural changes associated
with cell wall metabolism and cell turgor variation leading to softening,
and metabolic changes related to flavor and nutrient composition, generally
associated with accumulation of sugars, acids, and volatiles culminating

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in a diverse array of tastes and smells, varying among species (Klee and
Giovannoni, 2011). In climacteric fruit such as apple, tomato, banana, and
most stone fruits, the onset of ripening coincides with a burst of respira-
tion and ethylene, and ethylene is necessary to confer these physiological
changes. However, elevated respiration and ethylene are not required for
normal ripening in fruit classified as nonclimacteric, including important
fruit crops such as grape, citrus, and strawberry, indicating ethylene-­
independent ripening events are important in many species, including
those undergoing climacteric ripening. Investigations into the molecular
and genetic regulation of ripening have focused largely on the model sys-
tem of tomato due to its dramatic ripening process, short life cycle, ame-
nability to genetic transformation, and the plethora of available molecular
and genetic resources, including its recently sequenced genome (Tomato
Genome Consortium, 2012). The agricultural importance and unique attri-
butes of many additional species have resulted in the characterization of
ripening physiology, metabolism, and molecular biology, which is high-
lighted in this chapter as well. The recent history of ripening molecular
biology can be divided into several distinct, though sometimes overlapping,
areas, including the elucidation of cell wall–modifying proteins and their
roles in textural modification, the characterization of ethylene synthesis
and signaling, the genes and pathways contributing to pigment accumula-
tion, the regulatory process associated with fruit plastid development, the
biochemistry of sensory attributes, and the overall genetic regulation of the
ripening process. We will attempt to highlight all areas, with emphasis on
primary regulators that are likely to be functional and conserved among
many fleshy fruit–­producing species.

10.2  Transcriptional Control of Fruit Ripening

Fruits are a distinctive characteristic of angiosperms. They occur today in
a wide variety of forms and types. The ancestral fruit, dry and dehiscent,
probably emerged in the early Cretaceous period; fleshy fruits appeared
later in the Cretaceous or early Tertiary (Eriksson et al., 2000). The diver-
sification of fruits from a dry dehiscent form to a fleshy drupe or berry
correlated with the rise of vertebrates, the main agents of seed dispersal
(Knapp, 2002). The maturation of fruits is a complex and highly coordinated
developmental process. In fleshy fruits, ripening results in the production of
a succulent, flavorful, and soft pericarp that attracts animals and facilitates
seed dispersal (Giovannoni, 2001). In addition to softening, fruits normally
exhibit increased accumulation of sugars, acids, pigments, and volatiles
that increase interest and palatability to animals. Moreover, fruits are an
important source of supplementary diet, providing minerals, ­ v itamins,
fibers, and antioxidants for humans. From an agronomical point of view,

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nutritional value, flavor, processing qualities, and shelf life ­determine the
quality of fruits.
The main changes associated with ripening include color (loss of
green color and increase in nonphotosynthetic pigments that vary depend-
ing on species and cultivar), firmness (softening by cell wall–degrading
activities and alterations in cuticle properties), taste (increase in sugar
and decline in organic acids), and flavor (production of volatile compounds
­providing the characteristic aroma).
The comprehensive molecular analyses of ethylene synthesis and sig-
nal transduction during ripening suggest the presence of additional ­regulatory
systems that coordinate the ethylene burst and production of this necessary
ripening hormone. A number of interesting tomato ripening mutants have
been collected by breeders and geneticists, and their subsequent physiologi-
cal characterization suggests that they may represent defects in ripening
regulatory systems. These mutations include the r­ipening-inhibitor (rin),
non-ripening (nor), and colorless non-ripening (Cnr) mutations that are non-
allelic but share common physiological characteristics in that all (1) develop
to the mature green stage, where the fruit is full size and the seeds are
mature, yet do not advance to ripening; (2) fail to undergo the climacteric
rise in respiration or produce ripening-associated ethylene; (3) do not ripen
in response to exogenous ethylene; and yet (4) respond to ethylene in other
tissues and fruit in which ethylene-responsive genes are induced (Eriksson
et al., 2004; Manning et al., 2006; Giovannoni, 2007).
Together, these characteristics suggest that all three mutations
impact central ripening phenomena necessary for the induction of ethyl-
ene, in addition to activities for which ethylene alone cannot compensate.
In this regard, it is a reasonable hypothesis that such genes might represent
conserved regulators impacting ripening even in nonclimacteric fruits.
All three genes have been isolated by positional cloning, and all encode
transcription factors. There is only one described rin mutation, and it also
displays a large sepal, or macrocalyx, phenotype. The rin locus encodes a
MADS-box transcription factor termed RIN-MADS, which is induced at the
onset of ripening (Vrebalov et al., 2002). MADS-box genes are developmen-
tal regulators conserved in eukaryotes and associated with floral develop-
ment in plants, including members of the ABC model of floral development
(Coen and Meyerowitz, 1991). RIN-MADS is a member of the SEPALLATA
clade associated in Arabidopsis with E function activities that determine the
growth and development of floral organs (Zahn et al., 2005).
Phylogenetic analysis of the RIN gene and examination of the tomato
genome sequence (http://solgenomics.net) indicate two additional genes
compared to Arabidopsis, including RIN-MADS, with the additional gene
having no transcriptional support in the public tomato expressed sequence
tag (EST) collection. This suggests that it may have very limited expres-
sion or be a pseudogene. The RIN-MADS gene resides on c­ hromosome

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5  adjacent to the tomato APETALA1 ortholog, MACROCALYX (MC).

The rin mutation results from a spontaneous deletion that removes the
C-terminus of the RIN-MADS gene and approximately 1 kb of the sequence
separating RIN-MADS from MC. The result is expression of a chimeric
RIN-MADS/MC mRNA that has no apparent RIN-MADS or MC function.
The ripening and large sepal phenotypes of the rin allele were recreated
independently by antisense repression of RIN-MADS and MC, respectively
(Vrebalov et al., 2002). Screening of a strawberry fruit cDNA library with a
tomato RIN-MADS cDNA yielded a homologous gene whose expression was
elevated during ripening. The identification of a strawberry homolog of RIN-
MADS indicates that transcriptional control of ripening is conserved among
climacteric and nonclimacteric species (Vrebalov et al., 2002; Seymour et al.,
2011). Similar banana genes associated with both peel and pulp ripening have
also been described, although they remain to be functionally tested (Elitzur
et al., 2010). If these genes indeed control ripening, this would suggest that
regulation of ripening is a conserved function of the MADS-box family
preceding the divergence of dicots and monocots. It is noteworthy that the
original rin allele is extensively used in tomato hybrid seed production for
heterozygous varieties characterized by long shelf life and fruit firmness,
demonstrating through its practical utility that RIN-MADS exerts broad con-
trol over the ripening process (color, flavor, and texture).
RIN-MADS, as with many genes of this family, has been shown to
interact with CArG-box elements in promoters of ACS2 and ACS4, in addi-
tion to cell wall hydrolases, indicating that it interacts with promoters of
genes representing a range of ripening activities (Fujisawa et al., 2011).
CNR encodes a transcription factor of the SQUAMOSA PROMOTER
BINDING PROTEIN (SPBP) family, which is regulated at least in part by
changes in promoter methylation, in conjunction with ripening, that do not
occur in the Cnr mutant (Manning et al., 2006). CNR expression is reduced
in the rin mutant, suggesting that it may act downstream of RIN-MADS in
the regulatory hierarchy, although the relationship among these regulators
and ethylene is likely not linear. SPBP targets include MADS-box genes,
and the tomato ortholog of the Arabidopsis FRUITFUL (FUL) MADS-box
gene, TDR4, is substantially reduced in Cnr, suggesting that it may be a
CNR target and a regulator through which CNR exerts its effects (Manning
et al., 2006). Although TDR4 repression was reported to yield no obvious
ripening phenotype in tomato, antisense of an orthologous gene in bilberry
resulted in reduced fruit pigmentation (Jaakola et al., 2010). Most of the
well-characterized tomato ripening mutants have been cloned, but addi-
tional regulators certainly remain to be identified.
The transcriptional regulators described above represent positive
effectors of the ripening pathways in that their absence through mutation
or transgenic intervention results in reduced ripening phenotypes. A tomato
APETALA2 (AP2) gene family member, SlAP2a has recently been shown to

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be a negative regulatory of ripening (Chung et al., 2010; Karlova et al., 2011).

SlAP2a is a member of the AP2 subfamily of the AP2/ERF super family, as
it harbors two rather than one conserved AP2 domain. SlAP2a is induced
­during ripening, although its repression resulted in accelerated ripening, ele-
vated ethylene production, and altered carotenoid accumulation. All of these
phenotypes were specific to the fruit, suggesting SlAP2a is a fruit-­specific
and ripening-related suppressor of ethylene production and ripening.
It is clear that multiple transcription factors influence the ripening
process and that gene family expansion driven by polyplodization events
contributed to the reservoir of genes available for the present set of ripening
regulators. The apparent conservation of RIN-like MADS-box genes in rip-
ening control prior to the evolutionary split of monocots and dicots suggests

Transcription factors
Epigenetic dynamics

Ethylene biosynthesis and signaling

Ripening Carotenoid biosynthesis

Antioxidant production
Fruit softening
Flavour and aroma
Colour and texture
Chloroplast to chromoplast

Senescence

Figure 10.1  Molecular ripening components in fruits. Ripening regulation is

governed by transcription factors and epigenome dynamics, which further
impact ethylene synthesis and signal transduction that influence ripening-
associated processes and pathways contributing to fruit quality.

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a conserved ripening function in this important family of floral development

regulators. The contrast in activities of AGAMOUS clade MADS-box genes
suggests other members of this family have diverged in terms of specific
function while retaining their roles in seed dispersal via distinct activities in
very different (dry vs. fleshy) fruit types. The fact RIN-MADS, NOR, CNR,
and TAGL1 all play necessary roles in ripening indicates that they are all
involved either together or separately in ripening control prior to ethylene.
The fact that ectopic expression of TAGL1 can apparently complement the
rin and nor mutations, combined with its earlier expression, suggests a more
primary position in this regulatory network (Giménez et al., 2010), although
it is curious that the rin and nor mutants retain normal TAGL1 expression
without ripening (Vrebalov et al., 2009), suggesting that the full complexity
of this regulatory mechanism remains to be uncovered. The primary role of
these regulators and their impact on ethylene- and non-ethylene-mediated
ripening activities are summarized in Figure 10.1.

10.3  Hormonal and Transcriptional

Regulation during Ripening
A number of important advances in our understanding of mechanisms that
regulate ripening have also come from the characterization of monogenic
tomato mutants, including ripening-inhibitor (rin), non-ripening (nor), color-
less non-ripening (Cnr), green-ripe (Gr), green-flesh (gf ), high pigment 1 (hp1),
high pigment 2 (hp2), and never-ripe (Nr) (Lanahan et al., 1994; Mustilli
et al., 1999; Vrebalov et al., 2002; Liu et al., 2004; Barry and Giovannoni, 2006;
Manning et al., 2006; Barry et al., 2008). The rin mutant encodes a partially
deleted MADS-box protein of the SEPALLATA clade (SEP4) (Hileman
et al., 2006), whereas Cnr is an epigenetic change that alters the promoter
methylation of the SQUAMOSA promoter binding (SPB) protein. NOR is a
member of the NAC-domain transcription factor family (Giovannoni, 2007).
A recent study in which the transcriptome, proteome, and targeted metabo-
lite analysis were combined during development and ripening of nor and rin
mutants has helped to refine the ethylene-regulated expression of down-
stream genes and added to our knowledge the role of this hormone in both
protein and metabolite regulation in tomato ripening (Osorio et al., 2011).
These data supported the view that nor and rin act together in a cascade to
control ripening (Giovannoni et al., 1995; Thompson et al., 1999) and also
suggested that nor has a more global effect on ethylene/ripening-related
gene expression than rin, which indicates that nor likely operates upstream
of rin. Recently, using a combined approach based on chromatin immuno-
precipitation and transcriptome analysis, Fujisawa et al. (2013) provided
evidence that RIN interacts with the promoters of more than 200 genes,

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modulating the expression of its targets by activation or repression. RIN

target genes are major regulators of ripening control, such as CNR and
NOR, or belong (Martel et al., 2011) to well-known pathways active during
the transition from green to ripe fruits (e.g., carotenoid accumulation, chlo-
rophyll breakdown, ethylene synthesis, and perception).

10.3.1 Ethylene
The role of ethylene in fruit ripening has been reviewed many times and
is therefore covered only briefly here. Fleshy fruits have historically been
divided into two classes: climacteric (e.g., tomato, apples, and banana)
and nonclimacteric (e.g., grape, strawberry, and citrus). Climacteric fruits
show a burst of respiration at the onset of ripening, along with a large rise
in ethylene production. Ripening in climacteric fruits can also be initiated
by exposure to exogenous ethylene. In nonclimacteric fruits, the respira-
tory increase is absent and ethylene does not appear to be critical for the
ripening process. Direct evidence that ethylene is critical for the induc-
tion of ripening in climacteric fruit came from work with tomato, where
antisense genes were used to suppress the expression of ACO1 and ACS2,
which encode ACC oxidase (ACO) and ACC synthase (ACS), respectively,
the major enzymes involved in ethylene biosynthesis (Grierson, 2013).
Autocatalytic ethylene biosynthesis (system 2 ethylene biosynthesis)
(Klee and Giovannoni, 2011) is operational during ripening; this involves
new forms of ACO and ACS that produce much higher levels of ethylene and
are not subject to autoinhibition. Ethylene is then perceived by a family of
copper binding membrane-associated receptors, several of which—tomato
ETHYLENE RESPONSE4 (LeETR4), LeETR6, and NR (which results in
the phenotype of the Never-ripe [Nr] tomato mutant)—show significantly
increased expression at the onset of ripening (Klee and Giovannoni, 2011).
The receptors interact with a Raf kinase–like protein, constitutive triple
response1 (CTR1), and repress ethylene responses. When the receptors
bind ­ethylene, this inhibition is relieved and a signaling process leads to
the ethylene response (Grierson, 2013; Klee and Giovannoni, 2011). This
involves activation of the ETHYLENE INSENSITIVE3 (EIN3) and EIN3-
like (EIL) transcription factors by EIN2. In the nonclimacteric pepper fruit,
EIL-like genes are induced during ripening (Lee et al., 2012), suggesting
common systems for downstream signal transduction in climacteric and
nonclimacteric fruits, which in the latter occurs in the absence of ethylene
biosynthesis. In climacteric fruit, at least, ethylene production enhances
the expression of some regulatory and structural genes. Ethylene does
not act to regulate ripening in isolation, but rather acts in concert with
other plant hormones. Lin et al. (2008) identified a tetratricopeptide (TPR)

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repeat protein, SlTPR1, from tomato that interacts with the ethylene recep-
tors LeETR1 and NR. Overexpression of SlTPR1 in tomato enhances
ethylene responses while causing reduced expression of the early auxin-
responsive gene IAA9.
In strawberry, which has emerged as a prime model of nonclimac-
teric fruit ripening, ethylene is relatively high in green fruits, decreases
in white fruits, and finally increases again at the red stage of ripening
(Perkins-Veazie et al., 1996; Iannetta et al., 2006). Interestingly, this last
increase is accompanied by an enhanced respiration rate that resembles
the one that occurs in climacteric fruits at the onset of ripening (Iannetta
et al., 2006). For better understanding of the function of ethylene during
strawberry ripening, different approaches have been used. External appli-
cation of ethylene caused the downregulation of several cell wall–related
genes, such as β-galactosidase, pectin methylesterase, or β-xylosidase
(Trainotti et al., 2001; Castillejo et al., 2004; Bustamante et al., 2009),
while the expression of other genes, such as the expansin FaEXP2 (Civello
et  al., 1999), was ethylene insensitive. Recent studies at transcriptomic
and metabolomic levels in transgenic strawberry fruits with decreased
ethylene sensitivity indicate that ethylene action is required for normal
fruit development, acting differently in the two parts of strawberry fruit,
the achenes and receptacle. These results show that, although not as rel-
evant as in climacteric fruits, ethylene may nevertheless play a role in
strawberry fruit ripening.
Recent comparative transcriptome and metabolome studies dur-
ing the maturation processes of climacteric and nonclimacteric fruits
(tomato and pepper, respectively) suggest that both species have similar
ethylene-mediated signaling components. In pepper, the regulation of
­
these genes is, however, clearly different and may reflect altered ethylene
sensitivity or regulators other than ethylene in tomato (Osorio et al., 2012).
Unlike the situation described in tomato, the ethylene biosynthesis genes,
­a minocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase,
are not induced in pepper. However, genes downstream of ethylene percep-
tion, such as cell wall–related genes, ethylene response factor 3 (ERF3),
and carotenoid biosynthesis genes, are upregulated during pepper fruit
ripening (Osorio et al., 2012). Other commonly regulated genes between
climacteric and nonclimacteric fruits have been described. In strawberry, a
SEPALLATA gene (SEP1/2; MADS-box) is needed for normal development
and ripening (Seymour et al., 2011). Similarly, in banana, which is classified
as a climacteric fruit, the MADS-box SEP3 gene also displays ripening-
related expression (Elitzur et al., 2010). In apple, MADS2 gene expression is
also associated with fruit firmness (Cevik et al., 2010), whereas in bilberry
fruit, the SQUAMOSA MADS-box ortholog of the TDR4 gene in tomato has
a role in regulation of anthocyanin biosynthesis (Jaakola et al., 2010).

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10.3.2 Auxins
Current knowledge about the role of hormones—other than ethylene—in
the development and ripening of climacteric and nonclimacteric fruits is
limited. In tomato, pepper, banana, muskmelon, and strawberry, the most
abundant free auxin, indole-3-acetic acid (IAA), has been reported to
decline prior to the onset of ripening; this reduction was accompanied by
an increase of its conjugated form (IAA-Asp) (Bottcher et al., 2010). The
conjugation reaction is catalyzed by the IAA-amino synthase gene (GH3).
In tomato, 15 members of the GH3 gene family have been described, but
only for 2 of them is the pattern of expression associated with ripening
(Kumar et al., 2012). Tomato fruits overexpressing the pepper GH3 gene
show anticipation of ripening (Liu et al., 2005), which is in agreement with
the view that the ratio between IAA and its AA-conjugated form, rather than
the level of IAA itself, may contribute to the temporal regulation of ripening
(Bottcher et al., 2010). In nonclimacteric fruits, no single growth regulator
appears to play a positive role analogous to that played by ethylene, but it
has been observed that auxin can negatively control the ripening of some
nonclimacteric fruits. In strawberry, it has been shown that the expression
of many ripening-specific genes can be downregulated by treatments with
an exogenous auxin. Also, in grape, auxin seems to play a negative role in
the regulation of ripening, with synthetic auxin treatments delaying the
expression of a number of ripening-related genes (Davies et al., 1997).

10.3.3 Gibberellins
As a consequence of the prominent role of auxin in the development and
ripening of some nonclimacteric fruits, little attention has been paid to pos-
sible roles of other plant hormones, such as gibberellins (GAs). However,
in strawberry, it has been reported that external application of GA 3 to
­ripening fruits caused a significant delay in the development of the red
color (Martinez et al., 1996) and modified the expression of genes involved
in cell enlargement (de la Fuente et al., 2006) and cell wall disassembly
(Bustamante et al., 2009).

10.3.4  Abscisic Acid

In plants, the phytohormone abscisic acid (ABA) is known to be involved
in various aspects of plant growth, development, and responses to environ-
mental stresses (Leung and Giraudat, 1998; Finkelstein and Rock, 2002;
Himmelbach et al., 2003; Hirayama and Shinozaki, 2007). ABA promotes

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sugar accumulation in fleshy fruits (Yamaki and Asakura, 1991; Kobashi

et al., 1999; Richings et al., 2000; Pan et al., 2005) and plays a role in the
regulation of climacteric and nonclimacteric fruit ripening (Coombe, 1992;
Davies et al., 1997; Giovannoni, 2001; Rodrigo et al., 2003; Zhang et al.
2009a; Sun et al., 2012). In tomato, the suppression of the gene that cata-
lyzes the first step in ABA biosynthesis (NCED1, 9-cis-epoxycarotenoid
dioxygenase) results in the downregulation of some ripening-related cell
wall genes, such as polygalacturonase and pectinmethylesterase, as well
as an increase in firmness and longer shelf life (Sun et al., 2012). Similarly,
reduction of NCED expression correlates with retardation of ripening in
strawberry (Jia et al., 2011). ABA is considered a ripening inducer in straw-
berry and grape fruits (Chai et al., 2011; Jia et al., 2011). The mechanisms
of ABA signaling are not known; however, in grape, analysis of the GH3 pro-
moter identified ABRE-like elements, which may indicate that the ABA–
auxin content ratio is related to the initiation of ripening (Perkins-Veazie,
1995; Jiang and Joyce, 2003; Bottcher et al., 2010).
In recent years, the level of understanding of the molecular events at
the transcriptional, biochemical, hormonal, and metabolite levels underly-
ing ripening in climacteric and nonclimacteric fruits has increased consid-
erably. However, we still poorly understand the developmental switch that
occurs in hormone responsiveness during the transition from immature to
ripe fruits. To date, most published studies of transcriptional and metabolic
regulation are of relatively low resolution at both the spatial and temporal
levels and are furthermore restricted in coverage of various cell molecular
entities. However, new emerging technologies, as well as improved statisti-
cal tools (Klie et al., 2011), allow us to further refine our analytical ability
in order to cope with issues such as subcellular compartmentation and con-
trasting behavior of different cell types (Caldana et al., 2012). Additionally,
the availability of high-quality fruit genome sequence data (Jaillon et al.,
2007; Shulaev et al., 2011; Tomato Genome Consortium, 2012) will aid our
understanding of the genetic regulation of fruit development and ripening.

10.4  Epigenetic Regulation of Fruit

Development and Ripening
Epigenetic regulation of gene expression (inheritance without an alteration
in the primary DNA sequence) is increasingly recognized as a mechanism
for modulating genome activity. Naturally occurring epigenetic changes at
a single gene locus in plants can result in heritable morphological variation
without alteration of the underlying DNA sequence (Patterson et al., 1993;
Cubas et al., 1999; Manning et al., 2006). DNA methylation is one form of
epigenetic regulation. It is involved in transcriptional regulation and stress

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responses and furthermore plays a major role in protecting the genome

integrity against the activity of transposable elements (TEs) and other
repetitive sequences (Chan et al., 2005).
In plants, DNA methylation occurs at cytosine residues in three dif-
ferent sequences (CG, CHG, and CHH, where H = A, C, or T) (Cokus et al.,
2008) and is set in place and maintained by different factors (Law and
Jacobsen, 2010). Analysis of epigenetic variation in Arabidopsis revealed that
at least one-third of expressed genes are methylated in their coding region,
and only 5% of genes are methylated within promoter regions (Zhang et al.,
2006; Vaughn et al., 2007). However, the promoter-methylated genes have a
higher degree of tissue-specific expression (Zhang et al., 2006; Zilberman
and Henikoff, 2007).
The first survey of the frequency and distribution of cytosine methyla-
tion sites in tomato dates back to more than 20 years ago, when it was found
that polymorphisms in cytosine methylation between two tomato species
were relatively abundant, and that methylation patterns were stably inher-
ited, from parents to offspring, segregating in a Mendelian fashion. The
presence of tissue-specific methylation patterns and the overall decrease
of 5-mC frequency in developing tissues also led the authors to postulate
variation of methylation status of selected alleles during plant development
(Messeguer et al., 1991).
More recently, the impact of cytosine methylation on tomato fruit rip-
ening has strikingly emerged in the definition of the molecular nature of
the colorless nonripening phenotype. The tomato nonripening Cnr mutant
fails to produce ripe berries; fruits exhibit green pericarps and do not
respond to external applications of ethylene. The gene at the Cnr locus was
identified as SPB protein–like using positional cloning, but the nonripen-
ing phenotype could not be attributed to any alteration in the coding gene
sequence. Bisulfite sequencing of the Cnr mutant allele showed instead
hypermethylation of cytosine in the region upstream of the predicted ATG
start site. This hypermethylation state correlated with a drastic reduction
of Cnr gene expression (Manning et al., 2006). Therefore, the nonripen-
ing phenotype was due to the heritable cytosine hypermethylation pattern
of the region, including the Cnr gene promoter. Additionally, in normal
tomato fruit (cv. ‘Liberto’) development, the promoter of Cnr appears to be
demethylated in a specific region just prior to the onset of ripening. This
led to the hypothesis that DNA methylation contributes to the regulation of
fruit ripening (Seymour et al., 2008).
Recent work by Zhong et al. (2013) provides genome-wide insights
into the link between the genetic program of fruit ripening and the DNA
methylation state. On the basis of the previous results on the nature of the
Cnr (epi) mutation, the authors injected a chemical inhibitor of cytosine
methylation, 5-azacytidine, directly in the locular spaces and columella of
developing tomato fruits. The methylation inhibitor induced the formation

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of local ripe areas, red in appearance, where the expression of typical

­r ipening-related genes (phytoene synthase 1 and p ­ olygalacturonase) was
anticipated. Moreover, the Cnr promoter region was demethylated in red
sectors with respect to green parts of the fruits, pointing at the demeth-
ylation of Cnr as the epigenetic signal sufficient to induce ripening. The
authors then extended their views on the role of cytosine methylation,
reporting the full tomato methylome sequences of leaves and immature
and ripe fruits, including the ripening-impaired mutants Cnr and rin.
The sequencing of the entire epigenome revealed at least three impor-
tant results: (1) in wild-type fruits, the degree of methylation of regions
upstream of the transcription start sites (TSSs) decreased gradually along
fruit development; (2) this general decline was not observed for the fruits
of the ripening-impaired mutants Cnr and rin, whose CG methylation lev-
els were constantly higher at TSSs and, for Cnr, also comparable to those
observed in leaves; and (3) the promoters of typical ripening-related genes
were gradually demethylated during the development of wild-type fruits.
Further evidence about the link between ripening and cytosine methyla-
tion came from the ChIP-Seq mapping of RIN binding sites during fruit
development. The set of RIN targets included 292 genes with a known role
in ripening. RIN binding sites were found to be adjacent to or overlapping
the methylation hot spots upstream of the TSSs. The analysis of methyla-
tion status of these regions showed that they were progressively demeth-
ylated during the transition from green to red ripe fruits, and this lower
level of methylation correlated with higher transcript levels of RIN target
genes. A previous study showed that the binding of RIN to a limited set of
promoters was inhibited in the Cnr epimutant, indicating that promoter
hypermethylation may prevent RIN binding (Martel et al., 2011). These
three main findings—that (1) local treatment of immature fruits with a
DNA demethylating chemical accelerates ripening; (2) promoters of ripen-
ing genes, which contain RIN binding sites, are gradually demethylated
during ripening but remain stably hypermethylated in ripening-deficient
mutants; and (3) RIN does not bind hypermethylated Cnr gene promoters
(and, possibly, all hypermethylated promoters of its target genes)—taken
together, assign a key role to the epigenome structure and developmen-
tal dynamics in coordinating tomato fruit ripening. The global scenario
presented so far also suggests that progressive demethylation of ripening-
related gene promoters may be the necessary condition for binding of tran-
scriptional regulators, thus triggering the accumulation of ripening-related
transcripts. In normal fruit development, however, the mechanism induc-
ing demethylation of promoters remains elusive, and further efforts are
needed in this direction to uncover the “missing link.” Given the growing
importance of epigenetic modifications in impacting fruit phenotypes, it
is envisaged that, in the future, high-throughput sequencing technologies
will allow routine screening of crop epigenomes, accelerating detection of

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epigenetic variation. It is hypothesized that screening epigenome struc-

ture and dynamics will coexist with the analysis of conventional genetic
variation in future plant breeding strategies. Epigenetic-based crop
­
improvement approaches may radically impact fruit quality traits, espe-
cially for those traits whose allelic variation has been reduced during
domestication or recent intensive breeding pressure. As such, future mod-
eling work aimed at integrating epigenomic profiling and small RNA pro-
filing alongside the more frequently used transcript, protein, enzyme, and
metabolite profiling will allow far greater understanding of the complex
dynamics underlying this tightly regulated biological process.

10.5  Ripening Pathways and Associated

Fruit Quality
The progression of fruit ripening or senescence is a complex process
involving changes to the metabolic and physiological traits of a fruit. In all
fruit, in the tissue surrounding the seed, there is a color change and a change
in cell wall composition causing either a dehiscence or a softening (Klee and
Giovannoni, 2011). Unique to fleshy fruit, there is often a breakdown of stored
carbohydrates to sugars and a decrease in acidity, along with an increase in fla-
vor and aroma volatiles (Klee and Giovannoni, 2011). The control of ripening
appears to be achieved predominantly through the ripening hormones ABA
and ethylene (Fedoroff, 2002; Giovannoni, 2004; Setha, 2012), with ethylene
being the most studied. Fruit types that have a strong requirement for eth-
ylene to ripen, such as tomatoes, peaches, bananas, apples, and melon, have
previously been labeled climacteric, and the role of ethylene in both these
fruit types has been extensively reviewed (Bapat et al., 2010; Paul et al., 2012).
In peaches and tomato, indole-3-acetic acid (IAA) has also been reported
to have some cross talk with ethylene during ripening as (1) production of
ethylene can be concomitant with an increase of IAA and (2) auxin signal-
ing components can be upregulated by ethylene and vice versa (Jones et al.,
2002; Trainotti et al., 2007). In fruit that have a lower requirement of ethylene
to ripen (referred to as non­climacteric fruit such as grape and citrus), ABA
appears to have a stronger role (Setha, 2012). It has been shown that in the
climacteric fruits tomato and banana, there is an increase in ABA preced-
ing an increase in ethylene. Exogenous application of ABA induces ethylene
through the biosynthesis genes (Jiang et al., 2000; Zhang et al., 2009b), while
suppression of ABA leads to a delay in fruit ripening (Sun et al., 2012). In the
dry dehiscent fruit Arabidopsis, again ABA increases with silique maturation
(Kanno et al., 2010) and has been linked with the promotion of dehiscence,
an ­ethylene-mediated event (Child et al., 1998; Kou et al., 2012). While there
is a considerable amount of literature on fruit ripening, researchers have
often only focused on a small number of physiological changes to document

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the ripening process. For example, color change and fruit firmness are often
used as a surrogate for ripening, with other ripening characters completely
overlooked. It is becoming clear that some ripening traits are controlled inde-
pendently of each other (Johnston et al., 2009; Ireland et al., 2013). The use
of single physiological markers may hence lead to a misrepresentation of this
complex process. Here we have summarized the literature based on how dif-
ferent traits respond to hormones, rather than considering ripening as one
single process.

10.5.1  Sugar Accumulation

There is little literature on the hormonal control of starch hydrolysis and the
resulting sugar accumulation. There have been a number of studies that
have documented the metabolic changes that occur during maturation
and ripening (Fait et al., 2008; Osorio et al., 2011, 2012), though the link
between hormonal control and metabolite accumulation is limited; how-
ever, Johnston et al. (2009) observed in apple that, while this could progress
independently of ethylene, it was highly sensitive to ethylene. In melon, the
application of exogenous ABA was shown to promote starch hydrolysis (Sun
et al., 2012), different from growth section; however, this was confounded
by the fact that the ABA also increased the ethylene levels.

10.5.2  Cell Walls and Fruit Shelf Life

The cell wall has a profoundly important influence on fruit texture and cell
wall components, and the underlying genes have been frequent targets for
genetic engineering, mostly in tomato, with the goal of extending shelf life
(Vicente et al., 2007; Goulao and Oliveira, 2007). Although knowledge of
the mechanism of fruit softening has grown in recent years, it has proven
especially difficult to establish the relationship between specific aspects
of cell wall metabolism or architecture and their relationship to changes
in tissue firmness (Vicente et al., 2007). Similarly, it has also been difficult
to establish the fine regulation of genetic elements that produce a range
of effects depending on the genetic background in which they are intro-
duced (Faria et al., 2006). The regulation of texture and shelf life is clearly
far more complex than was previously envisaged, and so new approaches
are needed, including the inclusion of observations in species beyond the
traditional tomato experimental model (Ezura and Owino, 2008); a better
understanding of the relationship between changes in the textural proper-
ties of specific fruit tissues, intact fruit firmness, and shelf life; and more
comprehensive models of the biochemical and physiological elements that
contribute to fruit firmness.

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Ripening-related disassembly of the cell wall polysaccharide matrix is

generally the only factor that is mentioned when describing the structural
basis of fruit softening and the associated loss of shelf life and fruit quality.
This is largely a reflection of the relative ease with which some of the dra-
matic changes in wall polymer structure and composition, and the activi-
ties of the associated enzymes, can be measured. However, recent studies
have started to examine the potential involvement and relative importance
of other structures and physiological processes. For example, it has been
proposed that differences in tomato fruit cuticle structure and composi-
tion may be associated with the substantial variation in tomato fruit shelf
life that has been reported in different tomato genotypes (Saladie et al.,
2007). The cuticle has a number of biological functions that could have an
important impact on fruit quality and shelf life, including the ability to main-
tain fruit skin integrity (Hovav et al., 2007), restrict cuticular transpiration
(Leide et al., 2007), and limit microbial infection.
Other reports have also highlighted ripening-related processes that
likely contribute to fruit firmness, such as turgor pressure (Saladie et al.,
2007) and the possibly associated developmental changes in apoplastic sol-
ute accumulation (Wada et al., 2008). Integration of all these features into
a more holistic model of ripening-related textural changes should provide a
host of new targets for gene manipulation, with the ultimate goal of allowing
more complete fruit ripening on the plant, thereby promoting nutrient and
flavor accumulation while maintaining fruit firmness and shelf life.
Depending on the fruit type, the cell wall can manifest as a formation
of a dehiscence zone or through the softening of the flesh tissue. In each
case, there is a suite of cell wall–related genes that are upregulated, and in
many instances, each is differentially regulated. In the case of fruit soft-
ening, the loss of a single gene can be compensated by other gene action
(Powell et al., 2003). In apple and melon, there is both ethylene-independent
and ethylene-dependent softening, which can be observed in the differen-
tial regulation of cell wall–related genes. In banana, it has been shown that
ABA can act synergistically with ethylene to promote softening (Lohani
et al., 2004), and in grape ABA has been shown to cause fruit softening
(Cantin et al., 2007).

10.5.3  Color, Flavor, and Nutrition in Fruits

10.5.3.1  Chloroplast-to-Chromoplast Conversion

One of the most critical developmental acts associated with ripening and
palatability is the conversion of chloroplasts to chromoplasts (Egea et al.,
2010). There are a number of important structural changes that accom-
pany chromoplast conversion. These changes have a major influence on the

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nutrient and flavor composition of the fruit. The photosynthetic capacity of

the chloroplast is lost as the thylakoid structures begin to disassemble.
Within the chromoplast, plastoglobules accumulate. These are the sites
for accumulation of large quantities of carotenoids, principally lycopene
and β-carotene, in the form of crystal structures. Accumulation of these
carotenoids provides a visual indication that the fruit is mature and suit-
able for consumption. These carotenoids are also important human nutri-
ents, providing the precursor of vitamin A as well as abundant antioxidants
to the diet.
The chromoplast and plastoglobule proteomes have been examined
in multiple species. The tomato chromoplast is highly metabolically active,
and enzymes associated with synthesis and metabolism of carotenoids,
amino acids, and fatty acids are detected (Barsan et al., 2010). Although the
tomato plastoglobule proteome has not been reported, that of the closely
related pepper has been determined (Ytterberg et al., 2006). As expected,
the enzymes responsible for carotenoid synthesis are detected within these
structures. The conversion of chloroplasts to chromoplasts and associated
degradation of chlorophyll and membrane structure–photosynthetic activ-
ity with concomitant carotenoid accumulation are the main hallmarks of
the ripening transition of tomato (and many fruits) and are irreversible. The
primary enzymatic regulator of flux into the carotenoid pathway, phytoene
synthase (PSY), is under strong positive ethylene control during ripen-
ing, indicating a link between chromoplast development and the primary
­r ipening hormone. Also under ethylene control is repression of the gene
encoding the lycopene β-cyclase (LYC) enzyme that would otherwise con-
vert red lycopene to orange β-carotene (Alba et al., 2005). Few regulatory
genes or activities specifically involved in this process beyond those involved
in carotenoid synthesis and chlorophyll degradation (chlorophyllase) have
been described. One gene clearly involved in this transition is defined by
the green-flesh (gf ) locus that encodes a Stay-Green senescence-related
regulator (Barry et al., 2008). The gf loss-of-function mutations result in
reduced plastid conversion during ripening, yielding fruit containing both
chloroplasts and chromoplasts and characterized by a brown or chocolate
color. Mutation in the orthologous pepper gene has the same effect (Barry
et al., 2008; Borovsky and Paran, 2008), and such fruits are used commer-
cially because of their novel appearance and flavor characteristics.

10.5.3.2  Flavor and Aroma Production

In apple, aroma volatiles are the least ethylene sensitive and most ethylene
dependent of the ripening traits. Consistent with this, there are a significant
number of publications linking the production of aroma with ethylene (Flores
et al., 2002; Botondi et al., 2003; Defilippi et al., 2005; Schaffer et al., 2007).

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There is, however, remarkably little literature examining whether other

­hormones contribute to the regulation of volatile production in fruit.

10.5.3.3  Control of Color and Texture Changes

Much of the literature documents the control of color change during fruit
ripening. This is achieved by a combination of chlorophyll loss (degreen-
ing) and production of secondary color metabolites such as carotenoids
and anthocyanins. Color change in many fruit species is associated with
an increase of ABA or ethylene. In apple, the degreening occurs inde-
pendently of ethylene, but ethylene can accelerate the process (Johnston
et al., 2009). Citrus and melon also both require ethylene for the degreen-
ing of the skin. The production of secondary color metabolites is strongly
ethylene regulated in tomato, though some intermediates can be pro-
duced in the absence of ethylene. Application of ABA to tomato fruit
results in an enhanced onset of breaker stage compared to controls, fur-
ther implicating ABA as being a positive regulator of ripening in tomato
(Buta and Spaulding, 1994). In grapes and strawberry, the color change is
strongly regulated by ABA (Deytieux et al., 2005; Jia et al., 2011), though
application of 1-methylcyclopropene (1-MCP) (an inhibitor of ethylene
response) can delay this process, suggesting that ethylene may play a
role (Chervin et al., 2004). There are also reports of color change being
­inhibited by brassinosteroids in grape and strawberry (Symons et al.,
2006; Chai et al., 2013).
A systems biology approach to the tomato transcriptome and metabo-
lome during ripening has led to the identification of a putative carotenoid
modulator, SlERF6 (Lee et al., 2012). Reduced expression of SlERF6 by
RNAi enhanced both carotenoid and ethylene levels during ripening. The
majority of genes substantially influenced by SlERF6 repression were
upregulated, indicating that the primary function of SlERF6 occurs via
negative regulation. SlERF6 was responsive to ethylene and may be a com-
ponent of a feedback restriction in ethylene production during ripening,
similar to SlAP2 (Lee et al., 2012). A small number of fruit high-pigment
mutants have been described in tomato, including high pigment  1 (hp1)
(a  lesion in UV-DAMAGED DNA BINDING PROTEIN1 [DDB1]) (Liu
et al., 2004) and hp2 [a mutation in tomato DE-ETIOLATED1 (DET1)]
(Mustilli et al., 1999). These genes are involved in the suppression of light
responses in the absence of light, and for DET1, suppression occurs by a
molecular mechanism involving chromatin remodeling (Davuluri et al.,
2005). Coordinated upregulation of core metabolic processes is responsible
for the high-­pigment phenotype in the DET1 lines (Enfissi et al., 2010).
Upregulation of a Golden 2-like transcription factor in tomato has also been
shown to elevate levels of chlorophyll and carotenoids, and a lesion in this
gene is responsible for the uniform ripening phenotype (Powell et al., 2012).

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Shelf life is important in tomato and other fruits. This trait is influ-
enced by many processes, including the rate of fruit softening. Modulation
of genes encoding two ripening-specific N-glycoprotein-modifying
enzymes—α-mannosidase and β-D-N acetyl hexosaminidase—reduces
the rate of softening and enhances fruit shelf life (Meli et al., 2010). More
than 50 cell wall structure-related genes show expression changes in
­r ipening tomato fruits (Tomato Genome Consortium, 2012), and texture
involves complex quantitative trait loci (Chapman et al., 2012). Shelf life is
also related to water loss from the mature fruits.
Cuticle properties are critical for maintaining fruit integrity post-
harvest. Delayed Fruit Deterioration (DFD) mutant tomato fruits show
­remarkably prolonged resistance to postharvest desiccation and remain
firm for many months after harvest (Saladie et al., 2007), and analyses of
cutin-deficient tomato mutants indicate the importance of waxes (Isaacson
et al., 2009).

10.6  Human Nutrition and Fruits

The human genome evolved in the context of the diet, prevailing before
the introduction of agriculture and animal husbandry approximately 10,000
years ago, when humans were hunter-gatherers; diets were rich in fruits,
vegetables, and protein and low in fats and starches (Eaton and Konner,
1985). For primates such as chimpanzees and orangutans, more than 75%
of their diet by weight is fresh fruit, and the assumption is that our primate
ancestors had similar diets. Fruit is also common in the diet of modern
hunter-gatherers. The 10,000 years (∼360 generations) since the first culti-
vation of cereals has not been enough time for humans to adapt to starchier,
cereal-based diets with much higher amounts of fat and lower amounts of
fresh fruit and vegetables, and it has been suggested that many chronic dis-
eases result from our evolutionary discordance with modern diets (Cordain
et al., 2000). Levels of phytonutrient compounds in plant-based foods that
play potentially beneficial roles in the prevention and treatment of disease
in the diet have dropped significantly owing to a reduced variety of plants
consumed (currently, just 17 plant species constitute 90% of the global
human diet) and selective breeding (Willett, 2010). For these reasons, most
modern dietary recommendations include increased consumption of fresh
or whole fruits and vegetables. The role of plants in human health is the
subject of another review (Martin et al., 2013), but what is beneficial about
fruit consumption?
Fruits such as eggplant, mango, and okra are good sources of soluble
fiber, which has been shown experimentally to lower the glycemic index
of foods and have beneficial effects on type 2 diabetes, cardiovascular dis-
ease, and obesity (Larsson et al., 2009). Fruits are also rich in fructose

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(which constitutes 20%–40% of the available carbohydrates in wild fruit)

and ­intrinsic sugars; these are not subject to restrictions in dietary rec-
ommendations because their concentrations are not high enough to have
adverse health outcomes (Mann, 2004). Fruits generally also contain high
levels of phytonutrients, and include carotenoids, polyphenols (flavonoids,
stilbenes), plant sterols, vitamins, and polyunsaturated fatty acids.
Some fruits are rich in carotenoids, which are thought to attract
­a nimals as dispersers through their bright colors (e.g., lycopene in tomato;
capsanthin, β-carotene, lutein, and violaxanthin in red pepper; lycopene
and β-carotene in red grapefruit; and β-carotene, α-carotene, and lutein
in mangoes). Lycopene is a methyl-branched carotenoid that has no provi-
tamin A activity. It is a potent lipophilic antioxidant, with greater antioxi-
dant activity than other carotenoids, and has been shown to be protective
against cardiovascular disease and prostate cancer (Fraser and Bramley,
2004). Intervention and epidemiological studies have linked β-carotene con-
sumption to enhanced protection against cardiovascular disease, including
cerebral infarction, and resistance to low-density lipoprotein oxidation,
ischemic stroke, and carotid atherosclerosis (Fraser and Bramley, 2004).
Fruits are rich sources of polyphenols that have gained significant
recognition recently as being important phytonutrients, reducing the risk
of chronic diseases such as cardiovascular disease, metabolic syndrome,
cancer, and obesity. Anthocyanins represent a subset of flavonoids with
particularly high antioxidant capacity and strong health-promoting effects.
As part of the human diet, they offer protection against cancer, inhibiting
both the initiation and progression stages of tumor development (Tsuda
et al., 2003), reducing inflammatory inducers of tumor initiation, suppress-
ing angiogenesis, and minimizing cancer-induced DNA damage in animal
disease models. Anthocyanins also protect against cardiovascular disease
and age-related degenerative diseases associated with metabolic syndrome
(Hou et al., 2004), have anti-inflammatory activity, promote visual acuity,
and hinder obesity and diabetes. Inverse correlations between consump-
tion of flavonol-rich diets and the occurrence of cardiovascular disease,
certain cancers, and age-related degenerative diseases have been shown
in human epidemiological studies as well as by data from cell-based assays
and feeding trials with animals.
Another important group of plant-based bioactive polyphenols is the
hydroxycinnamic acid esters, of which chlorogenic acid is the major soluble
phenolic in solanaceous species such as potato, tomato, and eggplant, as
well as in coffee. Chlorogenic acid is one of the most abundant polyphenols
in the human diet and is the major antioxidant in the average developed
world diet. It has significant antioxidant activity, and consumption can limit
glucose absorption and possibly weight gain (Thom, 2007).
Epicatechins are the major polyphenolic compounds in green tea, and
the most significant active component is thought to be epigallocatechin

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gallate. Cell, animal, and human studies have shown that green tea extract
and epicatechins prevent cancer development, particularly prostate can-
cer (Dixon and Ferreira, 2002). Similar effects may be gained by the
­consumption of fleshy fruits such as cranberry, blueberry, and grape or by
the consumption of berry-juice products; these have relatively high levels
of catechins and epicatechins, which also protect against cardiovascular
disease (Dell’Agli et al., 2004).
Dietary intervention studies support the view that consumption
of fruit (especially when replacing calorie-dense food) helps maintain a
healthy weight and reduce the risk of a wide range of cancers (and car-
diovascular disease). Consequently, dietary improvement by increasing
fruit consumption or by improving fruit to contain higher levels of fiber or
phytonutrients should have a positive impact on the incidence and progres-
sion of chronic disease. Such improvement of fruit crops could be carried
out through genetic modification (Butelli et al., 2008) or marker-assisted
­selection using existing variation.
Fresh fruits and their processed derivates represent important
sources of carotenoids, flavonoids, and anthocyanins, and the possibility
of improving content using bioengineering represents an opportunity to
improve access to healthy foods. The inherent resiliency of plant metab-
olism to maintain homeostasis has impaired efforts to facilitate changes
in some of these pathways, as has been especially well documented for
carotenoids (Fraser et al., 2009). An alternative approach is to focus on
regulatory rather than biosynthetic genes. Accumulation of carotenoids in
high-pigment fruit mutants has been reported to result from an increase in
the plastid number, larger plastid compartment size, increased plastid divi-
sion, and greater capacity for pigment storage (Galpaz et al., 2008; Wang
et al., 2009). These changes in pigmentation, plastid type, and metabolism
were associated with the elevation of transcripts from key carotenoid genes
such as phytoene synthetase 1 (Psy-1) and are not directly connected with
the ripening process (Fraser et al., 2007), thus allowing modifications
in carotenoid content without affecting other aspects of fruit ripening or
broader plant development. Moreover, this particular modification has
been reported to be stable in field tests for transgene stability and yield
performance (Giorio et al., 2007).
A MYB transcription factor from snapdragon was expressed using
the E8 promoter specifically in tomato fruit, causing elevated levels of
anthocyanins, compounds that have been shown to increase the longev-
ity of cancer-susceptible mice (Butelli et al., 2008). A homolog of this
transcription factor (MYB10) was previously cloned and characterized
from apple and was shown to positively regulate the anthocyanin pathway
(Espley et al., 2007). Further, an allelic rearrangement in the promoter of
MYB10 causes a novel autoregulatory motif, which is sufficient to account
for the increase in MYB10 levels and subsequent ectopic accumulation of

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anthocyanins throughout the plant (Espley et al., 2009). This modification

was found in all red apple species tested, but not in white apple varieties.
This gain-of-function mutation in the anthocyanin regulatory pathway has
significant implications for the development of novel varieties of plants and
fruit with enhanced nutritional status and increased consumer appeal, by
using either genetic modification or more conventional breeding methods.
There are lots of examples of specific pathways that have been tar-
geted through genetic engineering with either enhanced shelf life or
­nutritional status, or genotypes of particular interest that are known today.
However, impacts on health and nutrition can be mediated through less
direct means than the alteration of nutrition-associated biochemical path-
ways. These goals can be met by modifying overall appeal and quality so
that fruit and fruit products become more competitive choices for consum-
ers and net fruit consumption is increased. For example, seedless tomato
fruits were developed by silencing a key enzyme of the flavonoid biosynthe-
sis pathway (Schijlen et al., 2007).

10.6.1  Human Nutrition: Functional

Genomics/Systems Approach
Prior efforts toward fruit quality improvement have focused on single steps
in biochemical pathways, but complex feedback regulation networks have
complicated such strategies. Targeting regulatory genes may provide a
route to avoid these regulatory impediments, though the difficult work of
assessing the impact of altering specific pathway steps is required to under-
stand the underlying regulatory networks (Fraser et al., 2007). The use
of a transcription factor that regulates a complete biosynthesis pathway,
as opposed to targeting a single enzyme, has proved useful to study and
manipulate the accumulation of tannins in grape (Bogs et al., 2007) or poly-
phenolic antioxidants in tomato (Luo et al., 2008). New high-throughput
functional genomics and systems approaches are also allowing more accu-
rate predictions of regulatory processes (Phillips, 2008).
Applying such methodologies to mapping of quantitative trait loci
(QTL) allowed for the detection of epistatic interactions among genes
(Causse et al., 2007), the discovery of new genes related to metabolic
pathways of interest, and even testing of the stability of QTL effects over
­multiple seasons and in different environments (Stevens et al., 2007).
Recent reports revealed that the screening for genes whose
expression varies between lines that are genetically very similar and
yet s­ ignificantly different in fruit quality is a helpful tool for identifying
potentially valuable genetic targets (Obando et al., 2008) and provides
insights concerning the nature of complex genetic control processes

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that relate to fruit nutrient content (Cuevas et al., 2008), ripening, and
softening (Moreno et al., 2008). These tools make it possible to identify
potential candidate genes in model plants, like Arabidopsis, which could
lead to targeting important genes in crop species (Gorguet et al., 2008).
In one interesting example, tomatoes were biofortified with calcium by
the overexpression of an Arabidopsis calcium transporter (CA X1) (Park
et al., 2005). However, while the calcium levels were increased, the bio-
availability of the increased calcium was questioned. Similar experiments
were carried out, where the intake of calcium from transgenic carrots
overexpressing a CA X1 gene was monitored in both mice and humans
(Morris et al., 2008). Calcium absorption in bones increased for both
mice and humans eating CA X1-expressing carrots compared to controls,
demonstrating alternative means of fortifying fruits and vegetables with
­bioavailable calcium.

10.7  Horticultural Crop Improvement

High-throughput sequencing technologies have provided a powerful new
set of tools to reveal genetic and epigenetic variation on a genome-wide
scale, and therefore new insights into the evolution of genomes and trait
variation. Fruit crop genomes that have been sequenced include grapevine
(Vitis vinifera), apple (Malus × domestica), diploid strawberry (Fragaria
vesca), tomato (Solanum lycopersicum), and banana (Musa acuminata).
These resources will underpin advances in the breeding of fruit crops by
(1) greatly facilitating marker-assisted selection and allowing efficient clon-
ing of genes underpinning quantitative trait loci, (2) providing the refer-
ence genomes for rapid allele mining in crop wild relatives (Lippman et al.,
2007), (3) providing a guide for direct sequencing of monogenic mutants,
and (4) maximizing opportunities to optimize reverse-genetics tools, such
as targeting-induced local lesions in genomes (TILLING) for gene func-
tional studies (Stephenson et al., 2010).
Genome-wide information also allows rapid analyses of gene struc-
ture, binding site motifs, and gene regulatory regions. In the future, high-
throughput sequencing technologies will allow routine screening of crop
epigenomes, and therefore permit detection of epigenetic variation that
impacts phenotypes. Additionally, translational biology from models to
crop species has clear utility in enhancing important crop traits; for exam-
ple, fine-tuning the expression of the value margin identity gene IND in
Brassica gives a potentially useful partial dehiscence phenotype (Girin
et al., 2010). Harnessing variation in crop wild relatives in combination with
direct genetic modification provides a powerful route for a massive step
change in horticultural crop improvement.

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10.8  Genetic Manipulation of

Ripening Regulatory Genes
To date, attempts to manipulate the expression of target genes during fruit
ripening have produced varying results. As mentioned, transcriptional con-
trol has been utilized with respect to breeding for heterozygosity at the
rin mutation (Rin/rin) in commercial lines to delay ripening and extend
shelf life; however, this modification leads to the inhibition of flavor and
nutritional compound accumulation, along with undesirable textural traits
in some backgrounds due to incomplete ripening. In addition, constitutive
expression or silencing of other target genes can cause unwanted pleio­
tropic effects on other aspects of plant development, or even lead to lethal-
ity, as is probably the case for the Cnr and LeHB-1 loci, as stable transgenic
plants harboring either of these genes have been difficult to produce. As
a consequence of unwanted pleiotropic effects, targeted analysis of the
transgene effect in the fruit is often difficult, as exemplified by the light
signal transduction and carotenoid regulatory gene HIGH-PIGMENT 2,
where pleiotropic effects of transgene expression in nonfruit tissues were
noted (low yield and brittleness), although this can be overcome through
the use of a fruit-specific promoter (Davuluri et al., 2005). More accurate
phenotypic determination of fruit function and utility for the assessment
of other production or quality influences will be achieved through the use
of appropriate tissue-specific and development-specific promoters. Indeed,
the most useful transgenic or natural alleles for ripening control will be
those that deliver fruit specificity and result in levels of expression suffi-
cient for extended shelf life while ripening sufficiently for desirable quality.
While several fruit-specific promoters are known, the commercial use of
most is subject to intellectual property restrictions, and even they repre-
sent a narrow range of expression options (essentially either throughout
fruit development or induced at the onset of ripening). Isolation and uti-
lization of promoters with more desirable temporal or spatial expression
profiles could provide the means to regulate fruit ripening by manipulating
the expression of transcription factors such as rin, Cnr, and LeHB-1, absent
unintended effects. Further, more detailed characterization of the molecu-
lar makeup of ripening regulatory networks (Alba et al., 2005) should pro-
vide additional fruit-specific loci that can be altered genetically to avoid
undesirable secondary effects, without the need of fruit-specific promoters.

10.9  Conclusions and Future Perspectives

The ethylene response that defines a climacteric fruit coordinates the
expression of thousands of genes that in turn control fruit softening as
well as accumulation of pigments, sugars, acids, and volatiles. Together,

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this network of transcriptional and hormonal regulators mediates the

­physiological changes that make the fruit attractive to seed-dispersing
organisms and, with the availability of a high-quality tomato genome
sequence, is the foundation for expansion and refinement of our under-
standing of the genetic regulation of fruit development and ripening.
It is important to note that recent studies have reported that geneti-
cally modified organisms are not inherently dangerous and can be consid-
ered safe to consumers when compared with conventional foods developed
for target traits through naturally occurring genetic variation (Venneria
et al., 2008). Of course, given that the safety of crops is a primary con-
cern for both producers and consumers, it is necessary to define the risk
and unintended effects in genetically modified plants (Deng et al., 2008),
including the effects of a specific gene and the possible effects in a sus-
ceptible subgroup of consumers (Finamore et al., 2008). Although many of
the target genes that result from emerging genomics approaches may not
in themselves be candidates for direct manipulation, they may represent
DNA sequences that can be exploited as markers in breeding programs
to select for allelic variation, which can be tested and potentially used for
the alteration of target traits. Selection of QTLs in particular can be made
much more efficient through marked-assisted selection (MAS). Integrating
MAS in traditional breeding practices is most likely the highest short-term
impact of our rapidly increasing understanding of fruit molecular biology,
as it is economically attractive and applicable for markets in both devel-
oped and developing countries (Hospital, 2009). The current proliferation
of genetically modified cereal and fiber crops is likely to alleviate consumer
concerns, reduce the current regulatory costs, and result in more direct
biotechnology applications to fruit crops in coming years.
Some aspects need to be considered for future studies related to fruit
ripening:

• What additional transcriptional regulators remain to be identified,

and what is the relationship of these regulators to each other?
• Are there specific subregulators in terms of transcription factors
or other genes that specifically regulate individual ripening path-
ways or processes distinct from the more pleiotropic regulators
defined to date?
• Can predictive models of the ripening regulatory network be gen-
erated for tomato and other fleshy fruits? And can major ripening
processes, such as texture and color development, be separated to
allow better control over the processes?
• One of the best-characterized natural epigenetic mutations was
discovered in tomato. To what extent is epigenetic variation respon-
sible for controlling important quality traits in crops and crop wild
relatives?

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Epigenetic regulation of gene expression is increasingly recognized as

a normal and essential mechanism for coordinating genome activity in
eukaryotes and thus regulating many aspects of development or response
to the environment (Seymour et al., 2008). Few reports have been published
related to epigenetic regulation, although in Arabidopsis one-third of the
expressed genes are methylated in parts of their coding region, while 5%
are methylated within their promoter regions (Zhang et al., 2007; Seymour
et al., 2008). A new important aspect to gene regulation involves micro­
RNAs (miRNAs). A limited set of miRNAs has been identified in tomato
fruits (Pilcher et al., 2007); they are believed to be fine-tuning mechanisms
in gene regulation (Seymour et al., 2008). Misexpression of miRNAs can
produce pleiotropic effects on development. It would be no surprise that
several of the events related to fruit ripening were under the control of
miRNA expression.
Fruits have been selected as one of the vehicles to human immuni-
zation based on the knowledge related to the biology of the fruits; several
techniques for plant vaccines have been developed, and a few of them
are on their way to the market. Up to now, other biological products have
been produced on fruits (Zhi et al., 2007; Artzen, 2008), which opens
a new perspective for the economic and scientific importance of these
products.

References
Alba, R., Payton, P., Fei, Z., McQuinn, R., Debbie, P., Martin, G.B., Tanksley,
S.D., and Giovannoni, J.J. 2005. Transcriptome and selected metabolite
analyses reveal multiple points of ethylene control during tomato fruit
development. Plant Cell 17, 2954–2965.
Artzen, C.J. 2008. Using tobacco to treat cancer. Science 321, 1052–1053.
Bapat, V.A., Trivedi, P.K., Ghosh, A., Sane, V.A., Ganapathi, T.R., and Nath, P.
2010. Ripening of fleshy fruit: Molecular insight and the role of ethyl-
ene. Biotechnology Advances 28, 94–107.
Barry, C., McQuinn, R., Chung, M., Besuden, A., and Giovannoni, J. 2008.
Amino acid substitutions in homologs of the STAY-GREEN (SGR) pro-
tein are responsible for the green-flesh and chlorophyll retainer muta-
tions of tomato and pepper. Plant Physiology 147, 179–187.
Barry, C.S., and Giovannoni, J.J. 2006. Ripening in the tomato green-ripe
mutant is inhibited by ectopic expression of a protein that disrupts eth-
ylene signaling. Proceedings of the National Academy of Sciences USA
103, 7923–7928.
Barsan, C., Sanchez-Bel, P., Rombaldi, C., Egea, I., Rossignol, M.,
Kuntz, M., Zouine, M., Latche, A., Bouzayen, M., and Pech, J.C. 2010.
Characteristics of the tomato chromoplast revealed by proteomic anal-
ysis. Journal of Experimental Botany 61, 2413–2431.

324
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Bogs, J., Jaffe, F.W., Takos, A.M., Walker, A.R., and Robinson, S.P. 2007. The
grapevine transcription factor VvMYBPA1 regulates proanthocyanidin
synthesis during fruit development. Plant Physiology 143, 1347–1361.
Borovsky, Y., and Paran, I. 2008. Chlorophyll breakdown during pepper fruit
ripening in the chlorophyll retainer mutation is impaired at the homolog
of the senescence-inducible stay-green gene. Theoretical and Applied
Genetics 117, 235–240.
Botondi, R., De Santis, D., Bellincontro, A., Vizovitis, K., and Mencarelli, F.
2003. Influence of ethylene inhibition by 1-methylcyclopropene on apri-
cot quality, volatile production, and glycosidase activity of low- and high-
aroma varieties of apricots. Journal of Agricultural and Food Chemistry
51, 1189–1200.
Bottcher, C., Keyzers, R.A., Boss, P.K., and Davies, C. 2010. Sequestration of
auxin by the indole-3-acetic acid amido synthetase GH3-1 in grape berry
(Vitis vinifera L.) and the proposed role of auxin conjugation during
ripening. Journal of Experimental Botany 61, 3615–3625.
Bustamante, C.A., Civello, P.M., and Martinez, G.A. 2009. Cloning of the pro-
moter region of betaxylosidase (FaXyl1) gene and effect of plant growth
regulators on the expression of FaXyl1 in strawberry fruit. Plant Science
177, 49–56.
Buta, J.G., and Spaulding, D.W. 1994. Changes in indole-3-acetic acid and
abscisic acid levels during tomato (Lycopersicon esculentum Mill.) fruit
development and ripening. Journal of Plant Growth and Regulation 13,
163–166.
Butelli, E., Titta, L., Giorgio, M., Mock, H.P., Matros, A., Peterek, S., Schijlen,
E.G., Hall, R.D., Bovy, A.G., Luo, J., and Martin, C. 2008. Enrichment
of tomato fruit with health-promoting anthocyanins by expression of
select transcription factors. Nature Biotechnology 26,1301–1308.
Caldana, C., Fernie, A.R., Willmitzer, L., and Steinhauser, D. 2012. Unraveling
retrograde signaling pathways: finding candidate signaling molecules
via metabolomics and systems biology driven approaches. Frontiers in
Plant Science 3, 267.
Cantin, C.M., Fidelibus, M.W., and Crisostoc, C.H. 2007. Application of
abscisic acid (ABA) at veraison advanced red color development
and maintained postharvest quality of ‘Crimson Seedless’ grapes.
Postharvest Biology and Technology 46, 237–241.
Castillejo, C., dela Fuente, J.I., Iannetta, P., Botella, M.A., and Valpuesta, V.
2004. Pectin esterase gene family in strawberry fruit: Study of FaPE1,
a ripening-specific isoform. Journal of Experimental Botany 55, 909–918.
Causse, M., Chaib, J., Lecomte, L., Buret, M., and Hospital, F. 2007. Both
additivity and epistasis control the genetic variation for fruit quality
traits in tomato. Theoretical and Applied Genetics 115, 429–442.
Cevik, V., Ryder, C.D., Popovich, A., Manning, K., King, G.J., and Seymour,
G.B. 2010. A FRUITFULL-like gene is associated with genetic variation
for fruit flesh firmness in apple (Malus domestica Borkh.). Tree Genetics
and Genomes 6, 271–279.

325
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Chai, Y.M., Jia, H.F., Li, C.L., Dong, Q.H., and Shen, Y.Y. 2011. FaPYR1 is
involved in strawberry fruit ripening. Journal of Experimental Botany
62, 5079–5089.
Chai, Y.M., Zhang, Q., Tian, L., Li, C.L., Xing, Y., Qin, L., and Shen, Y. 2013.
Brassinosteroid is involved in strawberry fruit ripening. Plant Growth
and Regulation 69, 63–69.
Chan, S.W., Henderson, I.R., and Jacobsen, S.E. 2005. Gardening the genome:
DNA methylation in Arabidopsis thaliana. Genetics 6, 351–360.
Chapman, N.H., Bonnet, J., Grivet, L., Lynn, J., Graham, N., Smith, R., Sun, G.,
Walley, P.G., Poole, M., Causse, M., King, G.J., Baxter, G., and Seymour,
G.B. 2012. High resolution mapping of a fruit firmness-related QTL in
tomato reveals epistatic interactions associated with a complex combi-
natorial locus. Plant Physiology 159, 1644–1657.
Chervin, C., El-Kereamy, A., Roustan, J.P., Latche, A., Lamon, J., and
Bouzayen, M. 2004. Ethylene seems required for the berry develop-
ment and ripening in grape, a non-climacteric fruit. Plant Science 167,
1301–1305.
Child, R.D., Chauvaux, N., John, K., Ulvskov, P., and Van Onckelen, H.A.
1998. Ethylene biosynthesis in oilseed rape pods in relation to pod shat-
ter. Journal of Experimental Botany 49, 829–838.
Chung, M., Vrebalov, J., Alba, R., Lee, J., McQuinn, R., Chung, J., Klein, P., and
Giovannoni, J. 2010. A tomato (Solanum lycopersicum) APETALA2/ERF
gene, SlAP2a, is a negative regulator of fruit ripening. Plant Journal 64,
936–947.
Civello, P.M., Powell, A.L., Sabehat, A., and Bennett, A.B. 1999. An expan-
sin gene expressed in ripening strawberry fruit. Plant Physiology 121,
1273–1280.
Coen, E.S., and Meyerowitz, E.M. 1991. The war of the whorls: Genetic inter-
actions controlling flower development. Nature 353, 31–37.
Cokus, S.J., Feng, S., Zhang, X., Chen, Z., Merriman, B., Haudenschild,
C.D., Pradhan, S., Nelson, S.F., Pellegrini, M., and Jecobsen, S.E. 2008.
Shotgun bisulphate sequencing of the Arabidopsis genome reveals DNA
methylation patterning. Nature 452, 215–219.
Coombe, B.G. 1992. Research on development and ripening of the grape
berry. American Journal of Enology and Viticulture 43, 101–111.
Cordain, L., Miller, J.B., Eaton, S.B., Mann, N., Holt, S.H.A., and Speth, J.D.
2000. Plant-animal subsistence ratios and macronutrient energy estima-
tions in worldwide hunter-gatherer diets. American Journal of Clinical
Nutrition 71, 682–692.
Cubas, P., Vincent, C., and Coen, E. 1999. An epigenetic mutation responsible
for natural variation in floral symmetry. Nature 401, 157–161.
Cuevas, H.E., Staub, J.E., Simon, P.W., Zalapa, J.E., and McCreight, J.D.
2008. Mapping of genetic loci that regulate quantity of beta-carotene
in fruit of US Western Shipping melon (Cucumis melo L.). Theor Appl
Genet 117, 1345–1359.

326
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Davies, C., Boss, P.K., and Robinson, S.P. 1997. Treatment of grape berries, a
non-climacteric fruit with a synthetic auxin, retards ripening and alters
the expression of developmentally regulated genes. Plant Physiology
115, 1155–1161.
Davuluri, G.R., van Tuinen, A., Fraser, P.D., Manfredonia, A., Newman, R.,
Burgess, D., Brummell, D.A., King, S.R., Palys, J., Uhlig, J., Bramley,
P.M., Pennings, H.M.J., and Bouler, C. 2005. Fruit-specific RNAi medi-
ated suppression of DET1 enhances carotenoid and flavonoid content in
tomatoes. Nature Biotechnology 23, 890–895.
Defilippi, B.G., Dandekar, A.M., and Kader, A.A. 2005. Relationship of ethyl-
ene biosynthesis to volatile production, related enzymes, and precursor
availability in apple peel and flesh tissues. Journal of Agricultural and
Food Chemistry 53, 3133–3141.
de la Fuente, J.I., Amaya, I., Castillejo, C., Sanchez-Sevilla, J.F., Quesada, M.A.,
Botella, M.A., and Valpuesta, V. 2006. The strawberry gene FaGAST
affects plant growth through inhibition of cell elongation. Journal of
Experimental Botany 57, 2401–2411.
Dell’Agli, M., Busciala, A., and Bosisio, E. 2004. Vascular effects of wine poly-
phenols. Cardiovascular Research 63, 593–602.
Deng, P., Zhou X., Yang, D., Hou, H., Yang, X., Li, Y., Yang, Y., Wang, X., Fang,
S., Wu, S., and Liu, J. 2008. The definition, source, manifestation of and
assessment of unintended effects in genetically modified plants. J Sci
Food Agric 88, 2401–2413.
Deytieux, C., Geny, L., and Doneche, B. 2005. Relation between hormonal bal-
ance and polygalacturonase activity in grape berry. Acta Horticulturae
682, 163–170.
Dixon, R.A., and Ferreira, D. 2002. Genistein. Phytochemistry 60, 205–211.
Eaton, S.B., and Konner, M. 1985. Paleolithic nutrition—A consideration of
its nature and current implications. New England Journal of Medicine
312, 283–289.
Egea, I., Barsan, C., Bian, W., Purgatto, E., Latche, A., Chervin, C., Bouzayen,
M., and Pech, J.C. 2010. Chromoplast differentiation: Current status
and perspectives. Plant and Cell Physiology 51,1601–1611.
Elitzur, T., Vrebalov, J., Giovannoni, J.J., Goldschmidt, E.E., and Friedman, H.
2010. The regulation of MADS-box gene expression during ripening
of banana and their regulatory interaction with ethylene. Journal of
Experimental Botany 61, 1523–1535.
Enfissi, E.M., Barneche, F., Ahmed, I., Lichtle, C., Gerrish, C., McQuinn, R.P.,
Giovannoni, J.J., Lopez-Juez, P., Bowler, C., Bramley, P.M., and Fraser, P.D.
2010. Integrative transcript and metabolite analysis of nutritionally enhanced
DEETIOLATED1 downregulated tomato fruit. Plant Cell 22, 1190–1215.
Eriksson, E.M., Bovy, A., Manning, K., Harrison, L., Andrews, J., De Silva, J.,
Tucker, G.A., and Seymour, G.B. 2004. Effects of the colourless nonripen-
ing (Cnr) mutation on gene expression and cell wall biochemistry during
tomato fruit development and ripening. Plant Physiology 136, 4184–4197.

327
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Eriksson, O., Friis, E.M., and Lofgren, P. 2000. Seed size, fruit size, and dis-
persal systems in angiosperms from the early cretaceous to the late ter-
tiary. American Naturalist 156, 47–58.
Espley, R.V., Brendolise, C., Chagne, D., Kutty-Amma, S., Green, S., Volz, R.,
Putterill, J., Schouten, H.J., Gardiner, S.E., Hellens, R.P., and Allan, A.C.
2009. Multiple repeats of a promoter segment causes transcription fac-
tor autoregulation in red apples. Plant Cell 21, 168–183.
Espley, R.V., Hellens, R.P., Putterill, J., Stevenson, D.E., Kutty-Amma, S., and
Allan, A.C. 2007. Red colouration in apple fruit is due to the activity of
the MYB transcription factor, MdMYB10. Plant Journal 49, 414–427.
Ezura, H., and Owino, W.O. 2008. Melon, an alternative model plant for eluci-
dating fruit ripening. Plant Science 175, 121–129.
Fait, A., Hanhineva, K., Beleggia, R., Dai, N., Rogachev, I., Nikiforova, V.J.,
Fernie, A.R., and Aharoni, A. 2008. Reconfiguration of the achene and
receptacle metabolic networks during strawberry fruit development.
Plant Physiology 148, 730–750.
Faria, M.V., Maluf, W.R., Vilela de Resende, J.T., Andrade-Junior, V.C.,
do Nascimento, I.R., Gandolfi Benites, F.R., de Menezes, C.B., and
Azevedo, S.M. 2006. Rin, norA, ogc and hp mutants in tomatoes with
different genetic backgrounds. Pesquisa Agropecuaria Brasileira 41,
793–800.
Fedorof f, N.V. 2002. Cross-talk in abscisic acid signaling. Science
STKE 140.
Finamore, A., Roselli, M., Britti, S., Monastra, G., Ambra, R., Turrini, A.,
and Mengheri, E. 2008. Intestinal and peripheral immune response
to MON810 maize ingestion in weaning and old mice. Journal of
Agricultural and Food Chemistry 56, 11533–11539.
Finkelstein, R.R., and Rock, C.D. 2002. Abscisic acid biosynthesis and
response. Arabidopsis Book 1:e0058.
Flores, F., El Yahyaoui, F., de Billerbeck, G., Romojaro, F., Latche, A.,
Bouzayen, M., Pech, J.C., and Ambid, C. 2002. Role of ethylene in the
biosynthetic pathway of aliphatic ester aroma volatiles in Charentais
cantaloupe melons. Journal of Experimental Botany 53, 201–206.
Fraser, P.D., and Bramley, P.M. 2004. The biosynthesis and nutritional uses of
carotenoids. Progress in Lipid Research 43, 228–265.
Fraser, P.D., Enfissi, E.M., and Bramley, P.M. 2009. Genetic engineering of
carotenoid formation in tomato fruit and the potential application of sys-
tems and synthetic biology approaches. Archives of Biochemistry and
Biophysics 483, 196–204.
Fraser, P.D., Enfissi, E.M, Halket, J.M., Truesdale, M.R., Yu, D., Gerrish, C.,
and Bramley, P.M. 2007. Manipulation of phytoene levels in tomato fruit:
Effects on isoprenoids, plastids, and intermediary metabolism. Plant
Cell 19, 3194–3211.
Fujisawa, M., Nakano, T., and Ito, Y. 2011. Identification of potential target
genes for the tomato fruit-ripening regulator RIN by chromatin immu-
noprecipitation. BMC Plant Biology 11, 26.

328
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Fujisawa, M., Nakano, T., Shima, Y., and Ito, Y. 2013. A large-scale identifi-
cation of direct targets of the tomato MADS box transcription factor
RIPENING INHIBITOR reveals the regulation of fruit ripening. Plant
Cell 25, 371–386.
Galpaz, N., Wang, Q., Menda, N., Zamir, D., and Hirschberg, J. 2008.
Abscisic acid deficiency in the tomato mutant high-pigment 3 leading
to increased plastid number and higher fruit lycopene content. Plant
Journal 53, 717–730.
Giménez, E., Pineda, B., Capel, J., Anton, M.T., Atares, A., Perez- Martin, F.,
Garcia-Sogo, B., Angosto, G., Moreno, V., and Lozano, R. 2010.
Functional analysis of the Arlequin mutant corroborates the essential
role of the Arlequin/TAGL1 gene during reproductive development of
tomato. PLoS ONE 5, e14427.
Giorio, G., Stigliani, A.L., and D’Ambrosio, C. 2007. Agronomic performance
and transcriptional analysis of carotenoid biosynthesis in fruits of
transgenic HighCaro and control tomato lines under field conditions.
Transgenic Research 16, 15–28.
Giovannoni, J. 2001. Molecular biology of fruit maturation and ripening.
Annual Review of Plant Physiology and Plant Molecular Biology 52,
725–749.
Giovannoni, J.J. 2004. Genetic regulation of fruit development and ripening.
Plant Cell 16, 170–180.
Giovannoni, J.J. 2007. Fruit ripening mutants yield insights into ripening con-
trol. Current Opinion in Plant Biology 10, 283–289.
Giovannoni, J.J., Noensie, E.N., Ruezinsky, D.M., Lu, X., Tracy, S.L., Ganal,
M.W., Martin, G.B., Pillen, K., Alpert, K., and Tanksley, S.D. 1995.
Molecular genetic analysis of the ripening inhibitor and non-ripening
loci of tomato: A first step in genetic map based cloning of fruit ripening
genes. Molecular Genetics and Genomics 248, 195–206.
Girin, T., Stephenson, P., Goldsack, C.M.P., Kempin, S.A., Perez, A., Pires,
N., Sparrow, P.A., Wood, T.A., Yanofsky, M.F., and Osterggard, L. 2010.
Brassicaceae INDEHISCENT genes specify valve margin cell fate and
repress replum formation. Plant Journal 63, 329–338.
Gorguet, B., Eggink, P.M., Ocana, J., Tiwari, A., Schipper, D., Finkers, R.,
Visser, R.G., and van Heusden, A.W. 2008. Mapping and characteriza-
tion of novel parthenocarpy QTLs in tomato. Theoretical and Applied
Genetics 116, 755–767.
Goulao, L.F., and Oliveira, C.M. 2007. Cell wall modifications during fruit rip-
ening: when a fruit is not the fruit. Trends in Food Science and Technology
19, 4–25.
Grierson, D. 2013. Ethylene and the control of fruit ripening. In Molecular
Biology and Biochemistry of Fruit Ripening, ed. G.B. Seymour, J.J.
Giovannoni, G.A. Tucker, M. Poole. Boston: Wiley, 43–74.
Hileman, L., Sundstrom, J., Litt, A., Chen, M., Shumba, T., and Irish, V. 2006.
Molecular and phylogenetic analyses of the MADS-box gene family in
tomato. Molecular Biology and Evolution 23, 2245–2258.

329
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Himmelbach, A., Yang, Y., and Grill, E. 2003. Relay and control of abscisic
acid signaling. Current Opinion in Plant Biology 6, 470–479.
Hirayama, T., and Shinozaki, K. 2007. Perception and transduction of abscisic
acid signals: Keys to the function of the versatile plant hormone ABA.
Trends in Plant Science 12, 343–351.
Hospital, F. 2009. Challenges for effective marker-assisted selection in plants.
Genetica 136, 303–310.
Hou, D.X., Fujii, M., Terahara, N., and Yoshimoto, M. 2004. Molecular mecha-
nisms behind the chemopreventive effects of anthocyanidins. Journal of
Biomedicine and Biotechnology 5, 321–325.
Hovav, R., Chehanovsky, N., Moy, M., Jetter, R., and Schaffer, A.A. 2007. The
identification of a gene (Cwp1), silenced during Solanum evolution,
which causes cuticle microfissuring and dehydration when expressed
in tomato fruit. Plant Journal 52, 627–639.
Iannetta, P.P.M., Laarhovenb, L.J., Medina-Escobar, N., James, E.K.,
McManuse, M.T., Davies, H.V., and Harren, F.J.M. 2006. Ethylene and
carbon dioxide production by developing strawberries show a correla-
tive pattern that is indicative of ripening climacteric fruit. Physiologia
Plantarum 127, 247–259.
Ireland, H.S., Yao, J.L., Tomes, S., Sutherland, P.W., Nieuwenhuizen, N.,
Gunaseelan, K., Winz, R.A., David, K.M., and Schaffer, R.J. 2013. Apple
SEPALLATA1/2-like genes control fruit flesh development and ripen-
ing. Plant Journal 73, 1044–1056.
Isaacson, T., Kosma, D.K., Matas, A.J., Buda, G.J., He, Y., Yu, B., Pravitasari,
A., Batteas, J.D., Stark, R.E., Jenks, M.A., and Rose, J.K.C. 2009. Cutin
deficiency in the tomato fruit cuticle consistently affects resistance to
microbial infection and biomechanical properties, but not transpira-
tional water loss. Plant Journal 60, 363–377.
Jaakola, L., Poole, M., Jones, M.O., Kamarainen-Karppinen, T., Koskimaki,
J.J., Hohtola, A., Haggman, H., Fraser, P.D., Manning, K., King, G.J.,
Thomson, H., and Seymour, G.B. 2010. A SQUAMOSA MADS box
gene involved in the regulation of anthocyanin accumulation in bilberry
fruits. Plant Physiology 153, 1619–1629.
Jaillon, O., Aury, J.M., Noel, B., Policriti, A., Clepet, C., Casagrande, A.,
Choisne, N., Aubourg, S., Vitulo, N., Jubin, C., Vezzi, A., Legeai, F.,
Hugueney, P., Dasilva, C., Horner, D., Mica, E., Jublot, D., Poulain,
J., Bruyere, C., Billault, A., Segurens, B., Gouyvenoux, M., Ugarte,
E., Cattonaro, F., Anthouard, V., Vico, V., Del Fabbro, C., Alaux,
M., Di Gaspero, G., Dumas, V., Felice, N., Paillard, S., Juman, I.,
Moroldo, M., Scalabrin, S., Canaguier, A., Le Clainche, I., Malacrida,
G., Durand, E., Pesole, G., Laucou, V., Chatelet, P., Merdinoglu, D.,
Delledonne, M., Pezzotti, M., Lecharny, A., Scarpelli, C., Artiguenave,
F., Pe, M.E., Valle,  G., Morgante, M., Caboche, M., Adam-Blondon,
A.F., Weissenbach, J., Quetier, F., and Wincker, P. 2007. The grapevine
genome sequence suggests ancestral hexaploidization in major angio-
sperm phyla. Nature 449, 463–465.

330
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Jia, H.F., Chai, Y.M., Li, C.L., Lu, D., Luo, J.J., Qin, L., and Shen, Y. 2011.
Abscisic acid plays an important role in the regulation of strawberry
fruit ripening. Plant Physiology 157, 188–199.
Jiang, Y., Joyce, D.C., and Macnish, A.J. 2000. Effect of abscisic acid on
banana fruit ripening in relation to the role of ethylene. Journal of Plant
Growth Regulation 19, 106–111.
Jiang, Y.M., and Joyce, D.C. 2003. ABA effects on ethylene production, PAL
activity, anthocyanin and phenolic contents of strawberry fruit. Plant
Growth Regulation 39, 171–174.
Johnston, J.W., Gunaseelan, K., Pidakala, P., Wang, M., and Schaffer, R.J. 2009.
Co-ordination of early and late ripening events in apples is regulated
through differential sensitivities to ethylene. Journal of Experimental
Botany 60, 2689–2699.
Jones, B., Frasse, P., Olmos, E., Zegzouti, H., Li, Z.G., Latche, A., Pech, J.C.,
and Bouzayen, M. 2002. Down-regulation of DR12, an auxin-response-
factor homolog, in the tomato results in a pleiotropic phenotype includ-
ing dark green and blotchy ripening fruit. Plant Journal 32, 603–613.
Kanno, Y., Jikumaru, Y., Hanada, A., Nambara, E., Abrams, S.R., Kamiya,
Y., and Seo, M. 2010. Comprehensive hormone profiling in develop-
ing Arabidopsis seeds: Examination of the site of ABA biosynthesis,
ABA transport and hormone interactions. Plant and Cell Physiology 51,
1988–2001.
Karlova, R., Rosin, F.M., Busscher-Lange, J., Parapunova, V., Do, P.T., Fernie,
A.R., Fraser, P.D., Baxter, C., Angenent, G.C., and de Maagd, R.A. 2011.
Transcriptome and metabolite profiling show that APETALA2a is a
major regulator of tomato fruit ripening. Plant Cell 23, 923–941.
Klee, H.J., and Giovannoni, J.J. 2011. Genetics and control of tomato fruit rip-
ening and quality attributes. Annual Review of Genetics 45, 41–59.
Klie, S., Krueger, S., Krall, L., Giavalisco, P., Flügge, U.I., Willmitzer, L., and
Steinhouser, D. 2011. Analysis of the compartmentalized metabolome—
A validation of the non-aqueous fractionation technique. Frontiers in
Plant Science 2, 55.
Knapp, S. 2002. Tobacco to tomatoes: A phylogenetic perspective on fruit diver-
sity in the Solanaceae. Journal of Experimental Botany 53, 2001–2022.
Kobashi, K., Gemma, H., and Iwahori, S. 1999. Sugar accumulation in peach
fruit as affected by abscisic acid treatment in relation to some sugar
metabolizing enzymes. Journal of the Japanese Society for Horticultural
Science 68, 465–470.
Kou, X., Watkins, C.B., and Gan, S.S. 2012. Arabidopsis AtNAP regulates fruit
senescence. Journal of Experimental Botany 63, 6139–6147.
Kumar, R., Sharma, M.K., Kapoor, S., Tyagi, A.K., and Sharma, A.K. 2012.
Transcriptome analysis of rin mutant fruit and in silico analysis of pro-
moters of differentially regulated genes provides insight into LeMADS-
RIN-regulated ethylene-dependent as well as ethylene-independent
aspects of ripening in tomato. Molecular Genetics and Genomics 287,
189–203.

331
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lanahan, M.B., Yen, H.C., Giovannoni, J.J., and Klee, H.J. 1994. The never ripe
mutation blocks ethylene perception in tomato. Plant Cell 6, 521–530.
Larsson, S.C., Bergkvist, L., and Wolk, A. 2009. Glycemic load, glycemic
index and breast cancer risk in a prospective cohort of Swedish women.
International Journal of Cancer 125, 153–157.
Law, J.A., and Jacobsen, S.E. 2010. Establishing, maintaining and modifying
DNA methylation patterns in plants and animals. Nature 11, 204–220.
Lee, J.M., Joung, J.G., McQuinn, R., Chung, M.Y., Fei, Z., Tieman, D., Klee,
H., and Giovannoni, J. 2012. Combined transcriptome, genetic diver-
sity and metabolite profiling in tomato fruit reveals that the ethylene
response factor SlERF6 plays an important role in ripening and carot-
enoid accumulation. Plant Journal 70, 191–204.
Leide, J., Hildebrandt, U., Reussing, K., Riederer, M., and Vogg, G. 2007. The
developmental pattern of tomato fruit wax accumulation and its impact on
cuticular transpiration barrier properties: Effects of a deficiency in a beta-
ketoacyl-coenzyme A synthase (LeCER6). Plant Physiology 144, 1667–1679.
Leung, J., and Giraudat, J. 1998. Abscisic acid signal transduction. Annual
Review of Plant Physiology and Plant Molecular Biology 49, 199–222.
Lin, Z., Arciga-Reyes, L., Zhong, S., Alexander, L., Hackett, R., Wilson, I., and
Grierson, D. 2008. SlTPR1, a tomato tetratricopeptide repeat protein,
interacts with the ethylene receptors NR and LeETR1, modulating eth-
ylene and auxin responses and development. Journal of Experimental
Botany 59, 4271–4287.
Lippman, Z.B., Semel, Y., and Zamir, D. 2007. An integrated view of quan-
titative trait variation using tomato interspecific introgression lines.
Current Opinion in Genetics Development 17, 545–552.
Liu, K.D., Kang, B.C., Jiang, H., Moore, S.L., Li, H.X., Watkins, C.B., Setter,
T.L., and Jahn, M.M. 2005. AGH3-like gene, CcGH3, isolated from
Capsicum chinense L. fruit is regulated by auxin and ethylene. Plant
Molecular Biology 58, 447–464.
Liu, Y., Roof, S., Ye, Z., Barry, C., van Tuinen, A., Vrebalov, J., Bowler, C.,
and Giovannoni, J. 2004. Manipulation of light signal transduction as a
means of modifying fruit nutritional quality in tomato. Proceedings of the
National Academy of Sciences USA 101, 9897–9902.
Lohani, S., Trivedi, P.K., and Nath, P. 2004. Changes in activities of cell wall
hydrolases during ethylene induced ripening in banana: effect of 1-MCP,
ABA and IAA. Postharvest Biology and Technology 31, 119–126.
Luo, J., Butelli, E., Hill, L., Parr, A., Niggeweg, R., Bailey, P., Weisshaar, B.,
and Martin, C. 2008. AtMYB12 regulates caffeoyl quinic acid and flavo-
nol synthesis in tomato: Expression in fruit results in very high levels of
both types of polyphenol. Plant J 56, 316–326.
Mann, J. 2004. Free sugars and human health: Sufficient evidence for action?
Lancet 363, 1068–1070.

332
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Manning, K., Tor, M., Poole, M., Hong, Y., Thompson, A.J., King, G.J.,
Giovannoni, J.J., and Seymour, G.B. 2006. A naturally occurring epigen-
etic mutation in a gene encoding an SPB-box transcription factor inhib-
its tomato fruit ripening. Nature Genetics 38, 949–952.
Martel, C., Vrebalov, J., Tafelmeyer, P., and Giovannoni, J.J. 2011. The tomato
MADS-box transcription factor RIPENING INHIBITOR interacts with
promoters involved in numerous ripening processes in a COLORLESS
NONRIPENING-dependent manner. Plant Physiology 157, 1568–1579.
Martin, C., Zhang, Y., Tonelli, C., and Petroni, K. 2013. Plants, diet, and
health. Annual Review of Plant Biology 64, 19–46.
Martinez, G.A., Chaves, A.R., and Anon, M.C. 1996. Effect of exogenous
application of gibberelic acid on color change and phenylalanine ammo-
nia lyase, chlorophyllase, and peroxidase activities during ripening of
strawberry fruit (Fragaria × ananassa Duch). Journal of Plant Growth
Regulation 15, 139–146.
Meli, V.S., Ghosh, S., Prabha, T.N., Chakraborty, N., Chakraborty, S., and
Datta, A. 2010. Enhancement of fruit shelf life by suppressing N-glycan
processing enzymes. Proceedings of National Academy of Sciences USA
107, 2413–2418.
Messeguer, R., Ganal, M.W., Steffens, J.C., and Tanksley, S.D. 1991.
Characterization of the level, target sites and inheritance of cyto-
sine methylation in tomato nuclear DNA. Plant Molecular Biology 16,
753–770.
Moreno, E., Obando, J.M., Dos-Santos, N., Fernandez-Trujillo, J.P., Monforte,
A.J., and Garcia-Mas, J. 2008. Candidate genes and QTLs for fruit rip-
ening and softening in melon. Theoretical and Applied Genetics 116,
589–602.
Morris, J., Hawthorne, K.M., Hotze, T., Abrams, S.A., and Hirschi, K.D. 2008.
Nutritional impact of elevated calcium transport activity in carrots.
Proceedings of National Academy of Sciences USA, 105, 1431–1435.
Mustilli, A.C., Fenzi, F., Ciliento, R., Alfano, F., and Bowler, C. 1999. Phenotype
of the tomato high pigment-2 mutant is caused by a mutation in the
tomato homolog of DEETIOLATED1. Plant Cell 11, 145–157.
Obando, J., Fernandez-Trujillo, J.P., Martinez, J.A., Alarcon, A.L., Eduardo,
I., Arus, P., and Monforte, A.J. 2008. Identification of melon fruit quality
quantitative trait loci using near-isogenic lines. Journal of the American
Society for Horticultural Science 133, 139–151.
Osorio, S., Alba, R., Damasceno, C.M., Lopez-Casado, G., Lohse, M.,
Zanor, M.I., Tohge, T., Usadel, B., Rose, J.K., Fei, Z., Giovannoni, J.J.,
and Fernie, A.R. 2011. Systems biology of tomato fruit development:
Combined transcript, protein, and metabolite analysis of tomato tran-
scription factor (nor, rin) and ethylene receptor (Nr) mutants reveals
novel regulatory interactions. Plant Physiology 157, 405–425.

333
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Osorio, S., Alba, R., Nikoloski, Z., Kochevenko, A., Fernie, A.R., and
Giovannoni, J.J. 2012. Integrative comparative analyses of transcript
and metabolite profiles from pepper and tomato ripening and develop-
ment stages uncover species-specific patterns of network regulatory
­behavior. Plant Physiology 159, 1713–1729.
Pan, Q.H., Li, M.J., Peng, C.C., Zhang, N., Zou, X., Zou, K.Q., Wang, X.L.,
Yu, X.C., Wang, X.F., and Zhang, D.P. 2005. Abscisic acid activates acid
­invertases in developing grape berry. Physiologia Plantarum 125, 157–170.
Park, S., Cheng, N.H., Pittman, J.K., Yoo, K.S., Park, J., Smith, R.H., and
Hirschi, K.D. 2005. Increased calcium levels and prolonged shelf life in
tomatoes expressing Arabidopsis H+. Plant Physiology 139, 1194–1206.
Patterson, G.I., Thorpe, C.J., and Chandler, V.L. 1993. Paramutation, an allelic
interaction, is associated with a stable and heritable reduction of tran-
scription of the maize regulatory gene. Genetics 135, 881–894.
Paul, V., Pandey, R., and Srivastava, G.C. 2012. The fading distinctions
between classical patterns of ripening in climacteric and non-climacteric
fruit and the ubiquity of ethylene—An overview. Journal of Food Science
and Technology 49, 1–21.
Perkins-Veazie, P.M. 1995. Growth and ripening of strawberry fruit.
Horticultural Reviews 17, 267–297.
Perkins-Veazie, P.M., Huber, D.J., and Brecht, J.K. 1996. In vitro growth and
ripening of strawberry fruit in presence of ACC, STS or propylene.
Annals of Applied Biology 128, 105–116.
Phillips, P.C. 2008. Epistasis—The essential role of gene interactions in the
structure and evolution of genetic systems. Nature Reviews Genetics 9,
855–867.
Pilcher, R.L., Moxon, S., Pakseresht, N., Moulton, V., Manning, K.,
Seymour, G., and Dalmay, T. 2007. Identification of novel small RNAs in
tomato (Solanum lycopersicum). Planta 226, 709–717.
Powell, A.L., Nguyen, C.V., Hill, T., Cheng, K.L., Figueroa-Balderas, R.,
Aktas,  H., Ashrafi, H., Pons, C., Fernandez-Munoz, R., Vicente, A.,
Lopez-Baltazar, J., Barry, C.S., Liu, Y., Chetelat, R., Granell, A., Van
Deynze, A., Giovannoni, J.J., and Bennett, A.B. 2012. Uniform ripen-
ing encodes a Golden 2-like transcription factor regulating tomato fruit
chloroplast development. Science 336, 1711–1715.
Powell, A.L.T., Kalamaki, M.S., Kurien, P.A., Gurrieri, S., and Bennett, A.B.
2003. Simultaneous transgenic suppression of LePG and LeExp1 influ-
ences fruit texture and juice viscosity in a fresh market tomato variety.
Journal of Agricultural and Food Chemistry 51, 7450–7455.
Richings, E.W., Cripps, R.F., and Cowan, A.K. 2000. Factors affecting ‘Hass’ avo-
cado fruit size: Carbohydrate, abscisic acid and isoprenoid metabolism in
normal and phenotypically small fruit. Physiologia Plantarum 109, 81–89.
Rodrigo, M.J., Marcos, J.F., Alferez, F., Mallent, M.D., and Zacarias, L. 2003.
Characterization of Pinalate, a novel Citrussinensis mutant with a fruit-
specific alteration that results in yellow pigmentation and decreased
ABA content. Journal of Experimental Botany 54, 727–738.

334
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Saladie, M., Matas, A.J., Isaacson, T., Jenks, M.A., Goodwin, S.M., Niklas, K.J.,
Xiaolin, R., Labavitch, J.M., Shackel, K.A., Fernie, A.R., Lytovchenko,
A., O’Neill, M.A., Watkins, C.B., and Rose, J.K. 2007. A reevaluation of
the key factors that influence tomato fruit softening and integrity. Plant
Physiology 144, 1012–1028.
Schaffer, R.J., Friel, E.N., Souleyre, E.J.F., Bolitho, K., Ledger, K.T.S.,
Bowen, J.H., Ma, J.H., Nain, B., Cohen, D., Gleave, A.P., Crowhurst,
R.N., Janssen, B.J., Yao, J.L., and Newcomb, R.D. 2007. A genomics
approach reveals that aroma production in apple is controlled by ethyl-
ene predominantly at the final step in each biosynthetic pathway. Plant
Physiology 144, 1899–1912.
Schijlen, E.G., de Vos, C.H., Martens, S., Jonker, H.H., Rosin, F.M., Molthoff,
J.W., Tikunov, Y.M., Angenent, G.C., van Tunen, A.J., and Bovy, A.G.
2007. RNA interference silencing of chalcone synthase, the first step
in the flavonoid biosynthesis pathway, leads to parthenocarpic tomato
fruits. Plant Physiology 144, 1520–1530.
Setha, S. 2012. Roles of abscisic acid in fruit ripening. Walailak Journal of
Science and Technology 9, 297–308.
Seymour, G., Poole, M., Manning, K., and King, G.J. 2008. Genetics and epi-
genetics of fruit development and ripening. Current Opinion in Plant
Biology 11, 58–63.
Seymour, G.B., Ryder, C.D., Cevik, V., Hammond, J.P., Popovich, A., King,
G.J., Vrebalov, J., Giovannoni, J.J., and Manning, K. 2011. A SEPALLATA
gene is involved in the development and ripening of strawberry
(Fragaria × ananassa Duch.) fruit, a non-climacteric tissue. Journal of
Experimental Botany 62, 1179–1188.
Shulaev, V., Sargent, D.J., Crowhurst, R.N., Mockler, T.C., Folkerts, O.,
Delcher, A.L. et al., 2011. The genome of woodland strawberry (Fragaria
vesca). Nature Genetics 43, 109–116.
Stephenson, P., Baker, D., Girin, T., Perez, A., Amoah, S., King, G.J., and
Ostergaard, L. 2010. A rich TILLING resource for studying gene func-
tion in Brassica rapa. BMC Plant Biology 10, 62.
Stevens, R., Buret, M., Duffe, P., Garchery, C., Baldet, P., Rothan, C., and
Causse, M. 2007. Candidate genes and quantitative trait loci affecting
fruit ascorbic acid content in three tomato populations. Plant Physiology
143, 1943–1953.
Sun, L., Sun, Y., Zhang, M., Wang, L., Ren, J., Cui, M., Wang, Y., Ji, K., Li, P.,
Li, Q., Chen, P., Dai, S., Duan, C., Wu, Y., and Leng, P. 2012. Suppression
of 9-cis-epoxycarotenoid dioxygenase, which encodes a key enzyme
in abscisic acid biosynthesis, alters fruit texture in transgenic tomato.
Plant Physiology 158, 283–298.
Sun, Y., Chen, P., Duan, C., Tao, P., Wang, Y., Ji, K., Hu, Y., Li, Q., Dai, S.,
Wu, Y., Luo, H., Sun, L., and Leng, P. 2012. Transcriptional regulation of
genes encoding key enzymes of abscisic acid metabolism during melon
(Cucumis melo L.) fruit development and ripening. Journal of Plant
Growth Regulation 32, 233–244.

335
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Symons, G.M., Davies, C., Shavrukov, Y., Dry, I.B., Reid, J.B., and Thomas,
M.R. 2006. Grapes on steroids. Brassinosteroids are involved in grape
berry ripening. Plant Physiology 140, 150–158.
Thom, E. 2007. The effect of chlorogenic acid enriched coffee on glucose
absorption in healthy volunteers and its effect on body mass when used
long-term in overweight and obese people. Journal of International
Medical Research 35, 900–908.
Thompson, A.J., Tor, M., Barry, C.S., Vrebalov, J., Orfila, C., Jarvis, M.C.,
Giovannoni, J.J., Grierson, D., and Seymour, G.B. 1999. Molecular and
genetic characterization of a novel pleiotropic tomato ripening mutant.
Plant Physiology 120, 383–390.
Tomato Genome Consortium. 2012. The tomato genome sequence provides
insights into fleshy fruit evolution. Nature 485, 635–641.
Trainotti, L., Spinello, R., Piovan, A., Spolaore, S., and Casadoro, G. 2001. Beta-
galactosidases with a lectin-like domain are expressed in strawberry.
J. Exp. Bot. 52, 1635–1645.
Trainotti, L., Tadiello, A., and Casadoro, G. 2007. The involvement of auxin
in the ripening of climacteric fruits comes of age: The hormone plays
a role of its own and has an intense interplay with ethylene in ripening
peaches. Journal of Experimental Botany 58, 3299–3308.
Tsuda, T., Horio, F., Uchida, K., Aoki, H., and Osawa, T. 2003. Dietary cyani-
din 3-O-β-D-glucoside-rich purple corn color prevents obesity and ame-
liorates hyperglycemia in mice. Journal of Nutrition 133, 2125–2130.
Vaughn, M.W., Tanurdzic, M., Lippman, Z., Jiang, H., Carrasquillo, R.,
Rabinowicz, P.D., Dedhia, N., McCombi, W.R., Agier, N., Bulski, A.,
Colot, V., Doerge, R.W., and Martienssen, R.A. 2007. Epigenetic natural
variation in Arabidopsis thaliana. PLoS Biology 5, e174.
Venneria, E., Fanasca, S., Monastra, G., Finotti, E., Ambra, R., Azzini, E.,
Durazzo, A., Foddai, M.S., and Maiani, G. 2008. Assessment of the nutri-
tional values of genetically modified wheat, corn, and tomato crops.
Journal of Agricultural and Food Chemistry 56, 9206–9214.
Vicente, A.R., Saladié, M., Rose, J.K.C., and Labavitch, J.M. 2007. The linkage
between cell wall metabolism and fruit softening: Looking to the future.
Journal of the Science of Food and Agriculture 87, 1435–1448.
Vrebalov, J., Pan, I.L., Arroyo, A.J., McQuinn, R., Chung, M., Poole, M.,
Rose, J., Seymour, G., Grandillo, S., Giovannoni, J., and Irish, V.F.
2009. Fleshy fruit expansion and ripening are regulated by the tomato
SHATTERPROOF gene, TAGL1. Plant Cell 21, 3041–3062.
Vrebalov, J., Ruezinsky, D., Padmanabhan, V., White, R., Medrano, D., Drake,
R., Schuch, W., and Giovannoni, J. 2002. A MADS-box gene necessary
for fruit ripening at the tomato ripening-inhibitor (rin) locus. Science
296, 343–346.
Wada, H., Shackel, K.A., and Matthews, M.A. 2008. Fruit ripening in Vitis
vinifera: Apoplastic solute accumulation accounts for pre-veraison
­turgor loss in berries. Planta 227, 1351–1361.

336
Mo lecul ar Regul ati o n o f Fru it Ripen in g

Wang, H., Schauer, N., Usadel, B., Frasse, P., Zouine, M., Hernould, M.,
Latche, A., Pech, J.C., Farnie, A.R., and Bouzayen, M. 2009. Regulatory
features underlying pollination-dependent and -independent tomato
fruit set revealed by transcript and primary metabolite profiling. Plant
Cell 21, 1428–1452.
Willett, W.C. 2010. Fruits, vegetables, and cancer prevention: Turmoil in the
produce section. Journal of the National Cancer Institute 102, 510–511.
Yamaki, S., and Asakura, T. 1991. Stimulation of the uptake of sorbitol into
vacuoles from apple fruit flesh by abscisic acid and into protoplasts by
indoleacetic acid. Plant Cell Physiol. 32, 315–318.
Ytterberg, A.J., Peltier, J.B., and van Wijk, K.J. 2006. Protein profiling of
plastoglobules in chloroplasts and chromoplasts. A surprising site for
differential accumulation of metabolic enzymes. Plant Physiology 140,
984–997.
Zahn, L.M., Komg, H., Leebens-Mack, J.H., Kim, S., Soltis, P.S., Landherr, L.L.,
Soltis, D.E., dePamphilis, C.W., and Ma, H. 2005. The evolution of the
SEPALLATA subfamily of MADS-box genes. Genetics 169, 2209–2223.
Zhang, B., Chen, K., Bowen, J., Allan, A., Espley, R., Karunairetnam, S., and
Ferguson, I. 2006. Differential expression within the LOX gene family in
ripening kiwifruit. Journal of Experimental Botany 57, 3825–3836.
Zhang, M., Leng, P., Zhang, G., and Li, X. 2009a. Cloning and functional analy-
sis of 9-cis-epoxycarotenoid dioxygenase (NCED) genes encoding a key
enzyme during abscisic acid biosynthesis from peach and grapefruits.
Journal of Plant Physiology 166, 1241–1252.
Zhang, M., Yuan, B., and Leng, P. 2009b. The role of ABA in triggering ethyl-
ene biosynthesis and ripening of tomato fruit. Journal of Experimental
Botany 60, 1579–1588.
Zhang, X., Richards, E.J., and Borevitz, J.O. 2007. Genetic and epigenetic dis-
section of cis regulatory variation. Current Opinion in Plant Biology 10,
142–148.
Zhi, Q., Wang, S., Chai, M., Zhang, F., Li, Q., Li, S., and Sun, M. 2007.
Transgenic mini-tomato and protection against alcohol-induced gastric
injury. Journal of Genetics and Genomics 34, 756–763.
Zhong, S., Fei, Z., Chen, Y.R., Zheng, Y., Huang, M., Vrebalov, J., McQuinn, R.,
Gapper, N., Liu, B., Xiang, J., Shao, Y., and Giovannoni, J.J. 2013. Single-
base resolution methylomes of tomato fruit development reveal epig-
enome modifications associated with ripening. Nature Biotechnology 31,
154–159.
Zilberman, D., and Henikoff, S. 2007. Genome-wide analysis of DNA methyla-
tion patterns. Development 134, 3959–3965.

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Chapter 11

Advances in Ethylene
Signal Transduction in
Fruits and Vegetables
Willis O. Owino1 and Jane Ambuko2
1Jomo Kenyatta University of Agriculture and
Technology, Nairobi, Kenya
2University of Nairobi, Nairobi, Kenya

Abstract 340
11.1 Introduction 340
11.2 Ethylene Physiology in Climacteric and
Nonclimacteric Fruits 341
11.3 Ethylene Signaling Components in Fruits 342
11.4 Analyses of Ethylene Signal Components in
Other Fruit Species 344
11.4.1  Climacteric Fruits 344
11.4.2  Nonclimacteric Fruits 348
11.5 Transcriptional Regulators of Fruit Ripening 349
11.6 Conclusions and Future Perspectives 352
References 353

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Abstract
Fruits and vegetables are a significant part of a healthy diet, as they are a
source of essential vitamins, minerals, fiber, and other health-promoting
compounds. The major challenges in improvement of these commodities is
the betterment of their health-promoting attributes while improving qual-
ity, postharvest shelf life, marketability, processing qualities, and consumer
appeal. To address these challenges, researchers have pursued strategies
of understanding key control points in global development regulation or
within specific processes in fruits and vegetables. This is informed by the
substantial insights into the mechanistic basis of ethylene biosynthesis,
perception, and signaling, and the identity of key regulators of ripening that
operate upstream of, or in concert with, regulatory pathways that mediate
the plant hormone ethylene. In this chapter, we highlight the contributions
made in our understanding of ethylene signaling, ethylene physiology,
and ethylene signaling components in both climacteric and nonclimac-
teric fruits. Tomato is the model system of choice for studying ethylene
signaling in fleshy fruit due to its commercial importance, a rich source of
genetic and biochemical information, relatively small genome, relative ease
of genetic transformation, and availability of developmental mutants that
are ripening impaired. Meanwhile, we look at how the translation of infor-
mation from tomato to other fleshy-fruited species indicates that ethylene
signaling and regulatory networks are conserved across a wide spectrum
of fruits and vegetables. We also discuss the ethylene additional regulatory
systems that coordinate the ethylene production. We finally summarize the
future perspectives of research in the ethylene signaling pathway.

11.1 Introduction
The phytohormone ethylene is a simple gaseous alkene that is a potent
modulator of proper plant development, growth, and survival (Bleecker
and Kende, 2000). The crucial role that ethylene plays in regulating mul-
tiple physiological and developmental processes in plants, such as leaf
and flower senescence, ripening, organ abscission, and growth transition,
and its involvement in the reactions of plants to abiotic and biotic stresses
are well documented (Chang and Bleecker, 2004; Seymour et al., 2013a).
Ethylene is physiologically active in trace amounts, and its effects are of
great commercial importance. To understand the roles of ethylene in plant
functions, it is important to know how this gaseous hormone is synthesized,
how its production is regulated, and how the signal is transduced.
Major breakthroughs in understanding how the ethylene signal is per-
ceived and transduced to mediate phenotypic responses have been made

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through genetic studies in the model plant Arabidopsis, resulting in the

­identification of critical components of this signaling pathway (Kendrick and
Chang, 2008; Stepanova and Alonso, 2009; Zhao and Guo, 2011; Merchante
et al., 2013). The Arabidopsis has been and is still the ideal model plant for
many genetic and molecular analyses, and without it, it would have been
impractical to assemble such a comprehensive understanding of the indi-
vidual components of the perception of ethylene (Klee, 2004).
However, fleshy fruits are dissimilar to Arabidopsis in the sense that
they undergo a ripening process in which the biochemistry, physiology, and
structure of the organ are developmentally altered to influence the appear-
ance, texture, flavor, and aroma in ways designed to attract seed-dispersing
organisms (Giovannoni, 2004). The specific biochemical programs culmi-
nating in ripening differ from one fleshy fruit to another. The importance of
ethylene in regulating traits of agronomic importance, particularly fruit rip-
ening, has driven research on the identification and functional characteriza-
tion of components of the ethylene signaling pathway in crop species. The
tomato has been at the forefront of the comparative analysis. Comparative
research between the regulation of ethylene signaling in Arabidopsis and
crops of agronomic importance such as tomato indicates significant dif-
ferences in terms of gene family composition and expression (Cara and
Giovannoni, 2008). However, the fundamental genetic building blocks of
ethylene signal transduction systems are very similar. Extending the shelf
life by delaying the biosynthesis or minimizing the action of plant hormone,
ethylene has been an attractive target area of study for the postharvest sci-
entist. This is due to the fact that these technologies can reduce wastage
and postharvest loss of horticultural produce. This chapter outlines the
progress that has been made in our understanding of ethylene signaling in
fruits and vegetables in the recent past.

11.2  Ethylene Physiology in Climacteric

and Nonclimacteric Fruits
Fruits are classically divided into two groups depending on their different
ripening mechanisms (Lin et al., 2009). These include climacteric (toma-
toes, apples, avocados, bananas, etc.), in which ripening is accompanied
by a peak in respiration and a concomitant burst of ethylene, and noncli-
macteric (strawberries, citrus, pineapples, etc.), in which respiration shows
no dramatic change and ethylene production remains at basal levels. The
significance of respiratory climacteric is still poorly understood since non-
climacteric fruits undergo ripening even in the absence of the respiratory
climacteric phenomena. In addition, both climacteric and nonclimacteric
fruits undergo similar physicochemical changes during ripening in terms

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of color, texture, softening, sugar metabolism, synthesis of aroma volatiles,

and increased susceptibility to pathogens (Cara and Giovannoni, 2008;
Seymour et al., 2013b). Similarly, the genes regulating ripening in both
types of fruits tend to exhibit common altered expression patterns, thus
supporting the hypothesis that common ethylene-dependent and ethylene-
independent regulation pathways in both fruits coordinate fruit maturation
processes.
A great deal is known regarding ethylene biosynthesis, ethylene per-
ception and signal transduction, and downstream gene regulation in the
ripening of climacteric fruits (Lin et al., 2009). However, understanding
of the regulatory mechanism of ripening in nonclimacteric fruits and the
upstream regulation of ethylene in climacteric fruits remains elusive.
In fruit and vegetable species, tomato has emerged as the most useful
model to date, due to its commercial importance; ease of genetic manipu-
lation; rapid life cycle; year-round nonseasonal greenhouse fruit produc-
tion; rich source of genetic and biochemical information; relatively small
genome; well-characterized single gene ripening mutants, such as  never-
ripe (nr), non-ripening (nor), ripening-inhibitor (rin), and green-ripe (gr); and
the availability of detailed genetic maps, expressed sequence tag (EST)
collections, microarray chips, and full-length cDNA collections (Alexander
and Grierson, 2002; Giovannoni, 2007; Klee and Giovannoni, 2011; Seymour
et al., 2013a). Consequently, much of our understanding of ethylene signal-
ing in fruit and vegetable species comes from studies on tomato.

11.3  Ethylene Signaling Components in Fruits

After its biosynthesis, ethylene is perceived by a family of copper binding
membrane-associated receptors. The receptors predominantly reside in a
site that is not typical for receptor ligand binding, the endoplasmic retic-
ulum (ER) membrane. However, given that the ethylene is gaseous and
can freely diffuse in both aqueous and lipid environments of the cell, this
localization of the receptors might facilitate interactions with other cellu-
lar components and enable signal integration with other pathways (Ju and
Chang, 2012). A family of seven genes encoding tomato ethylene receptors
have been isolated and characterized in tomato fruit, including LeETR1,
LeETR2, NR, LeETR4, LeETR5, LeETR6, and LeETR7 (O’Malley et al.,
2005; Kevany et al., 2007; Klee and Giovannoni, 2011; Seymour et al., 2013a;
Gapper et al., 2013). The LeETR1, LeETR2, and NR (never-ripe; also called
LeETR3) are classified into the subfamily I group of receptors, whereas
LeETR4, LeETR5, and LeETR6 are subfamily II members. Subfamily I eth-
ylene receptors have three trans-membrane domains and a well-conserved
histidine kinase domain. On the other hand, subfamily II receptors contain
a putative signal peptide in addition to the three conserved trans-membrane

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domains and a histidine kinase domain that lacks one or more elements that
are necessary for catalytic activity. The recently released tomato genome
sequence indicates that the seven ethylene receptors comprise the entire
inventory of tomato ethylene receptors.
The proteins encoded by these genes are structurally diverse and, at
worst, are less than 50% identical. The mRNA expression patterns of the
tomato ethylene receptors have been detected in all tissues analyzed, but
they present distinct expression patterns throughout development and in
response to differing environmental stimuli. The transcript levels of LeETR1
and LeETR2 are constitutive in all tomato tissues throughout development,
while NR, LeETR4, LeETR5, and LeETR6 exhibit increased levels in repro-
ductive tissues (flowers and fruit), with a significant increase of NR, LeETR4,
and LeETR6 in ripening fruits (Kevany et al., 2007). It is plausible that the
increased ethylene evolved at the onset of ripening in tomato is responsible
for the increased transcription of NR, LeETR4, and LeETR6 receptors, since
these three receptor genes are by far the most highly expressed in ripening
fruits. Together, these results indicate that subfamily II tomato receptors
may have far more important functions. The LeETR4, which has been dem-
onstrated to be a critical receptor for tomato fruit ripening, is multiply phos-
phorylated in vivo, and the phosphorylation level is dependent on ripening
stage and ethylene action (Kamiyoshihara et al., 2012).
The ethylene receptors transmit the signal to the downstream Raf
kinase–like protein, the constitutive triple response1 (CTR1) protein. The
subfamily I tomato receptors LeETR1, LeETR2, and NEVER-RIPE (NR)
can interact with multiple LeCTRs (1, 3, and 4) (Zhong et al., 2008). As
with the ethylene receptors, all tissues evaluated expressed CTR1 genes,
and their transcript differentially increased depending on tissue (Adams-
Phillips et al., 2004). The LeCTR1 is the highest expressed of the family
members in fruit, and its abundance is induced by exogenous ethylene appli-
cation and ripening. Genetic studies have illustrated that in the absence of
ethylene perception, the receptors repress ethylene responses by activat-
ing CTR1; binding of ethylene inactivates ethylene receptor signaling, and
CTR1 is consequently inactive, thereby leading to ethylene responses. The
molecular mechanism for how the receptors control CTR1 activity is yet to
be elucidated (Zhong and Chang, 2012).
The next component downstream of CTR1 is Ethylene Insensitive 2
(EIN2), a central positive regulator of ethylene responses residing in the
ER membrane (Bisson et al., 2009; Bisson and Groth, 2011). EIN2 encodes
a protein with similarity to the Nramp family of metal ion carriers and
for which loss-of-function mutations display ethylene insensitivity. EIN2
expression in tomato shows an increase at the onset of ripening (J. Wang
et al., 2007). The expression of LeEIN2 remains the same at different stages
of fruit development and is independent of ethylene. In addition to transcrip-
tional control of EIN2 mRNA accumulation, it is likely EIN2 is also a target

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for protein turnover via the 26S proteosome. EIN2 is thought to play an
important role in the signal transduction of other hormones in addition to
ethylene, such as abscisic acid, auxin, cytokinin, and jasmonate, and thus
may represent a point of cross talk between multiple hormone signaling
pathways (Zhu et al., 2006; Cara and Giovannoni, 2008).
Downstream of EIN2, localized in the nucleus, is a family of short-lived
proteins that act as positive regulators of the ethylene signaling pathway
termed EIN3 and EIL (or EIN3-like) transcriptional regulators. Four EIL
genes have been cloned from tomato, and EIN3/EILs function as dimers,
and at least in the case of the tomato, EIL1, a mutation at a conserved phos-
phorylation site, disrupts fluorescence signal in a tobacco BiFC system, as
well as abolishes the activity of the respective trans-gene in tomato plants
(Li et al., 2012). EIN3/EILs are positive regulators of the ethylene signaling
pathway that act as trans-activating factors to trigger ethylene responses
mainly via the regulation of ethylene response factor (ERF) genes known
to be their downstream targets (Li et al., 2012).
The ethylene signaling cascade ends with a family of transcription
factors, the ERFs. ERF-type transcription factors are specific to plants and
belong to the APETALA2 (AP2)/ERF family. Proteins encoded by this
family have a highly conserved DNA binding domain known as the AP2
domain. The SlERF6 has been defined as a negative regulator of ethylene
and carotenoid biosynthesis in maturing tomato fruit (Lee et al., 2012).
Five tomato ERF genes (LeERF1–4 and LeERF3b) have been described to
date, and the transcript accumulation studies indicated a specific pattern of
expression for each gene, with LeERF2 (Sl-ERF2) and LeERF3b display-
ing ripening-associated transcript accumulation (Yang et al., 2010). While
LeERF2 expression increased during fruit ripening, LeERF3b accumu-
lated before and declined sharply after the onset of ripening. The LeERF2
transcripts were not detected in the tomato ripening mutants Nr, rin, and
nor. In contrast, LeERF3b mRNA is increased in low-ethylene tomato fruit
containing an ACC oxidase sense-suppression trans-gene and in Nr mutant
fruit (Chen et al., 2008).

11.4  Analyses of Ethylene Signal

Components in Other Fruit Species

11.4.1  Climacteric Fruits

Characterization of ethylene receptor genes provides clues to understand
how plants regulate their ethylene sensitivity. The ethylene receptors have
been isolated and characterized in a number of climacteric and noncli-
macteric fruits and exhibit different expression patterns during ripening.

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The melon fruit is second to tomato when it comes to research work car-
ried out on ethylene perception. Melon fruit is ideal for these studies due
to its development having three distinct stages, phases I–III. The flesh,
embryo, placenta, and seeds are well ordered; the fruit development can
be clearly divided into ethylene-insensitive and ethylene-sensitive stages;
and the developing fruit has a lower sensitivity to ethylene than does
the ripening fruit (reviewed in Ezura and Owino, 2007). In muskmelon,
Cm-ERS1 mRNA increased slightly in the pericarp of fruit during ripen-
ing, followed by the marked increase of Cm-ETR1 mRNA, which paral-
leled climacteric ethylene production. The increase of Cm-ERS1 mRNA
at a low concentration of ethylene before the increase of Cm-ETR1 mRNA
and ethylene production indicates that Cm-ERS1 may be sensitive to a
low concentration of ethylene, whereas Cm-ETR1 may be involved in
the response at a high concentration of ethylene. The melon receptor
Cm-ERS1 was determined to localize to the ER, and its topology is such
that it spans the membrane three times, with its N-terminus facing the
luminal space and the large C-terminal portion lying on the cytosolic side
of the ER membrane (Ma et al., 2006).
In mango MiETR1 mRNA level increased in both mesocarp and seed
tissues. However, in those same tissues, MiERS1 transcript accumulation
was barely detected (Ish-Shalom et al., 2011; Contreras-Vergara et al., 2012).
Three ethylene receptor genes, PeETR1, PeERS1, and PeERS2, have been
isolated from passion fruit. The PeETR1 and PeERS1 transcripts are more
elevated in arils than in seeds, while PeERS2 was detected only in the arils
of ripe purple fruit, suggesting that PeERS2 might play a role in repressing
ethylene responses at later developmental stages after fruit ripening has
been completed (Mita et al., 2002).
The avocado PA-ERS1 mRNA increased gradually from the day of har-
vest and did not change significantly until the climacteric peak, when it was
hyperinduced. 1-MCP, however, suppressed the accumulation of PA-ERS1
to basal levels, suggesting that the stimulated induction of PA-ERS at the
climacteric peak may be a mechanism by the avocado fruit to dissipate the
high levels of autocatalytic ethylene (Owino et al., 2002). The avocado seed
has been shown to be involved in the regulation of ethylene responsive-
ness during ripening and contributes to the delay of the climacteric peak in
mature seeded fruits (Hershkovitz et al., 2010). The seedless fruit ripened
more quickly, both at ambient temperature and in cold storage, than seeded
ones. The faster ripening was evident by an early onset of climacteric eth-
ylene production, and the expression of PaETR, PaERS1, and PaCTR1
increased in parallel with the onset of the ethylene burst in seedless fruit,
whereas PaETR increased predominantly in seeded ones. There were also
clear differences between seeded and seedless fruit in the mRNA accu-
mulation patterns of the PaCTR1 gene in response to endogenous or exog-
enous ethylene; PaCTR1 was dramatically induced by exogenous ethylene

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only in seeded fruit, but endogenous ethylene induced it in seedless fruit

(Hershkovitz et al., 2011). Chilling of avocado fruit (Persea americana cv.
‘Arad’) in the orchard induced fruit ripening and a parallel increase in eth-
ylene biosynthesis and receptor genes’ expression during shelf life. The
transcripts of both PaETR and PaERS1 in chilling stressed fruit on the
tree were found to parallel ethylene production and were 25 times higher
in stressed fruit than the control. However, PaCTR1 transcript levels were
less affected by chilling stress (Hershkovitz et al., 2009).
Ethylene perception has also been described to be involved in apple
fruitlet abscission and early development (Dal Cin et al., 2005). The apple
MdETR1 and MdERS1 gene expression patterns were tissue specific, with
MdETR1 transcripts being more abundant in the peduncle, abscission
zone, and seed than in the cortex of early developing fruits (nonabscising
fruitlets), even though the expression always remained at a steady-state
level. The ethylene receptors ETR2 and ETR5, along with the ethylene
control element, were isolated from the summer apple ‘Sunrise’ (SR)
and late season ‘Golden Delicious’ (GD). The patterns of four ethylene
receptors (ETR1, ETR2, ETR5, and ERS1) and CTR1 and EIN2 were
more or less similar in the two apple cultivars (Wiersma et al., 2007).
MdERS1 and MdERS2 transcription increased rapidly after harvest
in control ‘Orin’ and ‘Fuji’ apples, but in 1-MCP-treated fruit, increases
were delayed for 30  days. However, MdERS1 and MdERS2 protein lev-
els behaved differently. MdERS1 decreased gradually in both the control
and 1-MCP treatments. MdERS2, however, increased gradually in control
‘Fuji’ and remained steady in 1-MCP-treated ‘Fuji’, but remained low in
‘Orin’ (Tatsuki et al., 2009). Ethylene response factors and their expres-
sion patterns have been determined in ripening apple (Malus × domestica
Borkh. cv. ‘Golden Delicious’) fruit (A. Wang et al., 2007). MdERF1 was
expressed mainly in ripening fruit, although a small degree of expression
was also observed in nonfruit tissues, whereas MdERF2 was expressed
exclusively in ripening fruit and its transcription was regulated positively
by the ethylene signaling system.
In peach, Pp-ETR1 appeared to be constitutive and ethylene inde-
pendent during fruit development and ripening, while Pp-ERS1 transcripts
increased during fruit ripening, and its expression appeared to be upreg-
ulated by propylene treatment (Rasori et al., 2002). Application of ethyl-
ene inhibitor 1-MCP delayed fruit ripening and ethylene evolution and
concurrently downregulated Pp-ERS1, while Pp-ETR1 transcription was
unaffected. 1-MCP action was rapidly abolished after moving fruits to air,
when a rapid stimulation of ethylene evolution and a concurrent increase of
Pp-ERS1 mRNAs were observed.
Cold treatment leads to a gradual increase in ethylene production,
but there is a commensurate increase in receptor expression in ‘Passe-
Crassane’ pear fruit. The Pc-ETR1a mRNA accumulation was upregulated

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by cold and during ripening. Pc-ERS1a and Pc-ETR5 were less affected by
cold treatment, but all increased during post–cold treatment, ethylene-
dependent ripening (El-Sharkawy et al., 2003). A sharp peak of Pc-ETR1a
and Pc-ERS1a mRNA accumulation was observed during ripening in the
early season pears, in contrast to the gradual increase seen in ‘Passe-
Crassane’ (PC) fruit. A more pronounced difference between early cultivars
and PC fruit can be seen in the behavior of Pc-ETR5, a transcript accumula-
tion. Transcript levels for Pc-ETR5 diminish sharply before and during the
ethylene climacteric and ripening of early season pear fruit, whereas in PC
fruit they increase sharply. This suggests that a decrease in the expression
of a negative regulator could result in an increase in ethylene sensitivity
early in the ripening phase of early fruit development. However, given the
potential for redundancy in the ethylene receptor family, it remains to be
determined whether reduced levels of Pc-ETR5 affect the overall ethylene
sensitivity of early pear fruit (El-Sharkawy et al., 2003).
Three ethylene receptors, DkERS1, DkETR1, and DkETR2, have been
isolated and their expression determined during ripening of persimmon
fruit (Pang et al., 2007). The DkETR1 mRNA is constitutively expressed
during all stages of fruit ripening and is ethylene independent. Conversely,
DkERS1and DkETR1 mRNAs correlated with ethylene production during
fruit development and ripening and were induced by ethylene. The DkERS1
protein decreased gradually prior to fruit maturation and reached its lowest
level at the ripening stage, when ripening-related ethylene was produced,
suggesting the involvement of DkERS1 in ethylene perception during fruit
ripening.
Ethylene signaling elements have been isolated and characterized
in two Japanese plum cultivars, ‘Early Golden’ (EG), with normal cli-
macteric pattern, and ‘Shire’ (SH), with suppressed climacteric pattern
(El-Sharkawy et al., 2007, 2009). These included two receptors (ETR1-
like proteins Ps-ETR1 and Ps-ERS1), one CTR1-like protein (Ps-CTR1),
and an ethylene-responsive element binding factor (Ps-ERF). During the
early stages of development, all four ethylene components were downregu-
lated in an ethylene-independent manner. However, Ps-ETR1 and Ps-CTR1
transcripts accumulated during later stages in an ethylene-independent
manner. Ps-ETR1 was constitutively expressed during development, with
further increased expression during fruit ripening. Ps-ERS1 and Ps-ERF1
transcript levels increased sharply with the climacteric peak of EG fruit in
an ethylene-dependent manner. However, in the SH cultivar, the expression
was constitutive and low.
Five ethylene receptors (AdERS1a, AdERS1b, AdETR1, AdETR2, and
AdETR3), two CTR1-like genes (AdCTR1 and AdCTR2), and an EIN3-like
gene (AdEIL1) have been isolated from kiwifruit. The gene was found to be
fruit specific, and all were differentially expressed among various kiwifruit
tissues (Yin et al., 2008).

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11.4.2  Nonclimacteric Fruits

In contrast to the great deal of information regarding the ethylene recep-
tors in climacteric fruits, much less is known about nonclimacteric fruits.
At present, no single growth regulator appears to play a positive role analo-
gous to the role played by ethylene in the ripening of climacteric fruits.
Nonclimacteric fruits are also able to synthesize ethylene, and in some
cases it has been seen that ethylene can hasten the postharvest deteriora-
tion; the possible involvement of this hormone in the ripening of the noncli-
macteric fruits has been studied in different laboratories. However, in spite
of the many efforts, no results have been obtained that can demonstrate a
clear relation between ethylene and the ripening of these fruits.
During development of male and female flowers of Curcubita pepo, the
expression of both the CTR1 homologs increased along with ethylene pro-
duction in flowers, indicating that both genes are upregulated by ethylene
in flowers, but not in seedlings and leaves (Manzano et al., 2010).
Three ethylene receptors were isolated and expression patterns
determined in strawberry fruits (Trainotti et al., 2005). The FaEtr1 mRNA
was low in flowers, but showed an increase in the small green fruits, and a
subsequent decrease in the large green fruits that was followed by a steep
increment that continued throughout the ripening phase. The FaErs1
mRNA was very high in flowers, but steadily decreased to reach a mini-
mum in the large green fruits. Afterwards, it increased again until the rip-
ening was completed. On the other hand, FaEtr2 mRNA increased about
threefold to reach a maximum in the white fruits. Afterwards, although a
slight decrease was observed, the transcript amount remained high in the
red fruits, at well over twice that of the small green fruits. The FaEtr1 and
FaEtr2 genes were more responsive to ethylene in the white fruits, while
FaErs1 was highly responsive to ethylene at the red stage. This study sug-
gested that ethylene receptors might have a physiological role in the ripen-
ing of nonclimacteric strawberries. The EIL-like genes, ethylene-mediated
signaling components, were induced in pepper fruit (Lee et al., 2010), sug-
gesting common systems for downstream signal transduction in climac-
teric and nonclimacteric fruits, which in the latter occurs in the absence of
ethylene biosynthesis.
Four ethylene receptors have been isolated from grapes, namely
VvETR1, VvETR1, VvERS1, and VvEIN4. Other components of the ethyl-
ene signaling transduction identified in grapes include VvRTE1, VvCTR1,
and VvEIN3 (Chervin and Deluc, 2010). Exogenous ethylene application
resulted in elevated expression of VvETR2 and VvCTR1 mRNAs. Ethylene
antagonist 1-MCP application, on the other hand, resulted in increased tran-
script levels of VvEIN3 after 9 weeks and VvRTE1 after 7 weeks. However,
no significant change was observed in VvEIN4 transcript accumulation.

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Taken together, these observations prompt the prevalence of a common

regulatory pathway in climacteric and nonclimacteric fruit (Chervin and
Deluc, 2010).
Even though citrus are nonclimacteric fruits, exogenous ethylene is
able to stimulate ripening by accelerating respiration and inducing pigment
changes of peel chlorophyll degradation, as well as carotenoid biosynthesis.
In young ‘Valencia’ fruitlets, CsERS1 expression was detected in fruits on
tree immediately after harvest and was further induced in the subsequent
days (Katz et al., 2004). The CsERS1 expression was slightly induced by
ethylene treatment and reduced by 1-MCP treatment in young fruitlets. The
CsETR1 expression was constitutive in young fruitlets, was not affected by
detachment from the tree, and was ethylene independent. In mature fruit,
the expression of both CsERS1 and CsETR1 genes was constant and was
not affected by either 1-MCP or propylene treatments. The differences in
the expression of CsERS1 between young fruitlets and mature fruit suggest
that CsERS1 may modulate the differential sensitivity to ethylene in fruit-
lets versus mature citrus fruit.
The expression levels of ethylene subfamily II receptors CaETR4 and
CaETR5 in nonclimacteric capsicum were significantly lower in 1-MCP-
treated samples than in the control (Aizat et al., 2013). Subfamily II CaETR4
may also be considered to be the major ETR isoform expressed in capsicum
due to its higher overall relative expression during ripening. The expres-
sions of all four ETR isoforms after ethylene exposure were also constantly
compared to the control.

11.5  Transcriptional Regulators of Fruit Ripening

The detailed molecular analyses of ethylene synthesis and signal trans-
duction during ripening suggest the presence of additional regulatory sys-
tems that coordinate the ethylene burst and production of this necessary
ripening hormone. Several tomato ripening mutants have been collected
from production fields or in breeding programs. These include pleiotropic
ripening mutations, such as colorless non-ripening (Cnr), ripening-inhibitor
(rin), never-ripe (Nr), green-ripe (Gr), and high-pigment (hp-1 and hp-2), all
of which have now been isolated either by positional cloning or by genetic
mapping of mutant loci and candidate genes (Giovannoni, 2007; Klee and
Giovannoni, 2011). Their subsequent physiological characterization sug-
gests that they may represent defects in ripening regulatory systems,
but share common physiological characteristics. These tomato mutants
fail to display normal ripening phenotypes, including ethylene synthesis,
increased respiration, carotenoid accumulation, softening, and aroma vola-
tile production. These fruits, from the developmental process to the mature
green stage, (1) attain full size but do not ripen, (2) lack the climacteric

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rise in respiration and do not produce ripening-associated ethylene, (3) do

not ripen even when subjected to exogenous ethylene, but (4) respond to
ethylene in other tissues and fruit in which ethylene is responsive. These
characteristics suggest that all three mutations impact the central ripening
phenomena necessary for the induction of ethylene, in addition to activities
for which ethylene alone cannot compensate.
There is only one described rin mutation, and the rin locus encodes
a MADS-box transcription factor termed RIN-MADS, which is induced
at the onset of ripening (Qin et al., 2012). MADS-box genes are devel-
opmental regulators conserved in eukaryotes and associated with floral
development in plants, including members of the ABC model of floral
development. The ABC model, explains the interaction of three classes of
homeotic genes acts in concert to direct the development of floral organs.
In this classical model, Arabidopsis thaliana A-class genes APETALA1
(AP1) and AP2 confer sepal identity in the first floral whorl. Their activity
overlaps with B-class genes APETALA3 (AP3) and PISTILLATA (PI ) in
the second whorl, which develops into petals. AP3, PI and the C-class gene
AGAMOUS (AG) specify stamen identity in whorl three, while AG alone
in whorl four promotes carpel development. Subsequent cloning of the
ABC genes showed that AP1, AP3, PI and AG all encode MADS domain
proteins, as do the SEPALLATA (SEP) genes, which encode obligatory
co-factors for the homeotic proteins (Wollmann et al., 2010). The tomato
LeMADS-RIN is a member of the SEP4 group of the SEPALLATA (SEP)
class of transcription factors, which are expressed during ripening of a
wide range of fleshy fruit–bearing species (Klee and Giovannoni, 2011;
Vrebalov et al., 2002).
A SEP1/2-like gene in strawberry, a nonclimacteric fruit, is involved
in the development and ripening of the achenes and swollen receptacle, and
silencing of this gene resulted in altered receptacle development, a failure of
the achenes to degreen, and slow ripening in the receptacle (Seymour et al.,
2011). The identification of a strawberry homolog of RIN-MADS indicates
that transcriptional control of ripening is conserved among climacteric and
nonclimacteric species. In banana (Musa AAA group), a SEP3 homolog is
likely to act upstream of the increase in ethylene production in a way similar
to that of RIN (Elitzur et al., 2010). These results point to possible conserva-
tion across taxa of the families of transcription factors involved in the high-
level regulatory network controlling ripening. The tomato AGAMOUS (AG)
ortholog represents the AGAMOUS C-type lineage in the model based on
the formation MADS protein complexes. MADS-box paralogs identified in
tomato (TAG1, TAGL2, TAGL11, TAGL12, TAGL1, TDR6, and TDR4) have
been suggested to be involved in fruit development (Giovannoni, 2007). The
tomato HB-1 encodes a putative HD-Zip homeobox protein that binds to the
promoter of ACC oxidase (LeACO1), a key enzyme in the ethylene biosyn-
thetic pathway (Lin et al., 2008). The rin and Cnr genes encode MADS-box

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and SPBP transcription factors, respectively, and are necessary for ripen-
ing control (Manning et al., 2006).
Repression of TAGL1, MADS-RIN, CNR-SBP, or HB-1 is similar in
that all result in low-ethylene, nonripening fruit, suggesting that all four
genes operate upstream of ethylene biosynthesis and have some functions
that are ethylene independent. HB1 regulates climacteric ethylene through
direct regulation of ACO1, while TAGL1 regulates ethylene (either directly
or indirectly) through regulation of ACS2 with no effect on ACO1. CNR-SBP
mRNA accumulation is reduced in the rin mutant, suggesting that MADS-
RIN has a positive regulatory role on CNR-SBP expression (Vrebalov et al.,
2009). It is also noteworthy that (rin/Rin) hybrids form the basis for most
present-day production of slow-ripening, long-shelf-life, fresh-market toma-
toes demonstrating the commercial importance of the rin mutation.
Arlequin (Alq) is a semidominant T-DNA tomato mutant that exhibits
developmental changes affecting flower and fruit ripening. Genetic, molec-
ular, and functional analyses of the Alq T-DNA mutant indicated that it cor-
responds to the TAGL1 gene as a key MADS-box regulator of reproductive
development in tomato (Seymour et al., 2008; Giménez et al., 2010; Pineda
et al., 2010). Analysis of ALQ/TAGL1 demonstrated that it acts upstream
of ethylene-related genes, though independently to the ripening pathway
regulated by RIN, and regulates flower and fruit development, and there-
fore cannot be considered a specific fruit ripening gene. The ALQ/TAGL1
might act as a linking factor between flower development and fruit ripen-
ing networks, allowing the reproductive development to be successfully
completed.
Additional MADS-box genes have also been reported for other fruits;
however, most of them were suggested to be involved in early fruit devel-
opment. In peach, two MADS-box genes similar to TAG and TAGL1 have
been cloned, and their expression in tomato shows that they might be
responsible for fruit development (Tadiello et al., 2009). MADS-box genes
have also been isolated from Chinese pear, and there was no difference
in their expression in climacteric and nonclimacteric cultivars (Yamane
et al., 2007). Whether or not these genes are specifically involved in rip-
ening remains to be determined in these fruits. In ‘Grand Nain’ bananas,
MaMADS2 is the most likely candidate that serves as an upstream regu-
lator of ripening, since it is not affected by ethylene (Elitzur et al., 2010).
MuMADS1 and a ubiquitin-activating enzyme E1 gene fragment (MuUBA)
were co-regulated by ethylene, suggesting that it might play an important
role in postharvest banana fruit ripening (Liu et al., 2013).
An AGAMOUSMADS-box protein predominantly accumulated in
the climacteric banana fruit pulp and also in female flower ovary suggests
involvement in banana fruit ripening and floral reproductive organ devel-
opment (Roy Choudhury et al., 2012). A ripening-related homolog of the
tomato RIN gene has been isolated in strawberry, which increases neither

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

respiration nor ethylene production during ripening and is largely unre-

sponsive to inhibitors of ethylene action, such as 1-methylcyclopropene
(1-MCP). This is consistent with the hypothesis that such regulators might
be conserved between both climacteric and nonclimacteric fruits. Thus,
RIN and CNR may be good candidates for controlling ripening and pro-
longing shelf life in most produce more than is currently possible via eth-
ylene control alone. The studies outlined above demonstrate that TAGL1,
MADS-RIN, CNR-SBP, and HB-1 are all positive transcriptional regulators
of ripening.
The APETALA2 genes belong to the AP2/ERF (Ethylene Response
Element) family that are mostly associated with responses to biotic and
environmental stress and encode putative transcription factors (Kim et al.,
2006). Characterization of a tomato APETALA2 gene (SIAP2a) demon-
strated its negative role in influencing the ripening process by modulating
the hormone ethylene and carotenoid pigmentation essential for tomato
fruit color and nutrient quality (Chung et al., 2010; Karlova et al., 2011). The
identification of SIAP2a as a negative transcriptional regulator of fruit rip-
ening presents an additional biotechnological tool for modifying the quality
and nutritional value of fruit crops.

11.6  Conclusions and Future Perspectives

Based on dedicated studies on ethylene signaling, it can be deduced
that different means of interplay between ethylene and other signals
make the ethylene signaling pathway an open system that is amenable
to regulation executed by various developmental and environmental
cues. The new advances in fruits and vegetables can be summarized
in a linear model. The recently released tomato genome sequence indi-
cates that there are seven ethylene receptors that comprise the entire
inventory of tomato ethylene receptors. Among these receptors, sub-
family 2 tomato receptors may have far more important functions than
subfamily 1; however, subfamily 1 tomato receptors can interact with
multiple LeCTRs (1, 3, and 4), and the molecular mechanism for how the
receptors control CTR1 activity is yet to be elucidated. The next compo-
nent in the pathway, the EIN2, is thought to play an important role in the
signal transduction of other hormones in addition to ethylene, such as
abscisic acid, auxin, cytokinin, and jasmonate, and thus may represent
a point of cross talk between multiple hormone signaling pathways. The
EIN3/EILs are positive regulators of the ethylene signaling pathway
that act as trans-activating factors to trigger ethylene responses mainly
via the regulation of ethylene response factor (ERF) genes known to
be their downstream targets. The ethylene signaling signal terminates

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 ET H Y LENE SIGNAL TRANS D U C TION

with a family of transcription factors, the ERFs; however, even though

it is clear that the majority of well-characterized responses to this hor-
mone are mediated by the canonical ethylene signaling pathway in a
number of the fruit and vegetable species studies described above,
there is the possibility of the existence of alternative cascades that skip
one or several of the classical ethylene signaling components. These
other parallel cascades are the focus of future research in ethylene
signaling. Tomato has been the source of the natural epigenetic muta-
tions characterized to date. The characteristics of at least three muta-
tions indicate that they impact central ripening phenomena necessary
for induction of ethylene, in addition to activities for which ethylene
alone cannot compensate. It is yet to be elucidated, however, how these
epigenetic variations are responsible for controlling important quality
traits in other fruit and vegetable species.

References
Adams-Phillips, L., Barry, C., Kannan, P., Leclercq, J., Bouzayen, M., and
Giovannoni, J. 2004. Evidence that CTR1-mediated ethylene signal
transduction in tomato is encoded by a multigene family whose mem-
bers display distinct regulatory features. Plant Molecular Biology 54,
387–404.
Aizat, W.M., Able, J.A., Stangoulis, J.C., and Able, A.J. 2013. Characterisation
of ethylene pathway components in non-climacteric capsicum. BMC
Plant Biology 13, 191.
Alexander, L., and Grierson, D. 2002. Ethylene biosynthesis and action in
tomato: A model for climacteric fruit ripening. Journal of Experimental
Botany 53, 2039–2055.
Bisson, M.M., and Groth, G. 2011. New paradigm in ethylene signaling. Plant
Signaling and Behavior 6, 164–166.
Bisson, M.M.A., Bleckmann, A., Allekotte, S., and Groth, G. 2009. EIN2, the cen-
tral regulator of ethylene signaling, is localized at the ER membrane where
it interacts with the ethylene receptor ETR1. Biochemical Journal 424, 1–6.
Bleecker, A.B., and Kende, H. 2000. Ethylene: A gaseous signal molecule in
plants. Annual Review of Cell and Developmental Biology 16, 1–18.
Cara, B., and Giovannoni, J.J. 2008. Molecular biology of ethylene during
tomato fruit development and maturation. Plant Science 175, 106–113.
Chang, C., and Bleecker, A.B. 2004. Ethylene biology: More than a gas. Plant
Physiology 136, 2895–2899.
Chen, G., Hu, Z., and Grierson, D. 2008. Differential regulation of tomato eth-
ylene responsive factor LeERF3b, a putative repressor, and the activa-
tor Pti4 in ripening mutants and in response to environmental stresses.
Journal of Plant Physiology 165, 662–670.

353
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Chervin, C., and Deluc, L. 2010. Ethylene signalling receptors and transcrip-
tion factors over the grape berry development: gene expression profil-
ing. Vitis 49, 129–136.
Chung, M.-Y., Vrebalov, J., Alba, R., Lee, J., McQuinn, R., Chung, J.-D.,
Klein, P., and Giovannoni, J. 2010. A tomato (Solanum lycopersicum)
APETALA2/ERF gene, SlAP2a, is a negative regulator of fruit ripening.
Plant Journal 64, 936–947.
Contreras-Vergara, C.A., Stephens-Camacho, N.A., Yepiz-Plascencia, G.,
González-Aguilar, G.A., Arvizu-Flores, A.A., Sanchez-Sanchez, E., and
Islas-Osuna, M.A. 2012. Cloning and expression of ethylene receptor
ERS1 at various developmental and ripening stages of mango fruit.
Genetics and Molecular Research 11, 4081–4092.
Dal Cin, V., Danesin, M., Boschetti, A., Dorigoni, A., and Ramina, A. 2005.
Ethylene biosynthesis and perception in apple fruitlet abscission
(Malusdomestica L. Borck). Journal of Experimental Botany 56, 2995–3005.
Elitzur, T., Vrebalov, J., Giovannoni, J.J., Goldschmidt, E.E., and Friedman, H.
2010. The regulation of MADS-box gene expression during ripening
of banana and their regulatory interaction with ethylene. Journal of
Experimental Botany 61, 1523–1535.
El-Sharkawy, I., Jones, B., Li, Z.G., Lelievre, J.M., Pech, J.C., and Latche, A.
2003. Isolation and characterization of four ethylene perception elements
and their expression during ripening in pears (Pyrus c­ommunis  L.)
with/without cold requirement. Journal of Experimental Botany 54,
1615–1625.
El-Sharkawy, I., Kim, W.S., El-Kereamy, A., Jayasankar, S., Svircev, A.M., and
Brown, D.C.W. 2007. Isolation and characterization of four ethylene
signal transduction elements in plums (Prunus salicina L.). Journal of
Experimental Botany 58, 3631–3643.
El-Sharkawy, I., Sherif, S., Mila, I., Bouzayen, M., and Jayasankar, S. 2009.
Molecular characterization of seven genes encoding ethylene-respon-
sive transcriptional factors during plum fruit development and ripening.
Journal of Experimental Botany 60, 907–922.
Ezura, H., and Owino, W. O. 2008. Melon, an alternative model plant for elu-
cidating fruit ripening. Plant Science 175, 121–129.
Gapper, N.E., McQuinn, R.P., and Giovannoni, J.J. 2013. Molecular and
genetic regulation of fruit ripening. Plant Molecular Biology 82, 575–591.
Giménez, E., Pineda, B., Capel, J., Antón, M.T., Atarés, A., Pérez-Martín,
F., García-Sogo, B., Angosto, T., Moreno, V., and Lozano, R. 2010.
Functional analysis of the Arlequin mutant corroborates the essential
role of the Arlequin/TAGL1 gene during reproductive development of
tomato. PLoS One 5, e14427.
Giovannoni, J.J. 2004. Genetic regulation of fruit development and ripening.
Plant Cell 16 (Suppl.), S170–180.
Giovannoni, J.J. 2007. Fruit ripening mutants yield insights into ripening con-
trol. Current Opinion in Plant Biology 10, 283–289.

354
 ET H Y LENE SIGNAL TRANS D U C TION

Hershkovitz, V., Friedman, H., Goldschmidt, E.E., Feygenberg, O., and

Pesis,  E. 2009. Induction of ethylene in avocado fruit in response to
chilling stress on tree. Journal of Plant Physiology 166, 1855–1862.
Hershkovitz, V., Friedman, H., Goldschmidt, E.E., Feygenberg, O., and
Pesis, E. 2011. Effect of seed on ripening control components dur-
ing avocado fruit development. Journal of Plant Physiology 168,
2177–2183.
Hershkovitz, V., Friedman, H., Goldschmidt, E.E., and Pesis, E. 2010.
Ethylene regulation of avocado ripening differs between seeded and
seedless fruit. Postharvest Biology and Technology 56, 138–146.
Ish-Shalom, M., Dahan, Y., Maayan, I., and Irihimovitch, V. 2011. Cloning
and molecular characterization of an ethylene receptor gene, MiERS1,
expressed during mango fruitlet abscission and fruit ripening. Plant
Physiology and Biochemistry 49, 931–936.
Ju, C., and Chang, C. 2012. Advances in ethylene signaling: Protein complexes
at the endoplasmic reticulum membrane. AoB Plants 2012, pls031, doi:
10.1093/aobpla/pls031.
Kamiyoshihara, Y., Tieman, D.M., Huber, D.J., and Klee, H.J. 2012. Ligand-
induced alterations in the phosphorylation state of ethylene receptors in
tomato fruit. Plant Physiology 160, 488–497.
Karlova, R., Rosin, F.M., Busscher-Lange, J., Parapunova, V., Do, P.T., Fernie,
A.R., Fraser, P.D., Baxter, C., Angenent, G.C., and de Maagd, R.A. 2011.
Transcriptome and metabolite profiling show that APETALA2a is a
major regulator of tomato fruit ripening. Plant Cell 23, 923–941.
Katz, E., Lagunes, P. M., Riov, J., Weiss, D., and Goldschmidt, E. E. 2004.
Molecular and physiological evidence suggests the existence of a sys-
tem II-like pathway of ethylene production in non-climacteric Citrus
fruit. Planta 219, 243–252.
Kendrick, M.D., and Chang, C. 2008. Ethylene signaling: new levels of com-
plexity and regulation. Current Opinion in Plant Biology 11, 479–485.
Kevany, B.M., Tieman, D.M., Taylor, M.G., Cin, V.D., and Klee, H.J. 2007.
Ethylene receptor degradation controls the timing of ripening in tomato
fruit. Plant Journal 51, 458–467.
Kim, S., Soltis, P.S., Wall, K., and Soltis, D.E. 2006. Phylogeny and domain
evolution in the APETALA2-like gene family. Molecular Biology and
Evolution 23, 107–120.
Klee, H.J. 2004. Ethylene signal transduction: Moving beyond Arabidopsis.
Plant Physiology 135, 660–667.
Klee, H.J., and Giovannoni, J.J. 2011. Genetics and control of tomato fruit rip-
ening and quality attributes. Annual Review of Genetics 45, 41–59.
Lee, J.M., Joung, J.-G., McQuinn, R., Chung, M.-Y., Fei, Z., Tieman, D., Klee,
H., and Giovannoni, J. 2012. Combined transcriptome, genetic diver-
sity and metabolite profiling in tomato fruit reveals that the ethylene
response factor SlERF6 plays an important role in ripening and carot-
enoid accumulation. Plant Journal 70, 191–204.

355
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lee, S., Chung, E.-J., Joung, Y.-H., and Choi, D. 2010. Non-climacteric fruit
ripening in pepper: Increased transcription of EIL-like genes normally
regulated by ethylene. Functional and Integrative Genomics 10, 135–146.
Li, J., Li, Z., Tang, L., Yang, Y., Zouine, M., and Bouzayen, M. 2012. A con-
served phosphorylation site regulates the transcriptional function of
ETHYLENE-INSENSITIVE3-like1 in tomato. Journal of Experimental
Botany 63, 427–439.
Lin, Z., Hong, Y., Yin, M., Li, C., Zhang, K., and Grierson, D. 2008. A tomato
HD-Zip homeobox protein, LeHB-1, plays an important role in floral
organogenesis and ripening. Plant Journal 55, 301–310.
Lin, Z., Zhong, S., and Grierson, D. 2009. Recent advances in ethylene
research. Journal of Experimental Botany 60, 3311–3336.
Liu, J.-H., Zhang, J., Jia, C.-H., Zhang, J.-B., Wang, J.-S., Yang, Z.-X., Xu,
B.-Y., and Jin, Z.-Q. 2013. The interaction of banana MADS-box protein
MuMADS1 and ubiquitin-activating enzyme E-MuUBA in postharvest
banana fruit. Plant Cell Reports 32, 129–137.
Ma, B., Cui, M.-L., Sun, H.-J., Takada, K., Mori, H., Kamada, H., and Ezura,
H. 2006. Subcellular localization and membrane topology of the melon
ethylene receptor CmERS1. Plant Physiology 141, 587–597.
Manning, K., Tör, M., Poole, M., Hong, Y., Thompson, A.J., King, G.J.,
Giovannoni, J.J., and Seymour, G.B. 2006. A naturally occurring epigen-
etic mutation in a gene encoding an SBP-box transcription factor inhib-
its tomato fruit ripening. Nature Genetics 38, 948–952.
Manzano, S., Martínez, C., Gómez, P., Garrido, D., and Jamilena, M. 2010.
Cloning and characterisation of two CTR1-like genes in Cucurbita pepo:
regulation of their expression during male and female flower develop-
ment. Sexual Plant Reproduction 23, 301–313.
Merchante, C., Alonso, J.M., and Stepanova, A.N. 2013. Ethylene signaling:
Simple ligand, complex regulation. Current Opinion in Plant Biology 16,
554–560.
Mita, S., Kawamura, S., and Asai, T. 2002. Regulation of the expression of a
putative ethylene receptor, PeERS2, during the development of passion
fruit (Passifloraedulis). Physiologiaplantarum 114, 271–280.
O’Malley, R.C., Rodriguez, F.I., Esch, J.J., Binder, B.M., O’Donnell, P., Klee,
H.J., and Bleecker, A.B. 2005. Ethylene-binding activity, gene expres-
sion levels, and receptor system output for ethylene receptor family
members from Arabidopsis and tomato. Plant Journal 41, 651–659.
Owino, W.O., Nakano, R., Kubo, Y., and Inaba, A. 2002. Differential regulation
of genes encoding ethylene biosynthesis enzymes and ethylene response
sensor ortholog during ripening and in response to wounding in avoca-
dos. Journal of the American Society for Horticultural Science 127, 520–527.
Pang, J.H., Ma, B., Sun, H.-J., Ortiz, G.I., Imanishi, S., Sugaya, S., Gemma,
H., and Ezura, H. 2007. Identification and characterization of ethylene
receptor homologs expressed during fruit development and ripening in
persimmon. Postharvest Biology and Technology 44, 195–203.

356
 ET H Y LENE SIGNAL TRANS D U C TION

Pineda, B., Giménez-Caminero, E., García-Sogo, B., Antón, M.T., Atarés, A.,
Capel, J., Lozano, R., Angosto, T., and Moreno, V. 2010. Genetic and
physiological characterization of the arlequininsertional mutant reveals
a key regulator of reproductive development in tomato. Plant and Cell
Physiology 51, 435–447.
Qin, G., Wang, Y., Cao, B., Wang, W., and Tian, S. 2012. Unraveling the regula-
tory network of the MADS box transcription factor RIN in fruit ripen-
ing. Plant Journal 70, 243–255.
Rasori, A., Ruperti, B., Bonghi, C., Tonutti, P., and Ramina, A. 2002.
Characterization of two putative ethylene receptor genes expressed
during peach fruit development and abscission. Journal of Experimental
Botany 53, 2333–2339.
Roy Choudhury, S., Roy, S., Nag, A., Singh, S.K., and Sengupta, D.N. 2012.
Characterization of an AGAMOUS-like MADS box protein, a probable
constituent of flowering and fruit ripening regulatory system in banana.
PLoS One 7, e44361.
Seymour, G.B., Chapman, N.H., Chew, B.L., and Rose, J.K.C. 2013a.
Regulation of ripening and opportunities for control in tomato and other
fruits. Plant Biotechnology Journal 11, 269–278.
Seymour, G.B., Ostergaard, L., Chapman, N.H., Knapp, S., and Martin, C.
2013b. Fruit development and ripening. Annual Review of Plant Biology
64, 219–241.
Seymour, G., Poole, M., Manning, K., and King, G.J. 2008. Genetics and epi-
genetics of fruit development and ripening. Current Opinion in Plant
Biology 11, 58–63.
Stepanova, A.N., and Alonso, J.M. 2009. Ethylene signaling and response:
Where different regulatory modules meet. Current Opinion in Plant
Biology 12, 548–555.
Tadiello, A., Pavanello, A., Zanin, D., Caporali, E., Colombo, L., Rotino, G.L.,
Trainotti, L., and Casadoro, G. 2009. A PLENA-like gene of peach is
involved in carpel formation and subsequent transformation into a
fleshy fruit. Journal of Experimental Botany 60, 651–661.
Tatsuki, M., Hayama, H., and Nakamura, Y. 2009. Apple ethylene receptor
protein concentrations are affected by ethylene, and differ in cultivars
that have different storage life. Planta 230, 407–417.
Trainotti, L., Pavanello, A., and Casadoro, G. 2005. Different ethylene recep-
tors show an increased expression during the ripening of strawber-
ries: Does such an increment imply a role for ethylene in the ripening
of these non-climacteric fruits? Journal of Experimental Botany 56,
2037–2046.
Vrebalov, J., Pan, I.L., Arroyo, A.J.M., McQuinn, R., Chung, M.Y., Poole,
M., Rose, J., Seymour, G., Grandillo, S., Giovannoni, J., and Irish, V.F.
2009. Fleshy fruit expansion and ripening are regulated by the tomato
SHATTERPROOF gene TAGL1. Plant Cell 21, 3041–3062.

357
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Vrebalov, J., Ruezinsky, D., Padmanabhan, V., White, R., Medrano, D., Drake,
R., Schuch, W., and Giovannoni, J. 2002. A MADS-box gene necessary
for fruit ripening at the tomato ripening-inhibitor (rin) locus. Science
296(5566), 343–346.
Wang, A., Tan, D., Takahashi, A., Li, T.Z., and Harada, T. 2007. MdERFs, two
ethylene-response factors involved in apple fruit ripening. Journal of
Experimental Botany 58, 3743–3748.
Wang, J., Chen, G., Hu, Z., and Chen, X. 2007. Cloning and characterization
of the EIN2-homology gene LeEIN2 from tomato. DNA Sequence 18,
33–38.
Wiersma, P.A., Zhang, H., Lu, C., Quail, A., and Toivonen, P.M.A. 2007. Survey
of the expression of genes for ethylene synthesis and perception during
maturation and ripening of ‘Sunrise’ and ‘Golden Delicious’ apple fruit.
Postharvest Biology and Technology 44, 204–211.
Wollmann, H., Mica, E., Todesco, M., Long, J. A., and Weigel, D. 2010.
On reconciling the interactions between APETALA2, miR172 and
AGAMOUS with the ABC model of flower development. Development
137, 3633–3642.
Yamane, M., Abe, D., Yasui, S., Yokotani, N., Kimata, W., Ushijima, K., Nakano
R., Kubo, Y., and Inaba, A. 2007. Differential expression of ethylene bio-
synthetic genes in climacteric and non-climacteric Chinese pear fruit.
Postharvest Biology and Technology 44(3), 220–227.
Yang, Y., Wu, Y., Pirrello, J., Regad, F., Bouzayen, M., Deng, W., and Li, Z.
2010. Silencing Sl-EBF1 and Sl-EBF2 expression causes constitutive
ethylene response phenotype, accelerated plant senescence, and fruit
ripening in tomato. Journal of Experimental Botany 61, 697–708.
Yin, X., Chen, K., Allan, A.C., Wu, R., Zhang, B., Lallu, N., and Ferguson, I.B.
2008. Ethylene-induced modulation of genes associated with the eth-
ylene signaling pathway in ripening kiwifruit. Journal of Experimental
Botany 59, 2097–2108.
Zhao, Q., and Guo, H.-W. 2011. Paradigms and paradox in the ethylene signal-
ing pathway and interaction network. Molecular Plant 4, 626–634.
Zhong, S., and Chang, C. 2012. Ethylene signalling: the CTR1 protein kinase.
Annual Plant Reviews, The Plant Hormone Ethylene 44, 150.
Zhong, S., Lin, Z., and Grierson, D. 2008. Tomato ethylene receptor–CTR
interactions: Visualization of NEVER-RIPE interactions with multiple
CTRs at the endoplasmic reticulum. Journal of Experimental Botany 59,
965–972.
Zhu, H.-L., Zhu, B.-Z., Shao, Y., Wang, X.-G., Lin, X.-J., Xie, Y.-H., Li, Y.-C.,
Gao, H.-Y., and Luo, Y.-B. 2006. Tomato fruit development and ripening
are altered by the silencing of LeEIN2 gene. Journal of Integrative Plant
Biology 48, 1478–1485.

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Chapter 12

Internal Atmosphere
of Fruits: Role and
Significance in
Ripening and
Storability
Vijay Paul and Rakesh Pandey
Indian Agricultural Research Institute,
New Delhi, India

Abstract 360
12.1 Introduction 361
12.2 Endogenous Volatiles in Fruits 363
12.3 Factors Affecting and Basis of Internal Atmosphere
of Harvested Fruit 363
12.4 Variability in the Internal Atmosphere of Fruits 366
12.5 Influence of the Internal Atmosphere on Ripening
and Related Aspects 367
12.5.1 Ripening 367
12.5.2  Flavor and Aroma 368
12.5.3  Fruit Decay 370

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12.6 Ripening Behavior of Some Fruits under Attached

and Detached Conditions 371
12.6.1 Tomato 371
12.6.2 Melons 372
12.6.3  Sweet Pepper 372
12.6.4 Saskatoon 373
12.7 Role of Some Gases and Endogenous Volatiles
in Fruit Ripening 373
12.7.1 Ethylene 373
12.7.1.1 Role of Ethylene in Ripening
of Climacteric Fruits 373
12.7.1.2 Role of Ethylene in Ripening
of Some Nonclimacteric Fruits 375
12.7.2  Oxygen and Carbon Dioxide 379
12.7.2.1 Low Oxygen 379
12.7.2.2 High Carbon Dioxide 382
12.7.2.3 Ratio of O2 to CO2 383
12.7.3  Ethanol and Acetaldehyde 384
12.7.4  Water Vapors and Water Status in Fruit 385
12.7.5  Salicylic Acid and Methyl Salicylate 387
12.7.6  Jasmonic Acid and Jasmonates 387
12.7.7  Nitric Oxide 388
12.8 Internal Atmosphere of Fruits: Practical Implication
in Ripening and Storability 388
12.9 Conclusions and Future Perspectives 390
References 391

Abstract
Ripening, storability, quality attributes, and postharvest losses in fruits
are interlinked with one another. The postharvest life of a fruit is primarily
determined by various physiological processes and associated metabolic
changes occurring in the fruit. The role of the external atmosphere in reg-
ulating the above processes and changes is relatively better understood.
However, little is known about the overall internal atmosphere of the fruit
and how it influences different aspects of ripening and storability. This
chapter looks into this emerging area: the basic and applied importance
of the internal atmosphere to postharvest physiology and food science and
technology. There are various gases and volatiles that make the internal
­atmosphere of fruits. Their production and diffusion across the fruit tissues

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are governed by many factors. Differences in morphological, a­ natomical,

and microstructural features of fruits are now assuming greater impor-
tance, as they are involved in determining the internal environment of
fruits. As a consequence, there exists variability in the internal atmosphere
of fruits, which is evident not only at the level of different species, but also
within species. Differences in ripening behavior of different fruits under
plant-attached and -detached conditions are also expected in view of the
above. The involvement of some of gases (ethylene, oxygen, and carbon
dioxide) and volatiles (ethanol, acetaldehyde, water vapors and water sta-
tus, salicylic acid and methyl salicylate, jasmonic acid and jasmonates, and
nitric oxide) in the regulation of ripening-related changes, including flavor
and aroma, is described and discussed at the individual as well as at the
interactive level (especially with ethylene). Some examples are presented
wherein endogenous and exogenous volatiles exhibit a positive effect on
the fruit’s storability, quality, and tolerance to biotic and abiotic stresses.
Lastly, a few researchable issues are suggested. The outcome from this area
can supplement the existing storage technologies, and this will be highly
desirable in achieving a more effective and holistic way of the postharvest
management of perishable commodities.

12.1 Introduction
Postharvest physiology, shelf life, and losses in fruits are interlinked and
primarily governed by the last phase of fruit maturation, referred to as
ripening. Fruit ripening involves many physiological, biochemical, and
developmental changes occurring through a coordinated and genetically
regulated program (Stepanova and Alonso, 2005; Barry and Giovannoni,
2007; Bouzayen et al., 2010). Fruits, in general, show two distinctive respi-
ratory patterns during the course of ripening, and on this basis, fruits
are categorized into climacteric and nonclimacteric groups (Biale, 1964;
McMurchie et  al., 1972; Biale and Young, 1981; Abeles et  al., 1992; Paul
et al., 2012). Climacteric fruits show a dramatic increase in the rate of res-
piration during ripening, and this is referred to as climacteric rise. The
rise in respiration is simultaneous, or it is just followed after the rise in
the rate of ethylene production (Burg and Burg, 1962, 1965a; Lelievre
et  al., 1997). The process of ripening can be triggered and also acceler-
ated by exogenous ethylene treatment in climacteric fruits (Tucker, 1993).
The plant hormone ethylene plays a major role in the ripening process
of climacteric fruits, and the presence of ethylene and its perception is
required for the expression of ripening-related genes, even at advance
stages of fruit ripening (Alexander and Grierson, 2002; Hoeberichts et al.,
2002). On the other hand, respiration rates increase at least temporarily

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in nonclimacteric fruits when treated with exogenous ethylene. These

fruits also senescence more rapidly in the presence of ethylene. However,
more recent work c­ arried out in the last 15 years, as reviewed by Paul et al.
(2012), has indicated that ethylene-dependent and ethylene-independent
pathways coexist in both climacteric and nonclimacteric fruits. Findings
showed the fading distinctions between classical patterns of ripening in
climacteric and nonclimacteric fruits. Besides this, low levels of ethylene
are involved in wound healing and responses to various infections in some
fruits of either the climacteric or nonclimacteric group (Van Loon et al.,
2006). In general, the perishability of climacteric fruits is more rapid and
severe than that of the nonclimacteric fruits (Mishra and Gamage, 2007),
basically due to the faster rate of ripening and ripening-related changes in
climacteric fruits.
Postharvest metabolic changes leading to increased respiratory
­activity and transpirational loss of water are the two basic aspects that deter-
mine the storage life and quality of fruits. Approaches such as controlled
atmosphere (CA), modified atmosphere (MA), and modified atmosphere
packaging (MAP) have become the established methods for extending the
postharvest life of fruits (Kader, 2009; Yahia, 2009; Ramayya et al., 2012).
Today, different edible coatings with wide variations in their permeability
to O2, CO2, and water vapors are also available for practical use (Mishra
et al., 2010). It has been observed that the above methods basically modify
the internal gaseous atmosphere of the fruits in favor of a low O2 -to-CO2
ratio (Baldwin et  al., 1999; Elyatem et  al., 1994; Kader, 2009; Nath et  al.,
2011; Yahia, 2009). These methods also regulate ethylene production and
its response (Scully and Horsham, 2008; Kanellis et al., 2009).
At ambient temperature, the internal atmosphere in fruits comprises a
mixture of many gases and volatiles, including O2, CO2, water vapors, ethyl-
ene, alcohols, aldehydes, acetates, esters, ketones, aromatic hydrocarbons,
terpenes, carboxylic acids, sulfur compounds, ammonia, jasmonate, and
salicylates (Baldwin et al., 2000; Pesis, 2005; de Leon-Sanchez et al., 2009).
Several workers have suggested that these gases and volatiles are involved
in regulating ripening, senescence, and related processes (Lougheed et al.,
1987; Toivonen, 1997). In light of this, Saltveit (2003) posed a question: Is it
possible to find an optimal controlled atmosphere for storing fruits and veg-
etables? He emphasized that it is not only the external, but also the internal
environment of the commodity that determines its storability under a given
external environment. This chapter investigates the gases and volatiles
present in the internal atmosphere of fruit and the factors controlling their
production and diffusion through fruit tissue. The role of some of these
gases and volatiles in regulating the ripening and ripening-related changes
in different fruits is described and discussed at the individual as well as the
interactive level.

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12.2  Endogenous Volatiles in Fruits

In plants and plant parts, volatiles are produced in variable amounts at dif-
ferent times and in different tissues (Negre-Zakharov et al., 2009; Defilippi
et al., 2009). Plants use various mechanisms to regulate the production and
levels of these volatiles (Negre-Zakharov et al., 2009; Defilippi et al., 2009).
At present, more than 1000 organic compounds have been reported to be
emitted by plants (Dudareva et al., 2004). The aroma produced by various
fruits during ripening was reviewed (Defilippi et al., 2009; Dudareva et al.,
2004; Pandit et al., 2009). Approximately 400 volatile compounds have been
found in the ripening tomato fruit (Baldwin et al., 1991; de Leon-Sanchez
et al., 2009). The volatiles present in fruits are as follows: ethylene, etha-
nol, acetaldehyde, methanol, acetone, butanol, ethane, hexanol, hexenol,
3-methylbutanal, ethylacetate, propylacetate, butylacetate, propanol,
acetate esters, ethylbutyrate, geraniol, octenal, octenol, citral, terpenes,
carboxylic acids, sulfur compounds, ammonia, jasmonate, benzaldehyde,
and methyl salicylate, besides other types of iso-, sec-, or tert-alcohols,
aromatic hydrocarbons, ketones, esters, aldehydes, and higher carbon
alcohols (Gustafson, 1934; Petro-Turza, 1987; Baldwin et  al., 1991, 2000;
McDonald et  al., 1996; Cadwallader, 2005; de Leon-Sanchez et  al., 2009;
Negre-Zakharov et al., 2009).

12.3  Factors Affecting and Basis of Internal

Atmosphere of Harvested Fruit
The rate of movement of a gas and volatile compound (present within the
plant tissue) depends on the properties of that molecule, the concentration
gradient, and the physical attributes of the intervening barriers (Kader,
1987; Gershenzon et al., 2000; Dudareva et al., 2004). The membranes of the
storage compartment (where the compound exists) or epidermal cell wall
might be differentially permeable to different volatile compounds. Little is
known about metabolite trafficking between various subcellular compart-
ments, the mechanism of the release process, and how these processes
contribute to the regulation of volatile emission (Dudareva et  al., 2004).
Usually, emission of a particular volatile compound into the atmosphere
depends on the rate of its biosynthesis and the rate of its release (Dudareva
and Pichersky, 2000). Formation of volatile compounds is regulated at spa-
tial (Pare and Tumlinson, 1997) and developmental levels (Bouwmeester
et al., 1998; Dudareva and Pichersky, 2000). Further, environmental factors
such as light, temperature, and moisture status can greatly influence the
emission of volatiles (monoterpenes) from the leaves of plant (Staudt and
Bertin, 1998; Gershenzon et al., 2000).

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There are three major routes through which gaseous exchange takes
place for a harvested commodity: the outermost layers (cuticle, cuticular
cracks, and periderm), apertures (stomata and lenticels), and stem scar
region (Solomos, 1987; Ben-Yehoshua and Rodov, 2003). In view of the pres-
ence of cracks on the cuticular layer, the exchange of gases and volatiles
was reported through the cuticular layer, and it is primarily determined
by the extent of cracks present on cuticular layer (Ehret et al., 1993). For
most of horticultural produce, the skin represents the major barrier to
gas exchange (Kader, 1987; Solomos, 1987). The diffusivity of gases for
the fruit flesh is 10–20 times higher than the diffusivity from the skin
(Solomos, 1987; Banks and Nicholson, 2000). In tomato, the stem scar (the
place where the pedicel, along with sepals, connects the fruits to the stem)
is the predominant site for gas exchange (Cameron and Yang, 1982). It was
also demonstrated that 85%–90% of ethylene exchange occurs through this
region of tomato fruit (de Vries et al., 1995). The release of volatiles from
the plant organs with lower rates of transpiration (such as bulky fruits with
well-developed diffusion barriers on the surface, peel, or pericarp) will
be less, and therefore the volatiles can accumulate in the tissues of such
organs. Keeping in view the varietal variation in diffusion barriers and their
characteristics for different plants and plant parts, the accumulation of vola-
tiles would be different, and thereby so would their effect on various physi-
ological processes (Paul et al., 2011; Paul and Pandey, 2014).
Gas diffusion in fruits follows Fick’s law (Burg and Burg, 1965b),
which states that the flux of a gas, diffusing normally through a barrier, is
dependent on the diffusion coefficient and concentration gradient. There
is a morphological and anatomical basis for the gaseous exchange across
the boundaries of harvested fruits (Kader and Morris, 1977; Solomos, 1987;
Kader and Saltveit, 2003a, 2003b). As per Kader and Morris (1977), anatom-
ical features, and not any alterations in biochemical pathways, are the rea-
sons for differences in the diffusion resistance. External (morphological)
and internal (anatomical) features were reported to determine the keep-
ing quality of grapes (Uys, 1974), storability of tomato berries (Niemirow-
Krizsai and Csillag, 1994; Mulas, 1994), and firmness or mechanical
resistance of tomato (Wann, 1996) and olive fruits (Mulas, 1994). In a
study by Bai et  al. (2003), it was shown that ‘Granny Smith’ apples have
relatively few open pores (lenticels) in their peel surface, and therefore they
suffer from low rates of gas exchange; as a result, these fruits are prone
to develop off-flavors after they have been coated with waxes. In contrast,
‘Delicious’ apples have many open lenticels, and they retain sufficient gas
exchange even when coated with waxes. So, waxed ‘Delicious’ apples accu-
mulate only low concentrations of ethanol and off-flavors (Bai et al., 2003).
Likewise, it was reported that Murcott mandarins are much more sensi-
tive to anaerobic stress conditions than Star Ruby grapefruit (Shi et  al.,
2005). Anatomical observations revealed that although the total thickness

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of the peel (comprising the albedo, the white inner layer, and the flavedo
[the c­ olored outer layer]) was greater in grapefruit, it was more condensed
in mandarins. So, it was concluded that mandarins accumulate a larger
amount of acetaldehyde and ethanol after harvest than grapefruit because
of the higher activity of the enzyme alcohol dehydrogenase (ADH) in the
juice vesicles and lower permeability of their peel to gases (Shi et al., 2007).
The extent of diffusibility of gases across the fruit boundaries primar-
ily determines the internal atmosphere of the fruit in terms of the levels of
O2 and CO2 (Ben-Yehoshua et al., 1983; Nuevo et al., 1984). So, the composi-
tion of the internal atmosphere of the fruit is always different from that of the
external atmosphere in which it is kept (Dadzie et al., 1993). The exchange
of gases in fruits through the peel via diffusion from the openings (stomata
and lenticels) is proportional to the difference in the concentrations of gases
across the barrier, total area of the peel, solubility of a gas in peel, solid-
state diffusion coefficient, and total hole area available on the peel surface
(as contributed by openings of stomata and lenticels) (Hagenmaier, 2004,
2005). Gas transport in fruit tissue is governed by diffusion as well as by
permeation. The permeation is basically caused by an overall pressure gra-
dient of a given gas (Ho et al., 2006a). So, a permeation–diffusion–­reaction
model was applied to study gas transport in an intact pear. Permeation was
found to be at a minimum across skin, and it gradually increased for cortex
and vasculature tissues of pear fruit (Ho et al., 2006a).
To a large extent, gas exchange depends on the arrangement pattern
of cells and intercellular spaces (Mendoza et al., 2007). For pear fruit, gas
diffusibility in the vertical axis was higher than that in the equatorial radius
axis (Ho et al., 2006b). In view of this, a very comprehensive model of gas
exchange of pear fruit was proposed for explaining the development of phys-
iological disorder, such as core breakdown and its role in long-term storage
of this fruit (Franck et al., 2007). Later on, Verboven et al. (2008) used high-
resolution phase tomography (making use of synchrotron radiation) to
explore the three-dimensional (3-D) structure and cellular arrangements
of pome fruit tissues in their natural state (i.e., with high water content) up
to a submicrometer resolution. For this study, pome fruits such as apple and
pear were selected because their gas exchange properties have been shown
to be very different and closely related to their storage lives (Schotsmans
et  al., 2004; Ho et  al., 2006a, 2006b; Franck et  al., 2007; Verboven et  al.,
2008; Ho et  al., 2008). Results obtained (Verboven et  al., 2008) revealed
very clearly that the apple fruit had more voids than the pear. The differ-
ences in void fraction (23% for apple cortex and only 5% for pear cortex),
along with the extent of network architectures of voids, explained the bet-
ter ability of tissues to facilitate the gas exchange in apple fruit. This lower
void volume in pear fruit than in apples (Verboven et al., 2008) was able to
explain the earlier findings where pear fruit was found to be more sensi-
tive to physiological disorder, such as internal browning and its relation to

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gas exchange and the availability of internal O2 (Lammertyn et al., 2003;

Franck et  al., 2007; Ho et  al., 2008). Likewise, there is a risk of develop-
ing physiological disorders in the pear fruit during the course of ripening.
This was shown to be due to an increase in respiration, resulting in anoxia
at and near the center of the fruit, even under the recommended storage
conditions (Ho et al., 2010). Very recently, quantification of microporosity
in apple and tomato fruits was done by magnetic resonance imaging (MRI)
for better understanding of the relationship between gas transfer and vari-
ous disorders in fruits during their postharvest life (Musse et al., 2010).

12.4  Variability in the Internal Atmosphere of Fruits

There exists a large variability in the internal atmosphere of fruits belong-
ing to different species, varieties and cultivars, and developmental stages,
sizes, and maturity status (Kader and Morris, 1977; Zagory and Kader, 1988).
Varietal variability in the outer surface morphology, as well as in internal
anatomical features, such as the distribution of trichomes, stomata, and lenti-
cels; the thickness of the cuticle; and the extent of cuticular cracks, could be
the possible reason for the above-mentioned variability in the internal atmo-
sphere of fruits (Kader and Morris, 1977; Zagory and Kader, 1988; Paul and
Srivastava, 2006; Paul et al., 2007, 2010b). Variations in the amount, composi-
tion, and ultrastructure of the cuticular and epicuticular wax among several
apple cultivars were documented (Belding et al., 1998). Apart from this, mor-
phological and mechanical properties of the cuticle, as well as the epidermis,
were subjected to considerable change during growth and ripening of tomato
fruit (Kovacs et al., 1994; Peschel et al., 2003; Bargel and Neinhuis, 2005).
A number of stomata show significant differences among the cultivars of pear
fruit (Kovacs et al., 1994) and sweet cherry (Peschel et al., 2003). Likewise,
varietal variations in the number of lenticels, deposition pattern of the cuticu-
lar and epicuticular wax, amount of cuticle and wax, and internal anatomy
of the peel and exocarp regions were reported in mango fruits (Dietz et al.,
1988; Paul et al., 2007). In tomato fruit, the cuticle appeared to provide an
excellent barrier (Thompson, 2003), and as a result, it may not contribute
significantly to the overall gaseous exchange across the fruit. However,
instead of stomata, trichomes were observed on the surface of tomato fruits,
and trichome base cells are transformed into lenticels during maturation of
the fruit (Blanke, 1986; Paul and Srivastava, 2006). There are varietal differ-
ences in the number of trichomes, tendency of trichomes to get transformed
into lenticels, density of lenticels, and dimension of the stem scar portion of
tomato fruits (Paul and Srivastava, 2006; Paul et al., 2010b). The ratio of O2
to CO2 also gets altered with the growth, development, and maturity of the
fruit (Kader and Morris, 1977; Zagory and Kader, 1988). Gas exchange prop-
erties (specifically due to skin resistance) are not found to be the same with

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the growth and maturity of the fruit. The reason for this may be the changes
that occur in the anatomical properties and features of the fruit itself during
its development and maturation (Longhurst et al., 1994; Kader and Saltveit,
2003a; Bargel and Neinhuis, 2005; Paul et al., 2007; Ho et al., 2008).
Wide differences in the concentrations of volatiles were recorded
in different varieties of apricot (Guichad and Souty, 1988). The levels and
proportions of different volatiles were also found to be responsible for the
characteristic differences in flavor among the varieties of apricot (Guichad
and Souty, 1988). Similarly, Larsen and Poll (1990) found differences in fla-
vor among the 10 raspberry cultivars due to variation in the production of
aroma volatiles. In apple cultivars, wide differences in susceptibility to scald
were associated with the α-farnesene content (Huelin and Coggiola, 1968;
Watkins et al., 1993). Gran and Beaudry (1993) observed wide variability in
the threshold levels of oxygen required (at 0°C) for the induction of anaero-
bic respiration in three apple varieties: 0.7% for ‘Red Delicious’, 1.4% for ‘Red
Fuji’, and about 1.9% for ‘McIntosh’. In this way, variety and storage condi-
tion can influence the degree of accumulation of products of anaerobic res-
piration, such as acetaldehyde and ethanol in tissues. Likewise, out of two
varieties of raspberries (‘Meeker’ and ‘Qualicum’), when harvested at the
red-ripe stage and stored at 1°C with 90% relative humidity (RH) for 7 days,
the variety ‘Qualicum’ was found to be more susceptible to the accumulated
acetaldehyde, ethanol, and ethylacetate in the MAP, with high CO2 (i.e.,
10% CO2) with 5% O2 in comparison with other MAP conditions (6% CO2
with 10% O2 and 3% CO2 with 15% O2) (Toivonen et al., 1999). In tomato fruit,
differences in flavor among different varieties were in part due to the varia-
tion in production levels of aroma volatiles (Brauss et al., 1998). For mango
fruits, volatiles (in different varieties and at different maturity stages) have
been used as a marker for the identification of different maturity stages
in different varieties. So, volatiles can thereby be used in determining the
most optimum maturity stage for harvesting the mango fruits, as this can
result in attaining the best quality of harvested fruits on ripening (Lebrun
et al., 2008; Pandit et al., 2009). In another study, discrimination of 28 apri-
cot cultivars into four distinguishable aroma groups was achieved by ana-
lyzing their volatile constituents (Aubert and Chanforan, 2007).

12.5  Influence of the Internal Atmosphere

on Ripening and Related Aspects

12.5.1 Ripening
During the ripening of tomato fruits, a rise in the endogenous concentrations
of CO2 and ethylene was reported, along with a decrease in the concentration
of O2 (Lyons and Pratt, 1964). The significance of the stem scar region as

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a major site for gaseous exchange of tomato fruit was pointed out by Calbo
et al. (1988) and Yang and Shewfelt (1999). The relationship between ripen-
ing behavior and the stem scar region of tomato fruit in different varieties
was studied by blocking the stem scar region either completely or to different
degrees (Paul et al., 2010b). It was made clear from this study that it is the
degree of blockage of the stem scar region that determines the extent to which
the rate of respiration and ripening are suppressed. In the above studies, the
basic change causing the delay in ripening or suppression of respiration was
primarily due to the lower levels of ethylene and the O2 -to-CO2 ratio within
the fruit. Toivonen (1997) had reviewed the accumulation of nonethylene and
nonrespiratory volatiles (alcohols, aldehydes, jasmonates, terpenes, carbox-
ylic acids, sulfur compounds, and ammonium) and discussed them in terms of
their biological activity in harvested fruits and vegetables. It was pointed out
that besides removing the ethylene, ethylene removing and absorbing agents
can also remove other organic volatiles from the storage or package atmo-
sphere (Toivonen, 1997). Therefore, at least some effects that were attributed
to the removal of ethylene may in fact be related to the removal of other vola-
tiles that have not been measured or identified. So, there is a strong reason
to evaluate the potential role and significance of volatiles other than ethylene
and their interaction with ethylene in postharvest situations and under differ-
ent storage conditions (Lougheed et al., 1987; Toivonen, 1997). Besides ethyl-
ene, the role of volatiles such as alcohols (mainly ethanol), aldehydes (mainly
acetaldehyde), and methanol has been demonstrated in the ripening of climac-
teric fruits such as tomato (Kelly and Saltveit, 1988; Saltveit and Sharaf, 1992;
McDonald et al., 1996) and apple (Pesis et al., 1994; Pesis, 1995). Besides this,
levels of volatiles are also found to be associated with storage disorders of
apple fruit, such as scald (Huelin and Coggiola, 1968) and internal browning
(Mendoza et al., 2007). The ripening and quality of nonclimacteric fruits such
as grapes, oranges, and strawberries were also influenced by these volatiles
(Saltveit and Ballinger, 1983; Ke and Kader, 1990; Ke et al., 1991).

12.5.2  Flavor and Aroma

In tomato fruit, 17 volatiles have a significant impact on the character-
istic tomato-like aroma (Buttery, 1993). Hexanal, cis-3-hexenal, trans-
3-hexenal, trans-2-hexenal, cis-3-hexenol, 6-methyl-5-hepten-2-one,
β-ionone, 2-­isobutylthiazole, 3-(methylthio)-1-propanol, and 3-(methylthio)-
1-propanal were important in imparting flavor to the fresh red tomato
(Tandon et al., 2000, 2001; Lewinsohn et al., 2001; Baldwin, 2004). Hayata
et  al. (2002) reported that the tomato-like flavor was correlated strongly
with geranylacetone, 2-methylbutanol, 3-methylbutanol, and 6-methyl-5-
hepten-2-one. Distinctive volatile components responsible for aroma and
flavor in some important fruits are presented in Table 12.1.

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 INTERNAL ATMOS P H ERE O F F RUITS

Table 12.1  Distinctive Components of Aroma for Some Fruits

Fruit Compound
Apple ß-Damascenone, butyl hexanoate, isoamyl hexanoate, hexyl
hexanoate, ethylbutanoate, propyl butanoate, hexylbutanoate,
butylacetate, hexanal, 2-hexenal, ethyl 2-methylbutyrate
Banana 2-Hexenal, eugenol, isopentanol, decan-1-ol, 2-phenylethanol,
3-oxy-pentanoic acid, 3-methylbutanoic acid,
3-methylbutylacetate, butanoate, 3-methylbutanoate,
5-methoxyeugenol, eugenol-methyl ether, elemicin
Mango Ethylbutanoate, ethyl-2-butanoate, hexanal, cis-3-hexanal,
trans-2-hexanal, γ-octalactone, γ-dodecalactone, furaneol,
α-pinene, β-pinene, 3-carene, myrcene-limonene, p-cymene,
terpinolene, α-copaene, caryophyllene
Tomato Hexanal, trans-2-hexenal, cis-3-hexenal, cis-3-hexenol, β-ionone,
β-damascenone, 1-penten-3-one, 3-methylbutanal,
3-methylbutanol, 2-isobutylthiazole, 3-(methylthio)-1-propanol,
3-(methylthio)-1-propanal, 1-nitro-phenylethane, trans-2-
heptenal, phenylacetaldehyde, 6-methyl-5-hepten-2-one, methyl
salicylate, geranylacetone
Peach Benzaldehyde, benzyl alcohol, nonanol, linalool, ethyl
hexanoate, 3-methylbutanoate, α-terpineol, γ-hexalactone,
δ-decalactone, γ-undecalactone, δ-dodecalactone, α-pyrone,
6-pentyl-α-pyrone
Orange Geranial, neral acetaldehyde, decanal, octanal, nonanal,
ethylacetate, ethylpropionate, ethylbutanoate, methylbutanoate,
ethyl-2-methylbutanoate, ethyl-3-hydroxy hexanoate, linalool,
α-terpineol, limonene, myrcene, α-pinene, valencene
Lemon Citeral
Grapefruit Acetaldehyde, decanal, ethylacetate, methylbutanoate,
ethylbutanoate, 1-p-menthene-8-thiol, nootkatone, limonene,
naringin
Strawberry Hexanal, cis-3-hexanal, trans-2-hexanal, furaneol, mesifuran,
ethyl hexanoate, ethylbutanoate, methylbutanoate,
ethyl-2-methylpropanoate
Grape Methyl anthranilate, o-aminoacetophenone, furaneol, methyl
furaneol, β-damascenone, β-phenylethanol, butyl alcohol,
hexyl alcohol, hexanal trans-2-hexenal, isoamyl alcohol,
acetaldehyde, isobutyraldehyde, ethylacetate, ethylpropionate,
butylacetate, propylacetate, 2-methylbutanol, linalool,
geraniol, methoxyisobutylpyrazine

(Continued )

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 12.1  Distinctive Components of Aroma for Some Fruits

Fruit Compound
Raspberry 1-(π-Hydroxyphenyl)-3-butanone, α-ionone, β-ionone,
geraniollinalool, benzyl alcohol, ethyl hexanoate,
ethylbutanoate, methylbutanoate, γ-decalactone, 2-heptanone,
cis-3-hexanal, β-damascenone

Sources: Salunkhe, D.K., and Do, J.Y., Critical Reviews in Food Science and
Nutrition 8, 161–190, 1977; Tandon, K.S. et  al., Postharvest Biology
and Technology 20, 261–268, 2000; Tandon, K.S. et al., Proceedings
of the Florida State Horticulture Society 114, 142–144, 2001;
Lewinsohn, E. et al., Plant Physiology 127, 1256–1265, 2001; Baldwin,
E.A., in The Commercial Storage of Fruits, Vegetables and Florist and
Nursery Stocks, Gross, K.C. et al. (eds.), Agriculture Handbook Number
66, Agricultural Research Service, Beltsville, MD, 2004, 18. http://
usna.usda.gov/hb66/023flavor.pdf. Hui, Y.H. (ed.), Handbook of Fruit
and Vegetable Flavors, John Wiley & Sons, Hoboken, NJ, 2010.

The quality of the aroma is related to the concentration and compo-

sition of the volatiles present in the fruit. Negative effects of some of the
above volatiles produced under anaerobic conditions have been perceived
in the quality of aroma (Forney et al., 1991; Hansen et al., 1992), but posi-
tive effects of accumulation of some volatiles were also found to be influ-
enced by volatiles such as acetaldehyde and ethanol (Paz et al., 1981; Pesis
et al., 1986, 1998; Saltveit and Mencarelli, 1988; Frenkel et al., 1995). These
­positive or negative effects were largely found to be dependent on the con-
centrations of ethanol and acetaldehyde in strawberries and persimmon
fruits (Pesis et  al., 1986; Prasad and Stadelbacher, 1974). The sweetness
of tomato fruit was correlated not only with sucrose equivalents and pH,
but also with the volatiles, including cis-3-hexenal, trans-2-hexenal, cis-
3-­hexanol, geranyl-acetone, 2-methylbutanol, 3-methylbutanol trans-2-­
heptenal, 6-methyl-5-hepten-2-one, and 1-nitro-2-phenylethane. Likewise,
sourness was correlated with soluble solids and pH, along with the vola-
tiles, including acetaldehyde, acetone, 2-isobutylthiazole, geranyl-acetone,
β-ionone, hexanal, and ethanol (Saltveit, 2005).

12.5.3  Fruit Decay

Fruit decay means any condition or sign, either physiological or patho-
logical in origin, that makes the fruit unacceptable (Wills and Ku, 2002).
The role of a low O2 -to-CO2 ratio or anaerobic conditions is well known in
determining the overall decay of fruits and vegetables (Banks, 1984). Such
situations were reported to suppress not only the biosynthesis, but also the

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action of ethylene (Kanellis et al., 1989a, 1989b). Ethylene is known to be

involved in the defense against pathogens, as it stimulates the phenylpro-
panoid pathway and synthesis of pathogenesis-related proteins and induces
systemic resistance. These findings explain the higher decay under the
influence of blockage of the stem scar region of tomato fruit, as observed by
Paul et al. (2010b). In tomato fruits, decay was up to 50% or more when the
stem scar portion of fruits was sealed by coconut grease, and it was primar-
ily due to the anaerobic conditions (Calbo et al., 1988; Yang and Shewfelt,
1999). Fruits of highbush blueberry, on the other hand, produce antimicro-
bial volatiles such as trans-2-hexenal that confer resistance to anthracnose
fruit decay (Polashock et al., 2007). Likewise, the preharvest spray treat-
ment of volatile compounds such as ethanol (with calcium) also reduced the
development of gray mold in table grapes (Chervin et al., 2009).

12.6  Ripening Behavior of Some Fruits under

Attached and Detached Conditions

12.6.1 Tomato
Ethylene concentrations were unusually high in attached tomato fruits
(ranging from 2 to 13 µl L –1, depending on cultivars and seasonal condi-
tions), while in detached fruits the values drop to a level of 1.4 µl L –1 on an
average (Sawamura et al., 1978). Detachment of tomato fruit advances the
ripening and reduces the threshold value of endogenous ethylene required
to promote the ripening to a considerably lower level. This supports the
concept of a hypothetical labile ripening inhibitor that is translocated from
vegetative parts of the plant to the fruits, where it antagonizes the action on
ethylene (Sawamura et al., 1978). This concept might be true, but the large
decrease in the levels of ethylene in detached fruits may be explained by
more effective gaseous exchange from the stem scar region of harvested
tomato fruit as a result of detachment of the pedicel. This explanation is
supported by de Vries et al. (1995), who reported that 85%–90% of ethylene
is released by tomato fruit through this stem scar region.
There are various factors that affect the production and release of ethyl-
ene by fruit. In this regard, a theory of ethylene emission was developed and
used as the basis to develop the simulation model called ETHY by Genard
and Gouble (2005). This model was found to be highly sensitive to factors
such as the permeability of the fruit skin; internal concentrations of O2, CO2,
and ACC; changes in fruit growth and temperature; activities of ACC-oxidase
and ACC-synthase; concentration of the ethylene itself; and production sta-
tus of ATP. Similarly, a model has been proposed for gas exchange in pear
fruit (Ho et  al., 2008). The role of surface morphology and the stem scar
region in determining the ripening behavior, rate of respiration, and water

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loss by attached and detached tomato fruits was pointed out, along with a
variety-dependent increase in the density of lenticels with the progress of
ripening (Paul and Srivastava, 2006). Further, it is interesting to note that
the extent of the 1-methylcyclopropene (1-MCP)-mediated delay in ripening
depends on the endogenous level of ethylene in the tomato fruits (Zhang
et al., 2009; Paul et al., 2010a; Paul and Pandey, 2013).

12.6.2 Melons
Several cultivated cultivars and wild ecotypes of melons exhibit a climac-
teric pattern of ripening that is linked with the detachment of the fruit
from its parent plant (Abeles et al., 1992). There are variations in the mag-
nitude and duration of the respiratory rate, depending on the cultivars in
the harvested melon fruits. However, several melon cultivars that appar-
ently do not abscise show no burst in ethylene production and display a
long shelf life. These melon fruits emit little or no ethylene, and therefore
behave like nonclimacteric fruit (Shiomi et al., 1999). It has been reported
that the ­r ipening-associated increase in ethylene production is present,
but the respiratory climacteric is absent during ripening of melon fruit
attached to the plant, leading to the suggestion that climacteric respira-
tion is an artifact of harvest. To address the universality of this phenom-
enon, the ripening behavior in the melon cultivar ‘Charentais’ (Cucumis
melo cv. ‘Reticulatus F1 Alpha’) was investigated. The results, however,
showed that the respiratory climacteric occurs in fruit ripened both on
and off the plant (Hadfield et al., 1995). The sensitivity to ethylene in rela-
tion to the climacteric respiration is affected by detachment of the fruit
(Bower et al., 2002). In an another study, application of ethylene to anti-
sense ACC-oxidase melons stimulated O2 consumption only if they were
detached from the vine, showing that attachment to the plant inhibits the
effect of ethylene on respiration (Pech et al., 2008). This effect of detach-
ment on sensitivity to ethylene is known as the tree factor by posthar-
vest physiologists, as discussed previously in Section 12.6.1. Auxin has
been suggested to be a candidate, but experimental evidence is still lack-
ing (Pech et al., 2008). In muskmelon, the detached fruit also produced
a burst in respiration and ethylene, but attached fruits did not exhibit a
large increase in respiration in spite of the significant increase in the lev-
els of ethylene (Shellie and Saltveit, 1993).

12.6.3  Sweet Pepper

Different ripening patterns exist for attached and detached fruits of sweet
pepper (Saltveit, 1977; Biles et al., 1993; Villavicencio et al., 2001). It was

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reported by Tadesse et al. (1998) that there is an indication that capsicum

fruits of the cultivar ‘Domino’ are climacteric with respect to their internal
ethylene concentration during ripening. But the internal CO2 concentra-
tion, on the other hand, increased only for the fruits undergoing ripening
when attached to the plant, and a steady decline in CO2 was recorded in
fruits that were detached.

12.6.4 Saskatoon
Comparison of respiration and ethylene production rates at nine differ-
ent maturity stages of saskatoon (Amelanchier alnifolia Nutt.) fruits after
their harvest, with the fruits undergoing maturation and ripening on the
plant itself, indicated that the respiratory response was very different in
these two situations. The increase in respiration associated with ripening
was demonstrated only on a whole-fruit basis and only in attached fruits
(Rogiers and Knowles, 1999).
Based on the above information, it could be concluded that cultivar-
dependent variations in the ripening behavior of fruits can also be due to
the differences in their internal gaseous environment. It appears that there
are various factors that contribute differentially to determining the ripen-
ing behavior of attached and detached fruits. Such factors include differ-
ences in the production and release of ethylene, and thereby endogenous
levels of ethylene in the fruit; differences in the sensitivity of fruit to the
endogenous levels of ethylene; differences in the threshold level of ethylene
required to initiate the process of fruit ripening; and overall differences in
the endogenous gaseous microenvironment of fruits. In addition to this,
variability among varieties and cultivars in these factors could also contrib-
ute to the differences in ripening behavior. In view of these reasons, the role
of the above-listed factors was pointed out by Paul et al. (2012) in explaining
the deviations from the classical pattern of climacteric and nonclimacteric
ripening in different fruits.

12.7  Role of Some Gases and Endogenous

Volatiles in Fruit Ripening

12.7.1 Ethylene

12.7.1.1  Role of Ethylene in Ripening of Climacteric Fruits

12.7.1.1.1  Ethylene as the Main Regulator of Ripening in Climacteric Fruits

Ethylene (C2H4) is a naturally produced gaseous plant growth hormone

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many fruits. As stated earlier, this plant hormone plays a major role in
the ­r ipening process of climacteric fruits (Nagata et al., 1995; Klee, 2002;
Bouzayen et al., 2010; Paul et al., 2011; Paul and Pandey, 2013, 2014). The
production of aroma during the ripening of different fruits also depends
strongly on the production and action of ethylene (Golding et  al., 1999;
Rupasinghe et  al., 2000; Lurie et  al., 2002; Flores et  al., 2002; Dandekar
et al., 2004; Pech et al., 2008; Defilippi et al., 2009).

12.7.1.1.2 Regulation of Ethylene Production  Two systems of ethylene

production have been defined in plants (McMurchie et  al., 1972). The
first one is designated as system 1. System 1 operates and functions dur-
ing normal growth and development and in response to various stresses.
System 1 is responsible for the basal level of ethylene production in
­vegetative tissues and unripe fruits. This system is regulated in an auto-
inhibitory manner (Figure 12.1). This means that even the treatment of
exogenous ethylene will not trigger any further synthesis of ethylene. The
second system is system 2, and this operates during floral senescence
and fruit ripening. This system is responsible for the large autoinduc-
tive (autocatalytic) increase in ethylene production during fruit ripening,
especially in the climacteric fruits (Nakatsuka et al., 1998; Inaba, 2007)
(Figure 12.1). High genetic variability in the rate of production of ethylene
has been reported for fruits such as muskmelon, melon, peach, and kiwi-
fruit (Kendall and Ng, 1988; Miccolis and Saltveit, 1991; Klozenbucher
et al., 1994; Xu et al., 1998). Two key enzymes of ethylene biosynthesis,
ACC-synthase and ACC-oxidase, are regulated by the final product of the
reaction, that is, ethylene (Lelievre et al., 1997) (Figure 12.1). Lower eth-
ylene production due to lower activity of ACC-oxidase was assigned as the
cause for the formation of a spongy tissue disorder in ‘Alphonso’ mango
(Nagamani et al., 2010).
Besides the levels of ambient and internal concentrations of ethylene,
the production of ethylene is also governed by abiotic and biotic stresses.
Plants and plant parts are not the only sources of ethylene, but smoke,
exhaust gases, ethylene-releasing chemicals, catalytic production of eth-
ylene from ethanol, and analogs of ethylene (as produced by a variety of
processes within the plant system itself) are also important sources of eth-
ylene or the chemicals that exhibit ethylene-like activities. Such sources of
ethylene and ethylene-like chemicals are common under storage and cold
storage conditions. Different analogs of ethylene are listed in Table  12.2.
These analogs have reduced activity or efficiency, but they can elicit the
same physiological effects as ethylene. So, the presence of analogs of eth-
ylene can also influence the fruit ripening, especially under the storage
conditions.

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Methionine

SAM-synthetase

S-Adenosyl methionine
_ +
ACC-synthase (ACS)

(Auto-induction of ethylene production)

(Auto-inhibition of ethylene production)

1-Aminocyclopropane-1-carboxylic acid
+ +
ACC-oxidase (ACO)

System 2 of ethylene production

System 1 of ethylene production

Ethylene
response is suppressed in

1-MCP _
presence of 1-MCP
Ethylene-mediated

Perception of ethylene by its binding

to ethylene binding receptor

Response
(Ripening and senescence, etc.)

Figure 12.1  Pathway of ethylene biosynthesis in plants showing autoinhi-

bition (inhibiting its own production) and autoinduction (inducing its own
production) of ethylene. These systems are referred to as systems 1 and 2
of ethylene production, respectively. In system 1, ethylene inhibits its own
production by inhibiting ( − ) ACS expression or activity. It may be noted that
the ACO activity is enhanced during system 1, but due to the absence of any
enhancement in the activity of ACS, there is no autoinduction. In system 2,
ethylene induces more of its own production by stimulating ( + ), the expres-
sion or activity of both enzymes (ACS and ACO) simultaneously, leading to
a burst in ethylene production. (Adapted and modified from Paul, V. et al.,
Journal of Food Science and Technology 49, 1–21, 2012.)

12.7.1.2  Role of Ethylene in Ripening of

Some Nonclimacteric Fruits

12.7.1.2.1 Strawberry  Strawberry is considered to be a typical example

of nonclimacteric fruit, and it has served as a model system for this cat-
egory of fruits (Giovannoni, 2001). Application of the highly sensitive
technique of laser photoacoustic spectroscopy, coupled with the specific

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Table 12.2  Relative Activity of Ethylene and Its Analogs in Pea Straight
Growth Bioassay Test
ppm (µl L–1) in Gas Phase for
Gases Formula Half-Maximum Activity
Ethylene C2H4 0.1
Propylene C3H6 10
Vinyl chloride C2H3Cl 140
Carbon monoxide CO 270
Acetylene C2H2 280
Vinyl fluoride C2H3F 430
Methylacetylene (propyne) C3H4 800
Vinyl bromide C2H3Br 1,600
Allene (propadiene) C3H4 2,900
Vinyl methyl ether C3H6O 10,000
Ethylacetylene (1-butyne) C4H6 11,000
1-Butene C4H8 27,000
Vinyl ethyl ether C4H8O 30,000
Carbon dioxide CO2 30,000
1,3-Butadiene C4H6 500,000
Sources: Abeles, F.B. et al., Ethylene in Plant Biology, 2nd ed., vol. 15, Academic
Press, San Diego, 1992; Burg, S.P., and Burg, E.A., Physiologia Plantarum
18, 870–886, 1965; Abeles, F.B., and Gahagan, H.E., Plant Physiology
43, 1255–1258, 1968.

apparatus to determine in planta ethylene production in fruits and ­flowers

of strawberry during development and ripening, revealed an increase
in ethylene production and a concomitant rise in respiration rate in red
strawberry fruits (Iannetta et  al., 2006). Treating strawberry fruit with
the ethylene action inhibitor (silver thiosulfate) showed that this increase
in ethylene production was under the control of a positive feedback mech-
anism in ripe fruits, thereby suggesting that a form of autoinduced eth-
ylene production is operational during ripening in strawberry (Iannetta
et al., 2006). However, the timing of this ripening-related increase in eth-
ylene production in strawberry was distinct from the patterns of ethylene
production typically found in the ripening of climacteric fruits such as
tomato (Iannetta et al., 2006). Trainotti et al. (2005) reported an increase
in the synthesis of ethylene receptors concomitant with the increased
synthesis of ethylene in strawberries. The receptors mostly expressed
in ripening strawberries are type II ones (with a degenerate histidine–
kinase domain). According to Cancel and Larsen (2002), the degenerate

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histidine–kinase domain of type II receptors leads to a weaker affinity for

the amino-terminal domain of CTR1 than that of type I receptors. So, even
the little ethylene produced by strawberries (Abeles and Takeda, 1990;
Perkins-Veazie et  al., 1996; Iannetta et  al., 2000) might be sufficient to
trigger ripening-related physiological responses. This means that CTR1
might be released by type II receptors, such as FaEtr2, by lower amounts
of ethylene. In addition to this, an increase in the expression of FaEtr2 (a
type II receptor homolog that is closely related to the tomato LeETR4)
was also found to be associated with ripening in strawberry (Trainotti
et al., 2005).

12.7.1.2.2 Citrus  Studies on citrus, a nonclimacteric fruit, showed that

ethylene (endogenous as well as exogenous) plays a role in regulating the
ripening of the peel in citrus fruit (Goldschmidt, 1997). Ethylene stimu-
lates respiration, chlorophyll breakdown, and carotenoid accumulation
(Goldschmidt, 1997; Purvis and Barmore, 1981; Katz et al., 2004). Further
studies have shown that some genes (including chlorophyllase) were regu-
lated by ethylene in citrus fruit (Alonso et al., 1995; Jacob-Wilk et al., 1999).
Earlier, it was reported that there is a significant rise in ethylene evolu-
tion and respiration in young citrus fruits, and later Eaks (1970) proposed
this phenomenon as “pseudo-climacteric.” According to McMurchie et al.
(1972), citrus (and other nonclimacteric fruits) lack a ripening-associated
autocatalytic rise in ethylene production. But, at the same time, they also
suggested that this does not rule out a role for the small but detectable
amount of endogenous ethylene in the regulation of the ripening process. In
a review article, Goldschmidt (1997) pointed out the potential significance
of even the lowest levels of ethylene present in nonclimacteric fruits in regu-
lation of the ripening process. Induction of ripening of citrus fruit peel by
ethylene was also found to be opposed by gibberellins and cytokinins, and
this was possibly due to the reduction in the tissue’s sensitivity to ethyl-
ene. There is autoinduced ethylene production in citrus fruit (Katz et al.,
2004). The harvested immature fruits produce high levels of ethylene, and
this can be further stimulated by exogenous treatment with ethylene or
propylene and inhibited by 1-MCP. This indicates the autoinductive nature
of ethylene production, but with altered timing, in comparison to similar
events in climacteric fruit (Katz et  al., 2004). Likewise, the developmen-
tal regulation of transition from system 1 to system 2 of ethylene produc-
tion in climacteric fruit and the transition from system 2 behavior of young
fruitlets of citrus to system 1 behavior by mature fruit were also suggested
to be under the developmental regulation (Katz et  al., 2004). Changes in
the sensitivity to ethylene with development or ripening have already been
pointed out by many workers in different fruits (Goldschmidt et al., 1997;
Katz et al., 2004; Van Loon et al., 2006; Pech et al., 2008; Yoo et al., 2009;
Johnston et al., 2009).

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12.7.1.2.3  Grape Berry  Ethylene is now reported to influence multiple

aspects of ripening in grape, although grape is regarded as an important
example of a nonclimacteric fruit. Similar to strawberry, a small but sig-
nificant increase in ethylene synthesis (at the onset of ripening) has been
detected in grape berries also. Chervin et  al. (2004) demonstrated the
presence of a transient peak of ethylene production in grapes just prior
to the onset of ripening. Experiments with 1-MCP (action inhibitor of
ethylene) indicated that ethylene is required for the onset of accumula-
tion of anthocyanins, fruit swelling, and the decrease in acidity associ-
ated with the ripening of grape berries (Chervin et al., 2004). The results
therefore indicate that tissues of grape berry have a fully functional path-
way for ethylene synthesis, and this pathway is activated and the percep-
tion of ethylene is critical just before the veraison stage (Chervin et al.,
2004). Ethylene treatment to grape berries stimulated an increase in the
­production of anthocyanins, along with an increase in the four transcripts
encoding enzymes (involved in synthesis of anthocyanins) (El-Kereamy
et al., 2003). It is observed that low doses of ethylene application increase
the berry diameter at the inception of the veraison stage d ­ uring ripening
(Chervin et al., 2008).

12.7.1.2.4 Melons  This group of fruits comprises not only climacteric

but also nonclimacteric genotypes. More understanding is now achieved
by making use of near-isogenic lines (NILs) of melon, where various
volatiles such as esters, aldehydes, isobutylacetate, benzylacetate, and
pentanal allowed the climacteric NILs to be distinguished from the non-
climacteric NILs at harvest using univariate and multivariate analyses
(Obando et al., 2007; Obando-Ulloa et al., 2008a, 2008b, 2009). Wechter
et al. (2008) performed ethylene bioassays in a closely related watermelon
genotype (having similar phenotypes) at different (green, white, pink,
and red) stages. This study showed that levels of ethylene were highest
during the green stage of fruit development, and this was followed by a
decrease in its levels during the white and pink stages of development.
The higher level of ethylene production during fruit development in
watermelon thereby supports a role for ethylene in development, besides
its role in ripening of this fruit.

12.7.1.2.5 Cucumber  Cucumber fruits are classified as nonclimacteric

(Biale and Young, 1981); however, a climacteric-like increase in posthar-
vest ethylene production has been reported in several cultivars of cucum-
ber. It was noticed that ethylene production patterns were more closely
associated with decay than climacteric respiratory patterns or ripening-
like physical changes (e.g., color or softening) (Saltveit and McFeeters,
1980). A recent study by Hurr et  al. (2009) investigated the postharvest

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storage and ethylene responses of cucumber fruit at defined developmental

stages based on growth and surface color parameters. The results showed
that ethylene increased respiration in fruit of all stages of development,
but ethylene ­production was detectable only in decaying fruit. The data
also showed that the postharvest response of cucumber fruit to ethylene
is highly dependent on the developmental stage because only older fruits
exhibit climacteric-like responses, while younger fruits exhibit only tissue
water soaking and general fruit deterioration.

12.7.1.2.6 Pepper  Capsicum fruits are classified as nonclimacteric

based on the patterns of CO2 and ethylene production (Biles et al., 1993).
However, some hot pepper cultivars are climacteric (Gross et  al., 1986),
indicating that classification of capsicum as nonclimacteric may be incon-
clusive. Some cultivars of pepper seem to be ethylene insensitive, while
continuous treatment with exogenous ethylene has been shown to acceler-
ate ripening in others, along with enhanced expression of ripening ­specific
genes (Harpster et al., 1997). Villavicencio et al. (2001), on the other hand,
reported that all cultivars seem to exhibit intermediate levels of ethylene
and CO2 evolution during ripening, and therefore peppers appear to fall
between the typical characteristics of climacteric and nonclimacteric
behavior of fruit ripening.

12.7.2  Oxygen and Carbon Dioxide

12.7.2.1  Low Oxygen

In general, levels of oxygen (O2) show a decreasing trend from the outer
to the inner parts of the fruit (Figure 12.2). The difference in the depletion
of internal O2 levels in different kinds of fruit under MA conditions was
observed (Sornsrivichai et  al., 1998). Yip et  al. (1988) claimed that 50%
reduction in ethylene production could be obtained at a 1% level of O2 . This
is primarily because O2 is itself a substrate for the reaction catalyzed by
the enzyme ACC-oxidase, as well as the action of ethylene in fruits, includ-
ing tomato (Burg and Burg, 1965b) (Figure 12.3). Fruits show reduction
in respiration with lowering of O2 in the surrounding atmosphere, and at
a specific reduced level of O2 , there is induction of anaerobic respiration.
This leads to a fast breakdown of sugars and is referred to as the Pasteur
effect (Boersig et  al., 1988; Kader, 1986). The Pasteur effect has practi-
cal importance in the modified atmosphere storage of fruits (Weichmann,
1986). Oxygen concentrations must be managed so that aerobic respiration
is minimized, but anaerobic respiration (which leads to the fast breakdown
of sugars) is avoided (Kader, 1986; Weichmann, 1986; Kubo et al., 1996).

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Epidermis
Mesocarp

Seed cavity

7 22
6 21
5 20
% Carbon dioxide

4 19

% Oxygen
3 18
2 17
1 16
0
Distance

Figure 12.2  Cross section passing through a fruit showing how the con-
centration of O2 and CO2 can vary within different tissues of the fruit due
to respiration and external and internal barriers to gas diffusion. (From
Kader, A.A., and Saltveit, M.E., in Postharvest Physiology and Pathology of
Vegetables, Bartz, J.A., and Brecht, J.K. (eds.), New York, Marcel Dekker,
2003, 229–246.)

It was reported by Saltveit (2003) that the optimum level of O2 concentra-

tion needed to maintain the aerobic respiration is not only different for dis-
similar commodities, but also shifts for a given commodity over a period of
time during storage. In tomato fruits, low O2 not only caused an increase in
the production of ethanol and acetaldehyde, but also delayed the ripening
in comparison to the control (Klieber et al., 1996). In bulky and dense stor-
age organs (such as apple fruit), the internal O2 concentration may fall to
low levels of 8%–10% near the surface, and even to very low levels of 2%–5%
in the center. Such conditions may enhance anaerobic respiration and trig-
ger the accumulation of acetaldehyde and ethanol (Rolletscheck et  al.,
2002). In tomato and pear fruits, hypoxia can also result in an increase
in activity of pyruvate decarboxylase (PDC) and ADH (Chen and Chase,
1993). Below a certain level of O2 , the rise in CO2 production indicates a

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L-Methionine

SAM-synthetase

SAM
Low concentration of
+
SA/MSA/JAs, water stress
NO ACC-synthase
– > CO2, ethanol/acetaldehyde,
high concentration of SA/MSA/JAs
ACC
O2, < CO2, low concentration of
+
NO ACC-oxidase JAs, water stress

– > CO2, ethanol/acetaldehyde,

high concentration of SA/MSA/JAs

Ethylene, low
+
Auto-induced concentration of SA/MSA
NO Ethylene ethylene
production – > CO2, ethanol/acetaldehyde,
NO, high concentration of
SA/MSA/JAs
NO + Ethylene
< O2/CO2, hypoxia, anoxia, ethanol/
–
acetaldehyde (concentration dependent)

Response (ripening and senescence, etc.)

Figure 12.3  Different endogenous volatiles and other factors that play a
regulatory role in determining the production and response of ethylene. The
symbols + , − , , >, and < indicate inducers, suppressors, higher, lower,
and possible interactions, respectively. SAM, S-adenosyl-L-methionine; ACC,
1-amino-cyclopropane-1-carboxylic acid; SA, salicylic acid; MSA, methyl
salicylic acid; JAs, jasmonates; NO, nitric oxide.

switch to fermentative metabolism. This O2 level has been called the anaer-
obic compensation point (ACP) (Leshuk and Saltveit, 1990). The ACP may
vary for different fruits and for the same fruit at different maturities and at
different storage temperatures, besides being affected by different variet-
ies of a given fruit (Gran and Beaudry, 1993; Kubo et  al., 1996; Borisjuk
et al., 2007).
The susceptibility of apple fruit to low O2 injury in CA storage was
found to be positively correlated with the resistance of the fruit to the dif-
fusion of gases. For a given strain or cultivar, resistance to gas diffusion
was found to be affected by the fruit’s maturity, duration of storage, and
whole-fruit volume (Park et  al., 1993). It was noticed that the ‘Marshall’
strain of McIntosh apple had higher resistance, and thereby it showed more
susceptibility to low O2 injury, as this strain accumulated 10 times more

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

ethanol than ‘Rogers’ (another strain of McIntosh apple) (Park et al., 1993).
This demonstrates the extent of varietal variability among the fruits for
response toward their internal atmosphere, especially for the lower con-
centrations of O2 .
Modified atmospheres with low concentrations of O2 can slow down
the deterioration of fruits by decreasing respiration, ethylene production,
and tissue sensitivity to the ethylene (Kader et  al., 1989). The extent of
decrease in ethylene production therefore depends not only on the O2 con-
centration that is present in the fruit’s internal atmosphere, but also on the
sensitivity of the ethylene production system under a prevailing concen-
tration of O2. Sanders and de Wild (2003) reported a lower partial pres-
sure of O2 (lower than the external partial pressure of O2) due to the rapid
consumption of O2 by the tomato fruit and higher resistance of fruit to the
diffusion of O2. Reduced O2 or elevated CO2 decreased the respiration rate
(Smith et al., 1987). In bulky storage organs such as fruits (where the length
of the diffusion path may be considerable), hypoxia conditions have been
demonstrated (Ho et al., 2008). So, low O2 stress may occur within the fruit
as it grows, and the resistance to the entry of O2 from the atmosphere into
the fruit (via the diffusion process through the skin and thickened cell lay-
ers of the cortex) becomes significant. In avocado fruit, synthesis of cellu-
lase and polygalacturonase was found to be directly related to the levels of
O2 (Knee, 1982; Kanellis et al., 1991).

12.7.2.2  High Carbon Dioxide

There is a decreasing gradient in CO2 concentrations from the inner parts
of the fruit to the surface, which is reverse to the gradients observed for
O2 (Figure 12.2). High CO2 was reported to reduce the activity or syn-
thesis of various enzymes of respiratory metabolism (Lange and Kader,
1997), including oxidative phosphorylation (Shipway and Bramlage,
1973). Studies on respiration and the factors influencing the respiration
are important because the potential shelf life of harvested plant parts
(including fruits) was found to be closely related to the rate of respira-
tion of the plant part (Varoquaux and Ozdemir, 2005). Bufler (1984) and
Mathooko et al. (1995) reported that CO2 at high concentrations competi-
tively inhibits the effects of ethylene by preventing the autoinduction of
ACC-synthase, as shown in Figure 12.3. As per Burg and Burg (1967), the
amount of CO2 in the intercellular spaces of fruits at the preclimacteric
stage is low, but may approach higher levels of around 10% during ripening
and the postclimacteric phase. This higher endogenous level of CO2 prob-
ably raises the threshold concentration of ethylene to higher levels for
its action in fruits. It has been demonstrated that elevated CO2 (5%–20%)
inhibits ethylene production in climacteric fruits by inhibiting activities of

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 INTERNAL ATMOS P H ERE O F F RUITS

ACC-synthase and ACC-oxidase (Mathooko et  al., 1995; Chavez-Franco

and Kader, 1993).
Work carried out by Chaves and Tomas (1984) suggested that CO2
interferes with ethylene metabolism through a mass action effect. Besides
this, displacing ethylene from its receptor site has also been proposed
(Yang and Hoffman, 1984). Using a continuous flow through a gas system,
it has been demonstrated that 20% CO2 markedly decreases ethylene bio-
synthesis in ripening peaches by delaying and suppressing ACC-synthase
at the transcriptional level; however, recovery occurs upon withdrawal of
CO2 (Mathooko et al., 2001). About a 1% level of CO2 stimulatory effect on
the production of ethylene could be due to a balance between its stimu-
latory effect on the activity of ACC-oxidase and inhibitory effect on the
activity of ACC-synthase, wherein the contribution of the former is more
significant (Mathooko, 1996). All elevated levels of CO2 inhibit the activity
of ACC-synthase, while the activity of ACC-oxidase is differentially regu-
lated by CO2 (it is stimulated at low CO2 levels but inhibited at high CO2
levels) (Mathooko, 1996) (Figure 12.3). Tolerance of a commodity to ele-
vated levels of CO2 depends on its physiological condition, maturity status,
CO2 concentration within the tissue, duration of exposure, internal O2 con-
centration, and storage temperature (Zagory and Kader, 1988). Tian et al.
(1994) proposed that the mechanism of CO2 stimulation of ACC-oxidase
may be direct and probably through interaction with a nonsubstrate bind-
ing site on ACC-oxidase. They further stated that CO2 might combine
reversibly with an ACC-oxidase–ACC complex to increase Vmax (the maxi-
mum velocity or rate at which the enzyme catalyzes a reaction, it happens
when all enzyme active sites are saturated with substrate) of the reaction.

12.7.2.3  Ratio of O2 to CO2

As early as 1936, Wardlaw and Leonard reported that respiratory climac-
teric is an anaerobic type of respiratory shift. Later on, climacteric rise was
considered a type of anaerobiosis because fruits naturally ripen from the
inside outward (Leonard and Wardlaw, 1941). Diagrammatic representa-
tion of fruit in Figure 12.2 shows how the concentrations of O2 and CO2 vary
within the fruit. Respiration by the fruit tissues and barriers of the diffusion
and exchange of gases as posed by the anatomical, morphological, physical,
and biochemical components (present either on the surface or inside the
fruit) are responsible for the gradual lowering of the O2 -to-CO2 ratio from
the outside to the inside of the fruit.
As described above and presented in Figure 12.3, conditions such as
hypoxia or a low O2 -to-CO2 ratio reduce the synthesis as well as the action of
ethylene (Gorny and Kader, 1996). Hypoxia was also reported to reduce the
expression of genes involved in the maturation process, which are regulated

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

by ethylene (Kanellis et al., 1993). Production of volatiles in fruits has been

shown to be altered in high CO2 or low O2 conditions (Larsen, 1994). Besides
this, high CO2 or low O2 within the atmosphere of the fruit, or both, can induce
anaerobic metabolism, resulting in enhanced accumulation of ethanol and
acetaldehyde (Kader, 1987). Ethanol and acetaldehyde were in fact reported
to delay the ripening of tomato fruit (Beaulieu et al., 1997). CO2 at the level
of 10 kPa, in combination with 6 kPa of O2, was suggested to be suitable for
cold storage for late and early harvested grapes for up to 12 and 4 weeks,
respectively, as this limits the losses due to gray mold (Crisosto et al., 2002).
The biological variation in the apparent diffusivity of gases in tissue
was related to the natural and random distribution of cells and pores in
the cortex tissue (Ho et  al., 2009). Model-based in silico analysis for the
exchange of O2 and CO2 in pear fruit showed that O2 exchange takes place
mainly through the intercellular spaces and the cell wall network and mar-
ginally through the intracellular liquid (cytoplasm). On the other hand, CO2
exchange occurs at similar rates through each of these phases (Ho et al.,
2009). In this way, anatomical features can be considered responsible for
the already existing differences in the tolerance to reduced O2 or elevated
CO2 levels among various fruits and vegetables. The promotive or inhibi-
tory effects of O2 and CO2 levels, the O2 -to-CO2 ratio, and conditions such as
hypoxia, anoxia, and various volatile metabolites are summarized for their
effect on different steps of ethylene biosynthesis and ethylene response (or
action) in Figure 12.3.

12.7.3  Ethanol and Acetaldehyde

Ethanol production generally results from low concentrations of O2, which
can be caused by either reduced levels of external O2 or enhanced resis-
tance to the diffusion of O2 into the plant parts and fruits (Jackson et al.,
1982). Fruits undergoing the developmental and ripening process exhibit
changes in the levels of O2 and CO2 inside them. These changes are usu-
ally in a direction that leads to a net reduction in the O2 -to-CO2 ratio within
the fruit and results in the accumulation of ethanol (Bufler and Bangerth,
1982). Loss of ethanol from fruit occurs predominantly by the evaporation
process, and this is mainly determined by the degree of diffusion resistance
posed by the fruit in view of its surface and anatomical features, as already
described above. Effective ethanol concentrations for delaying the ripening
were, however, found to be similar to those of endogenously built-up levels
of ethanol, as observed in the following conditions: during ripening of fruit
(Bufler and Bangerth, 1982), during anoxia (Jackson et  al., 1982), over a
few days of anaerobiosis (Kelly and Saltveit, 1988), under stress (Kimmerer
and Kozlowski, 1982), and with fruits exposed directly to ethanol vapors
(Kelly and Saltveit, 1988). Ethanol appears to inhibit not only the synthesis

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 INTERNAL ATMOS P H ERE O F F RUITS

of ethylene, but its action as well (Saltveit and Mencarelli, 1988; Ritenour
et al., 1997; Pesis, 2005; Asoda et al., 2009). Ethanol was also reported to
delay ripening as well as the production of CO2, loss of chlorophyll, and
synthesis of lycopene (Kelly and Saltveit, 1988; Yang and Shewfelt, 1999;
Podd et al., 2002).
It has been proposed that the ethanol-mediated delay in ripening is
basically caused by acetaldehyde. Acetaldehyde is produced by conver-
sion of ethanol to acetaldehyde via the reversible reaction catalyzed by the
enzyme ADH (Beaulieu et  al., 1997; Podd et  al., 2002). Both climacteric
and nonclimacteric fruits produce a lot of acetaldehyde and ethanol (Pesis,
2005). For a given fruit, genetic variability was also seen in the levels of
production of acetaldehyde and ethanol and in the ability to survive under
anaerobiosis (Pesis, 2005). Acetaldehyde inhibits the formation of ethylene
by preventing the action of ACC-synthase and action and synthesis of ACC-
oxidase (Pesis et al., 1998; Podd et al., 1998). Exogenous ethanol application
resulted in a marked increase in acetaldehyde levels, and this inhibited the
ethylene production and ripening of tomato fruits (Pesis and Marinansky,
1993). It was therefore suggested that acetaldehyde is the causal agent for
ethanol-induced inhibition of fruit ripening (Beaulieu et al., 1997). In light
of the above findings, it was concluded by Pesis (2005) that ethanol and
acetaldehyde are natural compounds that are essential in governing the
process of fruit ripening. These compounds are also associated with aroma
production and the removal of astringency. Various sites where acetalde-
hyde and ethanol can regulate the production and response of ethylene are
presented in Figure 12.3.

12.7.4  Water Vapors and Water Status in Fruit

Fruit maintains vascular continuity with the mother plant and receives
water as long as it remains attached to the plant. But, once detached, the
fruit has no renewable source of water to compensate for the water that
is being lost through transpiration. Detached fruit therefore experience
water stress. Water loss (transpiration) from freshly harvested fruit results
in loss of salable weight, appearance (wilting and shriveling), textural qual-
ity (softening, flaccidity, limpness, and crispness), juiciness, and nutritional
quality (Kader and Barrett, 1996).
The transpiration rate of harvested fruit depends mainly on the rate
of cooling of fruit after its harvest, the structure and condition of the fruit
surface, the surface-to-volume ratio of the fruit, the relative humidity and
temperature during storage, the air movement and circulation, and the
atmospheric pressure in the storage environment (Gamage and Rahman,
1999). The main sites of transpiration in the plant and its parts are the sto-
mata, epidermal cells, lenticels, trichomes (hairs), stem scar, hydathodes,

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

and cuticular cracks (Ben-Yehoshua, 1987). The surface characteristics,

such as number of stomata on the epidermis, type of surface, tissue under-
lying the skin and structure, and thickness and chemical composition of
the wax and cuticle, play a role in determining the water loss from fruit,
and these features vary greatly among the fruits and also with the develop-
mental stages for a given fruit. The stem scar region is an important path-
way for water loss in tomato fruit. In apple, lenticels account for up to 21%
of the transpiration (Maquire et  al., 2001). Complete coating of fruit was
found to retard gaseous exchange by plugging the stomatal pores of citrus
(Ben-Yehoshua et al., 1985). Such coatings reduced weight loss up to 20%
in mandarin (Lawes and Prasad, 1999), mango (Baldwin et al., 1999), and
pear (Amarante et  al., 2001). Variability in peel permeance, weight loss,
and internal atmosphere was recorded in different lines of mandarin fruit
(Lawes and Prasad, 1999).
It was reported by Grierson and Wardowski (1978) that water loss
after harvesting causes reduction in fruit mass, and it may also induce
senescence. Water deficit in plant tissues also stimulates ethylene produc-
tion (Figure 12.3), and as a consequence, there is an increase in the respira-
tion of tissues (Yang and Pratt, 1978). Increased ethylene production was
found when fruit was detached and also after having experienced water
stress and deficit during its development (Gelly et al., 2003). The level of
transcription for the genes of ACC-synthase and ACC-oxidase responded
positively to the water deficit in a tissue-specific and coordinated manner
(Nakano et  al., 2002). This resulted in enhanced activities of these two
enzymes under water stress (Figure 12.3). Several authors found that water
stress decreases the preclimacteric life of many fruits, such as banana, avo-
cado, pear, and plantain (Littmann, 1972; Adato and Gazit, 1974; George
et al., 1982). It was found by Finger et al. (1995) that an increase in respira-
tion and the evolution of ethylene was significantly higher when the fruits
reached 5% of loss in terms of fresh mass. The increase in respiration was
70%, and ethylene production was 50% more for such fruits when compared
to the control fruits in banana. The results thereby confirmed that water
stress (after the harvest of fruit) might affect the shelf life, depending on
the intensity of the water stress.
Water content alone is not sufficient to describe the water status of
different parts of fruits, its movement in fruits, and the associated physi-
ological and biochemical changes occurring in the fruits (Nguyen et  al.,
2004). The moisture distribution (water content) is related to the water sta-
tus (water potential). So to understand water movement in fresh fruit, a
detailed modeling of cellular structural properties and intercellular spaces
is needed (Nguyen et  al., 2004). Water activity (aw) is considered a more
reliable parameter of water status, as it includes the contribution of the
pressure forces in the tissue. Water activity is important in the determi-
nation of the stability criteria of foodstuffs in terms of microbial growth,

386
 INTERNAL ATMOS P H ERE O F F RUITS

browning, lipid oxidation, ripening, ripening-related changes, and shelf life

(Joyce et al., 2002).

12.7.5  Salicylic Acid and Methyl Salicylate

Salicylic acid (SA) and its volatile derivatives, such as methyl salicylate
(MeSA), were characterized as inhibitors of ethylene biosynthesis (Leslie
and Romani, 1986). At low concentrations, MeSA appears to enhance the
ripening processes in tomato (Ding et  al., 2002). Dual effects of MeSA
on ethylene metabolism (inhibitory and promotive) were suggested to
be dependent upon the dose and developmental stage of fruits (Ding and
Wang, 2003). It was clearly demonstrated that low concentrations of MeSA
have the potential to upregulate ethylene biosynthesis by increasing the
expression of ACC-synthase genes LE-ACS2 and LE-ACS4 (responsible for
autoinduced production of ethylene during ripening of tomato fruit) and
eliminating the transcription of LE-ACS6 (genes of ACC-synthase respon-
sible for the basal level of ethylene production in tomato fruit) (Ding and
Wang, 2003). On the other hand, high levels of MeSA in tomato fruit keep
the ACC-synthase and ACC-oxidase genes repressed and thus inhibit the
timely production of ethylene during ripening (Ding and Wang, 2003). In
this way, the endogenous concentration of MeSA could be critical in deter-
mining ethylene metabolism (Figure 12.3), depending on the stage of
the fruit.

12.7.6  Jasmonic Acid and Jasmonates

Jasmonic acid, its volatile ester (i.e., methyl jasmonate [MJ]), and other
derivatives are collectively known as jasmonates (JAs). JAs play a role dur-
ing developmental processes, including pollen development, seed germina-
tion, root growth, and fruit growth and ripening (Pena-Cortes et al., 2005;
Wasternack, 2007). Multiple interferences and interactions between JAs and
ethylene signaling pathways were studied by analyzing Arabidopsis mutants
(Devoto and Turner, 2005). Furthermore, JAs were reported to regulate the
early steps of climacteric fruit ripening by stimulating ethylene biosynthe-
sis (Kondo et al., 2000). The application of exogenous JAs stimulates ethyl-
ene production and color change in tomato (Saniewski and Czapski, 1985).
It was found that activities of ACC-synthase and ACC-oxidase were stimu-
lated by JAs in a concentration range of 1–100 µM. But, continuous expo-
sure at a higher dose, that is, 1000 µM, inhibited activity of both of the above
enzymes (Figure 12.3) (Fan et al., 1998). The convergence of ethylene and
jasmonate pathways at the transcriptional level of ethylene response factor
1 (ERF1) was proposed (Lorenzo et al., 2003). MJ induced the transcription
of ACC-oxidase in tomato fruit (Imanishi et al., 2005). The mRNA levels of

387
P OST H AR V EST RI P ENING P H Y SIOLOG Y

81 genes were found to show elevation (by more than threefold) in response
to MJ in wild-type tomato in comparison to nor mutant (Imanishi et  al.,
2005). The results indicate a strong interaction of jasmonate and ethylene,
and in this way, the endogenous status of jasmonate could regulate the rip-
ening and ripening-related changes. In a study by Ziosi et al. (2007), exog-
enous application of JAs led to alterations in ethylene biosynthesis, ethylene
perception, and polyamine accumulation.

12.7.7  Nitric Oxide

It was shown by Leshem et al. (1998) that exogenous NO extends the post-
harvest life and delays senescence in fruits, vegetables, and flowers. Later,
it was made clear that NO is a natural senescence-delaying plant growth
substance that acts by downregulating ethylene production (Leshem et al.,
2000). NO affects ethylene production through its direct regulatory effect
on ACC-synthase or ACC-oxidase enzymes (Figure 12.3), as well as on
their genes (Wills et  al., 2000). Application of NO to tomato delayed the
burst in ethylene production and color development at the green mature
and breaker stages of fruits, but not at the pink and full red stages (Eum
et  al., 2009). The results therefore implicate that NO might control the
postharvest metabolism of fruits, depending on its dose and the status
of the commodity that is being treated. Borisjuk et al. (2007) proposed a
key role of NO in the immediate sensing and balancing of the O2 status
in plants. Depletion in the level of O2 and conditions such as hypoxia or
anoxia were already reported to prevail in fruit undergoing maturation and
ripening (Franck et al., 2007; Ho et al., 2008; Lammertyn et al., 2003). So,
NO-mediated sensing of O2 and the balancing of its levels in fruits cannot
be ruled out. Wills et al. (2000), Leshem et al. (2000), and Eum et al. (2009)
also reported that NO affects the ethylene production through its direct
regulatory effect on the enzymes of ethylene biosynthesis (ACC-synthase
and ACC-oxidase) and also on their genes during the ripening of tomato.
The regulatory and interactive linkages of NO with the biosynthesis and
response of ethylene are presented in Figure 12.3.

12.8  Internal Atmosphere of Fruits: Practical

Implication in Ripening and Storability
The permeability of different gases and volatiles is governed by the dif-
fusivity of these compounds in air, water, and across the cellular micro-
structures of fruit. The microstructures of the internal and surface tissues
of fruit include their cellular details, porosity (up to the microlevel), and
3-D organization of cells. Fick’s law, as such, cannot be applied directly to

388
 INTERNAL ATMOS P H ERE O F F RUITS

predict the gaseous concentration in fruits because fruit tissue does not rep-
resent a continuum. Permeability therefore changes in a dynamic way with
metabolic activities of tissues, developmental stage, maturity, and storage
time. Besides this, the external factors, such as the temperature, relative
humidity, and gaseous composition of the immediate storage environment,
also affect the permeability within and across the fruit. Several researchers
have described different geometrical models with mathematical equations
that also take into the consideration the tissue microstructures, besides
other factors (Verboven et  al., 2008; Ho et  al., 2008, 2009, 2010; Steindel
et al., 2005).
Developments in the field of plant volatiles and their roles showed that
the quality and quantity of volatiles in plants and plant parts are linked to
various types of abiotic and biotic stresses (Steindel et al., 2005; Karl et al.,
2008; Loreto and Schnitzler, 2010). Some specific examples are hexanal,
trans-2-hexenal, and hexylacetate, which improve the safety of freshly sliced
apple (Lanciotti et al., 2003); trans-2-hexenal, which confers resistance to
anthracnose fruit decay in the highbush blueberry (Polashock et al., 2007);
hexanal, which reduces infection of tomatoes by Botrytis cinerea (Utto
et al., 2008); α-farnesene, which increases the susceptibility of apple fruits
to scald disorder (Watkins et al., 1993); different endogenous volatiles (alde-
hydes, alcohols, and esters) and trans-2-hexenal, which exhibit antifungal
properties against Colletotrichum acutatum, which causes anthracnose in
strawberry fruits (Arroyo et al., 2007); several terpenoids (sesquiterpenes
such as zingiberene and curcumene and monoterpenes such as p-cymene,
α-terpinene, and α-phellandrene), which strongly repel whiteflies that
infest tomatoes (Bleeker et  al., 2009); and isoprene, which is reported to
provide thermotolerance in Arabidopsis (Sasaki et al., 2007)—however, its
role in fruits is yet to be studied. Strawberries treated with MJ alone or in
combination with ethanol showed higher antioxidant capacity, total pheno-
lics, and anthocyanins, along with longer postharvest life and better quality
and aromatic properties (Ayala-Zavala et al., 2005). These examples show
that at least some of the volatiles in the internal atmosphere of fruit can
provide protection against biotic and abiotic stresses during ripening and
storage and reflect potential practical applications in the overall posthar-
vest management of fruits.
Today, efforts are being made for the more effective use of some gases,
such as carbon monoxide (CO), nitrous oxide (N2O), nitrogen (N2), sulfur
dioxide (SO2), and chlorine dioxide (ClO2), in the storage environment
to reduce the microbial, insect, and pest infestation. Besides this, ozone
(O3) gas is also being exploited in delaying the ripening and improving the
quality of the stored fruits (Scully and Horsham, 2008; Hoehn et al., 2009;
Zambre et al., 2010; Rodoni et al., 2010). Ozone-treated fruits are reported
to show a delay in ripening and decay, the proper maintenance of quality,
suppression in microbial contamination, and a reduction of the levels of

389
P OST H AR V EST RI P ENING P H Y SIOLOG Y

ethylene in the storage environment and within the fruits as well (Xu, 1999;
Zambre et al., 2010; Rodoni et al., 2010). Exposure of O3 also reduced the
levels of pesticide in stored apples (Ong et al., 1996). Besides the practical
applicability of the above gases, required levels of O2, CO2, ethylene, humid-
ity, and condensation in the storage environment can also delay ripening
and improve the quality and storability of fruits (Mangaraj and Goswami,
2009; Hoehn et al., 2009; Yahia, 2009).
There are different endogenous gases and volatiles that play a
regulatory role in determining the production and response of ethylene
(Figure  12.3). Besides this, interactions of some of the volatiles, such as
C2H4, O2, CO2, SA/MeSA, JAs, and NO, on various components of the respi-
ratory metabolism were also reported (Ederli et  al., 2006; Leakey et  al.,
2009; Wang et al., 2010). This shows that different gases and volatiles can
affect the status and response of ethylene, along with a definite influence
on respiration, and this in turn is closely linked with the potential shelf life
of fruits after their harvest (Kader and Saltveit, 2003b; Paul and Srivastava,
2006; Paul et al., 2010b; Varoquaux and Ozdemir, 2005).

12.9  Conclusions and Future Perspectives

Endogenous volatiles in fruits are usually known for their role in determin-
ing the flavor and aroma. But in a recent trend, their importance and role
in fruit ripening and storability have come up. Combined and interactive
effects of different endogenous volatiles have now been studied with respect
to the postharvest physiology of fruits, ripening process, and postharvest
management practices. Now it is clear that both the external and internal
gaseous environments of the fruit play a crucial role in regulating the pro-
cess of ripening. Ethylene is, of course, a major controlling factor for the
ripening and related changes in climacteric fruits, but its biosynthesis, per-
ception, sensitivity, and even action are influenced either directly or indi-
rectly by other endogenous volatiles, such as acetaldehyde, ethanol, methyl
salicylate, methyl jasmonate, and NO, besides the O2 -to-CO2 ratio and water
status of the fruit. It is also quite apparent and possible that some volatiles
directly, indirectly, or in an interactive way may affect development and act
as developmental signals. Besides this, endogenous volatiles can also be
pivotal in altering the sensitivity of tissues toward ethylene and other plant
hormones, which is usually seen with the development of fruit. So far, not
much is understood about the interactive mechanisms of different volatiles
in regulating the process of ripening. The internal atmospheres in fruit
should therefore be analyzed and understood to enable effective manipu-
lation of ripening and control of the quality of fruits. Efforts to decipher
the dynamics of internal atmospheres in relation to the progress of fruit

390
 INTERNAL ATMOS P H ERE O F F RUITS

ripening could prove to be of great practical importance. Such information

would enable us to manage the storage environment and also optimize the
composition of the internal atmosphere in the fruit more precisely. This will
not only help in achieving the best quality, aroma, flavor, and shelf life, but
also assist in minimizing the decay, pest infestation, and microbial infection
of fruits during storage.
In our opinion, future studies on storage technology need to focus
on defining and then generating optimum internal atmospheres in fruits,
providing conditions that are cultivar specific, making use of precise sen-
sors for monitoring the levels of volatiles and gases, developing and using
a fruit-specific and real-time system to manipulate the levels of at least
some of the most critical volatiles and gases during storage, and mak-
ing use of some of the fruit-specific volatiles as exogenous treatments to
tackle both a specific or a broad problem related to postharvest storage
and management.

References
Abeles, F.B., and Gahagan, H.E. 1968. Abscission: the role of ethylene,
ethylene analogues, carbon dioxide, and oxygen. Plant Physiology 43,
1255–1258.
Abeles, F.B., and Takeda, F. 1990. Cellulase activity and ethylene in ripening
strawberry and apple fruits. Scientia Horticulturae 42, 269–275.
Abeles, F.B., Morgan, P.W., and Saltveit, M.E. 1992. Ethylene in Plant Biology.
2nd ed., vol. 15. Academic Press, San Diego.
Adato, I., and Gazit, S. 1974. Water-deficit stress, ethylene production and
ripening in avocado fruits. Plant Physiology 53, 45–46.
Alexander, L., and Grierson, D. 2002. Ethylene biosynthesis and action in
tomato: A model for climacteric fruit ripening. Journal of Experimental
Botany 53, 2039–2055.
Alonso, J.M., Chamarro, J., and Granell, A. 1995. Evidence for the involve-
ment of ethylene in the expression of specific RNAs during matura-
tion of the orange, a nonclimacteric fruit. Plant Molecular Biology 29,
385–390.
Amarante, C., Banks, N.H., and Ganesh, S. 2001. Characterizing ripening
behaviour of coated pears in relation to fruit internal atmosphere.
Postharvest Biology and Technology 23, 51–59.
Arroyo, F.T., Moreno, J., Daza, P., Boianova, L., and Romero, F. 2007. Antifungal
activity of strawberry fruit volatile compounds against Colletotrichum
acutatum. Journal of Agricultural and Food Chemistry 55, 5701–5707.
Asoda, T., Terai, H., Kato, M., and Suzuki, Y. 2009. Effects of postharvest etha-
nol vapor treatment on ethylene responsiveness in broccoli. Postharvest
Biology and Technology 2, 216–220.

391
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Aubert, C., and Chanforan, C. 2007. Postharvest changes in physicochemi-

cal properties and volatile constituents of apricot (Prunus armeniaca L.)
characterization of 28 cultivars. Journal of Agricutural and Food
Chemistry 55, 3074–3082.
Ayala-Zavala, J.F., Wang, S.Y., Wang, C.Y., and Gonzalez-Aguilar, G.A. 2005.
Methyl jasmonate in conjunction with ethanol treatment increases anti-
oxidant capacity, volatile compounds and postharvest life of strawberry
fruit. European Food Research and Technology 221, 731–738.
Bai, J., Hagenmaier, R.D., and Baldwin, E.A. 2003. Coating selection for
‘Delicious’ and other apples. Postharvest Biology and Technology 28,
381–390.
Baldwin, E.A. 2004. Flavor. In The Commercial Storage of Fruits, Vegetables
and Florist and Nursery Stocks, Gross, K.C., Wang, C.Y., and Saltveit,
M.E. (eds.). Agriculture Handbook Number 66. Agricultural Research
Service, Beltsville, MD, 18. http://usna.usda.gov/hb66/023flavor.pdf.
Baldwin, E.A., Burn, J.K., Kazokas, W., Brecht, J.K., Hagenmaier, R.D.,
Bender, R.J., and Pesis, E. 1999. Effect of two edible coatings with dif-
ferent permeability characteristics on mango (Mangifera indica L.) rip-
ening during storage. Postharvest Biology and Technology 17, 215–226.
Baldwin, E.A., Nisperos-Carriedo, M.O., and Moshonas, M.G. 1991.
Quantitative analysis of flavour and other volatiles and for certain con-
stituents of two tomato cultivars during ripening. Journal of the American
Society for Horticultural Science 116, 265–269.
Baldwin, E.A., Scott, J.W., Shewmaker, C.K., and Schuch, W. 2000. Flavour
trivia and tomato aroma: biochemistry and possible mechanisms for
control of important aroma components. HortScience 35, 1013–1022.
Banks, N.H. 1984. Internal atmosphere modification in Pro-Long coated
apples. Acta Horticulturae 157, 105–112.
Banks, N.H., and Nicholson, S.E. 2000. Internal atmosphere composition
and skin permeance to gases of pepper fruit. Postharvest Biology and
Technology 18, 33–41.
Bargel, H., and Neinhuis, C. 2005. Tomato (Lycopessicon esculentum Mill.)
fruit growth and ripening as related to the biomechanical properties
of fruit skin and isolated cuticle. Journal of Experimental Botany 56,
1049–1060.
Barry, C.S., and Giovannoni, J.J. 2007. Ethylene and fruit ripening. Journal of
Plant Growth Regulation 26, 143–159.
Beaulieu, J.C., Peiser, G., and Saltveit, M.E. 1997. Acetaldehyde is a causal
agent responsible for ethanol induced ripening inhibition in tomato
fruits. Plant Physiology 113, 431–439.
Belding, R.D., Blankenship, S.M., Young, E., and Leidy, R.B. 1998. Composition
and variability of epicuticular waxes in apple cultivars. Journal of the
American Society for Horticultural Science 123, 348–356.
Ben-Yehoshua, S. 1987. Transpiration, water stress and gas exchange. In
Postharvest Physiology of Vegetables, Weichman, J. (ed.). Marcel Dekker,
New York, 113.

392
 INTERNAL ATMOS P H ERE O F F RUITS

Ben-Yehoshua, S., and Rodov, V. 2003. Transpiration and water stress. In

Postharvest Physiology and Pathology of Vegetable, Bartz, J.A., and Brecht,
J.K. (eds.). Marcel Dekker, New York, 111–159.
Ben-Yehoshua, S., Burg, S.P., and Young, R. 1985. Resistance of citrus fruit
to mass transport of water vapours and gases. Plant Physiology 79,
1048–1053.
Ben-Yehoshua, S., Shapiro, B., Even-Chen, Z., and Lurie, S. 1983. Mode of
action of plastic film extending life of lemon and bell pepper fruit by
alleviation of water stress. Plant Physiology 73, 87–93.
Biale, J.B. 1964. Growth, maturation and senescence in fruits. Science 146,
880–888.
Biale, J.B., and Young, R.E. 1981. Respiration and ripening in fruits—­
Retrospective and prospect. In Recent Advances in the Biochemistry of
Fruits and Vegetables, Friend, J., and Rhodes, M.J.C. (eds.). Academic
Press, New York, 1–39.
Biles, C.L., Wall, M.M., and Blackstone, K. 1993. Morphological and physi-
ological changes during maturation of New Mexican type peppers.
Journal of the American Society for Horticultural Science 118, 476–480.
Blanke, M.M. 1986. Comparative SEM study of stomata on developing
quince, apple, grape and tomato fruit. Angewardte Botanik 60, 209–214.
Bleeker, P.M., Diergaarde, P.J., Ament, K., Guerra, J., Weidner, M., Schutz,
S., de Both, M.T.J., Haring, M.A., and Schuurink, R.C. 2009. The role of
specific tomato volatiles in tomato–whitefly interaction. Plant Physiology
151, 925–935.
Boersig, M.R., Kader, A.A., and Romani, R.J. 1988. Aerobic-anaerobic respira-
tory transition in pear fruit and cultured pear fruit cells. Journal of the
American Society for Horticultural Science 111, 869–873.
Borisjuk, L., Macherel, D., Benamar, A., Wobus, U., and Rolletschek, M.
2007. Low oxygen sensing and balancing in plant seeds—A role for
nitric oxide. New Phytology 176, 813–823.
Bouwmeester, H.J., Gershenzon, J., Konings, M.C.J.M., and Croteau, R. 1998.
Biosynthesis of the monoterpenes limonene and carvone in the fruit of
caraway. I. Demonstration of enzyme activities and their changes with
development. Plant Physiology 117, 901–912.
Bouzayen, M., Latche, A., Pech, J.C., and Nath, P. 2010. Mechanism of fruit
ripening. In Plant Developmental Biology—Biotechnological Perspective,
Pua, E.C., and Davey, M.R. (eds.). Vol. 1. Springer-Verlag, Heidelberg,
319–339.
Bower, J., Holford, P., Latche, A., and Pech, J.C. 2002. Culture conditions and
detachment of the fruit influence the effect of ethylene on the climac-
teric respiration of the melon. Postharvest Biology and Technology 26,
135–146.
Brauss, M.S., Linforth, R.S.T., and Taylor, A.J. 1998. Effect of variety, time
of eating and fruit to fruit variation on volatile release during eating of
tomato fruit (Lycopersicon esculentum). Journal of Agricultural and Food
Chemistry 46, 2287–2292.

393
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Bufler, G. 1984. Ethylene enhanced 1-aminocyclopropane-1-carboxylic acid

synthase activity in ripening apples. Plant Physiology 75, 192–196.
Bufler, G., and Bangerth, F. 1982. Pyruvate decarboxylase in “Golden
Delicious” apples: Kinetics and relation to acetone and ethanol pro-
duction in different storage atmospheres. Scientia Horticulturae 16,
137–146.
Burg, S.P., and Burg, E.A. 1962. The role of ethylene in fruit ripening. Plant
Physiology 37, 179–189.
Burg, S.P., and Burg, E.A. 1965a. Ethylene action and the ripening of fruits.
Science 148, 1190–1196.
Burg, S.P., and Burg, E.A. 1965b. Gas exchange in fruits. Physiologia
Plantarum 18, 870–886.
Burg, S.P., and Burg, E.A. 1967. Molecular requirement for the biological
activity of ethylene. Plant Physiology 42, 144–152.
Buttery, R. 1993. Quantitative and sensory aspects of flavour of tomato and
other vegetables and fruits. In Flavour Science: Sensible Principles and
Techniques, Acree, T.E., and Teranishi, R. (eds.). American Chemical
Society, Washington, DC, 259–286.
Cadwallader, K.R. 2005. Flavour and volatile metabolism in produce. In
Produce Degradation Pathways and Prevention, Lamikanra, O., Imam,
S., and Ukaku, D. (eds.). CRC Press, Taylor & Francis Group, Boca
Raton, FL, 155–180.
Calbo, A.G., Pererira, A.S., and Horino, Y. 1988. Sealants for ripening con-
trol of tomatoes. In Brazilian Congress of Vegetable Crops: Proceedings of
SOB, Brazil, vol. 6, p. 28 [in Portuguese].
Cameron, A.C., and Yang, S.F. 1982. A simple method for the determina-
tion of resistance to gas diffusion in plant organs. Plant Physiology 70,
21–23.
Cancel, J.D., and Larsen, P.B. 2002. Loss-of-function mutations in the ethylene
receptor ETR1 cause enhanced sensitivity and exaggerated response to
ethylene in Arabidopsis. Plant Physiology 129, 1557–1567.
Chaves, A.R., and Tomas, J.O. 1984. Effect of a brief CO2 exposure on ethyl-
ene production. Plant Physiology 76, 88–91.
Chavez-Franco, S.H., and Kader, A.A. 1993. Effects of CO2 on ethylene
biosynthesis in ‘Bartlett’ pears. Postharvest Biology and Technology 3,
183–190.
Chen, A.R.S., and Chase, J., Jr. 1993. Alcohol dehydrogenase and pyruvate
decarboxylase induction in ripening and hypoxia tomato fruit. Plant
Physiology and Biochemistry 31, 875–885.
Chervin, C., El-Kereamy, A., Roustan, J.P., Latche, A., Lamon, J., and
Bouzayen, M. 2004. Ethylene seems required for the berry develop-
ment and ripening in grape, a non-climacteric fruit. Plant Science 167,
1301–1305.
Chervin, C., Lavigne, D.L., and Westercamp, P. 2009. Reduction in gray mold
development in table grapes by preharvest sprays with ethanol and cal-
cium chloride. Postharvest Biology and Technology 54, 115–117.

394
 INTERNAL ATMOS P H ERE O F F RUITS

Chervin, C., Tira-Umphon, A., Terrier, N., Zouine, M., Severac, D., and
Roustan, J.P. 2008. Stimulation of the grape berry expansion by ethyl-
ene and effects on related gene transcripts, over the ripening phase.
Physiologia Plantarum 334, 534–546.
Crisosto, C.H., Garner, D., and Crisosto, G. 2002. Carbon dioxide-enriched
atmosphere during cold storage limit losses from Botrytis but acceler-
ate rachis browning of ‘Redglobs’ table grapes. Postharvest Biology and
Technology 26, 181–189.
Dadzie, B.K., Banks, N.H., Cleland, D.S., and Hewett, E.W. 1993. Role of skin
resistance to gas diffusion in the response of fruits to modified atmo-
sphere. Acta Horticulturae 343, 129–134.
Dandekar, A.M., Teo, G., Defilippi, B.G., Uratsu, S.L., Passey, A.J., Kader,
A.A., Stow, J.R., Colgan, R.J., and James, D.J. 2004. Effect of down-­
regulation of ethylene biosynthesis on fruit flavor complex in apple
fruit. Transgenic Research 13, 373–384.
Defilippi, B.G., Manriquez, D., Luengwilai, K., and Gonzalez-Aguero, M.
2009. Aroma volatiles: biosynthesis and mechanisms of modulation dur-
ing fruit ripening. Advances in Botanical Research 50, 1–37.
de Leon-Sanchez, F.D., Pelayo-Zaldivar, C., Rivera-Cabrera, F., Ponce-
Valadez, M., Avila-Alejandre, X., Fernandez, F.J., Escalona-Buendia,
H.B., and Perez-Flores, L.J. 2009. Effect of refrigerated storage on
aroma and alcohol dehydrogenase activity in tomato fruit. Postharvest
Biology and Technology 54, 93–100.
Devoto, A., and Turner, J.G. 2005. Jasmonate—Regulated Arabidopsis stress
signalling network. Physiologia Plantarum 123, 161–172.
de Vries, H.S.M., Harren, F.J.M., Voesenek, L.A.C.J., Blom, C.W.P.M.,
Woltering, E.J., van der Valk, H.C.P.M., and Reuss, J. 1995. Investigation
of local ethylene emission from intact cherry tomatoes by means of pho-
tothermal deflection and photoacoustic detection. Plant Physiology 107,
1371–1377.
Dietz, T.H., Timma-Raju, K.R., and Joshi, S.S. 1988. Studies on loss of weight
of mango fruits as influenced by cuticle and lenticels. Acta Horticulturae
231, 457–460.
Ding, C.K., and Wang, C.Y. 2003. The dual effects of methyl salicylate on
ripening and expression of ethylene biosynthetic genes in tomato fruit.
Plant Science 164, 589–596.
Ding, C.K., Wang, C., Gross, K., and Smith, D. 2002. Jasmonate and salic-
ylate induce the expression of pathogenesis-related-protein genes
and increase resistance to chilling injury in tomato fruit. Planta 214,
895–901.
Dudareva, N., and Pichersky, E. 2000. Biochemical and molecular genetic
aspects of floral scents. Plant Physiology 122, 627–633.
Dudareva, N., Pichersky, E., and Gershenzon, J. 2004. Biochemistry of plant
volatiles. Plant Physiology 135, 1893–1902.
Eaks, I.L. 1970. Respiratory response, ethylene production, and response to
ethylene of citrus fruit during ontogeny. Plant Physiology 45, 334–338.

395
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Ederli, L., Morettini, R., Borgogni, A., Wasternack, C., Miersch, O., Reale, L.,
Ferranti, F., Tosti, N., and Pasqualini, S. 2006. Interaction between nitric
oxide and ethylene in the induction of alternative oxidase in ozone-
treated tobacco plants. Plant Physiology 142, 595–608.
Ehret, D.L., Helmer, T., and Hall, J.W. 1993. Cuticle cracking in tomato fruit.
Journal of Horticultural Science 68, 195–201.
El-Kereamy, A., Chervin, C., Roustan, J.P., Cheynier, V., Souquet, J.M.,
Moutounet, M., Raynal, J., Ford, C., Latche, A., Pech, J.C., and Bouzayen,
M. 2003. Exogenous ethylene stimulates the long-term expression of
genes related to anthocyanin biosynthesis in grape berries. Physiologia
Plantarum 119, 175–182.
Elyatem, S.M., Banks, N.H., and Cameron, A.C. 1994. Oxygen concentration
effects on ethylene production by ripening banana tissue. Postharvest
Biology and Technology 4, 343–351.
Eum, H.L., Kim, H.B., Choi, S.B., and Lee, S.K. 2009. Regulation of ethylene
biosynthesis by nitric oxide in tomato (Solanum lycopersicum L.) fruit
harvested at different ripening stages. European Food Research and
Technology 228, 331–338.
Fan, X., Mattheis, J.P., and Fellma, J.K. 1998. A role for jasmonates in climac-
teric fruit ripening. Planta 204, 444–449.
Finger, F.L., Puschmann, R., and Barros, R.S. 1995. Effects of water loss on
respiration: Ethylene production and ripening of banana fruit. Revista
Brasileira de Fisiologia Vegetal 7, 115–118.
Flores, F., El-Yahyaoui, F., de Billerbeck, G., Romojaro, F., Latche, A.,
Bouzayen, M., Pech, J.C., and Ambid, C. 2002. Role of ethylene in the
biosynthesis pathway of aliphatic ester aroma volatiles in Charentais
cantaloupe melons. Journal of Experimental Botany 53, 201–206.
Forney, C.F., Mathheis, J.P., and Austin, R.K. 1991. Volatile compounds pro-
duced by broccoli under anaerobic conditions. Journal of Agricultural
and Food Chemistry 39, 2257–2259.
Franck, C., Lammertyn, J., Ho, Q.T., Verboven, P., Verlinden, B., and Nicolai,
B.A. 2007. Browning disorders in pear: A review. Postharvest Biology
and Technology 43, 1–13.
Frenkel, C., Erez, A., and Henninger, M.R. 1995. Ethanol induced cold toler-
ance in chilling-sensitive crops. In Harvest and Post Harvest Technologies
for Fresh Fruits and Vegetables: Proceedings of the International
Conference, Guanajuanta, Mexico, February 20–24, pp. 512–521.
Gamage, T.V., and Rahman, M.S. 1999. Postharvest handling of foods of plant
origin. In Handbook of Food Preservation, Rahman, M.S. (ed.). Marcel
Dekker, New York, 11–46.
Gelly, M., Recasens, I., Mata, M., Arbones, A., Rufat, J., Girona, J., and Marsal,
J. 2003. Effect of water deficit during stage II of peach fruit development
and post harvest on fruit quality and ethylene production. Journal of
Horticultural Science and Biotechnology 78, 324–330.
Genard, M., and Gouble, B. 2005. ETHY. A theory of fruit climacteric ethyl-
ene emission. Plant Physiology 139, 531–545.

396
 INTERNAL ATMOS P H ERE O F F RUITS

George, J.B., Marriott, J., Palmer, J.M., and Karikary, S.K. 1982. Sensitivity
to water stress and ethylene of stored plantain fruits. Journal of
Experimental Botany 33, 1194–1201.
Gershenzon, J., McConkey, M.E., and Croteau, R.B. 2000. Regulation of
monoterpene accumulation in leaves of peppermint. Plant Physiology
122, 205–213.
Giovannoni, J. 2001. Molecular biology of fruit maturation and ripening.
Annual Review of Plant Physiology and Plant Molecular Biology 52,
725–749.
Golding, J.B., Shearer, D., McGlasson, W.B., and Wyllie, S.G. 1999.
Relationship between respiration, ethylene and aroma production
in ­ripening banana. Journal of Agricultural and Food Chemistry 47,
1646–1651.
Goldschmidt, E.E. 1997. Ripening of citrus and other non-climacteric fruits: A
role for ethylene. Acta Horticulturae 463, 335–340.
Gorny, J.R., and Kader, A.K. 1996. Controlled atmosphere suppression of
ACC-synthase and ACC-oxidase in ‘Golden Delicious’ apple during
long-term cold storage. Journal of the American Society for Horticultural
Science 121, 751–755.
Gran, C.D., and Beaudry, R.M. 1993. Determination of the low oxygen limit
for several commercial apple cultivars by respiratory quotient break
point. Postharvest Biology and Technology 3, 259–267.
Grierson, D., and Wardowski, W.F. 1978. Relative humidity effects on the post
harvest life of fruits and vegetable. HortScience 13, 570–574.
Gross, K.C., Watada, A.E., Kang, M.S., Kim, S.D., and Lee, S.W. 1986.
Biochemical changes associated with the ripening of hot pepper fruit.
Physiologia Plantarum 66, 31–36.
Guichad, E., and Souty, M. 1988. Comparison of the relative quantities of
aroma compounds found in fresh apricot (Prunus armeniaca) from
six different varieties. Zeitschrift für Lebensmittel-Untersuchung und
-Forschung 186, 301–307.
Gustafson, F.G. 1934. Production of alcohol and acetaldehyde by tomatoes.
Plant Physiology 9, 359–367.
Hadfield, K.A., Rose, J.K.C., and Bennett, A.B. 1995. The respiratory climac-
teric is present in Charentais (Cucumis melo cv. Reticulatus F1 Alpha)
melons ripened on or off the plant. Journal of Experimental Botany 46,
1923–1925.
Hagenmaier, R. 2004. Method for measuring internal gases of citrus fruit
and determining peel permeance. Proceedings of the Florida State
Horticulture Society 116, 418–423.
Hagenmaier, R.D. 2005. A comparison of ethane, ethylene and CO2 peel
permeance for fruits with different coatings. Postharvest Biology and
Technology 37, 56–64.
Hansen, M., Buttery, R.G., Stern, D.J., Cantwell, M.I., and Ling, L.C. 1992.
Broccoli storage under low oxygen atmosphere: Identification of higher
boiling volatiles. Journal of Agricultural and Food Chemistry 40, 850–852.

397
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Harpster, M.H., Lee, K.Y., and Dunsmuir, P. 1997. Isolation and character-
ization of gene encoding endo-β-1,4-glucanase from pepper (Capsicum
annuum L.). Plant Molecular Biology 33, 47–59.
Hayata, Y., Maneerat, C., Kozuka, H., Sakamoto, K., and Ozajima, Y. 2002.
Flavor volatile analysis of ‘House Momotaro’ tomato fruit extract at dif-
ferent ripening stages by porapak Q column. Journal of the Japanese
Society for Horticultural Science 71, 473–479.
Ho, Q.T., Verboven, P., Mebatsion, H.K., Verlinden, B.E., Vandewalle, S., and
Nicolai, B.M. 2009. Microscale mechanisms of gas exchange in fruit tis-
sue. New Phytology 182, 162–174.
Ho, Q.T., Verboven, P., Verlinden, B.E., Lammertyn, J., Vandewalle, S., and
Nicolai, B.M. 2008. A continuum model for metabolic gas exchange in
pear fruit. PLoS Computational Biology 4, e1000023.
Ho, Q.T., Verboven, P., Verlinden, B.E., and Nicolai, B.M. 2010. A model for
gas transport in pear fruit at multiple scales. Journal of Experimental
Botany 61, 2071–2081.
Ho, Q.T., Verlinden, B.E., Verboven, P., and Nicolai, B.M. 2006a. Gas diffu-
sion properties at different positions in the pear. Postharvest Biology and
Technology 41, 113–120.
Ho, Q.T., Verlinden, B.E., Verboven, P., Vandewalle, S., and Nicolai, B.M.
2006b. A permeation-diffusion-reaction model of gas transport in cellular
tissue of plant material. Journal of Experimental Botany 57, 4215–4224.
Hoeberichts, F.A., Van Der Plas, L.H.W., and Woltering, E.J. 2002. Ethylene
perception is required for the expression of tomato ripening related
genes and associated physiological changes even at advanced stages of
ripening. Postharvest Biology and Technology 26, 125–133.
Hoehn, E., Prange, R.K., and Vigneault, C. 2009. Storage technology and
applications. In Modified and Controlled Atmospheres for the Storage,
Transportation, and Packaging of Horticultural Commodities, Yahia,
E.M. (ed.). CRC Press, Taylor & Francis Group, Boca Raton, FL, 17–50.
Huelin, F.E., and Coggiola, I.M. 1968. Superficial scald, a functional disorder
of stored apples. IV. Effect of variety, maturity, oiled wraps and diphe-
nylamine on the concentration of α-farnesene in the fruit. Journal of the
Science of Food and Agriculture 19, 297–301.
Hui, Y.H. (ed.). 2010. Handbook of Fruit and Vegetable Flavors. John Wiley &
Sons, Hoboken, NJ.
Hurr, B.M., Huber, D.J., Vallejos, C.E., and Talcott, S.T. 2009.
Developmentally dependent responses of detached cucumber
(Cucumis sativus L.) fruit to exogenous ethylene. Postharvest Biology
and Technology 52, 207–215.
Iannetta, P.P., Loorhoven, L.J., Davies, H.V., and Harren, F. 2000. Ethylene pro-
duction by strawberry flowers and the ripening fruit. In Communication
to the 9th International Workshop on LASER Based Photoacoustic Trace
Gas Detection in Life Science, Nijmegen, the Netherlands 11–12 July 2000.
Iannetta, P.P.M., Laarhoven, L.J., Medina-Escobar, N., James, E.K., McManus,
M.T., Davies, H.V., and Harren, F.J.M. 2006. Ethylene and carbon dioxide

398
 INTERNAL ATMOS P H ERE O F F RUITS

production by developing strawberries show a correlative pattern that

is indicative of ripening climacteric fruit. Physiologia Plantarum 127,
247–259.
Imanishi, S., Noguchi, A., Hatakeyama, R., and Nagata, M. 2005. Monitoring
the effect of jasmonates on the expression of ripening related genes
in tomato fruit disks by cDNA macroarray. Acta Horticulturae 682,
149–153.
Inaba, A. 2007. Studies on the internal feedback regulation of ethylene biosyn-
thesis and signal transduction during fruit ripening, and the improve-
ment of fruit quality. Journal of the Japanese Society for Horticultural
Science 76, 1–12.
Jackson, M.B., Herman, B., and Goodenough, A. 1982. An examination of the
importance of ethanol in causing injury to flooded plants. Plant Cell and
Environment 5, 163–172.
Jacob-Wilk, D., Holland, D., Goldschmidt, E.E., Riov, J., and Eyal, Y. 1999.
Chlorophyll breakdown by chlorophyllase: Isolation and functional
expression of the Chlase1 gene from ethylene treated citrus fruit and its
regulation during development. Plant Journal 20, 653–661.
Johnston, J.W., Gunaseelan, K., Pidakala, P., Wang, M., and Schaffer, R.J. 2009.
Co-ordination of early and late ripening events in apples is regulated
through differential sensitivities to ethylene. Journal of Experimental
Botany 60, 2689–2699.
Joyce, D.C., Hockings, P.D., Mazucco, R.A., and Shorter, A.J. 2002. H-1-
nuclear magnetic resonance imaging of ripening ‘Kensington Pride’
mango fruit. Functional Plant Biology 29, 873–879.
Kader, A.A. 1986. Biochemical and physiological basis for effects of controlled
and modified atmospheres on fruits and vegetables. Food Technology
40, 99–104.
Kader, A.A. 1987. Respiration and gas exchange of vegetable. In Postharvest
Physiology of Vegetable: Food Science and Technology, Weichmann, J.
(ed.). Marcel Dekker, New York, 25–43.
Kader, A.A. 2009. Future research and application needs. In Modified
and Controlled Atmospheres for Storage, Transpiration, Packaging of
Horticultural Commodities, Yahia, E.M. (ed.). CRC Press, Taylor &
Francis Group, Boca Raton, FL, 569–575.
Kader, A.A., and Barrett, D.M. 1996. Classification, composition of fruits
and postharvest maintenance of quality. In Processing Fruits: Science
and Technology, Biology, Principles and Applications, Somogyi, L.P.,
Ramaswamy, H.S., and Hui, Y.H. (eds.). Vol. 1. Technomic Publishing
Company, Lancaster, PA, 1–24.
Kader, A.A., and Morris, L.L. 1977. Relative tolerance of fruits and vegeta-
bles to elevated CO2 and reduced O2 levels. Michigan State University
Horticulture Report 28, 260–265.
Kader, A.A., and Saltveit, M.E. 2003a. Respiration and gas exchange. In
Postharvest Physiology and Pathology of Vegetables, Bartz, J.A., and
Brecht, J.K. (eds.). Marcel Dekker, New York, 7–29.

399
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Kader, A.A., and Saltveit, M.E. 2003b. Atmosphere modification. In Postharvest

Physiology and Pathology of Vegetables, Bartz, J.A., and Brecht, J.K. (eds.).
New York, Marcel Dekker, 229–246.
Kader, A.A., Zagory, D., and Kerbel, E.L. 1989. Modified atmosphere pack-
aging of fruits and vegetables. Critical Reviews in Food Science and
Nutrition 28, 1–30.
Kanellis, A.K., Loulakakis, K.A., Hassan, M., and Roubelakis-Angelakis, K.A.
1993. Biochemical and molecular aspects of the low oxygen action on
fruit ripening. In Cellular and Molecular Aspects of Biosynthesis and
Action of the Plant Hormone Ethylene, Pech, J.C., Latche, A., and Balaque,
C. (eds.). Kluwer Academic, Dordrecht, the Netherlands, 117–122.
Kanellis, A.K., Solomos, T., and Mattoo, A.K. 1989a. Hydrolytic enzyme activi-
ties and protein pattern of avocado fruit ripened in air and in low oxygen
with and without ethylene. Plant Physiology 90, 259–266.
Kanellis, A.K., Solomos, T., and Mattoo, A.K. 1989b. Changes in sugar, enzy-
matic activities and acid phophatase isozyme profile of bananas ripened
in air or stored in 2.5% O2 with and without ethylene. Plant Physiology
90, 251–258.
Kanellis, A.K., Solomos, T., and Roubelakis-Angelakis, K.A. 1991. Suppression
of cellulase and polygalacturonase and induction of alcohol dehydroge-
nase isoenzymes of avocado fruit mesocarp subjected to low oxygen
stress. Plant Physiology 96, 269–274.
Kanellis, A.K., Tonutti, P., and Perata, P. 2009. Biochemical and molecular
aspects of modified and controlled atmospheres. In Modified and
Controlled Atmospheres for Storage, Transpiration and Packaging of
Horticultural Commodities, Yahia, E.M. (ed.). CRC Press, Taylor &
Francis Group, Boca Raton, FL, 553–567.
Karl, T., Guenther, A., Turnipseed, A., Patton, E.G., and Jardine, K. 2008.
Chemical sensing of plant stress at the ecosystem scale. Biogeoscience
5, 1287–1294.
Katz, E., Lagunes, P.M., Riov, J., Weiss, D., and Goldschmidt, E.E. 2004.
Molecular and physiological evidence suggests the existence of a sys-
tem II-like pathway of ethylene production in non-climacteric citrus
fruit. Planta 219, 243–252.
Ke, D., and Kader, A.A. 1990. Tolerance of Valencia oranges to controlled atmo-
spheres as determined by physiological responses and quality attributes.
Journal of the American Society for Horticultural Science 115, 779–783.
Ke, D.L., Goldstein, L., O’Mohony, M., and Kader, A.A. 1991. Effect of short-
term exposure to low O2 and high CO2 atmospheres on quality attri-
butes of strawberries. Journal of Food Science 56, 50–54.
Kelly, M.O., and Saltveit, M.E. 1988. Effect of endogenously synthesized and
exogenously applied ethanol on tomato fruit ripening. Plant Physiology
88, 143–147.
Kendall, S.A., and Ng, T.J. 1988. Genetic variation of ethylene production in
harvested muskmelon fruit. HortScience 23, 759–761.

400
 INTERNAL ATMOS P H ERE O F F RUITS

Kimmerer, T.W., and Kozlowski, T.T. 1982. Ethylene, ethane, acetaldehyde,

and ethanol production by plants under stress. Plant Physiology 69,
840–847.
Klee, H.J. 2002. Control of ethylene mediated processes in tomato at the level
of receptors. Journal of Experimental Botany 53, 2057–2063.
Klieber, A., Ratanachinakorn, B., and Simone, D.H. 1996. Effect of low oxy-
gen and high carbon dioxide on tomato cultivar ‘Bermuda’ fruit physiol-
ogy and composition. Scientia Horticulturae 65, 251–261.
Klozenbucher, K.A., Altman, S.A., McIntosh, M.S., and Walsh, C.S. 1994.
Effect of cultivar on endogenous ethylene evolution and its relationship
to increase of soluble protein in peach mesocarp tissue. Fruit Variety
Journal 48, 20–26.
Knee, M. 1982. Fruit softening III. Requirement for oxygen and pH effects.
Journal of Experimental Botany 33, 1263–1269.
Kondo, S., Tomiyama, A., and Seto, H. 2000. Changes of endogenous jas-
monic acid and methyl jasmonate in apples and sweet cherries during
fruit development. Journal of the American Society for Horticultural
Science 125, 282–287.
Kovacs, E., Kovacs, J., Zackel, E., and Genova, J. 1994. Structural and chemi-
cal changes of pear skin. Acta Horticulturae 368, 243–250.
Kubo, Y., Inaba, A., and Nakamura, R. 1996. Extinction point and critical
oxygen concentration in various fruits and vegetables. Journal of the
Japanese Society for Horticultural Science 65, 397–402.
Lammertyn, J., Scheerlinck, N., Jancsok, P., Verlinden, B.E., and Nicolai, B.M.
2003. A respiration-diffusion model for ‘Conference’ pears II: stimula-
tion and relation to core breakdown. Postharvest Biology and Technology
30, 43–55.
Lanciotti, R., Belletti, N., Patrignani, F., Gianotti, A., Gardini, F., and Guerzoni,
M.E. 2003. Application of hexanal, (E)-2-hexenal, and hexyl acetate to
improve the safety of fresh-sliced apples. Journal of Agricultural and
Food Chemistry 51, 2958–2963.
Lange, D.L., and Kader, A.A. 1997. Effects of elevated carbon dioxide on key
mitochondrial respiratory enzymes in ‘Hass’ avocado fruit and fruit
disks. Journal of the American Society for Horticultural Science 122,
228–244.
Larsen, M. 1994. Volatile compounds formed in strawberries under anaerobic
conditions and their influence on self-flavour formation. In Trends in
Flavour Research, Maarse, H., and van der Heij, D.G. (eds.). Elsevier,
Amsterdam, 421–424.
Larsen, M., and Poll, L. 1990. Odour threshold of some important aroma com-
pounds in raspberries. Zeitschrift für Lebensmittel-Untersuchung und
-Forschung 191, 129–131.
Lawes, G.S., and Prasad, L. 1999. Peel permeance and storage changes in
internal atmosphere composition of surface-coated mandarin. Acta
Horticulture 485, 249–254.

401
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Leakey, A.D.B., Xu, F., Gillespie, K.M., McGrath, J.M., Ainsworth, E.A., and
Ort, D.R. 2009. Genomic basis for stimulated respiration by plants grow-
ing under elevated carbon dioxide. Proceedings of the National Academy
of Sciences USA 106, 3597–3602.
Lebrun, M., Plotto, A., Goodner, K., Ducamp, M.N., and Baldwin, E. 2008.
Discrimination of mango fruit maturity by volatiles using the electronic
nose and gas chromatography. Postharvest Biology and Technology 48,
122–131.
Lelievre, J.M., Latche, A., Jones, B., Bouzayen, M., and Peach, J.C. 1997.
Ethylene and fruit ripening. Physiologia Plantarum 101, 727–739.
Leonard, E.R., and Wardlaw, C.W. 1941. Studies in tropical fruit. XII. The res-
piration of banana during storage at 53F and ripening at controlled tem-
peratures. Annals of Botany 5, 379–423.
Leshem, Y.Y., Huang, J.S., Tzeng, D.S., and Chou, C.C. 2000. Nitric oxide gas
as an endogenous regulator of fruit, vegetable and flower maturation
and senescence. In Nitric Oxide in Plants: Occurrence, Function and Use,
Ledhem, Y.Y. (ed.). Kluwer Academic, Dordrecht, the Netherlands, 33–62.
Leshem, Y.Y., Wills, R.B.H., and Ku, V.V.V. 1998. Evidence for the function
of the radical gas nitric oxide (NO) as an endogenous maturation and
senescence regulating factor in higher plants. Plant Physiology and
Biochemistry 36, 825–833.
Leshuk, J.A., and Saltveit, M.E., Jr. 1990. Controlled atmosphere storage
requirement and recommendations for vegetables. In Food Preservation
by Modified Atmosphere, Calderon, M., and Barkai-Golan, R. (eds.). CRC
Press, Boca Raton, FL, 315–352.
Leslie, C.A., and Romani, R.J. 1986. Salicylic acid: A new inhibitor of ethylene
biosynthesis. Plant Cell Reports 5, 144–146.
Lewinsohn, E., Schalechet, F., Wilkinson, J., Matsui, K., Tadmor, Y., Nam,
K.H., Amar, O., Lastochkin, E., Larkov, O., Ravid, U., Hiatt, W., Gepstein,
S., and Pichersky, E. 2001. Enhanced levels of the aroma and flavor com-
pound S-linalool by metabolic engineering of the terpenoid pathway in
tomato fruits. Plant Physiology 127, 1256–1265.
Littmann, M.D. 1972. Effect of water stress on the ripening of climacteric
fruits. Queensland Journal of Agriculture and Animal Science 29,
103–113.
Longhurst, T.J., Lee, E., Hinde, R., Brady, C.J., and Speirs, J. 1994. Structure
of the tomato Adh2 gene and Adh2 pseudogenes and a study of Adh2
gene expression in fruit. Plant Molecular Biology 26, 1073–1084.
Lorenzo, O., Piqueras, R., Sanchez-Serrano, J.J., and Solano, R. 2003. Ethylene
response factor 1 integrates signal from ethylene and jasmonate path-
ways in plant defense. Plant Cell 15, 165–178.
Loreto, F., and Schnitzler, J.P. 2010. Abiotic stresses and induced BVOCs.
Trends in Plant Science 15, 154–166.
Lougheed, E.C., Murr, D.P., and Toivonen, P.M.A. 1987. Ethylene and non-
ethylene volatiles. In Postharvest Physiology of Vegetables, Weichmann, J.
(ed.). Marcel Dekker, New York, 255–276.

402
 INTERNAL ATMOS P H ERE O F F RUITS

Lurie, S., Pre-Aymard, C., Ravid, U., Larkov, O., and Fallik, E. 2002. Effect
of 1-methylcyclopropene on volatile emission and aroma in cv. Anna
apples. Journal of Agricultural and Food Chemistry 50, 4251–4256.
Lyons, J.M., and Pratt, H.K. 1964. Effect of stage of maturity and ethylene
treatment on respiration and ripening of tomato fruits. Journal of the
American Society for Horticultural Science 84, 491–500.
Mangaraj, S., and Goswami, T.K. 2009. Modified atmosphere packaging—
An ideal food preservation technique. Journal of Food Science and
Technology 46, 399–410.
Maquire, K.M., Banks, N.H., and Opara, L.U. 2001. Factors affecting weight
loss of apples. Horticultural Reviews 25, 197–233.
Mathooko, F.M. 1996. Regulation of ethylene biosynthesis in higher plants by
carbon dioxide. Postharvest Biology and Technology 7, 1–26.
Mathooko, F.M., Kubo, Y., Inaba, A., and Nakamura, R. 1995. Characterization
of the regulation of ethylene biosynthesis in tomato fruit by carbon
dioxide and diazocyclopentadiene. Postharvest Biology and Technology
5, 221–233.
Mathooko, F.M., Tsunasima, Y., Owino, W., Kubo, Y., and Inaba, A. 2001.
Regulation of genes encoding ethylene biosynthesis in peach fruit by
carbon dioxide and 1-methylcyclopropene. Postharvest Biology and
Technology 21, 265–281.
McDonald, R.E., McCollum, T.G., and Baldwin, E.A. 1996. Prestorage heat
treatments influence, free sterols and flavour volatiles of tomatoes stored
at chilling temperature. Journal of the American Society for Horticultural
Science 121, 531–536.
McMurchie, E.J., McGlasson, W.B., and Eaks, I.L. 1972. Treatment of fruit
with propylene gives information about biogenesis of ethylene. Nature
237, 235–236.
Mendoza, F., Verboven, P., Mebatsion, H.K., Kerckhofs, G., Wevers, M., and
Nicolai, B. 2007. Three-dimensional pore space quantification of apple
tissue using x-ray computed microtomography. Planta 226, 559–570.
Miccolis, V., and Saltveit, M.E., Jr. 1991. Morphological and physiologi-
cal changes during fruit growth and maturation of seven melon cul-
tivars. Journal of the American Society for Horticultural Science 116,
1025–1029.
Mishra, B., Khatkar, B.S., Garg, M.K., and Wilson, L.A. 2010. Permeability
of edible coating. Journal of Food Science and Technology 47, 109–113.
Mishra, V.K., and Gamage, T.V. 2007. Postharvest physiology of fruits and
vegetables. In Handbook of Food Preservation, Rahman, M.S. (ed.). 2nd
ed. CRC Press, Taylor & Francis Group, Boca Raton, FL, 19–48.
Mulas, M. 1994. Genetic variability of histological characteristics in olive
fruits. Acta Horticulturae 356, 70–73.
Musse, M., Guio, F.D., Quellec, S., Cambert, M., Challois, S., and Davenel,
A. 2010. Quantification of microporosity in fruit by MRI at various
magnetic fields: Comparison with x-ray microtomography. Magnetic
Resonance Imaging 28, 1525–1534.

403
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Nagamani, J.E., Shivashankara, K.S., and Roy, T.K. 2010. Role of oxidative
stress and activity of ethylene biosynthetic enzymes on formation
of spongy tissue in ‘Alphonso’ mango. Journal of Food Science and
Technology 47, 295–299.
Nagata, M., Mori, H., Tabei, Y., Sato, T., Hirai, M., and Imaseki, H. 1995.
Modification of tomato fruit ripening by transformation with sense or
antisense chimeric 1-aminocyclopropane-1-carboxylate synthase gene.
Acta Horticulturae 394, 213–218.
Nakano, R., Inoue, S., Kubo, Y., and Inaba, A. 2002. Water stress-induced eth-
ylene in the calyx trigger autocatalytic ethylene production and fruit
softening in ‘Tone-wase’ persimmon grown in a heated plastic-house.
Postharvest Biology and Technology 25, 293–300.
Nakatsuka, A., Murachi, S., Okunishi, H., Shiomi, S., Nakano, R., Kubo, Y., and
Inaba, A. 1998. Differential expression and internal feedback regulation
of l-aminocyclopropane-l-carboxylate synthase, 1-aminocyclopropane-­
l-carboxylate oxidase and ethylene receptor genes in tomato fruit dur-
ing development and ripening. Plant Physiology 118, 1295–1305.
Nath, A., Deka, B.C., Singh, A., Patel, R.K., Paul, D., Mishra, L.K., and Ojha,
H. 2011. Extension of shelf life of pear fruits using different packaging
materials. Journal of Food Science and Technology 49, 556–563.
Negre-Zakharov, F., Long, M.C., and Dudareva, N. 2009. Floral scents and
fruit aromas inspired by nature. In Plant-Derived Natural Products:
Synthesis, Function and Application, Osbourn, A.E., and Lanzotti, V.
(eds.). Springer, Dordrecht, 405–434.
Nguyen, T.A., Verboven, P., Daudin, J.D., Vandewalla, S., and Nicolai, B.M.
2004. Effect of picking date, time and temperature on water sorption
of ‘Conference’ pear tissue. Postharvest Biology and Technology 33,
243–253.
Niemirow-Krizsai, B., and Csillag, A. 1994. Anatomical reference to storability
in tomato berries. Acta Horticuturae 368, 310–315.
Nuevo, P.A., Lizada, M.C.C., and Pantastico, Er.B. 1984. Gas diffusion factors
in fruits III. Oxygen and carbon dioxide. Postharvest Research Notes 1,
86–89.
Obando, J., Miranda, C., Jowkar, M.M., Moreno, E., Souri, M.K., Martinez,
I.A., Arus, P., Garcia-Mas, J., Monforte, A.J., and Fernandez-Trujillo, J.P.
2007. Creating climacteric melon fruit from non-climacteric parentals:
postharvest quality implications. In Advances in Plant Ethylene Research:
Proceedings of the 7th International Symposium on Plant Hormone
Ethylene, Ramina, A., Chang, J., Giovannoni, J., Klee, H., Perata, P., and
Woltering, E. (eds.). Kluwer Academic, Dordrecht, the Netherlands,
197–205.
Obando-Ulloa, J.M., Jowkar, M.M., Moreno, E., Souri, M.K., Dos-Santos, N.,
Sanmarti, P., Bueso, M.C., Kessle, M., Martine, J.A., Alarcon, A., Nicolai,
B., Lammertyn, J., Garcia-Mas, J., Monforte, A.J., and Fernandez-Trujill,
J.P. 2008a. Near-isogenic lines of melon with different climacteric

404
 INTERNAL ATMOS P H ERE O F F RUITS

behavior as a tool to characterize fruit senescence trait. In Proceedings of

the IXth EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae,
Avignon, France, May 21–24, pp. 109–113.
Obando-Ulloa, J.M., Moreno, E., Garcia-Mas, J., Nicolai, B., Lammertyn,
J., Monforte, A.J., and Fernandez-Trujillo, J.P. 2008b. Climacteric or
non-climacteric behavior in melon fruit 1. Aroma volatiles. Postharvest
Biology and Technology 49, 27–37.
Obando-Ulloa, J.M., Nicolai, B., Lammertyn, J., Bueso, M.C., Monforte, A.J.,
and Fernandez-Trujillo, J.P. 2009. Aroma volatiles associated with the
senescence of climacteric or non-climacteric melon fruit. Postharvest
Biology and Technology 52, 146–155.
Ong, K.C., Cash, J.N., Zabik, M.J., Siddiq, M., and Jones, A.L. 1996. Chlorine
and ozone washes for pesticide removal from apples and processed
apple sauce. Food Chemistry 55, 153–160.
Pandit, S.S., Kulkarni, R.S., Chidley, H.G., Giri, A.P., Pujari, K.H., Kollner, T.G.,
Degenhardt, J., Gershenzon, J., and Gupta, V.S. 2009. Changes in vola-
tile composition during fruit development and ripening of ‘Alphonso’
mango. Journal of the Science of Food and Agriculture 89, 2071–2081.
Pare, P.W., and Tumlinson, J.H. 1997. De novo biosynthesis of volatiles induced
by insect herbivory in cotton plants. Plant Physiology 114, 1161–1167.
Park, Y.M., Blanpied, G.D., Jozwiak, Z., and Liu, F.W. 1993. Postharvest stud-
ies of resistance to gas diffusion in McIntosh apples. Postharvest Biology
and Technology 2, 329–339.
Paul, V., and Pandey, R. 2013. Delaying tomato fruit ripening by
1-­methylcyclopropene (1-MCP) for better postharvest management:
Current status and prospects in India. Indian Journal of Plant Physiology
18, 195–207.
Paul, V., and Pandey, R. 2014. Role of fruit’s internal gaseous microenviron-
ment on ripening: A review. Journal of Food Science and Technology 51,
1223–1250.
Paul, V., and Srivastava, G.C. 2006. Role of surface morphology in determin-
ing the ripening behaviour of tomato (Lycopersicon esculentum Mill.)
fruits. Scientia Horticulturae 110, 84–92.
Paul, V., Malik, S.K., and Srivastava, G.C. 2007. Intervarietal differences in
the surface morphology and anatomy of mango (Mangifera indica L.)
fruits. Phytomorphology 57, 211–220.
Paul, V., Pandey, R., and Srivastava, G.C. 2010a. Ripening of tomato (Solanum
lycopersicum L.). Part I. 1-Methylcyclopropene mediated delay at higher
storage temperature. Journal of Food Science and Technology 47, 519–526.
Paul, V., Pandey, R., and Srivastava, G.C. 2010b. Ripening of tomato (Solanum
lycopersicum L.). Part II. Regulation by its stem scar region. Journal of
Food Science and Technology 47, 527–533.
Paul, V., Pandey, R., and Srivastava, G.C. 2011. Tomato fruit ripening:
Regulation of ethylene production and its response. Indian Journal of
Plant Physiology 16, 117–131.

405
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Paul, V., Pandey, R., and Srivastava, G.C. 2012. The fading distinctions
between classical patterns of ripening in climacteric and non-climacteric
fruit and the ubiquity of ethylene—An overview. Journal of Food Science
and Technology 49, 1–21.
Paz, O., Jones, H.W., Prevost, B.A., and Frenkel, C. 1981. Enhancement of
fruit sensory quality by postharvest applications of acetaldehyde and
ethanol. Journal of Food Science 47, 270–274.
Pech, J.C., Bouzayen, M., and Latche, A. 2008. Climacteric fruit ripening:
Ethylene-dependent and independent regulation of ripening pathways
in melon fruit. Plant Science 175, 114–120.
Pena-Cortes, H., Barrios, P., Dorta, F., Polanco, V., Sanchez, C., Sanchez, E.,
and Ramirez, Z. 2005. Involvement of jasmonic acid and derivatives in
plant responses to pathogens and insects and in fruit ripening. Journal
of Plant Growth Regulation 23, 246–260.
Perkins-Veazie, P.M., Huber, D.J., and Brecht, J.K. 1996. In vitro growth and
ripening of strawberry fruit in the presence of ACC, STS or propylene.
Annals of Applied Biology 128, 105–116.
Peschel, S., Beyer, M., and Knoche, M. 2003. Surface characteristics of sweet
cherry fruit: Stomata number, distribution, functionality and surface
wetting. Scientia Horticulturae 97, 265–278.
Pesis, E. 1995. Induction of fruit aroma and quality by post harvest applica-
tion of natural metabolites or anaerobic conditions. In Fruit Analysis,
Modern Methods of Plant Analysis, Linsken, H.F., and Jackson, J.F. (eds.).
Vol. 18. Springer-Verlag, Berlin, 19–36.
Pesis, E. 2005. The role of the anaerobic metabolites, acetaldehyde and etha-
nol in fruit ripening, enhancement of fruit quality and fruit deteriora-
tion. Postharvest Biology and Technology 37, 1–19.
Pesis, E., and Marinansky, R. 1993. Inhibition of tomato ripening by acetalde-
hyde vapour or anaerobic conditions prior to storage. Journal of Plant
Physiology 142, 717–721.
Pesis, E., Ampunpong, C., Shusiri, B., and Hewett, E.W. 1994. Enhancement
of ethylene and CO2 production in apple fruit following short term expo-
sure to high CO2. Postharvest Biology and Technology 4, 309–317.
Pesis, E., Faiman, D., and Dori, S. 1998. Postharvest effects of acetaldehyde
vapour on ripening related enzyme activity in avocado fruit. Postharvest
Biology and Technology 13, 245–253.
Pesis, E., Levi, A., and Ben-Arie, R. 1986. Deastringency of persimmon fruits
by creating a modified atmosphere in polyethylene bags. Journal of
Food Science 51, 1014–1016.
Petro-Turza, M. 1987. Flavour of tomato and tomato products. Food Reviews
International 2, 309–351.
Podd, L.A., and van Staden, J. 1998. The role of ethanol and acetaldehyde
in flower senescence and fruit ripening—A review. Plant Growth
Regulation 26, 183–189.

406
 INTERNAL ATMOS P H ERE O F F RUITS

Podd, L.A., Hill, P.N., and van Staden, J. 2002. Physiological response and
extension of vase life of cut carnation flowers treated with ethanol and
acetaldehyde II. Protein content and enzyme activity. Plant Growth
Regulation 38, 107–117.
Polashock, J.J., Saftner, R.A., and Kramer, M. 2007. Postharvest highbush
blueberry fruit antimicrobial volatile profiles in relation to anthracnose
fruit rot resistance. Journal of the American Society for Horticultural
Science 132, 859–868.
Prasad, K., and Stadelbacher, G.J. 1974. Effect of acetaldehyde vapour on post
harvest decay and market quality of fresh strawberries. Phytopathology
64, 948–951.
Purvis, A.C., and Barmore, C.R. 1981. Involvement of ethylene in c­ hlorophyll
degradation in peel of citrus fruits. Plant Physiology 68, 854–856.
Ramayya, N., Niranjan, K., and Duncan, E. 2012. Effects of modified atmo-
sphere packaging on quality of ‘Alphonso’ mangoes. Journal of Food
Science and Technology 49, 721–728.
Ritenour, M.A., Mangrich, M.E., Beaulieu, J.C., Rab, A., and Saltveit, M.E.
1997. Ethanol effects on ripening of climacteric fruit. Postharvest Biology
and Technology 12, 35–42.
Rodoni, L., Casadei, N., Concellon, A., Alicia, A.R.C., and Vicente, A.R. 2010.
Effect of short-term ozone treatments on tomato (Solanum lycopersi-
cum L.) fruit quality and cell wall degradation. Journal of Agricultural
and Food Chemistry 58, 594–599.
Rogiers, S.Y., and Knowles, N.R. 1999. A comparison of preharvest and
postharvest ethylene production and respiration rates of saskatoon
(Amelanchier alnifolia Nutt.) fruit during development. Canadian
Journal of Botany 77, 323–332.
Rolletscheck, H., Borisjuck, L., Koschorreck, M., Wobus, U., and Weber, H.
2002. Legume embryos develop in a hypoxic environment. Journal of
Experimental Botany 53, 1099–1107.
Rupasinghe, H.P.V., Murr, D.P., Paliyath, G., and Skog, L. 2000. Inhibitory
effect of 1-MCP on ripening and superficial scald development in
‘McIntosh’ and ‘Delicious’ apples. Journal of Horticultural Science and
Biotechnology 75, 271–276.
Saltveit, M.E. 1977. Carbon dioxide, ethylene and color development in rip-
ening mature green bell peppers. Journal of the American Society for
Horticultural Science 102, 523–525.
Saltveit, M.E. 2003. Is it possible to find an optimal controlled atmosphere?
Postharvest Biology and Technology 27, 3–10.
Saltveit, M.E. 2005. Fruit ripening and fruit quality. In Tomatoes, Heuvelink, E.
(ed.). CAB International, Wallingford, UK, 145–170.
Saltveit, M.E., and Ballinger, W.E. 1983. Effect of anaerobic nitrogen and car-
bon dioxide atmosphere on ethanol production and postharvest qual-
ity of “Carlos” grapes. Journal of the American Society for Horticultural
Science 108, 462–465.

407
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Saltveit, M.E., Jr., and McFeeters, R.F. 1980. Polygalacturonase activity and
ethylene synthesis during cucumber fruit development and maturation.
Plant Physiology 66, 1019–1023.
Saltveit, M.E., Jr., and Mencarelli, F. 1988. Inhibition of ethylene synthesis
and action in ripening tomato fruit by ethanol vapours. Journal of the
American Society for Horticultural Science 113, 572–576.
Saltveit, M.E., and Sharaf, A.A. 1992. Ethanol inhibits ripening of tomato fruit
harvested at various degrees of ripeness without affecting subsequent
quality. Journal of the American Society for Horticultural Science 117,
793–798.
Salunkhe, D.K., and Do, J.Y. 1977. Bioenergetics of aroma constituents of
fruits and vegetables. Critical Reviews in Food Science and Nutrition 8,
161–190.
Sanders, M.G., and de Wild, H.P.J. 2003. The relation between in vivo eth-
ylene production and oxygen partial pressure. Postharvest Biology and
Technology 30, 143–151.
Saniewski, M., and Czapski, J. 1985. Stimulatory effect of methyl jasmonate
on the ethylene production in tomato fruits. Experientia 41, 256–257.
Sasaki, K., Saito, T., Lamsa, M., Caldentey, K.M.O., Suzuki, M., Ohyama, K.,
Muranaka, T., Ohara, K., and Yazaki, K. 2007. Plants utilize isoprene
emission as a thermotolerance mechanism. Plant and Cell Physiology
48, 1254–1262.
Sawamura, M., Knegt, E., and Bruinsma, J. 1978. Levels of endogenous eth-
ylene, carbon dioxide, and soluble pectin and activities of pectin methyl­
esterase and polygalacturonase in ripening tomato fruits. Plant and Cell
Physiology 19, 1061–1069.
Schotsmans, S.W., Verlinden, B.E., Lammertyn, J., and Nicolai, B.M. 2004.
The relationship between gas transport properties and histology of
apple. Journal of the Science of Food and Agriculture 84, 1131–1140.
Scully, A.D., and Horsham, M.A. 2008. Active packaging for fruits and veg-
etables. In Intelligent and Active Packaging for Fruits and Vegetables,
Wilson, C.L. (ed.). CRC Press, Taylor & Francis Group, Boca Raton,
FL, 57–71.
Shellie, K.C., and Saltveit, M.E., Jr. 1993. The lack of a respiratory rise in
muskmelon fruit ripening on the plant challenges the definition of cli-
macteric behaviour. Journal of Experimental Botany 44, 1403–1406.
Shi, J.X., Goldschmidt, E.E., Goren, R., and Porat, R. 2007. Molecular, bio-
chemical and anatomical factors governing ethanol fermentation metab-
olism and accumulation of off-flavours in mandarins and grapefruit.
Postharvest Biology and Technology 46, 242–251.
Shi, J.X., Pora, R., Goren, R., and Goldschmidt, E.E. 2005. Physiological
responses of ‘Murcott’ mandarins and ‘Star Ruby’ grapefruits to anaero-
bic stress conditions and their relation to fruit taste, quality, and emission
of off-flavour volatiles. Postharvest Biology and Technology 38, 99–105.
Shiomi, S., Yamamoto, M., Nakamura, R., and Inaba, A. 1999. Expression of
ACC synthase and ACC oxidase genes in melons harvested at different

408
 INTERNAL ATMOS P H ERE O F F RUITS

stage of maturity. Journal of the Japanese Socety for Horticultural Science

68, 10–17.
Shipway, M.R., and Bramlage, W.J. 1973. The effect of carbon dioxide on
activity of apple mitochondria. Plant Physiology 51, 1095–1098.
Smith, S., Geeson, J., and Stow, J. 1987. Production of modified atmospheres
in deciduous fruits by the use of films and coatings. HortScience 22,
772–777.
Solomos, T. 1987. Principles of gas exchange in bulky plant tissue. HortScience
22, 766–771.
Sornsrivichai, J., Yantarasri, T., and Gemma, H. 1998. Respiration kinetics of
mangoes and Asian pear fruits under MA conditions and its relation to
the ripening behaviour. Acta Horticulturae 464, 339–344.
Staudt, M., and Bertin, N. 1998. Light and temperature dependence of the
emission of cyclic and acyclic monoterpenes from holk oak (Quercus
ilex L.) leaves. Plant and Cell Environment 21, 85–395.
Steindel, F., Beauchamp, J., Hansel, A., Kesselmeier, J., Kleist, E., Kuhn, U.,
Wisthaler, A., and Wildt, J. 2005. Stress induced VOC emissions from
mildew infested oak. Geophysics Research Abstract 7, EGU05-A-03010.
Stepanova, A.N., and Alonso, J.M. 2005. Ethylene signaling and response
pathway: A unique signaling cascade with a multitude of inputs and out-
puts. Physiologia Plantarum 123, 195–206.
Tadesse, T., Nichols, M.A., and Hewett, E.W. 1998. Ripening of attached and
detached sweet pepper fruit cv. ‘Domino’. Acta Horticulturae 464, 503.
Tandon, K.S., Baldwin, E.A., and Shewfelt, R.L. 2000. Aroma perception of
individual volatile compounds in fresh tomatoes (Lycopersicon esculen-
tum Mill.) as affected by the medium of evaluation. Postharvest Biology
and Technology 20, 261–268.
Tandon, K.S., Jordan, M., Goodner, K.L., and Baldwin, E.A. 2001.
Characterization of fresh tomato aroma volatiles using GC-olfactometry.
Proceedings of the Florida State Horticulture Society 114, 142–144.
Thompson, A.K. 2003. Postharvest technology of fruits and vegetables. In
Fruits and Vegetables: Harvesting Handling and Storage, Thompson,
A.K. (ed.). Blackwell, Oxford, 354.
Tian, M.S., Hewett, E.W., and Lill, R.E. 1994. Effects of carbon dioxide on eth-
ylene forming enzyme in Japanese pear and apple. Postharvest Biology
and Technology 4, 1–12.
Toivonen, P.M.A. 1997. Non-ethylene, non-respiratory volatiles in harvested
fruits and vegetables: Their occurrence, biological activity and control.
Postharvest Biology and Technology 12, 109–125.
Toivonen, P.M.A., Escobar, S., and Emona, J.P. 1999. Response of three
raspberry cultivars to different modified atmosphere conditions. Acta
Horticulturae 505, 33–38.
Trainotti, L., Pavanello, A., and Casadoro, G. 2005. Different ethylene receptors
show an increased expression during the ripening of strawberries: Does
such an increment imply a role for ethylene in the ripening of these non-
climacteric fruits? Journal of Experimental Botany 56, 2037–2046.

409
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Tucker, G.A. 1993. Introduction. In Biochemistry of Fruit Ripening, Seymour,

G., Talor, J., and Tucker, G. (eds.). Chapman & Hall, London, 1–51.
Utto, W., Mawson, A.J., and Bronlund, J.E. 2008. Hexanal reduces infection
of tomatoes by Botrytis cinerea whilst maintaining quality. Postharvest
Biology and Technology 47, 434–437.
Uys, D.C. 1974. Keeping Quality of Grapes with Special Reference to Skin
Characteristics. Department of Agricultural Technical Services,
Agricultural Research, South Africa.
Van Loon, L.C., Geraats, B.P.J., and Linthorst, H.J.M. 2006. Ethylene as a
modulator of disease resistance in plants. Trends in Plant Science 11,
185–191.
Varoquaux, P., and Ozdemir, I.S. 2005. Packaging and produce degradation.
In Produce Degradation Pathways and Prevention, Lamikanra, O., Imam,
S., and Ukuku, D. (eds.). CRC Press, Taylor & Francis Group, Boca
Raton, FL, 117–153.
Verboven, P., Kerckhofs, G., Mebatsion, H.K., Ho, Q.T., Temst, K., Wevers, M.,
Cloetens, P., and Nicolai, B.M. 2008. Three-dimensional gas exchange
pathways in pome fruit characterized by synchrotron x-ray computed
tomography. Plant Physiology 147, 518–527.
Villavicencio, L.E., Blankenship, S.M., Sanders, D.C., and Swallow, W.H.
2001. Ethylene and carbon dioxide concentrations in attached fruits of
pepper cultivars during ripening. Scientia Horticulturae 91, 17–24.
Wang, H., Liang, X., Huang, J., Zhang, D., Lu, H., Liu, Z., and Bi, Y. 2010.
Involvement of ethylene and hydrogen peroxide in induction of alterna-
tive respiratory pathway in salt-treated Arabidopsis calluses. Plant and
Cell Physiology 51, 1754–1765.
Wann, E.V. 1996. Physical characteristics of mature green and ripe tomato
fruit tissue of normal and firm genotypes. Journal of the American
Society for Horticultural Science 121, 380–383.
Wardlaw, C.W., and Leonard, E.R. 1936. Studies in tropical fruits. Annals of
Botany 50, 622–653.
Wasternack, C. 2007. Jasmonates: An update on biosynthesis, signal trans-
duction and action in plant stress response, growth and development.
Annals of Botany 100, 681–697.
Watkins, C.B., Barden, C.L., and Bramlage, W.J. 1993. Relationship between
alpha-farnesene, ethylene production and superficial scald develop-
ment of apples. Acta Horticulturae 343, 155–160.
Wechter, W.P., Levi, A., Harris, K.R., Davis, A.R., Fei, Z., Katzir, N., Giovannoni,
J.J., Salman-Minkov, A., Hernandez, A., Thimmapuram, J., Tadmor, Y.,
Portnoy, V., and Trebitsh, T. 2008. Gene expression in developing water-
melon fruit. BMC Genomics 9, 275.
Weichmann, J. 1986. The effect of controlled atmosphere storage on the
sensory and nutritional quality of fruits and vegetables. Horticultural
Reviews 8, 101–127.

410
 INTERNAL ATMOS P H ERE O F F RUITS

Wills, R.B.H., and Ku, V.V.V. 2002. Use of 1-MCP to extend the time to ripen
the green mature tomatoes and postharvest life of ripe tomatoes.
Postharvest Biology and Technology 26, 86–90.
Wills, R.B.H., Ku, V.V.V., and Leshem, Y.Y. 2000. Fumigation with nitric oxide
to extend the postharvest life of strawberries. Postharvest Biology and
Technology 18, 75–79.
Xu, L. 1999. Use of ozone to improve the safety of fresh fruits and vegetables.
Food Technology 53, 58–61.
Xu, Z.C., Ikoma, Y., Yano, M., Ogawa, K., and Hyodo, H. 1998. Varietal differ-
ences in the potential to produce ethylene and gene expression of ACC-
synthase and ACC-oxidase between ‘Kuimi’ and ‘Hong Xin’ of Chinese
kiwifruit. Journal of the Japanese Society for Horticultural Science 67,
204–209.
Yahia, E.M. 2009. Introduction. In Modified and Controlled Atmospheres for the
Storage, Transportation and Packaging of Horticultural Commodities,
Yahia, E.M. (ed.). CRC Press, Taylor & Francis Group, Boca Raton, FL,
1–16.
Yang, C.X., and Shewfelt, R.L. 1999. Effects of sealing of stem scar on ripen-
ing rate and internal ethylene, oxygen and carbon dioxide concentra-
tions of tomato fruits. Acta Horticulturae 485, 399–404.
Yang, S.F., and Hoffman, N.E. 1984. Ethylene biosynthesis and its regulation
in higher plants. Annual Review of Plant Physiology 35, 155–189.
Yang, S.F., and Pratt, H.K. 1978. The physiology of ethylene in wounded plant
tissue. In Biochemistry of Wounded Plant Tissue, Kau, G. (ed.). Berlin,
Walter de Gruyter, 595–622.
Yip, W.K., Jiao, X.Z., and Yang, S.F. 1988. Dependence of in vitro ethylene
production rate on 1-aminocyclopropane-1-carboxylic acid content and
oxygen concentrations. Plant Physiology 88, 553–558.
Yoo, S.D., Cho, Y., and Sheen, J. 2009. Emerging connections in the ethylene
signaling network. Trends in Plant Science 14, 270–279.
Zagory, D., and Kader, A.A. 1988. Modified atmosphere packaging of fresh
produce. Food Technology 42, 70–77.
Zambre, S.S., Venkatesh, K.V., and Shah, M.G. 2010. Tomatoes redness
for assessing ozone treatment to extend the shelf life. Journal of Food
Engineering 96, 463–468.
Zhang, Z., Huber, D.J., Hurr, B.M., and Rao, J. 2009. Delay of tomato fruit
ripening in response to 1-methylcyclopropene is influenced by internal
ethylene levels. Postharvest Biology and Technology 54, 1–8.
Ziosi, V., Bregoli, A.M., Bonghi, C., Rasori, A., Biondi, S., Costa, G., and
Torrigiani, P. 2007. Jasmonates delay ripening by interfering with eth-
ylene biosynthesis and perception and with polyamine accumulation
in peach fruit. In Advances in Plant Ethylene Research, Ramina, A.,
Chang, C., Giovannoni, J., Klee, H., Perata, P., and Wolterinig, E. (eds.).
Springer, Wageningen, the Netherlands, 109–110.

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Chapter 13

Proteomics of
Fruit Development
and Ripening
Aloysius Wong,1 Ludivine Thomas,1
Christoph Gehring,1 and Claudius Marondedze2
1King Abdullah University of Science and Technology,
Thuwal, Kingdom of Saudi Arabia
2University of Cambridge, Cambridge, United Kingdom

Abstract414
13.1 Introduction 414
13.2 Characteristics of Climacteric and Nonclimacteric Fruit
Ripening416
13.3 Proteomic Tools and Approaches for Fruit Ripening
Assessment417
13.3.1  Challenges in Proteomics Analysis 418
13.3.2 Protein Sample Preparation: Tissue Disruption,
Homogenization, and Solubilization 418
13.3.3 Protein Separation: ­Gel-Based and Gel-Free
Technologies421
13.4  Proteomes of Climacteric and Nonclimacteric Fruits 422
13.4.1 Overrepresented Functional Categories
during Development and Ripening 422

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13.5  Pathway Analysis Using KEGG 426

13.5.1 Proteins Associated with Carbohydrate
Metabolism426
13.5.2  Proteins Associated with Energy Metabolism 431
13.5.2.1 Proteins with a Role in Carbon
Fixation431
13.5.2.2 Proteins Associated with the
Pentose Phosphate, Glycolysis,
and Pyruvate Metabolisms 432
13.5.2.3  Proteins Involved in the TCA Cycle 436
13.5.3 Proteins Involved in Amino Acid Metabolism
and Ethylene Biosynthesis 436
13.5.4  Proteins Involved in Flavonoid Biosynthesis 437
13.6  Conclusions and Future Perspectives 438
References439

Abstract
Comparative proteomics has emerged as a powerful tool used to study com-
plex biological processes in fruits throughout their development and ripen-
ing. This is greatly aided by the rapid growth of genomics, transcriptomics,
and expressed sequence tag (EST) databases, which allow for protein iden-
tification and pave the way for systems analyses and inference of molecular
data. Fruit development and ripening are complex developmental processes
that involve well-coordinated biological programs, the knowledge of which
has valuable economic ramifications centered on the agricultural industry.
Besides, fruit ripening is accompanied by numerous phenotypic and physi-
ological changes, such as skin and hypanthium color and increase in sugar
levels, which are regulated by environmental factors such as light and tem-
perature and internal factors such as hormonal and gene regulation. The lat-
ter is key in the classification of fruit species in their respective climacteric
and nonclimacteric categories. Comparative proteomics is therefore a useful
tool to gain information on the molecular events taking place during fruit
maturation, in addition to finding biotechnological strategies to improve hor-
ticultural traits such as fruit quality, shelf life, and yield. In this chapter, an
overview of methods utilized in fruit proteomics, as well as a global proteome
and systems biology analysis of fruits during ripening, is presented.

13.1 Introduction
Fruit development is a highly coordinated process that involves irreversible
cellular events and is accompanied by systematic molecular and physiological

414
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changes (Bonghi and Manganaris, 2012). On the other hand, fruit ripening
correlates with a series of complex developmental changes, usually involving
various physiological processes, such as chlorophyll degradation, pigment
biosynthesis (carotenoid and xanthophyll) (Grassi et al., 2013), tissue soften-
ing (Payasi et al., 2009), starch degradation, simple sugar accumulation (Li
et  al., 2012), and volatile compounds and organic acid production (Nergiz
and Ergönül, 2009; Etienne et al., 2013; Rambla et al., 2014). The final stage
of fruit development usually manifests apparent color, flavor, aroma, and
texture changes that are appealing to consumers and also to the broader
agricultural industry. In 2012, global banana (Musa acuminata) production
exceeded 100 million metric tons, while apples (Malus × domestica), oranges
(Citrus × sinensis), and grapes (Vitis vinifera) combined reached more than
200  ­million metric tons (FAOSTAT, 2013). Due to the growing economic
impact of fruit crops, understanding of events that occur during fruit mat-
uration has become of paramount importance, as this knowledge can be
exploited for breeding strategies to achieve desirable traits and improved
yield. While postharvest innovations such as fruit handling and storage strat-
egies have improved tremendously (Palma et  al., 2011), physiological and
intracellular events, such as the expression and regulation of genes, signal-
ing mechanisms, and the biosynthesis and regulation of key molecules and
enzymes responsible for the fruit ripening process, have just recently gained
momentum. Better understanding of fruit maturation at the molecular level
is vital for further progress of postharvest technologies. In order to address
these challenges, advanced and high-throughput technologies have been
applied to discover the gene expression profile (Fei et al., 2004), the biosyn-
thesis of metabolites (Alba et al., 2005) and other molecules unique to fruit
development, and the proteome changes of various fruits (Sarry et al., 2004;
Rocco et al., 2006). The latter has recently seen a “burst” in the fruit science
field as reflected by more than 50 scientific literature outputs in the last 5
years. This is mainly due to the increasing availability of fruit crop genomes
and transcriptomes, from which plant biologists can extract functional infor-
mation. Since the fruit proteome is dynamic, elucidation of its profile at a
given development stage or in response to various signaling molecules or
stimuli is required to understand and capture key molecular events, which
are either not possible or less accurate with genome analyses alone. One
example is the occurrence and regulation of posttranslational modifications
(PTMs) that influence protein functions and interactions, and occur down-
stream and independently of the genetic code (Marondedze et  al., 2014).
These PTMs render functional predictions from the genetic sequence inac-
curate. Additionally, recent advancements in proteomic approaches, such
as protein sample preparation, separation methods, and identification and
visualization methods (for review, see Orellana and Nilo, 2012), have aided
comparative analyses of the fruit proteome. This chapter highlights meth-
odological advances in fruit development and ripening proteomics; offers

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fresh views, analyses, and interpretations of proteome profiles gathered from

­economically important climacteric and nonclimacteric fruits; and discusses
the challenges and future directions in fruit proteomics.

13.2  Characteristics of Climacteric and

Nonclimacteric Fruit Ripening
Changes that occur at both the molecular and physiological levels are the
determinant factors for fruit classification, and as such, they can be catego-
rized as climacteric or nonclimacteric based on how they achieve ripening.
Climacteric fruits are characterized by a sudden and rapid increase of the
phytohormone ethylene and elevation of the respiration rate during ripening
(Adams-Phillips et al., 2004). Examples of fruits demonstrating this character-
istic include apple, banana, pear, plum, peach, melon, and tomato. Additionally,
this gaseous molecule is responsible for tissue softening, which decreases
the flesh firmness of fruits as they ripen (Hiwasa et al., 2003; Marondedze
and Thomas, 2012b). On the other hand, nonclimacteric fruits ripen indepen-
dently of ethylene concentrations; that is, they do not exhibit drastic changes
in ethylene biosynthesis (Adams-Phillips et al., 2004), as observed for orange,
lemon, grape, strawberry, and pepper. While the “­ethylene peak” is a clear
distinction of climacteric and nonclimacteric ripening of fruits, other physi-
ological characteristics, such as texture, ­flavor, color, and even metabolic
events, are undistinguishable (Giovannoni, 2001). As noted by Palma et  al.
(2011), two fruits of the same family, such as tomato and pepper, can display
contrasting ripening behavior; thus, the ethylene profile of climacteric and
nonclimacteric fruits is not phylogeny specific (Palma et al., 2011).
Due to the economic impact of fleshy fruits, the molecular events of
ethylene, such as the biosynthesis, the signaling pathways, and the regula-
tion of gene expressions governing fruit texture, color, taste, and aroma, have
been a subject of extensive research. In the last decades, implementation of
this knowledge allowed for progress in retarding the physiological events
related to postripening of fruits, thus reducing waste and the concomitant
economic repercussions of these losses. Through genomics, these molecular
mechanisms have been elucidated and extensively reviewed by Bapat et al.
(2010), who not only described ethylene biosynthesis, signal transduction,
and the regulation of target genes downstream of ethylene perception in the
tomato model system, but also provided evidence from other economically
important fruits, such as apple, avocado, banana, grape, kiwi, mango, melon,
papaya, pear, pineapple, and strawberry. Given the importance of ethylene in
the ripening of climacteric fruits, here, we briefly review the biotechnologi-
cal implications of this knowledge in tomato and other fruits.
Tomato was selected as a fruit model system to elucidate the molecu-
lar events of ethylene and for the generation of transgenic plants mainly

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Proteomics of Fruit Development and Ripening

because of its short life cycle and the availability of various tomato genomic
resources, such as microarrays, expressed sequence tags (ESTs), and the
recently completed tomato genome (Tomato Genome Consortium, 2012;
Kimura and Sinha, 2008). In addition to the development of effective and
stable tomato transformation systems (Ruf et al., 2001), numerous mutant
lines have also been generated and used for the characterization of ethyl-
ene biosynthesis and regulation (Saito et al., 2011). Several studies on key
genes involved in ethylene biosynthesis, including S-adenosylmethionine
decarboxylase, 1-aminocyclopropane-1-carboxylic acid synthase (ACS), and
1-aminocyclopropane-1-carboxylic acid oxidase (ACO), revealed that a reduc-
tion in ethylene synthesis in tomato fruit can delay fruit ripening, thus
increasing fruit shelf life (Hamilton et al., 1990; Oeller et al., 1991; Good
et  al., 1994; Lokkamlue and Huehne, 2013). Further, transgenic tomato
lines with reduced ethylene perception, such as those involving the Never-
ripe and the Ethylene receptor (ETR) genes, also achieved delay in fruit rip-
ening (Wilkinson et al., 1997; Ciardi et al., 2000).
While limited, similar evidence in other fruits, such as apple, melon,
and papaya, has also been reported, of which some displayed encouraging
fruit ripening alterations (Flores et al., 2001; Dandekar et al., 2004; Lopez-
Gomez et al., 2009). In apple fruits, ACS or ACO genes were silenced and
fruits of the transgenic plants showed a firmer texture and increased shelf
life, although biochemical events such as sugar or acid accumulation were
unchanged (Dandekar et  al., 2004). In ACO-silenced melon plants, the
chlorophyll and carotenoid pigments remained intact in the rind, and other
biochemical contents (sucrose and citric acid) were accumulated in the
pulp (Flores et al., 2001). Cosuppression of ACO in the tropical fruit papaya
also achieved delayed ripening and alterations in CO2 production and color
development (Lopez-Gomez et  al., 2009). It is noted that the suppression
of ethylene perception approach, similar to the ETR suppression studies
in transgenic tomato plants, has yet to be reported in other fruits. As more
genetic resources and tools become available for other economically impor-
tant fruits, current discoveries on ripening from the tomato model system
can be translated to these fruits.

13.3  Proteomic Tools and Approaches for

Fruit Ripening Assessment
While the gene and hormonal regulations of ripening can be contrasted
between climacteric and nonclimacteric fruits, the phenotypic characteris-
tics of ripening fruits are often very similar. Additionally, genetic expression
data often correlate poorly with the manifested traits. Therefore, in order
to uncover the underlying molecular events that occur during ripening,
proteomic studies should be attempted since protein populations in fruit

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tissues expressed at a given time under specific conditions are diagnostic

of fruit characteristics that manifest as observable traits, such as texture,
aroma, taste, and color. The ability to detect qualitative and quantitative pro-
tein changes across fruit development and ripening can reveal the underly-
ing regulatory networks and metabolic events that form a direct correlation
between the genomics profile and phenotypic characteristics. To achieve
this, various experimental tools and approaches have been developed, and
here, we highlight some common and currently applied techniques in fruit
proteomics for the systemic analysis of protein expression (Figure 13.1).

13.3.1  Challenges in Proteomics Analysis

In contrast to genomics and transcriptomics analyses, a systemic evaluation
of the proteome, and in particular the plant proteome, is more cumbersome.
Indeed, protein complexity, as reflected by their physical and chemical het-
erogeneity, as well as their structural conformations, translates to the need
for careful optimization of a number of experimental steps, including extrac-
tion, homogenization, fractionation, and detection. As such, mapping and
monitoring of the entire proteome of an entire tissue, and even a single cell,
remain unachievable. While a global assessment of gene expression is becom-
ing more systematic and less costly with the availability of high-throughput
methods such as next-generation sequencing (Heller, 2002; Metzker, 2010),
mRNA levels have been shown to poorly correlate to the abundance of their
protein products (Gygi et al., 1999; Greenbaum et al., 2003; Laurent et al.,
2010). This is largely due to posttranslational modifications (PTMs), such
as the attachment or removal of carbohydrates, lipids, or amino acids, which
alters the chemical and structural properties of proteins. These PTMs result
in increased functional diversity of the proteome, while no changes in gene
sequence and sometimes expression level are observed. However, PTM
detection remains a challenge due to their low stoichiometry and the meth-
ods of sample preparation and analyses used, which often result in their loss
(Mann and Jensen, 2003). Despite technical advancements in genome-scale
studies and difficulties in proteomic approaches, protein expression profiles
are desirable not only to complement transcriptomics data, but also to facili-
tate linking the genome to the resulting phenotypes and the environment.

13.3.2  Protein Sample Preparation: Tissue Disruption,

Homogenization, and Solubilization
The first step, and arguably the most crucial, for the recovery of proteins
from fruit tissues is sample preparation (Giavalisco et al., 2003). In addi-
tion to the presence of a cell wall (Rose et al., 2004), fruit cells have a high

418
Proteomics of Fruit Development and Ripening

Tissues disruption

Fruit tissues
Homogenization
Protein extract

Quantitation
Gel-based (2-DE) Gel-free
Protein separation

pH gradient
IEF Alkylation, reduction
and digestion by trypsin
Alkylation, reduction and with/without labels

SDS-PAGE MW Labeled or un-labeled

large
LC-based
Visualization
separation of
peptides

Small

MS and MS/MS
Spot excision and in-gel Data Analysis
tryptic digestion

Detection and identification

Figure 13.1  A diagrammatic overview of proteomics experimental ­workflows.

Fruit tissues are first disrupted using mechanical methods such as liquid ­nitrogen–
assisted mortar and pestle grinding or sonication, before homogenization in
TCA–acetone or phenol-containing buffers. The solubilized proteins are then
separated using either a gel-based (2-DE) or gel-free approach. In the gel-
based approach, proteins, labeled (e.g., with fluorescent dyes) or unlabeled,
are separated by charge Isoelectric focusing (IEF) in the first dimension and then
by size in the second dimension. The protein spots can be visualized, excised,
and trypsin digested before peptide identification and quantification by tandem
mass spectrometry. In the gel-free approach, proteins, labeled (e.g., TMT and
iTRAQ) or unlabeled, are first digested by trypsin before separation by either
OFFGEL or liquid chromatography. Separated peptide fractions are then identi-
fied and quantified by tandem mass spectrometry. The resulting protein list is
then analyzed using various bioinformatics tools and software packages.

water content and low protein concentration (Saravanan and Rose, 2004).
Protein extraction is further complicated by the presence of relatively
high amounts of contaminants, such as acids, lipids, polysaccharides, and
phenols, which interfere with downstream analyses. Taking into consid-
eration these challenges, the sample preparation strategy should include

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optimized tissue disruption, homogenization, and protein solubilization.

Different methods or combinations of methods can be applied, depending
on the type of fruits and their degree of firmness, to achieve as finely pow-
dered tissues as possible and a maximum protein yield. Tissue grinding
of frozen fruit tissues in liquid nitrogen using a mortar and pestle is by far
the most common method for tissue disruption. In some cases, the use of
grinding aids, such as fine beads made of glass, sand, or alumina, improves
tissue disruption by shearing of the cell wall during mechanical grinding,
thanks to increased abrasion of the rough edges of beads against the cell
wall (Giavalisco et al., 2003). Conversely, for fruits that have firmer fibrous
tissues or unripe fruits, more vigorous tissue disintegration methods,
using instruments such as high-speed blenders equipped with specially
designed blades, before or after the grinding, can be employed (Vincent
et  al., 2006; Marondedze et  al., 2014). Other reported tissue disruption
technologies include sonication and acoustics. The former was used in
tandem with liquid nitrogen for assisted grinding of capsicum and grape
berry tissues (Lee et al., 2006; Di Carli et al., 2011), while the latter offers
a contact-free tissue disruption method to increase extraction efficiency,
especially in a large-scale setting, and has been tried in soybean and rice
(Toorchi et al., 2008).
Following cell disruption and homogenization, protein solubilization
is attempted with the aim of recovering the maximum amount of protein
while limiting contamination from other compounds and interfering arti-
facts (Shaw and Riederer, 2003). It must be noted that since proteins have
very diverse chemical and physical properties (e.g., charge, size, and hydro-
phobicity), no single solubilization method or solvent system can recover an
entire given proteome. As such, various methods have been explored and
optimized in order to capture the maximum amount of proteins. The use of
two methods, trichloroacetic acid (TCA) in acetone and phenol-based pro-
tein precipitation, has been frequently reported in the literature (Isaacson
et al., 2006; Marondedze, 2011; Marondedze and Thomas, 2012a; Wu et al.,
2014b). Protein precipitation by TCA and acetone increases protein concen-
tration while removing contaminants such as lipids, cell wall components,
and phenolic compounds that might interfere with downstream separation
of proteins (X. Wu et al., 2014a). This is particularly useful when using tis-
sues from older plants or that contain high levels of phenols and saccha-
rides. The phenol extraction of proteins method described by Hurkman
and Tanaka (1986) is also commonly employed, although it is more time-
consuming than the TCA in acetone. The method has notably been used
in apple, potato, and banana (Carpentier et al., 2005). A sequential precipi-
tation using first phenol and then TCA in acetone can remove pigments
and lipids, as documented with mature grape berries (Wang et al., 2003;
Vincent et al., 2006). In another study, protein precipitation first in p
­ henol,
followed by methanol and ammonium acetate precipitation, yielded a

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Proteomics of Fruit Development and Ripening

protein mixture with minimal contaminants from recalcitrant tissues, such

as olive leaves (Wang et al., 2003) and wood (Vander Mijnsbrugge et al.,
2000). This approach can also be applied to fruit tissues. The addition of
reducing and chelating agents, such as polyvinylpolypyrrolidone (PVPP)
in the extraction buffer of the phenol-based method, was also reported as
a means to further reduce contaminants (Isaacson et al., 2006; Martinez-
Esteso et al., 2011; Marondedze et al., 2014). While both the TCA in acetone
and phenol-based extraction methods are widely used in fruit proteomics,
direct comparisons between the two are limited. Studies conducted by
Marondedze and Thomas (2012b) and Zheng et  al. (2007) on apple and
strawberry fruits revealed that both methods yielded substantial qualita-
tive and quantitative differences. In these two studies, the phenol-based
method specifically allowed for recovery of a greater number of proteins.
Although the phenol-based extraction method generally outperforms TCA
in acetone, especially for recalcitrant fruit tissues, a substantial number
of unique proteins can be recovered from both techniques. It is probably
worth envisaging a two-step extraction procedure, first performing a TCA
in acetone precipitation, followed by a second step with phenol to capture a
maximum amount of proteins, while concurrently removing contaminants
such as lipids and pigments (X. Wu et al., 2014b).

13.3.3  Protein Separation: ­Gel-Based

and Gel-Free Technologies
Protein populations can then be analyzed using either gel-based or gel-free
techniques. Due to the complexity and characteristics of the fruit proteome,
most fruit proteomic studies to date used two-dimensional gel electropho-
resis (2-DE) for the separation of protein samples. For example, this tech-
nique has been used in citrus (Muccilli et  al., 2009), date (Marondedze
et  al., 2014), capsicum (Aizat et  al., 2013), mango (Andrade et  al., 2012),
apple (Marondedze and Thomas, 2012b), papaya (Nogueira et  al., 2012),
tomato (Xu et al., 2013), pear (Pedreschi et al., 2007), and grape (Giribaldi
et al., 2007) prior to tryptic digestion and mass spectrometry (MS) analysis
for protein identification. Several fruit proteomic studies also reported using
a combination of 2-DE and fluorescent dyes for visualization of proteins
(Riederer, 2008; Gauci et  al., 2011; Marondedze et  al., 2014). Despite the
powerful resolving ability of 2-DE, certain classes of proteins, in particular
those with high hydrophobicity, are notoriously difficult to separate with
this technology (Santoni et al., 1999, 2000). Even in optimum conditions,
2-DE contains an average of 1000–2000 unique protein spots (Rose et al.,
2004), although some reported up to 10,000 distinct spots (Hatzimanikatis
et al., 1999; Klose, 1999), representing as little as 5% of the total proteome
(Heazlewood and Millar, 2003).

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Gel-free approaches have gained popularity in recent years due to

the drawbacks of 2-DE, especially when dealing with membrane proteins.
Generally, proteomes are first digested to peptides, fractionated by liquid
chromatography (LC), and identified by tandem mass spectrometry (MS/
MS). Gel-free strategies can also be quantitative by labeling peptides using
either isobaric tags for relative and absolute quantification (iTRAQ) or tan-
dem mass tags (TMTs), or by doing label-free quantitation, such as spec-
tral counting (Old et al., 2005) or ion intensity measurement (Silva et al.,
2005). To the best of our knowledge, the label-free quantitative proteomics
approach was documented three times for fruits: in the metabolic analysis
of citrus fruit development (Katz et al., 2011) and in the proteome analyses
of both mango (H.X. Wu et al., 2014) and banana (Esteve et al., 2013).

13.4  Proteomes of Climacteric and

Nonclimacteric Fruits
After just more than a decade, the proteomics of fruit development and rip-
ening on economically valuable plants is still in its infancy; however, it pro-
vides a wide range of novel insights pertaining to developmental regulation
mechanisms (Molassiotis et al., 2013). Here, a gene ontology (GO) analysis
was carried out using fruit proteomics data gathered from public reposito-
ries and further assigned to pathways using the Kyoto Encyclopedia of Genes
and Genomes (KEGG). For climacteric fruits, data were obtained from apple
(Marondedze and Thomas, 2012b), apricot (D’Ambrosio et al., 2013), banana
(Toledo et al., 2012), date (Marondedze et al., 2014), mango (Andrade et al.,
2012), papaya (Nogueira et  al., 2012), and peach (Prinsi et  al., 2011), and
for nonclimacteric fruits, data from grape (Martinez-Esteso et  al., 2011;
Kambiranda et al., 2014), olive (Katz et al., 2011; Bianco et al., 2013), orange
(Katz et al., 2010, 2011), and strawberry (Bianco et al., 2009) were collected.
Functional classification based on the pathway analysis and the number of
proteins targeted by climacteric and nonclimacteric ripening processes was
performed using Blast2GO v1.0 (Conesa et al., 2005; Conesa and Gotz, 2008;
Gotz et al., 2008, 2011). A total of 875 differentially expressed protein entries
from climacteric species and 1056 differentially expressed protein entries
from nonclimacteric species were used for the GO analysis.

13.4.1  Overrepresented Functional Categories

during Development and Ripening
Fruit proteomic data from climacteric and nonclimacteric plant spe-
cies were gathered and subjected to gene ontology (GO) analysis using
Blast2GO (Conesa et al., 2005; Conesa and Gotz, 2008; Gotz et al., 2008,

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2011) to obtain biological and molecular information that reflect the events
leading to fruit ripening. Based on our analysis, the major portion (21%)
of the differentially expressed proteins in climacteric species were plastid
proteins, while in nonclimacteric species, both plastid and plasma mem-
brane proteins shared a reduced majority of 17% (Figure 13.2A). The high
representation of proteins from plastids during fruit development and ripen-
ing is partly attributed to the biosynthesis of pigments such as carotenoids,
which give fruits their distinctive colors. While fruit plastids have reduced
photosynthetic capability in comparison to leaves, metabolic activities such
as the catabolism of starch and metabolites for the bioaccumulation of pig-
ments, lipids, and amino acids are high (Bouvier and Camara, 2006). Plasma
membrane proteins are also overrepresented in both climacteric (13%) and
nonclimacteric (17%) species since new membranes are being developed
to sequester pigments and lipid derivatives synthesized during ripening
(Camara et al., 1995). A remodeling of membrane protein composition and
organization takes place during the shift from photosynthetic green fruit to
less or nonphotosynthetic colored fruits during the transition from develop-
ment to ripening, and this possibly explains the overrepresentation of the
plasma membrane category. Key members of the membrane category, and
specifically in the plastid, include translocators, which function to shuttle
sugars and metabolites to and from the cytosol (Flügge, 1999). The phos-
phate translocators, involved in recruiting carbon source hexose phosphate
from the cytosol, and polyphenol oxidases, which are responsible for fruit
browning, are located at the plasma and thylakoid membranes and were
detected as overexpressed during fruit ripening (Flügge, 1999; Mayer,
2006). In climacteric fruits, overrepresented proteins from the cytosol rep-
resented 11% of total differentially expressed proteins. Contributions from
differentially expressed proteins from the extracellular region, mitochon-
dria, and nucleus were comparable between climacteric and nonclimacteric
species. However, a lower ratio of nucleus proteins was detected in noncli-
macteric (4%) than in climacteric (10%) species. Transcription factors, such
as the MADS box transcription factor ripening inhibitor (RIN), are known
to accumulate in the nucleus during ripening, and their absence has been
shown to halt fruit ripening (Ito et al., 2008). The higher representation of
nuclear proteins in climacteric species could be attributed to the necessity
for ethylene-related transcription factors, such as the ethylene-insensitive
(EIN) proteins, for the ripening of climacteric fruits (Chaves and de Mello-
Farias, 2006). Interestingly, Golgi apparatus proteins contributed to 5% of
overrepresented proteins in nonclimacteric species, although this category
was absent in climacteric species.
With regard to the molecular function, similar ratios were detected in
climacteric and nonclimacteric species for the nucleotide binding (31% and
30%, respectively), hydrolase activity (24% and 21%, respectively), transfer-
ase activity (18% and 15%, respectively), and protein binding (17% and 16%,

423
P OST H AR V EST RI P ENING P H Y SIOLOG Y

5 Plastid Cell wall

6
21
Plasma membrane Golgi apparatus
6 4 3 17
5
Cytosol Thylakoid
5
7 7 Mitochondria Nucleus
A
Cellular component 17 Vacuole Non-membrane
8 (%) bound organelle
13 Extracellular region
10
8

11 15
10 11
10

Nucleotide binding
10

7 Hydrolase activity
7 31 Transferase activity
17 30
5
Protein binding
B
Molecular function RNA binding
16 (%)
Structural molecule activity

Transporter activity
18 15 21

24

21 Metabolic process Multicellular organismal process

3 2
3
4 2 22 Cellular process Developmental process
3 29
5 4 25 Response to stimulus Biological regulation
4
Biogenesis Reproduction
7
7 C
Single organism process Signaling
Biological process
7 (%)
7 Localization Growth
7
23
13
14
24

Figure 13.2  Gene ontology annotations of differentially expressed p ­ roteins

in climacteric (outer-ring) and nonclimacteric (inner-ring) species a­ nalyzed
with Blast 2GO. Overrepresented ripening-related fruit proteins are expressed
in relative percentage of proteins and categorized according to their (A) cel-
lular compartments, (B) molecular functions, and (C) biological processes.

respectively). The high representation for these categories in both climac-

teric and nonclimacteric suggests their importance in the transmission of
genetic information, energy transfer, and storage activities in cells during
fruit ripening. They are involved in a wide range of cellular events, ranging
from activation of signaling cascades via G-proteins to serving as cofactors

424
Proteomics of Fruit Development and Ripening

for enzymes to the gating of plant ion channels via cyclic nucleotides (Talke
et al., 2003; Zelman et al., 2012). Importantly, one group of nucleotide bind-
ing protein, the P-loop-containing cell wall hydrolases, is involved in fruit
softening (Fischer and Bennett, 1991; Thumdee et  al., 2007), probably
explaining their overrepresentation during fruit ripening. The representa-
tion of the RNA binding category was, however, lower in nonclimacteric
species (5%) than in climacteric species (10%). Two molecular categories,
the structural molecular activity and transporter activity, contributed 7%
each and were present only in nonclimacteric species (Figure 13.2B).
In terms of biological process categories, both climacteric and non-
climacteric species showed overrepresentation of proteins involved in the
same 12 categories. The metabolic, cellular, and response to stimulus and
signaling categories were the most represented, contributing more than
60% to the responsive proteins (Figure 13.2C). No significant differences
of category ratios between climacteric and nonclimacteric species were
observed. In accordance with Molassiotis et  al. (2013), the metabolism
category was the most represented in both climacteric and nonclimacteric
species. The authors also reported a large number of common proteins,
suggesting that similar regulatory mechanisms occur in climacteric and
nonclimacteric species (Molassiotis et al., 2013). Of interest is that meta-
bolic processes play a major role during fruit development, which is char-
acterized by massive cellular and physiological changes, including cell
division and expansion, which slows down with maturity and ripening onset
(Figure 13.3).

S MD NTR RIPE

Cell division Ripening

Peak of cell expansion
Cell expansion

Figure 13.3  General fruit cellular and physiological changes during

development and ripening. S, early fruit development stages; MD, mid-
fruit development characterized by extensive cell division and expansion;
NTR, end of development and beginning of fruit ripening; RIPE, ripening
of the fruit. (Adapted from Marondedze, C. et al., Horticulture Research 1,
14039, 2014.)

425
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Overall, this global GO enrichment analysis revealed that overrep-

resented differentially expressed proteins from both climacteric and non-
climacteric species display common biological processes and molecular
functions and are represented across very similar cellular compartments
and organelles. These overrepresented proteins suggest that fruit ripening
encompasses highly active cellular metabolic activities that occur predomi-
nantly in the plastid, cytosol, mitochondria, and nucleus, as well as across
membranes. Further investigations of the pathways in which these overrep-
resented responsive proteins may be involved could possibly give insights
into the mechanisms involved in fruit ripening in both the climacteric and
nonclimacteric fruit species.

13.5  Pathway Analysis Using KEGG

13.5.1  Proteins Associated with Carbohydrate Metabolism

Fruit development and ripening involves variation of organic acid concen-
trations, such as malate and citrate, in the vacuoles of mesocarp cells; these
acids are known to contribute to fruit organoleptic properties (Rocco et al.,
2006). The release and breakdown of these organic acids at the onset of
ripening are critical to fuel the respiratory climax, a key process facilitat-
ing ripening (Rocco et al., 2006). In comparative fruit proteomics studies,
­primary metabolic processes represent the most dominant category in
terms of number of responsive proteins during fruit development and rip-
ening in both climacteric and nonclimacteric species (Guarino et al., 2007;
Prinsi et  al., 2011; Nogueira et  al., 2012; Bianco et  al., 2013; D’Ambrosio
et al., 2013; Marondedze et al., 2014; H.X. Wu et al., 2014). These primary
­metabolic processes include the synthesis of starch, sucrose, fructose, man-
nose, and galactose. For the starch and sucrose metabolism pathways, 12
and 19 enzymes were identified as differentially regulated during develop-
ment and ripening in climacteric and nonclimacteric species, respectively;
of these, 9 were common to the two ripening categories (Figure 13.4). Here,
distinct proteins between climacteric and nonclimacteric species were
identified. In climacteric species only, responsive proteins included pectin
depolymerase (EC3.2.1.15), adenylyltransferase (EC2.7.7.27), and glycoge-
nase (EC3.2.1.1), while in the nonclimacteric species, the expression of
enzymes such as 1,4-β-xylosidase (EC3.2.1.37), glucokinase (EC2.7.1.2),
glucose-6-phosphate isomerase  ( EC5.3.1.9), phosphoglucomutase (EC5.
4.2.2),  4-α-glucanotransferase  (EC2.4.1.25), α-glucosidase (EC3.2.1.20),
UDP-glucose-6-dehydrogenase (EC1.1.1.22), sucrose phosphate phos-
phatase (EC 3.1.3.24), and sucrose phosphate synthase (EC2.4.1.14) was
detected (Figure 13.5A).

426
 Glycolysis/Gluconeogenesis Pyruvate metabolism TCA cycle
c nc c nc c nc

6 5 8 5 11 2

CO2
Starch and sucrose Phosphoenol- coA
metabolism Glucose Glu-6P 2 x Pyruvate Acetyl-coA
pyruvate Citrate
c nc NAD+ NADH Ribulose-1,5P

c nc Carbon fixation
4 8 10 CO2 c nc
Galactose Gly 3-P + Fru-6P Oxaloacetate Glycerate-3P
metabolism 2 6
c nc 2 16 2
H2O
1 4 5 Fructose/mannose PPP Amino acid metabolism
Amino and nuclotide c nc
metabolism
sugar metabolism c nc
7 13 9
1 7 4 Glyoxylate and dicarboxylate metabolism Galactose metabolism

His, Arg, Pro Ala, Asp and Glu Cys and Met Val, Leu and Isoleu Phenylalanine Gly, Ser and Thr

Phenylpropanoid
biosynthesis
Ethylene biosynthesis
4-Coumaroyl-CoA
Nucleotide metabolism Flavonoid biosynthesis
c nc c nc

8 9 3 4 3
Anthocyanidins and anthocyanins

Figure 13.4  Fruit development and ripening-related metabolic pathways.

427
Proteomics of Fruit Development and Ripening
 STARCH AND SUCROSE METABOLISM
UDP-D-
Amino sugar and galacturonate Pectin Pectate 3.2.1.15 D-galacturonate
2.4.1.43 3.1.1.11
nucleotide sugar metabolism 3.2.1.67

428
5.1.3.6
D-Xylose 1,4β-D-Xylan β-D-Glucuroroside
Pentose and glucuronate
32.1.37 2.4.2.24 4.1.1.35 2.4.1.17 3.2.1.31 Ascorbate metabolism
interconversions
UDP-D-xylose UDP-D- D-Glucuronate
glucuronate
3.2.1.28
Sucrose-6P Retinol Trehalose D-Glucose
Sucrose
2.7.1.69 metabolism 24.1.245 2.4.1.64
(extracellular) Sucrose-6P
(2,6-β-D- 1.1.1.22 βD-Glucose-6P
2.4.1.14
Fructosyl)m+n (2,6-β-D-Fructosyl)n 5.4.2.6
3.2.1.26 3.2.1.65 2.4.1.10 3.1.3.12 βD-Glucose-1P
3.2.1.64 3.1.3.24 32.1.122 D-Glucose-6P
1.199.13
α-D-Glucose-6P 2.4.1.15
3.2.1.20 3-Ketosucrose Trehalose-6P 32.1.93
P OST H AR V EST

β-D-Fructose 3.2.1.26 2.4.1.13 2.4.1.11 32.1.122 2.4.18

Sucrose UDP-glucose 2.7.1.69
3.2.1.48 TreXYZ
2.4.1.4 2.4.1.5
2.4.1.10 2.7.7.9 Maltose-6’P 5.499.16
D-Fructose α,α-Trehalose
1,3-β-Glucan D-Fructose (extracellular)
D-Glucose 3.2.1.39 2.4.1.34 2.7.1.69
Cyclomahodextrin
3.2.1.58 2.4.1.7 Sucrose 3.1.3.90
Maltose
3.2.1.21 2.4.1.35 2.7.7.27 2.4.1.21
3.2.1.54 (extracellular)
α-D-Glucose Glucoside
Mahodextrin
32.1.74
2.4.1.12 ADP-glucose Starch;
2.7.1.1 2.7.1.4 Cellobiose 3.6.1.9 3.6.1.21 Glycogen
GDP-glucose Maltose
3.2.1.21 3.2.1.4 2.4.1.1 3.2.1.2
2.4.1.29 2.7.7.34
RI P ENING

Dextrin
3.2.1.21 3.2.1.91 Cellulose
α-D-Glucose- 3.2.1.1
β-D-Glucose 1,4-β-D-Glucan 1P Glycogen; 2.4.1.25 3.2.1.20
3.6.1.21
Amylose 3.2.1.3 3.2.1.10 2.7.1.175
Amino sugar and
nucleotide sugar metabolism 2.7.7.33 2.4.1.18 3.2.1.3
CDP-glucose α-D-Glucose
3.2.1.33
2.7.1.41 2499.16
α-Maltose-1-P
2.7.1.10
Glycolysis/ 3.2.1.10
α-D-Glucose-1,6P2 2.7.1.106 5.4.2.2 Gluconegenesis 2.4.1.20
Isomaltose
3.1.3.9 α-D-Glucose-6P
Cellohexaose Cellotetraose
2.7.1.1 3.2.1.74 3.2.1.74 3.2.1.74 3.2.1.74 3.2.1.74
Celloheptaose Cellopentaose Cellotriose Cellobiose
α-D-Glucose 2.7.1.2 5.3.19
P H Y SIOLOG Y

β-D-Fructose-6P
(A)

Figure 13.5A  Overview of the (A) starch and sucrose metabolisms, (B) fructose and mannose metabolisms, and (C)
­galactose metabolism. These are some of the most represented metabolic pathways during fruit development and rip-
ening. The boxes highlight proteins with differential accumulation during fruit development and ripening. The symbols
highlight proteins whose expression significantly varies in the categories represented.
 FRUCTOSE AND MANNOSE METABOLISM

L-Sorbose
D-Allose
1.19921
α-D-Glucose 1.1.2.2 2.7.1.55
Galactose
1.1.1.21 D-Sorbitol
metabolism 1.1.1.11 D-Mannitol-1P
1.1.1.67 3.1.3.22 D-Allose-6P
5.3.1.5 1.1.1.14 1.1.1.15 2.7.1.69
1.1.1.138 D-Mannitol
RpiB
D-Fructose-2,6P2
3.1.3.- 3.1.3.54
D-Fructose D-Fructose-2P 1.1.1.17 D-Allulose-6P
3.2.1.80 Fructan 3.1.3.46
5.3.1.7 2.7.1.1 2.7.1.4 2.7.1.105 5.1.3.-
2.7.1.69 β-D-Fructose-6P
Amino sugar and D-Mannose D-Mannose-6P
nucleotide sugar 2.7.1.1 5.3.1.8 Glycolysis
metabolism Amino sugar and
2.7.1.7
5.4.2.8 nucleotide sugar metabolism 1.1.1.140
ADP-Mannose D-Mannose-1P
3.6.1.21 GDP- D-Sorbitol-6P
3.2.1.78 3.2.1.77 3.2.1.137 D-mannuronate 4.2.2.3 Dinner
2.4.1.33
4.2.2.11 3.1.3.11 2.7.1.11 1.1.1.-
2.4.1.- 2.7.7.13 2.7.7.22 3.6.1.21 Alginate
Mannan 1.1.1.132 Mannosyl-
2.7.1.90 2.7.1.69
3-phosphoglycerate L-Sorbose-1P
2.4.1.- 2.41.217 3.1.3.70 Mannosyl-
1,4-β-Mannan GDP-D-mannose glycerate
D-Sorbitol 2.7.1.69
2.41.269
4.2.1.47
N-Glycan GDP-4-oxo-6-deoxy-
biosynthesis L-Sorbose
D-mannose
1.1.1.135
β-D-Fructose-1,6P2
GDP-6-deoxy-D-talcose
1.1.1.271
4.12.13
1.1.1.187
GDP-L-fucose
1.1.1.281 Glyceraldehycle-3P
GDP-D-rhamnose Glycolysis
2.7.7.30
L-Fucose
2.7.1.52 5.3.1.1
L-Fucose-1P
5.3.1.25
L-Fuculose-1P Glycerone-P
2.7.1.51 4.1.2.17 2.7.1.28 2.7.1.56
L-Fuculose
L-Lactate
Pyruvate
1.1.1.122 3.1.1.- 4.2.1.68 1.1.1.- 3.7.1.-
metabolism
L-Fucono- L-Fuconate 2-Dehydro-3-deoxy- 2,4-Diketo-3-deoxy-
lactone L-fuconate L-fuconate
L-Rhamno- D-Glycer-
furanose L-Rhamnonate
aldehyde
1.1.1.173 3.1.1.65 4.2.1.90 4.1.2.53
L-Rhamnono- L-Lactaldehyde
2-Dehydro-3-deoxy-
1,4-lactone L-rhamnonate
L-Rhamnose

5.3.1.14
L-Rhamnulose-1P
2.7.1.5 4.1.2.19
L-Rhamnulose
2.7.1.3
4.1.2.13
2.7.1.69 D-Fructose-1P
(B)

Figure 13.5B  (Continued )

429
Proteomics of Fruit Development and Ripening
430
GALACTOSE METABOLISM

1.1.3.9
2-Dehydro-3-deoxy-
1.1.1.48 4.2.1.6 D-galactonate 4.1.2.55 D-Glyceraldehyde
3.1.1.25
1.1.1.120 D-Galactonate 4.2.1.140 4.1.2.51
D-Galctomono-1,4-lactone
1.1.1.360
2.7.1.58 2.7.1.178
1.1.1.359 D-Galactono-1,5-lactone
UDP-glucose 4.1.2.21 D-Glyceraldehyde-3P
Pentose and glucuronate Pentose phosphate
Amino sugar and nucleotide
interconversions 4.1.2.55 pathway
sugar metabolism 2-Dehydro-3-deoxy-
P OST H AR V EST

2.7.7.9 D-galactonate-6P
D-Galactose α-D-Galactose-1P
α-D-Glucose-1P
5.1.3.3 2.7.1.6 2.7.7.12 5.1.3.2 Glycerone-P
α-D-Galactose
2.7.7.10 2.7.7.64
4.1.2.40
UDP-galactose Galactinol
3.2.1.23 2.41.123
Galactan
5.4.99.9
3.2.1.23 Lactose 2.4.1.82
UDP-D- 5.4.2.2
2.4.1.22 Raffinose
3.2.1.108 galactofuranose
3.2.1.22 Sucrose
3.1.3.9 N-Acetyl-
RI P ENING

2.7.1.69 2.4.1.67 D-galactosamine

2.7.1.1 D-Tagatose-1,6P2

α-D-Glucose 3.2.1.22 Stachyose

2.7.1.1.2 α-D-Glucose-6P
3.2.1.26
3.2.1.22 Galactinol 3.2.1.26 2.7.1.69
3.2.1.20 3.2.1.10
Lactose-6’P Glycolysis
D-myo-Inositol Manninotriose D-Galactosamine N-Acetyl-
D-galactosamine-6P
3.2.1.22 Melibiitol Fructose and mannose 3.2.1.22
metabolism Mehbiose 2.7.1.11 2.7.1.144
D-Sorbitol 3.5.1.25
3.2.1.85 3.2.1.22 2.7.1.69
3.2.1.22 Epimelibiose
5.3.1.-
D-Mannose D-Galactosamine-6P
D-Galactose D-Glucose D-Fructose
Galactosyl-
P H Y SIOLOG Y

3.2.1.22
glycerol 5.3.1.26
D-Galactose-6P
Glycerol
Galactitol D-Tagatose
1.1.1.21 1.1.1.16 2.7.1.101 D-Tagatose-6P
(C)
2.7.1.69 1.1.1.251
Galactitol-1P

Figure 13.5C  (Continued )

Proteomics of Fruit Development and Ripening

For the fructose and mannose metabolisms, 8 and 11 differentially

regulated proteins were identified from climacteric and nonclimacteric
species, respectively, and of these, 7 were common. Of the seven common
proteins, five play a role in the glycolysis pathway as well as in fructose
metabolism (Figure 13.4). Most of the responsive proteins associated
with  mannose metabolism were identified in nonclimacteric species.
Among these, mannose-6-phosphate isomerase (EC5.3.1.8), phosphoman-
nomutase (EC5.4.2.8), dolichol-phosphate-mannose, GDP-mannose-4,6-
dehydratase (EC4.2.1.47), and L-lactate dehydrogenase (EC1.1.1.27) were
identified (Figure 13.5B).
Five and nine responsive proteins from the galactose metabolism
were identified in climacteric and nonclimacteric fruit species, respectively
(Figure 13.4), of which four were common. Interestingly, two enzymes,
invertase or acid β-fructofuranosidase (EC3.2.1.26) and glycoside hydro-
lase or melibiase (EC3.2.1.22), which are involved in the biosynthesis of
D-galactose from raffinose, were present in both climacteric and noncli-
macteric species. Only galactosyltransferase (EC2.4.1.67) was unique to
nonclimacteric fruits (Figure 13.5C) and is involved in the stachyose biosyn-
thesis. Lactase, also called β-galactosidase (EC3.2.1.23), was identified as
differentially regulated in nonclimacteric species only (Figure 13.5C). This
enzyme is involved in the hydrolysis of lactose and galactan to D-galactose.
Overall, a carbohydrate metabolism program is crucial to the regulation
of biosynthesis of simple sugars that are needed for energy provision during
fruit ripening. The identification of a greater number of proteins related to
carbohydrate metabolism in nonclimacteric fruit species suggests that dif-
ferent processes for sugar metabolism and accumulation might be involved
in these fruit species in comparison to their climacteric counterparts.

13.5.2  Proteins Associated with Energy Metabolism

13.5.2.1  Proteins with a Role in Carbon Fixation

Out of the 25 enzymes involved in the carbon fixation pathway, 22 have
been identified as responsive to fruit development and ripening. Of the
latter, 16 were common to climacteric and nonclimacteric species. This
large proportion of responsive proteins compared to the total number of
proteins identified from this pathway is an indication that the fruit devel-
opment and ripening process is a high-energy metabolically active event.
Responsive proteins unique to the climacteric species include alanine
transaminase (EC2.6.1.2), which catalyzes the reversible transamina-
tion reaction involving the transfer of an amino group from L-alanine to
α-ketoglutarate to give pyruvate and L-glutamate; ribulose-phosphate-
3-epimerase (EC5.1.3.1); and transketolase (EC2.2.1.1), which catalyzes

431
P OST H AR V EST RI P ENING P H Y SIOLOG Y

the biosynthesis of ribulose-5­-phosphate from xylulose-5-phosphate and

ribose-5-phosphate. Transketolase and transaldolase are enzymes that
link the glycolytic and pentose phosphate pathways. On the other hand,
phosphoenolpyruvate carboxylase (EC4.1.1.31) and an NADP-dependent
malate dehydrogenase (EC1.1.1.82) were the two enzymes unique to non-
climacteric species. Phosphoenolpyruvate carboxylase plays an antipleu-
rotic role in plant cells, supplying oxaloacetate to the TCA cycle to replenish
the intermediates removed for amino acid biosynthesis (Eikmanns et al.,
1989). Malate dehydrogenase catalyzes oxaloacetate into malate, which is
a critical step that allows evasion of the effects of photorespiration.

13.5.2.2  Proteins Associated with the Pentose Phosphate,

Glycolysis, and Pyruvate Metabolisms
All the primary enzymes involved in the pentose phosphate pathway
(PPP) were identified as differentially regulated during fruit maturation
(Figure 13.4). In climacteric fruit species, two enzymes involved in the metab-
olism of ribulose-5-phosphate, ribulose-phosphate-3-epimerase (EC5.1.3.1),
and ribulose-5-phosphate isomerase (EC5.3.1.6) were detected as respon-
sive (Figure 13.6A), while in nonclimacteric species, glucose-6-phosphate
isomerase (EC5.3.1.9), which catalyzes the reversible enzymatic conversion
of glucose-6-phosphate to fructose-6-phosphate, was identified.
Glycolysis is an important metabolic process that converts glucose
to pyruvate while generating ATP (energy) and NADH (reducing power).
It is an essential pathway that produces key precursor metabolites such
as glucose-6-phosphate and fructose-6-phosphate, as well as phospho-
enolpyruvate. The final product, pyruvate, undergoes oxidative decarbox-
ylation to form acetyl-CoA, an important metabolite precursor that links
glycolysis to the tricarboxylic acid (TCA) cycle or citric acid cycle, which
are the two major metabolic pathways that generate energy. In the glycol-
ysis and pyruvate metabolism pathways, other responsive enzymes were
identified during fruit development and ripening in the nonclimacteric
species (Figure  13.4). Of interest, accumulation of a number of proteins
involved in the early glycolytic program varied with fruit development of
nonclimacteric species. These included glucokinase (EC2.7.1.2), which
­phosphorylates glucose using ATP to give rise to glucose-6-phosphate and
ADP; glucose-6-phosphate isomerase (EC5.3.1.9), which catalyzes the
reversible conversion of glucose-6-phosphate to fructose-6-phosphate; and
glyceraldehyde-3-phosphate dehydrogenase (NADP+) (EC1.2.1.9), which
induces the reversible conversion of 1,3-diphosphoglycerate to glyceralde-
hyde-3-phosphate, a central step in glycolysis and gluconeogenesis. The
rest of the responsive proteins were detected in both climacteric and non-
climacteric species (Figure 13.6B).

432
 Glycolysis Gluconeogenesis

ATP
Glucose Pi
Glucose-6-phosphate (P) hexokinase Glucose-6-phosphatase
ADP H2O
NADP+ Glucose-6-P Glucose-6-
* dehydrogenase phosphate (P)
NADPH
6-phosphogluconolactone
H2O
* Gluconolactonase ATP Fructose-6-P H2O
H+ phospho-
H2O * * Fructose 1,6-bisphosphatase
fructokinase-1
6-phosphogluconate ADP

Oxidative phase
Fructose H2O
NADP+
* 6-phosphogluconate 1,6-bisP
* NADPH + CO2 dehydrogenae
*
Dihydroxyacetone P Dihydroxyacetone P
Ribulose-5-P
Ribulose-5-P isomerase Ribulose-5-P 3-epimerase
*
Ribulose-5-P Xylulose-5-P 2x Glyceraldehyde 3-P
Transketolase 2 (NAD++Pi) 2 (NAD++Pi)
* *
2 (NADH+H+) 2 (NADH+H+)
Glyceradehyde-3-P + Sedoheptulose-7-P

Non-oxidative phase
2x 1,3-bisphosphoglycerate
* Transaldolase 2x ADP 2x ADP
*
Fructose-6-P + Erythrose-4-P Xylulose-5-P 2x ATP 2x ATP
Transketolase 2x 3-phosphoglycerate
*
Glycolysis
Glyceraldehyde-3-P + Fructose-6-P 2x phosphoenol- 2x GDP
Climacteric only pyruvate PEP carboxykinase
non-Climacteric only 2x ADP 2x GTP
* both Climacteric and non-Climacteric Glycolysis * 2x oxaloacetate
fruit categories 2x ATP
2x ADP
(A) (B) 2x pyruvate pyruvate carboxylase
2x ATP

Figure 13.6  Overview of the (A) pentose phosphate pathway and (B) glycolysis and gluconeogenesis pathways. Common
and climacteric- and nonclimacteric-specific enzymes’ responsive proteins during fruit development and ripening are shown.

433
Proteomics of Fruit Development and Ripening

Symbols highlight proteins whose expression significantly varied.

P OST H AR V EST RI P ENING P H Y SIOLOG Y

The expression of proteins related to the pyruvate metabolism, an

essential pathway that links glycolysis and TCA cycles for the biosynthe-
sis of acetyl-CoA, was also responsive to fruit development and ripening.
Variations in protein regulation between climacteric and nonclimacteric
fruits were observed, with the exception of pyruvate phosphate dikinase
(EC2.7.9.1), an enzyme that catalyzes the reversible conversion of pyru-
vate to phosphoenolpyruvate and pyruvate kinase (EC2.7.1.40), which is
involved in the conversion of phosphoenolpyruvate to pyruvate with con-
comitant phosphorylation of ADP to ATP (Figure 13.7A). Overall, pyruvate

1.2.1.22 Methylglyoxal L-Lactaldehyde

1.2.1.23 1.1.1.283
1.2.1.49
Glycolysis Glycine, serine and
threonine metabolism 1.1.1.78 D-Lactaldehyde
4.1.1.31
4.1.1.78 Acetylene- 1.1.1.79
Phosphoenol- dicarboxylate
4.1.1.32 pyruvate (R)-S-Lactoyl- 1.2.1.23 1.2.1.22
4.1.1.78 glutathione
4.1.1.38
4.4.1.5 3.1.2.6
4.1.1.49 2.7.9.1 2.7.9.2 4.1.1.-
2-Hydroxyethylene-
42.1.130
dicarboxylate
Nicotinate and nicotinamide
1.1.1.28 D-Lactate
metabolism 2.7.1.40
1.1.2.4
Valine, leucine and Pyruvate 1.1.2.5 5.1.2.1
isoleucine biosynthesis 1.1.2.3
1.1.1.27 L-Lactate

1.1.99.7
1.13.124
4.1.1.3 1.2.4.1 1.2.5.1
2-Hydroxy- 1.2.3.3
6.4.1.1 ThPP Acetyl-P
ethyl-ThPP 2.7.2.12
1.1.1.38 1.1.1.39
2.7.2.1
1.1.1.40 Lipoamide-E 1.2.4.1 1.2.7.1 1.2.3.6

2.3.1.54 S-Acetyl-
Formate
Oxaloacetate 1.8.1.4 dihydro- 3.6.1.7 4.1.2.36
lipoamide-E 2.3.1.8 EutD
Dihydro-
lipoamide-E 2.3.1.12

Glyoxylate
1.1.1.37 1.1.1.82
metabolism 3.1.2.1 Acetate
1.1.5.4 Acetyl-CoA
2.8.3.18
Propanoate 2.8.3.1
Glyoxylate 6.2.1.13
metabolism metabolism
6.2.1.1 6.2.1.1
(S)-Malate Acetyladenylate 1.2.1.3 1.2.1.-
4.2.1.2 1.3.5.4
Fumarate Succimate 1.2.99.3 1.2.99.6

2.3.3.9 1.2.1.10
Citrate cycle Acetaldehyde
2.3.3.13 Leucine biosynthesis
(R)-2-Ethylmate 2.3.3.6 3-Carboxy-3-hydroxy-
Acetoacetyl-CoA 4-methylpentanoate
Butanoate metabolism 2.3.1.9 2.3.3.14 Lysine biosynthesis
Homocitrate
Synthesis and degradation
of ketone bodies 4.1.3.- 6.4.1.2 Fatty acid biosynthesis
2-Propylmalate Malonyl-CoA

(A) - climacteric only

- non-climacteric only

Figure 13.7A  Enzymes involved in the pyruvate pathway and tricarboxylic

acid cycle. The pathways of (A) pyruvate metabolism and the (B) tricarbox-
ylic acid cycle show the differentially regulated proteins during fruit devel-
opment and ripening. The colored boxes highlight proteins with differential
accumulation during fruit development and ripening. The symbols highlight
proteins whose expression significantly varied in the categories represented.
(Adapted from KEGG using Blast2GO.)

434
Proteomics of Fruit Development and Ripening

kinase, together with phosphofructokinase and hexokinase, regulates

glycolysis and thus impacts energy generation during fruit growth and
ripening. Pyruvate dehydrogenase (EC1.2.4.1), malate oxidoreductase
(EC1.1.1.38), decarboxylating malate dehydrogenase (EC1.1.1.39), and
oxaloacetate-decarboxylating malate-dehydrogenase NADP+ (EC1.1.1.40),
which catalyzes the oxidative decarboxylation of malate to form pyruvate, a
reaction important for generating the CO2 used in sugar production d­ uring
the Calvin cycle, were also identified. Expression of phosphoenolpyru-
vate carboxylase (EC4.1.1.31), NADP-dependent malate dehydrogenase
(EC1.1.1.82), phosphoenolpyruvate carboxykinase (EC4.1.1.49), oxaloac-
etate decarboxylase β-subunit (EC4.1.1.3), and pyruvate dehydrogenase
E2 component (EC2.3.1.12) was detected as differentially regulated, but
in nonclimacteric species only (Figure 13.7A). These five proteins are also
involved in regulating the fate of pyruvate and phosphoenolpyruvate.

Phosphoenol-
41.1.32 pyruvate Glycolysis/
41.1.49 Gluconeogenesis
Fatty acid biosynthesis

Fatty acid eloggation in

1.2.7.1
mitochondria ThPP

Val, Leu & Ile degradation 2-Hydroxy-

ethyl-ThPP
Fatty acid metabolism 231.1.12 1.2.4.1 1.2.4.1 Pyruvate
Acetyle-CoA S-Acetyldihydro-
Alamine, asperteme and lipoamide-E
glucamate metabolism 1.8.1.4 6.4.1.1
Dihydro- Lipoamide-E
Glycorydate and lipoamide-E
dicarboxylate metabolism
233.1 Citrate Isocitrate
1.1.1.37 4.2.1.3 4.2.1.3
Orbacetate 233.8
cis-Aconitate

1.1.5.4
(S)-Malate 1.1.1.42

Tyrosine 42.12
metabolism Oxalosuccinate 1.1.1.41 1.11286
Arginine and
proline metabolism Fumarate
1.1.1.42

135.4 135.1
ThPP 2-Oxo
6.2.1.4 Succinyl-CoA glutarate
6.2.1.5 23.1.61 1.2.42 1.2.42
Succinate 2.8.3.18 S-Succinyl- 3-Carboxyl-
dihydrolipomide-E hydroxypropyl-Thpp Ascorbate and aldarate
metabolism
1.8.1.4
Val, Leu & Ile Dihydro- Lipomide-E Alanine, aspartate and
degradation lipomide-E glutamate metabolism
1.2.7.3
D-Gln & D-Glu
metabolism

Figure 13.7B  (Continued )

435
P OST H AR V EST RI P ENING P H Y SIOLOG Y

13.5.2.3  Proteins Involved in the TCA Cycle

The TCA cycle, the second stage in cellular respiration, is key for the catab-
olism of organic molecules in the presence of oxygen to harness the energy
essential for living cells to grow and divide. The expression of 11 and 13 pro-
teins was identified as responsive to fruit development and ripening in cli-
macteric and nonclimacteric fruit species, respectively, and of these, 11 are
common to both (Figure 13.4). Two of these 11 proteins were detected in
nonclimacteric species: ATP-citrate synthase (EC2.3.3.8) and 2-oxogluta-
rate dehydrogenase (EC1.2.4.2) (Figure 13.7B). The ATP-citrate synthase,
also known as ATP-citrate lyase, catalyzes the conversion of citrate and
coenzyme A to oxaloacetate and acetyl-CoA. The enzyme 2-oxoglutarate
dehydrogenase is a determinant of metabolic fluxes through the TCA cycle
and converts 2-oxoglutarate, coenzyme A, and NAD(+) to succinyl-CoA,
NADH, and carbon dioxide (Tretter and Adam-Vizi, 2005). Overall, the
detection of changes in the expression of TCA cycle–associated proteins
in response to fruit development and the ripening processes suggests that
high metabolic activities regulate energy generation for these processes
(Martinez-Esteso et al., 2011).

13.5.3  Proteins Involved in Amino Acid

Metabolism and Ethylene Biosynthesis
Disparities in the expression of amino acid metabolism-related proteins dur-
ing fruit development and ripening were also noted between climacteric and
nonclimacteric species (Figure 13.4). Notably, variations in the accumula-
tion of proteins involved in the biosynthesis of L-alanine were observed in
climacteric species only, while proteins involved in L-aspartate and succinate
biosynthesis were differentially expressed only in the nonclimacteric spe-
cies. Cysteine and methionine metabolisms are well-known distinguishing
pathways between climacteric and nonclimacteric fruit ripening, mainly
because ethylene biosynthesis necessitates these two amino acids (Bapat
et  al., 2010). As described earlier, climacteric fruit species are character-
ized by a concomitant burst in ethylene synthesis and a peak in respiration
at the onset of ripening, which is not observed in nonclimacteric species
(Palma et al., 2011). Expression of ethylene biosynthesis-related proteins was
responsive to the fruit development and ripening processes in both species
categories, but it mainly increased in climacteric species (Zhang et al., 2008;
Marondedze and Thomas, 2012a; Nogueira et  al., 2012; D’Ambrosio et  al.,
2013; Shi et al., 2014). On the contrary, in nonclimacteric species, their abun-
dance decreased (Giribaldi et al., 2007; Bianco et al., 2009; Martinez-Esteso
et al., 2011). However, expression of 1-aminocyclopropane-1-carboxylic acid
oxidase (ACO), for example, involved in ethylene biosynthesis, had been

436
Proteomics of Fruit Development and Ripening

shown to increase during the véraison in grapes (Chervin et al., 2004; Sun
et al., 2010), at the onset of ripening in strawberries (Trainotti et al., 2005),
and in some cases, during ripening, such as in pineapple (Cazzonelli et al.,
1998), which are all nonclimacteric fruits. Even though a slight elevation in
ethylene has been reported in grape, strawberry, and capsicum ripening as a
result of the increase in ACO expression (Chervin et al., 2004; Iannetta et al.,
2006; Pham, 2007), the level of ethylene was very low compared to that in
climacteric fruits (Aizat et al., 2013). This low level of ethylene in nonclimac-
teric fruits might be compensated, at least partly, by the presence of other
hormones, such as brassinosteroids and abscisic acid, to facilitate ripening in
some fruits, as suggested in grape (Symons et al., 2006) and strawberry (Jia
et al., 2011), but further research is necessary.

13.5.4  Proteins Involved in Flavonoid Biosynthesis

Flavonoids, pigments that give color to flowers, fruits, and seeds, are one of
the widely distributed and multifunctional secondary metabolites in plants.
The biosynthesis of flavonoid involves a series of enzymatic modifications
of 4-coumaroyl-CoA, a product of phenylalanine from the phenylpropanoid
pathway, to yield anthocyanins and proanthocyanidins (tannins), among
other various polyphenols (Teixeira et al., 2013). In plants, flavonoids are
implicated in various biological functions, such as defense against pathogen
infection, UV photoprotection and signaling (Smillie and Hetherington,
1999; Merzlyak and Chivkunova, 2000; Ferreyra et al., 2012), and environ-
mental stress adaptation (Williams et al., 2004; Nascimento and Fett-Neto,
2010). Four enzymes of this pathway, detected in both climacteric and noncli-
macteric species, were noted as differentially expressed during fruit devel-
opment and ripening: chalcone synthase (EC2.3.1.74), chalcone isomerase
(EC5.5.1.6), flavanone-3-dioxygenase (EC1.14.11.19), and leucocyanidin
oxygenase (EC1.14.11.19) (Figures 13.4 and 13.8). Another two responsive
enzymes, flavonol synthase (EC1.14.11.23) and bifunctional dihydroflavo-
nol-4-reductase/flavanone-4-reductase (EC1.1.1.219/EC1.1.1.234), were
identified only in nonclimacteric fruit species. Expression of anthocyanin
biosynthesis-related genes was shown to be positively correlated with
anthocyanin concentrations in ‘Orin’, ‘Fuji’, and ‘Jonathan’ apple cultivars
during ripening (Honda et al., 2002). Anthocyanins have been correlated
with slowing down overripening in some fruits such as tomato (Zhang
et al., 2013). An increase in ethylene has been shown to negatively regulate
anthocyanin synthesis, while an increase in anthocyanins inhibits photo-
synthesis (Das et al., 2011). Thus, a general increase in flavonoid biosynthe-
sis enzymes in nonclimacteric fruits may partly explain the downregulation
of ethylene biosynthesis proteins during the onset of ripening, as shown
previously in dates (Marondedze et al., 2014).

437
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Cinnamoyl-CoA Pinocembrin chalcone Pinocembrin Pinobanksin

Phenylpropanoid 23174 55.1.6 114119 Pinobanksin 3-acetate
biosynthesis
1141122 1141133
Pinostrobin
1141311
Chrysin Galangin
111219
23.1.170 55.1.6 114119 5-Deoxyleucopelargonidin
Isoliquiritigenin Liquiritigenin Garbansol
Isoflavonoid biosynthesis 1141321
1141321
7.4-Dihydroxy-
flavone 111219
55.1.6 114119 5-Deoxyleucocyanidin
Butein Butin Dilydrofisetin

Chalcone 2'-O-glucoside Prunin

Isoflavonoidbiosynthesis 241185 241236 Naringin
Hesperetin
2.4.1.- Desmethylxanthohumol Hesperetin 7-O-glucoside
2.1.1.- 241185 241236 Neohesperidin
Xanthohumol
Naringenin chalcone Naringenin Dihydrokaempferol 114219 Leucopelargonidin (-)-Epiafzelechin
23174 55.1.6 114119 114119 1.3.1.77
114119
p-Coumaroyl-CoA 1.17.13 Pelargonidin
Naringenin chalcone
231133 231133 11.1234 Apiforol
4-O-glucoside
Aureusidin (+)-Afzelechin
1 21 36 Kaempferol
6-O-glucoside
p-Coumaroyl p-Coumaroyl 8-C-Glucosyberigenin Vitexin 1141123 Flavone and flavonol biosynthesis
shikimic acid quinic acid
Apigenin 1141388 1141321
1141336 1.1413. 1141336
1 21 36 1141122
1141388 1141321
Caffeoyl Caffeoylquinic acid Flavone and flavonol
shikimic acid 1141388 1141321 biosynthesis
1141123 Quercetin
231133 231133 1 21 36 Bracteatin
6-O-glucoside 1141388 1141321
2',3,4,4',6'-Pentahydroxy
-chalcone 4-O glucoside 111219 Leucocyanidin Cyanidin (-)-Epicatechin
23174 114119 1141119 1.3.1.77
Caffeoyl-CoA 2',3,4,4',6'-Pentahydroxychalcone Eriodictyol Dihydroquercetin 1141119
1.17.1.3
1141122 Anthocyanin biosynthesis
Luteolin
(+)-Catechin
21.1.104
111234 Luteoforol

1141133 1141133 1141388 1141388

23174 1141122 1141123 Myricetin

Feruloyl-CoA 4,2',4',6'-Tetrahydroxy- Homoeriodictyol Tricetin
Leucodelphinidin Delphinidin (-)-Epigallocatechin
3-methoxychalcone
114119 111219 1141119 1.3.1.77
Dihydrotricetin Dihydromyricetin 1.17.1.3

(+)-Gallocatechin

non-climacteric only

Figure 13.8  Enzymes involved in the flavonoid biosynthesis pathway. The

pathway shows enzymes that are differentially regulated and play a role
during fruit development and ripening. The shaded boxes highlight pro-
teins with differential accumulation during fruit development and ripening.
The symbols highlight proteins whose expression significantly varied in the
categories represented. (Adapted from Kanehisa, M., and S. Goto. 2000.
KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Research,
28, 27–30).

13.6  Conclusions and Future Perspectives

Generally, much progress has been made in plant proteomics, and this
technology has proven invaluable for deciphering both physiological
and developmental roles in plants. With the extension of proteomics
approaches to fruit studies, the identification of novel proteins as bio-
markers for different traits, such as those governing quality and yield,
has progressed tremendously. In this work, an overview and reanalysis
of recentreports on fruit development and ripening proteomics is pro-
vided using a representative set of the most studied and documented
fruits in order to obtain a global view of cellular processes involved
in fruit development and ripening. Further, these data were used to

438
Proteomics of Fruit Development and Ripening

characterize the biology underlying climacteric and nonclimacteric

fruit species metabolism and the characteristics that distinguish these
two categories during development and ripening. We note that both cli-
macteric and nonclimacteric species share common developmental and
ripening profiles involving enrichment of biological processes such as
the metabolic process, response to stimuli, biological regulation, and
signaling. In addition, numerous proteins play an enzymatic role in the
metabolism pathways, such as carbohydrate, energy, and amino acid
metabolisms and flavonoid biosynthesis, albeit with a modification or
differential expression during development and ripening. As more pro-
teomics data from a greater representation of economically important
fruits will emerge in the near future, a global analysis such as this is
encouraged to draw parallels and highlight differences that can provide
more useful information for any potential biotechnological strategies and
implementations. Concomitantly, proteomic tools and approaches that
specifically cater to the needs of fruit proteomics must improve, as cur-
rent methods are lagging in terms of proteome coverage, sensitivity, and
reproducibility. For example, in apples and grapes, varying proteomics
data have been reported in several studies adopting different techniques
(Giribaldi et al., 2007; Di Carli et al., 2011; Martinez-Esteso et al., 2011;
Marondedze and Thomas, 2012b; Kambiranda et al., 2014). As the field
continues to mature, proteomics can complement metabolomics, tran-
scriptomics, and genomics to provide a more accurate and wholesome
representation of the cellular processes involved in the development and
ripening of fruits.

References
Adams-Phillips, L., Barry, C., and Giovannoni, J. 2004. Signal transduction
systems regulating fruit ripening. Trends in Plant Science 9, 331–338.
Aizat, W.M., Able, J.A., Stangoulis, J.C.R., and Able, A.J. 2013. Proteomic anal-
ysis during capsicum ripening reveals differential expression of ACC
oxidase isoform 4 and other candidates. Functional Plant Biology 40,
1115–1128.
Alba, R., Payton, P., Fei, Z., McQuinn, R., Debbie, P., Martin, G.B., Tanksley,
S.D., and Giovannoni, J.J. 2005. Transcriptome and selected metabolite
analyses reveal multiple points of ethylene control during tomato fruit
development. Plant Cell 17, 2954–2965.
Andrade, J.D., Toledo, T.T., Nogueira, S.B., Cordenunsi, B.R., Lajolo, F.M.,
and do Nascimento, J.R.O. 2012. 2D-DIGE analysis of mango (Mangifera
indica L.) fruit reveals major proteomic changes associated with ripen-
ing. Journal of Proteomics 75, 3331–3341.

439
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Bapat, V.A., Trivedi, P.K., Ghosh, A., Sane, V.A., Ganapathi, T.R., and Nath, P.
2010. Ripening of fleshy fruit: molecular insight and the role of ethylene.
Biotechnology Advances 28, 94–107.
Bianco, L., Alagna, F., Baldoni, L., Finnie, C., Svensson, B., and Perrotta, G.
2013. Proteome regulation during Olea europaea fruit development.
PLoS One 8, e53563.
Bianco, L., Lopez, L., Scalone, A.G., Carli, M.D., Desiderio, A., Benvenuto,
E., and Perrotta, G. 2009. Strawberry proteome characterization and its
regulation during fruit ripening and in different genotypes. Journal of
Proteomics 72, 586–607.
Bonghi, C., and Manganaris, G.A. 2012. Systems biology approaches reveal
new insights into mechanisms regulating fresh fruit quality. In OMICs
Technologies Tools for Food Science, ed. Benkeblia, N., 201–206. CRC
Press, Boca Raton, FL.
Bouvier, F., and Camara, B. 2006. The role of plastids in ripening fruits. In
The Structure and Function of Plastids, ed. Wise, R.R., and J.K. Hoober,
419–432. Springer, Netherlands.
Camara, B., Hugueney, P., Bouvier, F., Kuntz, M., and Monegar, R. 1995.
Biochemistry and molecular biology of chromoplast development.
International Review of Cytology 163, 175–247.
Carpentier, S.C., Witters, E., Laukens, K., Deckers, P., Swennen, R., and
Panis, B. 2005. Preparation of protein extracts from recalcitrant plant
tissues: An evaluation of different methods for two-dimensional gel elec-
trophoresis analysis. Proteomics 5, 2497–2507.
Cazzonelli, C.I., Cavallaro, A.S., and Botella, J.R. 1998. Cloning and charac-
terisation of ripening-induced ethylene biosynthetic genes from non-­
climacteric pineapple (Ananas comosus) fruits. Australian Journal of
Plant Physiology 25, 513–518.
Chaves, A.L.S., and de Mello-Farias, P.C. 2006. Ethylene and fruit ripening:
from illumination gas to the control of gene expression, more than a
century of discoveries. Genetics and Molecular Biology 29, 508–515.
Chervin, C., El-Kereamy, A., Roustan, J.P., Latche, A., Lamon, J., and
Bouzayen, M. 2004. Ethylene seems required for the berry develop-
ment and ripening in grape, a non-climacteric fruit. Plant Science 167,
1301–1305.
Ciardi, J.A., Tieman, D.M., Lund, S.T., Jones, J.B., Stall, R.E., and Klee, H.J.
2000. Response to Xanthomonas campestris pv. vesicatoria in tomato
involves regulation of ethylene receptor gene expression. Plant
Physiology 123, 81–92.
Conesa, A., and Gotz, S. 2008. Blast2GO: A comprehensive suite for functional
analysis in plant genomics. International Journal of Plant Genomics
2008, 619832.
Conesa, A., Gotz, S., Garcia-Gomez, J.M., Terol, J., Talon, M., and Robles,
M. 2005. Blast2GO: A universal tool for annotation, visualization and
­analysis in functional genomics research. Bioinformatics 21, 3674–3676.

440
Proteomics of Fruit Development and Ripening

D’Ambrosio, C., Arena, S., Rocco, M., Verrillo, F., Novi, G., Viscosi, V., Marra,
M., and Scaloni, A. 2013. Proteomic analysis of apricot fruit during rip-
ening. Journal of Proteomics 78, 39–57.
Dandekar, A.M., Teo, G., Defilippi, B.G., Uratsu, S.L., Passey, A.J., Kader,
A.A., Stow, J.R., Colgan, R.J., and James, D.J. 2004. Effect of down-­
regulation of ethylene biosynthesis on fruit flavor complex in apple
fruit. Transgenic Research 13, 373–384.
Das, P.K., Geul, B., Choi, S.B., Yoo, S.D., and Park, Y.I. 2011. Photosynthesis-
dependent anthocyanin pigmentation in Arabidopsis. Plant Signal
Behaviour 6, 23–25.
Di Carli, M., Zamboni, A., Pe, M.E., Pezzotti, M., Lilley, K.S., Benvenuto, E.,
and Desiderio, A. 2011. Two-dimensional differential in gel electropho-
resis (2D-DIGE) analysis of grape berry proteome during postharvest
withering. Journal of Proteome Research 10, 429–446.
Eikmanns, B.J., Follettie, M.T., Griot, M.U., and Sinskey, A.J. 1989. The phos-
phoenolpyruvate carboxylase gene of Corynebacterium glutamicum:
molecular cloning, nucleotide sequence, and expression. Molecular and
General Genetics 218, 330–339.
Esteve, C., D’Amato, A., Marina, M.L., Garcia, M.C., and Righetti, P.G. 2013.
In-depth proteomic analysis of banana (Musa spp.) fruit with combinato-
rial peptide ligand libraries. Electrophoresis 34, 207–214.
Etienne, A., Génard, M., Lobit, P., Mbeguie-A-Mbeguie, D., and Bugaud, C.
2013. What controls fleshy fruit acidity? A review of malate and citrate
accumulation in fruit cells. Journal of Experimental Botany, doi: 10.1093/
jxb/ert035.
FAOSTAT. 2013. faostat.fao.org/site/339/default.aspx (accessed October
2014).
Fei, Z., Tang, X., Alba, R.M., White, J.A., Ronning, C.M., Martin, G.B.,
Tanksley, S.D., and Giovannoni, J.J. 2004. Comprehensive EST analysis
of tomato and comparative genomics of fruit ripening. Plant Journal 40,
47–59.
Ferreyra, M.L.F., Rius, S.P., and Casati, P. 2012. Flavonoids: Biosynthesis, bio-
logical functions, and biotechnological applications. Frontiers in Plant
Science, doi: 10.3389/fpls.2012.00222.
Fischer, R.L., and Bennett, A.B. 1991. Role of cell-wall hydrolases in fruit rip-
ening. Annual Review of Plant Physiology and Plant Molecular Biology
42, 675–703.
Flores, F.B., Martinez-Madrid, M.C., Sanchez-Hidalgo, F.J., and Romojaro, F.
2001. Differential rind and pulp ripening of transgenic antisense ACC
oxidase melon. Plant Physiology and Biochemistry 39, 37–43.
Flügge, U.-I. 1999. Phosphate translocators in plastids. Annual Review of
Plant Physiology and Plant Molecular Biology 50, 27–45.
Gauci, V.J., Wright, E.P., and Coorssen, J.R. 2011. Quantitative proteomics:
assessing the spectrum of in-gel protein detection methods. Journal of
Chemical Biology 4, 3–29.

441
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Giavalisco, P., Nordhoff, E., Lehrach, H., Gobom, J., and Klose, J. 2003.
Extraction of proteins from plant tissues for two-dimensional electro-
phoresis analysis. Electrophoresis 24, 207–216.
Giovannoni, J. 2001. Molecular biology of fruit maturation and ripening.
Annual Review of Plant Physiology and Plant Molecular Biology 52,
725–749.
Giribaldi, M., Perugini, I., Sauvage, F.X., and Schubert, A. 2007. Analysis of
protein changes during grape berry ripening by 2-DE and MALDI-TOF.
Proteomics 7, 3154–3170.
Good, X., Kellogg, J.A., Wagoner, W., Langhoff, D., Matsumura, W., and
Bestwick, R.K. 1994. Reduced ethylene synthesis by transgenic toma-
toes expressing S-adenosylmethionine hydrolase. Plant Molecular
Biology 26, 781–790.
Gotz, S., Arnold, R., Sebastian-Leon, P., Martin-Rodriguez, S., Tischler, P.,
Jehl, M.A., Dopazo, J., Rattei, T., and Conesa, A. 2011. B2G-FAR, a spe-
cies-centered GO annotation repository. Bioinformatics 27, 919–924.
Gotz, S., Garcia-Gomez, J.M., Terol, J., Williams, T.D., Nagaraj, S.H., Nueda,
M.J., Robles, M., Talon, M., Dopazo, J., and Conesa, A. 2008. High-
throughput functional annotation and data mining with the Blast2GO
suite. Nucleic Acids Research 36, 3420–3435.
Grassi, S., Piro, G., Lee, J.M., Zheng, Y., Fei, Z., Dalessandro, G., Giovannoni,
J.J., and Lenucci, M.S. 2013. Comparative genomics reveals candidate
carotenoid pathway regulators of ripening watermelon fruit. BMC
Genomics 14, 1–20.
Greenbaum, D., Colangelo, C., Williams, K., and Gerstein, M. 2003. Comparing
protein abundance and mRNA expression levels on a genomic scale.
Genome Biology 4, 117.
Guarino, C., Arena, S., De Simone, L., D’Ambrosio, C., Santoro, S., Rocco,
M., Scaloni, A., and Marra, M. 2007. Proteomic analysis of the major
soluble components in Annurca apple flesh. Molecular Nutrition and
Food Research 51, 255–262.
Gygi, S.P., Rochon, Y., Franza, B.R., and Aebersold, R. 1999. Correlation
between protein and mRNA abundance in yeast. Molecular and Cellular
Biology 19, 1720–1730.
Hamilton, A.J., Lycett, G.W., and Grierson, D. 1990. Antisense gene that
inhibits synthesis of the hormone ethylene in transgenic plants. Nature
346, 284–287.
Hatzimanikatis, V., Choe, L.H., and Lee, K.H. 1999. Proteomics: Theoretical
and experimental considerations. Biotechnology Progress 15, 312–318.
Heazlewood, J.L., and Millar, A.H. 2003. Integrated plant proteomics—
Putting the green genomes to work. Functional Plant Biology 30,
471–482.
Heller, M.J. 2002. DNA microarray technology: Devices, systems, and appli-
cations. Annual Review of Biomedical Engineering 4, 129–153.

442
Proteomics of Fruit Development and Ripening

Hiwasa, K., Kinugasa, Y., Amano, S., Hashimoto, A., Nakano, R., Inaba, A.,
and Kubo, Y. 2003. Ethylene is required for both the initiation and
progression of softening in pear (Pyrus communis L.) fruit. Journal of
Experimental Botany 54, 771–779.
Honda, C., Kotoda, N., Wada, M., Kondo, S., Kobayashi, S., Soejima, J., Zhang,
Z., Tsuda, T., and Moriguchi, T. 2002. Anthocyanin biosynthetic genes
are coordinately expressed during red coloration in apple skin. Plant
Physiology and Biochemistry 40, 955–962.
Hurkman, W.J., and Tanaka, C.K. 1986. Solubilization of plant membrane-
proteins for analysis by two-dimensional gel-electrophoresis. Plant
Physiology 81, 802–806.
Iannetta, P.P.M., Laarhoven, L.J., Medina-Escobar, N., James, E.K., McManus,
M.T., Davies, H.V., and Harren, F.J.M. 2006. Ethylene and carbon dioxide
production by developing strawberries show a correlative pattern that is
indicative of ripening climacteric fruit. Physiologia Plantarum 127, 247–259.
Isaacson, T., Damasceno, C.M.B., Saravanan, R.S., He, Y., Catala, C., Saladie,
M., and Rose, J.K. 2006. Sample extraction techniques for enhanced
proteomic analysis of plant tissues. Nature Protocols 1, 769–774.
Ito, Y., Kitagawa, M., Ihashi, N., Yabe, K., Kimbara, J., Yasuda, J., Ito, H.,
Inakuma, T., Hiroi, S., and Kasumi, T. 2008. DNA-binding specificity,
transcriptional activation potential, and the rin mutation effect for the
tomato fruit-ripening regulator RIN. Plant Journal 55, 212–223.
Jia, H.F., Chai, Y.M. Li, C.L., Lu, D., Luo, J.J., Qin, L., and Shen, Y.Y. 2011.
Abscisic acid plays an important role in the regulation of strawberry
fruit ripening. Plant Physiology 157, 188–199.
Kambiranda, D., Katam, R., Basha, S.M., and Siebert, S. 2014. iTRAQ-based
quantitative proteomics of developing and ripening muscadine grape
berry. Journal of Proteome Research 13, 555–569.
Katz, E., Boo, K.H., Kim, H.Y., Eigenheer, R.A., Phinney, B.S., Shulaev, V., Negre-
Zakharov, F., Sadka, A., and Blumwald, E. 2011. Label-free shotgun pro-
teomics and metabolite analysis reveal a significant metabolic shift during
citrus fruit development. Journal of Experimental Botany 62, 5367–5384.
Katz, E., Fon, M., Eigenheer, R.A., Phinney, B.S., Fass, J.N., Lin, D., Sadka, A.,
and Blumwald, E. 2010. A label-free differential quantitative mass spec-
trometry method for the characterization and identification of protein
changes during citrus fruit development. Proteome Science 8, 68. doi:
10.1186/1477-5956-8-68.
Kimura, S., and Sinha, N. 2008. Tomato (Solanum lycopersicum): A model fruit-
bearing crop. Cold Spring Harbor Protocols, doi: 10.1101/pbd.emo105.
Klose, J. 1999. Genotypes and phenotypes. Electrophoresis 20, 643–652.
Laurent, J.M., Vogel, C., Kwon, T., Craig, S.A., Boutz, D.R., Huse, H.K.,
Nozue, K., Walia, H., Whiteley, M., Ronald, P.C., and Marcotte, E.M.
2010. Protein abundances are more conserved than mRNA abundances
across diverse taxa. Proteomics 10, 4209–4212.

443
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lee, J.M., Kim, S., Lee, J.Y., Yoo, E.Y., Cho, M.C., Cho, M.R., Kim, B.D., and
Bahk, Y.Y. 2006. A differentially expressed proteomic analysis in placen-
tal tissues in relation to pungency during the pepper fruit development.
Proteomics 6, 5248–5259.
Li, M.J., Feng, F.J., and Cheng, L.L. 2012. Expression patterns of genes
involved in sugar metabolism and accumulation during apple fruit
development. PLoS One 7.
Lokkamlue, N., and Huehne, P.S. 2013. Cloning and sequence of cDNA
encoding 1-aminocyclo-propane-1-carboxylate oxidase in Vanda flowers.
Maejo International Journal of Science and Technology 7, 338–352.
Lopez-Gomez, R., Cabrera-Ponce, J.L., Saucedo-Arias, L.J., Carreto-Montoya,
L., Villanueva-Arce, R., Diaz-Perez, J.C., Gomez-Lim, M.A., and Herrera-
Estrella, L. 2009. Ripening in papaya fruit is altered by ACC oxidase
cosuppression. Transgenic Research 18, 89–97.
Mann, M., and Jensen, O.N. 2003. Proteomic analysis of post-translational
modifications. Nature Biotechnology 21, 255–261.
Marondedze, C. 2011. Fruit Quality Traits in Apple. Lambert Academic,
Saarbrücken, Germany.
Marondedze, C., and Thomas, L.A. 2012a. Insights into fruit function from
the proteome of the hypanthium. Journal of Plant Physiology 169, 12–19.
Marondedze, C., and Thomas, L.A. 2012b. Apple hypanthium firmness:
New insights from comparative proteomics. Applied Biochemistry and
Biotechnology 168, 306–326.
Marondedze, C., Gehring, C., and Thomas, L. 2014. Dynamic changes in the
date palm fruit proteome during development and ripening. Horticulture
Research 1, 14039. doi: 10.1038/hortres.2014.39.
Martinez-Esteso, M.J., Selles-Marchart, S., Lijavetzky, D., Angeles-Pedreno,
M., and Bru-Martinez, R. 2011. A DIGE-based quantitative proteomic
analysis of grape berry flesh development and ripening reveals key
events in sugar and organic acid metabolism. Journal of Experimental
Botany 62, 2521–2569.
Mayer, A.M. 2006. Polyphenol oxidases in plants and fungi: going places? A
review. Phytochemistry 67, 2318–2331.
Merzlyak, M.N., and Chivkunova, O.B. 2000. Light-stress-induced pigment
changes and evidence for anthocyanin photoprotection in apples.
Journal of Photochemistry and Photobiology B: Biology 55, 155–163.
Metzker, M.L. 2010. Sequencing technologies—The next generation. Nature
Review Genetics 11, 31–46.
Molassiotis, A., Tanou, G., Filippou, P., and Fotopoulos, V. 2013. Proteomics
in the fruit tree science arena: New insights into fruit defense, develop-
ment, and ripening. Proteomics 13, 1871–1884.
Muccilli, V., Licciardello, C., Fontanini, D., Russo, M.R., Cunsolo, V., Saletti,
R., Recupero, G.R., and Foti, S. 2009. Proteome analysis of Citrus
sinensis L. (Osbeck) flesh at ripening time. Journal of Proteomics 73,
134–152.

444
Proteomics of Fruit Development and Ripening

Nascimento, N.C., and Fett-Neto, A.G. 2010. Plant secondary metabolism

and challenges in modifying its operation: an overview. Methods in
Molecular Biology 643, 1–13.
Nergiz, C., and Ergönül, P.G. 2009. Organic acid content and composition of
the olive fruits during ripening and its relationship with oil and sugar.
Scientia Horticulturae 122, 216–220.
Nogueira, S.B., Labate, C.A., Gozzo, F.C., Pilau, E.J., Lajolo, F.M., and do
Nascimento, J.R.O. 2012. Proteomic analysis of papaya fruit ripening
using 2DE-DIGE. Journal of Proteomics 75, 1428–1439.
Oeller, P.W., Wong, L.M., Taylor, L.P., Pike, D.A., and Theologis, A. 1991.
Reversible inhibition of tomato fruit senescence by antisense RNA.
Science 254, 437–439.
Old, W.M., Meyer-Arendt, K., Aveline-Wolf, L., Pierce, K.G., Mendodza, A.,
Sevinsky, J.R., Resing, K.A., and Ahn, N.G. 2005. Comparison of label-
free methods for quantifying human proteins by shotgun proteomics.
Molecular and Cellular Proteomics 4, 1487–1502.
Orellana, A., and Nilo, R. 2012. Fruit proteomics. In Proteomic Applications in
Biology, ed. Heazlewood, J. InTech. Rijeka, Croatia, 33–53.
Palma, J.M., Corpas, F.J., and del Rio, L.A. 2011. Proteomics as an approach
to the understanding of the molecular physiology of fruit development
and ripening. Journal of Proteomics 74, 1230–1243.
Payasi, A., Mishra, N.N., Chaves, A.L.S., and Singh, R. 2009. Biochemistry of
fruit softening: An overview. Physiology and Molecular Biology of Plants
15, 103–113.
Pedreschi, R., Vanstreels, E., Carpentier, S., Hertog, M., Lammertyn, J.,
Robben, J., Noben, J.P., Swennen, R., Vanderleyden, J., and Nicolai, B.M.
2007. Proteomic analysis of core breakdown disorder in Conference
pears (Pyrus communis L.). Proteomics 7, 2083–2099.
Pham, T.N.T. 2007. Ripening behaviour of capsicum (Capsicum annuum L.)
fruit. PhD thesis, University of Adelaide, Australia.
Prinsi, B., Negri, A.S., Fedeli, C., Morgutti, S., Negrini, N., Cocucci, M., and
Espen, L. 2011. Peach fruit ripening: a proteomic comparative analysis
of the mesocarp of two cultivars with different flesh firmness at two
ripening stages. Phytochemistry 72, 1251–1262.
Rambla, J.L., Tikunov, Y.M., Monforte, A.J., Bovy, A.G., and Granell, A. 2014.
The expanded tomato fruit volatile landscape. Journal of Experimental
Botany 65, 4613–4623.
Riederer, B.M. 2008. Non-covalent and covalent protein labeling in two-
dimensional gel electrophoresis. Journal of Proteomics 71, 231–244.
Rocco, M., D’Ambrosio, C., Arena, S., Faurobert, M., Scaloni, A., and Marra,
M. 2006. Proteomic analysis of tomato fruits from two ecotypes during
ripening. Proteomics 6, 3781–3791.
Rose, J.K.C., Bashir, S., Giovannoni, J.J., Jahn, M.M., and Saravanan, R.S.
2004. Tackling the plant proteome: Practical approaches, hurdles and
experimental tools. Plant Journal 39, 715–733.

445
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Ruf, S., Hermann, M., Berger, I.J., Carrer, H., and Bock, R. 2001. Stable
genetic transformation of tomato plastids and expression of a foreign
protein in fruit. Nature Biotechnology 19, 870–875.
Saito, T., Tohru, A., Yoshihiro, O., Asamizu, E., Hiwasa-Tanase, K., Fukuda, N.,
Mizoguchi, T., Yamazaki, Y., Aoki, K., and Ezura, H. 2011. TOMATOMA:
A novel tomato mutant database distributing micro-tom mutant collec-
tions. Plant and Cell Physiology 52, 283–296.
Santoni, V., Doumas, P., Rouquie, D., Mansion, M., Rabilloud, T., and
Rossignol, M. 1999. Large scale characterization of plant plasma mem-
brane proteins. Biochimie 81, 655–661.
Santoni, V., Molloy, M., and Rabilloud, T. 2000. Membrane proteins and pro-
teomics: Un amour impossible? Electrophoresis 21, 1054–1070.
Saravanan, R.S., and Rose, J.K. 2004. A critical evaluation of sample extrac-
tion techniques for enhanced proteomic analysis of recalcitrant plant
tissues. Proteomics 4, 2522–2532.
Sarry, J.E., Sommerer, N., Sauvage, F.X., Bergoin, A., Rossignol, M., Albagnac,
G., and Romieu, C. 2004. Grape berry biochemistry revisited upon pro-
teomic analysis of the mesocarp. Proteomics 4, 201–215.
Shaw, M.M., and Riederer, B.M. 2003. Sample preparation for two-dimen-
sional gel electrophoresis. Proteomics 3, 1408–1417.
Shi, Y., Jiang, L., Zhang, L., Kang, R., and Yu, Z. 2014. Dynamic changes in
proteins during apple (Malus domestica) fruit ripening and storage.
Horticulture Research 6. doi:10.1038/hortres.2014.6.
Silva, J.C., Denny, R., Dorschel, C.A., Gorenstein, M., Kass, I.J., Li, G.Z.,
McKenna, T., Nold, M.J., Richardson, K., Young, P., and Geromanos, S.
2005. Quantitative proteomic analysis by accurate mass retention time
pairs. Analytical Chemistry 77, 2187–2200.
Smillie, R.M., and Hetherington, S.E. 1999. Photoabatement by anthocyanin
shields photosynthetic systems from light stress. Photosynthetica 36,
451–463.
Sun, L.A., Zhang, M., Ren, J., Qi, J., Zhang, G., and Leng, P. 2010.
Reciprocity between abscisic acid and ethylene at the onset of
berry ripening and after harvest. BMC Plant Biology 10, 257. doi:
10.1186/1471-2229-10-257.
Symons, G.M., Davies, C., Shavrukov, Y., Dry, I.B., Reid, J.B., and Thomas,
M.R. 2006. Grapes on steroids. Brassinosteroids are involved in grape
berry ripening. Plant Physiology 140, 150–158.
Talke, I.N., Blaudez, D., Maathuis, F.J.M., and Sanders, D. 2003. CNGCs:
prime targets of plant cyclic nucleotide signalling? Trends in Plant
Science 8, 286–293.
Teixeira, A., Eiras-Dias, J., Castellarin, S.D., and Geros, H. 2013. Berry pheno-
lics of grapevine under challenging environments. International Journal
of Molecular Sciences 14, 18711–18739.
Thumdee, S., Manenoi, A., and Paull, R.E. 2007. Activity of papaya fruit
hydrolases during natural softening and modified softening. Acta
Horticulturae 740, 317–322.

446
Proteomics of Fruit Development and Ripening

Toledo, T.T., Nogueira, S.B., Cordenunsi, B.R., Gozzo, F.C., Pilau, E.J., Lajolo,
F.M., and do Nascimento, J.R.O. 2012. Proteomic analysis of banana
fruit reveals proteins that are differentially accumulated during ripen-
ing. Postharvest Biology and Technology 70, 51–58.
Tomato Genome Consortium. 2012. The tomato genome sequence provides
insights into fleshy fruit evolution. Nature 485, 635–641.
Toorchi, M., Nouri, M.Z., Tsumura, M., and Komatsu, S. 2008. Acoustic tech-
nology for high-performance disruption and extraction of plant pro-
teins. Journal of Proteome Research 7, 3035–3041.
Trainotti, L., Pavanello, A., and Casadoro, G. 2005. Different ethylene recep-
tors show an increased expression during the ripening of strawberries:
does such an increment imply a role for ethylene in the ripening of these
non-climacteric fruits? Journal of Experimental Botany 56, 2037–2046.
Tretter, L., and Adam-Vizi, V. 2005. Alpha-ketoglutarate dehydrogenase: a tar-
get and generator of oxidative stress. Philosophical Transactions of the
Royal Society B: Biological Sciences 360, 2335–2345.
Vander Mijnsbrugge, K., Meyermans, H., Montagu, M.V., Bauw, G., and
Boerjan, W. 2000. Wood formation in poplar: Identification, characteriza-
tion, and seasonal variation of xylem proteins. Planta 210, 589–598.
Vincent, D., Wheatley, M.D., and Cramer, G.R. 2006. Optimization of pro-
tein extraction and solubilization for mature grape berry clusters.
Electrophoresis 27, 1853–1865.
Wang, W., Scali, M., Vignani, R., Spadafora, A., Sensi, E., Mazzuca, S., and
Cresti, M. 2003. Protein extraction for two-dimensional electrophoresis
from olive leaf, a plant tissue containing high levels of interfering com-
pounds. Electrophoresis 24, 2369–2375.
Wilkinson, J.Q., Lanahan, M.B., Clark, D.G., Bleecker, A.B., Chang, C.,
Meyerowitz, E.M., and Klee, H.J. 1997. A dominant mutant receptor
from Arabidopsis confers ethylene insensitivity in heterologous plants.
Nature Biotechnology 15, 444–447.
Williams, R.J., Spencer, J.P., and Rice-Evans, C. 2004. Flavonoids: antioxidants
or signalling molecules? Free Radical Biology and Medicine 36, 838–849.
Wu, H.X., Jia, H.M., Ma, X.W., Wang, S.B., Yao, Q.S., Xu, W.T., Zhou, Y.G.,
Gao, Z.S., and Zhan, R.L. 2014. Transcriptome and proteomic analy-
sis of mango (Mangifera indica Linn) fruits. Journal of Proteomics 105,
19–30.
Wu, X., Gong, F., and Wang, W. 2014a. Protein extraction from plant tis-
sues for 2DE and its application in proteomic analysis. Proteomics 14,
645–658.
Wu, X., Xiong, E., Wang, W., Scali, M., and Cresti, M. 2014b. Universal sample
preparation method integrating trichloroacetic acid/acetone precipita-
tion with phenol extraction for crop proteomic analysis. Nature Protocol
9, 362–374.
Xu, J., Pascual, L., Aurand, R., Bouchet, J.P., Valot, B., Zivy, M., Causse,
M., and Faurobert, M. 2013. An extensive proteome map of tomato
(Solanum lycopersicum) fruit pericarp. Proteomics 13, 3059–3063.

447
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Zelman, A.K., Dawe, A., Gehring, C., and Berkowitz, G.A. 2012. Evolutionary
and structural perspectives of plant cyclic nucleotide gated cation chan-
nels. Frontiers in Plant Science 3, 95. doi: 10.3389/fpls.2012.00095.
Zhang, J., Ma, H., Feng, J., Zheng, L., Wang, Z., and Chen, S. 2008. Grape
berry plasma membrane proteome analysis and its differential expres-
sion during ripening. Journal of Experimental Botany 59, 2979–2990.
Zhang, Y., Butelli, E., De Stefano, R., Schoonbeek, H.J., Magusin, A.,
Pagaliarani, C., Wellner, N., Hill, L., Orzaez, D., Granel, A., Jones, J.D.G.,
and Martin, C. 2013. Anthocyanins double the shelf life of tomatoes by
delaying overripening and reducing susceptibility to gray mold. Current
Biology 23, 1094–1100.
Zheng, Q., Song, J., Doncaster, K., Rowland, E., and Byers, D.M. 2007.
Qualitative and quantitative evaluation of protein extraction protocols
for apple and strawberry fruit suitable for two-dimensional electropho-
resis and mass spectrometry analysis. Journal of Agricultural and Food
Chemistry 55, 1663–1673.

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Chapter 14

Potato Tuber Dormancy

and Postharvest
Sprout Control
Jeffrey C. Suttle,1 Michael A. Campbell,2
and Nora L. Olsen3
1USDA-ARS Northern Crop Science Laboratory,
Fargo, North Dakota
2 Penn State Erie, The Behrend College, Erie, Pennsylvania
3 University of Idaho, Twin Falls, Idaho

Abstract 450
14.1 Introduction 451
14.2 Dormancy: General Considerations 451
14.2.1  Developmental Aspects 452
14.3 Genetics of Tuber Dormancy 453
14.4 Pre- and Postharvest Environmental Effects 453
14.5 Physiological Regulation of Tuber Dormancy 454
14.5.1 Auxin 455
14.5.2 Carotenoid-Derived Hormones: Abscisic
Acid and Strigolactone 456
14.5.3 Cytokinins 458
14.5.4 Ethylene 459

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14.5.5 Gibberellins 460
14.5.6  Summary and Conclusions 460
14.6 Transcriptional Regulation during Dormancy 461
14.6.1  Initiation of Tuber Dormancy 461
14.6.2  Dormancy Termination 462
14.6.3  Epigenetic Control of Tuber Dormancy 463
14.7 Control of Sprouting in Storage 463
14.7.1  Introduction and Importance 463
14.7.2  Dormancy, Cultivar Selection, and
Storage Temperature 464
14.7.3  Chemical Control 464
14.8 Conclusions and Future Perspectives 467
References 467

Abstract
Potato is the third most important food crop in the world and is an excel-
lent source of dietary calories, vitamins, and minerals. At harvest and for an
indeterminate period thereafter, potato tubers are in a state of physiological
dormancy and will not sprout. Tuber dormancy is lost during storage in a cul-
tivar- and environmentally dependent manner. The onset of sprouting, which
follows the loss of tuber dormancy, results in numerous biochemical changes
that are detrimental to the processing and nutritional qualities of potatoes.
The initiation of tuber dormancy is coincident with the initiation of
tuberization. The duration of tuber dormancy is affected by both genetic and
pre- and postharvest environmental factors. Endogenous plant hormones
are involved in the regulation of tuber dormancy progression. Current evi-
dence suggests that abscisic acid and ethylene are required for dormancy
initiation and maintenance, while cytokinins appear to regulate dormancy
exit. The exact roles of auxins and gibberellins in tuber dormancy control
have yet to be determined, but both are involved in the control of sprout
growth that follows dormancy termination.
Dormancy initiation and progression are associated with wholesale
changes in gene expression. Although the transcript abundances of many
genes change dramatically during tuber dormancy, in most cases, the bio-
logical significance of these changes has yet to be determined. Dormant
tuber meristems are arrested in the G1 position of the cell cycle, and the
onset of sprout growth following dormancy exit is accompanied by the
resumption of cell cycle progression and expression of genes encoding key
proteins involved in cell cycle control. Dormancy progression is also accom-
panied by changes in chromatin composition involving both DNA cytosine
methylation and covalent histone modification.

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Because of its deleterious effects on tuber quality, successful long-

term storage of potatoes requires the control of sprouting. In most commer-
cial storages, sprout control is achieved through the application of sprout
inhibitors. The most widely used sprout inhibitors are synthetic chemicals
that inhibit sprouting in an herbicidal manner. Several newer natural product-
based sprout inhibitors have been introduced whose mechanisms of action
are distinct from those of the earlier synthetic sprout inhibitors.

14.1 Introduction
Together with rice and cereals, potatoes are among the top three crops
grown worldwide for human consumption. Global potato production
exceeds 300  million metric tons, with China, India, and the Russian
Federation leading in total production. In addition to its contribution to
basic total caloric intake, potatoes are excellent sources of minerals, vita-
mins, and fiber. A single 150 g potato contains roughly 45% of the minimum
daily requirement of vitamin C and more potassium than any commonly
consumed vegetable or fruit.
In the United States, potatoes are grown on more than 1.1 million
acres, and in 2012, more than 46.7 billion pounds of potatoes were har-
vested, with an estimated market value in excess of $3.9 billion (USDA/
NASS, 2013). More than 70% of the U.S. fall potato production is placed into
medium- to long-term storage to satisfy the yearlong demands of both the
processing industry and consumers. Unlike rice and cereals, potatoes are
stored in a fully hydrated and highly perishable form. Maintenance of the
potato market and nutritional qualities during storage is a critical aspect of
modern potato storage management practices.
In addition to postharvest disease, uncontrolled sprouting results in
accelerated loss of tuber quality. At harvest, and for an indeterminate period
thereafter, potato tubers are dormant and will not sprout. During extended
postharvest storage, tuber dormancy is gradually lost and sprout growth
commences. Accompanying sprout growth is a host of biochemical changes
that severely impact both the nutritional and processing qualities of stored
tubers. In addition, the increased water loss from sprouting tubers predis-
poses the tubers to accelerated pathogen attack. For these reasons, control of
postharvest sprouting is an important aspect of potato storage management.

14.2  Dormancy: General Considerations

All organisms from prokaryotes to complex eukaryotic organisms have
evolved mechanisms to cope with environmental stresses such as drought
and temperature extremes. In general, organisms can either avoid the stress

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by migrating or endure the stress through biochemical or developmental

means. Dormancy or metabolic quiescence is one physiological strategy
that has successfully been employed to survive environmental extremes.
Because of their sessile growth habit, plants have evolved a variety of forms
of dormancy, including seed and bud dormancy.
In its broadest terms, dormancy is defined by the absence of visible
growth (Samish, 1954). Three types of plant dormancy have been catego-
rized based on the origin of the factors inhibiting growth (Lang et al., 1987).
Endodormancy arises from factors originating from within the affected
meristem. Paradormancy is induced by endogenous factors originating out-
side the affected meristem, and ecodormancy occurs when external envi-
ronmental factors prevent growth. During a typical life cycle, potato tuber
buds or meristems pass through each of these phases. At harvest and for an
indeterminate period thereafter, tuber meristems are in a state of endodor-
mancy and will not sprout. As endodormancy weakens, one or more tuber
buds initiates growth, becomes dominant, and inhibits the growth of other
buds on the same tuber (paradormancy). If at this time tubers are in an
unfavorable environment with suboptimal temperatures, such as a storage
bin, tuber buds will not sprout because of ecodormancy. Under this scheme,
potato tuber buds whose growth has been arrested because of the applica-
tion of a sprout inhibitor (even if the effects are only temporary) are not
considered dormant.

14.2.1  Developmental Aspects

From a developmental perspective, potato tuber growth is initiated when the
longitudinal growth of the stolon ceases and subapical swelling commences
(Xu et al., 1998b). Photoperiod and certain mobile signals, including the
potato Flowering Locus T homolog StSP6A and possibly low-­molecular-
weight hormones, have been implicated in this process (Kloosterman
et al., 2007; Navarro et al., 2011). Thus, tuberization is initiated by shoot-
imposed growth cessation (paradormancy) of the stolon apical meristem.
Supporting this hypothesis, extensive studies by Burton (1989) found that
the length of tuber dormancy in cultivars exhibiting a wide range of dor-
mant periods was correlated with the timing of tuber initiation rather than
harvest date.
Potato tubers are determinate organs with a finite life span. The term
physiological aging is used to describe the entire life cycle of a tuber, begin-
ning with initiation and ultimately ending in senescence and the inability
to sprout (Coleman, 2000). Thus, tuber dormancy is only part of the devel-
opmental continuum that physiologically overlaps initially with initiation
and later with vigorous sprout growth and likely shares common regulatory
mechanisms with these processes.

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14.3  Genetics of Tuber Dormancy

Early studies by Simmonds (1964) demonstrated that tuber dormancy
duration had a strong genetic component and was likely under polygenic
control. Direct genetic control of potato tuber dormancy was first demon-
strated using quantitative trait locus (QTL) analysis. Twenty-two markers
were shown to be associated with segregation of the length of dormancy
in a diploid population of hybrid potato (Solanum tuberosum × Solanum
chacoense) crossed with Solanum phureja (Freyre et al., 1994). Additional
studies using QTL analysis on reciprocal backcrosses of S. tuberosum and
Solanum berthaultii also supported a genetic link to the control of dormancy
in potato (van den Berg et al., 1996). Although QTL analysis did not lead to
specific genes, it did reveal that there is a genetic component to the regula-
tion of potato dormancy. However, QTL analysis did reveal an association
between high abscisic acid (ABA) content and long tuber dormancy (Šimko
et al., 1997). More recent studies using transcriptional analysis, via micro-
arrays or RNA-seq, have supported a strong link between genetic regula-
tion and dormancy status.
Interestingly, the early studies of Simmonds (1964) examining the
genetic component of tuber dormancy control also revealed a strong posi-
tive correlation between tuber and true seed dormancies such that selection
for one effectively selects for the other. This property likely explains the
relatively short-dormancy duration of many modern cultivars as the rapid
germination of true seed from initial sexual crosses in the greenhouse has
inadvertently selected for reduced dormancy in true seed that forms the
basis of most potato breeding programs.

14.4  Pre- and Postharvest Environmental Effects

Unlike the marked effect of genetics on the length of tuber dormancy, within
nominal constraints, preharvest environmental conditions have little overall
effect on the depth or duration of the dormant period. Exhaustive studies by
Emilsson (1949) found little effect of day length on the duration of tuber dor-
mancy. Similarly, nominal variations in temperature during tuber growth
have only a limited effect on the subsequent length of tuber dormancy
(Schippers, 1956). However, if the growing season is unusually hot, dry, or
cold, there can be a profound effect on tuber dormancy duration. For exam-
ple, abnormally cool or hot temperatures during the growing season were
found to increase or decrease the period of dormancy, respectively (Burton,
1963). In particular, sustained nighttime temperatures of >32°C result in
resumption of apical stolon elongation and cessation of tuber growth, caus-
ing what is commonly referred to as heat sprouts (Bodlaender et al., 1964).
With the return to cooler night temperatures, stolon elongation ceases and

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tuber growth resumes or is reinitiated, resulting in either severe tuber mal-

formation or chain tuberization (Hiller et al., 1985).
Postharvest temperatures (typically in a commercial storage) have a
much greater impact on tuber dormancy duration. Between the tempera-
tures of 4°C and 20°C, tuber dormancy duration varies inversely with tem-
perature (for review, see Burton, 1989). Exposure to temperatures above
or below this range can rapidly result in the abrupt termination of tuber
dormancy and the onset of sprout growth. Unfortunately, the optimum
storage temperatures for tubers destined for processing (ca. 7°C) are well
above that needed to significantly retard or prevent dormancy release and
sprout growth, thus necessitating the application of some form of artificial
sprout control.
In addition to temperature, storage atmosphere can affect tuber dor-
mancy and subsequent sprout growth. In particular, hypoxic conditions
(i.e., [O2] ca. 2%–4%) can effectively break tuber dormancy and result in
sprouting when normal oxygen content is restored. Interestingly, elevated
CO2 concentrations do not appear to affect tuber dormancy. In addition to
respiratory gases, tubers produce a number of volatile compounds, some of
which have been found to influence tuber dormancy and sprout growth. Low
amounts of ethylene are produced constitutively by potato tubers, and the
rate of production increases dramatically after wounding or pathogen attack
(Okazawa, 1974). Short-term exposure to relatively high concentrations
of ethylene has been reported to break tuber dormancy, and continuous
exposure to lower levels suppresses sprout growth (Rylski et al., 1974). In
addition to ethylene, tubers produce a range of aromatic and aliphatic hydro-
carbons, some of which have been shown to inhibit sprout growth (Burton
and Meigh, 1971; Meigh et al., 1973). Notably, the sprout growth–inhibiting
activities of ethylene and the potato volatile 1,4-dimethylnaphthalene have
been exploited commercially.

14.5  Physiological Regulation of Tuber Dormancy

As indicated above, potato tuber bud dormancy is thought to begin at tuber-
ization and continues for an indeterminate period of time after harvest.
Although the exact molecular processes controlling entry into and exit
from dormancy are unknown, endogenous hormones have been demon-
strated to participate in the regulation of tuber bud dormancy progression
(Figure 14.1). In this section, the putative role for endogenous hormones
in dormancy regulation is discussed with emphasis on more recent results
employing physiochemical methods of hormone analysis. In assessing the
possible roles of endogenous hormones, three criteria will be examined:
(1) changes in endogenous content, (2) chemical or genetic manipulation of
endogenous levels, and (3) effects of exogenous application.

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 P OTATO TUBER D ORMAN C Y

Short Days Sprouting

Tuber Initiation
Ecodormant
Endodormant
Deep Dormancy

FT-homologs Cell division

associated transcripts
ABA-induced
Histone
genes
expression
Dormancy-associated UTPase
transcripts
GA
DAM increase
MADS-box
Symplastic
ABA isolation of ABA Cytokinin
increase meristem decrease increase

Figure 14.1  Schematic representation of the timeline of tuber dormancy pro-

gression beginning with tuber initiation (left) and progressing to the onset of
sprouting (right). Concurrent with tuber initiation is the initiation of endodor-
mancy, which progresses to ecodormancy and ultimately to sprouting. Major
changes in gene expression are indicated along this time axis, as are the
principal changes in endogenous hormone contents.

14.5.1 Auxin
Despite being the first plant hormone described, there have been relatively few
detailed analyses of changes in endogenous auxins (notably Indole-3-acetic
acid (IAA) and its metabolites) during tuber dormancy progression, and as
such, their role in the regulation of potato tuber bud dormancy remains uncer-
tain. As determined by enzyme-linked immunosorbent assay (ELISA), endog-
enous IAA content in tuber apical buds remained essentially constant during 5
months of postharvest storage and declined markedly as sprouting commenced
(Sukhova et  al., 1993). Subsequent studies using gas chromatography–mass
spectrometry (GC-MS) analysis of tuber apical bud extracts by Sorce et  al.
(2000, 2009) provided conflicting data. In the first study, the content of IAA
increased dramatically during postharvest storage, peaking coincident with or
slightly after sprouting commenced. In the second study, using two cultivars
with differing lengths of innate dormancy, the endogenous content of IAA in
both cultivars was highest at harvest, declined during storage, and reached
a minimum as sprouting began. Consistent with the latter study, expression
of the auxin-inducible gene ARF6 was low in dormant buds and increased
as sprouting commenced (Faivre-Rampant et al., 2004), suggesting that an
increase in endogenous auxin content accompanies dormancy cessation.
Application of low concentrations of IAA to nondormant buds slightly
stimulates elongation but has no apparent effect on dormant buds (Hemberg,

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1985). Treatment with high concentrations (>100 µM) of s­ ynthetic auxins

such as α-naphthalene acetic acid inhibits the growth of buds on nondor-
mant tubers (Suttle, 2003).
The biosynthesis of IAA in most plant tissues is complex and often
involves multiple redundant pathways (Korasick et al., 2013). As such, inhi-
bition of IAA biosynthesis by either chemical or genetic means has not been
reported in any potato tissue. However, genetic manipulation of IAA con-
tent or the sensitivity in many plant species has been reported using either
the iaaM or iaaH genes of Agrobacterium tumefaciens or the RolB gene
of Agrobacterium rhizogenes (Klee et al., 1987). Transformation of potato
plantlets with the RolB gene resulted in a number of phenotypic alterations,
including tuber morphology (Schmülling et al., 1993). However, no mention
was made of any effects on tuber dormancy or sprout growth. The effects
of the A. tumefaciens auxin biosynthesis genes on tuber development have
not been reported.
Auxins have been posited to play a pivotal role in the initiation of seed
dormancy by modulating ABA signaling (Liu et al., 2013), and it would
be tempting to assign a similar role in tuber bud dormancy; however, evi-
dence gathered to date is inconsistent with that hypothesis. Clearly, more
research is needed before a definitive conclusion can be reached.

14.5.2 Carotenoid-Derived Hormones: Abscisic Acid
and Strigolactone
Prior to its isolation and chemical characterization, abscisic acid (ABA)
was known by two names, abscising and dormin, the latter arising from its
association with temperate perennial bud dormancy (Addicott and Carns,
1983). ABA was subsequently identified as a major bioactive component of
the β-inhibitor complex, whose levels (as judged by bioassay) were closely
correlated with several forms of meristem dormancy (Addicott and Carns,
1983; Hemberg, 1985). Studies using more selective and sensitive instru-
mental analyses have largely confirmed the early bioassay results and found
that tuber ABA content is highest in deeply dormant tuber tissues, declines
during postharvest storage, and is lowest at the time of sprouting (for
reviews, see Suttle, 2007; Sonnewald and Sonnewald, 2014). Unfortunately,
many of these studies used bulk tuber samples composed mainly of nonbud
tissue, thereby complicating interpretation of the data.
More recently, the effects of dormancy status on the ABA content
of isolated tuber tissues was determined using mass spectrometry. Using
GC-MS, Sorce et al. (1996) found the highest concentrations of ABA in
“subeye” tissues and also found a small increase in the ABA content of tuber
eyes as sprouting commenced. Subsequent LC-MS analyses examining
changes in the ABA content of microscopically excised tuber eyes during

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natural and chemically forced dormancy progression found a marked

decrease in ABA content prior to the onset of sprout growth (Destefano-
Beltrán et al., 2006a, 2006b). Consistent with these latter data, transcript
analyses of tuber meristem RNA during natural and chemically forced dor-
mancy progression demonstrated decreased expression of several ABA-
inducible genes (Campbell et al., 2008).
Exogenous ABA is rapidly metabolized to biologically inactive prod-
ucts in potato tuber tissues (Suttle et al., 2012). In addition, tuber sensitivity
to ABA appears to decline during the latter stages of dormancy. As a result,
the application of ABA to nondormant tubers elicits only a transient inhibi-
tion of sprout growth (El-Antably et al., 1967).
Demonstration of a role for endogenous ABA in tuber dormancy has
come from two lines of evidence: genetic and chemical. A mutant line of
S. phureja that exhibited a severe water-stressed phenotype even under
well-watered conditions and contained less than 10% of the ABA content
of a normal sib was identified and given the trivial name Droopy (Quarrie,
1982). Although Droopy plants will initiate tubers under short-day condi-
tions, because of precocious sprouting, the tubers are diminutive in size
and cannot be stored. The role of ABA in tuber dormancy was also assessed
using an in vitro microtuber system and the carotenoid biosynthesis inhibi-
tor fluridone (FLD). At harvest, after 9 weeks of development, microtubers
were deeply dormant and did not sprout for another 15–18 weeks (Suttle
and Hultstrand, 1994). However, microtubers generated on FLD-containing
medium contained <10% of the ABA content of control microtubers and
were completely nondormant at harvest. Inclusion of micrometer concen-
trations of ABA in FLD-containing medium completely prevented prema-
ture sprouting. Furthermore, treatment of harvested dormant microtubers
with FLD lowered endogenous ABA content and stimulated premature
sprouting. Interestingly, inhibition of ABA metabolism in dormant tubers
resulted in a sustained increase in ABA content but had no effect on dor-
mancy duration (Suttle et al., 2012). Collectively, these results suggest that
although endogenous ABA is essential for both the initiation and mainte-
nance of tuber dormancy, a decline in ABA content is not required for tuber
dormancy exit.
In addition to ABA, the oxidative cleavage of C 40 carotenoids yields
a variety of lower-molecular-weight compounds, some of which display
biological activity (Ohmiya, 2009). Strigolactones (SLs) are C18 carotenoid
derivatives that exhibit potent growth-regulating activities. In particular,
SLs have been shown to participate in the regulation of apical dominance
(paradormancy) by acting as root-derived inhibitors of axillary bud growth
(Dun et al., 2009). The initial steps of SL biosynthesis involve the actions of
two carotenoid cleavage dioxygenases, CCD7 and CCD8 (Ohmiya, 2009).
Recently, it has been shown that decreased expression of the potato CCD8
gene using RNAi results in a number of tuber abnormalities, including

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premature sprouting and excessive sprout branching (Pasare et al., 2013).

It is not clear if these effects are directly due to impaired SL biosynthesis or
are a result of alterations in other hormonal and regulatory systems.

14.5.3 Cytokinins
Cytokinins are known to participate in the control of cell division and the
resumption of cell cycle activity following the termination of meristematic
quiescence (Mok and Mok, 2001; Kyozuka, 2007). As such, they have been
implicated in the reinitiation of growth following dormancy exit in peren-
nial tree buds and potato tubers.
Increases in isoprenoid cytokinins, including both trans- and cis-
zeatin derivatives, have been detected in potato tuber buds as they exit dor-
mancy and initiate sprout growth (for reviews, see Suttle, 2007; Sonnewald
and Sonnewald, 2014). Exogenous cytokinins are rapidly degraded to
adenine derivatives in potato tubers by the enzyme cytokinin oxidase
(Suttle, 2002). However, no significant changes in cytokinin metabolism or
expression of cytokinin oxidase genes were observed during natural and
chemically forced dormancy progression (Suttle et al., 2014). These results
suggest that metabolic control of tuber cytokinin content during dormancy
occurs at the level of biosynthesis. Although genes encoding isopentenyl-
transferases (the first committed step in cytokinin biosynthesis) have been
cloned and characterized from a number of plants (Sakakibara, 2006), the
effect of tuber dormancy status on their expression has not been reported.
After harvest and during storage, tubers pass through three stages
of dormancy. Initially, tubers are deeply dormant and will not sprout in
response to a variety of external treatments. Next, tubers remain dormant
but become responsive to external agents/treatments. Finally, tuber dor-
mancy ends and sprouting commences naturally. The endogenous content
of cytokinins is low in phases 1 and 2, but increases coincident with the
onset of sprout growth.
As was first reported by Hemberg (1970), once tubers become
responsive to external stimuli, exogenous cytokinins readily break tuber
dormancy. Many forms of naturally occurring cytokinins are active, includ-
ing free-base, nucleoside, and nucleotide forms of isopentenyl-, trans-, and
cis-zeatins (Suttle, 1998a; Suttle and Banowetz, 2000). Synthetic and meta-
bolically stable cytokinin derivatives are especially effective in breaking
tuber dormancy (Suttle, 2008). Although dormancy is effectively termi-
nated by cytokinin treatment, the resultant sprouts often remain quite
small and fail to elongate. However, simultaneous treatment with cytoki-
nin and gibberellin typically results in sustained sprout growth that more
closely resembles the situation following the natural dormancy break
(Hartmann et al., 2011).

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Complementing these studies are the results from genetic m­ anipulation

of endogenous cytokinin content. Transformation of potatoes with a cytoki-
nin biosynthetic gene from A. tumefaciens resulted in tubers with greatly
elevated levels of endogenous cytokinins and exhibiting precocious sprout-
ing (Ooms and Lenton, 1985). Conversely, overexpression of a cytokinin
oxidase gene, which would be expected to lower cytokinin content, results
in tubers with an extended dormancy (Hartmann et al., 2011). Collectively,
these results are consistent with an essential role for endogenous cytokinins
in the resumption of tuber bud growth following the cessation of dormancy.

14.5.4 Ethylene
As is the case with other vegetative storage organs, potato tubers pro-
duce low (but detectable) amounts of ethylene during storage (Okazawa,
1974). The rate of ethylene production increases coincident with the onset
of sprouting either during the natural dormancy break or following treat-
ment with artificial dormancy-terminating agents (Okazawa, 1974; Suttle,
2009). Exogenous ethylene has a dual effect on tuber dormancy. Short-term
application of high concentrations of ethylene has been reported to hasten
subsequent tuber sprouting, while long-term or continuous ethylene treat-
ment suppresses sprout growth (Rylski et al., 1974). The latter effect was
the impetus for current efforts to commercialize the use of ethylene as a
sprout inhibitor in potato storages (Prange et al., 1998).
Despite these observations, no definitive role for endogenous eth-
ylene in tuber dormancy control was envisaged. This uncertainty was
addressed using an in vitro microtuber system and specific inhibitors of
ethylene action. In this system, ethylene production was highest during
the initial period of tuber development, and chemical inhibition of ethylene
action during the first week of tuber growth resulted in precocious sprout-
ing (Suttle, 1998b). These results suggested that endogenous ethylene
plays an essential role in the induction of microtuber dormancy.
It is surprising that genetic manipulation of ethylene synthesis or
action in potato tubers by sense or antisense approaches has seldom been
reported. Tubers developing on Russet Burbank plants transformed with
either sense or antisense constructs of the ethylene receptor ETR1 dis-
played a number of phenotypic alterations (Haines et al., 2003). When stored
at 20°C, no effects on dormancy duration were noted, but antisense tubers
stored at 4°C failed to sprout after 2 years. Whether this represents a direct
effect of decreased ETR1 expression on tuber dormancy progression or is
a reflection of impaired cold tolerance was not established. Transformation
of potato with antisense constructs of the polyamine biosynthesis gene
SAMDC resulted in increased ethylene evolution and altered tuber pheno-
type, but had no obvious effects on tuber dormancy (Kumar et al., 1996).

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Collectively, these data suggest that endogenous ethylene plays an

essential role in the induction of tuber dormancy, but its participation in
dormancy progression remains to be determined.

14.5.5 Gibberellins
The endogenous gibberellins (GAs) of potato are members of the early
13-hydroxylation pathway, ultimately yielding the bioactive hormone GA 1
(Jones et al., 1988; Xu et al., 1998a). Gibberellins are potent inhibitors of tuber-
ization, and the initiation of tuber formation is preceded by a marked decline
in endogenous GA 1 content in subapical regions of the stolon (Xu et al., 1998a
As the processes of tuber initiation and dormancy inception are thought to
occur together, a decline in GA content may play a role in the initiation of
tuber bud dormancy. As a result of the antagonism between GA content and
tuber formation, the endogenous contents of all GAs are very low in tuber tis-
sues throughout growth, harvest, and early storage (Jones et al., 1988). The
effects of postharvest storage and dormancy progression on endogenous GA
levels in tuber apical bud tissues were determined using GC-MS with internal
standards (Suttle, 2004). The endogenous contents of GA 19, GA 20, and GA 1
declined slightly during early storage when tubers were deeply dormant,
remained at or below harvest levels as dormancy weakened and sprouting
commenced, and increased as sprout growth became more vigorous.
The ability of exogenous GA to prematurely terminate tuber dormancy
was first reported by Brian et al. (1955) and subsequently verified by oth-
ers (Hemberg, 1985). The dormancy-breaking efficacy of individual GAs in
the latter steps of the biosynthetic pathway was GA 19 < GA 20 < GA 1 (Suttle,
2004). As mentioned above, robust sprout growth typically requires the
simultaneous application of both GAs and cytokinin (Hartmann et al., 2011).
The effects of chemical and genetic manipulation of endogenous GA
content have also been reported (Suttle, 2004). Treatment of developing
microtubers with known inhibitors of GA biosynthesis did not extend the
dormant period. No differences in dormancy progression were observed in
studies comparing a wild-type and GA-deficient dwarf mutant of Solanum
andigena. Taken together, these studies suggest that endogenous GAs do
not directly participate in the regulation of tuber dormancy progression,
but may play a critical role in sprout growth following dormancy exit.

14.5.6  Summary and Conclusions

From the foregoing, it is clear that endogenous plant hormones participate
in all phases of tuber dormancy progression, from inception to termination.
Detailed knowledge of the roles in individual hormones during dormancy

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 P OTATO TUBER D ORMAN C Y

has expanded greatly and continues to grow. Nevertheless, there are many
substantive questions that remain unresolved. First and foremost is the
need to acquire detailed quantitative data on the effects of dormancy status
on the hormonal content of tuber buds free from surrounding nongrowing
tissues. Second is the need to develop an understanding of the biochemi-
cal and molecular processes regulating tuber bud hormone biosynthesis
and metabolism during dormancy progression. Last, occurring coinciden-
tally with changes in hormone content are changes in hormone perception
and action. These processes are likely key to developing a more complete
understanding of the physiology of tuber dormancy control and have yet to
receive experimental scrutiny.

14.6  Transcriptional Regulation during Dormancy

14.6.1  Initiation of Tuber Dormancy

As mentioned above, the initiation of dormancy begins with the onset of
tuberization, but there is little evidence linking the expression of specific
genes and the establishment of dormancy in potato meristems. Although
ABA content is positively associated with the induction of tuber dormancy,
the induction of dormancy-specific gene products by ABA has not clearly
been established. It can be concluded that currently there is a lack of evi-
dence linking specific gene expression to the initiation of endodormancy in
potato tubers.
In other perennial systems, such as peach, poplar, and leafy spurge,
short days (SDs) induce bud arrest and entry into dormancy, and these
species exhibit the expression of a MADS-box-like transcription factor dur-
ing the endodormant state (Bielenberg et al., 2008; Hoenicka et al., 2008;
Horvath et al., 2010). Recent evidence using RNA-seq demonstrated that
transcripts for a MADS-box transcription factor are increased in dormant
potato meristems (Campbell et al., submitted), but it is difficult to draw
parallels between perennial systems that are intricately linked to photope-
riod response and potato tubers. The increased expression of a dormancy-
associated MADS-box transcription factor in potato meristems may be
linked to a short photoperiod, but mature tubers are subterranean, and
light-regulated responses terminating MADS-box expression are likely
not significant in the cessation of potato tuber dormancy. The initiation of
tuberization via photoperiod is regulated by a graft-transmissible mecha-
nism, which functions through repression of a CONSTANS-like protein,
which regulates a Flowering Locus T (FT) homolog (Gonzalez-Schain
et  al., 2012). An SD-induced FT homolog has been associated with tuber
formation (Navarro et al., 2011), and FT transcripts are elevated in tuber
meristems during dormancy initiation (Campbell et al., 2008). In poplar

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

SDs, signaling via CONSTANS and FT is linked to seasonally induced

growth arrest and may be directly related to the onset of endodormancy
(Böhlenius et al., 2006). Thus, similarities exist between perennials and
potato in regard to dormancy initiation, and one of the commonalities
seems to be the CONSTANS/FT signaling system.

14.6.2  Dormancy Termination

Transcriptional changes during dormancy progression have been investi-
gated in potato (Campbell et al., 2008), leafy spurge (Horvath et al., 2008),
Japanese pear (Bai et al., 2013), chestnut (Santamaria et al., 2011), poplar
(Ko et al., 2011), apricot (Yamane et al., 2008), and peach (Leida et al.,
2010). Alterations in transcript abundances for proteins that regulate sym-
plastic isolation of meristems during dormancy have been demonstrated in
woody perennials (Rinne et al., 2001, 2011) and are discussed in a previous
review (Sonnewald and Sonnewald, 2014).
A number of studies have examined genetic or transcriptional changes
following termination of bud dormancy using chemicals such as hydrogen
cyanamide (HC) on grapes (Or et al., 2002) and bromoethane on potatoes
(Campbell et al., 2008; Destefano-Beltrán et al., 2006a). In potatoes, deoxy-
uridinetriphosphatase (dUTPase) has been shown to be an early indica-
tor of natural dormancy cessation (Senning et al., 2010). Analysis using
microarrays revealed a decrease in transcripts that are inducible by ABA
as dormancy cessation progressed (Campbell et al., 2008). This same study
revealed that chemical release of potato dormancy using bromoethane
(BE) and natural dormancy cessation increased the expression of genes
encoding for oxoglutarate-dependent oxygenase.
Using flow cytometry and labeling of DNA, it has been demonstrated
that nuclei isolated from dormant potato meristems are arrested in the G1
position of the cell cycle (Campbell et al., 1996). Transcript analysis using
RNA-seq shows an increase in genes associated with DNA replication, as
well as transcripts encoding proteins involved with the progression of cells
from a G1 block (Campbell et al., submitted). In this same study, transcripts
encoding for histone proteins, cyclins, and cell division regulation were
induced by the cytokinin agonist 1-(α-ethylbenzyl)-3-nitroguanidine (NG).
This induction was rapid and occurred within 4 days of treating dormant
tubers with NG. These results indicate that the dormant state results in the
inability of tissues to respond to cytokinins.
As dormancy ends, there is decreased expression of genes encoding
WRKY transcription factors, which are regulated by ABA and tissue dehy-
dration (Rushton et al., 2010). Campbell et al. (2014) also demonstrated
that expression of transcripts encoding gibberellin 20-oxidase (an enzyme
involved with GA biosynthesis) increases with dormancy cessation.

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 P OTATO TUBER D ORMAN C Y

Additional transcripts involved with DNA replication and cell replication,

such as histones and cyclins, are increased with termination of dormancy,
as are transcripts for type A response proteins, which have been shown to
be induced by cytokinins (To et al., 2007). These results suggest that the
transition from dormant to nondormant involves the decrease in expression
of ABA-regulated genes, an increase in genes regulated by cytokinin and
involved with cell division, and an increase in transcripts associated with
GA biosynthesis.

14.6.3  Epigenetic Control of Tuber Dormancy

Structural changes to chromatin (i.e., epigenetic regulation) are correlated
with dormancy status in a number of perennial species. Chromatin restruc-
turing has been associated with dormancy in leafy spurge (Horvath et al.,
2006), poplar (Conde et al., 2013; Druart et al., 2007), chestnut (Santamaria
et al., 2009), peach (Leida et al., 2010), and potato (Law and Suttle, 2003). In
potato, de-methylation of CG islands is associated with endodormancy ter-
mination (Law and Suttle, 2003). Multiacetylation of histone H3.1/3.2 and a
transient acetylation of histone H4 are found in potato tubers following natural
and chemically induced cessation of endodormancy (Law and Suttle, 2004).

14.7  Control of Sprouting in Storage

14.7.1  Introduction and Importance

Controlling sprout development is an important management criterion
for all stored potatoes. If proper sprout control is not maintained, signifi-
cant and detrimental impacts will occur to tuber quality, and the ability to
store for extended periods of time is diminished. Managing sprout devel-
opment allows for a yearlong supply of potatoes and maintenance of qual-
ity throughout the cold chain and marketing. Sprouting causes increased
weight loss, due to both respiration and transpiration losses, and physically
impedes air movement through the bulk potato pile or box. Temperature
in a potato storage is modulated by the ability to supply cooler air to the
potatoes. Physical obstruction lessens the ability to manage the storage
due to increased pile temperatures, and thus may increase the potential
for disease problems to develop. Initiation of sprout development coincides
with physiological changes and can be noticeable in carbohydrate changes
within the tuber. Starch-to-sugar conversions can be detected on a gross
scale. This can have considerable impact on the processing quality of pota-
toes. Visible sprouts on fresh pack or ware potatoes are not acceptable to
consumers. Minimizing sprout development of seed potatoes is important

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relative to physiological aging and the potential to knock off the sprouts
during handling. The latter could cause an increase in stem numbers per
eye and may be an entry point for pathogen invasion.
The current common means to control or minimize sprouting include
cultivar selection, storage temperature, and chemical product application.

14.7.2  Dormancy, Cultivar Selection, and

Storage Temperature
Potato cultivars differ in dormancy length and sprouting behavior (Bogucki
and Nelson, 1980). As described above (Section 14.2), after harvest and
during storage, potatoes pass through three stages of dormancy: endodor-
mancy, para­dormancy, and ecodormancy. Regardless of the type of dor-
mancy displayed by a potato, the development of sprout meristematic tissue
is considered the end of dormancy, also referred to as dormancy break. In
the commercial potato industry, it is necessary to control sprouting beyond
the time when inherent dormancy has ended to ensure a yearlong supply of
potatoes to the market.
Endodormancy varies substantially between cultivars and storage
temperature. Exposure to lower storage temperatures can retard sprout
growth (Gichohi and Pritchard, 1995; Mehta and Kaul, 1991; Tabori and
Hayashi, 1999), although that strategy is limited by the end use of the potato.
Depending on cultivar, low storage temperatures cause increased starch-to-
sugar conversions (Isherwood, 1976; van der Plas, 1987). This increase in
reducing sugars causes products to fry darker, resulting in unacceptable fry
color for consumers. Therefore, utilizing low-temperature storage for sprout
control can be a limitation for potatoes intended for processing. There is
no permanent loss of sprouting capacity in tubers stored at low tempera-
tures. Upon removal and exposure to handling and warmer temperatures,
the potential for sprout development increases. Low storage temperature
is routinely implemented for fresh market potatoes. This can be effective
in situations when potatoes are consumed relatively soon after packing
and distribution. Long-distance transport to the consumer, in conjunction
with time and suboptimal conditions, may promote sprout development.
Otherwise, another method to suppress sprout development is necessary.
Seed potato storage relies on cold temperature (~3°C) and may employ a
mild sprout suppressant with short-dormancy cultivars, if necessary.

14.7.3  Chemical Control

Sprout control is typically accomplished utilizing a chemical sprout inhibi-
tor and cold storage temperatures (<7°C). The success of sprout control

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 P OTATO TUBER D ORMAN C Y

will be dependent on cultivar, how the crop was grown, storage tempera-
ture, and the rate and timing of the product application. It also will vary
greatly between the type of sprout inhibitor and method of application.
Differences in cultivar dormancy length and sprout development charac-
teristics (Kleinkopf and Olsen, 2003) may warrant different application tim-
ings of a sprout inhibitor.
There are several commercialized products utilized for sprout sup-
pression. Not all listed are registered for use worldwide.

Chlorpropham (CIPC): Aerosol and spray applications

Maleic hydrazide: Applied in the field
Essential oils (e.g., clove oil, mint oils, carvone)
Naphthalenes (1,4-DMN and 2,6-DIPN)
Unsaturated ketone (3-decen-2-one)
Ethylene gas
Stabilized hydrogen peroxide
Irradiation (cobalt-60 and electron beam)

Chlorpropham, isopropyl N-(3-chlorophenyl carbamate) (CIPC), is the

most commonly used and effective sprout inhibitor (Kalt et al., 1999) world-
wide in the potato industry. It is applied as a thermal fog or aerosol in the
storage building, or applied as an aqueous emulsion concentrate spray or
dust formulation as the potatoes are conveyed into storage or after wash-
ing and packing. CIPC inhibits sprout development by interfering with cell
division, and it is necessary to apply it prior to sprout formation (Ravanel
and Tissut, 1984 The most common application of CIPC is as a thermal
aerosol fog introduced into the storage building via the ventilation system
(Conte and Imbroglini, 1995; Kleinkopf et al., 1997; Noel et al., 2004). CIPC
is typically applied after wound healing has occurred. Modifications to
application technology have been made to minimize the risk of combus-
tion by-products, such as ethylene, that may impact sugar accumulation
and negative effects on fry or chip color. CIPC is highly effective even at
low concentrations and is a volatile chemical that is lost or degraded and
redistributed with time inside the storage structure (Corsini et al., 1979).
Multiple applications may be necessary depending on the rate and applica-
tion method. The longevity of sprout control is correlated with the initial
application rate and residue on the tuber (Kim et al., 1972; Boyd et al., 1982;
Kleinkopf et al., 1997). Cultivars can also differ in sensitivity to CIPC and,
therefore, the necessary residue level for sprout suppression. Also, depend-
ing on the end use of the potato, cultivars will be held at various storage
temperatures that can alter the degradation of CIPC on the tuber (Corsini
et al., 1979; Mondy et al., 1992).
There are several other products that have sprout suppression activity
either independently or in combination with other sprout control agents or cold

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storage temperatures. Maleic hydrazide (1,2-dihydropyridazine-3, 6-dione), a

plant growth regulator, can be applied to actively growing potatoes during
the growing season for sprout control later in storage. Cultivars differ in
response to maleic hydrazide, and sprout control in long-term storage var-
ies with the rate and timing of application (Caldiz et al., 2001). In general,
maleic hydrazide delays sprouting by approximately 30 days and retards
sprout elongation for the duration of storage. Previous research has shown
substituted naphthalenes as having short-term sprout suppression prop-
erties (Beveridge et al., 1981; Lewis et al., 1997). Both Beveridge et al.
(1981) and Knowles et al. (2005) reported 1,4-DMN as a weak sprout sup-
pressant that could be used on seed potatoes. Recently, the combination
of naphthalenes and lower rates of CIPC has been introduced to the U.S.
industry (Beaver et al., 2003). The combination of the two products pro-
vided similar sprout control when compared to a higher label rate of CIPC
applied alone. Irradiation utilizing either gamma (cobalt-60) or electron
beam radiation sources can be an effective sprout control tool at relatively
low dose rates (Thomas and Sparks, 1984; Todoriki and Hayashi, 2004;
Frazier et al., 2006; Rezaee et al., 2011). The economics and increased sus-
ceptibility to disease and tuber sugar concentrations may limit the useful-
ness of this process (Burton, 1975; Frazier et al., 2006; Rezaee et al., 2011).
Essential oils, such as mint (spearmint, Mentha spicata) and pep-
permint (Mentha piperata) and clove (Syzygium aromaticum) oils and car-
vone (derived from caraway seed [Carum carvi]), have shown efficacy in
suppressing sprout development in potatoes (Vaughn and Spencer, 1993;
Hartmans et al., 1995; Oosterhaven et al., 1995; Coleman et al., 2001;
Kleinkopf and Olsen, 2003). The application methodology differs from that
of CIPC with these products, and repeated applications are generally nec-
essary since physical meristematic damage is the primary mode of action
with these compounds (Coleman et al., 2001; Baydar and Karadoga, 2004).
These volatile products must be applied after sprout initiation or elongation
has occurred. They are typically applied as a thermal fog, wicking, cold
aerosol, or spray. The application method will depend on the storage design
and chemical active ingredient. The typical length of sprout suppression
ranges from 2 to 5 weeks before reapplication is necessary, dependent on
cultivar and storage temperature. Essential oil treatments are nonpersis-
tent and can be used to suppress the sprouting of seed potatoes (Sorce
et al., 1997).
Stabilized hydrogen peroxide suppresses potato sprouting by physi-
cally damaging the meristematic sprout tissue (Afek et al., 2000). It is typi-
cally applied through the humidification system of a potato storage facility.
Multiple applications are necessary to continuously damage any new sprout
tissue development. A recently registered saturated ketone, 3-decene-
2-one, has provided an additional option for sprout control in the United
States and Canada (Frazier et al., 2008). This sprout inhibitor physically

466
 P OTATO TUBER D ORMAN C Y

damages sprout meristematic tissue and is applied as a thermal aerosol fog

in the storage facility. Sprout suppression by 3-decen-2-one is not p
­ ermanent,
and aerosol applications will give approximately 3–8 weeks of sprout con-
trol, depending on variety and storage temperature.
Ethylene gas has recently been utilized for sprout suppression,
although the plant growth regulator has been shown to both promote and
inhibit sprouting (Elmer, 1936). Tuber response is dependent on duration
and level of exposure (Rylski et al., 1974). Ethylene potato sprout suppres-
sant acts by inhibiting the elongation of the growing sprouts (Prange et al.,
1998). Exposure to ethylene must be continuous to maintain sprout sup-
pression (Daniels-Lake et al., 2005). Ethylene-induced sprout inhibition is
nonpersistent, and sprout development will continue once the exposure is
removed. Ethylene can be used as a sprout suppressant for seed potatoes.

14.8  Conclusions and Future Perspectives

The regulation of tuber dormancy is of both academic and practical impor-
tance. The internal processes that regulate growth arrest in plant meristems
are poorly understood and represent a major gap in our understanding of plant
development. From a more practical view, plant meristem dormancy affects
many aspects of plant husbandry and agriculture. From the germination of
both weed and crop seeds to the arrest and resumption of meristem activity
in perennial plants that is critical to survival in unfavorable environments, a
greater understanding of the internal processes that control dormancy pro-
gression is needed to ensure maximum agricultural productivity in all envi-
ronments where plants are cultivated. Although significant progress has been
made in identifying the cognate factors affecting potato tuber dormancy, there
is still much to learn. Together with ever-more sensitive methods of hormone
and metabolite quantification, the use of selective gene-disrupting technolo-
gies will greatly expand the frontiers of knowledge in this area. Collectively,
these and other advancements may ultimately lead to the identification of the
true master switch that is the Holy Grail of dormancy research.

References
Addicott, F.T., and Carns, H.R. 1983. History and introduction. In Addicott,
F.T. (ed.), Abscisic Acid. Praeger, New York, 1–22.
Afek, U.J., Orenstein, J., and Nuriel, E. 2000. Using HPP (hydrogen peroxide
plus) to inhibit potato sprouting during storage. American Journal of
Potato Research 77, 63–65.
Bai, S., Saito, T. Sakamoto, D., Ito, A., Fujii, H., and Moriguchi, T. 2013.
Transcriptome analysis of Japanese pear (Pyrus pyrifolia Nakai) flower

467
P OST H AR V EST RI P ENING P H Y SIOLOG Y

buds transitioning through endodormancy. Plant and Cell Physiology 54,

1132–1151.
Baydar, H., and Karadogan, T. 2004. The effects of volatile oils on in vitro
potato sprout growth. Potato Research 46, 1–8.
Beaver, R.G., Devoy, M., Schafer, R., and Riggle, B.D. 2003. CIPC and 2,6-
DIPN sprout suppression of stored potatoes. American Journal of Potato
Research 80, 311–316.
Beveridge, J.L., Dalziel, J., and Duncan, H.J. 1981. Dimethylnaphthalene as
a sprout suppressant for seed and ware potatoes. Potato Research 24,
77–88.
Bielenberg, D., Wang, Y., Li, Z., Zhebentyayeva, T., Fan, S., Reighard,
G.L., Scorza, R., and Abbott, A. 2008. Sequencing and annotation of the
evergrowing locus in peach (Prunus persica L.) reveals a cluster of six
MADS-box transcription factors as candidate genes for regulation of ter-
minal bud formation. Tree Genetics and Genomes 4, 495–507.
Bodlaender, K.B.A., Lugt, C., and Marinum, J. 1964. The induction of second
growth in potato tubers. European Potato Journal 7, 57–71.
Bogucki, S., and Nelson, D.C. 1980. Length of dormancy and sprouting char-
acteristics of ten potato cultivars. American Potato Journal 57, 151–157.
Böhlenius, H., Huang, T., Charbonnel-Campa, L., Brunner, A.M., Jansson, S.,
Strauss, S.H., and Nilsson, O. 2006. CO/FT regulatory module controls
timing of flowering and seasonal growth cessation in trees. Science 312,
1040–1043.
Boyd, I., Dalziel, J., and Duncan, H.J. 1982. Studies on potato sprout suppres-
sants. 5. The effect of chlorpropham contamination on the performance
of seed potatoes. Potato Research 18, 109–115.
Brian, P.W., Hemming, H.G., and Radley, M. 1955. A physiological comparison
of gibberellic acid with some auxins. Physiologia Plantarum 8, 899–912.
Burton, W.G. 1963. Concepts and mechanism of dormancy. In Ivins, J.D., and
Milthorpe, F.L. (eds.), The Growth of the Potato. Butterworths, London,
17–41.
Burton, W.G. 1975. The immediate effect of gamma irradiation upon the
sugar content of potatoes previously stored at 2, 4.5, 6, 10, and 15.5°C.
Potato Research 18, 109–115.
Burton, W.G. 1989. The Potato. 3rd ed. Longman Scientific and Technical,
Essex, 470–504.
Burton, W.G., and Meigh, D.F. 1971. The production of growth-suppressing
volatile substances by stored potato tubers. Potato Research 14, 96–101.
Caldiz, D.O., Fernandez, L.V., and Inchausti, M.H. 2001. Maleic hydrazide
effects on tuber yield, sprouting characteristics, and French fry process-
ing quality in various potato (Solanum tuberosum L.) cultivars grown under
Argentinian conditions. American Journal of Potato Research 78, 119–128.
Campbell, M.A., Segear, E., Beers, L., Kanuber, D., and Suttle, J. 2008.
Dormancy in potato tuber meristems: Chemically induced cessation
in dormancy matches the natural process based on transcript profiles.
Functional and Integrative Genomics 8, 317–328.

468
 P OTATO TUBER D ORMAN C Y

Campbell, M.A., Suttle, J., Douches, D.S., and Buell, C.R. 2014. Transcriptome
changes in potato meristems during termination of endodormancy with
1-(α-ethylbenzyl)-3-nitroguanidine. Functional and Integrative Genomics
14, 789–799.
Campbell, M.A., Suttle, J.C., and Sell, T.W. 1996. Changes in cell cycle ­status
and expression of p34cdc2 kinase during potato tuber meristem dor-
mancy. Physiologia Plantarum 98, 743–752.
Coleman, W.K. 2000. Physiological ageing of potato tubers: A review. Annals
of Applied Biology 137, 189–199.
Coleman, W.K., Lonergan, G., and Silk, P. 2001. Potato sprout growth sup-
pression by methone and neomenthol, volatile oil components of
Minthostachys, Satureja, Bystropogon, and Mentha species. American
Journal of Potato Research 78, 345–354.
Conde, D., Gonzalez-Melendi, P., and Allona, I. 2013. Poplar stems show
opposite epigenetic patterns during winter dormancy and vegetative
growth. Trees 27, 311–320.
Conte, E., and Imbroglini, G. 1995. Presence of sprout inhibitor residues in
potatoes in relation to application technologies. Journal of Agricultural
and Food Chemistry 43, 2985–2987.
Corsini, D., Stallknecht, G., and Sparks, W. 1979. Changes in chlorpropham
residues in stored potatoes. American Potato Journal 56, 43–50.
Daniels-Lake, B.J., Prange, R.K., Nowak, J., Asiedu, S.K., and Walsh, J.R. 2005.
Sprout development and processing quality changes in potato tubers
stored under ethylene. 1. Effects of ethylene concentration. American
Journal of Potato Research 82, 389–397.
Destefano-Beltrán, L., Knauber, D., Huckle, L., and Suttle, J.C. 2006a. Effects
of postharvest storage and dormancy status on ABA content, metabo-
lism, and expression of genes involved in ABA biosynthesis and metab-
olism in potato tuber tissues. Plant Molecular Biology 61, 687–697.
Destefano-Beltrán, L., Knauber, D., Huckle, L., and Suttle, J.C. 2006b.
Chemically forced dormancy termination mimics natural dormancy
progression in potato tuber meristems by reducing ABA content and
modifying expression of genes involved in regulating ABA synthesis
and metabolism. Journal of Experimental Botany 57, 2879–2886.
Druart, N., Johansson, A., Baba, A., Schrader, J., Sjodin, A., Bhalerao, R.R.,
Resman, L., Trygg, J., Mortiz, T., and Bhalerao, R.P. 2007. Environmental
and hormonal regulation of the activity-dormancy cycle in the cambial
meristem involves stage-specific modulation of transcriptional and met-
abolic networks. Plant Journal 50, 557–573.
Dun, E.A., Brewer, P.B., and Beveridge, C.A. 2009. Strigolactones: Discovery
of the elusive shoot branching hormone. Trends in Plant Science 14,
364–372.
El-Antably, Wareing, P.F., and Hillman, J. 1967. Some physiological responses
to D,L-abscisin (dormin). Planta 73, 74–90.
Elmer, O.H. 1936. Growth inhibition in the potato caused by a gas emanating
from apples. Journal of Agricultural Research 52, 609–626.

469
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Emilsson, B. 1949. Studies on the rest period and dormant period in the
potato tuber. Acta Agriculturae Sueciae 3, 189–284.
Faivre-Rampant, O., Cardle, L., Marshall, D., Viola, R., and Taylor, M.A. 2004.
Changes in gene expression during meristem activation processes in
Solanum tuberosum with a focus on the regulation of an auxin response
factor gene. Journal of Experimental Botany 55, 613–622.
Frazier, M.J., Kleinkopf, G.E., Brey, R., and Olsen, N. 2006. Potato sprout
inhibition and tuber quality after treatment with high-energy ionizing
radiation. American Journal of Potato Research 82, 29–37.
Frazier, M.J., Olsen, N., Kleinkopf, G.E., and Knowles, N.R. 2008. Potato
sprout suppression potential using C9-ketone. American Journal of
Potato Research 85, 9–15.
Freyre, R., Warnke, S., Sosinski, B., and Douches, D.S. 1994. Quantitative
trait locus analysis of tuber dormancy in diploid potato (Solanum spp.).
Theoretical and Applied Genetics 89, 474–480.
Gichohi, E.G., and Pritchard, M.K. 1995. Storage temperature and maleic
hydrazide effects on sprouting, sugars, and fry color of Shepody pota-
toes. American Potato Journal 72, 737–747.
Gonzalez-Schain, N.D., Diaz-Mendoza, M., Zurczak, M., and Suarez-Lopez, P.
2012. Potato CONSTANS is involved in photoperiodic tuberization in a
graft-transmissible manner. Plant Journal 70, 678–690.
Haines, M.M., Shiel, P.J., Fellman, J.K., and Berger, P.H. 2003. Abnormalities
in growth, development and physiological responses to biotic and abi-
otic stress in potato (Solanum tuberosum) transformed with Arabidopsis
ETR1. Journal of Agricultural Science 141, 333–347.
Hartmann, A., Senning, M., Heddon, P., Sonnewald, U., and Sonnewald, S.
2011. Reactivation of meristem activity and sprout growth in potato
tubers requires both cytokinin and gibberellin. Plant Physiology 155,
776–796.
Hartmans, K.J., Diepenhorst, P., Bakker, W., and Gorris, L.G.M. 1995. The
use of carvone in agriculture: Sprout suppression of potatoes and anti-
fungal activity against potato tuber and other diseases. Industrial Crops
and Products 4, 3–13.
Hemberg, T. 1970. The action of some cytokinins on the rest-period and
the content of acid growth-inhibiting substances in potato. Physiologia
Plantarum 5, 115–129.
Hemberg, T. 1985. Potato rest. In Li, P.H. (ed.), Potato Physiology. Academic
Press, New York, 353–388.
Hiller, L.K., Koller, D.C., and Thornton, R.E. 1985. Physiological disorders of
potato tubers. In Li, P. (ed.), Potato Physiology. Academic Press, Orlando,
FL, 389–456.
Hoenicka, H., Nowitzki, O., Hanelt, D., and Fladung, M. 2008. Heterologous
over-expression of the birch FRUITFUL-like MADS-box gene
BpMADS4 prevents normal senescence and winter dormancy in
Populus tremula L. Planta 227, 1001–1011.

470
 P OTATO TUBER D ORMAN C Y

Horvath, D., Anderson, J.V., Soto-Suarez, M., and Chao, W.S. 2006.
Transcriptome analysis of leafy spurge (Euphorbia esula) crown buds dur-
ing shifts in well-defined phases of dormancy. Weed Science 54, 821–827.
Horvath, D., Chao, W., Suttle, J.C., Thimmapuram, J., and Anderson, J.V.
2008. Transcriptome analysis identifies novel responses and potential
regulatory genes involved in seasonal dormancy transitions of leafy
spurge (Euphorbia esula L.). BMC Genomics 9, 536.
Horvath, D., Sung, S., Kim, D., Chao, W., and Anderson, J. 2010. Char-
acterization, expression, and function of DORMANCY ASSO­CIATED
MADS BOX genes from leafy spurge. Plant Molecular Biology 73, 169–179.
Isherwood, F.A. 1976. Mechanism of starch-sugar interconversion in Solanum
tuberosum. Phytochemistry 15, 33–41.
Jones, M.G., Horgan, R., and Hall, M.A. 1988. Endogenous gibberellins in the
potato (Solanum tuberosum). Phytochemistry 27, 7–10.
Kalt, W., Prange, R., and Daniels-Lake, B. 1999. Alternative compounds for
the maintenance of processing quality of stored potatoes during stor-
age. Journal of Food Processing and Preservation 23, 71–81.
Kim, M.S.L., Ewing, E.E., and Sieczka, J.B. 1972. Effects of chlorpropham
(CIPC) on sprouting of individual potato eyes and on plant emergence.
American Potato Journal 49, 420–431.
Klee, H., Horsch, R., and Roger, S. 1987. Agrobacterium-mediated plant trans-
formation and its further applications to plant biology. Annual Review of
Plant Physiology 38, 467–486.
Kleinkopf, G.E., Brandt, T.L., Frazier, M.J., and Moller, G. 1997. CIPC resi-
dues on stored Russet Burbank potatoes. 1. Maximum label application.
American Potato Journal 74, 107–117.
Kleinkopf, G.E., and Olsen, N. 2003. Storage management. In Stark, J.,
and Love, S. (eds.), Potato Production Systems. University of Idaho
Agricultural Communications, Moscow, 363–381.
Knowles, N.R., Knowles, L., and Haines, M.M. 2005. 1,4-Dimethylnaphthalene
treatment of seed potatoes affects tuber size distribution. American
Journal of Potato Research 82, 179–190.
Ko, J.H., Prassinos, C., Keathley, D., and Han, K.H. 2011. Novel aspects of
transcriptional regulation in the winter survival and maintenance mech-
anism of poplar. Tree Physiology 31, 208–225.
Kloosterman, B., Navarro, C., Bijsterbosch, G., Lange, T., Prat, S., Visser,
R.G.F., and Bachem, C.W.B. 2007. StGA20ox1 is induced prior to stolon
swelling and controls GA levels during potato tuber development. Plant
Journal 52, 362–373.
Korasick, D.A., Enders, T.A., and Strader, L.C. 2013. Auxin biosynthesis and
storage forms. Journal of Experimental Botany 64, 2541–2555.
Kumar, A., Taylor, M.A., Mad Arif, S.A., and Davies, H.V. 1996. Potato plants
expressing antisense and sense S-adenosylmethionine decarboxylase
(SAMDC) trans genes show altered levels of polyamines and ethylene:
antisense plants display abnormal phenotypes. Plant Journal 9, 147–158.

471
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Kyozuka, J. 2007. Control of shoot and root meristem function by cytokinin.

Current Opinion in Plant Biology 10, 442–446.
Lang, G.A., Early, J.D., Martin, G.C., and Darnell, R.L. 1987. Endo-,
para-,  and eco-dormancy: Physiological terminology and classification
for dormancy research. HortScience 22, 371–377.
Law, R.D., and Suttle, J.C. 2003. Transient decreases in methylation at
5-CCGG-3 sequences in potato (Solanum tuberosum L.) meristem DNA
during progression of tubers through dormancy precede the resump-
tion of sprout growth. Plant Molecular Biology 51, 437–447.
Law, R.D., and Suttle, J.C. 2004. Changes in histone H3 and H4 multi-­
acetylation during natural and forced dormancy break in potato tubers.
Physiologia Plantarum 120, 642–649.
Leida, C., Terol, J., Marti, G., Agusti, M., Llacer, G., Badenes, M.L., and Rios,
G. 2010. Identification of genes associated with bud dormancy release in
Prunus persica by suppression subtractive hybridization. Tree Physiology
30, 655–666.
Lewis, M.D., Kleinkopf, G.E., and Shetty, K.K. 1997. Dimethylnaphthalene
and diisopropylnaphthalene for potato sprout control in storage. 1.
Application methodology and efficacy. American Journal of Potato
Research 74, 183–197.
Liu, X., Zhang, H., Zhao, Y., Feng, Z., Li, Q., Yang, H.Q., Luan, S., Li, J., and
He, Z.H. 2013. Auxin control seed dormancy through stimulation of
abscisic acid signaling by inducing ARF-mediated ABI3 activation in
Arabidopsis. Proceedings of the National Academy of Sciences USA 110,
15485–15490.
Mehta, A., and Kaul, H.N. 1991. Effect of sprout inhibitors on potato tubers
(Solanum tuberosum L.) stored at ambient or reduced temperatures.
Potato Research 34, 443–450.
Meigh, D.F., Authur, A., Filmer, E., and Self, R. 1973. Growth-inhibiting
volatile aromatic compounds produced by Solanum tuberosum L.
Phytochemistry 12, 987–993.
Mok, D.W.S., and Mok, M.C. 2001. Cytokinin metabolism and action. Annual
Review of Plant Physiology and Plant Molecular Biology 52, 89–118.
Mondy, N.I., Munshi, C.B., and Seetharaman, K. 1992. Residue levels of iso-
propyl-N-(3-chlorophenyl)-carbamate (CIPC) in potatoes as affected by
level of application, storage time, temperature, and method of cooking.
Food Research International 25, 375–379.
Navarro, C., Abelenda, J.A., Cruz-Oró, E., Cuellar, C.A., Tamaki, S., Silva,
J., Shimamoto, K., and Prat, S. 2011. Control of flowering and storage
organ formation in potato by Flowering Locus T. Nature 478, 119–123.
Ohmiya, A. 2009. Carotenoid cleavage dioxygenases and their apocarotenoid
products in plants. Plant Biotechnology 26, 351–358.
Okazawa, Y. 1974. A relation between ethylene evolution and sprouting of
potato tuber. Journal of Faculty of Agriculture of Hokkaido University 57,
43–45.

472
 P OTATO TUBER D ORMAN C Y

Ooms, G., and Lenton, J.R. 1985. T-DNA genes to study plant development:
Precocious tuberization and enhanced cytokinins in A. tumefaciens
transformed potato. Plant Molecular Biology 5, 205–212.
Oosterhaven, K., Hartmans, K.J., and Scheffer, J.C. 1995. Inhibition of potato
sprout growth by carvone enantiomers and their bioconversion in
sprouts. Potato Research 38, 219–230.
Or, E., Vilozny, I., Fennel, A., Eyal, Y., and Ogrodovitch, A. 2002. Dormancy
in grape buds: Isolation and characterization of catalase cDNA and anal-
ysis of its expression following chemical induction of bud dormancy
release. Plant Science 162, 121–130.
Pasare, S.A., Ducreux, L.J.M., Morrie, W.L., Campbell, R., Sharma, S.K.,
Roumeliotis, E., Kohlen, W., van der Krol, S., Bramley, P.M., Roberts,
A.G., Fraser, P.D., and Taylor, M.A. 2013. The role of the potato
(Solanum tuberosum) CCD8 gene in stolon and tuber development.
New Phytology 198, 1108–1120.
Prange, R.K., Kalt, W., Daniels-Lake, B.J., Liew, C.L., and Page, R.T. 1998.
Using ethylene as a sprout control agent in stored ‘Russet Burbank’
potatoes. Journal of the American Society for Horticultural Science 123,
463–469.
Quarrie, S.A. 1982. Droopy: A wilty mutant of potato deficient in abscisic acid.
Plant Cell and Environment 5, 23–26.
Rezaee, M., Almassi, M., Farahani, A.M., Minaei, S., and Khodadadi, M.
2011. Potato sprout inhibition and tuber quality after postharvest treat-
ment with gamma irradiation on different dates. Journal of Agricultural
Science and Technology 13, 829–841.
Rinne P.L.H., Kaikuranta, P.M., and van der Schoot, C. 2001. The shoot apical
meristem restores its symplastic organization during chilling-induced
release from dormancy. Plant Journal 26, 249–264.
Rinne, P.L.H., Welling, A., Vahala, J., Ripel, L., Ruonala, R., Kangasjarvi, J., and van
der Schoot, C. 2011. Chilling of dormant buds hyperinduces FLOWRING
LOCUS T and recruits GA-inducible 1.3-β-glucanases to reopen signal
conduits and release dormancy in Populus. Plant Cell 23, 130–146.
Rushton, P.J., Somssich, I.E., Ringler, P., and Shen, Q.J. 2010. WRKY tran-
scription factors. Trends in Plant Science 15, 247–258.
Rylski, I., Rappaport, L., and Pratt, H.K. 1974. Dual effects of ethylene on
potato dormancy and sprout growth. Plant Physiology 131, 419–429.
Sakakibara, H. 2006. Cytokinins: activity, biosynthesis, and translocation.
Annual Review of Plant Biology 57, 431–439.
Samish, R.M. 1954. Dormancy in woody plants. Annual Review of Plant
Physiology 5, 183–204.
Santamaria, M.E., Hasbun, R., Valera, M.J., Meijon, M., Valledor, L.,
Rodriguez, J.L., Toorop, P., Canal, M.J., and Rodriguez, R. 2009.
Acetylated H4 histone and genomic DNA methylation patterns during
bud set and bud burst in Castanea sativa. Journal of Plant Physiology
166, 1360–1369.

473
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Santamaria, M.E., Rodriguez, R., Canal, M.J., and Toorop, P.E. 2011.
Transcriptome analysis of chestnut (Castanea sativa) tree buds sug-
gests a putative role for epigenetic control of bud dormancy. Annals of
Botany 108, 485–498.
Schippers, P.A. 1956. De duur van de rustperiode van een veertigtal aardap-
pelrassen, bewaard bijvershillende constante temperaturen. Publ.
Aardappelbew. Wageningen, Ser. A (112), 10.
Schmülling, T., Fladung, M., Grossmann, K., and Schell, J. 1993. Hormonal
content and sensitivity of transgenic tobacco and potato plants express-
ing single rol genes of Agrobacterium rhizogenes T-DNA. Plant Journal
3, 371–382.
Senning, M., Sonnewald, U., and Sonnewald, S. 2010. Deoxyuridinetriphos­
phatase expression defines the transition from dormant to sprouting
potato tuber buds. Molecular Breeding 26, 525–531.
Šimko, I., McMurry, S., Yang, H.M., Manschot, A., Davies, P.J., and Ewing,
E.E. 1997. Evidence from polygene mapping for a causal relation-
ship between potato tuber dormancy and abscisic acid content. Plant
Physiology 115, 1453–1459.
Simmonds, N.W. 1964. The genetics of seed and tuber dormancy in the culti-
vated potatoes. Heredity 19, 489–504.
Sonnewald, S., and Sonnewald, U. 2014. Regulation of potato tuber sprouting.
Planta 239, 27–38.
Sorce, C., Lombardi, L., Giorgetti, L., Parisi, B., Ranalli, P., and Lorenzi, R.
2009. Indoleacetic acid concentration and metabolism changes during
bud development in tubers of two potato (Solanum tuberosum) cultivars.
Journal of Plant Physiology 166, 1023–1033.
Sorce, C., Lorenzi, R. Ceccarelli, N., and Ranalli, P. 2000. Changes in free
and conjugated IAA during dormancy and sprouting of potato tubers.
Australian Journal of Plant Physiology 27, 371–377.
Sorce, C., Piaggesi, A., Ceccarelli, N., and Lorenzi, R. 1996. Role and metabo-
lism of abscisic acid in potato tuber dormancy and sprouting. Journal of
Plant Physiology 149, 548–552.
Sorce, C.R., Lorenzi, R., and Ranalli, P. 1997. The effects of (S)-carvone on
seed potato tuber dormancy and sprouting. Potato Research 40, 155–161.
Sukhova, L.S., Machackova, I., Eder, J., Bibik, N.D., and Korableva, N.P. 1993.
Changes in the levels of free IAA and cytokinins in potato tubers during
dormancy and sprouting. Biologia Plantarum 35, 387–391.
Suttle, J.C. 2008. Effects of synthetic phenylurea and nitroguanidine cytoki-
nins on dormancy break and sprout growth in Russet Burbank minitu-
bers. American Journal of Potato Research 85, 121–128.
Suttle, J.C. 1998a. Postharvest changes in endogenous cytokinins and
cytokinin efficacy in potato tubers in relation to bud endodormancy.
Physiologia Plantarum 103, 59–69.
Suttle, J.C. 1998b. Involvement of ethylene in potato microtuber dormancy.
Plant Physiology 118, 843–848.

474
 P OTATO TUBER D ORMAN C Y

Suttle, J.C. 2002. Dormancy-related changes in cytokinin efficacy and

metabolism in potato tubers during postharvest storage. Plant Growth
Regulation 35, 199–206.
Suttle, J.C. 2003. Auxin-induced sprout growth inhibition: Role of endogenous
ethylene. American Journal of Potato Research 80, 303–309.
Suttle, J.C. 2004. Involvement of endogenous gibberellins in potato tuber dor-
mancy and early sprout growth: A critical evaluation. Journal of Plant
Physiology 161, 157–164.
Suttle, J.C. 2007. Dormancy and sprouting. In Vreugdenhil, D. (ed.),
Potato Biology and Biotechnology: Advances and Perspectives. Elsevier,
Amsterdam, 287–309.
Suttle, J.C. 2008. Effects of synthetic phenylurea and nitroguanidine cytoki-
nins on dormancy break and sprout growth in Russet Burbank minitu-
bers. American Journal of Potato Research 85, 121–128.
Suttle, J.C. 2009. Ethylene is not involved in hormone- and bromoethane-
induced dormancy break in Russet Burbank minitubers. American
Journal of Potato Research 86, 278–285.
Suttle, J.C., and Banowetz, G.M. 2000. Changes in cis-zeatin and cis-zeatin
riboside levels and biological activity during potato tuber dormancy.
Physiologia Plantarum 109, 68–74.
Suttle, J.C., Huckle, L.L., Lu, S., and Knauber, D.C. 2014. Potato tuber cyto-
kinin oxidase/dehydrogenase genes: biochemical properties, activity,
and expression during tuber dormancy progression. Journal of Plant
Physiology 171, 448–457.
Suttle, J.C., and Hultstrand, J.F. 1994. Role of endogenous abscisic acid in
potato microtuber dormancy. Plant Physiology 105, 891–896.
Suttle, J.C., Abrams, S.R., De Stefano-Beltrán, L., and Huckle, L.L. 2012.
Chemical inhibition of potato ABA-8-hydroxylase activity alters in vitro
and in vivo ABA metabolism and endogenous ABA levels but does not
affect potato microtuber dormancy duration. Journal of Experimental
Botany 63, 5717–5725.
Tabori, S., and Hayashi, T. 1999. Some sprouting characteristics of microtu-
bers. Potato Research 40, 155–161.
Thomas, P., and Sparks, W.C. 1984. Radiation preservation of foods of plant
origin. Part 1. Potatoes and other crops. Critical Reviews of Food Science
and Nutrition 19, 327–379.
To, J.P.C., Deruère, J., Maxwell, B.B., Morris, V.F., Hutchison, C.E., Ferreira,
F.J., Schaller, G.E., and Kieber, J.J. 2007. Cytokinin regulates type-A
Arabidopsis response regulator activity and protein stability via two-
component phosphorelay. Plant Cell 19, 3901–3914.
Todoriki, S., and Hayashi, T. 2004. Sprout inhibition of potatoes with soft-
electron (low-energy electron beams). Journal of the Science of Food
Agriculture 84, 2010–2014.
USDA/NASS (2013) Crop Production 2012 Summary. January, 2013.

475
P OST H AR V EST RI P ENING P H Y SIOLOG Y

van den Berg, J.H., Ewing, E.E., Plaisted, R.L., McMurry, S., and Bonierbale,
M.W. 1996. QTL analysis of potato tuber dormancy. Theoretical and
Applied Genetics 93, 317–324.
van der Plas, L.H.W. 1987. Potato tuber storage: Biochemical and physiologi-
cal changes. In Bajaj, Y. (ed.), Biotechnology in Agriculture and Forestry.
Vol. 3. Springer-Verlag, Berlin, 109–135.
Vaughn, S.F., and Spencer, G.F. 1993. Naturally occurring aromatic com-
pounds inhibit potato tuber sprouting. American Potato Journal 70,
527–533.
Xu, X., van Lammeren, A.A.M., Vermeer, E., and Vreugdenhil, D. 1998a. The
role of gibberellin, abscisic acid, and sucrose in the regulation of potato
tuber formation in vitro. Plant Physiology 117, 575–584.
Xu, X., Vreugdenhil, D., and van Lammeren, A.A.M. 1998b. Cell division and
cell enlargement during potato tuber formation. Journal of Experimental
Botany 49, 573–582.
Yamane, H., Kashiwa, Y., Ooka, T., Tao, R., and Yonemori, K. 2008. Suppression
subtractive hybridization and differential screening reveals endodor-
mancy-associated expression of an SVP/AGL24-type MADS-box gene
in lateral vegetative buds of Japanese apricot. Journal of the American
Society for Horticultural Science 133, 707–716.

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Chapter 15

Calcium Deficiency
Disorders in Plants
Sergio Tonetto de Freitas,1 Cassandro Vidal
Talamini do Amarante,2 and Elizabeth J. Mitcham3
1Brazilian Agricultural Research Corporation,
Embrapa Tropical Semi-arid, Petrolina, Pernambuco, Brazil
2Santa Catarina State University, Lages, Santa Catarina, Brazil
3 University of California, Davis, California

Abstract 478
15.1 History of Ca2+ Deficiency Disorders 479
15.2 Role of Ca2+ as an Essential Plant Macronutrient 479
15.3 Symptoms of Ca2+ Deficiency Disorders in Fruit 481
15.3.1 Apple 481
15.3.2 Tomato 484
15.3.3 Watermelon 484
15.3.4 Pepper 484
15.4 Symptoms of Ca2+ Deficiency Disorders in Leafy
Vegetables 485
15.4.1 Lettuce 485
15.4.2 Cauliflower 485
15.4.3 Artichoke 486
15.4.4 Celery 487

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15.5 Potential Mechanisms Regulating Ca2+ Deficiency

Disorders 488
15.5.1  Total Tissue Ca2+ Content 488
15.5.2 Cellular Regulation of Ca2+ Partitioning and
Distribution 490
15.5.3  Other Nutrients 492
15.5.3.1 Nitrogen 492
15.5.3.2  Potassium and Magnesium 493
15.5.3.3 Boron 494
15.5.3.4 Phosphorus 494
15.5.4  Reactive Oxygen Species 495
15.5.5  Growth Regulators 495
15.5.5.1 Growth Regulators Affecting Total
Tissue Ca2+ Content 496
15.5.5.2 Growth Regulators Influencing
Cellular Ca2+ Distribution 498
15.5.5.3 Growth Regulator Effect on
Oxidative Metabolism 499
15.6  Possible Control Strategies 500
15.6.1  At the Tissue Level 500
15.6.2  At the Cellular Level 501
15.7  Final Considerations and Future Research Needs 501
References 502

Abstract
Physiological disorders, such as bitter pit in apple; blossom-end rot in
tomato, watermelon, and pepper; tipburn in lettuce, cauliflower, artichoke;
and blackheart in celery are believed to be triggered by Ca 2+ deficiency and
can strongly reduce crop quality and yield. These disorders are character-
ized by dark brown lesions on distal young and fast-growing tissue. In leafy
vegetables, stunted growth and curling are also common symptoms. The
conserved symptoms and factors leading to Ca 2+ deficiency disorders sug-
gest the existence of conserved mechanisms regulating these disorders in
fruits and vegetables. Suggested mechanisms triggering these disorders
are involved in the inhibition of Ca 2+ accumulation or abnormal regulation of
cellular Ca 2+ partitioning in affected tissues. Interactions between Ca 2+ and
other nutrients in affected tissue have also been suggested to be involved.
Although recent ideas have suggested that oxidative stress may play an
important role in Ca 2+ deficiency disorder development, they remain to be
experimentally analyzed. Considering the complexity of Ca 2+ deficiency

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 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

disorders, control strategies should first identify the genetic and environ-
mental factors triggering mechanisms leading to symptom ­development.
Based on the factors involved, specific approaches can be identified to
effectively inhibit Ca 2+ deficiency disorder development.

15.1  History of Ca2+ Deficiency Disorders

The symptoms of Ca 2+ deficiency disorders in plants have been reported in
the literature for more than a hundred years for different crop species (Jager,
1869; Galloway, 1888; Cobb, 1895; Brooks, 1914). Earlier named only as
physiological disorders due to unknown causes, these disorders were first
suggested to be the result of pathogen infection, toxicity, and plant stress con-
ditions (Smith, 1926; Wedgworth et al., 1926; Chamberlain, 1933; Atanasoff,
1934; Carne and Martin, 1934). Later studies focusing on plant nutrient
requirements revealed that growing plants under low or high levels of Ca 2+
could increase or decrease the incidence of these disorders, respectively,
which were then named Ca 2+ deficiency disorders (Raleigh and Chucka, 1944;
Bussler, 1962; Hewitt, 1963; Shear, 1975; Chiu and Bould, 1977). Further stud-
ies have been focused on practical methods to reduce or predict the incidence
of Ca 2+ deficiency disorders in crop plants (Ferguson and Watkins, 1989;
Taylor and Locascio, 2004). More recent studies have improved our under-
standing of the mechanisms regulating Ca 2+ deficiency disorders in plants,
setting the stage for the development of more efficient control strategies (Ho
and White, 2005; Saure, 2005; Freitas and Mitcham, 2012).

15.2  Role of Ca2+ as an Essential

Plant Macronutrient
Significant improvements in our understanding of the mechanisms regulat-
ing Ca 2+ deficiency disorders in plants must follow a better understanding
of the roles of Ca 2+ at the cellular level. Calcium has a large ion radius that
facilitates ion dehydration and, consequently, binding to several anionic sub-
stances (Hauser et al., 1976; Jaiswal, 2001; Batistic and Kudla, 2010). This
property allows Ca 2+ to form noncovalent bonds within the pectin matrix of
the cell wall, contributing to cell wall structure and strength (Marschner,
1995). One type of noncovalent bond, known as a coordination bond, is
formed between Ca 2+ and oxygen or nitrogen present in pectic polysaccha-
rides (Marschner, 1995). Another type of noncovalent bond, known as an
electrostatic bond, is formed when Ca 2+ is attracted to a negatively charged
group, such as carboxylate (−COO –) on pectates and polygalacturonic acid
(Marschner, 1995). The high abundance of Ca 2+ in the cell wall has a strong

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effect not only on cell wall strength, but also on cell wall pH due to its cation
effect on the cell wall medium (Demarty et al., 1984). In addition, Ca 2+ has
also been reported to affect the synthesis of cell wall polysaccharides, such
as 1,3-β-glucan (Kauss, 1987; Brett and Waldron, 1996).
The high binding capacity of Ca 2+ makes this ion an important signal-
ing molecule in the cytosol (Hepler and Wayne, 1985; White and Broadley,
2003; Batistic and Kudla, 2010). Indeed, Ca 2+ plays an important role in
cytosolic signal transduction pathways involved in cell responses to a wide
range of biotic and abiotic factors (Scrase-Field and Knight, 2003; White
and Broadley, 2003). In the resting or quiescent state, cytosolic Ca 2+ ranges
between 100 and 200 nM, reaching about 2 µM under stimulus (Rudd and
Franklin-Tong, 1999). Changes in cytosolic Ca 2+ concentrations may take the
form of single calcium transients (Knight et al., 1996), oscillations (McAinsh
et al., 1995), or repeated spikes (Ehrhardt et al., 1996). The spatial and tem-
poral characteristics of these stimuli-specific Ca 2+ transients have become
known as Ca 2+ signatures (­Scrase-Field and Knight, 2003). Specific Ca 2+ sig-
natures have been suggested to encode information about the type and sever-
ity of the input stimulus (Dolmetsch et al., 1997), which is then decoded by
downstream components of the signaling pathway, eventually leading to the
specific and appropriate cellular response (Scrase-Field and Knight, 2003).
After raising cytosolic Ca 2+ levels, the resting state must be reestablished to
avoid Ca 2+ toxicity and, potentially, cell death. At high concentrations in the
cytosol, Ca 2+ can become toxic due to its precipitation with inorganic phos-
phate and other ionic substances, as well as its competition for binding sites
with other cations, such as Mg2+, that are required for enzyme activation and
proper cellular metabolism (Hepler and Wayne, 1985; Batistic and Kudla,
2010). For these reasons, cytosolic Ca 2+ must be under strict biochemical
and physiological control. After cytosolic oscillations, the resting state of
cytosolic Ca 2+ is reestablished by the activity of Ca-ATPases and H+/Ca 2+
exchangers, which are proteins that transport Ca 2+ from the cytosol into the
apoplast or into storage organelles in the cell (White and Broadley, 2003).
Calcium is needed at high concentrations inside cellular organelles
so it is available to be loaded into the cytosol during signaling responses,
and as a counterion to inorganic and organic anions (Jones and Bush, 1991;
White and Broadley, 2003). The vacuole is the largest pool of Ca 2+ in the cell,
with 90%–95% of the cell’s volume and 1–10 mM Ca 2+ (Bush, 1995; White
and Broadley, 2003). In the endoplasmic reticulum, Ca 2+ concentration can
range from 1 to 5 mM (Kendall et al., 1992). Chloroplasts and mitochondria
also contribute, with the Ca 2+ being stored inside these cellular organelles
at concentrations ranging between 0.1–10 µM and 0.2–1.2 µM, respectively
(Johnson et al., 1995; Logan and Knight, 2003). Inside these organelles,
Ca 2+ can be present as a free ion that can be used for cytosolic signaling
responses or as a countercation to inorganic and organic anions forming
complexes with substances such as organic acids, proteins, peptides, and

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 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

other anions (Jones and Bush, 1991; White and Broadley, 2003). Studies
have suggested that free Ca 2+ present in each organelle is ­responsible for a
specific cellular response, and that all organelles act in concert to shape a
Ca 2+ signaling response (Batistic and Kudla, 2010).
The high affinity of Ca 2+ for anionic phosphate and carboxylate groups
of lipids and proteins at the membrane surface makes Ca 2+ an important
regulator of membrane structure and function (Hanson, 1960; Hauser et al.,
1976; Clarkson and Hanson, 1980; Legge et al., 1982; Kirby and Pilbeam,
1984; Picchioni et al., 1998; Jaiswal, 2001; Hirschi, 2004; Batistic and Kudla,
2010). Calcium binding reduces membrane fluidity by tightly packing the
membrane lipids and proteins, which reduces the passive flow of monova-
lent ions such as H+, Na+, and K + (Williams, 1970; Jaiswal, 2001; White and
Broadley, 2003; Plieth, 2005). Since cytosolic Ca 2+ must be maintained at
extremely low levels to avoid toxicity, apoplastic free Ca 2+ has been sug-
gested to be the most important pool to regulate proper membrane struc-
ture and function (Hanson, 1960; Steveninck, 1965; Wallace et al., 1966;
Lund, 1970; Kirby and Pilbeam, 1984; Picchioni et al., 1998).

15.3  Symptoms of Ca2+ Deficiency Disorders in Fruit

The visual symptoms of Ca 2+ deficiency disorders are highly conserved
across different fruit species (Figure 15.1, Table 15.1). The symptoms usu-
ally start as water-soaked tissue caused by plasma membrane breakdown
that leads to cell death and tissue dehydration, resulting in dark brown and
depressed lesions on the fruit surface (Simon, 1978; Fuller, 1980; Suzuki
et al., 2000, 2003; Ho and White, 2005). Since the visual symptoms and
causes are similar in different fruit species, it is believed that the mecha-
nisms involved are also highly conserved across different species (White
and Broadley, 2003; Saure, 2005).
The symptoms of Ca 2+ deficiency can develop in different regions
depending on the species (Figure 15.1, Table 15.1). In fruit, tissue suscep-
tibility to Ca 2+ deficiency disorders is believed to be determined by Ca 2+
accumulation and cellular Ca 2+ localization (Adams and Ho, 1993; Nonami
et al., 1995; Marcelis and Ho, 1999). In that case, tissue that accumulates
less Ca 2+ is more susceptible to Ca 2+ deficiency disorders than tissue with
higher levels of Ca 2+ content (Adams and Ho, 1993; Nonami et al., 1995;
Marcelis and Ho, 1999). Some examples of Ca 2+ deficiency disorders in fruit
are described below.

15.3.1 Apple
The symptoms of Ca 2+ deficiency disorder in apple, known as bitter pit (BP),
start as water-soaked spots in the outer cortical flesh of the fruit, frequently

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Figure 15.1  Calcium deficiency disorders in fruit: (a, b) apple with BP i­nitial
water-soaked symptoms that become dark brown, depressed on the fruit
­surface; (c) blossom-end rot symptoms in tomato fruit; (d) watermelon; and
(e) bell pepper. (Panel c from Freitas and Mitcham, 2011a. Copyright ©
American Society of Plant Biologists, www.plantphysiol.org. Panel d from
Ontario Crop Integrated Pest Management (IPM), Blossom-end rot, 2009,
Queen’s Printer for Ontario, http://www.omafra.gov.on.ca/IPM/english/
cucurbits/diseases-and-disorders/blossom-end-rot.html. Copyright © 2015
Queen’s Printer for Ontario, image courtesy of John G. Strang, Department
of Horticulture, University of Kentucky. Panel e from Hochmuth, G.J., and
Hochmuth, R.C., Blossom-end rot in bell pepper: causes and prevention,
Institute of Food and Agricultural Sciences, University of Florida, Gainesville,
2012, http://edis.ifas.ufl.edu/ss497#FIGURE%203. Copyright ©  2015
University of Florida, Institute of Food and Agricultural Sciences.)

just under the skin (Figure 15.1a, Table 15.1). Later, cell dehydration and
death result in the collapse of the outermost cells, causing small dark
brown depressed lesions (pits) on the surface (Figure 15.1b). BP lesions
have also been associated with vascular elements and in severe cases may
coalesce to form larger necrotic areas (MacArthur, 1940). Although pit-
ting of the flesh also occurs, symptoms are not always visible from the out-
side. The frequency of pitting is often greater toward the calyx end of the

482
 Table 15.1  Calcium Deficiency Disorders in Fruits: Symptoms and Mechanisms Involved
Fruit Disorder Symptoms Mechanisms
Apple (A) BP Water-soaked spots 1. Reduction of total fruit Ca 2+
(A) or tissue (T, W, P) concentration (A, T, W, P)
that eventually 2. Reduction of Ca2+
becomes dark brown concentration at specific
Tomato (T) BER mainly (A) or always cellular compartments (A, T)
(T, W, P) at the distal 3. Interaction between Ca2+
fruit tissue and other nutrients (N, K+,
Mg2+) in the fruit (A)
4. Growth regulator
homeostasis at the whole-
plant and fruit-specific levels
Watermelon (W)
(A, T)
Pepper (P) 5. Environmental stress
conditions (A, T, W, P)
6. Reduction of Ca2+
concentration that results in
ROS accumulation and lipid
peroxidation in the fruit (T, P)
483
References
C alci um
Bangerth, 1979; Ferguson and
Watkins, 1989; Saure, 1996;
White and Broadley, 2003;
Freitas et al., 2010, 2013
Saure, 2001; White and
Broadley, 2003; Taylor and
Locascio, 2004; Ho and White,
2005; Saure, 2005; Freitas
D e f i c i e n cy
et al., 2011a, 2011b, 2012b,
2012c, 2014
Maynard and Hopkins, 1999
Marcelis and Ho, 1999; Aktas
et al., 2005; Hochmuth and
Hochmuth, 2012
D iso rd ers
in
Pl a nts
P OST H AR V EST RI P ENING P H Y SIOLOG Y

fruitw(Ferguson and Watkins, 1989). Fruit pitting usually takes place after
harvest, but in severe cases, it can also develop before harvest (Ferguson
and Watkins, 1989).

15.3.2 Tomato
In tomato fruit, Ca 2+ deficiency disorder was named blossom-end rot
(BER) due to its appearance: decay at the blossom end of the fruit (Figure
15.1c, Table 15.1). During BER development, blossom end tissue shows
water-soaked symptoms that become dark brown on the fruit surface at
later stages. In severe cases, BER can expand from the blossom end tis-
sue toward the calyx end tissue, affecting the whole fruit (Ho and White,
2005; Freitas et al., 2011a). During BER development, death of fruit tissue
can favor pathogen infection, which also contributes to the rot symptoms.
The initial water-soaked symptoms are believed to be caused by high mem-
brane leakage, leading to cell dehydration and death that trigger phenol
oxidation and the dark brown color development (Suzuki et al., 2000, 2003;
Ho and White, 2005). In tomato, Ca 2+ deficiency symptoms usually occur
early during fruit growth and development, when limited Ca 2+ moves into
rapidly expanding fruit (Ho and White, 2005).

15.3.3 Watermelon
Calcium deficiency disorder in watermelon is also known as BER due to its
visual symptoms, as previously described for tomato. The symptoms are
characterized by softening and shriveling of the fruit blossom end tissue,
which eventually becomes dark brown, sunken, and leathery (Figure 15.1d,
Table 15.1) (Maynard and Hopkins, 1999). Genotypes with elongated fruit
have been reported to be more susceptible to BER than round fruit geno-
types (Hammouda, 1987).

15.3.4 Pepper
The symptoms of Ca 2+ deficiency disorder in pepper are quite similar to
those of tomato fruit, which is also known as BER (Figure 15.1e, Table 15.1).
The symptoms usually occur early during fruit growth and development,
starting as water-soaked tissue in the blossom end region, which becomes
brown, with necrotic areas at later stages (Marcelis and Ho, 1999). In severe
cases, BER can expand to the entire pepper fruit (Aktas et al., 2005). Fruit
tissue close to the BER symptoms tends to lose green color faster than the
rest of the pepper. In addition, BER favors tissue infection with saprophytic

484
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

fungi and soft-rot bacteria species that enhance the rot-like appearance
(Hochmuth and Hochmuth, 2012).

15.4  Symptoms of Ca2+ Deficiency Disorders

in Leafy Vegetables
Water movement into mature leaves takes place exclusively through the
xylem, whereas water uptake into young, low-transpiring leaves takes place
though both the phloem and xylem (Taylor and Locascio, 2004). Since
Ca 2+ is only mobile in the plant through the xylem, and the rate of xylem
sap flow is controlled mainly by transpiration and growth rates, Ca 2+ accu-
mulation in old and mature leaves is much higher than in low-transpiring,
young, and enclosed leaves (Saure, 1998). In the leaf, Ca 2+ accumulation
is higher at the base and lower in the tip tissue (Barta and Tibbitts, 2000).
Therefore, the symptoms of Ca 2+ deficiency in leafy vegetables usually take
place at the tip of low-transpiring young and enclosed leaves, due to low
Ca 2+ accumulation triggered by low transpiration rates. In addition, high
tipburn incidence is always associated with environmental conditions favor-
ing plant growth (Collier and Tibbitts, 1982), which is believed to reduce
young leaf Ca 2+ concentration and increase leafy vegetables’ susceptibility
to Ca 2+ deficiency disorders. The symptoms of Ca 2+ deficiency disorders
are highly conserved among leafy vegetables, which are characterized by
stunted growth, curling, and the appearance of brown spots and necrosis
associated with phenol oxidation of young leaves, leaf tips, and primarily
meristematic cells (Figure 15.2, Table 15.2) (Bussler, 1962; Simon, 1978;
Saure, 1998; Koike and Smith, 2010). Some examples of Ca 2+ deficiency
disorders in leafy ­vegetables are described below.

15.4.1 Lettuce
Calcium deficiency disorder in lettuce, known as tipburn, is characterized
by dark brown lesions and necrosis on the margins of young developing
leaf tips (Figure 15.2a, Table 15.2) (Saure, 1998; Koike and Smith, 2010). In
some genotypes, tipburn is first seen on the small veins along the margin
of young leaves. Inner and younger leaves are more susceptible to tipburn
incidence than outer and older leaves (Koike and Smith, 2010).

15.4.2 Cauliflower
Calcium deficiency disorder in cauliflower, also called tipburn, is charac-
terized by a light brown coloration and tip necrosis of young inner wrapper

485
P OST H AR V EST RI P ENING P H Y SIOLOG Y

leaves that enclose the cauliflower head (Figure 15.2b, Table 15.2) (Rosen,
1990; Koike and Smith, 2010). In severe cases, Ca 2+ deficiency symptoms
can also result in curd discoloration due to secondary pathogen infection
(Maynard et al., 1981).

15.4.3 Artichoke
Calcium deficiency disorder in artichoke is also known as tipburn and is
characterized by brown discoloration and the death of leaf margins. In
addition, immature flower buds can also develop black lesions along the

Figure 15.2  Calcium deficiency development in leafy vegetables. Tipburn

development in (a) lettuce, (b) cauliflower, and (c) artichoke, and (d) black-
heart in celery. (From Koike, S., and Smith, R., Calcium deficiency disorders
hit vegetable crops in central coast, Regents of the University of California,
Agriculture and Natural Resources, Oakland, CA, 2010, http://ucanr.org/
blogs/blogcore/postdetail.cfm?postnum=3407. Copyright © 2015 Regents
of the University of California, Agriculture and Natural Resources.)

486
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Table 15.2  Calcium Deficiency Disorders in Vegetables: Symptoms and

Mechanisms Involved
Vegetable Disorder Symptoms Mechanisms References
Lettuce Tipburn Tip necrosis of Low Ca2+ Saure, 1998;
young leaves accumulation White and
and meristems in young leaf Broadley,
tips and 2003;
meristems Koike and
Smith, 2010
Cauliflower Rosen, 1990;
Koike and
Smith, 2010
Artichoke Francois
et al., 1991;
Koike and
Smith, 2010
Celery Blackheart Light to dark White and
brown speckling, Broadley,
lesions and 2003;
necrosis on the Koike and
margins of Smith, 2010
developing leaf
tips deep within
the central
growing region

upper tips and edges of the flower bracts. These symptoms appear mainly
on bracts in the inner whorls of the bud (Figure 15.2c, Table 15.2) (Francois
et al., 1991; Koike and Smith, 2010). In severe cases, Ca 2+ -deficient bracts
can also be infected by pathogens (Francois et al., 1991).

15.4.4 Celery
The Ca 2+ deficiency disorder in celery is known as blackheart and is char-
acterized by light to dark brown speckling, lesions, and necrosis on the
margins of developing leaf tips deep within the central growing region
(Figure  15.2d, Table 15.2) (Koike and Smith, 2010). During plant growth
and development, the blackheart symptoms may turn black and expand
outward from the inner plant tissues (Koike and Smith, 2010).

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15.5  Potential Mechanisms Regulating

Ca2+ Deficiency Disorders
After years of study, it is believed that plant susceptibility to Ca 2+ deficiency
disorders is affected by genetic and environmental factors regulating tissue
Ca 2+ content and cellular Ca 2+ distribution (Figure 15.3) (Bangerth, 1979;
Ferguson and Watkins, 1989; Francois et al., 1991; Saure, 1998, 2005; Taylor
and Locascio, 2004; Ho and White, 2005; Freitas and Mitcham, 2012).

15.5.1  Total Tissue Ca2+ Content

Calcium accumulation in leaf and fruit tissues is regulated at the whole plant
and at the leaf- and fruit-specific levels (Figure 15.3). At the whole-plant
level, Ca 2+ movement into the leaf or fruit is determined first by Ca 2+ avail-
ability to the plant and root Ca 2+ uptake activity (Taylor and Locascio, 2004).

Environment

Genotype

Plant Ca2+ uptake

Fruit Ca2+ uptake

Ca2+/other nutrients

Growth regulators

Ca2+ movement in
the fruit

Cellular Ca2+
distribution
Ca2+ deficiency
disorders
Reactive oxygen
species (ROS)

Figure 15.3  Potential mechanisms regulating Ca2+ deficiency disorders in

fruit and vegetables.

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 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Most of the root Ca 2+ uptake is believed to take place passively through the
apoplast at the root tip and lateral root growth regions where the Casparian
band is not present (White, 2001; Taylor and Locascio, 2004). In that case,
root growth enhances Ca 2+ uptake by increasing the number of root tips and
lateral roots that lead to higher apoplastic Ca 2+ movement into the root sys-
tem (White, 2001). Although root Ca 2+ uptake may also take place through
the symplastic pathway, the complex mechanisms that have evolved to
strictly regulate and maintain low levels of cytosolic Ca 2+ suggest other-
wise (White, 2001). In addition, root Ca 2+ uptake is also affected by uptake
­competition between Ca 2+ and other ions available in the soil, as will be dis-
cussed later in this chapter on section 15.5.3. (Bangerth, 1979; Ferguson
and Watkins, 1989; Saure, 2005). Therefore, soil nutrient composition also
influences root Ca 2+ uptake and, consequently, leaf and fruit susceptibility
to Ca 2+ deficiency disorders.
After Ca 2+ is taken up by the roots, it moves in the xylem vessels
toward the leaves and fruit by mass flow in response to the water potential
gradient generated by leaf and fruit transpiration and growth (Saure, 1998;
Taylor and Locascio, 2004). Mature leaves have higher transpiration rates
than young leaves and fruit, which explain their higher Ca 2+ accumulation
(Saure, 1998; Ho and White, 2005; Freitas et al., 2011b). Accordingly, low rel-
ative humidity increases Ca 2+ uptake into mature high-transpiring leaves,
but reduces in low-transpiring fruit and the inner leaves of leafy vegetables
with a closed growing point (Olle and Bender, 2009). In addition, mature
leaves receive water exclusively from the xylem, whereas young leaves and
fruit receive water from both xylem and phloem vessels (Ho and White,
2005; Freitas et al., 2011b). Considering that Ca 2+ moves in the plant only
through the xylem, water uptake from the phloem further decreases the
capacity to uptake Ca 2+ through the xylem sap into young leaves and fruit,
compared to mature leaves. Similar behavior is also observed in other sink
organs, such as meristems that have low transpiration rates and receive
water from both the phloem and xylem, making sink organs highly sus-
ceptible to Ca 2+ deficiency disorders (Saure, 1998; Koike and Smith, 2010).
Studies have shown that increasing transpiration of sink organs, without
changing whole-plant transpiration, is more efficient to increase Ca 2+ con-
tent in these organs than increasing Ca 2+ availability in the soil (Tadesse
et  al., 2001). In addition, reducing whole-plant transpiration either by
decreasing water vapor pressure deficit (WVPD) or by spraying the plant
with abscisic acid (ABA) can reduce xylem sap movement toward mature
leaves and increase xylemic flow toward low-transpiring fruit, which has
been shown to increase fruit Ca 2+ accumulation and decrease fruit suscep-
tibility to Ca 2+ deficiency disorders (Guichard et  al., 2005; Freitas et al.,
2011b, 2014; Barickman et al., 2014).
Plant water stress has been widely reported to increase Ca 2+ defi-
ciency disorder development in both low-transpiring leaves and fruit

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(Saure, 1998; Ho and White, 2005; Koike and Smith, 2010). Water stress,
triggered by low water availability or high salinity conditions, reduces
the water potential gradient between roots and low-transpiring leaves and
fruit, consequently decreasing Ca 2+ uptake and increasing tissue suscep-
tibility to Ca 2+ deficiency disorders (Taylor and Locascio, 2004; Freitas
and Mitcham, 2012). This effect is further enhanced by Ca 2+ immobility
through the phloem, ensuring that the high levels of Ca 2+ accumulated in
older leaves are not redistributed to low-transpiring Ca 2+ -deficient tissues
(White and Broadley, 2003).
Although transpiration and growth are important factors determin-
ing xylem sap flow and Ca 2+ uptake, the number of functional xylem vessels
and the hydrostatic gradient required for xylem sap movement can also
affect Ca 2+ content in sink organs (Ho et al., 1993; Tadesse et al., 2001;
Dražeta et  al., 2004a; Bondada et al., 2005; Ho and White, 2005; Freitas
et  al., 2011b). Accordingly, studies have shown that the number of func-
tional xylem vessels and Ca 2+ accumulation in the fruit decrease simulta-
neously during fruit growth and development (Nonami et al., 1995). High
numbers of functional xylem vessels have also been associated with high
levels of fruit Ca 2+ uptake and low fruit susceptibility to Ca 2+ deficiency
disorders (Freitas et al., 2011b). Besides a high number of functional xylem
vessels, the hydrostatic gradient in the xylem vessels may also be required
for xylem sap movement into distal fruit tissues (Bondada et al., 2005),
contributing to Ca 2+ accumulation and reducing susceptibility to Ca 2+ defi-
ciency disorders.

15.5.2  Cellular Regulation of Ca2+ Partitioning

and Distribution
Mechanisms regulating cellular Ca 2+ partitioning have recently been
reported in the literature (Park et al., 2005; Conn et al., 2011; Freitas et al.,
2011a; Wu et al., 2012). As previously described, Ca 2+ is required at spe-
cific concentrations in each cellular compartment (White and Broadley,
2003). Therefore, abnormal changes in cellular Ca 2+ partitioning can
potentially result in a cell-localized Ca 2+ deficiency, cell death, and Ca 2+
deficiency symptom development (Figures 15.3 and 15.4). Indeed, stud-
ies have shown that abnormal regulation of cellular Ca 2+ partitioning
represents an important mechanism determining tissue susceptibility to
Ca 2+ deficiency disorders in plants (Park et al., 2005; Freitas et al., 2011a)
(Figure 15.4).
Studies have shown that mutant plants lacking the expression Ca 2+/
proton antiporters CAX1 and CAX3 in the tonoplast have higher apoplas-
tic free Ca 2+ content than wild-type plants, suggesting that CAX genes

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are indeed important mechanisms regulating cellular Ca 2+ partitioning

(Conn et al., 2011). Accordingly, attempts to increase fruit Ca 2+ content by
­enhancing the expression of a tonoplast CAX resulted not only in higher
fruit Ca 2+ accumulation, but also in higher fruit susceptibility to Ca 2+ defi-
ciency disorder (Park et al., 2005) (Figure 15.4). In these studies, the
Arabidopsis CAX1 gene without the N terminus regulatory region (sCAX1)
was inserted into the tomato genome under the control of the cell cycle
promoter cdc2a (Park et al., 2005). The cdc2a::sCAX1 construct resulted
in high sCAX1 expression during cell division, which produces a consti-
tutively active protein (Park et al., 2005). The sCAX1-transformed fruit
had about twofold higher total tissue Ca 2+ content and higher vacuolar
Ca 2+ accumulation than wild-type fruit (Figure 15.4). However, sCAX1-
expressing fruit also showed lower cytosolic and apoplastic Ca 2+ concen-
trations that possibly resulted in the observed higher plasma membrane
leakage, cell plasmolysis, and 100% incidence of Ca 2+ deficiency symp-
toms in fruit, compared to wild-type fruit that did not develop Ca 2+ defi-
ciency symptoms (Freitas et al., 2011a) (Figure 15.4). Coexpression of an
endoplasmic reticulum Ca 2+ binding protein known as calreticulin has
been shown to mitigate Ca 2+ deficiency symptoms observed in sCAX1-
expressing fruit (Wu et al., 2012). Although the authors suggested that

Healthy fruit

Normal fruit Ca uptake

2+
Limited fruit Ca2+ uptake

CAX CAX PME PME

Fruit Fruit Cell wall Cell wall

[Ca2+] [Ca2+] [Ca2+] [Ca2+]

Apopl. sol. Apopl. sol. Apopl. sol. Apopl. sol.

[Ca2+] [Ca2+] [Ca2+] [Ca2+]

M. leakage M. leakage M. leakage M. leakage

No BER BER BER BER

Figure 15.4  Potential mechanisms regulating cellular Ca2+ distribution and

fruit and vegetable susceptibility to Ca2+ deficiency disorders. CAX, Ca2+/
proton antiporters; PME, pectin methylesterase; Apopl. sol., apoplastic soluble;
M. leakage, membrane leakage.

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coexpression of the endoplasmic reticulum calreticulin reduced fruit

susceptibility to Ca 2+ deficiency disorders by altering the cellular Ca 2+
content and distribution within the plant matrix (Wu et al., 2012), more
studies are required to further understand the mechanisms involved. It
is possible that calreticulin could mitigate Ca 2+ deficiency symptoms in
sCAX1-expressing fruit by mechanisms unrelated to cellular Ca 2+ distri-
bution (Qiu et al., 2012).
Plants have evolved with cell walls as defensive barriers, conduits for
information, and sources of signaling molecules and developmental cues
(Bacic et al., 1988; O’Neill et al., 1990; Carpita and Gibeaut, 1993; Ridley
et al., 2001). Cell wall composition and, consequently, its Ca 2+ binding capac-
ity can vary widely from plant to plant (Sorensen et al., 2010). Accordingly,
dicotyledeonous plants have higher cell wall Ca 2+ binding capacity and
Ca 2+ requirements than monocotyledonous plants (Islam et al., 1987;
Kirkby and Pilbeam, 1984; White and Broadley, 2003). Considering that
the cell wall contains, on average, 70% of the total plant tissue Ca 2+ content
(Demarty et al., 1984; Freitas et al., 2010), small changes in cell wall Ca 2+
binding capacity can potentially have a great impact on Ca 2+ availability
in other pools of the cell. Indeed, recent studies suggest that increasing
cell wall Ca 2+ binding capacity can reduce the levels of Ca 2+ available in
other pools in the cells, resulting in a cell-­localized Ca 2+ deficiency, cell
death, and Ca 2+ deficiency symptom development (Freitas et al., 2012b)
(Figure  15.4). Silencing tomato fruit pectin methylesterase (PME) genes
reduced the number of carboxyl groups in the cell wall matrix, and there-
fore reduced cell wall Ca 2+ content (Freitas et  al., 2012c) (Figure 15.4).
These PME-silenced fruit had increased apoplastic water-soluble Ca 2+,
lower membrane leakage, and reduced susceptibility to Ca 2+ deficiency
disorders (Figure 15.4). These studies suggest that regulation of cellular
Ca 2+ distribution, either by the activity of enzymes involved in cellular
Ca 2+ transport or by Ca 2+ binding to the cell wall, is a potential mechanism
regulating tissue susceptibility to Ca 2+ deficiency disorders (Figure 15.4).

15.5.3  Other Nutrients

15.5.3.1 Nitrogen
Nitrogen (N) is commonly applied at high rates in the soil to stimulate
plant growth and yield (Bramlage and Weis, 2004). However, excessive
N levels might increase fruit susceptibility to Ca 2+ deficiency disorders
(Figure 15.3) (Raese and Drake, 1997; Dris et al., 1998). Besides the total
amount of N, the form of N that is used can also influence fruit quality.
Soil fertilization with ammonium rather than nitrate nitrogen substantially

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reduced Ca 2+ accumulation in apples and greatly increased the incidence

of BP (Ludders, 1979), since ammonium is antagonistic to Ca 2+ uptake by
roots (Shear and Faust, 1971). However, since ammonium can be rapidly
converted to nitrate in the surface layers of the soil, it is believed that
the total amount of N being applied to the soil is a much more important
concern to Ca 2+ deficiency disorders than is the form in which N is being
applied (Bramlage et al., 1980).
High N fertilizer doses lead to substantial vegetative growth
(Bramlage and Weis, 2004). Since Ca 2+ moves with water in the transpira-
tion stream, greater leaf area will shunt more water and Ca 2+ to the leaves
rather than fruit, reducing the Ca 2+ content in the fruit (Saure, 2005).
Nitrogen fertilization also has an indirect effect on Ca 2+ deficiency dis-
order development by increasing fruit size (van Schreven, 1961; Ho and
White, 2005).
Fruits with high levels of N also show higher respiration and eth-
ylene production rates after harvest, reflecting advanced ripening and
senescence stages (Fallahi et al., 2010), and this possibly accelerates the
mechanisms leading to Ca 2+ deficiency disorders (Lötze and Theron, 2006;
Lötze et al., 2010).

15.5.3.2  Potassium and Magnesium

Excessive potassium (K ) and magnesium (Mg2+) fertilization has been
+

found to increase the incidence of BP in apples (Ferguson and Watkins,

1989) and BER in tomatoes (Ho et al., 1993) (Figure 15.3). Concentrations
of K +, Mg2+, and Ca 2+ in roots and mature leaves of cultivated plants are
generally high, but although K + is readily redistributed within the plant,
concentrations of Mg2+, and especially Ca 2+, are often low in phloem-fed tis-
sues such as fruits and young leaves (Karley and White, 2009). In addition,
at the onset of cell expansion in fruits, xylem dysfunction often begins and
might lead to reduced fruit uptake of Ca+2 , while the function of the phloem
remains unchanged relative to the xylem flow, favoring the supply of K +
(and to a lesser extent Mg2+) (Lang, 1990; Dražeta et al., 2004a).
High K +/Ca 2+, Mg2+/Ca 2+, or (K + + Mg2+)/Ca 2+ ratios in the fruit
have been used to predict BP in apples (Vang-Petersen, 1980; Ferguson
and Watkins, 1989; Argenta and Suzuki, 1994; Nachtigall and Freire,
1998; Lanauskas and Kvikliene, 2006; Amarante et al., 2006a, 2006b,
2013). Potassium and Mg2+ compete with Ca 2+ for binding sites at the
plasma membrane surface, but these elements cannot replace the role of
Ca 2+ in membrane structure and stability (Schonherr and Bukovac, 1973;
Yermiyahu et al., 1994). Consequently, less Ca 2+ will be bound to the plasma
membrane, which will become leakier, eventually leading to plasmolysis,

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membrane breakdown, and cell death, during the manifestation of Ca 2+

deficiency disorders (Freitas et al., 2010, 2013).

15.5.3.3 Boron
Boron (B) is a micronutrient element that plays a key role in reproductive
processes such as pollen germination and pollen tube growth (Dickinson,
1978), and application of B increases fruit set and yield in B-deficient
apple and pear trees (Wojcik and Wojcik, 2003; Wojcik and Treder, 2006;
Wojcik et al., 2008). Boron has a synergistic effect with Ca 2+ and helps to
increase the Ca 2+ concentration in fruit (Dickson et al., 1973; Bramlage
et al., 1980; Wojcik et al., 1999; Wojcik and Wojcik, 2003; Bramlage and
Weis, 2004) and reduce BP incidence in apples (Faust and Shear, 1968;
Granelli and Ughini, 1990; Wojcik et al., 1999; Sen et al., 2010) and inter-
nal browning in pears (Wojcik and Wojcik, 2003; Mielke and Chaplin,
2008) (Figure 15.3). Boron might suppress the activity of indole-3-acetic
acid (IA A) oxidase (Marschner, 1995), leading to an increase in IA A
that promotes acropetal movement of Ca 2+ (Dela Fuente et al., 1986) and
stimulates xylem tissue differentiation in the pedicel of the fruit (Dražeta
et al., 2004b), thereby improving Ca 2+ uptake by fruits. Soil fertilization
with B also increased the dry weight of fine roots in apple trees (Wojcik
et  al., 2008), where most of the Ca 2+ uptake is believed to take place
(White, 2000, 2001; Taylor and Locascio, 2004). This might also con-
tribute to increased Ca 2+ content of leaves and fruits in plants fertilized
with B. Boron also plays an important role in maintaining both the struc-
tural and functional properties of membranes (Cakmak and Römheld,
1977), thereby reducing internal browning disorders in pear fruit (Xuan
et al., 2001, 2005; Mielke and Chaplin, 2008).

15.5.3.4 Phosphorus
Mulder (1952) reported that BP in apple fruit was associated with low P con-
tent. However, several authors reported that BP was associated with high
P content (Brown, 1926; Oberly and Kenworthy, 1961; Sharples, 1964). The
high P content of the affected fruit is not surprising if one takes into account
the observation that mineral elements move into the pitted tissue (Chamel
and Bossy, 1981).
Apple trees supplied annually at bloom with P (with 20 g P per tree as
ammonium polyphosphate, and receiving adequate fertigation applications
of N, K+, and B) had fruit with reduced incidence of water core (also a Ca 2+
deficiency disorder) at harvest, higher resistance to browning of cut slices,
reduced membrane leakage, and elevated antioxidant content after cold-air
storage (Neilsen et al., 2008). This indicates a role for P in the maintenance
of apple fruit membrane stability and cellular energetics that might aid in

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 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

reducing the incidence of Ca 2+ deficiency disorders in apples (Bramlage

et al., 1980) (Figure 15.3).

15.5.4  Reactive Oxygen Species

It has been suggested that Ca 2+ deficiency is not the cause of BER, but a
result of it, with BER caused by stress conditions leading to high levels of
reactive oxygen species (ROS) that disintegrate cellular membranes, result-
ing in BER symptoms in fruit tissue (Saure, 2014). Indeed, previous studies
have shown higher levels of ROS, such as superoxide radicals, hydroxyl
radicals, and singlet oxygen (O2), in fruit tissue with BER (Aktas et al.,
2003, 2005; Turhan et al., 2006; Mestre et al., 2012). However, an extensive
study focusing on the relationship between oxidative metabolism and BER
development revealed that reducing fruit Ca 2+ concentration also reduced
the activity of the main enzymes responsible for ROS detoxification, lead-
ing to H 2O2 accumulation, lipid peroxidation, and BER symptom develop-
ment (Mestre et al., 2012). Therefore, Ca 2+ can inhibit BER development
directly by binding to phospholipids and proteins on cellular membranes,
and by stimulating the activity of enzymes required for ROS detoxification.
Considering that environmental stress conditions can increase ROS levels
independent of fruit Ca 2+ content (Saure, 2001), it is possible that neither
Ca 2+ nor ROS alone can fully explain Ca 2+ deficiency disorder development,
but the interaction between Ca 2+ and ROS concentrations in the tissue. In
addition, studies show that regulation of cellular Ca 2+ distribution plays an
important role in fruit susceptibility to BER (Freitas et al., 2011a, 2012b),
suggesting that the combined Ca 2+/ROS effect on Ca 2+ deficiency disor-
ders may also be cellularly compartmentalized in the apoplast. In that case,
Ca 2+ deficiency disorders may develop only if Ca 2+ levels in the apoplast
are not able to counteract the ROS effects on membrane lipid peroxida-
tion (Figure 15.3). Although these studies aided our understanding of the
mechanisms involved in Ca 2+ deficiency disorders, future studies should
better characterize the combined effects of different Ca 2+/ROS ratios in
different cellular compartments on Ca 2+ deficiency disorders.

15.5.5  Growth Regulators

Although many growth regulators have been associated with Ca 2+ defi-
ciency disorders through specific mechanisms (Saure, 1996, 1998, 2001,
2005; Ho and White, 2005), the final tissue susceptibility to these disorders
is likely determined by the combined effects of various growth regulators
on total tissue Ca 2+ accumulation and cellular Ca 2+ distribution (Freitas and
Mitcham, 2012) (Figure 15.3).

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15.5.5.1  Growth Regulators Affecting Total Tissue Ca2+ Content

15.5.5.1.1  Growth Regulators Influencing Root Growth and Ca2+ Uptake
Activity  Root Ca 2+ uptake can be determined by a growth regulator’s
effect on root growth and activity. Studies have suggested that basipetal
auxin transport and root auxin content are responsible for increases in
root growth and activity (Dewitte and Murray, 2003; Yang et al., 2004;
Chapman and Estelle, 2009). Accordingly, maintaining auxin transport
and increasing root auxin content have been shown to increase root activ-
ity and Ca 2+ uptake, resulting in higher leaf and fruit Ca 2+ accumulation
(Steenkamp and de Villiers, 1979; Yang et al., 2004). Although cytokinins
have been reported to promote cell differentiation in the roots (Chapman
and Estelle, 2009), treating plants with cytokinins can also increase
root Ca 2+ uptake by enhancing root affinity for Ca 2+ (Yang et al., 2008).
Gibberellins (GAs) and ABA have been shown to inhibit cell division and
growth by enhancing the transcription of cell cycle inhibitors (Wang
et al., 1998; Achard et al., 2009). Accordingly, GAs have been suggested
to reduce root Ca 2+ uptake, decreasing Ca 2+ accumulation in fruit, meri-
stems, and leaves (Cohen and Greene, 1989; Monge et al., 1994; Saure,
2005). These studies show that root growth and activity are tightly reg-
ulated by several growth regulators (Ubeda-Tomás et al., 2012), affect-
ing root Ca 2+ uptake, plant Ca 2+ content, and consequently, leaf and fruit
­susceptibility to Ca 2+ deficiency disorders.

15.5.5.1.2  Growth Regulators and Xylem Vessel Development  Since Ca 2+

moves in the plant exclusively through the xylem vessels, higher numbers
of functional xylem vessels can favor Ca 2+ movement into leaves and fruit,
potentially reducing the susceptibility of these organs to Ca 2+ deficiency
disorders (White and Broadley, 2003; Saure, 2005). Growth regulators are
known to control vascular tissue differentiation (Aloni, 1987). Xylem ves-
sel differentiation is triggered by basipetal auxin transport in the plant,
leaf, and fruit (Bustan et al., 1995; Saure, 2005), which can be enhanced
by the presence of cytokinins (Aloni, 2001). Accordingly, plants treated
with the auxin transport inhibitor 2,3,5-triiodobenzoic acid have reduced
Ca 2+ uptake into sink tissues (Banuelos et al., 1987; Cutting and Bower,
1989). Other studies suggest that xylem differentiation is triggered by high
auxin/GA ratios, whereas low ratios have been associated with the devel-
opment of phloem (Aloni, 1987, 2001; Saure, 2005). Gibberellins are also
known to trigger cell expansion, which has been suggested to result in the
constriction of xylem vessels during stages of rapid fruit expansion, leading
to reduced fruit Ca 2+ uptake and increased fruit susceptibility to Ca 2+ defi-
ciency disorders (Dražeta et al., 2004a; Saure, 2005; Freitas et al., 2012a).
Accordingly, treating tomato plants with GA has been shown to decrease,
whereas treating plants with prohexadione-calcium, a GA ­ biosynthesis

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inhibitor, has been shown to maintain the number of functional xylem ves-
sels during fruit growth and development, which was highly correlated with
fruit Ca 2+ uptake and susceptibility to Ca 2+ deficiency disorders (Freitas
et  al., 2012a). Treating tomato plants with ABA has also been shown to
maintain higher numbers of functional xylem vessels and higher xylem/
phloem ratios of water uptake into the fruit, which resulted in higher Ca 2+
uptake and lower fruit susceptibility to Ca 2+ deficiency disorders (Freitas
et al., 2011b, 2014; Barickman et al., 2014).

15.5.5.1.3  Growth Regulators Controlling Transpiration and Growth  Leaf

transpiration is known to be regulated by stomatal opening in response to
ABA content in the leaf (Li et al., 2000). Studies have shown that treat-
ing whole plants with ABA can specifically trigger leaf stomatal closure,
decreasing whole-plant leaf transpiration without affecting fruit transpira-
tion (Freitas et al., 2011b, 2014). Leaf stomatal closure decreased xylem
sap and Ca 2+ uptake into mature leaves and increased xylem sap and Ca 2+
uptake into the fruit, reducing fruit susceptibility to Ca 2+ deficiency disor-
ders under Ca 2+ stress conditions (Freitas et al., 2011b, 2014). Accordingly,
conditions that favor high mature leaf transpiration, plant water loss, and
plant water stress have been shown to reduce fruit xylem sap and Ca 2+
uptake and increase fruit susceptibility to Ca 2+ deficiency disorders (Abdal
and Suleiman, 2005; Guichard et al., 2005; Freitas et al., 2011b, 2014;
Barickman et al., 2014).
Fruit and leaf growth rates are determined by the combined pro-
cesses of cell division and expansion, which are controlled by growth
regulator homeostasis in the tissue (Gillaspy et al., 1993). Studies have
suggested that plant organs are usually more susceptible to Ca 2+ defi-
ciency disorders when higher growth rates are combined with lower tis-
sue Ca 2+ uptake, which leads to dilution of tissue Ca 2+ content (Taylor
and Locascio, 2004; Ho and White, 2005; Saure, 2005). Accordingly, distal
leaf and fruit tissues are believed to have higher susceptibility to Ca 2+
deficiency disorders because of their higher growth rates and lowest total
Ca 2+ content (Saure, 1998, 2005; Barta and Tibbitts, 2000). Gibberellins
trigger cell expansion (Gillaspy et al., 1993; Achard et al., 2009), which
has been suggested to decrease Ca 2+ uptake and dilute Ca 2+ content in
leaves and fruit, increasing tissue susceptibility to Ca 2+ deficiency disor-
ders (Saure, 1998, 2005). The inhibition of Ca 2+ deficiency disorders by
ABA has also been attributed to its antagonistic effect on GA responses
that limit leaf and fruit Ca 2+ uptake (Saure, 1998; Freitas et al., 2011b;
Freitas and Mitcham, 2012).
Amarante et al. (2003) have shown that spraying apple trees at full
bloom with increasing concentrations (up to 20 mg L −1) of thidiazuron
(TDZ; N-phenyl-N′-1,2,3-thiadiazol-5-ylureia), which has c­ytokinin-like
activity, to promote fruit set, increases vegetative growth and consequently

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reduces Ca 2+ concentrations in the fruit. According to the authors, the

intense promoting growth effect of TDZ on vegetative tissues (terminal
buds and growing leaves) might increase the accumulation of nutrients
into these organs, especially Ca 2+, to the detriment of growing fruit. On the
other hand, Greene (1995) reported a higher BP incidence at harvest when
apple trees were sprayed 18 days after full bloom with 15 mg L −1 TDZ, to
promote fruit thinning. Therefore, the treatment with TDZ to reduce crop
load might promote fruit growth and dilute fruit Ca 2+ content, increasing
the risk of BP (Greene et al., 1990; Elfving and Cline, 1993; Greene, 1993).

15.5.5.2  Growth Regulators Influencing Cellular

Ca2+ Distribution
At the cellular level, growth regulators have been demonstrated to affect
tissue susceptibility to Ca 2+ deficiency disorders by regulating cellular
Ca 2+ partitioning and possibly by modulating cell responses to symptom
development (Ho and White, 2005). Both auxins and GAs are known to
trigger cell enlargement and tissue expansion (Saure, 1996, 2001, 2005;
Perrot-Rechenmann, 2010). The combined effect of high cell enlarge-
ment rates and restricted tissue Ca 2+ uptake triggered by auxin and GAs
could result in excessive cell expansion, leading to increased membrane
leakage and Ca 2+ deficiency symptom development (Ho and White, 2005).
Recently, studies have shown that spraying tomato plants with GAs or a
GA biosynthesis inhibitor (prohexadione-calcium) resulted in 100% or 0%
incidence of Ca 2+ deficiency symptoms, respectively (Freitas et al., 2012a).
More detailed analyses revealed that GA treatment increased the expres-
sion of genes coding for Ca 2+ transport proteins that lead to Ca 2+ movement
into cellular storage organelles, reduced apoplastic water-soluble Ca 2+, and
increased fruit tissue membrane leakage (Freitas et al., 2012a). These data
suggest that GAs trigger abnormal cellular Ca 2+ distribution that increases
tissue susceptibility to Ca 2+ deficiency disorders. ABA is believed to act as
an antagonist of many GA responses (Saure, 2001). Indeed, studies have
shown that ABA not only increases total fruit tissue Ca 2+ uptake, but also
may act as a GA antagonist, maintaining higher levels of apoplastic water-
soluble Ca 2+ and decreasing fruit susceptibility to Ca 2+ deficiency disorders
(Freitas et al., 2011b, 2014).
Ethylene is known to accelerate fruit ripening and senescence-
related processes, and possibly accelerate the mechanisms leading to Ca 2+
deficiency symptom development (Lötze and Theron, 2006; Lötze et  al.,
2010). Ethylene could accelerate Ca 2+ deficiency disorders by increas-
ing the expression and activity of PMEs, increasing Ca 2+ binding to the
cell wall and reducing Ca 2+ availability in other pools in the cell (Freitas
et  al., 2010, 2013). Later, during fruit ripening and softening, increasing

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 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

the activity of cell wall–degrading enzymes could release Ca 2+ from the

cell wall matrix back into other pools in the cell, reducing tissue suscepti-
bility to Ca 2+ deficiency disorders (Freitas et al., 2010). Accordingly, Ca 2+
deficiency symptom development is usually restricted to a short period of
time during apple development, which is highly correlated with an increase
in PME expression and activity (Freitas et al., 2010). Previous studies
have also shown that ethylene increases plasma membrane permeability
(Candan et al., 2008), which could also enhance tissue susceptibility to Ca 2+
deficiency disorders under conditions of low Ca 2+. Other growth regulators
not mentioned here may also affect plant tissue susceptibility to Ca 2+ defi-
ciency disorders. However, the mechanisms involved remain to be explored
and understood.

15.5.5.3  Growth Regulator Effect on Oxidative Metabolism

It has recently been suggested that GAs increase the susceptibility to
Ca 2+ deficiency disorders by downregulating ROS scavenging enzymes
such as superoxide dismutase, catalase, and ascorbate peroxidase (Kwak
et al., 2006; Saure, 2014), and by stimulating the destruction of the growth-
inhibiting DELLA proteins, which normally cause ROS levels to remain
low under environmental stress (Achard et al., 2008; Saure, 2014). Since
ABA is an antagonist to GAs, it has been suggested that ABA inhibition of
GA-induced Ca 2+ deficiency disorders can be due to its effect on increasing
plant and fruit stress tolerance (Saure, 2014).
Growth regulators such as brassinosteroids have been reported to
increase plant stress tolerance by increasing the activity of ROS scaveng-
ing enzymes (Schenabel et al., 2001; Liu et al., 2009). Although there are
no studies showing the effect of brassinosteroids on Ca 2+ deficiency disor-
ders, the role of these growth regulators suggests a possible inhibition of
fruit susceptibility to Ca 2+ deficiency disorders by enhancing ROS scav-
enging (Freitas and Mitcham, 2012). Similarly, salicylic acid has also been
shown to make plants more tolerant to high salinity conditions, reducing
membrane permeability of leaf tissue (Stevens et al., 2006). Considering
that salinity conditions are known to increase ROS levels in plants, sali-
cylic acid, similar to brassinosteroids, can act on ROS scavenging and may
have an inhibitory effect on Ca 2+ deficiency disorders. Other studies have
shown that high levels of methyl jasmonate increase fruit susceptibility
to Ca 2+ deficiency disorders (Rudell et al., 2005) and fruit phenol content
(Rudell et al., 2002). Since Ca 2+ deficiency symptoms involve phenol oxi-
dation (Dekock et al., 1980; Casado-Vela et al., 2005), methyl jasmonate
may increase fruit susceptibility to Ca 2+ deficiency disorders by increas-
ing phenol content that favors phenol oxidation in fruit tissue (Freitas and
Mitcham, 2012).

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15.6  Possible Control Strategies

The information presented in this review demonstrates that fruit and leafy
vegetable susceptibility to Ca 2+ deficiency disorders is determined by genetic
and environmental factors affecting tissue Ca 2+ content and cellular Ca 2+ dis-
tribution. Therefore, prior to developing control strategies, one should first
identify the genetic and environmental factors leading to limited tissue Ca 2+
content or abnormal cellular Ca 2+ distribution in order to develop specific
strategies that can effectively control Ca 2+ deficiency disorders.

15.6.1  At the Tissue Level

Adequate soil Ca 2+ availability and root Ca 2+ uptake are usually the first
strategy to ensure Ca 2+ accumulation in leaf and fruit tissue. Selection of
genotypes and rootstocks with higher root Ca 2+ uptake activity can poten-
tially reduce susceptibility of new crop plants to Ca 2+ deficiency disorders.
Adequate use of other nutrients can avoid root uptake competition with
Ca 2+ or excessive vegetative growth that leads to higher Ca 2+ movement
into mature leaves and away from low-transpiring young leaves and fruit.
Calcium uptake and movement into low-transpiring young leaves and fruit is
negatively correlated to plant water stress. Therefore, adequate plant water-
ing can also facilitate root Ca 2+ uptake and movement into low-­t ranspiring
young leaves and fruit.
Although Ca 2+ is believed to move in the plant exclusively through the
xylem vessels, the development of technologies that allow Ca 2+ movement
through the phloem could allow Ca 2+ movement from older leaves into low-
transpiring young leaves and fruit, which receive most of their water from
the phloem. One example of such a technology was developed for boron
(B), which is also considered to be phloem immobile or to have only limited
phloem mobility in higher plants (Brown and Hu, 1996). Studies revealed
that B is mobile in the phloem of some plant species when it forms stable
complexes with sorbitol (Brown and Hu, 1996). Based on these studies,
tobacco plants were engineered to synthesize high amounts of sorbitol,
which resulted in a marked increase in B mobility in the plant and increased
plant growth and yield when grown with limited or interrupted soil B sup-
ply, compared to wild-type plants (Brown et al., 1999). Analyses revealed
that transgenic plants were able to remobilize B present in mature leaves
through the phloem into sink organs, which was not observed in wild-type
plants (Brown et al., 1999). A similar approach might also be developed for
Ca 2+ mobility in the phloem, decreasing plant susceptibility to Ca 2+ defi-
ciency disorders.
Plant treatments with growth regulators, such as ABA and GA bio-
synthesis inhibitors, have demonstrated their ability to maintain higher

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numbers of functional xylem vessels during fruit growth and development,

and can be used commercially to favor fruit Ca 2+ uptake and decrease fruit
susceptibility to Ca 2+ deficiency disorders. In addition, selection of new
fruit and leafy vegetable genotypes that maintain high numbers of func-
tional xylem vessels during growth and development can also potentially
favor higher Ca 2+ accumulation and reduce fruit and young leaf susceptibil-
ity to Ca 2+ deficiency disorders. Besides the higher number of functional
xylem vessels, the hydrostatic gradient is also required for xylem sap and
Ca 2+ movement in the fruit. In that case, techniques that favor fruit transpi-
ration, such as reduced wax deposition and shoot and mature leaf pruning,
as well as techniques that reduce mature leaf transpiration, such as reduc-
tions in WVPD, can favor fruit Ca 2+ uptake and reduce fruit susceptibility
to Ca 2+ deficiency disorders.

15.6.2  At the Cellular Level

Selection of new crop genotypes with lower Ca 2+ precipitation inside stor-
age organelles or lower Ca 2+ binding to the cell wall has been suggested to
reduce fruit susceptibility to Ca 2+ deficiency disorders. Breeding programs
could use marker-assisted selection of new genotypes that require lower
amounts of Ca 2+ for proper tissue growth and development. In addition,
treating plants with growth regulators, such as ABA and GA biosynthesis
inhibitors, has been suggested to favor an adequate cellular Ca 2+ distribu-
tion that could be used commercially to reduce fruit susceptibility to Ca 2+
deficiency disorders.

15.7  Final Considerations and Future Research Needs

More than a hundred years of study on Ca 2+ deficiency disorders has
revealed the complexity of the mechanisms involved. It is reasonable to con-
sider that many additional mechanisms remain undiscovered. For example,
although the effects of individual growth regulators have been tested on
plant susceptibility to Ca 2+ deficiency disorders, future research should
focus on growth regulator homeostasis and on the network of responses
determining fruit susceptibility to Ca 2+ deficiency disorders.
Although recent studies have shown the effect of xylem and the
xylem/phloem ratio on Ca 2+ uptake into low-transpiring sink organs, more
detailed studies are required to better understand the mechanisms regulat-
ing Ca 2+ movement in the plant. These studies may help the development
of technologies that increase the efficiency of Ca 2+ movement in plants and
into low-transpiring young leaves and fruit, which can be used to reduce
crop plant’s susceptibility to Ca 2+ deficiency disorders.

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The regulation of cellular Ca 2+ distribution has been recently sug-

gested to be the final step in the control of tissue susceptibility to Ca 2+
deficiency disorders. Therefore, better understanding of the mecha-
nisms regulating cellular Ca 2+ distribution can potentially reveal powerful
approaches to control plant tissue susceptibility to Ca 2+ deficiency disor-
ders. In addition, future studies should also better characterize the com-
bined effects of Ca 2+/ROS ratios in different cellular compartments on Ca 2+
deficiency disorders.

References
Abdal, M., and Suleiman, M. 2005. Blossom end rot occurrence in calcareous
soil of Kuwait. Acta Horticulturae 695, 63–65.
Achard, P., Gusti, A., Cheminant, S., Alioua, M., Dhondt, S., Coppens, F.,
Beemster, G.T., and Genschik, P. 2009. Gibberellin signaling controls
cell proliferation rate in Arabidopsis. Current Biology 19, 1188–1193.
Achard, P., Renou, J.P., Berthomé, R., Harberd, N.P., and Genschik, P. 2008.
Plant DELLAs restrain growth and promote survival of adversity by
reducing the levels of reactive oxygen species. Current Biology 18,
656–660.
Adams, P., and Ho, L.C. 1993. Effects of environment on the uptake and dis-
tribution of calcium in tomato and on the incidence of blossom-end rot.
Plant and Soil 154, 127–132.
Aktas, H., Karni, L., Aloni, B., and Bar-Tal, A. 2003. Physiological and bio-
chemical mechanisms leading to blossom-end rot in greenhouse-grown
peppers, irrigated with saline solution. Acta Horticulturae 609, 81–88.
Aktas, H., Karni, L., Chang, D.C., Turhan, E., Bar-Tal, A., and Aloni, B. 2005.
The suppression of salinity-associated oxygen radicals production in
pepper (Capsicum annuum) fruit by manganese, zinc and calcium in
relation to its sensitivity to blossom-end rot. Physiologia Plantarum 123,
67–74.
Aloni, R. 1987. Differentiation of vascular tissues. Annual Review of Plant
Physiology 38, 179–204.
Aloni, R. 2001. Foliar and axial aspects of vascular differentiation: Hypotheses
and evidence. Journal of Plant Growth Regulation 20, 22–34.
Amarante, C.V.T., Chaves, D.V., and Ernani, P.R. 2006a. Multivariate analy-
sis of nutritional attributes associated with bitter pit in ‘Gala’ apples.
Pesquisa Agropecuária Brasileira 41, 841–846.
Amarante, C.V.T., Chaves, D.V., and Ernani, P.R. 2006b. Mineral composi-
tion and bitter pit severity in ‘Catarina’ apples. Revista Brasileira de
Fruticultura 28, 51–54.
Amarante, C.V.T., Ernani, P.R., Blum, L.E.B., and Megguer, C.A. 2003.
Thidiazuron affects fruit set, return bloom, shoot growth, and nutrition
of apples. Pesquisa Agropecuária Brasileira 37, 1365–1372.

502
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Amarante, C.V.T., Miqueloto, A., De Freita, S.T., Steffens, C.A., Silveira, J.P.G.,
and Corrêa, T.R. 2013. Fruit sampling methods to quantify calcium and
magnesium contents to predict bitter pit development in ‘Fuji’ apple:
A multivariate approach. Scientia Horticulturae 157, 19–23.
Argenta, L.C., and Suzuki, A. 1994. Relação entre teores minerais e freqüên-
cia de “bitter pit” em maçãs cv. Gala no Brasil. Revista Brasileira de
Fruticultura 16, 267–277.
Atanasoff, D. 1934. Is bitter pit of apples a virus? Journal of Phytopathology
13, 1–8.
Bacic, A., Harris, P.J., and Stone, B.A. 1988. Structure and function of plant cell
walls. In Preiss, J. (ed.), The Biochemistry of Plants. Vol. 14. Academic
Press, New York, 297–371.
Bangerth, F. 1979. Calcium-related physiological disorders of plants. Annual
Review of Phytopathology 17, 97–122.
Banuelos, G.S., Bangerth, F., and Marschner, H. 1987. Relationship between
polar basipetal auxin transport and acropetal Ca2+ transport into tomato
fruits. Physiolgia Plantarum 71, 321–327.
Barickman, T.C., Kopsell, D.A., and Sams, C.E. 2014. Foliar applications of
abscisic acid decrease the incidence of blossom-end rot in tomato fruit.
Scientia Horticulturae 179, 356–362.
Barta, D.J., and Tibbitts, T.W. 2000. Calcium localization and tipburn develop-
ment in lettuce leaves during early enlargement. Journal of the American
Society for Horticultural Science 125, 294–298.
Batistic, O., and Kudla, J. 2010. Calcium: not just another ion. In Hell, R.,
and Mendel, R.R. (eds.), Cell Biology of Metals and Nutrients. Plant Cell
Monographs 17. Springer-Verlag, Berlin, 17–54.
Bondada, B.R., Matthews, M.A., and Shackel, K.A. 2005. Functional xylem
in the post-veraison grape berry. Journal of Experimental Botany 56,
2949–2957.
Bramlage, W.J., and Weis, S.A. 2004. Postharvest fruit quality and storage life
in relation to mineral nutrients. New York State Fruit Quarterly 12, 11–12.
Bramlage, W.J., Drake, M., and Lord, W.J. 1980. The influence of mineral
nutrition on the quality and storage performance of pome fruits grown
in North America. Acta Horticulturae 92, 29–40.
Brett, C., and Waldron, K. 1996. Physiology and Biochemistry of Plant Cell
Walls. 2nd ed. Chapman & Hall, London.
Brooks, C. 1914. Blossom-end rot of tomatoes. Phytopathology 4, 345–374.
Brown, J.W. 1926. Chemical studies in the physiology of apples. Annals of
Botany 40, 129–147.
Brown, P.H., and Hu, H. 1996. Phloem mobility of boron is species dependent:
Evidence for phloem mobility in sorbitol-rich species. Annals of Botany
77, 497–505.
Brown, P.H., Bellaloui, N., Hu, H., and Dandekar, A. 1999. Transgenically
enhanced sorbitol synthesis facilitates phloem boron transport and
increases tolerance of tobacco to boron deficiency. Plant Physiology 119,
17–20.

503
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Bush, D.S. 1995. Calcium regulation in plant cells and its role in signaling.
Annual Review of Plant Physiology and Plant Molecular Biology 46, 95–122.
Bussler, W. 1962. Ca-Mangelsymptome bei Sonnenblumen. Zeitschrift für
Pflanzenernährung, Düngung, Bodenkunde, 99, 207–215.
Bustan, A., Erner, Y., and Goldschmidt, E.E. 1995. Interaction between devel-
oping citrus fruits and their supportive vascular system. Annals of
Botany 76, 657–666.
Cakmak, I., and Römheld, V. 1977. Boron deficiency-induced impairments of
cellular functions in plants. Plant and Soil 193, 71–83.
Candan, A.P., Graell, J., and Larrigaudiere, C. 2008. Roles of climacteric eth-
ylene in the development of chilling injury in plums. Postharvest Biology
and Technology 47, 107–112.
Carne, W.M., and Martin, D. 1934. Apple investigations in Tasmania.
Miscellaneous notes, I.I. Virus theory of bitter pit. Bulletin of CSIRO
[Australia] 7, 203–214.
Carpita, N.C., and Gibeaut, D.M. 1993. Structural models of primary cell
walls in flowering plants: consistency of molecular structure with the
physical properties of the walls during growth. Plant Journal 3, 1–30.
Casado-Vela, J., Selles, S., and Martinez, R.B. 2005. Proteomic approach to
blossom-end rot in tomato fruits (Lycopersicon esculentum M.): anti-
oxidant enzymes and the pentose phosphate pathway. Proteomics 5,
2488–2496.
Chamberlain, E.E. 1933. Blossom-end rot of tomatoes. New Zealand Journal
of Agricultural Research 46, 293–296.
Chamel, A.R., and Bossy, J.P. 1981. Electron-microprobe analysis of apple
fruit tissues affected with bitter pit. Scientia Horticulturae 15, 155–163.
Chapman, E.J., and Estelle, M. 2009. Cytokinin and auxin intersection in root
meristems. Genome Biology 10, 1–5.
Chiu, T.E., and Bould, C. 1977. Sand-culture studies on the calcium nutrition
of young apple trees with particular reference to bitter pit. Journal of
Horticultural Science 52, 19–28.
Clarkson, D.T., and Hanson, J.B. 1980. The mineral nutrition of higher plants.
Annual Review of Plant Physiology 31, 239–298.
Cobb, N.A. 1895. Bitter pit of apple. Agricultural Gazette of New South Wales
6, 859–861.
Cohen, R.A., and Greene, D.W. 1989. A possible relationship between fruit
calcium and gibberellin A4 response on ‘Delicious’ apple. HortScience
24, 223.
Collier, G.F., and Tibbitts, T.W. 1982. Tipburn of lettuce. Horticultural Reviews
4, 49–65.
Conn, S.J., Gilliham, M., Athman, A., Schreiber, A.W., Baumann, U., Moller, I.,
Cheng, N.H., Stancombe, M.A., Hirschi, K.D., Webb, A.A.R., Burton, R.,
Kaiser, B.N., Tyerman, S.D., and Leigh, R.A. 2011. Cell-specific vacuolar
calcium storage mediated by CAX1 regulates apoplastic calcium con-
centration, gas exchange, and plant productivity in Arabidopsis. Plant
Cell 23, 240–257.

504
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Cutting, J.G.M., and Bower, J.P. 1989. The relationship between basipetal
auxin transport and calcium allocation in vegetative and reproductive
flushes in avocado. Scientia Horticulturae 41, 27–34.
Dekock, P.C., Vaughan, D., Hall, A., and Ord, B.G. 1980. Biochemical studies
on blossom-end rot of tomatoes. Physiologia Plantarum 48, 312–316.
Dela Fuente, R.K., Tang, P.M., and Guzman, C.C. 1986. The requirement
of  calcium and boron in auxin transport. In Plant Growth Substances,
Bopp, M. (ed.), Springer-Verlag, Berlin, 227–230
Demarty, M., Morvan, C., and Thellier, M. 1984. Calcium and the cell wall.
Plant, Cell and Environment 7, 441–448.
Dewitte, W., and Murray, J.A. 2003. The plant cell cycle. Annual Review of
Plant Biology 54, 235–264.
Dickinson, D.B. 1978. Influence of borate and pentaerythritol concentrations
on germination and tube growth of Lolium longiflorum pollen. Journal of
the American Society for Horticultural Science 103, 414–416.
Dickson, B., Sagar, G.R., and Shorrocks, V.M. 1973. Effect of calcium and
boron on the incidence of tree and storage pit in apples of the cultivar
Egrement Russet. Journal of Horticultural Science 48, 403–411.
Dolmetsch, R.E., Lewis, R.S., Goodnow, C.C., and Healy, J.I. 1997. Differential
activation of transcription factors by Ca2+ response amplitude and dura-
tion. Nature 386, 855–858.
Dražeta, L., Lang, A., Cappellini, C., Hall, A.J., Volz, R.K., and Jameson, P.E.
2004b. Vessel differentiation in the pedicel of apple and the effects of
auxin transport inhibition. Physiologia Plantarum 120, 162–170.
Dražeta, L., Lang, A., Hall, A.J., Volz, R.K., and Jameson, P.E. 2004a. Causes
and effects of changes in xylem functionality in apple fruit. Annals of
Botany 93, 275–282.
Dris, R., Niskanen, R., and Fallahi, E. 1998. Nitrogen and calcium nutrition
and fruit quality of commercial apple cultivars grown in Finland. Journal
of Plant Nutrition 21, 2389–2402.
Ehrhardt, R.J., Wais, R., and Long, S.R. 1996. Calcium spiking in plant root
hairs responding to rhizobium nodulation signals. Cell 85, 673–681.
Elfving, D.C., and Cline, R.A. 1993. Cytokinin and ethephon affects crop
load, shoot growth, and nutrient concentration of ‘Empire’ apple trees.
HortScience 28, 1011–1014.
Fallahi, E., Fallahi, B., and Peryea, F.J. 2010. Effects of mineral nutrition
on fruit quality and nutritional disorders in apples. Acta Horticulturae
868, 49–59.
Faust, M., and Shear, C.B. 1968. Corking disorders of apples: A physiological
and biochemical review. Botanical Review 34, 441–469.
Francois, L.E., Donovan, T.J., and Maas, E.V. 1991. Calcium deficiency of arti-
choke buds in relation to salinity. HortScience 26, 549–553.
Ferguson, I.B., and Watkins, C.B. 1989. Bitter pit in apple fruit. Horticultural
Reviews 11, 289–355.
Freitas, S.T., and Mitcham, E.J. 2012. Factors involved in fruit calcium defi-
ciency disorders. Horticultural Reviews 40, 107–146.

505
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Freitas, S.T., Amarante, C.V.T., Dandekar, A.M., and Mitcham, E.J. 2013. Shading
affects flesh calcium uptake and concentration, bitter pit incidence and
other fruit traits in ‘Greensleeves’ apple. Scientia Horticulturae 161, 266–272.
Freitas, S.T., Amarante, C.V.T., Labavitch, J.M., and Mitcham, E.J. 2010.
Cellular approach to understand bitter pit development in apple fruit.
Postharvest Biology and Technology 57, 6–13.
Freitas, S.T., Handa, A.K., Wu, Q., Park, S., and Mitcham, E.J. 2012b. Role of
pectin methylesterase in cellular calcium distribution and blossom-end
rot development in tomato fruit. Plant Journal 71, 824–835.
Freitas, S.T., Jiang, C.Z., and Mitcham, E.J. 2012a. Mechanisms involved in
calcium deficiency development in tomato fruit in response to gibberel-
lins. Journal of Plant Growth Regulation 31, 221–234.
Freitas, S.T., McElrone, A.J., Shackel, K.A., and Mitcham, E.J. 2014. Calcium
partitioning and allocation and blossom-end rot development in tomato
plants in response to whole-plant and fruit-specific abscisic acid treat-
ments. Journal of Experimental Botany 65, 235–247.
Freitas, S.T., Padda, M., Wu, Q., Park, S., and Mitcham, E.J. 2011a. Dynamic
alterations in cellular and molecular components during blossom-end
rot development in tomatoes expressing sCAX1, a constitutively active
Ca2+/H+ antiporter from Arabidopsis. Plant Physiology 156, 844–855.
Freitas, S.T., Shackel, K.A., and Mitcham, E.J. 2011b. Abscisic acid triggers
whole-plant and fruit-specific mechanisms to increase fruit calcium
uptake and prevent blossom end rot development in tomato fruit.
Journal of Experimental Botany 62, 2645–2656.
Fuller, M.M. 1980. Cell ultra-structure in apple fruits in relation to calcium
concentration and fruit quality. Acta Horticulturae 92, 51–55.
Galloway, B.T. 1888. Notes on the black-rot of tomatoes. Report of the
Commissioner of Agriculture. U.S. Department of Agriculture,
Washington, DC, 339–345.
Gillaspy, G., Ben-David, H., and Gruissem, W. 1993. Fruits: A developmental
perspective. Plant Cell 5, 1439–1451.
Granelli, G., and Ughini, V. 1990. Leaf boron contents and bitter pit in apple.
Acta Horticulturae 274, 169–174.
Greene, D.W. 1993. A review of the use of benzyladenine (BA) as a chemical
thinner for apples. Acta Horticulturae 329, 231–236.
Greene, D.W. 1995. Thidiazuron effects on fruit set, fruit quality, and return
bloom of apples. HortScience 30, 1238–1240.
Greene, D.W., Autio, W.R., and Miller, P. 1990. Thinning activity of benzylad-
enine on several apple cultivars. Journal of the American Society for
Horticultural Science 115, 394–400.
Guichard, S., Gary, C., Leonardi, C., and Bertin, N. 2005. Analysis of growth
and water relations of tomato fruit in relation to air vapor pressure defi-
cit and plant fruit load. Journal of Plant Growth Regulation 24, 201–213.
Hammouda, A.M. 1987. Blossom-end rot of watermelon in the southern
region of Oman (Dhofan). Journal of Agricultural Science 108, 667–669.

506
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Hanson, J.B. 1960. Impairment of respiration, ion accumulation, and ion

retention in root tissue treated with ribonuclease and ethylenediamine
tetraacetic acid. Plant Physiology 35, 372–379.
Hauser, H., Levine, B.A., and Williams, R.J.P. 1976. Interaction of ions with
membranes. Trends in Biochemical Sciences 1, 278–281.
Hepler, P.K., and Wayne, R.O. 1985. Calcium and plant development. Annual
Review of Plant Physiology 36, 397–439.
Hewitt, E.J. 1963. The essential nutrient elements: Requirements and interac-
tions in plants. In Steward, F.C. (ed.), Plant Physiology. Vol. 3. Academic
Press, New York, 137–329.
Hirschi, K.D. 2004. The calcium conundrum. Both versatile nutrient and spe-
cific signal. Plant Physiology 136, 2438–2442.
Ho, L.C., and White, P.J. 2005. A cellular hypothesis for the induction of blos-
som-end rot in tomato fruit. Annals of Botany 95, 571–581.
Ho, L.C., Belda, R., Brown, M., Andrews, J., and Adams, P. 1993. Uptake
and transport of calcium and the possible causes of blossom-end rot in
tomato. Journal of Experimental Botany 44, 509–518.
Hochmuth, G.J. and Hochmuth, R.C. 2012. Blossom-end rot in bell pep-
per: causes and prevention. Copyright@2015 University of Florida,
Institute of Food and Agricultural Sciences, http://edis.ifas.ufl.edu/
ss497#FIGURE%203 (accessed June 16th, 2015).
Islam, A.K.M.S., Asher, C.J., and Edwards, D.G. 1987. Response of plants to
calcium concentration in flowing solution culture with chloride or sul-
phate as the counter-ion. Plant and Soil 98, 377–395.
Jager, G. 1869. Uber das pelzig oder stippigwerden der kernobstfrucht. Illustr.
Monatshefte fur Obst- u Weinbau 318–319.
Johnson, C.H., Knight, M.R., Kondo, T., Masson, P., Sedbrook, J., Haley, A.,
and Trewavas, A.J. 1995. Circadian oscillations of cytosolic and chloro-
plastic free calcium in plants. Science 269, 1863–1865.
Jones, R.L., and Bush, D.S. 1991. Gibberellic acid regulates the level of a BiP
cognate in the endoplasmic reticulum. Plant Physiology 97, 456–459.
Jaiswal, J.K. 2001. Calcium—How and why? Journal of Biosciences 26, 357–363.
Karley, A.J., and White, P.J. 2009. Moving cationic minerals to edible tissues:
Potassium, magnesium, calcium. Current Opinion in Plant Biology 12,
291–298.
Kauss, H. 1987. Some aspects of calcium-dependent regulation in plant
metabolism. Annual Review of Plant Physiology 38, 47–72.
Kendall, J.M., Dormer, R.L., and Campbell, A.K. 1992. Targeting aequorin to
the endoplasmic reticulum of living cells. Biochemical and Biophysical
Research Communications 189, 1008–1016.
Kirby, E.A., and Pilbeam, D.J. 1984. Calcium as a plant nutrient. Plant, Cell
and Environment 7, 397–405.
Knight, H., Trewavas, A.J., and Knight, M.R. 1996. Cold calcium signaling in
Arabidopsis thaliana involves two cellular pools and a change in calcium
signature after acclimation. Plant Cell 8, 489–503.

507
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Koike, S. and Smith, R. 2010. Calcium deficiency disorders hit vegetable

crops in central coast. Copyright©2015 Regents of the University of
California, Agriculture and Natural Resources, http://ucanr.org/blogs/
blogcore/postdetail.cfm?postnum=3407 (accessed June 16th, 2015).
Kwak, J.M., Nguyen, V., and Schroeder, J.I. 2006. The role of reactive oxygen
species in hormonal responses. Plant Physiology 141, 323–329.
Lanauskas, J., and Kvikliene, N. 2006. Effect of calcium foliar application on
some fruit quality characteristics of ‘Sinap Orlovskij’ apple. Agronomy
Research 4, 31–36.
Lang, A. 1990. Xylem, phloem and transpiration flows in developing apple
fruits. Journal of Experimental Botany 41, 645–651.
Legge, R.L., Thompson, J.E., Baker, J.E., and Lieberman, M. 1982. The effect
of calcium on the fluidity of phase properties of microsomal membranes
isolated from postclimacteric Golden Delicious apples. Plant and Cell
Physiology 23, 161–169.
Li, J., Wang, X.Q., Watson, M.B., and Assmann, S.M. 2000. Regulation of
abscisic acid-induced stomatal closure and anion channels by guard cell
AAPK kinase. Science 287, 300–303.
Liu, Y., Zhao, Z., Si, J., Di, C., Han, J., and An, L. 2009. Brassinosteroids
alleviate chilling-induced oxidative damage by enhancing antioxidant
defense system in suspension cultured cells of Chorispora bungeana.
Plant Growth Regulation 59, 207–214.
Logan, D.C., and Knight, M.R. 2003. Mitochondrial and cytosolic calcium
dynamics are differentially regulated in plants. Plant Physiology 133,
21–24.
Lötze, E., and Theron, K.I. 2006. Existing pre-harvest predictions and models
for bitter pit incidence. South African Fruit Journal 5, 20–25.
Lötze, E., Theron, K.I., and Joubert, J. 2010. Assessment of pre-harvest
physiological infiltration methods for predicting commercial bit-
ter pit in ‘Braeburn’ and ‘Golden Delicious’. Acta Horticulturae 868,
347–352.
Ludders, P. 1979. The effect of nitrogen nutrition on bitter pit in apples.
Communications in Soil Science and Plant Analysis 10, 401–415.
Lund, Z.F. 1970. The effect of calcium and its relation to several cations in soy-
bean root growth. Soil Science Society of America Journal 34, 456–459.
MacArthur, M. 1940. Histology of some physiological disorders of the apple
fruit. Canadian Journal of Research 18, 26–39.
Marcelis, L.F.M., and Ho, L.C. 1999. Blossom-end rot in relation to growth rate
and calcium content in fruits of sweet pepper (Capsicum annuum L.).
Journal of Experimental Botany 50, 357–363.
Marschner, H. 1995. Mineral Nutrition of Higher Plants. 2nd ed. Academic
Press, London.
Maynard, D.N., and Hopkins, D.L. 1999. Watermelon fruit disorders.
HortTechnology 9, 155–161.
Maynard, D.N., Warner, D.C., and Howell, J.C. 1981. Cauliflower leaf tipburn:
A calcium deficiency disorder. HortScience 16, 193–195.

508
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

McAinsh, M.R., Webb, A.A.R., Taylor, J.E., and Hetherington, A.M. 1995.
Stimulus-induced oscillations in guard-cell cytosolic free calcium. Plant
Cell 7, 1207–1219.
Mestre, T.C., Garcia-Sanchez, F., Rubio, F., Martinez, V., and Rivero, R.M.
2012. Glutathione homeostasis as an important and novel factor con-
trolling blossom-end rot development in calcium-deficient tomato fruits.
Journal of Plant Physiology 169, 1719–1727.
Mielke, E.A., and Chaplin, M.H. 2008. Controlling post-harvest storage problems
on ‘Concorde’ pear with boron application. Acta Horticulturae 800, 571–575.
Monge, E., Aguirre, R., and Blanco, A. 1994. Application of paclobutrazol and
GA3 to adult peach trees: effects on nutritional status and photosyn-
thetic pigments. Journal of Plant Growth Regulation 13, 15–19.
Mulder, D. 1952. Nutritional studies on fruit trees. Plant and Soil 4, 107–117.
Nachtigall, G.R., and Freire, C.J.S. 1998. Previsão da incidência de “bitter
pit” em maçãs através dos teores de cálcio em folhas e frutos. Revista
Brasileira de Fruticultura 20, 158–166.
Neilsen, G.H., Neilsen, D., Toivonen, P., and Herbert, L. 2008. Annual bloom-
time phosphorus fertigation affects soil phosphorus, apple tree phos-
phorus nutrition, yield, and fruit quality. HortScience 43, 885–890.
Nonami, H., Fukuyama, T., Yamamoto, M., Yang, L., and Hashimoto, Y. 1995.
Blossom-end rot of tomato plants may not be directly caused by calcium
deficiency. Acta Horticulturae 396, 107–114.
Oberly, G.H., and Kenworthy, A.L. 1961. Effect of mineral nutrition on the
occurrence of bitter pit in Northern Spy apples. Proceedings of the
American Society for Horticultural Science 77, 29–34.
Olle, M., and Bender, I. 2009. Causes and control of calcium deficiency dis-
orders in vegetables: A review. Journal of Horticultural Science and
Biotechnology 84, 577–584.
O’Neill, M., Albersheim, P., and Darvill, A. 1990. The pectic polysaccharides
of primary cell walls. In Dey, P.M. (ed.), Methods in Plant Biochemistry,
Vol. 2. Academic Press, London, 415–441.
Ontario Crop Integrated Pest Management (IPM). 2009. Blossom-end rot.
Copyright © Queen’s Printer for Ontario, 2015. http://www.omafra.gov.
on.ca/IPM/english/cucurbits/diseases-and-disorders/blossom-end-
rot.html (accessed June 16th, 2015).
Park, S., Cheng, N.H., Pittman, J.K., Yoo, K.S., Park, J., Smith, R.H., and
Hirschi, K.D. 2005. Increasing calcium levels and prolonged shelf life in
tomatoes expressing Arabidopsis H+/Ca2+ transporters. Plant Physiology
139, 1194–1206.
Perrot-Rechenmann, C. 2010. Cellular responses to auxin: Division versus
expansion. Cold Spring Harbor Perspectives in Biology 2, 1–15.
Picchioni, G.A., Watada, A.E., Conway, W.S., Whitaker, B.D., and Sams, C.E.
1998. Postharvest calcium infiltration delays membrane lipid catabolism
in apple fruit. Journal of Agricultural and Food Chemistry 46, 2452–2457.
Plieth, C. 2005. Calcium: just another regulator in the machinery of life?
Annals of Botany 96, 1–8.

509
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Qiu, Y., Xi, J., Du, L., and Poovaiah, B.W. 2012. The function of calreticulin
in plant immunity—New discoveries for an old protein. Plant Signaling
and Behavior 7, 907–910.
Raese, J.T., and Drake, S.R. 1997. Nitrogen fertilization and elemental compo-
sition affects fruit quality of ‘Fuji’ apples. Journal of Plant Nutrition 20,
1797–1809.
Raleigh, S.M., and Chucka, J.A. 1944. Effect of nutrient ratio and concentra-
tion on growth and composition of tomato plants and on the occurrence
of blossom-end rot of the fruit. Plant Physiology 19, 671–678.
Ridley, B.L., O’Neill, M.A., and Mohnen, D. 2001. Pectins: Structure, biosynthe-
sis, and oligogalacturonide-related signaling. Phytochemistry 57, 929–967.
Rosen, C.J. 1990. Leaf tipburn in cauliflower as affected by cultivar, calcium
sprays, and nitrogen nutrition. HortScience 25, 660–663.
Rudd, J.J., and Franklin-Tong, V.E. 1999. Calcium signaling in plants. Cellular
and Molecular Life Sciences 55, 214–232.
Rudell, D.R., Fellman, J.K., and Mattheis, J.P. 2005. Preharvest application of
methyl jasmonate to ‘Fuji’ apples enhances red coloration and affects
fruit size, splitting, and bitter pit incidence. HortScience 40, 1760–1762.
Rudell, D.R., Mattheis, J.P., Fan, X., and Fellman, J.K. 2002. Methyl jasmonate
enhances anthocyanin accumulation and modifies production of phe-
nolics and pigments in ‘Fuji’ apples. Journal of the American Society for
Horticultural Science 127, 435–441.
Saure, M.C. 1996. Reassessment of the role of calcium in development of bit-
ter pit in apple. Australian Journal of Plant Physiology 23, 237–243.
Saure, M.C. 1998. Causes of tipburn disorder in leaves of vegetables. Scientia
Horticulturae 76, 131–147.
Saure, M.C. 2001. Blossom-end rot of tomato (Lycopersicon esculentum Mill.): a
calcium or a stress-related disorder? Scientia Horticulturae 90, 193–208.
Saure, M.C. 2005. Calcium translocation to fleshy fruit: Its mechanism and
endogenous control. Scientia Horticulturae 105 65–89.
Saure, M.C. 2014. Why calcium deficiency is not the cause of blossom-end rot in
tomato and pepper fruit—A reappraisal. Scientia Horticulturae 174, 151–154.
Schenabel, H., Roth, U., and Friebe, A. 2001. Brassinosteroids-induced stress
tolerances of plants. Recent Research Developments in Phytochemistry 5,
169–183.
Schonherr, J., and Bukovac, M.J. 1973. Ion exchange properties of isolated
tomato fruit cuticular membrane: exchange capacity, nature of fixed
charges and cation selectivity. Planta 109, 73–93.
Scrase-Field, S.A.M.G., and Knight, M.R. 2003. Calcium: Just a chemical
switch? Current Opinion in Plant Biology 6, 500–506.
Sen, F., Karacali, I., Irget, M.E., Elmaci, O.L., and Tepecik, M. 2010. A new strat-
egy to enrich calcium nutrition of fruit: Synergic effects of postharvest
foliar calcium and boron sprays. Journal of Plant Nutrition 33, 175–184.
Sharples, R.O. 1964. The effects of fruit thinning on the development of Cox’s
Orange Pippin apples in relation to the incidence of storage disorders.
Journal of Horticultural Science 39, 224–235.

510
 C alci um D e f i c i e n cy D iso rd ers in Pl a nts

Shear, C.B. 1975. Calcium-related disorders of fruit and vegetables.

HortScience 10, 361–365.
Shear, C.B., and Faust, M. 1971. Don’t neglect calcium in your apple tree’s
diet. American Fruit Grower Magazine 91, 18–20, 23.
Simon, E.W. 1978. The symptoms of calcium deficiency in plants. New
Phytologist 80, 1–15.
Smith, A.J.M. 1926. Bitter pit in apples: A review of the problem. Special
Report 28. Food Investigation Board, London, UK, 23.
Sorensen, I., Domozych, D., and Willats, W.G.T. 2010. How have plant cell
walls evolved? Plant Physiology 153, 366–372.
Steenkamp, J., and de Villiers, O.T. 1979. The effect of growth regulators
on the uptake and distribution of calcium in Golden Delicious apples.
Agroplantae 11, 79–81.
Steveninck, R.F.M.V. 1965. The significance of calcium on the apparent per-
meability of cell membranes and the effects of substitution with other
divalent ions. Physiologia Plantarum 18, 54–69.
Stevens, J., Senaratna, T., and Sivasithamparam, K. 2006. Salicylic acid
induces salinity tolerance in tomato (Lycopersicon esculentum cv. Roma):
associated changes in gas exchange, water relations and membrane sta-
bilization. Plant Growth Regulation 49, 77–83.
Suzuki, K., Shono, M., and Egawa, Y. 2003. Localization of calcium in the
pericarp cells of tomato fruit during the development of blossom-end
rot. Protoplasma 222, 149–156.
Suzuki, K., Takeda, H., and Egawa, Y. 2000. Morphological aspect of blossom-
end rot fruits of tomato. Acta Horticulturae 511, 257–264.
Tadesse, T., Nichols, M.A., Hewett, E.W., and Fisher, K.J. 2001. Relative
humidity around the fruit influences the mineral composition and
incidence of blossom-end rot in sweet pepper. Journal of Horticultural
Science and Biotechnology 76, 9–16.
Taylor, M.D., and Locascio, S.J. 2004. Blossom-end rot: A calcium deficiency.
Journal of Plant Nutrition 27, 123–139.
Turhan, E., Karni, L., Aktas, H., Deventurero, G., Chang, D.C., Bar-Tal,
A., and Aloni, B. 2006. Apoplastic antioxidants in pepper (Capsicum
annuum L.) fruit and their relationship to blossom-end rot. Journal of
Horticultural Science and Biotechnology 81, 661–667.
Ubeda-Tomás, S., Beemster, G.T.S., and Bennett, M.J. 2012. Hormonal regu-
lation of root growth: Integrating local activities into global behaviour.
Trends in Plant Science 17, 326–331.
Vang-Petersen, O. 1980. Calcium nutrition of apple trees: A review. Scientia
Horticulturae 12, 1–9.
van Schreven, A.C. 1961. Bitter pit. Bulletin of the International Institute of
Refrigeration 1, 167–171.
Xuan, H., Streif, J., Pfeffer, H., Dannel, F., Römheld, V., and Bangerth, F. 2001.
Effects of pre-harvest boron application on the incidence of CA-storage
related disorders in ‘Conference’ pears. Journal of Horticultural Science
and Biotechnology 76, 133–137.

511
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Xuan, H., Streif, J., Saquet, A., Römheld, V., and Bangerth, F. 2005. Application
of boron with calcium affects respiration and ATP/ADP ratio in
‘Conference’ pears during controlled atmosphere storage. Journal of
Horticultural Science and Biotechnology 80, 633–637.
Wallace, A., Frohlich, E., and Lunt, O.R. 1966. Calcium requirements of
higher plants. Nature 209, 634.
Wang, H., Qi, Q., Schorr, P., Cutler, A.J., Crosby, W.L., and Fowke, L.C. 1998.
ICK1, a cyclin-dependent protein kinase inhibitor from Arabidopsis
­thaliana interacts with both Cdc2a and CycD3, and its expression is
induced by abscisic acid. Plant Journal 15, 501–510.
Wedgworth, H.H., Neal, D.C., and Wallace, J.M. 1926. Wilt and blossom-
end rot of the tomato. Bulletin No. 247. Raymond Branch Experiment
Station, Raymond, MS.
White, P.J. 2001. The pathways of calcium movement to the xylem. Journal of
Experimental Botany 52, 891–899.
White, P.J., and Broadley, M.R. 2003. Calcium in plants. Annals of Botany 92,
487–511.
Williams, R.J.P. 1970. The biochemistry of sodium, potassium, magnesium,
and calcium. Quarterly Review of the Chemical Society 24, 331–365.
Wojcik, P., and Treder, W. 2006. Effect of drip boron fertigation on yield and
fruit quality in a high-density apple orchard. Journal of Plant Nutrition
29, 2199–2213.
Wojcik, P., and Wojcik, M. 2003. Effects of boron fertilization on ‘Conference’
pear tree vigor, nutrition, and fruit yield and storability. Plant and Soil
256, 413–421.
Wojcik, P., Cieslinski, G., and Mika, A. 1999. Apple yield and fruit qual-
ity as influenced by boron applications. Journal of Plant Nutrition 22,
1365–1377.
Wojcik, P., Wojcik, M., and Klamkowski, K. 2008. Response of apple trees
to boron fertilization under conditions of low soil boron availability.
Scientia Horticulturae 116, 58–64.
Wu, Q., Shigaki, T., Han, J.S., Kim, C.K., Hirschi, K.D., and Park, S. 2012.
Ectopic expression of a maize calreticulin mitigates calcium deficiency-
like disorders in sCAX1-expressing tobacco and tomato. Plant Molecular
Biology 80, 609–619.
Yang, H., Jie, Y., Li, J., Zhang, W., and Fan, W. 2008. The regulation of bioregu-
lator to calcium absorption and its kinetics in apple rootstocks roots.
Acta Horticulturae 774, 259–264.
Yang, H., Jie, Y., Zhang, L.Z., and Cui, M.G. 2004. The effect of IBA on the
Ca2+ absorption and Ca2+-ATPase activity and their ultracytochemical
localization in apple roots. Acta Horticulturae 636, 211–219.
Yermiyahu, U., Nir, S., Ben-Hayyim, G., and Kafkafi, U. 1994. Quantitative
competition of calcium with sodium or magnesium for sorption sites
on plasma membrane vesicles of melon (Cucumis melo L.) root cells.
Journal of Membrane Biology 138, 55–63.

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Chapter 16

Fresh Fruit Aroma:

An Integrative
Overview for a
Complex Flavor Trait
Orianne Gudenschwager and Bruno G. Defilippi
Instituto de Investigaciones Agropecuarias
(CRI La Platina), La Pintana, Santiago, Chile

Abstract 514
16.1 Introduction 515
16.2 Aroma Composition in Fruits 516
16.2.1 Apple 516
16.2.2 Melon 517
16.2.3 Strawberry 517
16.2.4 Tomato 518
16.2.5 Citrus 518
16.2.6 Grape 519
16.2.7 Peach 519
16.2.8 Banana 520

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16.3 Biosynthesis and Regulation of Aroma Volatiles in Fruit 520

16.3.1  Biosynthetic Pathways of Aroma Volatiles 520
16.3.1.1 Fatty Acid Metabolism 520
16.3.1.2 Amino Acid Metabolism 522
16.3.1.3 Ester Biosynthesis 522
16.3.1.4 Carbohydrate Metabolism 524
16.3.2  Aroma Modulation during Fruit Ripening 525
16.4 Influence of Pre- and Postharvest Factors on
Fruit Aroma 527
16.4.1  Preharvest Factors 527
16.4.1.1 Genotype 527
16.4.1.2 Growing Conditions 529
16.4.1.3 Fruit Maturity 531
16.4.2  Postharvest Technologies 532
16.4.2.1 Storage Temperature 532
16.4.2.2 Storage Atmosphere 534
16.4.2.3 Ethylene Control 536
16.4.2.4 Other Technologies 537
16.5 Conclusions and Future Perspectives 538
Acknowledgments 539
References 539

Abstract
Fruit consumers are not only looking for traditional quality attributes such
as sugar, acidity, firmness, and appearance. They also value other attri-
butes, including nutrient availability, antioxidants, and aroma, where each
of these attributes is the result of a complex interaction between intrinsic
and external variables.
Overall aroma is determined by a complex mixture of a large number
of odor-active volatile compounds, and differences in the concentrations
and thresholds of key volatiles ultimately determine the distinctive aroma
of a particular fruit species or cultivar. It is also the result of a dynamic
process, where during fruit development, there are many changes of
these m ­ etabolites caused by their synthesis, transport, or degradation.
Finally, there are several pre- and postharvest factors affecting tree and
fruit p
­ hysiology, leading to important changes in fruit composition, such as
aroma-related volatiles.
Several studies have been performed identifying and characterizing
the most important genes and encoded enzymes involved in aroma-related
volatiles; however, research in the mechanisms of regulation or modulation

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is still limited. In this chapter, we provide an overview of the most impor-

tant metabolic pathways and genes that control volatile biosynthesis in
model fruits, including apple, melon, strawberry, and tomato, with a special
emphasis on fruit ripening and the role of ethylene during this ­process.
Also presented are a brief description of the composition of volatiles in vari-
ous fruit species and a discussion of the influences of preharvest factors
and postharvest technologies on fruit aroma.

16.1 Introduction
The quality of fresh fruit is related to many attributes, such as appearance,
texture, color, and flavor, with the last relying on a combination of taste
and aroma. In addition to its biological importance to fruit species survival,
aroma is a key contributor to fruit flavor perception and plays a primary role
in consumer acceptability.
A complex mixture of a large number of odor-active volatile compounds
determines the aroma characteristics (Dixon and Hewett, 2000), where dif-
ferences in the concentrations and thresholds of key volatiles ultimately
determine the distinctive aroma of a particular fruit species or cultivar
(Defilippi et al., 2009). With the advent of highly sensitive instrumental meth-
ods, a large number of volatile compounds have been identified in fruits, and
in combination with sensory analysis, these methods have made it possible
to categorize volatile compounds according to their organoleptic importance.
Research on fruit aroma has revealed that volatile compounds are made up
of diverse classes of chemical groups, including acids, aldehydes, ketones,
alcohols, esters, sulfur compounds, furans, phenols, terpenes, epoxides,
and lactones (Schwab et al., 2008), all of which are derived from different
biosynthetic pathways that are highly integrated with fruit development and
ripening. Volatile compounds are generally produced during ripening via the
metabolism of high-molecular-weight molecules such as proteins, carbohy-
drates, and fatty acids (Aharoni and Lewinsohn, 2010), processes that are
considered to be mediated by ethylene in climacteric fruit.
Significant advances have been made in recent years toward improv-
ing our knowledge of the biosynthetic pathways and regulation of aroma-
related volatiles. These advances have resulted in a better understanding
of the mechanisms involved in aroma production, as well as the influ-
ences of environmental factors, genetic background, and handling
procedures on fruit aroma. However, there are still many underlying
mechanisms of aroma volatile biosynthesis and modulation that remain
to be determined.
This chapter provides an overview of the critical metabolic pathways
and genes that control volatile biosynthesis in model fruits, with a special
emphasis on fruit ripening and the role of ethylene during this process.

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Also presented are a brief description of the composition of volatiles in

­various fruit species and a discussion of the influences of preharvest f­ actors
and postharvest technologies on fruit aroma.

16.2  Aroma Composition in Fruits

In recent years, instrumental analysis with gas chromatography (GC) cou-
pled with mass spectroscopy has made it possible to identify a broad range
of volatile compounds in fresh fruit. These types of analysis, combined with
sensory analyses, have provided great insights into the impact of volatile
compounds on the perception of flavor by consumers. The characterization
of fruit volatiles has revealed that the major chemical groups that appear to
be common to several fruits are alcohols, aldehydes, and esters (Defilippi
et al., 2009). Volatile esters are the most significant contributors to aroma in
several fruits, such as apples, bananas, melons, and strawberries. Volatile
esters give all of these fruits their characteristic fruity flavor, whereas
­a lcohols and aldehydes contribute to fruit aroma and also serve as precur-
sors for ester biosynthesis (Song and Forney, 2008).
As volatile compounds can be synthesized either in intact fruit
­tissue prior to consumption or after tissue disruption caused by biting
and mastication (Contreras and Beaudry, 2013), section 16.3 reviews
the major volatile compounds, which in some cases are synthesized by
these two routes, that contribute to the aroma of some climacteric and
nonclimacteric fruits. These fruits were selected on the basis of their eco-
nomic importance and the amount of relevant information published in
the literature.

16.2.1 Apple
More than 350 volatile compounds, such as esters, alcohols, aldehydes,
ketones, terpenes, and ethers, have been reported as contributing to
the aroma profile of apples (Zhu et al., 2008; Dunemann et al., 2012). Of
these, esters, aldehydes, and alcohols are considered the most significant
contributors to the typical apple aroma (Plotto et al., 2000; Contreras and
Beaudry, 2013). However, which compounds are most important depends
on the developmental and ripening stage of the fruit. Esters are the most
abundant volatiles in apples, accounting for up to 80%–98% of total volatiles
(Altisent et al., 2009). Both straight- and branched-chain esters are found in
apples (Song and Forney, 2008), and among these, butyl acetate, 2-methyl
butyl acetate, and hexyl acetate are considered the major contributors to
the characteristic “red apple aroma” and “Cox-like aroma” in most apple

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cultivars (Holland et al., 2005; Zhu et al., 2005; Dunemann et al., 2009). The
alcohols 2-methyl propanol, (Z)-hexen-3-ol, 1-hexanol, and 1-octanol and
the aldehyde 1-hexanal are considered key volatile compounds in cultivars
such as ‘Golden Delicious’ and ‘Fuji’ (Altisent et al., 2009).

16.2.2 Melon
Of the approximately 240 volatile compounds identified in various melon
varieties (Shan et al., 2012), straight- and branched-chain esters, satu-
rated and unsaturated aldehydes, alcohols, and sulfur compounds are
likely to be the major contributors to the aroma of melon (Obando-Ulloa
et al., 2008; Verzera et al., 2011). As melon fruit contains climacteric and
nonclimacteric genotypes, the aroma profile depends on the ripening
behavior. Climacteric varieties, which are considered highly aromatic,
are composed predominantly of esters (Shalit et al., 2001), mainly in the
form of acetate derivatives, such as methyl butyl, ethyl, hexyl, nonenyl,
and benzyl acetates (Song and Forney, 2008). Among these, ethyl butano-
ate, methyl-2-methyl butanoate, and ethyl-2-methyl propanoate contribute
to the fruity and sweet aroma notes of muskmelon (Condurso et al., 2012).
In contrast, nonclimacteric varieties are normally less aromatic due to
the lack of volatile esters and their high levels of aldehydes and alcohols
(Gonda et al., 2010).

16.2.3 Strawberry
Strawberries possess one of the most complex fruit aromas, and therefore
their typical aroma has been extensively studied. Approximately 360 vola-
tile compounds contribute to the aroma of strawberries (Zorrilla-Fontanesi
et al., 2012), including esters, aldehydes, ketones, alcohols, terpenes, fura-
nones, and sulfur compounds (Vandendriessche et al., 2013). Straight esters
and furanones appear to be the most important aroma-active compounds
in strawberries (Blanch et al., 2011), with the esters being represented by
ethyl hexanoate, ethyl butanoate, methyl butanoate, and methyl hexanoate
(Jouquand and Chandler, 2008; Almenar et al., 2009), and the furanones pri-
marily represented by furaneol (2,5-dimethyl-4-hydroxy-3(2H)-­furanone)
and mesifurane (2,5-dimethyl-4-methoxy-3(2H)-furanone). Esters contrib-
ute green and sweet fruity notes (Ménager et al., 2004), while furanones
contribute sweet caramel-like and floral notes (Jouquand and Chandler,
2008). Other compounds that contribute to strawberry aroma are terpenes
(linalool, nerolidol, terpineol, and α-pinene), which provide citrus and
spicy notes (Zorrilla-Fontanesi et al., 2012), and aldehydes (hexanal and

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2-hexenal) and alcohols, such as 3-hexenol, which contribute green and

unripe notes to strawberry aroma (Vandendriessche et al., 2013).

16.2.4 Tomato
More than 400 aroma volatiles have been identified in intact fruit and
upon tissue disruption (Díaz de León-Sánchez et al., 2009; Bai et al., 2011).
These volatiles include hydrocarbons, alcohols, phenols, ethers, aldehydes,
ketones, carboxylic acids, esters, and lactones, among others (Zhu et al.,
2005). A combination of cis-3-hexenal, cis-3-hexenol, hexanal, 1-penten-
3-one, 3-ethyl butanal, trans-2-hexenal, 6-methyl-5-hepten-2-one, methyl
salicylate, 2-isobutylthiazole, and β-ionone, at appropriate concentrations,
provides the characteristic fresh ripe aroma of tomatoes (Baldwin et al.,
2000; Bai et al., 2011). Another important volatile compound that influences
flavor is the monoterpene S-linalool, which imparts a sweet, floral, alcoholic
note (Lewinsohn et al., 2001).

16.2.5 Citrus
The citrus aroma has been difficult to study because citrus juices are
­l iquid–solid suspensions, and suspended solids can alter the headspace
concentrations and aroma thresholds of many volatiles (Rouseff et al.,
2009). There are three sources from which citrus volatiles are gener-
ated: the juice contained in the juice sacs, which provide volatile water-­
soluble compounds; juice oil, which is present in globular bodies within
the juice sacs; and peel oil. Both juice oil and peel oil contribute oil-
soluble compounds (Cabral et al., 2010). The essential oil of orange juice
has been studied the most extensively, with approximately 220 volatile
compounds having been identified among its components. The oil con-
sists of up to 90% terpene hydrocarbons, with limonene being the most
abundant compound, and less than 2% aldehydes, alcohols, and esters
(Cabral et al., 2010). Flavor reconstitution experiments have reported
that the following combination of 22 volatiles, at appropriate concentra-
tions, reconstructed the aroma of orange very closely (Rouseff et al.,
2009): acetaldehyde, ethyl-2-methyl propanoate, (R)-R-pinene, ethyl
butanoate, (S)-ethyl-2-methyl butanoate, hexanal, (Z)-hex-3-enal,
­myrcene, (R)-limonene, 3-methyl butanol, 2-methyl butanol, ethyl hex-
anoate, octanal, oct-1-en-3-one, nonanal, decanal, (S)-linalool, butanoic
acid, (R)-ethyl-3-hydroxyhexanoate, (E,E)-deca-2,4-dienal, trans-4,5-
epoxy-(E)-dec-2-enal, and vanillin (Buettner and Schieberle, 2001).
Research on fresh ‘Mor’ mandarins has shown that the compounds
hexanal, ethyl-2-methyl butanoate, limonene, linalool, and β-myrcene

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provide green, fruity, citrus, floral, and musty aromas, respectively, all
of which c­ ontribute to mandarin flavor (Tietel et al., 2011).

16.2.6 Grape
Several volatile compounds have been identified in Vitis vinifera berries, with
these compounds mainly being present in the skin. These volatiles include
terpenes, norisoprenoids, aliphatic and aromatic alcohols and  ­aldehydes,
esters, and benzene derivatives (Ferrandino et al., 2012). Most of the infor-
mation available for this species has been obtained for wine varieties, leav-
ing a gap in the information available for table grape cultivars, especially
those consumed after a long-term storage period. Based on volatile com-
position, grape germplasm can be divided into three groups: Vitis labrusca
and its hybrids, which is characterized by abundant esters; Muscat cultivars
of V.  vinifera, with high concentrations of terpenoids; and neutral grapes,
mainly from cultivars of V. vinifera. The aroma of this latter group is mainly
composed of very low concentrations of C6 alcohols and aldehydes, benzene
derivatives, esters, and nonglycosylated monoterpenes and sesquiterpenes
(Yang et al., 2011).
Monoterpenes, in the form of free volatiles and glycoside conjugates
of monoterpene alcohols, contribute to the grape and wine aromas of sev-
eral cultivars. Typical monoterpenol components of aroma-rich grape vari-
eties are S-linalool, geraniol, nerol, citronellol, and α-terpineol (Defilippi
et al., 2009). C6 compounds impart grassy and herbaceous notes. Aliphatic
aldehydes, such as octanal, decanal, and (Z)-2-heptenal, impart a citrus-
like aroma, whereas furfural and benzaldehyde impart an almond aroma
(Ferrandino et al., 2012). Some volatile thiols, such as 4-mercapto-
4-­methylpentan-2-one, are considered to be impact odorants in Sauvignon
blanc wines, producing aromas that are reminiscent of box tree, passion
fruit, broom, and black current bud. Alternatively, 3-mercaptohexan-1-ol
and 3-mercaptohexyl acetate contribute to the aromas of passion fruit,
grapefruit, and citrus (Coetzee and du Toit, 2012).

16.2.7 Peach
More than 100 volatile compounds have been identified in peach, including
aldehydes, alcohols, alkanes, ketones, lactones, terpenes, and esters (Xi
et al., 2012). Among these compounds, lactones, in association with other
­volatiles, such as C6 aldehydes and alcohols and terpenes, play important
roles in peach aroma (Aubert et al., 2003; Cano-Salazar et al., 2013). Lactones,
particularly γ- and δ-decalactones, and esters, such as hexyl ­acetate and
(Z)-3-hexenyl acetate, are responsible for fruity aromas (Xi  et  al., 2012).

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Alternatively, C6 aldehydes and alcohols, such as n-hexanal, (E)-2-hexenal,

and (E)-2-hexenol, contribute green notes to the peach aroma (Zhang et al.,
2010; Xi et al., 2012).

16.2.8 Banana
More than 250 volatile compounds have been identified in banana culti-
vars (Imahori et al., 2013). A mixture of these compounds—mainly esters,
alcohols, and ketones—gives bananas their characteristic aroma (Brat
et al., 2004). Bananas mainly produce isoamyl and isobutyl esters, such as
3-methyl butyl and 2-methyl propyl esters of acetate and b ­ utanoate. These
esters are considered to be the major contributors to the aroma of bananas
(Jayanty et al., 2002), providing the “fruity” top notes (Wendakoon et al.,
2006). The alcohols pentan-2-ol, 3-methyl b ­ utanol, and 2-methyl p­ ropanol
also contribute to the succulent character of banana aroma, and the ketone
pentan-2-one, which is one of the major volatile compounds of bananas,
imparts the characteristic banana flavor (Brat et al., 2004).

16.3  Biosynthesis and Regulation of

Aroma Volatiles in Fruit

16.3.1 Biosynthetic Pathways of Aroma Volatiles

There are several pathways involved in the biosynthesis of the diverse
chemical classes of volatile compounds produced by fresh fruit. Although
some pathways of the synthesis of major volatiles are not fully understood,
it is widely assumed that aroma biosynthesis is related to the metabolism
of three main groups of precursors: fatty acids, amino acids, and carbohy-
drates. Fatty acids and amino acids serve as precursors for the biosynthesis
of the chemical groups, such as alcohols, aldehydes, and esters, which are
common in several fruits. Alternatively, carbohydrate metabolism may be
related indirectly to the formation of these groups and directly involved
in the biosynthesis of carbohydrate-derived aroma compounds, such as
­terpenoids and furanones.

16.3.1.1  Fatty Acid Metabolism

Saturated and unsaturated fatty acids are the major precursors of aroma
volatiles in fruits. They are metabolized through two major pathways,
β-oxidation and the lipoxygenase (LOX) pathway, to form straight-chain

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esters, alcohols, aldehydes, ketones, acids, and lactones. β-Oxidation is

considered the main metabolic pathway that produces aroma volatiles in
intact fruit, whereas the LOX pathway plays a major role in the production of
these compounds following tissue disruption. However, according to some
studies, the LOX pathway might function as an alternative to β-oxidation
during fruit ripening if allowed by a higher membrane ­permeability and an
increased availability of precursors (Sanz and Pérez, 2010).
In the β-oxidation pathway, successive removal of C2 units (acetyl
CoA) from saturated and unsaturated fatty acids leads, through several
mechanisms, to the formation of linear short- and medium-chain carboxylic
acids, aldehydes, and alcohols (Osorio et al., 2010). Moreover, acyl CoAs
generated through β-oxidation and alcohols are metabolized by alcohol
acyltransferases (AATs), producing a wide range of esters that contribute
to the aroma of nearly all fruits, as reviewed above (Schwab et al., 2008).
The biosynthesis of lactones or alkanolides is also associated with the
β-oxidation pathway, which is considered to act in concert with the LOX
pathway to form γ-(4) or δ-(5) lactones from their corresponding 4- or
5-hydroxy carboxylic acids (Sanz et al., 1997; Osorio et al., 2010).
In the LOX pathway, linoleic (18:2) and linolenic (18:3) acids serve as
precursors for the formation of C6 and C9 aldehydes and alcohols (Zhang
et al., 2010). Their biosynthesis mainly involves the action of four enzymes:
lipoxygenase (LOX), hydroperoxide lyase (HPL), 3Z,2E-enal isomerase,
and alcohol dehydrogenase (ADH). LOX enzymes, classified as 9-LOX
and 13-LOX to reflect their positional specificity with regard to fatty acid
oxygenation, catalyze the hydroperoxidation of linoleic and linolenic acids
to form 9- and 13-hydroperoxides (Bai et al., 2011). LOX p ­ roducts are fur-
ther metabolized, through HPL action, to C6 and C9 aldehydes and corre-
sponding ω -oxo acids (Sanz and Pérez, 2010). These C6 and C9 aldehydes
can be isomerized either by the action of 3Z,2E-enal isomerase or through
a nonenzymatic process (Schwab et al., 2008). Lastly, these aldehydes can
then be reduced by ADH to form the corresponding alcohols. Thus, LOX is
regarded as a key enzyme in the formation of straight-chain volatile com-
pounds from lipids, playing a major role in determining the composition of
fruit aroma. Research on LOX gene family members has been conducted
on several fruit species, such as apple (Schaffer et al., 2007), kiwifruit
(Zhang et al., 2009), and tomato (Chen et al., 2004). These studies have
provided evidence that LOX activity seems to be highly regulated at the
level of gene expression. For instance, the LOX gene family in tomato
includes at least five isoforms (TomloxA, TomloxB, TomloxC, TomloxD, and
TomloxE), which have diverse physiological functions. TomloxC, which is
detectable after the onset of ripening, is almost exclusively devoted to
the generation of C6 aldehydes and alcohols during tomato fruit ripening
(Chen et al., 2004).

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16.3.1.2  Amino Acid Metabolism

Amino acids serve as precursors for the biosynthesis of a large proportion of
aroma volatiles, including aliphatic, branched, and aromatic esters, alcohols,
carbonyls, and acids. Especially important are branched-chain volatile com-
pounds generated from the branched-chain amino acids leucine, ­isoleucine,
valine, alanine, and aspartic acid (Echeverría et al., 2004b; Schwab et al.,
2008). For instance, leucine is a precursor of some important v­ olatile com-
pounds such as 3-methyl butanal, 3-methyl butanol, and 3-methyl butanoic
acid, which have been found to influence the aroma of strawberry, tomato,
and grape varieties (Schwab et al., 2008); 2-­isobutylthiazole in tomato is
formed from valine (Osorio et al., 2010); and isoleucine has been reported
to be a precursor for 2-methyl butanol and 2-methyl butyric acid in melon,
apple, and strawberry (Sanz and Pérez, 2010).
The transformation of branched-chain amino acids into volatile com-
pounds is initiated with a transamination that is catalyzed by aminotrans-
ferases, forming 2-ketoacids. These are then metabolized into carboxylic
acids and aldehydes through decarboxylation or reduced to 2-hydroxyacids
(Schwab et al., 2008). The aldehydes can then be reduced by ADH to form
the corresponding alcohols. The aromatic amino acid phenyl alanine can
also be metabolized through this pathway, forming 2-phenyl acetyl-CoA,
which can be converted into a variety of alcohols and esters or reduced
to 2-phenyl ethanol and metabolized into 2-phenyl ethyl esters (Sanz and
Pérez, 2010). On the other hand, phenyl alanine is metabolized via a series
of complex pathways into phenyl propanoid and benzenoid compounds.
These nonvolatile metabolites are reduced at the C9 position to form v­ olatile
aldehydes and alcohols (Osorio et al., 2010). Phenyl propanoid–related com-
pounds, such as phenyl acetaldehyde and 2-phenyl ethanol, are abundant in
various fruits, including strawberry, tomato, and grape varieties (Aubert
et al., 2005). In addition, benzenoids, such as methyl salicylate, have been
found to influence tomato flavor (Tikunov et al., 2005).

16.3.1.3  Ester Biosynthesis

Volatile esters, which comprise one of the largest primary groups of volatile
compounds that have been identified to contribute to fruit aroma, are gen-
erated by the enzymatic routes described above. The metabolism of fatty
acids and amino acids forms aldehydes, which are reduced to alcohols in a
reversible reaction that is catalyzed by ADH (Díaz de León-Sánchez et al.,
2009). Alcohols are then esterified by AAT, which transfers an acyl moi-
ety from acetyl-CoA into the corresponding alcohol, forming an ester and
free CoA (Holland et al., 2005). Different combinations of alcohols and acyl-
CoAs result in a broad range of esters that account for the diversity of esters
produced by fruits (Shan et al., 2012). In general, fatty acid metabolism

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produces straight-chain esters, whereas amino acids are precursors of

branched-chain volatile esters (Altisent et al., 2009).
Research on ester-forming capacity through feeding studies in sev-
eral fruits has shown that the formation of volatile esters is dependent on the
availability of the corresponding substrates rather than on enzyme activity
(Dixon and Hewett, 2000; Dudareva et al., 2004; Song and Forney, 2008).
This is particularly true in bananas, where only the availability of substrates
from amino acid metabolism explains the composition of branched-chain
esters in banana aroma (Wyllie and Fellman, 2000).
The last two steps in the formation of volatile esters have been exten-
sively studied in terms of enzyme activities and expression of the corre-
sponding genes. Progress in the characterization of ADH and AAT enzymes
and genes responsible for ester biosynthesis has been reviewed by Defilippi
et al. (2009) and Song and Forney (2008). Some of the relevant characteris-
tics of these enzymes and their respective genes are now discussed.
The ADH gene family has been characterized in several fruit
­species, including apple (Reid et al., 1996), apricot (González-Agüero et al.,
2009), peach (Zhang et al., 2010), melon (Manríquez et al., 2006), tomato
(Longhurst et al., 1990), and grape (Tesniere and Verries, 2000). ADH genes
involved in the production of aroma are expressed in a developmentally reg-
ulated manner, specifically during fruit ripening (Schwab et al., 2008). Two
highly divergent ADH genes (CmADH1 and CmADH2) have been found to
be upregulated during melon ripening (Manriquez et al., 2006), and a strong
increase in VvADH2 transcript accumulation has been observed in grapes
at véraison (the onset of ripening) (Tesniere and Verries, 2000). In tomato,
LeADH2, one of the two ADH genes identified, accumulates during fruit
ripening (Longhurst et al., 1990), and its overexpression leads to increases
in the levels of some volatile alcohols, particularly Z-3-hexenol, improving
tomato flavor (Speirs et al., 1998).
The aroma-related AAT enzyme has been a major focus of research
in the past. AATs directly involved in volatile generation have been found in
many fruit species, including apple (Fellman et al., 2000; Echeverría et al.,
2004b; Defilippi et al., 2005b; Holland et al., 2005), melon (Shalit et al., 2001;
El-Sharkawy et al., 2005), strawberry (Aharoni et al., 2000; Beekwilder
et al., 2004), banana (Harada et al., 1985; Beekwilder et al., 2004), and grape
(Wang and DeLuca, 2005). The results of a number of precursor feeding
experiments have demonstrated that different substrate specificities of
AATs may account for the observed variation in volatile ester composition
in several fruits. For instance, the AAT family is composed of at least four
members in melon. Among these, Cm-AAT1 protein produces a wide range
of short- and long-chain acyl esters but is strongly predisposed to form E-2-
hexenyl acetate and hexyl hexanoate. In contrast, Cm-AAT3 shows a very
strong preference for producing benzyl acetate, and Cm-AAT4 is almost
entirely responsible for the formation of acetates, particularly cinnamoyl

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acetate (El-Sharkawy et al., 2005). Similarly, MdAAT2 ­ preferentially

produces pentyl acetate and hexyl acetate in apple (Li et al., 2006), and
strawberry AAT preferentially accepts geraniol, 1-heptanol, 1-octanol, and
1-nonanol to produce its corresponding esters (Song and Forney, 2008).

16.3.1.4  Carbohydrate Metabolism

Furanones and terpenes are two main groups of aroma compounds that
originate directly from carbohydrates. Both chemical groups contribute to
the flavor of many fruits and often determine their characteristic aroma.
Furanones, which have a pleasant sweet aroma, have been inten-
sively studied due to their attractive sensory properties. Among these com-
pounds, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (furaneol) is defined as
one of the key odorants in pineapples (Tokitomo et al., 2005) and is also
considered, together with its derivative, 2,5-dimethyl-4-methoxy-3(2H)-
furanone (mesifurane), to be a character-impact compound in the aroma
and flavor of fresh strawberry (Forney et al., 2000). The biosynthetic path-
way of furaneol has not been fully elucidated; however, substantial progress
has been made in recent years. Experiments with incorporation of labeled
precursors have revealed that D-fructose-1,6-diphosphate is the natural
precursor of furaneol biosynthesis and its derivatives (Roscher et al., 1998).
Studies in strawberry fruit have shown that phosphorylated carbohydrate
is converted by phosphate and water elimination to 4-hydroxy-5-methyl-
2-methylene-3(2H)-furanone, which is reduced to furaneol by an enone
oxidoreductase enzyme (FaEO) (Schwab, 2013). Mesifurane is formed
through the methylation of furaneol catalyzed by an O-methyltransferase
(FaOMT), which might use S-methyl-adenosyl-L-methionine (SAM) as a
donor of the methyl group in the 4-methoxy compound of mesifurane (Wein
et al., 2002).
In the case of terpenoids, they comprise one of the most diverse fami-
lies of natural products, many of which are nonvolatiles and contribute to
important plant processes (Schwab et al., 2008). The volatile terpenoids,
monoterpenes (C10) and sesquiterpenes (C15), play an important role in
the overall flavor of several fruits; however, monoterpenes are considered
the main fruit aroma components in the terpenoid family. For instance,
monoterpenes that have been found to greatly influence fruit aroma are
R-limonene in citrus fruit (Cabral et al., 2010), geraniol and citronellol in
Muscat grapes (Luan et al., 2005), and S-linalool in tomato fruit (Lewinsohn
et al., 2001). Terpenoids are produced through two biosynthetic routes, the
mevalonate (MVA) and methyl erythritol phosphate (MEP) pathways, both
of which lead to the formation of common intermediates for all terpenoids,
isopentenyl diphosphate (IPP), and its isomer, dimethyl allyl diphosphate
(DMAPP) (Sanz and Pérez, 2010). The MVA pathway, which is active

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in cytosol and starts from acetyl-CoA, is generally considered to supply

­precursors for the biosynthesis of sesquiterpenes. On the other hand, the
MEP pathway, which is active in the plastids and starts from pyruvate and
glyceraldehyde-3-phosphate, might supply precursors for the production of
monoterpenes (Dudareva et al., 2004). These two pathways appear to work
independently, although there is increasing evidence that limited unidirec-
tional exchange of intermediates occurs between cellular compartments
(Schuhr et al., 2003). In the next phase, IPP and DMAPP are metabolized
by prenyl transferases into geranyl diphosphate (C10), farnesyl diphos-
phate (C15), and geranyl geranyl diphosphate (C20), which are precursors of
monoterpenes, sesquiterpenes, and diterpenes, respectively. Lastly, these
prenyl diphosphates are transformed to the final terpenoids by terpene syn-
thases (Dudareva et al., 2004; Pichersky et al., 2006; Schwab et al., 2008).
Although many volatile terpenes are synthesized through this pathway,
others are formed through the transformation of initial volatile compounds
by oxidation, dehydrogenation, acylation, and other reaction types. Such
reactions result in compounds with enhanced volatility or different olfac-
tory ­properties (Dudareva et al., 2004).
As primary metabolites, carotenoids also form a group of volatile
­terpenes, apocarotenoids, or norisoprenoids. These terpenoids are ­present
at relatively low concentrations but have a major impact on fruit aroma.
Apocarotenoids with 13 carbon atoms are presumably generated in plastids
by carotenoid cleavage dioxygenases (CCDs), which catalyze the oxida-
tive cleavage of carotenoids (tetraterpenes, C 40) at specific locations on the
backbone (Gapper et al., 2013). Studies in tomato and watermelon have pro-
vided evidence that carotenoids not only are key pigments in these fruits,
but also serve as precursors for important aroma volatiles, as lycopene is
presumably for geranial in these fruits (Lewinsohn et al., 2005).

16.3.2  Aroma Modulation during Fruit Ripening

Ripening of fleshy fruits involves a series of molecular, biochemical, and
physiological processes that lead to an overall modification of the fruit,
including changes in color and texture and metabolic changes related to
flavor and nutrient composition, generally associated with accumulation of
sugars and aroma compounds (Klee and Giovannoni, 2011). The synthe-
sis of aroma volatiles is therefore associated with fruit ripening, a stage in
which the concentrations of volatile compounds change both qualitatively
and quantitatively, depending on the species or varieties. In general, during
ripening, the aroma profile of fruits changes from an abundance of grassy-
note compounds, such as aldehydes and alcohols, to a profile dominated
by fruity-note compounds, given by esters in apples (Fellman et al., 2000),

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lactones in peaches (Zhang et al., 2010), and both of these compounds, in

addition to furanones, in strawberries (Ménager et al., 2004).
Depending on their respiratory behavior, fruits can be divided into two
groups, climacteric and nonclimacteric. In climacteric fruit, a significant
increase in respiration is linked to a rise of ethylene production at the onset
of ripening, where this hormone is necessary for triggering physiological
changes in fruit attributes. In contrast, a sharp increase in respiration and
ethylene production does not occur in nonclimacteric fruit, such as grapes
and citrus fruit, and therefore ripening-related changes involve mechanisms
that are independent of ethylene (Defilippi et al., 2009; Gapper et al., 2013).
For climacteric fruit, ethylene is considered to be a critical factor in determin-
ing fruit aroma. This relationship is based on evidence that climacteric fruits,
such as apple, melon, and banana, exhibit maximum volatile compound pro-
duction near their climacteric peaks (Dixon and Hewett, 2000; Fellman et al.,
2000; El-Sharkawy et al., 2005). In apples, this peak occurs together with an
increase in ester production, an effect that arises mainly from a greater avail-
ability of substrates, as confirmed by several studies (Bartley, 1985; Song and
Bangerth, 2003), in which enhanced accumulation of fatty acids (FAs) has
been observed during the climacteric period, concomitant with an increase
in aroma production. Although early studies postulated that these fatty acids
could derive from the catabolism of membranes or other lipid “storage” pools
(Paillard, 1986; Fellman et al., 2000), with the work of Song and Bangerth
(2003), the hypothesis that a high respiration via ATP and de novo FA biosyn-
thesis related to the presence of autocatalytic ethylene production occurring
at the onset of the climacteric period leads to high volatile aroma production
appears to be more valid based on their results.
The effect of ethylene on aroma production has been studied exten-
sively through the use of ethylene inhibitors such as aminoethoxyvi-
nylglycine (AVG) and 1-methylcyclopropene (1-MCP), which inhibit the
biosynthesis and action of this hormone, respectively. Both technologies
have been shown to have a detrimental effect on the production of ripening-
related volatile compounds, as evidenced by the decreased ester production
observed in several fruits to which these inhibitors have been applied, such
as banana (Golding et al., 1998), plum (Abdi et al., 1998), and apple (Defilippi
et al., 2004; Mattheis et al., 2005; Ferenczi et al., 2006). These results
indicate that ester biosynthesis requires continuous ethylene ­activity. As
expected, many of the genes that encode enzymes involved in volatile pro-
duction are under the control of ethylene at the transcriptional level. Fatty
acid metabolism seems to possess important transcriptional points that are
highly regulated by ethylene, as observed in apple, in which the last step of
volatile ester production is extremely induced by ethylene via its impact on
AAT transcript accumulation (Defilippi et al., 2005a; Schaffer et al., 2007).
In peach, ethylene also induces the accumulation of linoleic and linolenic
acids by its influence on the upregulation of fatty acid desaturase genes

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(PpFAD1 and PpFAD2), in addition to its contribution to the ­production of

grassy aroma aldehydes and alcohols, with this latter effect inducing the
downregulation of certain LOX, HPL, and ADH genes in this fruit (Zhang
et al., 2010).
The association between ethylene and aroma production has also
been studied through the use of transgenic fruits with suppressed ethyl-
ene biosynthesis. Transgenic lines with the ACS (ACC synthase; ACC–1-
aminocyclopropane-1-carboxylic acid) or ACO (ACC oxidase) enzymes
blocked by the antisense technique to reduce autocatalytic ethylene pro-
duction have been developed in apple (Dandekar et al., 2004; Schaffer et al.,
2007), melon (Bauchot et al., 1998), and tomato (Oeller et al., 1991), result-
ing, in all these studies, in fruit with lower aroma levels. The synthesis of
volatile esters was found to be reduced by 65%–70% in transgenic apples,
although their precursors, aldehydes and alcohols, were not ­significantly
suppressed (Dandekar et al., 2004). These results suggest that downstream
steps in ester biosynthetic pathways are under the control of ethylene,
which was confirmed later, as described above, with the work of Defilippi
et al. (2005a) and Schaffer et al. (2007), who found that the enzyme respon-
sible for ester production, AAT, is strongly regulated by this hormone.
Similar levels of ester inhibition were previously observed in climacteric
ACO antisense transgenic melon (Bauchot et al., 1998). Interestingly,
­f urther studies using disks of ACO antisense melon in the presence of vari-
ous precursors and treated with 1-MCP showed that the reduction of fatty
acids and aldehydes might be ethylene dependent, whereas the last step of
ester production, only partly inhibited, has both ethylene-independent and
ethylene-dependent components (Flores et al., 2002).

16.4  Influence of Pre- and Postharvest

Factors on Fruit Aroma

16.4.1  Preharvest Factors

Because the aroma profile in fruit is determined by intrinsic and exogenous
factors, and due to the complex interactions among them, describing their
influences on this quality trait is a challenging task that we approach in this
section by summarizing the most relevant factors affecting overall aroma
formation in fresh fruit.

16.4.1.1 Genotype
Fruit aroma is largely determined by the fruit’s genetic background, and
accordingly, significant differences in the composition and concentra-
tions of volatile compounds have been observed among cultivars within

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a fruit species. In apple, for example, early studies classified varieties

based on the type and amount of volatile compounds that they mainly pro-
duce (Dixon and Hewett, 2000), while later studies have been conducted
to elucidate the mechanisms responsible for the differences observed
in the accumulation of volatile compounds among apple cultivars and
strains. In this way, the characterization of ester production by one of
the most aromatic apples, ‘Fuji’, and one of the least aromatic, ‘Granny
Smith’, performed through comparative volatile determination, feeding
experiments, and enzyme–substrate specificity, has shown that distinct
­activity levels and substrate specificity of A AT enzymes may account for
the differences in volatile ester production between the cultivars (Holland
et al., 2005). The A AT enzyme, which is present in the peel and flesh of
‘Fuji’ apples, accepts a wide range of alcohols as substrates, whereas A AT
detected only in the peel of ‘Granny Smith’ uses almost exclusively hexa-
nol and cis-3-hexenol to produce the corresponding esters (Holland et al.,
2005). An interesting relationship between peel coloration and ester pro-
duction has been found in different strains of ‘Delicious’ and ‘Fuji’ apples
(Fellman et al., 2000; Iglesias et al., 2012). For both varieties, the most
colored (blushed) strains produced fewer volatile esters than the least col-
ored (striped) strains. This difference is mainly attributed to the limited
substrate availability in blushed apples, resulting from the sequestration
of acyl moieties in the anthocyanin molecules found in peel cell vacuoles
(Fellman et al., 2000).
In strawberries, considerable differences exist between the volatile
compound profiles of wild (Fragaria vesca L.) and cultivated (Fragaria ×
ananassa Duch.) plants. The cultivated octaploid species (2n = 56) has a
higher yield but a less characteristic strawberry aroma than the wild dip-
loid (2n = 14) species (Zabetakis and Holden, 1997). The main difference
in aroma between species relies on the ester and terpenoid profiles. While
acetate esters, such as methyl butanoate, ethyl butanoate, ethyl hexanoate,
and methyl-2-methyl butanoate, and the monoterpene S-linalool and the
sesquiterpene nerolidol all contribute to the fruity aroma of cultivated vari-
eties, the aroma of wild varieties is dominated mainly by the ester methyl
anthranilate, which imparts a marked floral odor in addition to a fruity note,
and by olefinic monoterpenes and myrtenyl acetate, which contribute the
turpentine-like, woody, resinous odor of this species (Aharoni et al., 2004;
Dong et al., 2013).
Many other studies have been performed to determine the aroma
­profiles of different cultivars in other fruits, such as tomato (Langlois et al.,
1996), peach (Wang et al., 2009), and melon (Aubert and Bourger, 2004).
Interestingly, phenotypic differences in volatile production of melons are
apparently correlated to their storage life. Cantalupensis melons tend to be
more flavored and have a shorter storage life than those from the Reticulatus
group (Song and Forney, 2008). Similarly, among the Cantalupensis group,

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Charentais cantaloupe melon, bred for longer shelf life, has much lower
concentrations of volatile compounds than short-shelf-life cultivars (Aubert
and Bourger, 2004).

16.4.1.2  Growing Conditions

Due to their effects on overall plant metabolism, environmental conditions
and cultural practices greatly influence the production of aroma compounds
in fruits. Among climatic conditions, temperature, light (quality and inten-
sity), and rainfall are factors that characterize a specific growing location,
contributing to the characteristic aroma of a fruit genotype. Although it
is widely known that the growing area is a multivariable parameter influ-
encing fruit quality, its effect on fruit aroma has been poorly documented.
Recently, Ferrandino et al. (2012) found that the volatile profile of V. ­vinifera
cv. ‘Nebbiolo’ was greatly influenced by the area, or “terroir,” where it was
grown. Even in a relatively homogenous geographic area, significant dif-
ferences among growing locations were observed in the accumulation of
esters in ‘Nebbiolo’ berries. Also in grape, the effect of light or sun exposure
by itself on volatile production has been studied by comparing natural light
conditions with artificial shading treatments. Thus, shaded clusters (20%
sun exposure) of two Chilean Muscat grape cultivars were found to have
significantly lower contents of terpenols, particularly linalool. In contrast,
partially shaded vines (50% sun exposure) accumulated the highest levels
of terpenols, resulting in grapes with a better Muscat aroma (Belancic
et  al., 1997). Similar trials performed with ‘Muscat of Frontignan’ and
‘Syrah’ grapes have shown that in addition to the reduction in monoterpe-
nol contents, C13 norisoprenoid accumulation also decreased in artificially
shaded grapes (Bureau et al., 2000a, 2000b). The effect of sun exposure on
C13 norisoprenoid content has been associated with the light-induced bio-
synthesis of carotenoids, which are direct precursors of these compounds
(Schultz et al., 1998).
In the ‘Delicious’ apple variety, acetate ester production has also
been found to be influenced by shading, with lower acetate ester contents
detected in fruit with greater than or less than 53% sun exposure (Miller
et al., 1998). In contrast to grapes and apples, for which partial shading
is beneficial, strawberries do not benefit from shading. Shading of 47%
has been found to result in a significant decrease in the accumulation of
­hexenal, hexanal, ethyl methyl butyrate, and methyl butyrate in strawber-
ries, compared to unshaded fruits (Watson et al., 2002). These findings
suggest that the effect of sun exposure on fruit aroma composition depends
on the species, and complex mechanisms might be involved. However, it
should be noted that all pathways involved in aroma biosynthesis start
from primary metabolic products generated by photosynthesis, and there-
fore, under optimum conditions, sunlight might positively induce aroma

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production, while an excess of radiation and heat could cause stress, either
by ­dehydration or by temperature increase in the plant, negatively affecting
aroma biosynthesis.
Cultural practices can also greatly influence fruit aroma, often reduc-
ing the quality of this attribute due to the use of crop management sys-
tems based almost exclusively on crop yield. Mattheis and Fellman (1999)
reviewed several studies reporting detrimental effects of high crop load and
the use of technologies for the control of diseases and pests on apple aroma.
As water status is a key factor in plant physiology, modulating pri-
mary and secondary metabolism, it is expected that irrigation might induce
changes in the biosynthesis of aroma volatiles. Veit-Köhler et al. (1999)
found that ‘Vanessa’ tomatoes produced a higher accumulation of C6 alde-
hydes, such as hexanal, (Z)-3-hexenal, and (E)-2-hexenal, under conditions
of lower water supply (without visible symptoms of water stress), resulting in
fruit with better quality without significant effects on fruit growth and yield.
The aroma potential and total content of C13 norisoprenoids were found to
increase in ‘Cabernet Sauvignon’ grapes grown with water deficit (Bindon
et al., 2007; Koundouras et al., 2009). An indirect effect of irrigation, particu-
larly on grape aroma, is its influence on shoot growth and leaf area, which
affect sunlight penetration and consequently aroma metabolism.
A practice that is often used to control soilborne diseases is g­ rafting.
It is known that yield and quality attributes of fruit can be affected by
grafting and the type of rootstock. For instance, grafting muskmelon onto
pumpkin has been found to reduce aromatic ester accumulation in this
fruit compared with melon grown on its own roots, as a result of decreased
activities of alcohol dehydrogenase and alcohol acyltransferase enzymes
(Chuan-qiang et al., 2011; Condurso et al., 2012).
Aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibi-
tor, is commonly used to avoid premature fruit drop and extend the har-
vest period, mainly for apples, to which this inhibitor is applied 1 month
before harvest (Escalada and Archbold, 2009). A significant reduction in
total volatile compound production after storage has been reported for
several cultivars of AVG-treated apples, whose decline in aroma produc-
tion was enhanced as storage time proceeded, and especially in those
stored under controlled atmosphere conditions (Bangerth and Streif,
1987; Mir et al., 1999).
Fertilization is also considered to affect fruit aroma. Studies in which
the influences of nutrients at various concentrations have been examined
have shown that suitable levels of potassium and nitrogen induced ester
production in muskmelon and strawberry, respectively, improving their
aroma (Lin et al., 2004; Ojeda-Real et al., 2009). In contrast, either a defi-
cient or excessive use of fertilizers had negative effects on aroma produc-
tion in these fruits. However, extreme levels of nutrients might be beneficial
for some species. For instance, the contribution of the C13 norisoprenoid

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1,1,6-trimethyl-1,2-dihydronaphthalene (TDN) is especially ­important for

the kerosene-like note in ‘Riesling’ wines, and its accumulation is affected
by fertilization with nitrogen. Thus, higher TDN concentrations in
‘Riesling’ wines resulted from a long-term nitrogen deficiency in these
vines (Linsenmeier and Lohnertz, 2007). Indirectly, for any fruit species,
whatever nutrient system is used will affect plant vigor and therefore matu-
rity at harvest, which greatly influences aroma production, as discussed in
Section 16.4.1.3.

16.4.1.3  Fruit Maturity

Maturity at harvest plays a major role in overall flavor, including fruit aroma
development, by greatly influencing flavor production during postharvest
ripening. Although harvesting fruit at optimal maturity might ensure
the maximum development of aroma volatiles for flavor, it usually results
in fruit with poor storability with practically no shelf life. To avoid these
undesirable outcomes, the anticipation of harvest time for most types of
fruit is a common commercial practice for increasing fruit storability and
marketability and minimizing damage produced by postharvest handling.
Thus, early harvested fruit develops a better texture, sugar/acid ratio, and
background color during storage, at the cost of reduced capacity to produce
aroma volatiles (Bangerth et al., 2012).
It has been observed for several fruit species that there is a time lag
until the early harvested fruit starts to produce ripening-related aroma
compounds; however, its full aroma spectrum will never attain that of fruit
harvested at a more mature stage (Mattheis and Fellman, 1999). In general,
the closer the harvest time is to the time of optimal fruit maturity, the less
time it takes for the fruit to recover its capability to produce aroma volatiles.
For instance, ‘Golden Delicious’ apples picked approximately 4 weeks prior
to their optimal harvest time have been shown to exhibit a small increase
in aroma production after 5 weeks of shelf life, whereas those harvested
approximately 2–3 weeks prior to their optimal maturity took 2 weeks of
shelf life to recover their aroma biosynthesis rates and even reached levels
similar to those of apples harvested at their optimal maturity (Song and
Bangerth, 1996).
Maturity at harvest also greatly influences the regeneration of aroma
production by fruits during storage, especially under long-term controlled
atmosphere (CA) and ultralow oxygen (ULO) conditions (Song and Forney,
2008). Previous studies in ‘Delicious’ apples have shown that the optimal
harvest time for maintaining aroma-generating capacity during CA is when
fruit are stored just prior to the beginning of the climacteric period. Similar
to the effect of maturity at harvest on postharvest ripening, the earlier the
harvest is before the climacteric rise in respiration, the more time that might
be required for the fruit to regenerate its ability to produce aroma volatiles

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during poststorage (Brackmann et al., 1993; Fellman et al., 2003). Thus,

the impact of fruit maturity at harvest on the fruit’s capacity to s­ ynthesize
volatiles is closely related to changes in the respiratory rate and ethylene
production. Song and Bangerth (1994, 1996) have related the poor ability
of early harvested apples to produce aroma volatiles to low respiration and
fatty acid levels. It seems that a considerable insensitivity of immature fruit
to ethylene results in a low respiration rate and, consequently, in low levels
of precursors, mainly ATP, required for fatty acid biosynthesis (Song and
Bangerth, 1996, 2003).

16.4.2  Postharvest Technologies

Many fruit commodities are subjected to several postharvest procedures
to maintain fruit quality and allow the fruit to be marketed for an extended
period of time. However, postharvest handling approaches to quality are
often based on maintaining or improving quality attributes such as texture,
color, and sugar/acid ratio, but not aroma. As most postharvest technologies
rely on diminishing metabolic rates of fruits, these procedures certainly
have a huge impact on the production of aroma volatiles, whose influence is
closely related to the previously discussed preharvest factors. Therefore, in
this section, we provide an overview of the impact of the postharvest tech-
nologies most widely used by the fruit industry on fruit aroma, highlighting
the main mechanisms involved in the complex process of aroma develop-
ment during the postharvest life of fruits.

16.4.2.1  Storage Temperature

Low-temperature storage is the primary and most common tool for delay-
ing ripening after harvest by reducing the respiration rate of fruit. Although
cold storage is effective in reducing postharvest fruit losses, this method
alters the production of aroma volatiles. In general, the lower the temper-
ature is during storage, the lower the production of volatiles is by fruits
(Dixon and Hewett, 2000). However, the influence of temperature on the
production of aroma-related volatiles depends on the storage potential of
fruit species and cultivars. For instance, peach fruit is particularly prone to
develop chilling injury during cold storage, which is associated with several
physiological disorders, including flesh browning, mealiness, and usually a
loss of the characteristic flavor, among other effects (Xi et al., 2012). Zhang
et al. (2011) reported that chilling-injured peach (Prunus persica L. Batsch.
cv. ‘Hujingmilu’) had a significantly reduced capacity to produce fruity note
volatile compounds such as esters and lactones after storage at 5°C for up to
21 days, followed by storage at 20°C for 3 days. To overcome chilling injury,
several postharvest treatments have been proven to be effective in reducing

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chilling-related disorders during cold storage of stone fruits. Among these

treatments, a conditioning period at 20°C for 24–48 h after harvest and
before cold storage has been shown to be particularly effective, although
fruit softening occurs (Crisosto et al., 2004). This practice not only prevents
chilling injury, but also improves volatile compounds emissions. Thus, for
‘Early Rich’ peaches, 36 h of prestorage at 20°C prior to storage at –0.5°C for
20 days induced an increase in esters such as propyl acetate and pentyl ace-
tate, resulting in the conditioned fruit being perceived as sweeter than the
nonconditioned peach (Cano-Salazar et al., 2013). Although these authors
did not elucidate the mechanisms enhancing aroma production in prestored
fruit, the results of research performed by Xi et al. (2012) on peach (Prunus
persica L. Batsch cv. ‘Jinxiu’) subjected to intermittent warming by expos-
ing the fruit to 20°C for 1 day during every week of storage at 5°C suggest
that higher levels of fatty acids and an increased ­activity of AAT might be
responsible for enhanced aroma volatile production in ­conditioned peach.
The adverse effect of cold storage on fruit aroma has also been
documented in tomato and mandarin. Díaz de León-Sánchez et al. (2009)
reported that higher ratios of 3-methyl butanal/3-methyl butanol and
hexanal/­hexanol, as well as an increase in trans-3-hexenol levels, were the
most likely causes of the off-flavor of cold-stored tomatoes; notably, all of
these observed effects were associated with a reduced ADH enzyme activ-
ity. Similarly, a reduction in ADH and HPL activities has been associated
with an inhibition of C6 aldehyde and alcohol aroma volatiles in chill-injured
tomatoes (Bai et al., 2011). In mandarins, an interesting approach has been
used by Obenland et al. (2013) to identify the cause of the diminished
mandarin-like flavor induced by storage temperature. In contrast to the
approaches taken in other studies to simulate storage temperatures follow-
ing marketing of mandarins, these authors determined independently the
effects of cold and warm temperatures on mandarin aroma. Interestingly,
they found that holding temperature at 20°C after cold storage at 5°C was the
cause of the flavor quality loss in mandarins, mainly because of enhanced
production of volatile alcohols (3-methyl-1-butanol and 2-methyl-1-butanol)
and ethyl esters, which contribute to an overripe and pronounced off-flavor
in this fruit. The authors suggested that controlling the temperature after
packing (5°C–10°C) could be an effective tool in preventing flavor quality
loss in many mandarin varieties.
Dixon and Hewett (2000) reviewed several studies that document the
influence of storage temperature on the production of aroma volatiles in
apples. All of those studies reported decreases in volatile contents, particu-
larly esters, in cold-stored apples, and not surprisingly, the influence of cold
storage was reported to be highly dependent on the fruit variety and the
length of exposure. In general, lower production of aroma-related volatiles
by apples was observed for longer cold storage periods. This reduced ability
to produce volatile esters was found to be associated with decreased AAT

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activity in ‘Rome’ apples (Fellman et al., 1993) and presumably to reduced

availability of substrates in cold-stored fruits.

16.4.2.2  Storage Atmosphere

The atmosphere composition during storage, particularly O2 and CO2
­c oncentrations, greatly influences the aroma of fresh fruits. Compared
to conventional regular air conditions, controlled atmosphere (CA)
storage has been increasingly adopted by the fruit industry for better
maintenance of fruit attributes, such as firmness, color, and acidity, and
for reducing the incidence of physiological disorders during long-term
storage. However, the use of this technology, mainly at ultralow oxy-
gen (ULO) concentrations, has a negative impact on the aroma quality
of fruits. Studies on the effect of CA storage on aroma production have
focused primarily on apples and have shown that prolonged low O2 stor-
age delays the onset of biosynthesis of aroma compounds and greatly
reduces their production, particularly for esters, during subsequent rip-
ening (Streif and Bangerth, 1988; Brackmann et al., 1993; Fellman et al.,
2000; Mattheis et al., 2005). This suppressed capacity to produce aroma
volatiles is considered to be the main cause of the off-flavor of CA-stored
apples. Another major finding of these studies is that the severity of the
effect of CA on fruit aroma is largely dependent on the composition of the
storage atmosphere and the length of storage. In general, for CA-stored
apples, a higher suppression of aroma results from lower O2 and higher
CO2 levels, as well as longer storage durations (Fellman et al., 2000).
Thus, for ‘Golden Delicious’ apples, all CA conditions tested (3% CO2 +
3% O2 , 3% CO2 + 1% O2 , and <1% CO2 + 1% O2) significantly suppressed
aroma volatile production compared with regular air conditions, and the
greatest detrimental effect on apple aroma was found for ULO (1% O2
+ 3% CO2) storage, the suppressive effect of which increased with the
length of the storage period (Brackmann et al., 1993). These authors
also demonstrated that low O2 conditions reduced the concentration of
total fatty acids in apples, indicating that a lack of these precursors is
most likely the main reason why apples stored under CA conditions have
reduced volatile production. However, similar to the effect observed in
fruit harvested before reaching optimal maturity, low fatty acid avail-
ability results from low respiration rates, which is linked to disturbed
ethylene sensitivity in fruits, suggesting that the inhibition of ethylene
action and synthesis by CA storage might be the main mechanism by
which aroma biosynthesis is reduced in CA-stored apples and other
fruits. Nevertheless, unlike preclimacteric apples, in which e­ xogenous
ethylene treatment is an option to induce production of aroma volatiles,
fruit fails to respond to exogenous ethylene treatment after long-term CA

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 F RES H F RUIT AROMA

storage (Song and Bangerth, 2003). Therefore, alternative techniques

that stimulate respiration and ethylene production might be necessary
to enhance formation of volatiles after CA storage. For instance, an extra
period (4 weeks) of cold storage under regular air conditions has been
shown to induce a partial recovery of aroma production in ‘Fuji’ apples
after 30 weeks of storage at 1°C under ULO conditions (1% O2 + 1% CO2).
This recovery has been related to the high activity of the LOX enzyme
during the conditioning period (Altisent et al., 2009).
In addition to the influence of CA composition on total aroma pro-
duction, several studies have reported that O2 and CO2 levels also greatly
influence the emission of individual volatile compounds (Brackmann et al.,
1993; Echeverría et al., 2004a; Lara et al., 2006). These studies have shown
that the production of straight-chain esters by apples is highly suppressed
by low O2 conditions, which has been associated with an inhibition of the
activity of LOX, an O2 -requiring enzyme, under hypoxic conditions. In con-
trast, biosynthesis of branched-chain esters is reduced only at high CO2
concentrations, whose precursors derive mainly from amino acids gener-
ated by the tricarboxylic acid cycle, which is known to be inhibited by high
CO2 levels (Fellman et al., 2000). This effect on the aroma ester profile has
been observed even for moderate differences in O2 and CO2 concentrations.
Raffo et al. (2009) found significant differences between the effects of ULO
and dynamic controlled atmosphere (DCA) technologies on straight- and
branched-chain ester production in ‘Pinova’ apples stored up to 7 months at
1.3°C. While DCA (0.4%–0.6% O2 and 0.6%–0.8% CO2) storage, with lower O2
levels, greatly inhibited straight-chain ester biosynthesis, branched-chain
ester production was more strongly inhibited by ULO (1.5% O2 + 1.3% CO2)
conditions, with higher CO2 levels.
Modified atmosphere packaging (MAP) has been developed as an
alternative method to CA storage to prolong fruit shelf life. MAP containing
high levels of CO2 (10%–20%) is widely used to prevent decay and preserve
quality attributes of strawberries under cold storage (Pelayo et al., 2003).
However, under these conditions, especially if temperature control is lack-
ing, the development of off-flavors often occurs due to an accumulation of
fermentative metabolites, primarily acetaldehyde, ethanol, and ethyl esters
(Ke et al., 1994; Almenar et al., 2009). Among ethyl esters, the formation of
ethyl acetate in particular is induced by the enhanced production of ethanol
in enriched-CO2 atmospheres. Moreover, due to its low odor threshold and
a chemical-like aroma at high concentrations, ethyl acetate has been con-
sidered to contribute greatly to the off-flavor of strawberries under stress-
ful CO2 levels (Ke et al., 1994; Larsen and Watkins, 1995; Forney et al.,
2000). According to Pelayo-Zaldívar et al. (2007), a change in the synthesis
of methyl to ethyl esters induced by enriched-CO2 atmospheres might also
contribute to the loss of strawberry flavor during storage.

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16.4.2.3  Ethylene Control

We previously discussed the importance of ethylene in modulating changes
in the volatile profile of fruits. Several tools are available for reducing
­ethylene levels during storage, the most common being controlled atmo-
sphere storage, potassium permanganate scrubbers, and the application of
the ­ethylene inhibitor 1-methylcyclopropene (1-MCP).
The ripening and senescence processes regulated by ethylene can be
significantly delayed through the application of 1-MCP. This cyclic ­olefinic
compound binds to ethylene receptors with much more affinity than
­ethylene itself, antagonizing its action. Postharvest use of 1-MCP, along
with cold storage, is a common commercial tool to prolong the shelf life
of several climacteric fruits. Due to its importance to the fruit industry, a
large number of studies have been published on the effectiveness of 1-MCP
with respect to the maintenance of quality attributes of fruits, such as flesh
firmness, skin color, sugar/acid ratio, and its impact on fruit aroma. Most
climacteric fruits have been found to exhibit reduced or delayed production
of several volatile compounds, particularly esters, after exposure to 1-MCP
(Golding et al., 1998; Fan and Mattheis, 1999; Kondo et al., 2005; Mattheis
et al., 2005; Rizzolo et al., 2005). The extent of this aroma inhibition varies
greatly among species and cultivars and depends on the concentration of
the inhibitor applied, as well as the length of storage (Raffo et al., 2009). As
mentioned earlier for other ethylene inhibition technologies, such as CA, a
recovery of volatile biosynthesis also takes place during ripening of 1-MCP-
treated fruits following removal from cold storage; however, the volatile
biosynthesis does not reach the levels observed in nontreated fruit. Thus,
for ‘Jonagold’ and ‘Red Chief Delicious’ apples, a partial volatile production
recovery was reached for up to 3 weeks holding at 22°C, after 2 or 3 months
in storage at 0°C in air, and after 7 months under CA ­conditions (Ferenczi
et al., 2006). As CA storage alone has been shown to have a strong effect in
reducing the production of volatiles, the effect of 1-MCP in inhibiting vola-
tile biosynthesis is expected to be enhanced in combination with CA stor-
age, and of course it is, as has been observed for ‘Gala’ and ‘Empire’ apples
(DeEll et al., 2005; Mattheis et al., 2005), in which CA storage further
suppressed aroma biosynthesis in treated fruit and prolonged the effect
of 1-MCP. On the other hand, studies analyzing the independent effects of
these two technologies on changes in aroma patterns in some species have
shown that 1-MCP induces far less aroma suppression than CA storage. For
instance, 1-MCP treatment reduced the content of hexyl acetate, a major
­volatile ester with a fruity, sweet aroma, by approximately 50% in ‘Tardibelle’
peaches, compared to the level in untreated peaches, in contrast to CA stor-
age, which reduced levels of this ester to values well below its detection
limit (Ortiz et al., 2010b). Similarly, 1-MCP-treated ‘Packham’s Triumph’
pears developed a better aroma after cold storage than fruits stored under

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CA conditions, being highly scored by panelists in terms of their capacity to

maintain this and other quality attributes (Moya-León et al., 2006).
Studies on the effect of 1-MCP on fruit aroma have also shown that
treatments with this compound influence not only total volatile production,
but also volatile profile. In ‘Pinova’ and ‘Gala’ apples (Mattheis et al., 2005;
Raffo et al., 2009), ‘Packham’s Triumph’ pears (Moya-León et al., 2006),
and ‘Tardibelle’ peaches (Ortiz et al., 2010b), straight-chain esters have
been found to be much more strongly reduced by 1-MCP treatment than
branched-chain esters, suggesting that the biosynthetic pathways that
form these compounds are differently regulated by ethylene action (Raffo
et al., 2009).
With respect to the mechanisms by which volatile production is
decreased by 1-MCP treatment, it has been suggested by several authors
that they are similar to those underlying CA storage (Song et al., 1997;
Mattheis et al., 2005; Song and Forney, 2008). Therefore, in 1-MCP-treated
fruit, the continuous ethylene action necessary to induce higher respiration
rates does not occur, limiting the availability of the substrates required for
maximum aroma volatile biosynthesis. However, other ethylene-­independent
metabolic changes induced by 1-MCP treatment, when applied prior to rip-
ening, might induce changes in substrates and enzymes involved in aroma
biosynthesis (Ferenczi et al., 2006).
Lastly, it must be mentioned that exogenous ethylene is also used
commercially by the fruit industry to induce ripening in bananas, stone
fruit, avocados, melons, kiwifruit, tomatoes, and mangos (Saltveit, 1999).
Although the effect of exogenous ethylene in recovering fruit aroma during
postharvest of fruits, such as apples, is well documented in the research
literature, there is little information available regarding its effect during
commercial application. A study addressing this question was performed
by Stern et al. (1994) using tomatoes, whose results showed that although
ethylene application enhanced aroma by inducing fruit ripening in toma-
toes picked mature green, total volatile production never reached the levels
of those picked table ripe.

16.4.2.4  Other Technologies

Coating fruit with waxes greatly influences aroma production, p ­ articularly
in citrus fruit. This technology is often used to extend the shelf life of fresh
fruits through modification of respiration rates and reduction of water
loss, as well as to prevent postharvest mechanical injury and improve
appearance (Almenar et al., 2009). However, coatings have been found to
induce a high accumulation of fermentative volatiles, such as ethanol and
­acetaldehyde, contributing greatly to the off-flavor of citrus fruit (Baldwin
et al., 1995; Tietel et al., 2010). These unpleasant volatile compounds are
formed under anaerobic conditions that result from a modified internal

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atmosphere within the fruit, in which CO2 is accumulated and O2 is reduced

(Dang et al., 2008). Commercially waxed mandarins and oranges have been
found to be especially prone to the accumulation of high levels of ethanol
(Hagenmaier, 2002; Obenland et al., 2008). It has been suggested that coat-
ing formulations, application procedures, and storage temperatures must
be optimized to maintain the flavor quality of fruits after harvest by attempt-
ing to maximize CO2 and O2 permeability without affecting the acceptable
­characteristics of gloss and low water vapor permeability (Saftner, 1999;
Obenland et al., 2008).
Another postharvest practice that has been reported to influence fruit
aroma is a prestorage calcium treatment. Calcium is widely used in apples
to reduce softening rates and the incidence of some physiological disor-
ders, such as bitter pit. Evidence of the impact of this postharvest practice
on apple aroma has been reported by Ortiz et al. (2009, 2010a), who treated
‘Fuji Kiku-8’ and ‘Golden Reinders’ apples with calcium and showed that
the accumulation of flavor-contributing volatile esters was enhanced after
midterm storage, due to the increased activity of pyruvate decarboxylase
(PDC) and alcohol dehydrogenase (ADH) enzymes.

16.5  Conclusions and Future Perspectives

Currently, the fresh fruit market is expected to provide good-quality ­produce
year-round, which demands a continuous effort by growers and handlers to
maintain the required quality standards of fruits between harvest and con-
sumption. However, efforts to provide fresh fruits of high quality, primarily
in terms of appearance and texture, over a long period of time, often result in
fruits with poor flavor quality, which is currently the main complaint of con-
sumers. Among the flavor quality attributes of fruits, aroma plays a major
role in consumer acceptability. Fruit aroma is greatly affected by growing
conditions, crop management, and marketing operations. As a common
commercial tool within crop management, the early harvest of fruits has
a huge negative impact on fruit aroma. Therefore, the harvest time should
be optimized to balance the requirement of fruit m ­ arketability for long peri-
ods, with the final goal of satisfying the consumer’s desire for a product
with a pleasant taste and aroma. Efforts should also be made to optimize
the postharvest handling practices that have been found to produce detri-
mental effects on fruit aroma, especially those practices that reduce ethyl-
ene biosynthesis or action, or reduce O2 levels in the storage atmosphere.
Taking into account the factors that determine the effects that storage tem-
peratures, storage atmospheres, and other postharvest procedures have on
the aroma quality of fruits would facilitate the selection of the most suitable
storage method for optimizing fruit quality for fresh consumption.

538
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On the other hand, to develop better cultural practices and handling

procedures to improve or maintain fruit aroma, better knowledge of the
mechanisms underlying the changes in the biosynthesis of aroma-related
compounds is required. Although considerable progress has been made
in the elucidation of the metabolic pathways responsible for the s­ ynthesis
of these compounds, information on other enzymes and their regulatory
mechanisms, beyond those covered in this chapter, is still lacking. Further
studies must be directed at discovering the mechanisms associated with
aroma volatile biosynthesis. The combination of this information with the
genetic and genomic resources now available may lead to the generation of
functional molecular markers for use by breeders in selecting for quality
attributes such as taste and aroma. These traits have not been a priority in
breeding programs given that breeders have p ­ rimarily focused on improv-
ing yield, disease resistance, fruit firmness, and shelf life. However, there
is a need to develop a modern breeding approach that is able to additionally
address taste and aroma in order to ultimately improve fruit quality.
In summary, further efforts must be made to generate and integrate
knowledge of fruit genetics and genomics, as well as pre- and postharvest
physiology. Such efforts should address the development of tools for the
fruit industry to use to optimize its practices, with the ultimate aim of pro-
ducing fruits with qualities that are consistent with consumer preferences.

Acknowledgments
The authors thank Conicyt/Fondecyt grants (1060179, 1100273, 1130107)
for funding their studies to further understand flavor metabolism on a­ pricot,
table grape, and avocado.

References
Abdi, N., McGlasson, W.B., Holford, P., Williams, M., and Mizrahi, Y. 1998.
Responses of climacteric and suppressed-climacteric plums to treat-
ment with propylene and 1-methylcyclopropene. Postharvest Biology
and Technology 14, 29–39.
Aharoni, A., and Lewinsohn, E. 2010. Genetic engineering of fruit flavors.
In Y.H. Hui (ed.), Handbook of Fruit and Vegetable Flavors. Wiley,
Hoboken, NJ, 101.
Aharoni, A., Giri, A.P., Verstappen, F.W., Bertea, C.M., Sevenier, R., Sun, Z.,
Jongsma, M.A., Schwab, W., and Bouwmeester, H.J. 2004. Gain and loss
of fruit flavor compounds produced by wild and cultivated strawberry
species. Plant Cell 16, 3110–3131.

539
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Aharoni, A., Keizer, L.C.P., Bouwmeester, H.J., Sun, Z.K., Alvarez-Huerta, M.,
Verhoeven, H.A., Blaas, J., van Houwelingen, A., De Vos, R.C.H., van der
Voet, H., Cansen, R.C., Guis, M., Mol, J., Davis, R.W., Schena, M., van
Tunen, A.J., and O’Connell, A.P. 2000. Identification of the SAAT gene
involved in strawberry flavour biogenesis by use of DNA microarrays.
Plant Cell 12, 647–661.
Almenar, E., Hernández-Muñoz, P., and Gavara, R. 2009. Evolution of selected
volatiles in chitosan-coated strawberries (Fragaria × ananassa) during
refrigerated storage. Journal of Agricultural and Food Chemistry 57,
974–980.
Altisent, R., Echeverría, G., Graell, J., López, L., and Lara, I. 2009.
Lipoxygenase activity is involved in the regeneration of volatile ester-
synthesizing capacity after ultra-low oxygen storage of ‘Fuji’ apple.
Journal of Agricultural and Food Chemistry 57, 4305–4312.
Aubert, C., and Bourger, N. 2004. Investigation of volatiles in Charantais
­cantaloupe melons (Cucumis melo var. cantalupensis): characterization
of aroma constituents in some cultivars. Journal of Agricultural and
Food Chemistry 52, 4522–4528.
Aubert, C., Baumann, S., and Arguel, H. 2005. Optimization of the analysis of
flavor volatile compounds by liquid-liquid microextraction (LLME): appli-
cation to the aroma analysis of melons, peaches, grapes, strawberries,
and tomatoes. Journal of Agricultural and Food Chemistry 53, 8881–8895.
Aubert, C., Günata, Z., Ambid, C., and Baumes, R. 2003. Changes in physico-
chemical characteristics and volatile constituents of yellow- and white-
fleshed nectarines during maturation and artificial ripening. Journal of
Agricultural and Food Chemistry 51, 3083–3091.
Bai, J., Baldwin, E.A., Imahori, Y., Kostenyuk, I., Burns, J., and Brecht, J.K.
2011. Chilling and heating may regulate C6 volatile aroma produc-
tion by different mechanisms in tomato (Solanum lycopersicum) fruit.
Postharvest Biology and Technology 60, 111–120.
Baldwin, E.A., Nisperos-Carriedo, M., Shaw, P.E., and Burns, J.K. 1995. Effect
of coatings and prolonged storage-conditions on fresh orange flavor
­volatiles, degrees Brix, and ascorbic-acid levels. Journal of Agricultural
and Food Chemistry 43, 1321–1331.
Baldwin, E.A., Scott, J.W., Shewmaker, C.K., and Schuch, W. 2000. Flavor
trivia and tomato aroma: Biochemistry and possible mechanisms for
control of important aroma components. HortScience 35, 1013–1022.
Bangerth, F., and Streif, J. 1987. Effect of aminoethoxyvinylglycine and low-
pressure storage on the post-storage production of aroma volatiles by
Golden Delicious apples. Journal of the Science of Food and Agriculture
41, 351–360.
Bangerth, F.K., Song, J., and Streif, J. 2012. Physiological impacts of fruit
­ripening and storage conditions on aroma volatile formation in apple
and strawberry fruit: a review. HortScience 47, 4–10.

540
 F RES H F RUIT AROMA

Bartley, I.M. 1985. Lipid metabolism of ripening apples. Phytochemistry 24,

2857–2859.
Bauchot, A.D., Mottram, D.S., Dodson, A.T., and John, P. 1998. Effect of
aminocyclopropane-1-carboxilic acid oxidase antisense gene on the
formation of volatile esters in cantaloupe Charentais melon. Journal of
Agricultural and Food Chemistry 46, 4787–4792.
Beekwilder, J., Alvarez-Huerta, M., Neef, E., Verstappen, F.W.A., Bowmeester,
H.J., and Aharoni, A. 2004. Functional characterization of enzymes
forming volatile esters from strawberry and banana. Plant Physiology
135, 1865–1878.
Belancic, A., Agosin, E., Ibacache, A., Bordeu, E., Baumes, R., Razungles,
A., and Bayonove, C. 1997. Influence of sun exposure on the aromatic
composition of Chilean Muscat grape cultivars Moscatel de Alejandria
and Moscatel Rosada. American Journal of Enology and Viticulture 48,
181–186.
Bindon, K.A., Dry, P.R., and Loveys, B.R. 2007. Influence of plant water status
on the production of C13-norisoprenoid precursors in Vitis vinifera L.
cv. Cabernet Sauvignon grape berries. Journal of Agricultural and Food
Chemistry 55, 4493–4500.
Blanch, G.P., Flores, G., and Ruiz del Castillo, M.L. 2011. Influence of methyl
jasmonate in conjunction with ethanol on the formation of volatile com-
pounds in berries belonging to the Rosaceae. Postharvest Biology and
Technology 62, 168–178.
Brackmann, A., Streif, J., and Bangerth, F. 1993. Relationship between a
reduced aroma production and lipid metabolism of apples after long-
term controlled-atmosphere storage. Journal of the American Society for
Horticultural Science 118, 243–247.
Brat, P., Yahia, A., Chillet, M., Bugaud, C., Bakry, F., Reynes, M., and Brillouet,
J.M. 2004. Influence of cultivar, growth altitude and maturity stage on
banana volatile compound composition. Fruits 59, 75–82.
Buettner, A., and Schieberle, P. 2001. Evaluation of aroma differences between
hand-squeezed juices from Valencia Late and Navel oranges by quanti-
tation of key odorants and flavor reconstitution experiments. Journal of
Agricultural and Food Chemistry 49, 2387–2394.
Bureau, S., Baumes, R., and Razungles, A. 2000a. Effects of vine or bunch
shading on the glycosylated flavor precursors of Vitis vinifera L. cv.
Syrah. Journal of Agricultural and Food Chemistry 48, 1290–1297.
Bureau, S.M., Razungles, A.J., and Baumes, R.L. 2000b. The aroma of Muscat
of Frontignan grapes: Effect of the light environment of vine or bunch
on volatiles and glycoconjugates. Journal of the Science of Food and
Agriculture 80, 2012–2020.
Cabral, L.M.C., Bravo, A., Freire, M., Jr., Bizzo, H.R., and Matta, V.M. 2010.
Citrus fruits and oranges. In Y.H. Hui (ed.), Handbook of Fruit and
Vegetable Flavors. Wiley, Hoboken, NJ, 265.

541
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Cano-Salazar, J., López, M.L., Crisosto, C.H., and Echeverría, G. 2013. Volatile
compound emissions and sensory attributes of ‘Big Top’ nectarine and
‘Early Rich’ peach fruit in response to a pre-storage treatment before cold
storage and subsequent shelf-life. Postharvest Biology and Technology 76,
152–162.
Chen, G., Hackett, R., Walker, D., Taylor, A., Lin, Z., and Grierson, D. 2004.
Identification of a specific isoform of tomato lipoxygenase (TomloxC)
involved in the generation of fatty acid-derived flavor compounds. Plant
Physiology 136, 2641–2651.
Chuan-qiang, X., Yu-xue, H., and Lin, Y. 2011. Effects of grafting on four
­characteristic ester aromas contents and related enzyme activities in
muskmelon fruit. Northern Horticulture 16, 41–44.
Coetzee, C., and du Toit, W.J. 2012. A comprehensive review on Sauvignon
blanc aroma with a focus on certain positive volatile thiols. Food Research
International 45, 287–298.
Condurso, C., Verzera, A., Dima, G., Tripodi, G., Crinò, P., Paratore, A., and
Romano, D. 2012. Effects of different rootstocks on aroma volatile com-
pounds and carotenoid content of melon fruits. Scientia Horticulturae
148, 9–16.
Contreras, C., and Beaudry, R. 2013. Lipoxygenase-associated apple vola-
tiles and their relationship with aroma perception during ripening.
Postharvest Biology and Technology 82, 28–38.
Crisosto, C.H., Garner, D., Andris, H.L., and Day, K.R. 2004. Controlled
delayed cooling extends peach life. HortTechnology 14, 99–104.
Dandekar, A.M., Teo, G., Defilippi, B.G., Uratsu, S.L., Passey, A.J., Kader,
A.A., Stow, J.R., Colgan, R.J., and James, D.J. 2004. Effect of down-­
regulation of ethylene biosynthesis on fruit flavor complex in apple
fruit. Transgenic Research 13, 373–384.
Dang, K.T., Singh, Z., and Swinny, E.E. 2008. Edible coatings influence fruit
ripening, quality, and aroma biosynthesis in mango fruit. Journal of
Agricultural and Food Chemistry 56, 1361–1370.
DeEll, J.R., Murr, D.P., Mueller, R., Wiley, L., and Porteous, M.D. 2005.
Influence of 1-methylcyclopropene (1-MCP), diphenylamine (DPA), and
CO2 concentration during storage on ‘Empire’ apple quality. Postharvest
Biology and Technology 38, 1–8.
Defilippi, B.G., Dandekar, A.M., and Kader, A.A. 2004. Impact of suppression
of ethylene action or biosynthesis on flavor metabolites in apple (Malus
domestica Borkh) fruits. Journal of Agricultural and Food Chemistry 52,
5694–5701.
Defilippi, B.G., Dandekar, A.M., and Kader, A.A. 2005a. Relationship of ethyl-
ene biosynthesis to volatile production, related enzymes and precursor
availability in apple peel and flesh tissues. Journal of Agricultural and
Food Chemistry 53, 3133–3141.
Defilippi, B.G., Kader, A.A., and Dandekar, A.M. 2005b. Apple aroma: alcohol
acyltransferase, a rate limiting step for ester biosynthesis is regulated
by ethylene. Plant Science 168, 1199–1210.

542
 F RES H F RUIT AROMA

Defilippi, B.G., Manríquez, D., Luengwilai, K., and González-Agüero, M. 2009.

Aroma volatiles: Biosynthesis and mechanisms of modulation d ­ uring
fruit development. Advances in Botanical Research 50, 1–37.
Díaz de León-Sánchez, F., Pelayo-Zaldívar, C., Rivera-Cabrera, F., Ponce-
Valadez, M., Ávila-Alejandre, X., Fernández, F.J., Escalona-Buendía,
H.B., and Pérez-Flores, L.J. 2009. Effect of refrigerated storage on
aroma and alcohol dehydrogenase activity in tomato fruit. Postharvest
Biology and Technology 54, 93–100.
Dixon, J., and Hewett, E.W. 2000. Factors affecting apple aroma/flavour
volatile concentration: a review. New Zealand Journal of Crop and
Horticultural Science 28, 155–173.
Dong, J., Zhang, Y., Tang, X., Jin, W., and Han, Z. 2013. Differences in vola-
tile ester composition between Fragaria × ananassa and F. vesca and
implications for strawberry aroma patterns. Scientia Horticulturae 150,
47–53.
Dudareva, N., Pichersky, E., and Gershenzon, J. 2004. Biochemistry of plant
volatiles. Plant Physiology 135, 1893–1902.
Dunemann, F., Boudichevskaia, A., Grafe, C., Weber, W.E., and Ulrich, D.
2009. QTL and candidate gene mapping for aroma compounds in the
apple progeny ‘Discovery’ × ‘Prima’. Acta Horticulturae 839, 433–440.
Dunemann, F., Ulrich, D., Malysheva-Otto, L., Weber, W.E., Longhi, S.,
Velasco, R., and Costa, F. 2012. Functional allelic diversity of the
apple alcohol acyl-transferase gene MdAAT1 associated with fruit
ester volatile contents in apple cultivars. Molecular Breeding 29,
609–625.
Echeverría, G., Fuentes, T., Graell, J., Lara, I., and López, M.L. 2004a. Aroma
volatile compounds of ‘Fuji’ apples in relation to harvest date and cold
storage technology: a comparison of two seasons. Postharvest Biology
and Technology 32, 29–44.
Echeverría, G., Graell, J., López, M.L., and Lara, I. 2004b. Volatile production,
quality and aroma-related enzyme activities during maturation of ‘Fuji’
apples. Postharvest Biology and Technology 31, 217–227.
El-Sharkawy, I., Manriquez, D., Flores, F.B., Regad, F., Bouzayen, M.,
Latche, A., and Pech, J.C. 2005. Functional characterization of a
melon AAT gene family involved in the biosynthesis of ester v­ olatiles:
Identification of a threonine residue. Plant Molecular Biology 59,
­
345–362.
Escalada, V.S., and Archbold, D.D. 2009. Effects of aminoethoxyvinylglycine
plus 1-methylcyclopropene on ‘Royal Gala’ apple volatile production
after cold storage. HortScience 44, 1390–1394.
Fan, X., and Mattheis, J.P. 1999. Impact of 1-methylcyclopropene and methyl
jasmonate on apple volatile production. Journal of Agricultural and Food
Chemistry 47, 2847–2853.
Fellman, J.K., Mattinson, D.S., Bostick, B.C., Mattheis, J.P., and Patterson,
M.E. 1993. Ester biosynthesis in ‘Rome’ apples subjected to low oxygen
atmospheres. Postharvest Biology and Technology 3, 201–214.

543
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Fellman, J.K., Miller, T.W., Mattinson, D.S., and Mattheis, J.P. 2000. Factors
that influence biosynthesis of volatile flavor compound in apple fruits.
HortScience 35, 1026–1033.
Fellman, J.K., Rudell, D.R., Mattinson, D.S., and Mattheis, J.P. 2003.
Relationship of harvest maturity to flavor regeneration after CA storage
of ‘Delicious’ apples. Postharvest Biology and Technology 27, 39–51.
Ferenczi, A., Song, J., Tian, M., Vlachonasios, K.E., Dilley, D., and Beaudry,
R. 2006. Volatile ester suppression and recovery following 1-methylcy-
clopropene application to apple fruit. Journal of the American Society for
Horticultural Science 131, 691–701.
Ferrandino, A., Carlomagno, A., Baldassarre, S., and Schubert, A. 2012.
Varietal and pre-fermentative volatiles during ripening of Vitis vinifera
cv. Nebbiolo berries from three growing areas. Food Chemistry 135,
2340–2349.
Flores, F., El Yahyaoui, F., De Billerbeck, G., Romojaro, F., Latché, A.,
Bouzayen, M., Pech, J.C., and Ambid, C. 2002. Role of ethylene in the
biosynthetic pathway of aliphatic ester aroma volatiles in Charentais
cantaloupe melons. Journal of Experimental Botany 53, 201–206.
Forney, C.F., Kalt, W., and Jordan, M.A. 2000. The composition of strawberry
aroma is influenced by cultivar, maturity, and storage. HortScience 35,
1022–1026.
Gapper, N.E., McQuinn, R.P., and Giovannoni, J.J. 2013. Molecular and
genetic regulation of fruit ripening. Plant Molecular Biology 82, 575–591.
Golding, J.B., Shearer, D., Wyllie, S.G., and McGlasson, W.B. 1998. Application
of 1-MCP and propylene to identify ethylene-dependent ripening pro-
cesses in mature banana fruit. Postharvest Biology and Technology 12,
87–98.
Gonda, I., Bar, E., Portnoy, V., Lev, S., Burger, J., Schaffer, A.A., Tadmor,
Y., Gepstein, S., Giovannoni, J.J., Katzir, N., and Lewinsohn, E. 2010.
Branched-chain and aromatic amino acid catabolism into aroma volatiles
in Cucumis melo L. fruit. Journal of Experimental Botany 61, 1111–1123.
González-Agüero, M., Troncoso, S., Gudenschwager, O., Campos-Vargas, R.,
Moya-León, A., and Defilippi, B.G. 2009. Differential expression levels of
aroma related genes during ripening of apricot (Prunus armeniaca L.).
Plant Physiology and Biochemistry 47, 435–440.
Hagenmaier, R.D. 2002. The flavor of mandarin hybrids with different
­coatings. Postharvest Biology and Technology 24, 79–87.
Harada, M., Ueda, Y., and Iwata, T. 1985. Purification and some properties of
alcohol acetyltransferase from banana fruit. Plant and Cell Physiology
26, 1067–1074.
Holland, D., Larkov, O., Bar-Ya’akov, I., Bar, E., Zax, A., Brandeis, E., Ravid,
U., and Lewinsohn, E. 2005. Developmental and varietal differences in
volatile ester formation and acetyl-CoA: Alcohol acetyl transferase activ-
ities in apple (Malus × domestica Borkh.) fruit. Journal of Agricultural
and Food Chemistry 53, 7198–7203.

544
 F RES H F RUIT AROMA

Iglesias, I., Echeverría, G., and López, M.L. 2012. Fruit color development,
anthocyanin content, standard quality, volatile compound emissions
and consumer acceptability of several ‘Fuji’ apple strains. Scientia
Horticulturae 137, 138–147.
Imahori, Y., Yamamoto, K., Tanaka, H., and Bai, J. 2013. Residual effects of
low oxygen storage of mature green fruit on ripening processes and
ester biosynthesis during ripening in bananas. Postharvest Biology and
Technology 77, 19–27.
Jayanty, S., Song, J., Rubinstein, N.M., Chong, A., and Beaudry, R.M. 2002.
Temporal relationship between ester biosynthesis and ripening events
in bananas. Journal of the American Society for Horticultural Science 127,
998–1005.
Jouquand, C., and Chandler, C. 2008. A sensory and chemical analysis of
fresh strawberries over harvest dates and seasons reveals factors that
affect eating quality. Journal of the American Society for Horticultural
Science 133, 859–867.
Ke, D., Zhou, L., and Kader, A.A. 1994. Mode of oxygen and carbon dioxide
action on strawberry ester biosynthesis. Journal of the American Society
for Horticultural Science 119, 971–975.
Klee, H.J., and Giovannoni, J.J. 2011. Genetics and control of tomato
fruit  ­ripening and quality attributes. Annual Reviews in Genetics 45,
41–59.
Kondo, S., Setha, S., Rudell, D.R., Buchanan, D.A., and Mattheis, J.P. 2005.
Aroma volatile biosynthesis in apples affected by 1-MCP and methyl
jasmonate. Postharvest Biology and Technology 36, 61–68.
Koundouras, S., Hatzidimitriou, E., Karamolegkou, M., Dimopoulou, E.,
Kallithraka, S., Tsialtas, J.T., Zioziou, E., Nikolaou, N., and Kotseridis,
Y. 2009. Irrigation and rootstock effects on the phenolic concentration
and aroma potential of Vitis vinifera L. cv. Cabernet Sauvignon grapes.
Journal of Agricultural and Food Chemistry 57, 7805–7813.
Langlois, D., Etiévant, P.X., Pierron, P., and Jorrot, A. 1996. Sensory and
instrumental characterisation of commercial tomato varieties. Zeitschrift
für Lebensmittel-Untersuchung und -Forschung 203, 534–540.
Lara, I., Graell, J., López, M.L., and Echeverría, G. 2006. Multivariate analysis
of modifications in biosynthesis of volatile compounds after CA storage
of ‘Fuji’ apples. Postharvest Biology and Technology 39, 19–28.
Larsen, M., and Watkins, C.B. 1995. Firmness and concentrations of acetal-
dehyde, ethyl acetate and ethanol in strawberries stored in controlled
and modified atmospheres. Postharvest Biology and Technology 5,
39–50.
Lewinsohn, E., Schalechet, F., Wilkinson, J., Matsui, K., Tadmor, Y., Nam,
K.H., Amar, O., Lastochkin, E., Larkov, O., Ravid, U., Hiatt, W., Gepstein,
S., and Pichersky, E. 2001. Enhanced levels of the aroma and flavor com-
pound S-linalool by metabolic engineering of the terpenoid pathway in
tomato fruits. Plant Physiology 127, 1256–1265.

545
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Lewinsohn, E., Sitrit, Y., Bar, E., Azulay, Y., Meir, A., Zamir, D., and Tadmor,
Y. 2005. Carotenoid pigmentation affects the volatile composition of
tomato and watermelon fruits, as revealed by comparative genetic anal-
yses. Journal of Agricultural and Food Chemistry 53, 3142–3148.
Li, D., Xu, Y., Xu, G., Gu, L., Li, D., and Shu, H. 2006. Molecular cloning and
expression of a gene encoding alcohol acyltransferase (MdAAT2) from
apple (cv. Golden Delicious). Phytochemistry 67, 658–667.
Lin, D., Huang, D., and Wang, S. 2004. Effects of potassium levels on fruit
quality of muskmelon in soilless medium culture. Scientia Horticulturae
102, 53–60.
Linsenmeier, A.W., and Lohnertz, O. 2007. Changes in norisoprenoid levels
with long-term nitrogen fertilisation in different vintages of Vitis vinif-
era var. Riesling wines. South African Journal of Enology and Viticulture
28, 17–24.
Longhurst, T.J., Tung, H.F., and Brady, C.J. 1990. Developmental regulation
of the expression of alcohol dehydrogenase in ripening tomato fruits.
Journal of Food Biochemistry 14, 421–433.
Luan, F., Mosandl, A., Münch, A., and Wüst, M. 2005. Metabolism of ­geraniol
in grape berry mesocarp of Vitis vinifera L. cv. Scheurebe: demon-
stration of stereoselective reduction, E/Z isomerization, oxidation.
Phytochemistry 66, 295–303.
Manríquez, D., El-Sharkawy, I., Flores, F.B., El-Yahyaoui, F., Regad, F.,
Bouzayen, M., Latche, A., and Pech, J.C. 2006. Two highly divergent
alcohol dehydrogenases of melon exhibit fruit ripening-specific expres-
sion and distinct biochemical characteristics. Plant Molecular Biology
61, 675–685.
Mattheis, J.P., and Fellman, J.K. 1999. Pre-harvest factors influencing flavor
of fresh fruit and vegetables. Postharvest Biology and Technology 15,
227–232.
Mattheis, J.P., Fan, X., and Argenta, L.C. 2005. Interactive responses of Gala
apple fruit volatile production to controlled atmosphere storage and
chemical inhibition of ethylene action. Journal of Agricultural and Food
Chemistry 53, 4510–4516.
Ménager, I., Jost, M., and Aubert, C. 2004. Changes in physicochemical
characteristics and volatile constituents of strawberry (cv. Cigaline)
during maturation. Journal of Agricultural and Food Chemistry 52,
1248–1254.
Miller, T.W., Fellman, J.K., Mattheis, J.P., and Mattinson, D.S. 1998. Factors
that influence volatile ester biosynthesis in ‘Delicious’ apples. Acta
Horticulturae 464, 195–200.
Mir, N.A., Perez, R., Schwallier, P., and Beaudry, R. 1999. Relationship
between ethylene response manipulation and volatile production in
Jonagold variety apples. Journal of Agricultural and Food Chemistry 47,
2653–2659.

546
 F RES H F RUIT AROMA

Moya-León, M.A., Vergara, M., Bravo, C., Montes, M.E., and Moggia, C. 2006.
1-MCP treatment preserves aroma quality of ‘Packham's Triumph’
pears during long-term storage. Postharvest Biology and Technology 42,
185–197.
Obando-Ulloa, J.M., Moreno, E., García-Mas, J., Lammertyn, J., Monforte,
A.J., and Fernández-Trujillo, J.P. 2008. Climacteric or non-climacteric
behavior in melon fruit. 1. Aroma volatiles. Postharvest Biology and
Technology 49, 27–37.
Obenland, D., Collin, S., Sievert, J., and Arpaia M.L. 2013. Mandarin flavor
and aroma volatile composition are strongly influenced by holding tem-
perature. Postharvest Biology and Technology 82, 6–14.
Obenland, D., Collin, S., Sievert, J., Fjeld, K., Doctor, J., and Arpaia, M.L.
2008. Commercial packing and storage of Navel oranges alters aroma
volatiles and reduces flavor quality. Postharvest Biology and Technology
47, 159–167.
Oeller, P.W., Min-Wong, L., Taylor, L.P., Pike, D.A., and Theologis, A. 1991.
Reversible inhibition of tomato fruit senescence by antisense RNA.
Science 254, 437–439.
Ojeda-Real, L.A., Lobit, P., Cárdenas-Navarro, R., Grageda-Cabrera, O.,
Farías-Rodríguez, R., Valencia-Cantero, E., and Macías-Rodríguez, L.
2009. Effect of nitrogen fertilization on quality markers of strawberry
(Fragaria × ananassa Duch. cv. Aromas). Journal of the Science of Food
and Agriculture 89, 935–939.
Ortiz, A., Echeverría, G., Graell, J., and Lara, I. 2009. Calcium dips enhance vol-
atile emission of cold-stored ‘Fuji Kiku-8’ apples. Journal of Agricultural
and Food Chemistry 57, 4931–4938.
Ortiz, A., Echeverría, G., Graell, J., and Lara, I. 2010a. The emission of flavour-
contributing volatile esters by ‘Golden Reinders’ apples is improved
after mid-term storage by postharvest calcium treatment. Postharvest
Biology and Technology 57, 114–123.
Ortiz, A., Graell, J., López, M.L., Echeverría, G., and Lara, I. 2010b. Volatile
ester-synthesising capacity in ‘Tardibelle’ peach fruit in response to
controlled atmosphere and 1-MCP treatment. Food Chemistry 123,
698–704.
Osorio, S., Muñoz, C., and Valpuesta, V. 2010. Physiology and biochemistry of
fruit flavors. In Y.H. Hui (ed.), Handbook of Fruit and Vegetable Flavors.
Wiley, Hoboken, NJ, 129.
Paillard, N.M.M. 1986. Evolution of the capacity of aldehyde production
by crushed apple tissues, during an extended storage of fruits. In G.
Charalambous (ed.), The Shelf Life of Foods and Beverages. Elsevier
Science, Amsterdam, 369.
Pelayo, C., Ebeler, S.E., and Kader, A.A. 2003. Postharvest life and flavor qual-
ity of three strawberry cultivars kept at 5°C in air or air + 20 kPa CO2.
Postharvest Biology and Technology 27, 171–183.

547
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Pelayo-Zaldívar, C., Abda, J.B., Ebeler, S.E., and Kader, A.A. 2007. Quality
and chemical changes associated with flavor of ‘Camarosa’ straw-
berries in response to a CO2-enriched atmosphere. HortScience 42,
299–303.
Pichersky, E., Noel, J.P., and Dudareva, N. 2006. Biosynthesis of plant
­volatiles: nature’s diversity and ingenuity. Science 311, 808–811.
Plotto, A., McDaniel, M., and Mattheis, J. 2000. Characterization of changes
in ‘Gala’ apple aroma during storage using Osme analysis, a gas chro-
matography-olfactometry technique. Journal of the American Society for
Horticultural Science 125, 714–722.
Raffo, A., Kelderer, M., Paoletti, F., and Zanella, A. 2009. Impact of innova-
tive controlled atmosphere storage technologies and postharvest treat-
ments on volatile compound production in cv. Pinova apples. Journal of
Agricultural and Food Chemistry 57, 915–923.
Reid, S.J., Watkins, C.B., Janssen, B. and Ross, G.S. 1996. An alcohol dehy-
drogenase cDNA clone (accession no. Z48234) isolated from apple fruit
(PGR96-043). Plant Physiology 111, 947.
Rizzolo, A., Cambiaghi, P., Grassi, M., and Zerbini, P.E. 2005. Influence of
1-methylcyclopropene and storage atmosphere on changes in volatile
compounds and fruit quality of conference pears. Journal of Agricultural
and Food Chemistry 53, 9781–9789.
Roscher, R., Bringmann, G., Schreier, P., and Schwab, W. 1998. Radiotracer
studies on the formation of 2,5-dimethyl-4-hydroxy-3(2H)-furanone in
detached ripening strawberry fruits. Journal of Agricultural and Food
Chemistry 46, 1488–1493.
Rouseff, R.L., Ruiz Perez-Cacho, P., and Jabalpurwala, F. 2009. Historical
review of citrus flavor research during the past 100 years. Journal of
Agricultural and Food Chemistry 57, 8115–8124.
Saftner, R.A. 1999. The potential of fruit coating and film treatments for
improving the storage and shelf-life qualities of ‘Gala’ and ‘Golden
Delicious’ apples. Journal of the American Society for Horticultural
Science 124, 682–689.
Saltveit, M.E. 1999. Effect of ethylene on quality of fresh fruits and vegeta-
bles. Postharvest Biology and Technology 15, 279–292.
Sanz, C., and Pérez, A.G. 2010. Plant metabolic pathways and flavor biosyn-
thesis. In Y.H. Hui (ed.), Handbook of Fruit and Vegetable Flavors. Wiley,
Hoboken, NJ, 129.
Sanz, C., Olías, J.M., and Pérez, A.G. 1997. Aroma biochemistry of fruits
and vegetables. In F.A. Tomas-Barberan and R.J. Robins (eds.),
Phytochemistry of Fruits and Vegetables. Clarendon Press, Oxford, 125.
Schaffer, R.J., Friel, E.N., Souleyre, E.J.F., Bolitho, K., Thodey, K., Ledger,
S., Bowen, J.H., Ma, J.H., Nain, B., Cohen, D., Gleave, A.P., Crowhurst,
R.N., Janssen, B.J., Yao, J.L., and Newcomb, R.D. 2007. A genomics
approach reveals that aroma production in apple is controlled by ethyl-
ene predominantly at the final step in each biosynthetic pathway. Plant
Physiology 144, 1899–1912.

548
 F RES H F RUIT AROMA

Schuhr, C.A., Radykewicz, T., Sagner, S., Latzel, C., Zenk, M.H., Arigoni, D.,
Bacher, A., Rohdich, F., and Eisenreich, W. 2003. Quantitative assess-
ment of crosstalk between the two isoprenoid biosynthesis pathways in
plants by NMR spectroscopy. Phytochemistry Reviews 2, 3–16.
Schultz, H.R., Löhnertz, O., Bettner, W., Bálo, B., Linsenmeier, A., Jähnisch,
A., Müller, M., Gaubatz, B., and Varadi, G. 1998. Is grape composition
affected by current levels of UV-B radiation? Vitis 37, 191–192.
Schwab, W. 2013. Natural 4-hydroxy-2,5-dimethyl-3(2H)-furanone (Furaneol®).
Molecules 18, 6936–6951.
Schwab, W., Davidovich-Rikanati, R., and Lewinsohn, E. 2008. Biosynthesis of
plant-derived flavor compounds. Plant Journal 54, 712–732.
Shalit, M., Katzir, N., Tadmor, Y., Larkov, O., Burger, Y., Shalekhet, F.,
Lastochkin, E., Ravid, U., Amar, O., Edelstein, M., Karchi, Z., and
Lewinsohn, E. 2001. Acetyl Co-A: Alcohol acetyltransferase activity and
aroma formation in ripening melon fruits. Journal of Agricultural and
Food Chemistry 49, 794–799.
Shan, W., Zhao, C., Fan, J., Cong, H., Liang, S., and Yu, X. 2012. Antisense sup-
pression of alcohol acetyltransferase gene in ripening melon fruit alters
volatile composition. Scientia Horticulturae 139, 96–101.
Song, J., and Bangerth, F. 1994. Production and development of volatile
aroma compounds of apple fruits at different times of maturity. Acta
Horticulturae 368, 150–157.
Song, J., and Bangerth, F. 1996. The effect of harvest date and aroma com-
pound production from ‘Golden Delicious’ apple fruit and relationship to
respiration and ethylene production. Postharvest Biology and Technology
8, 259–269.
Song, J., and Bangerth, F. 2003. Fatty acids as precursors for aroma volatile
biosynthesis in pre-climacteric and climacteric apple fruit. Postharvest
Biology and Technology 30, 113–121.
Song, J., and Forney, C.F. 2008. Flavour volatile production and regulation in
fruit. Canadian Journal of Plant Science 88, 537–550.
Song, J., Tian, M.S., Dilley, D.R., and Beaudry, R.M. 1997. Effect of 1-MCP
on apple fruit ripening and volatile production. HortScience 32, 536–541.
Speirs, J., Lee, E., Holt, K., Yong-Duk, K., Steele Scott, N., Loveys, B., and
Schuch, W. 1998. Genetic manipulation of alcohol dehydrogenase levels
in ripening tomato fruit affects the balance of some flavor aldehydes and
alcohols. Plant Physiology 117, 1047–1058.
Stern, D.J., Buttery, R.G., Teranishi, R., Ling, L., Scott, K., and Cantwell, M.
1994. Effect of storage and ripening on fresh tomato quality, part I. Food
Chemistry 49, 225–231.
Streif, J., and Bangerth, F. 1988. Production of volatile aroma substances by
‘Golden Delicious’ apple fruits after storage for various times in different
CO2 and O2 concentrations. Journal of Horticulture Science 63, 193–199.
Tesniere, C., and Verries, C. 2000. Molecular cloning and expression of
cDNAs encoding alcohol dehydrogenases from Vitis vinifera L. during
berry development. Plant Science 157, 77–88.

549
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Tietel, Z., Bar, E., Lewinsohn, E., Feldmesser, E., and Fallik, E. 2010. Effects
of wax coatings and postharvest storage on sensory quality and aroma
volatiles composition of ‘Mor’ mandarins. Journal of the Science of Food
and Agriculture 90, 995–1007.
Tietel, Z., Porat, R., Weiss, K., and Ulrich, D. 2011. Identification of aroma-
active compounds in fresh and stored ‘Mor’ mandarins. International
Journal of Food Science and Technology 46, 2225–2231.
Tikunov, Y., Lommen, A., de Vos, C.H.R., Verhoeven, H.A., Bino, R.J., Hall,
R.D., and Bovy, A.G. 2005. A novel approach for nontargeted data analy-
sis for metabolomics: Large-scale profiling of tomato fruit volatiles.
Plant Physiology 139, 1125–1137.
Tokitomo, Y., Steinhaus, M., Büttner, A., and Schieberle, P. 2005. Odor-active
constituents in fresh pineapple (Ananas comosus [L.] Merr.) by quantita-
tive and sensory evaluation. Bioscience, Biotechnology, and Biochemistry
69, 1323–1330.
Vandendriessche, T., Geerts, P., Membrebe, B.N., Keulemans, J., Nicolaï,
B.M., and Hertog, M.L.A.T.M. 2013. Journeys through aroma space: A
novel approach towards the selection of aroma-enriched strawberry cul-
tivars in breeding programmes. Plant Breeding 132, 217–223.
Veit-Köhler, U., Krumbein, A., and Kosegarten, H. 1999. Effect of different
water supply on plant growth and fruit quality of Lycopersicon esculen-
tum. Journal of Plant Nutrition and Soil Science 162, 583–588.
Verzera, A., Dima, G., Tripodi, G., Ziino, M., Lanza, C.M., and Mazzaglia, A.
2011. Fast quantitative determination of aroma volatile constituents
in melon fruits by headspace-solid-phase microextraction and gas
chromatography–mass spectrometr y. Food Analytical Methods 4,
141–149.
Wang, J., and DeLuca, V. 2005. The biosynthesis and regulation of biosynthe-
sis of Concord grape fruit esters, including ‘Foxy’ methylanthranilate.
Plant Journal 44, 606–619.
Wang, Y., Yang, C., Li, S., Yang, L., Wang, Y., Zhao, J., and Jiang, Q. 2009.
Volatile characteristics of 50 peaches and nectarines evaluated by HP–
SPME with GC–MS. Food Chemistry 116, 356–364.
Watson, R., Wright, C.J., McBurney, T., Taylor, A.J., and Linforth, R.S.T.
2002. Influence of harvest date and light integral on the development
of strawberry flavour compounds. Journal of Experimental Botany 53,
2121–2129.
Wein, M., Lavid, N., Lunkenbein, S., Lewinsohn, E., Schwab, W., and
Kaldenhoff, R. 2002. Isolation, cloning and expression of a multifunc-
tional O-methyltransferase capable of forming 2,5-dimethyl-4-methoxy-
3(2H)-furanone, one of the key aroma compounds in strawberry fruits.
Plant Journal 31, 755–765.
Wendakoon, S.K., Ueda, Y., Imahori, Y., and Ishimaru, M. 2006. Effect of
short-term anaerobic conditions on the production of volatiles, activity
of alcohol acetyltransferase and other quality traits of ripened bananas.
Journal of the Science of Food and Agriculture 86, 1475–1480.

550
 F RES H F RUIT AROMA

Wyllie, G., and Fellman, J.K. 2000. Formation of volatile branched chain
esters in banana (Musa sapientum L). Journal of Agricultural and Food
Chemistry 48, 3493–3496.
Xi, W., Zhang, B., Shen, J., Sun, C., Xu, C., and Chen, K. 2012. Intermittent
warming alleviated the loss of peach fruit aroma-related esters by regu-
lation of AAT during cold storage. Postharvest Biology and Technology
74, 42–48.
Yang, C., Wang, Y., Wu, B., Fang, J., and Li, S. 2011. Volatile compounds evo-
lution of three table grapes with different flavour during and after matu-
ration. Food Chemistry 128, 823–830.
Zabetakis, I., and Holden, M.A. 1997. Strawberry flavour: Analysis and bio-
synthesis. Journal of the Science of Food and Agriculture 74, 421–434.
Zhang, B., Shen, J.Y., Wei, W.W., Xi, W.P., Xu, C.J., Ferguson, I., and Chen,
K.S. 2010. Expression of genes associated with aroma formation
derived from the fatty acid pathway during peach fruit ripening. Journal
of Agricultural and Food Chemistry 58, 6157–6165.
Zhang, B., Xi, W.P., Wei, W.W., Shen, J.Y., Ferguson, I., and Chen, K.S. 2011.
Changes in aroma-related volatiles and gene expression during low
temperature storage and subsequent shelf-life of peach fruit. Postharvest
Biology and Technology 60, 7–16.
Zhang, B., Yin, X.R., Shen, J.Y., Chen, K.S., and Ferguson, I.B. 2009. Volatiles
production and lipoxygenase gene expression in kiwifruit peel and flesh
during fruit ripening. Journal of the American Society for Horticultural
Science 134, 472–477.
Zhu, H.L., Zhu, B.Z., Fu, D.Q., Xie, Y.H., Hao, Y.L., and Luo, Y.B. 2005. Role
of ethylene in the biosynthetic pathways of aroma volatiles in ripening
fruit. Russian Journal of Plant Physiology 52, 691–695.
Zhu, Y., Rudell, D.R., and Mattheis, J.P. 2008. Characterization of cultivar
differences in alcohol acyltransferase and 1-aminocyclopropane-1-­
carboxylate synthase gene expression and volatile ester emission
during apple fruit maturation and ripening. Postharvest Biology and
Technology 49, 330–339.
Zorrilla-Fontanesi, Y., Rambla, J.L., Cabeza, A., Medina, J.J., Sánchez-
Sevilla, J.F., Valpuesta, V., Botella, M.A., Granell, A., and Amaya, I.
2012. Genetic analysis of strawberry fruit aroma and identification of
O-methyltransferase FaOMT as the locus controlling natural variation in
mesifurane content. Plant Physiology 159, 851–870.

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Chapter 17

Flavor and Aroma

Compounds of Some
Exotic Tropical
Fruits and Berries:
Biosynthetic Pathways
and Metabolism
Ola Lasekan
University Putra Malaysia, Serdang, Malaysia

Abstract 554
17.1 Introduction 555
17.2 Exotic Fruits and Their Flavor Profiles 556
17.2.1  Lychee (Litchi chinensis) 556
17.2.2  Rambutan (Nephelium lappaceum L.) 556
17.2.3  Yellow Passion (Passiflora edulis) 558
17.2.4  Durian Fruit (Durio zibethinus) 559
17.2.5  Star Fruit or Carambola (Averrhoa
carambola L.) 560
17.2.6 Mangosteen (Garcinia mangostana) 560

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17.2.7    Snake Fruit (Salacca edulis Reinw) 560

17.2.8    Costa Rican Guava (Psidium friedrichsthalium) 565
17.2.9    Pitanga Fruit (Eugenia uniflora L.) 566
17.2.10  Umbu-Caja Fruit (Spondias citherea) 566
17.2.11  Camu-Camu Fruit (Myrciaria dubia) 566
17.2.12  Cupuacu Fruit (Theobroma grandiflorum) 567
17.2.13  Araca-Boi Fruit (Eugenia stipitata) 567
17.2.14  Mangaba Fruit (Hancornia speciosa Gomes) 567
17.2.15  Garcinia Fruit (Garcinia dulcis Kurz) 568
17.2.16  Guabiju Fruit (Myrcianthes pungens Berg) 568
17.2.17  Guabiroba Fruit (Campomanesia
xanthocarpa Berg) 568
17.2.18  Bacuri Fruit (Platonia sculenta) 568
17.2.19  Cashew Fruit (Anacardium occidentale L.) 569
17.2.20  Melon Fruit (Cucumis melo) 569
17.2.21  Jackfruit (Artocarpus heterophyllus Lam.) 570
17.2.22  Sapodilla Fruit (Achras sapota L.) 570
17.2.23  Genipap Fruit (Genipa americana) 571
17.2.24  Soursop Fruit (Annona muricata) 571
17.2.25  Acerola Fruit (Malphigia glabra L.) 572
17.2.26  Tamarind Fruit (Tamarindus indica L.) 572
17.2.27  Velvet Tamarind (Dialium guineense) 572
17.2.28  African Star Apple Fruit (Chrysophillum
albidum) 573
17.3 Aroma Compounds’ Biosynthetic Pathways and
Metabolism 573
17.3.1  Fruit’s Volatile Ester Metabolism 573
17.3.2  Fruit’s Volatile Terpenoid Metabolism 575
17.3.3  Sulfur Volatile Compounds’ Biosynthetic
Pathway 577
17.4 Conclusions and Future Perspectives 579
References 579

Abstract
The characteristic flavor of exotic tropical fruits is one of their most attrac-
tive attributes to consumers. In this chapter, the enormous diversity of
exotic fruit flavors is reviewed. Classifying some of the exotic fruits into two
classes on the basis of whether esters or terpenes predominate in the aroma
was also attempted. Indeed, as far as exotic tropical fruits are concerned,
the majority of fruits have terpenes predominating in their aroma profile.

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Some of the fruits in this group are the Amazonian fruits, such as pitanga,
umbu-caja, camu-camu, garcinia, and bacuri. The ester group is made up
of rambutan, durians, star fruit, snake fruit, acerola, tamarind, sapodilla,
genipap, soursop, cashew, melon, jackfruit, and cupuacu. Also, this review
summarizes recent progress in the characterization of esters, terpenoids,
and sulfur volatile compounds’ biosynthetic genes and their spatiotemporal
expression patterns.

17.1 Introduction
Among the many attractive attributes that create demand for fruits from
the tropics and subtropics is the noticeable flavor characteristic. In addi-
tion, these fruits are often inexpensive and extremely rich in vitamins and
can be used in a wide range of food products. A great deal of attention has
been directed toward the volatile compositions of a wide variety of fruits
over the past several decades. In a few cases, individual “character-impact
compounds” have been pinpointed as being responsible for a characteris-
tic flavor, but most fruit flavors seem to be due to the integrated response
to a large number of contributing compounds (Marostica and Pastore,
2007). Indeed, as far as exotic tropical fruits are concerned, a scan of the
literature (Van Straten et al., 1988) shows that esters and terpenes are
their most prolific volatile components. It would also appear that a rough
classification of such fruit can be made on the basis of whether esters or
terpenes predominate in the aroma. In very rare cases, other classes of
compounds contribute significantly to the aroma notes of these exotic
fruits. For example, the attractive tropical flavor note of ripe yellow pas-
sion fruit has been shown to be associated with trace levels of sulfur vola-
tiles (Werkhoff et al., 1998). Volatile sulfur compounds are important trace
constituents of natural products and play an important role in the sensory
properties of food flavors.
The characteristic flavor of exotic fruits is one of the most attractive
attributes to consumers. The enormous diversity of exotic fruits represents
a promising area for research on aromas, with unusual sensory properties.
Nowadays, food industries are looking at how to use these volatiles to pro-
duce amazing new products that can accommodate this demand (Marostica
and Pastore, 2007). Accordingly, more research should be done in parallel
with the increasing demand of the food flavor industries for these exotic
fruits. The outcomes of this research may be necessary to stimulate the
production of accurate and natural flavor compounds for use in different
food systems. In view of the above, this work reviews the distinctive flavor
constituents of some exotic tropical fruits that are not well known in the
Western world.

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17.2  Exotic Fruits and Their Flavor Profiles

17.2.1 Lychee (Litchi chinensis)

The aroma of lychee (Litchi chinensis) is often described as being rose-
floral and citruslike (Ong and Acree, 1998). Although lychee, a member
of the Sapindaceae family, is a subtropical fruit native to China, its com-
mercial importance and popularity among consumers have continued to
expand to markets outside Asia (Klotzbach, 1995). Johnston et al. (1980)
were the first to report on the volatiles of lychee, suggesting the citrus
flavor was due to the presence of compounds such as geranial and neral,
and its floral note was due mainly to 2-phenyl ethanol. Two other stud-
ies (Toulemonde and Beauverd, 1985; Froehlich and Schreier, 1986) on
the headspace and neutral volatiles in this fruit identified limonene, rose
oxide, nonanal, decanal, citronellol, and geraniol as significant contribu-
tors to the fruity-floral and citrus notes, respectively. A gas chromatogra-
phy–olfactometry (GC-O) analysis of the fruit by Ong and Acree (1998)
detected about 60 odor-active volatiles in the fruit extract. Among the com-
pounds that had significant odor activity were geraniol, guaiacol, vanillin,
2-acetyl-2-thiazoline, 2-phenyl ethanol, (Z)-2-nonenal, β-damascenone,
1-octen-3-ol, furaneol, linalool, (E)-2-nonenal, isobutyl acetate, cis-rose
oxide, and isovaleric acid. Ong and Acree (1998) confirmed that 2-­phenyl
ethanol was probably responsible for the flora character, and that the cit-
rus-fruity aroma is due to the presence of many odor-active terpenes, par-
ticularly geraniol (Table 17.1).

17.2.2 Rambutan (Nephelium lappaceum L.)

Another member of the Sapindaceae family is the rambutan, with charac-
teristic fruity-sweet and fatty-green aroma notes (Ong and Acree, 1998).
Isolation and characterization of odor-active compounds in rambutan by
GC-O and gas chromatography–mass spectrometry produced the 20 most
potent odorants: β-damascenone, (E)-4,5-epoxy-(E)-2-decenal, vanillin,
(E)-2-nonenal, phenyl acetic acid, cinnamic acid, ethyl-2-methyl butyrate,
furaneol, m-cresol, guaiacol, nonanal, 2-phenyl ethanol, heptanoic acid,
maltol, 2,6-nonadienal, (Z)-2-nonenal, γ-nonalactone, 3-phenyl propionic
acid, δ-decalactone and one unknown compound. Further analysis revealed
that β-damascenone, ethyl-2-methyl butyrate, 2,6-nonadienal, (E)-2-nonenal,
and nonanal were the main contributors to the fruity aroma.

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Table 17.1  Major Flavor Compounds in Some Exotic Tropical Fruits

Compound Structure Odor and Occurrence
β-Damascenone O Fruity, sweet
Lychee, rambutan,
yellow passion,
garcinia, star fruits
cis-Rose Flowery, roselike
Lychee

O
cis-Hex-3-enyl Fruity, sweet
acetate O Mangosteen, star fruit

O
Ethyl butyrate O Fruity
Cupuacu, Costa Rican
O guava, durian,
camu-camu
Ethyl hexanoate O Fruity
Cupuacu, Costa Rican
guava, yellow passion
O
fruit, durian
Geraniol Roselike
OH
Lychee, star fruit,
mangaba fruit,
garcinia, evodia,
yellow passion
Geranyl acetol OH Sweet, fruity
Yellow passion fruit

Hexyl acetate O Fruity

Isobutyl acetate
Mangaba, mangosteen
O
O Fruity, cherry
Lychee, rambutan
O

(Continued )

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Table 17.1  Major Flavor Compounds in Some Exotic Tropical Fruits

Compound Structure Odor and Occurrence
Megastigma Roselike, raspberry-like
4,6,8-trienes Star fruit, yellow passion
fruit, purple passion
fruit
cis- and Fruity
trans-Ocimene Pitanga, umbu-caja,
araca, lychee, garcinia,
longan, evodia, Costa
Rican guava, yellow
H3C CH3 H3C CH3 passion

cis trans
β-Pinene Woody-green
Garcinia, Costa Rican
guava, yellow passion,
pitanga, guabiroba,
araca-boi, umbu-caja,
camu-camu

(Z)-5-Tangerinol Sweet, flora

Yellow passion, purple
O passion
O

β-Selinene Fruity
Costa Rican guava,
guabiroba, pitanga,
araca

17.2.3  Yellow Passion (Passiflora edulis)

One of the most popular and best-known tropical fruits having a floral, ester
aroma with an exotic sulfury note is the yellow passion fruit. The attractive
flavor note of ripe yellow passion fruit has been shown to be associated with
trace levels of sulfur volatiles (Werkhoff et al., 1998). Volatile sulfur com-
ponents are important trace constituents of natural products and play an
important role in the sensory properties of food flavors. Sulfur-containing
components combine high odor intensities and low threshold values and
have been identified as character-impact substances in various foods and

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beverages (Mussinan and Keelan, 1994). The analysis of the flavor com-
position of yellow passion fruit by four different isolation techniques,
namely, (1) vacuum headspace sampling (VHS), (2) the dynamic head-
space method, (3) simultaneous distillation and extraction at atmospheric
pressure, and (4) simultaneous distillation and extraction under reduced
pressure, produced approximately 180 flavor components (Werkhoff et al.,
1998). While the flavor of yellow passion fruit is determined by numer-
ous volatile components, the attractive tropical flavor note of ripe yellow
passion fruit is mainly attributed to the presence of trace levels of sulfur
volatiles in combination with other compounds. Compounds contribut-
ing significantly to the fruit aroma note are cis- and trans-γ-jasmine lac-
tone; δ-jasmine lactone, with a sweet, creamy, and fruity-flowery note;
trans-marmelolactone, with a strong fruity, floral, and quincelike aroma
(Tsuneya et al., 1980); cycloionone, with a fruity, sweet, floral, and woody
aroma (Nijssen et al., 1996); 4-hydroxy-2,5-dimethyl-3(2H)-furanone and
4-methoxy-2,5-dimethyl-3(2H)-furanone, with their characteristic fruity
notes; furaneol acetate; 3-mercaptohexyl pentanoate; 3-(methylthiol)pro-
pyl acetate, methionyl acetate, and geranyl acetol, with a sweet, fruity note;
and neryl acetol and (Z)-5-tangerinol, with sweet floral and woody notes,
respectively (Table 17.1).

17.2.4  Durian Fruit (Durio zibethinus)

Durian, a member of the Bombaceae family, is one of the most well-known
tropical fruits in Southeast Asia. It has distinct onionlike and fruity notes
(Chin et al., 2008). Studies on the volatile constituents of durian have
revealed a total of 137 volatile compounds (Naf and Velluz, 1996; Weenen
et al., 1996; Jaswir et al., 2005). These compounds are composed mostly of
esters and acids. Among various durian varieties, sulfur-containing vola-
tiles such as thiols, disulfides, and trisulfides were reported as the major
volatile constituents contributing to the strong onionlike note (Weenen
et al., 1996). A study (Chin et al., 2008) on the analysis of the volatile com-
pounds of durian varieties, using headspace solid-phase microextraction
(SPME) coupled with fast gas chromatography–mass spectrometry (GC-
MS), identified a total of 39 volatile compounds. Those compounds that
contributed significantly to the durian flavor were listed as ethyl butano-
ate, ethyl hexanoate, methyl hexanoate, methyl butanoate, ethyl-3-methyl
butanoate, propyl hexanoate, methyl octanoate, diethyl trisulfide, methyl
propyl disulfide, ethyl propyl disulfide, dipropyl disulfide, disulfide, trithi-
olanes, acetate, and s-propyl thioacetate. While the sulfur volatiles were
responsible for the strong garlic, onionlike odor, the esters contributed sig-
nificantly to the fruity note.

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17.2.5  Star Fruit or Carambola (Averrhoa carambola L.)

This is an extremely attractive fruit with a unique ribbed stellate shape (Table
17.2) and a distinct apple, peach, plumlike aroma (MacLeod and Ames, 1990).
The volatile components of star fruit have been reported by a few groups of
researchers (Toulemonde and Beauverd, 1985; Willson et al., 1985; MacLeod
and Ames, 1990). A total of 153 volatile compounds have been characterized
to date, of which 64 are esters. Many of these are methyl and ethyl esters
of alkanoic, alkenoic, alkadioic, and benzenoid acids, but also included are
some propyl, butyl, pentyl, hexyl, hexenyl, and octyl esters (MacLeod and
Ames, 1990). Some of the compounds that contributed significantly to the
star fruit aroma are hexyl methyl propanoate, hex-2(E)-enyl butanoate, hex-
3(Z)-enyl pentanoate, hex-3(Z)-enyl hexanoate, hexyl hexanoate, hex-2(E)-
enyl hexanoate, hex-2(E)-enyl heptanoate, and hex-2(E)-enyl propanoate,
all of which are metabolically related in that they are derived from the C6
alcohols, hexan-1-ol, hex-3(Z)-en-1-ol, and hex-2(E)-en-1-ol, the well-known
products of lipoxygenase oxidation of 18:2 and 18:3 fatty acids (Enzell, 1981).
Other compounds are γ-decalactone, γ-undecalactone, δ-dodecalactone,
δ-dodecalactone, δ-tridecalactone, and δ-undecalactone. Indeed, the C10
and C12 lactones, in particular, are well known as important contributors to
the peachlike odor quality (Buttery, 1981). Also of importance are β-ionone,
β-damascenone, four isomeric megastigma-4,6,8-trienes (Table 17.1), and
megastigma-5,8(E)- and -(Z)-dien-4-one.

17.2.6 Mangosteen (Garcinia mangostana)

The mangosteen is generally admitted to be one of the most attractive of tropi-
cal fruits (Table 17.2). It has a characteristic sweet cum glutinous impression
(MacLeod and Pieris, 1982). A gas chromatography–mass spectrometry
analysis of the fruit identified 28 volatile compounds. These comprised eight
sesquiterpenes: α-terpineol (with pleasant aroma similar to lilac) and delta-
cadinene α-copaene (buttery aroma note), guaiene, α-bisabolene (dull fruity),
valencene (caramelized orange aroma), δ-cadmene (nutty, fatty), γ-cadinene
(fruity), hexyl acetate, cis-hex-3-enyl acetate (mangosteen-like), furfuryl
methyl ketone, and cis-hex-3-en-1-ol. Compounds contributing significantly
to the aroma note of mangosteen were listed as hexyl acetate, cis-hex-3-enyl
acetate, and cis-hex-3-en-1-ol, respectively (MacLeod and Pieris, 1982).

17.2.7  Snake Fruit (Salacca edulis Reinw)

Snake fruit belongs to the class Salacca and originated from Southeast Asia.
The egglike fruit with a snakelike brown skin (Table 17.2) has a distinct
sour-sweet aroma note (Supriyadi et al., 2002). There are many cultivars

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Table 17.2  Some Exotic Tropical Fruits

Name Fruit Location

Yellow passion Cultivated in

(Passiflora Brazil,
edulis f. Hawaii, Fiji,
Flavicarpa) and Taiwan
(Marostica
and
Pastore,
2007)

Mangosteen Indian
(Garcinia subcontinent,
mangostana) Malaysia
(MacLeod
and Pieris,
1982)

Pitanga (Eugenia Brazil,

uniflora) Argentina,
Uruguay,
and
Paraguay
(Consolini
and
Sarubbio,
2002)

(Continued )

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Table 17.2  Some Exotic Tropical Fruits

Name Fruit Location

Mangaba Brazil
(Hancornia (Sampaio
speciosa and
Gomes) Nogueira,
2006)

Star fruit or Indonesia,

carambola Israel,
(Averrhoa Malaysia,
carambola L.) Central
America
(MacLeod
and Ames,
1990)

Snake fruit Southeast

(Salacca edulis Asia
Reinw) cv. (Supriyadi
Pondoh et al.,
2002)

(Continued )

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

Table 17.2  Some Exotic Tropical Fruits

Name Fruit Location

Camu-camu Brazil
(Myrciaria (Franco and
dubia) Shibamoto,
2000)

Cupuacu Brazil
(Theobroma (Franco and
grandiflorum) Shibamoto,
2000)

Araca-boi Brazil
(Eugenia (Franco and
stipitata) Shibamoto,
2000)

Guabiju South
(Myrcianthes America
pungens) (Marin
et al., 2008)

(Continued )

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

Table 17.2  Some Exotic Tropical Fruits

Name Fruit Location

Guabiroba South
(Campomanesia America
xanthocarpa) (Marin
et al.,
2008)

Lychee (Litchi China (Wu

chinensis) et al.,
2009)

African star Africa (Keay,

apple 1989)
(Chrysophyllum
albidum)

(Continued )

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

Table 17.2  Some Exotic Tropical Fruits

Name Fruit Location

Black velvet West and

tamarind Central
(Dialium Africa
guineense) (Achoba
et al.,
1992)

of snake fruit in Southeast Asia; however, most of them have an astringent

taste and are not sweet. The only sweet cultivar is the ‘Pondoh’. Analysis
of the volatile constituents of cv. ‘Pondoh’ with solvent-assisted flavor
evaporation (SAFE) and solvent extracts identified 109 volatile compounds
(Supriyadi et al., 2002). Some of these compounds comprised methyl esters
of butanoic acids, 2-methyl butanoic acid, hexanoic acids, pentanoic acid,
and the corresponding carboxylic acids. However, compounds contributing
significantly to the aroma of the snake fruit are methyl-3-methyl pentano-
ate, methyl-3-methyl-2-pentenoate, acetic acid, and 2-methyl butanoic acid.

17.2.8  Costa Rican Guava (Psidium friedrichsthalium)

Costa Rican guava is a shrub approximately 8 m high, found in Mexico and
Panama (Pino et al., 2002). The flavor of this guava has been described as
sweet with strong fruity, woody-spicy, and floral notes, with some green
and camphoraceous character (Pino et al., 2002). A gas chromatography–
mass spectrometry and olfactometry analysis of the fruit identified a total
of 173 volatile compounds (Pino et al., 2002). A quantitative analysis of
the volatile compounds revealed a large number of terpenes and terpenic
derivatives. The compounds contributing significantly to the aroma notes
are (E)-β-caryophyllene, α-terpineol, α-pinene, α-selinene, β-selinene,
δ-cadinene, 4,11-selinadiene, and α-copaene. Other compounds identified
are ethyl acetate, ethyl butanoate, ethyl hexanoate, butyl hexanoate, lin-
alool, (Z)-3-hexenol, and (Z)-3-hexenyl acetate. The following compounds

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

are thought to contribute to the complexity of the Costa Rican guava flavor.
The fruity notes are ascribed to the many aliphatic esters, whereas floral
note is attributed to α-terpineol and linalool. The woody and spicy note is
ascribed to the (E)-β-caryophyllene and its epoxide, whereas the green
note is attributed to (Z)-3-hexenol and (Z)-3-hexenyl acetate.

17.2.9  Pitanga Fruit (Eugenia uniflora L.)

The pitanga fruits are round, about 3 cm in diameter, with eight furrows
on the surface, and their color ranges from orange to purple (Table 17.2)
(Bezerra et al., 2000). Pitanga has an exotic and pleasant flavor. Apart from
the pleasant flavor, pitanga is used as an antidiarrheic, diuretic, and antirheu-
matic agent in Brazil (Schapoval et al., 1994). Analysis of the volatile constit-
uents of pitanga fruits identified 29 compounds comprising monoterpenes,
such as trans-β-ocimene, cis-ocimene, the isomeric β-ocimene, β-pinene
with woody-green pinelike odor, curzene, and bergaptene (Oliveira et al.,
2006). The major volatile compound of pitanga, trans-β-ocimene, is also an
important volatile constituent of the umbu-caja fruit (Spondias citherea).
Also, β-pinene is an important volatile constituent of both umbu-caja and
camu-camu (Myrciaria dubia) (Franco and Shibamoto, 2000).

17.2.10  Umbu-Caja Fruit (Spondias citherea)

Umbu-caja is a small round fruit (approximately 3 cm in diameter) with a
yellow skin and a refreshing aroma and sour flavor (Marin et al., 2008).
This fruit is native to the northeastern region of Brazil. High-resolution gas
chromatography–mass spectrometry analysis of the fruit identified 26 com-
pounds comprising primarily terpenes and their derivatives. Compounds
contributing significantly to the aroma of the fruit are cis-β-ocimene,
β-caryophyllene, trans-ocimene, δ-limonene, and α-caryophyllene (Franco
and Shibamoto, 2000). Also, three other volatiles, 1,3-cyclohexadien-1,5,5,6-
tetramethyl-1,3-cyclo-hexadiene, β-selinene, and 1,2,3,4,4a,5,6,8a-octa-
hydro-4a,8-dimethyl naphthalene (Table 17.1), present a distinctive flavor.

17.2.11  Camu-Camu Fruit (Myrciaria dubia)

This is a round fruit (2–2.5 cm in diameter) with a red skin and pink pulp
(Franco and Shibamoto, 2000). Camu-camu has a distinctive acidic flavor.
Isolation and characterization of odor-active compounds in camu-camu by
gas chromatography–olfactory (GC-O) and high-resolution gas chroma-
tography–mass spectrometry (HRGC-MS) identified 20 aroma compounds

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

(Franco and Shibamoto, 2000). The majority of the volatile compounds

identified in camu-camu are terpenes, such as α-pinene, δ-limonene,
β-caryophyllene, α-fenchene, car-3-ene, y-terpine and p-cimene. Other
compounds contributing to the aroma of camu-camu are the monoterpene
alcohols, such as eucalyptol, fenchol, and terpineol.

17.2.12  Cupuacu Fruit (Theobroma grandiflorum)

Cupuacu fruits are 12–25 cm in length and 10–12 cm in diameter. It is an
oblong fruit with a hard skin and creamy white pulp (Franco and Shibamoto,
2000). Cupuacu is an Amazonian forest fruit from Brazil. HRGC-MS anal-
ysis of the volatile compounds of the fruit identified 21 compounds. The
compounds contributing to the aroma note of the fruit are ethyl butano-
ate and ethyl hexanoate (Franco and Shibamoto, 2000). Other researchers
(Alves and Jennings, 1978; Fischer et al., 1993) have earlier identified ethyl
butyrate, ethyl-2-methyl butyrate, 1-butanol, ethyl hexanoate, 3-hydroxyl-
2-butanone, ethyl octanoate, acetic acid, and linalool in cupuacu fruits with
the aid of the Solid phase extraction (SPE) technique (Fischer et al., 1993).

17.2.13  Araca-Boi Fruit (Eugenia stipitata)

Araca-boi is a yellow round fruit, about 12 cm in diameter (Franco and
Shibamoto, 2000). The fruit has a distinctive acidic flavor. Analysis of the
volatile constituents of the fruit with high-resolution gas chromatography–­
mass spectrometry (HRGC-MS) identified 31 compounds compris-
ing a complex pattern of sesquiterpenes (Franco and Shibamoto, 2000).
Compounds contributing significantly to the aroma note of araca-boi are
germacrene D, β-pinene, δ-cadinene, and α-pinene, respectively.

17.2.14  Mangaba Fruit (Hancornia speciosa Gomes)

Mangaba is a Brazilian tropical fruit with an exotic flavor and aroma. The
fruit is an ellipsoid or spherical berry, 2.5–6 cm in length, with white, sweet
pulp (Vieira-Neto, 2002). A total of 32 volatile compounds were identified
in mangaba fruit (Sampaio and Nogueira, 2006). The compounds comprise
3-hydroxy-2-butanone, 2,4,5-trimethyl-1,3-dioxolane, 3-methyl-3-buten-
1-ol, 3-methyl-1-butanol, furfural, 3-methyl-1-butanyl acetate, 3-methyl-
3-buten-1-yl acetate, (Z)-linalool oxide, (E)-linalool oxide, linalool, 2-phenyl
ethanol, α-terpineol, geraniol, octadecanol, 1-octen-3-ol, and hexadecanal.
The compounds that contribute markedly to the fruity note of mangaba are
the esters, alcohols, aldehydes, and ketones (Mathesis et al., 1992).

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17.2.15  Garcinia Fruit (Garcinia dulcis Kurz)

Garcinia or rata fruit belongs to the family Guttiferae and is probably native
to the Philippines or Borneo. The fruit is about 2.5 cm in diameter, with
a short, sharp end (Pino et al., 2003). The soft flesh of the ripe fruit has
a butterlike consistency and a pleasant acidic flavor. Analysis of the vola-
tile constituents identified 90 volatile compounds made up of terpenoids,
alcohols, esters, and furanoids (Pino et al., 2003). Odor description of the
fruit varies from floral-woody to acidic. The compounds contributing to this
odor note included linalool, geraniol, α-terpineol, (E)-linalool oxide, ter-
pinolene, (Z)-linalool oxide, (E)-β-ocimene, β-pinene, hexanoic acid, and
2-methyl-3-buten-2-ol.

17.2.16  Guabiju Fruit (Myrcianthes pungens Berg)

Guabiju fruit belongs to the Myrtaceous family and is native to Brazil
(Marin et al., 2008). Other members of this family are pitanga (Eugenia
uniflora), araca (Psidium cattleyanum), guabiroba (Campomanesia
­xanthocarpa) and cereja-do-rio-grande (Eugenia involucrate). Guabiju is
oblong in shape and has a purple color when ripe. Analysis of the vola-
tile constituents of guabiju identified primarily sesquiterpenoids, such
as β-caryophyllene, germacrene D, bicyclogermacrene, α-humulene, and
γ-cadinene (Marin et al., 2008). Other important contributors to the aroma
note are the oxygenated sesquiterpenes, such as β-eudesmol, epi-globulol,
and elemol.

17.2.17  Guabiroba Fruit (Campomanesia xanthocarpa Berg)

Guabiroba fruit is also a member of the Myrtaceous family. This is a fruit
with a smooth dark yellow epidermis that resembles guava (Marin et al.,
2008). It is native to South America. Analysis of the volatile constituents of
guabiroba fruit identified 38 compounds, which included β-caryophyllene,
α-humulene, bicyclogermacrene, limonene, α-pinene, β-pinene, guaiol, and
hexadecanoic acid as the main contributors to the fruit aroma notes.

17.2.18  Bacuri Fruit (Platonia sculenta)

Bacuri fruits are ovoid to subglobose and weigh between 200 and 1000 g
(Marostica and Pastore, 2007). The fruit has a creamy white mucilaginous

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

pulp with a very attractive exotic flavor. Analysis of the volatile constitu-
ents of the fruit revealed linalool, 2-heptanone, and cis-3-hexenyl acetate
as the most potent flavor compounds (Alves and Jennings, 1979). However,
analysis of the free and bound flavor components of bacuri fruit with GC
and GC-MS using XAD-amberlite separation identified 75 compounds
(Boulanger et al., 1999). The compounds contributing significantly to the
aroma are the terpene alcohols, such as α-terpineol, hotrienol, nerol oxide,
nerol, geraniol, and linalool.

17.2.19  Cashew Fruit (Anacardium occidentale L.)

Cashew is an indigenous tree of tropical America with a strong indication
that its center of origin is the coastal strip of north and northeast Brazil,
where its fruits are called caju (Maia et al., 2000). Some examples of
impact-flavor compounds have already been identified in cashew fruits.
These are δ-carene, limonene, trans-hex-2-enal (MacLeod and Troconis,
1982), furfural, and 4-hydroxydodecanoic acid lactones (Maia et al., 2000).
Essences of yellow and red cashew fruits obtained by simultaneous distilla-
tion–extraction using a Chrompack Micro-Steam distillation extractor and
pentane revealed that the total amount of C 6 aldehydes, alcohols, and acids
comprised 26% and 30% of the essence of fresh yellow and red cashew fruits,
respectively (Maia et al., 2000). An earlier report on the volatile constitu-
ents of the Venezuela cashew identified δ-carene, limonene, trans-hex-2-
enal, hexenal, and furfural as the main aroma components (MacLeod and
Troconis, 1982).

17.2.20  Melon Fruit (Cucumis melo)

Melon comprises a great number of varieties, and they exhibited a wide
variation in flavor and aroma profiles (Jordan et al., 2001). Currently, seven
well-known cultivars of Cucumis melo are cultivated in the United States,
but members of the Reticulatus (var. reticulates Ser) are probably the most
commercially important melons. The characteristic aroma components
of melon were previously identified as methyl (methylthio) acetate, ethyl
(methylthio) acetate, 3-(methylthio) propanitrile, 3-(methylthio) propanol,
and 2-(methylthio) ethanol acetate (Wyllie and Leach, 1992). The aroma
volatiles of some melon species consist of a complex mixture of esters
together with other components, including C 9 unsaturated aldehydes, alco-
hols, and acetates, whose sensory properties have been described as mel-
onlike (Homatidou et al., 1992; Wyllie and Leach, 1992; Wyllie et al., 1994).
More recently, a comparative study of the aromatic profile of muskmelon

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

aqueous essence and the puree of fresh fruit using gas chromatography–
mass spectrometry (GC-MS) and gas chromatography–olfactometry
(GC-O) revealed 53 components in the essence and 38 in the fresh fruit,
respectively (Jordan et al., 2001). The compounds contributing significantly
to the melon aroma included 2-methyl-3-buten-2-ol, 2,3-butanediol, methyl-
3-phenyl propionate, ethyl-3-phenyl propionate, and ethyl-3-(methylthio)
propionate.

17.2.21 Jackfruit (Artocarpus heterophyllus Lam.)

Jackfruit is assumed to have its origin in India or Oceania, and it is found as
a cultivated tree in all tropical countries (Rasmussen, 1983). Jackfruit can
be eaten raw, salted as pickle, cooked, or sweet (Maia et al., 2004). There are
two known varieties of jackfruit. These are the hard and soft jackfruits. The
aroma of jackfruit is very distinguishable. A study of the aroma components
of two varieties of jackfruit using gas chromatography–mass spectrometry
identified isopentyl isovalerate, butyl isovalerate, palmitic acid, and ethyl
isovalerate as the major components in the hard variety. The aroma profile
of the soft variety was dominated by isopentyl isovalerate, butyl acetate,
butyl isovalerate, and isopentan-1-ol (Maia et al., 2004).

17.2.22  Sapodilla Fruit (Achras sapota L.)

This is an edible fruit with a characteristic and most attractive delicate fla-
vor. The fruit is very sweet and best consumed raw, but fully ripe. When
underripe, it has a rather unappealing alcoholic aftertaste. Although per-
haps not well-known elsewhere, sapodilla is considered one of the best
fruits of the American tropics. It is grown on a commercial scale in some
Central American countries, but produce is marketed fresh, and currently
the fruit is not processed in any way, although the juice provides a most
refreshing drink, which might have commercial potential. A recent study
on the volatile constituents of sapodilla fruit, using headspace solid-phase
microextraction (HS-SPME) with the gas chromatography–mass spec-
trometry (GC-MS) technique, revealed 23 volatile compounds (Laohakunjit
et al., 2006). Compounds contributing significantly to the aroma note
included ethyl acetate, acetaldehyde, benzyl alcohol, and 2-butenylben-
zene. However, these results were inconsistent with the study of MacLeod
and Troconis (1982), who reported methyl benzoate and methyl salicylate
as the major contributors to the sapodilla fruit aroma. Differences in variet-
ies might have contributed to the results reported by the different groups
of researchers.

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

17.2.23  Genipap Fruit (Genipa americana)

Genipap is an elliptical fruit, about 7–9 cm in diameter, with a pale brown-
ish color. Because of its extremely strong flavor, it is consumed mainly
as liquor, jelly, juice, and ice cream (Pinto et al., 2006). A bibliographical
search indicated that aroma composition of tropical genipap has been the
subject of only two indexed previous publications (Borges and Resende,
2000; Pinto et al., 2006). Borges and Resende (2000) identified 35 vola-
tiles from fruits obtained in Brazilian Amazonia. The application of the
gas ­chromatography–olfactrometry and aroma extract dilution analysis
(AEDA) techniques to the fruit extracts allowed identification of butyric,
2-methyl butyric, and hexanoic acids as the odorants responsible for the
harsh and acidic notes. The typical fruity note perceived in genipap aroma
was attributed to ethyl-2- and ethyl-3-methyl butyrate. These results were
based solely on analyses of genipap aroma extracts obtained by a simultane-
ous steam distillation and dichloromethane extraction method (Borges and
Resende, 2000). Recent analysis of the volatile constituents of genipap fruit
extracts by both headspace and liquid-phase adsorptive chromatography
and gas chromatography–flame ionization detection (GC-FID), GC–mass
spectrometry (GC-MS), and GC–olfactrometry–AEDA revealed 52 vola-
tiles. Among these are hexanoic and 2- and 3-methyl butanoic acids and the
corresponding methyl butanoic esters, methyl hexanoate, methyl and ethyl
octanoate, acetic acid, and 2-methyl propanoic acids (Pinto et al., 2006).

17.2.24  Soursop Fruit (Annona muricata)

Soursop is a tropical fruit native to and common in tropical America and the
West Indies (MacLeod and Pieris, 1981). Soursop is cherished for its pleasant
and distinctive aroma, described as a custardlike flavor (McGorrin, 2007).
The fruit has been widely used for manufacturing various kinds of products,
such as puree, syrups, jams, and ice cream (Umme et al., 1999). A recent
study on the equilibrium headspace analysis of volatile flavor compounds
extracted from soursop using solid-phase microextraction (Cheong et al.,
2010) identified 35 compounds, comprising primarily esters. The esters
were methyl butanoate, methyl hexanoate, methyl-(E)-2-butenoate, methyl-
(E)-2-hexenoate, hydroxyl methyl esters, ­ methyl-3-hydroxybutanoate,
methyl-3-hydroxyhexanoate, methyl-2-hydroxy-3-methyl butanoate, and
methyl-2-hydroxy-3-methyl pentanoate. Other compounds contributing to
the aroma note included limonene, linalool, caryophyllene, and methyl-(E)-
cinnamate. In a previous study (MacLeod and Pieris, 1981) on the essence
of soursop using a low-temperature high-vacuum distillation procedure
and gas chromatography–mass spectrometry, results revealed that methyl

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P OST H AR V EST RI P ENING P H Y SIOLOG Y

hexanoate and methyl hex-2-enoate were the two most abundant compo-
nents of soursop.

17.2.25  Acerola Fruit (Malphigia glabra L.)

Acerola fruit is known under various names, for example, West Indian
cherry and Barbados cherry (Gomez et al., 1999). It is a small trilobite red
fruit and quite similar to a cherry. The pulp is very juicy and cooling and
possesses a fruity and sweet flavor. A bibliographical search indicated that
aroma composition of acerola has been the subject of only three indexed
previous publications (Schippa et al., 1993; Boulanger and Crouzet, 2001;
Carasek and Pawliszyn, 2006). Boulanger and Crouzet (2001) identified
46 volatile compounds in free and glycosidically bound flavor compounds
of acerola. Alcohols (3-methyl-but-3-en-1-ol, 3-methyl-butan-1-ol, and
2-methyl-butan-1-ol) were predominant in the volatile fraction. Other stud-
ies (Schippa et al., 1993; Carasek and Pawliszyn, 2006) similarly identified
these alcohols in acerola fruit extracts. In addition to these alcohols, six
derived esters of these alcohols were identified as major contributors to the
aroma note of acerola fruit. Boulanger and Crouzet (2001) also identified
42  aglycones in the fruit. Among these aglycones are the norisoprenoids
and aliphatic alcohols. Hydrolysis of these aglycones could increase the
fruity aroma of acerola.

17.2.26  Tamarind Fruit (Tamarindus indica L.)

Tamarind is native to tropical Africa, but distributed throughout the tropi-
cal countries of the world. The fruit is flat and beanlike in shape. Tamarind
is high in sugar and minerals, with a pleasant acid taste and aroma (Carasek
and Pawliszyn, 2006). A study of the volatile constituents of tamarind using
automated headspace solid-phase microextraction (HS-SPME) coupled
with gas chromatography–mass spectrometry (GC-MS) identified 27 vola-
tile compounds. Aldehydes and esters were the predominant volatiles in the
tamarind. The most significant volatile compounds included phenyl acet-
aldehyde, furfural, (Z)-3-hexenyl acetate, hexyl acetate, geranyl acetone,
limonene, and caryophyllene (Carasek and Pawliszyn, 2006).

17.2.27  Velvet Tamarind (Dialium guineense)

Velvet tamarind is a fruit tree commonly found in West and Central Africa.
It is called Awin by the Yoruba in the southwestern part of Nigeria, whereas
in the southeastern part, it is called Icheku or Nchichi. It is a woody plant

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belonging to the family Ceasalpiniaceae. The fruit is characterized by a

velvety skin and a brown or black color. The seeds are housed inside a dry,
sweet, but slightly astringent orange-red pulp that is eaten raw. The fruit
are harvested during the dry season and often serve as a form of thirst
quencher for the locals. Chemical analysis of the pulp reports a low mois-
ture and protein content, but a high carbohydrate content. Also, it is rich
in certain minerals, such as ascorbic acid and α-carotene. Presently the
fruit is still highly underutilized because of its short harvest season, which
peaks around March and April. A recent study of the aroma components
of two varieties of velvet tamarind using gas chromatography–mass spec-
trometry identified 2-methoxyethyl cyanoacetate, pentadecanal, 4-acetyl-
pyrazole, furfural, and 5-methyl furfural as the predominant volatiles in the
fruit (Lasekan, unpublished).

17.2.28  African Star Apple Fruit (Chrysophillum albidum)

African star apple is a large berry containing four to five flattened seeds
(Keay, 1989). The fruit belongs to the family Sapotaceae. HS-SPME-GC-MS
analyses of the juices obtained from four different varieties of African star
apples (Chrysophyllum albidum) led to the identification of 59 odor-active
compounds. However, the application of the aroma extract dilution analy-
sis (AEDA) on the fresh juice revealed 45 odor-active compounds in the
Flavour dilution (FD) factor range of 4–128. A great variety of odor quali-
ties, such as fruity, grassy, woody, and citruslike, were perceived. Sniffing
of serial dilutions of the extract revealed the highest FD factors for the pine-
apple, apricot-like methyl hexanoate; cherrylike acetophenone; and ethyl
dodecanoate, which elicit a sweet floral nuance. Lower FD factors were
obtained for the fruity, ethyl acetate, ethyl-(E)-2-butenoate, ethyl hexano-
ate, methyl benzoate, and hexyl butanoate (Lasekan et al., 2013).

17.3  Aroma Compounds’ Biosynthetic

Pathways and Metabolism

17.3.1 Fruit’s Volatile Ester Metabolism

Volatile esters are produced by virtually all soft fruit species during ripen-
ing. In some fruits, such as rambutan (Nephelium lappaceum L.), carambola
(Averrhoa carambola L.), jackfruit (Artocarpus heterophyllus Lam.), apple
(Malus domestica), pear (Pyrus communis), and banana (Musa sapientum),
esters are the major components in their characteristic aroma. In other
fruits, such as strawberry (Fragaria × ananassa), they contribute to the

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blend of volatiles that constitute the aroma. Often, a single fruit emits a
large spectrum of esters; for instance, more than 100 different esters have
been detected in ripe strawberry fruit (Zabetakis and Holden, 1997). Most
volatile esters have flavor characteristics described as fruity (Burdock,
2005). However, they not only are produced and emitted by fruit but also
are part of floral scents (Dudareva et al., 1996). Formation of volatile esters
is not restricted to the plant kingdom and is also common in microorgan-
isms such as yeast and fungi.
The metabolic pathways for volatile biosynthesis in fruits are not fully
understood. More than 80% of the volatiles produced by ripe strawberry
and melon are esters (Dirinck et al., 1989). Therefore, production of these
compounds has been the major focus of research in the past. Despite the
diversity of volatile compounds among these fruits, the basic biosynthetic
pathways may be similar. The metabolism of fatty acids and branched
amino acids may serve as a precursor for the biosynthesis of aroma vola-
tiles in fruits (Perez et al., 2002; Fellman et al., 2003). Fatty acids play a
major role in ester synthesis. According to Rowan et al. (1999), straight-
chain ester volatiles in whole fruit arise predominantly by β-oxidation of
fatty acids. It is widely assumed that lipoxygenase (LOX) may contribute
to the breakdown of long-chain fatty acids to C 6 aldehydes, which are con-
verted to alcohols by aldehyde dehydrogenase (Rowan et al., 1999; Fellman
et al., 2003). The alcohols are subsequently converted to hexyl and hexenyl
esters (Shalit et al., 2001; Perez et al., 2002).
Such a pathway has been demonstrated by Wang et al. (1996), who
altered the levels of fatty acids in transgenic tomato (Lycopersicon ­esculentum)
fruit, leading to dramatic changes in the aroma profile. Ripening-related
processes, such as changes in cell wall composition, might also contribute
intermediates to ester biogenesis. In tomato, evidence was provided that
the activity of pectin methyl esterase (which catalyzes demetoxylation of
pectin) regulates levels of methanol in the fruit (Frankel et al., 1998). The
accumulating methanol may then be utilized to generate methyl esters in
ripe fruit.
Branched amino acids are important substrates for branched-chain
volatiles such as 2-methyl butyl acetate and ethyl-2-methyl butanoate. This
pathway was shown to be present in both apple and strawberry (Perez et al.,
2002). For example, in strawberry, the amino acid alanine has been impli-
cated in the formation of ethyl esters during ripening (Perez et al., 1992).
However, the levels of amino acid precursors could not explain, in all cases,
the formation of the corresponding ester, and it has therefore been sug-
gested that the selectivity of enzymes preceding alcohol acyl transferase
(AAT) in the biosynthetic pathway also plays a role in determining ester
composition (Song and Forney, 2008). The enzyme that is responsible
for the final step of ester formation is AAT, which combines alcohols and

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 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

acyl-CoAs to form esters. Combinations between different alcohols and

acyl-CoAs will result in the formation of a range of esters in different fruit
species. Other enzymes, such as LOX, alcohol dehydrogenase (ADH), and
pyruvate decarboxylase (PDC), are believed to be involved in the pathways
to provide aldehydes and alcohols for ester synthesis (Fellman et al., 2000).

17.3.2  Fruit’s Volatile Terpenoid Metabolism

Volatile terpenoids represented by mainly isoprene (C 5), monoterpenes
(C10), and sesquiterpenes (C15) constitute the largest class of plant vola-
tile compounds. They play important roles in direct and indirect plant
defense against herbivores and pathogens. Also, they play important roles
by attracting pollinators and seed disseminators in plant thermotolerance
(Dudareva et al., 2006). Apart from their importance in plant physiology
and ecology, volatile terpenoids are also used as natural flavor and aroma
compounds and have a beneficial impact on humans as health-promoting
compounds (Wagner and Elmadfa, 2003). All terpenoids are synthesized
from the universal 5-carbon precursors, isopentenyl diphosphate (IPP) and
dimethyl allyl diphosphate (DMAPP), which are derived from two alterna-
tive biosynthetic pathways localized in different subcellular compartments
(Figure 17.1). While the DMAPP formed in plastids is used by isoprene syn-
thase (ISPS) to form isoprene in some plants, IPP and DMAPP precursors
are further condensed by prenyl diphosphate synthases in the respective
compartments to form prenyl diphosphate intermediates serving as sub-
strates for a large group of terpene synthase (TPS) enzymes, resulting in
the final terpenoid compounds (Dudareva et al., 2006).
In plants, two biosynthetic pathways are responsible for the synthesis
of IPP and DMAPP, the universal precursors of all terpenoids. The classical
cytosolic mevalonic acid (MVA) pathway gives rise to IPP from acetyl-CoA
(Newman and Chappell, 1999), whereas the plastidial 2-C-methylerythritol-
4-phosphate (MEP) pathway (Rohmer et al., 1993) described during the
1990s leads to the formation of IPP and DMAPP from pyruvate and glycer-
aldehyde-3-phosphate (Figure 17.1). Although the subcellular compartmen-
talization of two pathways allows them to operate independently, metabolic
cross talk between the two pathways has been reported (Hemmerlin et al.,
2003). Following the formation of IPP and DMAPP, prenyl transferases
located in both compartments utilize IPP and DMAPP to produce prenyl
diphosphates (Figure 17.1). Condensation of one molecule of DMAPP and
two molecules of IPP catalyzed by farnesyl diphosphate synthase (FDS)
leads to the formation of farnesyl diphosphate (FPP) (C15), the natural
precursor of sesquiterpenes (Dudareva et al., 2006). Meanwhile, geranyl
diphosphate synthase (GDS) catalyzes the condensation of a molecule of

575
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Cytosol / ER / Peroxisomes
Pyruvate
Acetyl-CoA +GA-3P
AACT DXS
AcAc-CoA DOXP
HMGS DXR

HMG-CoA MEP
MEP MCT
MVA HMGR PATHWAY
CDP-ME
PATHWAY Mevalonate (MVA)
CMK
MVK
CDP-ME2P Isoprene
Mevalonate-5-phosphate Plastid
MDS
PMK ISPS
ME-2,4cPP
Mevalonate-5-diphosphate
HDS
MVD HMBPP
IDI HDR HDR
IDI
DMAPP IPP IPP DMAPP
IDI
GGDS FDS
GDS IPP DMAPP GDS
FDS
GGPP
ria

GGDS FDS FPP GPP

GPP FPP
nd

TPS TPS TPS

TPS
cho

GGPP FPP Mono

TPS
to

Mono terpenes
Mi

Sesqui Sesqui
terpenes Sesquiterpenes terpenes
terpenes

Figure 17.1  Biosynthetic pathways and their compartmentalization leading

to volatile terpenoid formation in plants. AACT, acetoacetyl-CoA thiolase;
AcAc-CoA, acetoacetyl-CoA; CDP-ME, 4-(cytidine-50-diphospho)-2-C-
methyl-D-erythritol; CDP-ME2P, 4-(cytidine-50-diphospho)-2-C-methyl-
D-erythritol phosphate; CMK, CDP-ME kinase; DMAPP, dimethyl allyl
diphosphate; DOXP, 1-deoxy-D-xylulose-5-phosphate; DXR, DOXP reductoi-
somerase; DXS, DOXP synthase; FDS, farnesyl diphosphate synthase; FPP,
farnesyl diphosphate; GA-3P, glyceraldehyde-3-phosphate; GDS, gera-
nyl diphosphate synthase; GGDS, geranyl geranyl diphosphate synthase;
GGPP, geranyl geranyl diphosphate; GPP, geranyl diphosphate; HDR, (E)-4-
hydroxy-3-methyl but-2-enyl diphosphate reductase; HDS, (E)-4-hydroxy-3-
methyl but-2-enyl diphosphate synthase; HMBPP, (E)-4-hydroxy-3-methyl
but-2-enyl diphosphate; HMG-CoA, 3-hydroxy-3-methylglutaryl-CoA;
HMGR, HMG-CoA reductase; HMGS, HMG-CoA synthase; IDI, isopen-
tenyl diphosphate isomerase; IPP, isopentenyl diphosphate; ISPS, isoprene
synthase; MCT, 2-C-methyl-D-erythritol-4-phosphate cytidylyl transferase;
MDS, 2-C-methyl-D-erythritol-2,4-cyclodiphosphate synthase; ME-2,4cPP,
2-C-methyl-D-erythritol-2,4-cyclodiphosphate; MEP, 2-C-methyl-D-erythritol-
4-phosphate; MVD, mevalonate diphosphate decarboxylase; MVK, meva-
lonate kinase; PMK, phosphomevalonate kinase; TPS, terpene synthase.
Names of the enzymes are boxed and volatile terpenoids are underlined.
(From Nagegowda, D.A., FEBS Letters 584, 2965–2973, 2010.)

576
 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

IPP and DMAPP to produce geranyl diphosphate (GPP) (C10), the precur-
sor of monoterpenes.

17.3.3  Sulfur Volatile Compounds’ Biosynthetic Pathway

Volatile sulfur compounds (VSCs) have an unpleasant sulfurous odor at
higher levels, and therefore have a highly dependent olfactive profile. These
compounds also have a very low flavor threshold value of 0.1 part per tril-
lion (ppt) in water and are generally considered key odorants (Iranshahi,
2012). Because of this, VSCs play an important role in determining the fla-
vor and odor properties of many foodstuffs, as well as providing the odor
of some oils, such as rose oil. The chemical structures of VSCs vary from
the simple, such as that seen in methanthiol and dimethyl sulfide, to the
more complicated compounds, such as that of 4,5-­epithiocaryophyllene.
The more complicated structures occasionally contain other hetero-
atoms, such as nitrogen. This structural motif is seen in β-mint sulfide,
4,5-­epithiocaryophyllene, and 5-(4-acetoxy-1-butynyl)-2,2′-bithiophene.
The biosynthetic pathways of a large number of VSCs found in terres-
trial plants have not yet been examined. To date, most investigations of VSC
biosynthetic pathways have concentrated on the thiosulfinates of Allium
species and the isothiocyanates of the Brassicaceae family. Although many
VSCs have been identified, many of the enzymes and genes involved in their
biosynthesis are still unknown (Schwab et al., 2008). The characteristic fla-
vor of Allium species is due to their VSCs, which are produced by enzymatic
decomposition of S-alk(en)yl-L-cysteine-S-oxides, such as methiin, propiin,
alliin, and isoalliin, which are present as flavor precursors (Figure 17.2)
(Fritsch and Keusgen, 2006). When fresh garlic (or other Allium species) is
chopped or crushed, the enzyme alliinase converts alliin into allicin, which
is primarily responsible for the aroma of fresh garlic. Some other cysteine
sulfoxides, such as ethiin and butiin, have also been reported from Allium
species. In fact, in onion and garlic, series of VSCs are formed by cleav-
age of odorless S-alk(en)yl cysteine sulfoxide flavor precursors catalyzed
by the enzymes alliinase and lachrymatory-factor synthase. VSCs originat-
ing from methionine and cysteine are responsible for the odor of dimethyl
disulfide, cooked cabbage (methanethiol), and boiled potatoes (methional).
The biosynthesis of glucosinolates is started by the N-hydroxylation of a
precursor amino acid (α-amino acid), followed by decarboxylation to form
an aldoxime. The aldoxime intermediate then rearranges to form thiohy-
droxamic acid by an unknown mechanism. Biosynthetic steps after thio-
hydroxamic acid formation include an introduction of the thioglucoside
sulfur from cysteine, S-glycosyl transfer from UDP-glucose, and sulfina-
tion by 3′-phosphoadenosine-5′-phosphosulfate, a well-known sulfate donor
(Figure 17.2). In fact, isothiocyanates, thiocyanates, and nitriles are the

577
P OST H AR V EST RI P ENING P H Y SIOLOG Y

H2 O
O NH2 O
alliinase S
S R OH S R Thiosulfinates
R COOH R S
S-alk(en)yl cysteine sulfoxides Sulfenic acid
(e.g. methiin, ethiin, butiin, alliin intermediates
isoalliin, propiin)
S O O
S
S R
R S
R
S R S R
S S R S
S R S R

COOH SH– UDP-Glc

SH
R NH2 OH
R N
R NOH
α-amino acid Aldoxime
CH2OH
O
OH
Myrosinase
N S R S OH
R C OH
Isothiocyanates N
OSO3–
Glucosinolate
S N
Thiocyanates R C

S
C
Epithionitriles N

Figure 17.2  Biosynthetic pathways for thiosulfinates and isothiocyanates

(From Iranshahi, M., Journal of Essential Oil Research 24, 393–434, 2012.)

products of the enzymatic breakdown of the related glucosinolates. The

enzyme cleavage process has been shown to be catalyzed by myrosinase
(Fahey et al., 2001). The sulfate moiety is then released in a nonenzymatic
fashion from glucosinolates. Methylthio-, methyl sulfinyl- and methyl sul-
fonyl-isothiocyanates form a large family of isothiocyanates. Conversion
of methylthio-glucosinolates to their methyl sulfinyl and methyl sulfonyl
homologues is carried out by an oxidation process. Like other isothiocya-
nates, methylthio-, methyl sulfinyl-, and methyl sulfonyl-isothiocyanates
are formed from the breakdown of the related glucosinolates. However, the
exact mechanism of the side-chain modification of glucosinolates and iso-
thiocyanates has not been fully understood (Fahey et al., 2001).

578
 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

17.4  Conclusions and Future Perspectives

Exotic tropical fruits can be conveniently classified into two broad groups on
the basis of whether esters or terpenes predominate in the aroma. The cur-
rent review has revealed that the majority of the fruits belongs to the latter
group, in which terpenes predominate. For instance, most of the Amazonian
fruits, such as pitanga, umbu-caja, camu-camu, araca-boi, garcinia, guabiju,
guabiroba, and bacuri, belong to the terpene group. Other members of the
terpene group are the lychee and mangosteen. The ester group is made up
of rambutan, durians, star fruit, snake fruit, cupuacu, acerola, tamarind,
soursop, sapodilla, genipap, cashew, melon, jackfruit, and yellow passion
fruit. While yellow passion fruit is grouped with the ester-producing fruits,
it is worth noting that its volatile composition is rather complex and attrib-
uted to the presence of trace levels of sulfur volatiles in combination with
other compounds. Also, summaries of recent progress in the characteriza-
tion of esters, terpenoids, and sulfur volatiles’ biosynthetic genes and their
spatiotemporal expression patterns were reviewed.

References
Achoba, I.I., Ilori, J.A., Elegbede, J.A., and Kagbu, J.A. 1992. Nutrient com-
position of black velvet-tamarind seed and pulp from Nigeria. Journal of
Food Biochemistry 16, 229–233.
Alves, S., and Jennings, W.C. 1979. Volatile composition of certain Amazonian
fruits. Food Chemistry 4, 149–159.
Bezerra, J.E.F., Junior, J.F.S., and Lederman, I.E. 2000. Botanical and ecology. In
Pitanga (Eugenia uniflora), Donadio, L.C. (ed.). Jabotical, Brazil, 30–50.
Boulanger, R., and Crouzet, J. 2001. Identification of the aroma components
of acerola (Malphigia glabra L.): free and bound flavour compounds.
Food Chemistry 74, 209–216.
Boulanger, R., Chassagne, D., and Crouzet, J. 1999. Volatile compounds of the
free and bound bacuri fruits. Flavour and Fragrance Journal 14, 303–305.
Borges, E.C., and Resende, C.M. 2000. Main aroma constituents of genipap
(Genipa americana) and bacuri (Platonia sculenta). Journal of Essential
Oil Research 12, 71–74.
Burdock, G.A. 2005. Fenaroli’s Handbook of Flavour Ingredients. 5th ed. CRC
Press, Boca Raton, FL.
Buttery, R.G. 1981. Vegetables and fruit flavours. In Flavour Research: Recent
Advances, Teranishi, R., Flath, R.A., and Sugisawa, H. (eds.). Dekker,
New York, 175–216.
Carasek, E., and Pawliszyn, J. 2006. Screening of tropical fruit volatile compounds
using solid-phase microextraction (SPME) fibres and internally cooled
SPME fibre. Journal of Agricultural and Food Chemistry 54, 8688–8696.

579
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Cheong, K.W., Tan, C.P., Mirhosseini, H., Hamid, N.S., Osman, A., and Basri,
M. 2010. Equilibrium headspace analysis of volatile flavour compounds
extracted from soursop (Annona muricata) using solid-phase microex-
traction. Food Research International 43, 1267–1276.
Chin, S.T., Nazimah, S.A.H., Quek, S.Y., Che Man, Y., Rahman, R.A., and Mat
Hashim, D. 2008. Changes of volatiles attribute in durian pulp during
freeze- and spray-drying process. LWT Food Science and Technology 41,
1899–1905.
Consolini, A.E., and Sarubbio, M.G. 2002. Pharmacological effects of Eugenia
uniflora (Myrtaceae) aqueous crude extract on rat heart. Journal of
Ethnopharmacology 81(1), 57–63.
Dirinck, P., De Pooter, H., and Schamp, N. 1989. Aroma development in rip-
ening fruits. In Flavour Chemistry: Trends and Development, Teranishi,
R., Buttery, R., and Shahidi, F. (eds.). American Chemical Society,
Washington, DC, 24–34.
Dudareva, N., Cseke, L., Blanc, V.M., and Pichersky, E. 1996. Evolution of flo-
ral scent in Clarkia: Novel patterns of S-linalool synthase gene expres-
sion in the C. breweri flower. Plant Cell 8, 1137–1148.
Dudareva, N., Negre, F., Nagegowda, D.A., and Orlova, I. 2006. Plant vola-
tiles: recent advances and future perspectives. Critical Reviews in Plant
Science 25, 417–440.
Enzell, C.R. 1981. Volatile constituents of β-carotene degradation. In Flavour,
Schreier, P. (ed.). Walter de Gruyter, Berlin, 449–453.
Fahey, J.W., Zalcmann, A.T., and Talalay, P. 2001. The chemical diversity
and distribution of glucosinolates and isothiocyanates among plants.
Phytochemistry 56, 5–51.
Fellman, J.K., Miller, T.W., Mattinson, D.S., and Mattheis, J.P. 2000. Factors
that influence biosynthesis of volatile flavour compound in apple fruits.
Journal of Horticulture Science 35, 1026–1033.
Fellman, J.K., Rudell, D.R., Mattinson, D.S., and Matthesis, J.P. 2003.
Relationship of harvest maturity to flavour regeneration after CA stor-
age of ‘Delicious’ apples. Postharvest Biology and Technology 27, 39–51.
Fischer, N., Hammerschmidt, F.J., and Brunke, E.J. 1993. Volatile constitu-
ents of cupuacu (Theobroma grandiflorum). In Progression Flavour
Precursors Studies, Schreier, P., and Winter Halter, P. (eds.). Allured
Publishing, Coral Stream, IL, 287–301.
Franco, M.R.B., and Shibamoto, T. 2000. Volatile composition of some
Brazilian fruits: umbu-caja (Spondias cithera), camu-camu (Myrciaria
dubia), araca-boi (Eugenia stipitata) and cupuacu (Theobroma grandi­
florum). Journal of Agricultural and Food Chemistry 48, 1263–1265.
Frankel, E.N., Bosanek, C.A., Meyer, A.S., Silliman, K., and Kirk, L.I. 1998.
Commercial grape juice inhibits the in vitro oxidation of human l­ ow-density
lipoprotein. Journal of Agricultural and Food Chemistry 46, 834–838.
Fritsch, R.M., and Keusgen, M. 2006. Occurrence and taxonomic sig-
nificance of cysteine sulphoxides in the genus Allium L. (Alliaceae).
Phytochemistry 67, 1127–1135.

580
 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

Froehlich, O., and Schreier, P. 1986. Additional neutral volatiles from litchi
(Litchi chinensis Sonn) fruit. Flavour and Fragrance Journal 1, 149–153.
Gomez, P., Reynes, M., Dornier, M., and Hebert, J.P. 1999. La cerise des Antille:
Une exceptionnelle source de vitamine C naturelle. Fruits 54, 247–260.
Hemmerlin, A., Hoeffler, J.F., Meyer, O., Tritsch, D., Kagan, I.A.,
Grosdemange-Billiard, C., Rohmer, M., and Bach, T.J. 2003. Cross-
talk between the cytosolic mevalonate and the plastidial methyleryth-
ritol phosphate pathways in tobacco bright yellow-2 cells. Journal of
Biological Chemistry 278, 26666–26676.
Homatidou, V.H., Karvouni, S.S., Dourtoglou, V.C., and Poulos, C.N. 1992.
Determination of total volatile components of Cucumis melo L. var.
Cantaloupensis. Journal of Agricultural and Food Chemistry 40, 1385–1388.
Iranshahi, M. 2012. A review of volatile sulphur-containing compounds from
terrestrial plants: biosynthesis, distribution and analytical methods.
Journal of Essential Oil Research 24, 393–434.
Jaswir, I., Che Man, Y., Selamat, J., Ahmad, F., and Sugisawa, H. 2005. Effect of
processing conditions and storage on retention of volatile components
of durian leather. Journal of Food, Agriculture and Environment 3, 65–71.
Johnston, J.C., Welch, R.C., and Hunter, G.L.K. 1980. Volatile constituents of
litchi (Litchi chinesis Sonn). Journal of Agricultural and Food Chemistry
28, 859–861.
Jordan, M.J., Shaw, P.E., and Goodner, K.L. 2001. Volatile components in aque-
ous essence and fresh fruit of Cucumis melo cv. Athena (muskmelon)
by GC-MS and GC-O. Journal of Agricultural and Food Chemistry 49,
5929–5933.
Keay, R.W. 1989. Trees of Nigeria. In A Revised Version of Nigerian Trees,
Keay, R.W., Onoche, C.F.A., and Stanfield, D.P. (eds.). Clarendon Press,
Oxford, 476.
Klotzbach, T. 1995. World market for fresh lychees. Rap Market Information
Bulletin 6. http://www.milcom.com/rap/mps/lychee.html.
Laohakunjit, N., Kerdchoechuen, O., Matta, F.B., Silva, J.L., and Holmes, W.E.
2006. Postharvest survey of volatile compounds in five tropical fruits
using headspace solid phase microextraction. Asian Pacific Biotech
News, September 16.
Lasekan, O., and Ng, S.S. 2015. Key volatile aroma compounds of three black
velvet-tamarind (Dialium) fruit species. Food Chemistry 168, 561–565.
Lasekan, O., Khatib, A., Juhari, H., Patiram, P., and Lasekan, A. 2013.
Headspace solid-phase microextraction gas chromatography-mass
spectrometry determination of volatile compounds in different varieties
of African star apple fruit (Chrysophillum albidum). Food Chemistry 141,
2089–2097.
MacLeod, G., and Ames, J.M. 1990. Volatile components of star fruit.
Phytochemistry 29, 165–172.
MacLeod, G., and Pieris, N.M. 1981. Volatile flavour components of soursop
(Annona muricata). Journal of Agricultural and Food Chemistry 29,
488–490.

581
P OST H AR V EST RI P ENING P H Y SIOLOG Y

MacLeod, G., and Pieris, N.M. 1982. Volatile flavour components of mango-
steen (Garcinia mangostana). Phytochemistry 21, 117–119.
MacLeod, A.J., and Troconis, N.G. 1982. Volatile flavour components of sapo-
dilla fruit (Achras sapota L.). Journal of Agricultural and Food Chemistry
30, 515–517.
Maia, J.G.S., Andrade, E.H.A., and Zoghbit, M.B. 2000. Volatile constituents
of the leaves, fruits and flowers of cashew (Anacardium occidentale L).
Journal of Food Composition and Analyses 13, 227–232.
Maia, J.G.S., Andrade, E.H.A., and Zoghbit, M.B. 2004. Aroma volatiles from
two fruit varieties of jackfruit (Artocarpus heterophyllus Lam.). Food
Chemistry 85, 195–197.
Marin, R., Apel, M.A., Limberger, R.P., Raseira, M.C.B., Pereira, J.F.M.,
Zuanazzi, J.A.S., and Henriques, A.T. 2008. Volatile components and
antioxidant activity from some myrtaceous fruits cultivated in southern
Brazil. Latin American Journal of Pharmacy 27, 172–177.
Marostica, M.R., and Pastore, G.M. 2007. Tropical fruit flavour. In Flavour
and Fragrance, Berge, R.G. (ed.). Springer, Berlin, 189–201.
Mathesis, J.P., Buchanan, D.A., and Fellman, J.K. 1992. Volatile compounds
emitted by sweet cherries (Prunus avium cv. Bing) during fruit devel-
opment and ripening. Journal of Agricultural and Food Chemistry 40,
471–474.
McGorrin, R.J. 2007. Character-impact flavour compounds. In Sensory
Directed Flavour Analysis, Marsili, R. (ed.). Taylor & Francis, Boca
Raton, FL, 223–267.
Mussinan, C.J., and Keelan, M.E. 1994. Sulphur compounds. In Foods,
Keelan, M. (ed.). Vol. 565. American Chemical Society, Washington,
DC, 101–109.
Naf, R., and Velluz, A. 1996. Sulphur compounds and some uncommon esters
in durian (Durio zibethinus Murr). Flavour and Fragrance Journal 11,
295–303.
Nagegowda, D.A. 2010. Plant volatile terpenoid metabolism: Biosynthetic
genes, transcriptional regulation and subcellular compartmentation.
FEBS Letters 584, 2965–2973.
Newman, J.D., and Chappell, J. 1999. Isoprenoid biosynthesis in plants:
Carbon partitioning within the cytoplasmic pathway. Critical Reviews in
Biochemistry and Molecular Biology 34, 95–106.
Nijssen, L.M., Visscher, C.A., Maarse, H., Willemsens, L.C., and Boelens,
M.H. 1996. Volatile Compounds in Food: Qualitative and Quantitative
Data. 7th ed. TNO Nutrition and Food Research Institute, Zeist, The
Netherlands.
Oliveira, A.L., Lopes, R.B., Cabral, F.A., and Eberlin, M.N. 2006. Volatile com-
pounds from pitanga fruit (Eugenia uniflora L). Food Chemistry 99, 1–5.
Ong, P.K.C., and Acree, T.E. 1998. Gas chromatography/olfactory analy-
sis of lychee (Litchi chinensis Sonn). Journal of Agricultural and Food
Chemistry 46, 2282–2286.

582
 BIOS Y NT H ESIS O F F LA V OR AN D AROMA C OM P OUN D S

Perez, A.G., Olias, R., Lucaces, P., and Sanz, C. 2002. Biosynthesis of straw-
berry aroma compounds through amino acid metabolism. Journal of
Agricultural and Food Chemistry 50, 4037–4042.
Perez, A.G., Olias, R., Sanz, C., and Olias, J.M. 1992. Aroma components
and free amino acids in strawberry variety Chandler during ripening.
Journal of Agricultural and Food Chemistry 40, 2232–2235.
Pino, J.A., Marbot, R., Rosado, A., and Vazquez, C. 2003. Volatile constitu-
ents of fruits of Garcinia dulcis Kurz from Cuba. Flavour and Fragrance
Journal 18, 271–274.
Pino, J.A., Marbot, R., and Vazquez, C. 2002. Characterization of volatiles
in Costa Rican guava (Psidium friedrichsthalianum) fruit. Journal of
Agricultural and Food Chemistry 50, 6023–6026.
Pinto, J.A., Guedes, C.M., Moreira, R.T.A., and De Maria, C.A.B. 2006. Volatile
constituents from headspace and aqueous solution of genipap (Genipa
americana) fruit isolated by the solid-phase extraction method. Flavour
and Fragrance Journal 21, 488–491.
Rasmussen, P. 1983. Identification of volatile components of jackfruit by
gas chromatography/mass spectrometry with two different columns.
Analytical Chemistry 85, 1331–1335.
Rohmer, M., Knani, M., Simonin, P., Sutter, B., and Sahm, H. 1993. Isoprenoid
biosynthesis in bacteria: a novel pathway for the early steps leading to
isopentenyl diphosphate. Biochemistry Journal 295, 517–524.
Rowan, D.D., Allen, J.M., Fielder, S., and Hunt, M. 1999. Biosynthesis of
straight-chain ester volatiles in red delicious and granny smith apples
using deuterium-labelled precursors. Journal of Agricultural and Food
Chemistry 47, 2553–2562.
Sampaio, T.S., and Nogueira, P.C.L. 2006. Volatile components of mangaba
fruit (Hancornia speciosa Gomes) at three stages of maturity. Food
Chemistry 95, 606–610.
Schapoval, E.E.S., Silveira, S.M., Miranda, M.L., Alice, C.B., and Henriques,
A.T. 1994. Evaluation of some pharmacological activities of Eugenia uni-
flora L. Journal of Ethnopharmacy 44, 137–142.
Schippa, C., George, G., and Fellous, R. 1993. Constituents volatils de
l’acerola. Parfums, Cosmetiques, Aromes 113, 81–84.
Schwab, W., Davidovich-Rikanati, R., and Lewinsohn, E. 2008. Biosynthesis of
plant-derived flavour compounds. Plant Journal 54, 712–732.
Shalit, M., Katzir, N., and Tadmor, Y. 2001. Acetyl-CoA: alcohol acetyltrans-
ferase activity and aroma formation in ripening melon fruits. Journal of
Agricultural and Food Chemistry 49, 794–799.
Song, J., and Forney, C.F. 2008. Flavour volatile production and regulation in
fruit. Canadian Journal of Plant Science 88, 537–550.
Supriyadi, S., Suzuki, M., Yoshida, K., Muto, T., Fujita, A., and Watanabe, N.L.
2002. Changes in the volatile compounds and in the chemical and physi-
cal properties of snake fruit (Salacca eduli Reinw) cv. Pondoh during
maturation. Journal of Agricultural and Food Chemistry 50, 7627–7633.

583
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Toulemonde, B., and Beauverd, D. 1985. Headspace analysis: Trap desorp-

tion by microwave energy application to the volatile components of
some tropical fruits. Developments in Food Science 10, 533–548.
Tsuneya, T., Ishihara, M., Shiota, H., and Shiga, M. 1980. Isolation and iden-
tification of novel terpene lactones from quince fruit (Cydonia oblonga
Mill. Marmelo). Agricultural and Biological Chemistry 44, 957–958.
Umme, A., Salmah, Y., Jamilah, B., and Asbi, B.A. 1999. Microbial and enzy-
matic changes in natural soursop puree during storage. Food Chemistry
65, 315–322.
Van Straten, S., Maarse, H., de Beauveser, J.C., and Visscher, C.A. 1988.
Volatile Compounds in Foods. 5th ed. TNO Nutrition and Food Research
Institute, Zeist, The Netherlands.
Vieira-Neto, R.D. 2002. Mangaba (Hancornia speciosa Gomes). In Frutiferas
potenciais para os tabuleiros costeiros e baixadas litoraneas, Vieira, R.D.
(ed.). Embrapa Tubuleiros Costeiros, Emdagro, Aracaju, 117–140.
Wagner, K.H., and Elmadfa, I. 2003. Biological relevance of terpenoids:
Overview focusing on mono-, di- and tetraterpenes. Annals of Nutrition
and Metabolism 47, 95–106.
Wang, Y., Wyllie, S.G., and Leach, D.N. 1996. Chemical changes during the
development and ripening of the fruit Cucumis melo (cv. Makdimon).
Journal of Agricultural and Food Chemistry 44, 210–216.
Weenen, H., Koolhaas, W.E., and Apriyantoo, A. 1996. Sulphur containing
volatiles of durian fruits (Durio zibethinus Murr). Journal of Agricultural
and Food Chemistry 44, 3291–3293.
Werkhoff, P., Guntert, M., Krammer, G., Sommer, H., and Kaulen, J. 1998.
Vacuum headspace method in aroma research: flavour chemistry of yellow
passion fruits. Journal of Agricultural and Food Chemistry 46, 1076–1093.
Willson, C.W., Shaw, P.E., Knight, R.J., Nagy, S., and Kim, M. 1985. Volatile con-
stituents of carambola (Averrhoa carambola L). Journal of Agricultural
and Food Chemistry 33, 199–201.
Wu, Y., Pan, Q., Qu, W., and Duan, C. 2009. Comparison of volatile profiles
of nine litchi (Litchi chinensis Sonn.) cultivars from Southern China.
Journal of Agricultural & Food Chemistry 57(20), 9676–9681.
Wyllie, S.G., and Leach, D.N. 1992. Sulphur-containing compounds in the
aroma volatiles of melon (Cucumis melo). Journal of Agricultural and
Food Chemistry 40, 253–256.
Wyllie, S.G., Leach, D.N., Wang, Y., and Shewfelt, R.L. 1994. Sulphur volatiles
in Cucumis melo makdimon (muskmelon) aroma: Sensory evaluation
by gas chromatography-olfactrometry. In Sulfur Compounds in Foods,
Mussinan, C.J., and Keelan, M.E. (eds.). ACS Symposium Series 564.
American Chemical Society, Washington, DC, 36–48.
Zabetakis, I., and Holden, M.A. 1997. Strawberry flavour: Analysis and bio-
synthesis. Journal of the Science of Food and Agriculture 74, 421–434.

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Chapter 18

Impact of Postharvest
Technologies on the
Flavor of Fresh Fruits
and Vegetables
Charles F. Forney
Agriculture and Agri-Food Canada,
Kentville, Nova Scotia, Canada

Abstract 586
18.1 Introduction 587
18.2 Flavor of Fruits and Vegetables 588
18.2.1  Sensory Assessment 588
18.2.2  Chemical Flavor Constituents 589
18.3 Mechanism of Flavor Change 590
18.3.1  Metabolic Changes 591
18.3.2  Diffusional Changes 592
18.4 Impact of Postharvest Technologies 593
18.4.1  Ripening Manipulation 593
18.4.2  Temperature and Cold Storage 595
18.4.3  Controlled Atmosphere Storage 597
18.4.4 Packaging 599

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18.4.5  Edible Coatings 601

18.4.6  Postharvest Treatments 601
18.4.6.1 Cutting 602
18.4.6.2  Heat Treatments 603
18.4.6.3 Irradiation 604
18.4.6.4 Ozone 605
18.4.6.5  Chemical Fumigation 606
18.5 Flavor Enhancement 607
18.6 Conclusions and Future Perspectives 607
References 608

Abstract
The flavor of fruits and vegetables is complex and dynamic and is affected
by many pre- and postharvest factors. Desirable flavor is a critical factor
that can determine consumer satisfaction and thus sustainability of mar-
kets. While postharvest technologies are designed to prevent decay and
breakdown and maintain good appearance of fresh produce, their effect
on flavor is often neglected. Flavor is the human perception of a complex
combination of volatile, nonvolatile, and structural components contribut-
ing to appearance, aroma, taste, and texture. Volatile compounds contrib-
uting to flavor consist of diverse chemistries, including esters, terpenes,
alcohols, aldehydes, ketones, lactones, and sulfur compounds. Nonvolatile
compounds contributing to flavor include sugars, acids, and phenolics.
Changes in flavor can result from metabolic changes in flavor compounds,
diffusional loss of volatile flavor compounds, or gains of compounds from
the storage environment or treatments. Harvest maturity and posthar-
vest manipulation of ripening are important for optimizing flavor. The
postharvest environment, including temperature, atmosphere modifica-
tion through controlled atmosphere (CA) storage, modified atmosphere
packaging (MAP) or coatings, and the application of various postharvest
treatments, can preserve, enhance, or degrade product flavor. Reduced
temperatures can slow metabolic changes and diffusional loss of flavor
compounds, but also may inhibit normal ripening and cause flavor loss in
some chilling-sensitive commodities. Low oxygen atmospheres can inhibit
flavor synthesis or induce anaerobic atmospheres that can cause fermenta-
tion, which can result in off-flavors. Cutting fresh produce to add value and
consumer convenience can alter flavor by inducing the production of sec-
ondary flavor compounds, altering metabolism, and increasing diffusional
losses. A variety of postharvest treatments, such as heat, irradiation, ozone,
and chemical fumigation, are used to reduce decay, eliminate quarantine
pests, ensure product safety, and prolong storage life. These treatments

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may affect flavor by altering product physiology. Opportunities also exist

to enhance product flavor by use of high-flavored cultivars, optimization
of fruit ripening, design of flavor-retaining packages and coating formula-
tions, and optimization of technologies to enhance produce flavor.

18.1 Introduction
Flavor is an important characteristic determining quality and consumer
satisfaction of fresh fruits and vegetables. However, the flavor of fruits
and vegetables often does not meet the expectation of consumers. Initial
purchases are typically based on product appearance and firmness, while
repeat purchases depend on internal quality factors such as texture and
flavor (Baldwin, 2002). Loss of flavor that occurs during postharvest han-
dling, storage, and marketing of fresh fruits and vegetables often precedes
the loss of visual quality (Gorny, 2005). Therefore, ensuring good flavor
is critical to increase consumption and maintain and expand markets for
fresh produce.
A wide array of technologies have been developed to prolong the mar-
ket life of whole and fresh-cut produce. However, due to the diversity of
fruits and vegetables, postharvest technologies tend to be commodity spe-
cific in order to accommodate physiological and morphological differences.
Diversity among fresh produce is reflected in the variability of factors, such
as size, shape, firmness, cuticle thickness, transpiration rate, maturity, rip-
ening physiology, chilling sensitivity, metabolic activity, and susceptibility
to decay. These factors dictate how the product must be handled to opti-
mize market life and quality and will determine the appropriate postharvest
technology required. As a result, a wide range of strategies and technolo-
gies to prolong market life have been developed. Manipulation of product
maturity and ripeness can be achieved through harvest timing, as well as
a variety of postharvest treatments that can inhibit or stimulate ripening.
Proper precooling and postharvest temperature management slow meta-
bolic activity and product deterioration. Atmosphere modification through
controlled atmosphere (CA) technologies or modified atmosphere packag-
ing (MAP) during storage, transportation, and marketing can slow senes-
cence, inhibit decay, and reduce dehydration. Edible coatings can act as
diffusional barriers to gas exchange and have effects on produce market
life similar to those seen with CA and MAP. A variety of chemical and physi-
cal treatments have also been developed to reduce decay, inhibit microbial
growth, and slow deterioration.
Most of these postharvest technologies have been developed and opti-
mized to maintain visual quality and extend shelf life, while consideration
of organoleptic quality characteristics that are critical for consumer satis-
faction has often been neglected. Postharvest flavor change can be affected

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by many pre- and postharvest factors that are typically manipulated to

extend market life (Baldwin et al., 2007). However, the impact of many of
these factors and postharvest technologies on product flavor is not always
appreciated or understood. Therefore, in this chapter, the basis for product
flavor, the basic mechanisms responsible for postharvest flavor change in
fresh produce, and the potential impacts of various postharvest technolo-
gies on flavor are addressed.

18.2  Flavor of Fruits and Vegetables

Flavor is a human perception of a complex combination of volatile, nonvola-
tile, and structural components contributing to appearance, aroma, taste,
and texture (Drewnowski, 1997). Volatile compounds are primarily respon-
sible for the unique flavor characteristics that distinguish different fruits
and vegetables and determine their desirability to the consumer. Volatile
aroma compounds are detected by olfactory nerve endings in the olfactory
epithelium, which have a wide range of sensitivity, depending on the com-
pound (Holley, 2006). Signals from these receptors are sent to the brain
where they are processed, giving a perception of aroma and contributing to
flavor perception (Baldwin et al., 2007). Smelling volatile compounds from
produce through the nose (orthonasal) may provide a different perception
than when the aroma is perceived during chewing, where the aroma goes
to the olfactory bulb via the back of the throat (retronasal) (Voirol and
Daget, 1987). Nonvolatile compounds contribute to the taste of the prod-
uct and influence the perception of volatile compounds (Salle, 2006). The
taste of nonvolatile compounds is sensed by receptors on the tongue, which
are found in the taste buds. Compounds such as sugars, acids, salts, and
glucosides and alkaloids illicit responses from these receptors and are per-
ceived as sweet, sour, salty, and bitter, respectively. Food properties, such
as temperature, viscosity, and polarity, can affect vapor pressure and aroma
release in the mouth (Taylor and Linforth, 1994). Product textural changes
can also affect the olfactory perception of volatile compounds in a food
product (Lubbers, 2006). Texture changes such as the development of mea-
liness affects juiciness, which can affect flavor perception (Baldwin, 2002).
Texture is perceived by both the tactile and auditory senses. Perceived fla-
vor is therefore an integration of all sensory inputs, including visual appear-
ance, taste, olfactory responses, tactile responses, and auditory cues.

18.2.1  Sensory Assessment

The measurement of human perception of flavor by sensory assessment
is complex and challenging. While the analysis of chemical compounds

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responsible for flavor can generate a lot of data on sugars, acids, sugar/
acid ratios, and aroma volatiles, these data mean little if they are not related
to the human perception of flavor. Sensory tests are designed to answer a
variety of different questions, which include basic information about flavor,
differences between samples or products, magnitude of differences, and
consumer preference or “likeability.” Sensory tests can run from objective
to subjective (Baldwin et al., 2007).
Fundamental flavor information can be determined using a trained
panel to determine different flavor aspects and descriptors. Trained panels
can objectively rate the intensity of different flavor aspects and, in this way,
act like an instrument providing objective measures. Trained panels are
used to understand what comprises flavor, texture, and aroma in a quan-
titative manner (Meilgaard et al., 1999). Consumer panels, on the other
hand, are large, untrained panels that can rate the “likeability” of an attri-
bute or the “preference” of one product over another, or rank samples for a
particular attribute, such as sweetness (Baldwin et al., 2007). Flavor per-
ception is influenced by the socioeconomic, ethnic, and geographical back-
ground of the Panellists, further complicating sensory flavor e­ valuation
(Baldwin, 2002).
Sensory studies for fruits and vegetables have been used to iden-
tify optimal harvest maturity (Tandon et al., 2000), evaluate flavor qual-
ity in breeding material (Baldwin et al., 1998), determine optimal storage
(Maul et al., 2000) and handling conditions (Bett et al., 2001; Hagenmaier
and Shaw, 2002; Bai et al., 2003), assess effects of disinfestation or pre-
conditioning techniques on flavor quality (Bai et al., 2004; Plotto et al.,
2006), and measure flavor quality over the postharvest life of the product
(Baldwin, 2002).

18.2.2  Chemical Flavor Constituents

In addition to sensory analysis, chemical constituents that impact flavor
can be characterized through various analytical methods. Gas chroma-
tography is typically used to analyze volatile compounds and has identi-
fied more than 6900 different volatile compounds in food products (Misry
et al., 1997). These compounds represent a diverse range of chemistries,
including esters, terpenes, alcohols, aldehydes, ketones, lactones, and sul-
fur compounds (Song and Forney, 2008). Each commodity has a unique
volatile profile, which varies within a species, depending on maturity,
cultivar, and other factors (Baldwin et al., 2007). In addition to primary
volatile compounds that are present in fruits and vegetables, secondary
volatile compounds are formed when plant tissues are disrupted. Physical
actions such as bruising, cutting, chewing, freezing, and heating can result
in cell rupture, the mixing of enzymes and substrates, and physiological

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responses to stress that result in the production of a variety of compounds,

many of which contribute to flavor (Beaulieu and Baldwin, 2002). Onions
(Allium cepa L.), for example, produce a variety of reactive organosulfur
flavor compounds when S-alk(en)yl cysteine sulfoxides are hydrolyzed
by the enzyme alliinase upon cellular disruption (Järvenpää et al., 1998;
Randle and Lancaster, 2002). Also, physical disruption of plant tissues can
catalyze the formation of volatiles that are products of the lipoxygenase
and hydroperoxide lyase pathways (Riley and Thompson, 1998; Myung
et al., 2006; Iyer et al., 2010). This is reflected by increases in C6 aldehydes
and alcohols, including hexanal, cis-3-hexenal, trans-2-hexenal, hexanol,
­cis-3-hexenol, and trans-2-hexenol, which can impart “green” aroma notes.
The diverse chemistry and metabolic pathways involved in the synthesis
of both primary and secondary flavor compounds create an ever-changing
flavor profile, making aroma volatile chemistry complex. There are numer-
ous reviews of different aspects of metabolic processes involved in volatile
biosynthesis (Sanz et al., 1997; Baldwin et al., 2000; Fellman et al., 2000;
Beaulieu and Baldwin, 2002).
Nonvolatile compounds also contribute to the flavor of fresh produce.
The most prominent of these are sugars and acids. Sugars and sugar alco-
hols are perceived as sweet in most fruits and some vegetables. Common
sugars found include glucose, fructose, and sucrose (Beaulieu and Baldwin,
2002). Sugar alcohols, such as sorbitol and mannitol, contribute to sweetness
in some fruit and vegetables. Sorbitol can comprise up to 10% of the total sug-
ars in sweet cherries (Prunus avium L.), but varies among c­ ultivars (Usenik
et al., 2008). In celery (Apium graveolens L.) stalks, mannitol comprises about
a third of the total carbohydrate (Rupérez and Toledano, 2003). Organic
acids are perceived as sour, and their composition varies among fruits and
vegetables. Common organic acids found include citric in citrus (Citrus sp.),
blueberries (Vaccinium sp.), and tomatoes (Solanum ­lycopersicum L.); malic
in apples (Malus sylvestris (L.) var. domestica (Borkh.) Mansf.); tartaric in
grapes (Vitis vinifera L.), and quinic in cranberry (Vaccinium macrocar-
pon Ait.) (Baldwin et al., 2007; Forney et al., 2012b). Other, less abundant
compounds can also impact flavor, such as the polyphenols (+)-catechin
and (–)-epicatechin and the polymeric flavan-3-ols, which impart bitterness
(Serra Bonvehí and Ventura Coll, 1997).

18.3  Mechanism of Flavor Change

To appreciate the potential effects of postharvest technologies on the flavor
of fresh fruits and vegetables, an understanding of what determines fla-
vor and how it changes following harvest is needed. Chemical constituents
ultimately dictate human perception of flavor. Understanding how these
chemical components change in response to postharvest technologies can

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provide insights into how product flavor can be preserved and, in some
cases, enhanced during postharvest handling and marketing.
The two primary mechanisms of flavor change in fresh produce are
metabolic and diffusional (mass transfer) (Voilley and Souchon, 2006).
Both of these processes occur during development in the field as well as
postharvest. Metabolic changes involve the synthesis or catabolism of
flavor compounds or the synthesis of off-flavor compounds. Physiological
factors, such as maturity and ripeness and response to the growing and
postharvest environments, will dictate metabolic processes that affect both
volatile and nonvolatile flavor compounds. Preharvest factors affecting fla-
vor are diverse but include water availability (Baldwin et al., 2007), fertil-
ization (Randle and Lancaster, 2002), and chemical applications (Podoski
et al., 1997). The harvest maturity of fruits and vegetables has a large effect
on the postharvest flavor, with fruit harvested underripe having limited fla-
vor potential. For example, melon (Cucumis melo L.) maturity at harvest
affects the fruit flavor, with riper fruit having a sweeter aromatic taste than
underripe fruit even after postharvest ripening (Beaulieu et al., 2004).
Volatile compounds contributing to flavor are subject to diffusional changes
in which diffusion and mass transfer of volatile compounds into and out of
the commodity occur. Diffusion is determined by the volatility of the com-
pound, its concentration gradient, and diffusional barriers in the fruit or
vegetable or packaging materials. The role of each of these mechanisms
depends on the product, the environment in which it is held, and the treat-
ments to which it is subjected.

18.3.1  Metabolic Changes

The metabolic processes that affect postharvest flavor change of fruits and
vegetables vary due to the diversity of volatile and nonvolatile compounds
responsible for the commodity’s flavor and the diverse physiology of the
commodities. In a single commodity, many metabolic pathways are involved
in producing the complex mixture of compounds responsible for its flavor.
Climacteric fruit are prime examples of commodities that actively synthe-
size flavor compounds following harvest, while nonclimacteric fruits and
vegetables have more limited postharvest flavor development (Whitaker,
2008). The postharvest environment also affects metabolic processes con-
tributing to produce flavor and includes storage temperature, atmosphere
composition, and environmental stresses, such as heat, freezing, ozone,
radiation, or chemical treatments (Song et al., 1996; Forney and Jordan,
1998; Forney et al., 2000a, 2007; Song et al., 2001). Physical handling, includ-
ing cutting of fresh fruits and vegetables to add value and convenience, may
also induce metabolic changes impacting flavor (Barry-Ryan and O’Beirne,
1998; Järvenpää et al., 1998).

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18.3.2  Diffusional Changes

Volatile compounds responsible for flavor can be lost through diffusion,
which is determined by the volatility of the compound, its partition-
ing coefficient in the product matrix, diffusional barriers within and
surrounding the product, and the concentration gradient between the
product and the surrounding atmosphere (Voilley and Souchon, 2006).
The concentration gradient can be affected by packaging and storage
conditions. For fruits and vegetables that have effective intact natu-
ral barriers, the diffusional loss of flavor volatiles should be relatively
slow. However, in fresh-cut products where natural barriers have been
removed through peeling and cutting, diffusional loss may be signifi-
cant (Forney, 2008). Since fresh produce is living and responding to its
environment, permeation rates and volatile concentrations may change
during storage due to metabolic and physiological processes, making it
difficult to determine whether flavor changes are a result of diffusion or
metabolism.
The diffusional losses of flavor volatiles in fresh-cut produce have
been demonstrated. In several studies where fruit tissue was thinly sliced,
creating a very high surface-to-volume ratio, which would enhance diffu-
sional loss, loss of volatile esters after 24 h of storage at 4°C was about
80% in cantaloupe (Cucumis melo L.) (Beaulieu, 2007) and 50% in pineapple
(Ananas comosus (L.) Merr.) (Lamikanra and Richard, 2004). In another
study that considered diffusion as a mechanism of flavor loss, rapid loss
of low-molecular-weight esters (< C8) from thin-sliced cantaloupe held in
open or closed petri dishes was attributed to off-gassing due to the high
vapor pressures and low boiling points of the esters (Beaulieu, 2007). There
was no evidence for catabolic loss through esterase-driven ester loss since
there was a loss of esterase activity and no increase of products of esterase-
mediated ester metabolism.
In addition to the loss of volatile compounds through diffusion, unde-
sirable volatile compounds can diffuse into the fresh product from the stor-
age environment or packaging materials, resulting in the development of
off-odors and -flavors. Undesirable odors can originate from a variety of
sources, including chemicals, building materials, packaging, microorgan-
isms (fungi, bacteria), and other produce (Baigrie, 2003). Diffusion of these
compounds into fresh fruits or vegetables is dependent on their concentra-
tion, chemistry (i.e., the partitioning coefficient in the product matrix), and
exposure time. The accumulation of these off-odor compounds and the loss
of desirable flavor compounds during postharvest storage and handling
may contribute to the development of “stored” or “old” flavors in fresh pro-
duce (Forney et al., 2009).

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18.4  Impact of Postharvest Technologies

18.4.1  Ripening Manipulation

The flavor of fruit is dependent on its degree of maturation and ripeness;
however, associated with fruit flavor development is softening and senes-
cence, making the fruit more susceptible to physical damage, decay, and
reduced market life. Harvesting fruit underripe and delaying fruit ripen-
ing are therefore used to prolong storage and market life. The primary
methods of ripening manipulation include early harvest, rapid cooling and
temperature management, atmosphere modification, and treatments with
ethylene inhibitors.
Harvest maturity of fruit is a compromise between maximizing
the market life and optimizing the eating quality (Eccher Zerbini, 2008).
Immature and underripe fruit are firmer and more resistant to bruising
and decay than ripe fruit. However, immature and underripe fruit may
never develop optimal flavor due to a low sugar and high acid content and
a reduced ability to produce aroma volatiles. Sugar content increases as
fruit ripens on the plant, but changes after harvest are dependent on spe-
cies, whereas acid content tends to decrease during ripening both before
and after harvest in most species. Aroma volatile composition changes
both quantitatively and qualitatively during ripening, and postharvest com-
position is dependent on harvest maturity, fruit physiology, and ripening
or storage conditions. Nonclimacteric fruit have limited postharvest flavor
development. For example, strawberry (Fragaria × ananassa Duchesne)
fruit harvested half red develop a full red color postharvest, but remain
firm and tart and lack the full-ripe strawberry aroma (Forney et al., 2000b).
Other non- or weakly climacteric fruit, such as oranges (Citrus sinensis
L.) or highbush blueberries (Vaccinium corymbosum L.), also demonstrate
minimal postharvest flavor development (Baldwin et al., 1995; Forney et al.,
2001). On the other hand, climacteric fruit harvested physiologically mature
can develop acceptable flavor through postharvest ripening; however, full
flavor may never be achieved. Apples harvested immature had lower volatile
production than later harvested fruit (Vanoli et al., 1995), which resulted in
a lack of flavor development (Mattheis, 1991). When tomatoes were har-
vested mature green and then ripened at 20°C, they had less fruity aroma
and sweetness and more sourness and off-flavor than fruit harvested ripe
(Kader et al., 1978). However, fruit harvested light pink and then ripened
at 20°C had less fruity aroma, but similar sweetness, sourness, and off-fla-
vor as ripe harvested fruit. Mango (Mangifera indica L.) fruit harvested at
the postclimacteric green maturity stage and ripened postharvest at 21°C
developed similar firmness, sugar content, and color as fruit harvested

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ripe, but had a higher acid content. In addition, the postharvest ripened
fruit had higher total volatiles than fruit harvested ripe, with the highest
levels of monoterpenes, sesquiterpenes, and aromatic compounds, whereas
the ripe harvested fruit had the highest concentration of esters (Lalel et al.,
2003). These compositional differences suggest a difference in fruit flavor.
Climacteric fruits are responsive to the ripening hormone ethylene,
which stimulates ripening and flavor development when endogenously
produced or exogenously applied. Therefore, a common strategy is to har-
vest climacteric fruit, such as banana (Musa × paradisiaca L.) and tomato,
unripe and firm. Handling and shipping firm fruit reduces damage that can
occur when shipping fruit long distances. When fruit reaches its market,
ripening can then be stimulated with ethylene to achieve an acceptable fla-
vor, texture, and eating quality (Lèlievre et al., 1997). Commercial ripening
rooms that can control ethylene concentration, temperature, and humidity
are typically used for this purpose. The flavor quality reached under these
conditions is dependent on the harvest maturity and fruit species. Many
underripe fruits may not be able to obtain full flavor through postharvest
ripening. In tomato, fruit harvested immature green produced ripe fruit
with lower aroma volatile levels than mature-green harvested fruit, while
fruit harvested at the table-ripe stage had the highest intensity of flavor
components (Watada and Aulenbach, 1979). Ethylene treatment of toma-
toes was reported to speed ripening but did not affect flavor when compared
to fruit ripened without exogenous ethylene (Kader et al., 1978). The matu-
rity stage at which apple fruit are harvested also affects ester formation
and acid content during postharvest ripening (Fellman et al., 2000), thus
affecting fruit flavor. Flavor change during postharvest ripening of apples
during cold storage varies substantially among cultivars; therefore, the rate
and nature of flavor change can be cultivar specific (Corollaro et al., 2013).
Inhibition of ethylene action has been effective in prolonging the stor-
age life of apples and other fruits and vegetables. This has been achieved
through the application of 1-methylcyclopropene (1-MCP), which blocks
ethylene-induced ripening and other physiological processes (Blankenship
and Dole, 2003). Application of 1-MCP can be achieved through posthar-
vest fumigation (SmartFresh™ ) or, more recently, as a preharvest spray
(Harvista™ ). These treatments are effective in maintaining fruit firmness
and inhibiting acid loss, color change, and volatile synthesis (Blankenship
and Dole, 2003). In addition, they are effective in delaying senescence and
extending the storage life of apples and many other fruits and vegetables
(Watkins, 2006). The 1-MCP-induced reduction in volatile production
has been reported in many climacteric fruits, including banana (Golding
et  al., 1998), plums (Prunus domestica) (Abdi et al., 1998), apples (Fan
and Mattheis, 1999; Mattheis et al., 2005), and pears (Pyrus communis L.)
(Argenta et al., 2003). However, in apples, this reduction in volatile produc-
tion by 1-MCP may not be perceived as negatively impacting consumer

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preference. ‘Anna’ and ‘Gala’ apple fruit treated with 1-MCP had reduced
ester production and perceived fruity aroma, but were preferred over con-
trols, possibly due to textural differences (Lurie et al., 2002; Maria et al.,
2007). Treatments with 1-MCP were also shown to inhibit ethylene-induced
bitterness in fresh carrots, which is caused by accumulation of isocouma-
rin-6-methyoxymellein (Fan and Mattheis, 2000; Song et al., 2003).

18.4.2  Temperature and Cold Storage

The management of produce temperature after harvest is critical for man-
aging product flavor and overall quality. Temperature has a profound effect
on the rates of metabolic processes, and as temperature increases between
0°C and 30°C, the rates of respiration and metabolism increase exponen-
tially (Saltveit, 2004). Therefore, cooling fruit reduces metabolic changes
in fresh produce, including synthesis and catabolism of flavor compounds,
as well as physiological processes, such as fruit ripening. Manipulation of
temperature can be used to delay or stimulate ripening and other metabolic
activities in order to optimize product flavor. Most stone fruit (peaches, nec-
tarines, plums) are held at 2.2°C to slow ripening and flavor development,
but then are held between 10°C and 25°C to ripen and develop optimum
flavor (Crisosto, 1999). The rate of ripening depends on the temperature
and cultivar. Ripening at temperatures of >25°C may induce off-flavors.
However, storage of some cultivars at 2.2°C may result in fruit with poor
texture and flavor, which can be prevented if these fruit are preconditioned
(held at 20°C for 48 h) prior to cold storage (Lurie and Crisosto, 2005).
Pears actually require a period of cold storage before they can properly
ripen and develop desired flavor (Sugar and Basile, 2009). Cold storage
time is cultivar dependent, with ‘Bartlet’, ‘Comice’, and ‘Anjou’ requiring
15, 30, or >60 days, respectively, at –1°C to properly ripen and develop
optimum flavor at 20°C. This cold storage time can be reduced if fruit are
held at 5°C or 10°C and treated with ethylene following harvest. Mandarin
(Citrus reticulata Blanco) fruit were reported to maintain good flavor when
stored at temperatures between 5°C and 10°C, but rapidly developed off-
flavors when warmed to 20°C (Obenland et al., 2013). These off-flavors were
associated with a rise in alcohol and esters in the fruit held at 20°C.
While the quality of many fruits and vegetables is preserved at tem-
peratures near freezing (–1°C to 0°C), some are sensitive to low tempera-
tures and may develop chilling injury (Paull, 1990). Crops of tropical or
subtropical origin may be injured after a period of exposure to chilling tem-
peratures below 10°C–15°C but above their freezing points (Wang, 2004).
Some crops of temperate origin are also susceptible to chilling injury but
have lower threshold temperatures of <5°C (Bramlage and Meir, 1990).
Chilling injury can be expressed as the physiological breakdown of tissues,

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failure to ripen normally, and loss of flavor (Saltveit and Morris, 1990).
When ripe tomato fruit were held for 2 days at the chilling temperatures
of 5°C, 10°C, and 12.5°C, their ripe aroma, sweetness, and tomato flavor
were rated by a sensory panel as significantly lower than those of tomatoes
held at 20°C (Maul et al., 2000). Similarly, when mature-green or light pink
tomatoes were held at 5°C for 7 days prior to ripening at 20°C, they were
found to be more acidic than similarly ripened fruit that were not held at 5°C
(Kader et al., 1978). These reductions in sensory ratings were accompanied
by reductions in volatile and nonvolatile flavor components. Some cultivars
of plums are chilling sensitive and will develop internal browning and poor
flavor if held at 1°C–5°C (de Kock and Taylor, 2014). However, to develop
optimum flavor, they must be cooled to –0.5°C for 3–10 days prior to ripen-
ing at 7.5°C. Similarly, fruit of chilling-sensitive peach cultivars that were
held at 5°C followed by 3 days at 20°C had a reduction in the lactones that
were shown to be aroma impact compounds of peach (Xi et al., 2012). This
chilling-reduced production of lactones was partially ameliorated by hold-
ing fruit at 0°C rather than 5°C, or by subjecting them to a low-temperature
conditioning treatment (8°C for 3 days followed by storage at 5°C).
In addition to the effects on metabolic activity, the volatility of volatile
flavor compounds increases as temperature increases. Therefore, elevated
temperatures can speed up both quantitative and qualitative changes in
flavor compounds, thus affecting flavor. To preserve flavor, it is thought
that the lowest acceptable storage temperature is generally best, espe-
cially when fruit are harvested at optimal eating quality. However, with the
dynamic and complex nature of flavor components, changes in temperature
can affect rates of both synthesis and loss of flavor compounds, and these
effects can be compound specific. In stored strawberry fruit, a change in
ester composition was found to be temperature dependent. When straw-
berry fruit were stored at 1°C, concentrations of the esters ethyl butanoate
and ethyl hexanoate increased, while those of methyl butanoate and methyl
hexanoate increased during storage at 15°C (Forney et al., 2000b). The tem-
perature-dependent effects on volatile composition could affect fruit fl ­ avor
characteristics in strawberry as well as other fruits and deserve further
assessment.
Extended cold storage can also result in the development of off-­
flavors, which are often described as old or stored. This may be the result
of the loss of natural flavor compounds or the migration of off-odor com-
pounds into the product from the storage environment. It is recognized
that flavor can be lost during the storage of fresh fruits and vegetables,
and that this may occur prior to a decline in product appearance (Baldwin
et al., 2007). The appearance of fresh strawberries was maintained for
7–9 days when stored at 5°C, but acceptable flavor was lost several days
sooner, depending on cultivar (Pelayo et al., 2003). Similar results were
reported for tomato (Maul et al., 2000) and mandarins (Tietel et al., 2012).

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Depending on the chemistry of the volatile compounds found in the stor-

age ­environment, many can migrate into fresh fruits or vegetables. In
situations with mixed storage of incompatible commodities, apples and
pears can develop an earthy off-flavor when stored with potatoes (Solanum
tuberosum  L.) (Hardenburg et  al., 1986). Off-flavors in citrus can also
develop when stored with strong scented vegetables such as onions. A
musty off-flavor that results from chloroanisole migration has been found
in a variety of foods, including raisins and potatoes (Tindale et al., 1989;
Daniels-Lake et al., 2007). This compound can be produced by microbial
methylation of chlorophenols, which are commonly found in treated lum-
ber, recycled paper products, and ­adhesives. The contribution of taints to
the loss of desirable flavor during storage has received little attention, but
could be an important factor affecting product flavor life. Efforts to prevent
the migration of off-flavor taints into fresh produce could be beneficial to
improve the flavor quality of stored produce.

18.4.3  Controlled Atmosphere Storage

Storage atmospheres with reduced concentrations of O2 or elevated levels
of CO2 can decrease rates of senescence and ripening and slow quality loss
of fresh fruits and vegetables, and thus prolong the period of consumer
acceptance (Parsons et al., 1970; Bangerth, 1973). Storage in CA reduces
chlorophyll loss, softening, and production of volatile compounds that con-
tribute to flavor. A reduction in the production of volatile compounds that
contribute to aroma and flavor has been reported in many fruit, includ-
ing apple, black currant (Ribes nigrum L.), kiwifruit (Actinidia deliciosa
[A. Chev.] C.F. Liang et A.R. Ferguson var. deliciosa), pear, and strawberry
(Forney et al., 2009). Reduced volatile production of fruit in CA storage is
enhanced with increased storage duration and decreased O2 concentration
(Streif and Bangerth, 1988; Yahia et al., 1990; Mattheis et al., 1995; López
et al., 2001; Forney et al., 2009). Volatile production can be partially to
completely restored if fruit are held in air following CA storage (Streif and
Bangerth, 1988; Yahia et al., 1990), but the amount of recovery decreases
with increased storage duration and decreased O2 concentration during
storage (Streif and Bangerth, 1988; Yahia et al., 1990).
While low O2 concentration reduces volatile production, it has vari-
able effects on fruit flavor and aroma. Sensory analysis of ‘Gala’ apples
stored 24 weeks in 1.2–5 kPa O2 resulted in fruit with greater overall accept-
ability that were more crisp, firm, juicy, and sour and had fewer off-flavors
than fruit stored in air (Cliff et al., 1998). However, in another study, ‘Gala’
apples stored 20 weeks in CA were judged to have lower fruity and floral
aroma, higher vegetative and citrus characteristics, and greater sourness
and astringency than air-stored fruit (Plotto et al., 1999). Lower amounts

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of esters were produced by ‘Fuji’ apples stored in 1 or 2 kPa O2 than by fruit

stored in air for about 30 weeks, but sensory flavor scores were similar 1
day after fruit were removed from CA (Echeverría et al., 2004). Apple fruit
flavor and acceptability are determined by a number of quality attributes,
which vary among cultivars and are influenced by the growing environ-
ment, harvest maturity, and postharvest handling, impacting consumer
preference (Daillant-Spinnler et al., 1996).
Elevated concentrations of CO2 have been shown to reduce the devel-
opment of decay in fruits. Concentrations of 10 kPa CO2 or more are par-
ticularly effective in controlling gray mold caused by Botrytis cinerea in
berry crops. Treatments with 40 kPa CO2 for 24–48 h followed by storage
in atmospheres of 12 kPa O2 and 12 kPa CO2 preserved quality of ‘Flame
Seedless’ and ‘Crimson Seedless’ grapes held at 1°C for up to 7 weeks while
having no adverse effects on flavor (Teles et al., 2014). However, off-flavors
developed in early-season ‘Thompson Seedless’ (Crisosto et al., 2002a) and
‘Redglobe’ (Crisosto et al., 2002b) grapes held in 15 kPa CO2 atmospheres.
Elevated CO2 concentrations of 10–20 kPa were also reported to prevent
chilling-induced flavor loss in peaches (Lurie and Crisosto, 2005).
Injurious levels of O2 and CO2 can induce the formation of off-flavor
in fresh fruits and vegetables, resulting in a loss of quality (Fidler and
North, 1971). In addition to gas concentration, the induction of injury is
dependent on exposure time, storage temperature, and product physiol-
ogy. Typical responses to injurious O2 conditions include the induction
of anaerobic metabolism and the accumulation of acetaldehyde and etha-
nol (Thomas and Fidler, 1933). The accumulation of these compounds
enhances the synthesis of ethyl esters (Ke et al., 1991, 1994; Larsen and
Watkins, 1995), while reducing the production of other volatile esters
(Mattheis, 1991). However, a simple relationship between ethanol, ethyl
acetate, and acetaldehyde and off-flavor has not been demonstrated due
to the complex relationships among volatile and nonvolatile components
and product flavor (Ke et al., 1991; Larsen and Watkins, 1995). Therefore,
altered volatile profiles induced by atmosphere modifications can have
little or no impact on flavor (Ke et al., 1991) or can generate objection-
able off-flavors, as seen in fruits such as apple (Fidler and North, 1971),
mango (Bender et al., 2000), strawberry (Ke et al., 1991), and papaya
(Yahia et al., 1992) or vegetables such as broccoli (Forney et al., 1991) and
other Brassica vegetables (Forney and Jordan, 1995). In contrast, short
exposures to nitrogen or low oxygen atmospheres that induce anaero-
biosis have been reported to enhance the flavor of orange (Shaw et al.,
1992), apple (Dixon and Hewett, 2000), and feijoa (Pesis et al., 1991). In
addition, concentrations of >80 kPa CO2 can decrease the astringency
of ­p ersimmons (Toye et al., 1987; Besada et al., 2013), thus improving
their flavor.

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18.4.4 Packaging
The use of MAP has increased in recent years with the increase in mar-
keting of fresh-cut fruits and vegetables (Forney and Yaganza, 2011).
Proper package design can maintain beneficial atmospheres and prolong
the market life of these highly perishable products by minimizing dehy-
dration, reducing the browning of cut surfaces, reducing decay and micro-
bial growth, and preventing product contamination (Forney et al., 2007).
Packaging can also affect product flavor through the effects of atmosphere
composition and volatile diffusion (Forney, 2008).
Atmosphere composition can affect fresh-cut produce flavor in a
manner similar to that seen in CA storage of whole fruits and vegetables.
However, due to their high perishability and the resulting short market life,
fresh-cut produce may tolerate more extreme atmospheres. Sliced ‘Gala’
apples packaged in microperforated bags that maintained a 16 kPa O2/6
kPa CO2 atmosphere had better flavor than slices packaged in a solid mul-
tilayered polyolefin film bag that maintained a 2–3 kPa O2/6 kPa CO2 atmo-
sphere (Cliff et al., 2010). The apple slices in the microperforated bags had
higher fruity aroma and flavor and perceived sweetness, which were asso-
ciated with higher volatile concentrations of estragole and straight-chain
esters. These flavor differences could be a result of the differences in O2
concentration in the package or interactions of the fruit volatiles with the
polymers in the packaging.
Since atmosphere composition is not actively controlled in MAP, pack-
ages run a greater risk of developing anaerobic atmospheres and objection-
able off-odors if exposed to temperature abuse (Forney and Yaganza, 2011).
Warming of packages increases product respiration rate, which can deplete
package O2 and create anaerobic conditions. The resulting of off-odor
development is dependent on product physiology. While broccoli produced
substantial levels of the malodorant methanethiol when held under anaero-
bic atmospheres, other fresh-cut Brassica vegetables varied substantially
in their production of methanethiol and other malodorants (Forney and
Jordan, 1999). Fresh-cut cauliflower (Botrytis group) florets, Chinese cab-
bage (Pekinensis group), and kohlrabi (Gongylodes group) tubers, while
producing ethanol, only produced trace amounts of methanethiol and other
malodorants. Therefore, these vegetables pose less of a risk of developing
objectionable odors if exposed to abusive temperatures and their result-
ing anaerobic atmospheres during marketing. Fresh-cut iceberg lettuce is
typically packaged in low O2 atmospheres to prevent cut-edge browning.
However, these atmospheres often induce fermentation, resulting in the
production of ethanol (Cameron et al., 1995; Smyth et al., 1998) and other
volatiles, including hexanal, cis-3-hexen-1-ol, β-elemene, ethyl acetate, and
dimethyl sulfide (Tudela et al., 2013). The resulting off-odor and -flavor,

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which are typically masked by salad dressing when consumed, have been
accepted by consumers. However, other technologies to prevent cut-­lettuce
browning are needed to avoid this undesirable off-flavor and provide con-
sumers with a more desirable fresh flavor. Anaerobic off-odors associ-
ated with increased levels of ethanol, acetaldehyde, and ethyl esters are
typically found in fruit held in injurious atmospheres, as seen in packaged
pomegranate arils (Caleb et al., 2013).
To mitigate anaerobic effects, active MAP approaches have been tried
where initial atmospheres of super atmospheric O2 were established. When
‘Camarosa’ strawberry fruit was held at 2°C in MAP with an initial atmo-
sphere of 80 kPa O2, acceptable flavor was not maintained during 12 days of
storage (Allende et al., 2007). During this time, package O2 concentration
was maintained above 65 kPa, but CO2 concentrations increased to about
20 kPa, which appears to have induced anaerobic metabolism and develop-
ment of associated off-flavors.
Volatile flavor compounds in fresh produce and other food products
can interact with polymer packaging materials in several ways (Forney and
Yaganza, 2011). If the polymer acts as a barrier to the loss of flavor volatiles
from the package headspace, the resulting reduced concentration gradient
between the product and the surrounding atmosphere reduces diffusional
loss from the product. Most packaging materials provide a good barrier to
diffusional loss assuming the package is well sealed and leak-free. Fruity
esters in fresh-cut cantaloupe sealed in polyethylene terephthalate (PET)
bowls increased as much as 40% after 7 days of storage at 4°C (Beaulieu,
2006), whereas a rapid loss of these esters was observed under conditions
conducive to diffusional loss (Beaulieu, 2007).
Polymer packaging may also have a direct interaction with volatile
compounds in the headspace, resulting in sorption or permeation, which
removes the volatiles from the package atmosphere and ultimately the
packaged product (Brody, 2002; Forney and Yaganza, 2011). The widely
used packaging polymers polypropylene and polyethylene have a high
affinity for volatile compounds due to their nonpolar properties, whereas
polyesters, including the bio-based polymer polylactic acid (PLA), are
more polar and have a weaker affinity for flavor volatiles (Sajilata et al.,
2007). When diced onions were sealed in PLA containers, the onions
retained a fresh-cut onion aroma for 2 weeks, while those sealed in poly-
ethylene bags lost this fresh aroma within the first 2 days (Forney et al.,
2012a). This loss of aroma was associated with a rapid loss of sulfur-
containing aroma volatiles from the onions and the package headspace.
Fresh blueberry fruit stored in sealed PLA containers were found to have
better flavor than fruit held in vented PET containers (Almenar et  al.,
2010). However, no significant effects on aroma volatiles were found
between the two package types; rather, greater water loss in the vented
PET packaged fruit appeared to be responsible for the poorer flavor.

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Understanding the properties of packaging materials in relation to flavor

volatiles could help us identify additional methods for preserving product
flavor and aroma.

18.4.5  Edible Coatings

Edible coatings can provide additional options for preserving quality and
reducing flavor loss in fresh produce. Edible coatings can delay ripening,
slow senescence, inhibit discoloration, reduce dehydration, and extend mar-
ket life (Forney et al., 2009). Formulations of edible coatings are numerous
and may contain waxes, shellac, resins, cellulose, methyl celluloses, chito-
san, maltodextrin, starch, proteins, and oil emulsions (Baldwin, 1994). The
permeation properties of the coating create a modified atmosphere in
the product. Similar to MAP, flavor can be affected by metabolic effects of
the atmosphere modification or by the barrier properties of the coating to
diffusional loss of flavor volatiles. Low solid shellac, cellulose, and carnauba
wax coatings delayed ripening and extended the shelf life of mangoes with-
out altering flavor from that of uncoated fruit, even though the shellac and
cellulose coatings caused elevated levels of ethanol (Hoa et al., 2002). In
a different study, cellulose-coated mangoes exhibited increased concen-
trations of flavor volatiles compared to uncoated controls (Baldwin et al.,
1999). Mango pieces coated with carboxy methyl cellulose or maltodextin
also had better volatile retention than uncoated fruit, but coatings of chito-
san, starch, whey protein, or soybean oil emulsion resulted in poor flavor
(Plotto et al., 2004). In citrus, wax coatings that enhance gloss, as well as
less permeable coatings with high amounts of shellac or wood rosin, have
been found to restrict gas exchange, resulting in anaerobic internal atmo-
spheres of the fruit, accumulation of ethanol, altered volatile profiles, and
the development of off-flavors (Baldwin et al., 1995). In attempts to replace
synthetic waxes with natural biodegradable edible coatings, carboxymethyl
cellulose and chitosan bilayer coatings were found to enhance citrus gloss,
but they slightly impaired flavor quality in mandarins, but not in oranges
and grapefruit (Citrus × paradise Macfad) (Arnon et al., 2014). Recently,
Aloe vera gel-based coatings were reported to prolong the shelf life of sweet
cherries, while having no adverse effects on fruit flavor (Martínez-Romero
et al., 2006).

18.4.6  Postharvest Treatments

In addition to the previously discussed technologies, many fresh fruits
and vegetables are subjected to a variety of chemical and physical treat-
ments to add convenience, delay ripening, control decay, or eliminate pests

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(quarantine requirements) in order to prolong the product life or expand

market access. Many of these treatments can impact the flavor of the
treated product. Fresh produce treatments include antimicrobial dips or
sprays, heat treatments, irradiation, and fumigation with antimicrobial or
insecticidal chemicals, to name a few. The effects of several of these treat-
ments on produce quality and flavor are discussed here.

18.4.6.1 Cutting
The fresh-cut processing of whole fresh produce can cause both metabolic
and diffusional flavor changes. Secondary aroma volatiles are formed in a
number of fruits and vegetables as a result of cellular disruption. For exam-
ple, as described previously, the cutting of onions produces thiopropanal
S-oxide, prop(en)yl thiol, aldehydes, and a variety of sulfides as a result
of the reaction of S-alk(en)yl cysteine sulfoxides with alliinase that occurs
due to cellular rupture during cutting (Järvenpää et al., 1998). Fresh onion
aroma is maximized after about 30 min, but continues to change due to
further chemical reactions of the volatiles in the headspace. New aroma
volatiles are also formed as a result of lipid oxidation that can occur as a
result of cutting. When tomatoes are cut, an increase of many compounds,
including cis-3-hexenal, hexanal, and 1-penten-3-one, occurs that impacts
the product’s flavor (Baldwin et al., 2000).
In addition to the postharvest formation of flavor compounds follow-
ing cutting, flavor loss has been associated with fresh-cut processing. In
carrots, the loss of aroma was greater when cut with a dull blade than with
a sharp blade (Barry-Ryan and O’Beirne, 1998). Carrots cut with the dull
blade also developed off-odors more rapidly, which may have been a result
of injurious levels of CO2 that developed more rapidly in packages of dull-
blade cut carrots. Cantaloupe cut with a dull cork bore also developed more
off-odors and had a slightly higher level of ethanol than fruit cut with a
sharp cork bore, although both had a similar loss of aroma (Portela and
Cantwell, 2001). Lamikanra and Richard (2002) suggested that the rapid
loss of esters in cut melon tissue is a result of “stress induced enzymatic
hydrolysis of esters.” However, activity of esterase, an enzyme that breaks
down esters, decreased 24 h after cutting (Lamikanra and Watson, 2003),
and there was no relationship observed between ester loss and synthesis of
fatty acids, aldehydes, or alcohols, which are esterase products (Lamikanra
and Richard, 2002). Subjective aroma ratings of honeydew and cantaloupe
pieces were found to decline after 6 days when the cut fruit was held at 5°C
in permeable cheesecloth-covered jars (Portela and Cantwell, 1998, 2001).
Sensory tests also determined that fresh-cut orange slices stored at 4°C in
a closed plastic box that maintained ambient atmospheres lost flavor during
5 days of storage, which was associated with a loss of flavor components

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in the fruit (Rocha et al., 1995). While flavor loss is commonly associated
with cutting, definitive studies that determine the underlying mechanisms
responsible are limited.

18.4.6.2  Heat Treatments

Heat treatments in the form of hot water, hot air, or vapor heat have been
shown to have several useful applications that include the disinfestation
of insect pests, the reduction of postharvest disorders and decay, and the
extension of storage life in a variety of fruits and vegetables (Lurie, 1998).
Heat treatments alter physiological and biochemical characteristics that
can result in the slowing of ripening of climacteric fruits and vegetables,
the sweetening of commodities by either increasing sugars or decreasing
acidity, and the prevention of storage disorders, such as superficial scald
on apples and chilling injury on subtropical fruits and vegetables (Lurie,
1998). However, there is often a fine line between treatment temperature
and duration of exposure that produces desirable results versus induces
damage to the product. In addition, factors other than treatment regimes,
such as fruit maturity and cultivar, may affect fruit response to treatments
(Fan et al., 2005b, 2011).
The effects of heat treatments on fruit flavor are varied. In addition
to their effect on sweetness, acidity, and firmness, treatments have been
shown to alter the flavor volatile composition of fruit. High-temperature
forced-air treatment caused flavor loss in navel and Valencia oranges follow-
ing 4 weeks of storage (Obenland et al., 1999). This loss of flavor was associ-
ated with a reduction in concentrations of the aromatic volatiles α-pinene,
β-myrcene, and limonene. ‘Golden Delicious’ apples held 4 days at 38°C
had a substantial reduction of total volatile esters 1 day after treatment, pro-
ducing only 6% of that produced by non-heat-treated controls (Fallik et al.,
1997). However, after 6 weeks of storage at 1°C, volatile production of the
heat-treated fruit recovered and produced levels of esters similar to those
of non-heat-treated fruit. Similar results were observed with ‘Gala’ apples,
where esters and total volatiles were reduced after 2 months of storage but
returned to control levels after 4 or 6 months (Saftner et al., 2002). Sensory
analysis of this fruit showed no difference in flavor between heat-treated
and air-stored fruit after 4 and 6 months of storage. However, heat-treated
fruit that were CA stored had half the concentration of total volatiles, but
were judged to have better fruity flavor. This may have been a result of
higher acidity in the CA-stored fruit than in the heat-treated and air-stored
fruit. The effect of varying degrees of heat stress on ester production is
dependent on fruit cultivar and maturity. Heat treatments of 4 and 8 h in
46°C air increased the ester content of immature ‘Jonagold’ apples 2.5- and
1.8-fold 24 h after treatment, but a 12 h treatment reduced ester content

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50% (Forney et al., 2011). In contrast, the ester content of ‘Cortland’ apples
subjected to the same treatments was reduced 30%–65%. The effects of heat
treatments on more mature fruit and following longer storage times tended
to decrease fruit ester content compared to untreated control fruit. Fruit
with the greatest loss of esters also developed skin and flesh browning. In
tomato fruit, a 5 min 52°C hot water dip of mature-green fruit resulted in
enhanced sensory profiles of both ‘TastiLee’ and ‘Florida-47’ fruit follow-
ing ripening at 20°C when compared to fruit dipped in 25°C water (Loayza
et al., 2012). Heat-treated fruit were considered soft with a complex sensory
profile, whereas fruit from the control were firm, sour, and bland. A 2 min
dip of sour cherries in 40°C water preserved color and texture during 17
days of storage at 4°C, but was detrimental to acceptable flavor (Ravanfar
et al., 2012). On the other hand, grapes that were subjected to a 5 min 55°C
water dip had no detrimental effect on flavor while slowing decay and prod-
uct deterioration (Sabir and Sabir, 2013).
Although many reports focus on the positive response of horticul-
tural commodities to heat treatment, high temperatures with long dura-
tions may cause tissue damage and increase decay development (Klein
and Lurie, 1992; Song et al., 2001; Fan et al., 2005a). Heat stress induces
fermentation, which may be transient in mild treatments or sustained in
more severe injurious treatments. This is associated with the production
of ethanol, acetaldehyde, and ethyl esters such as ethyl acetate. Exposure
of oranges for more than 200 min to 48.5°C forced air resulted in ethanol
synthesis that was associated with perceptible off-flavors (Obenland et al.,
1999). Similarly, treatments of mangoes at 46°C and 48°C forced air for
3–5 h resulted in acetaldehyde and ethanol accumulation (Mitcham and
McDonald, 1993). Hot water dips of 1–3 min at 52°C delayed yellowing
of broccoli held at 20°C (Forney, 1995). However, the 3 min dip caused a
“green floral” off-odor, which was associated with the production of ethanol,
cis-3-hexenol, and dimethyl trisulfide (Forney and Jordan, 1998).

18.4.6.3 Irradiation
Irradiation with ultraviolet (UV) and ionizing radiation has been used
to eliminate quarantine pests, reduce microbial load, and in some cases,
induce decay resistance. Exposure of thin slices of pineapple to UV light
caused a twofold increase of the sesquiterpenes copaene and ocimene,
which have antimicrobial activity (Lamikanra and Richard, 2004) and
woody and floral aroma notes. UV treatments also increased the produc-
tion of the terpenoids ß-ionone, ß-ionone epoxide, and dihydroactinidiolide
in fresh-cut cantaloupe, and it was suggested that these terpenoids could
be detrimental to fresh melon flavor (Beaulieu, 2007). Lamikanra et al.
(2005) reported fresh-cut cantaloupe treated with UV light (1180 mW/cm 2

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for 4 min) smelled less rancid and more fruity after 6 days of storage than
control fruit. The reduction in rancid smell was associated with a reduc-
tion in lipase activity in the treated fruit. Ionizing radiation treatments
have not been shown to have significant effects on fresh-cut product fla-
vor. Pluots (Prunus domestica × Prunus armeniaca) treated with up to 1.4
kGy of gamma irradiation were reported to have a quality similar to that
of untreated controls (Duvenhage et al., 2012). In strawberry fruit, treat-
ments of 2.0 and 2.5 kGy reduced fruit decay while maintaining acceptable
flavor (Silva et al., 2009). Romaine lettuce sealed in laminate PE bags and
irradiated with 0.15 or 0.35 kGy of gamma radiation developed off-odors
after 11 days of storage at 4°C, but the irradiation treatment had no effect
on off-odor induction (Pariasca et al., 2001).

18.4.6.4 Ozone
Ozone (O3) is a highly reactive form of oxygen with strong antimicrobial
properties. Antimicrobial treatments have been developed using ozone in
air or dissolved in water to reduce the decay of fresh produce and sanitize
water and equipment used in postharvest handling. Ozone and its reactive
products may induce many physiological changes in plant tissues, and under
high doses, cellular damage may occur, causing altered membrane perme-
ability, premature senescence, loss of pigments, and surface discoloration.
The use of ozone for postharvest sanitation and decay control of fresh fruits
and vegetables during handling and storage has been investigated for com-
mercial application on a wide variety of commodities (Forney, 2003). Ozone
has been shown to have some effect on the decay of carrots. Exposure of
carrots to 1 μl/L gaseous ozone at 10°C for 2–4 days induced resistance to
Botrytis cinerea (gray mold) but not to Sclerotinia sclerotiorum (white mold)
(Song et al., 2003; Forney et al., 2007). These treatments altered both volatile
and nonvolatile flavor compounds in the carrots. Concentrations of ethanol
and hexanal were 43 and 11 times greater, respectively, than controls imme-
diately after the 4-day exposure to 1 μl/L ozone. Terpenes, which contribute
to carrot flavor, also increased following treatment. However, these increases
in volatiles were transient, and concentration returned to control levels after
8 weeks of storage. The 4-day 1 μl/L ozone treatment also increased glucose
and fructose concentrations, reduced sucrose concentrations, and induced
the production of the phytoalexin isocoumarin-6-methoxymellien to concen-
trations that could cause bitterness in the carrots. Treatment of the ­carrots
with 1-MCP prior to ozone treatment prevented changes in sugars and
induction of isocoumarin-6-methoxymellien and Botrytis resistance, indicat-
ing that these effects were mediated by ozone-induced ethylene.
Ozone treatments can also affect the flavor of fruits. Ozone treatments
of 2.5 μl/L for 4 days followed by storage at 26°C for 12 days reduced the

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decay of papaya fruit while increasing the sensory ratings of sweetness and
texture above those of untreated controls (Ali et al., 2014). In tomato fruit
subject to 7 days of 0.15 mol/mol ozone enrichment, fruit were perceptibly
sweeter, retained their firmness, and were preferred over fruit subject to
traditional storage and transit practices (Tzortzakis et al., 2007). In straw-
berries, ozone treatments of 0.35 μl/L for 3 days at 2°C decreased the pro-
duction of esters by 35% compared to controls (Pérez et al., 1999). Ethanol
concentrations also decreased in strawberries following 2 and 4 additional
days in air at 20°C, and there was no effect on ethyl acetate or acetalde-
hyde concentrations, suggesting that ozone prevented the development of
off-flavors. Sucrose, glucose, and fructose were reduced by about 20% in
ozone-treated fruit, but after an additional 2 or 4 days in air at 20°C, these
differences were less apparent and the ozone-treated fruit actually had
higher concentrations of sucrose (Pérez et al., 1999). In cranberries, ozone
induced a faint but pleasant floral aroma (Norton et al., 1968). Treatments of
1.0 μl/L ozone for 24 h had no effect on the soluble solid content of peaches
(Ridley and Sims, 1967). Schomer and McColloch (1948) found that long-
term exposure of apples to 3.25 μl/L ozone resulted in a loss of flavor.

18.4.6.5  Chemical Fumigation

In attempts to reduce the postharvest decay of fruits and vegetables, prod-
uct may be fumigated with natural volatile chemicals. Ethanol vapor treat-
ments retarded softening, darkening, and acid loss, while maintaining
acceptable flavor of whole sweet cherry fruit, thus extending their shelf
life (Bai et al., 2011). When ethanol vapors were applied to whole mango
fruit prior to fresh-cut processing, processed product had reduced micro-
bial counts, but inhibition of ripening was inconsistent and sensory panels
detected some off-flavors in treated fruit (Plotto et al., 2006). Fumigation
of strawberry fruit with phenylethyl alcohol reduced decay and maintained
fruit quality, including fresh-picked flavor (Mo and Sung, 2007). Apple slices
exposed to 10.3 mol/L hexanal prevented decay, and the slices were able
to convert hexanal into esters that contribute to apple flavor (Song et al.,
1996). The potential to supply these volatile treatments in package pro-
duce has recently been demonstrated. Trans-2-hexenal, another naturally
occurring plant volatile with antimicrobial capacity, was encapsulated into
β-cyclodextrins, incorporated into PLA packaging material, and shown to
effectively control microorganisms causing food spoilage (Joo et al., 2012).
A variety of volatile essential oils have also been assessed for their
effectiveness in controlling postharvest deterioration of fresh produce.
The use of lemongrass oil in combination with Bacillus amyloliquefaciens
PPCB004, a microbial antagonist, on peach fruit held in a biodegradable
MAP was found to control decay while maintaining acceptable quality and
flavor (Arrebola et al., 2010). However, fumigation of rabbiteye blueberry

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(Vaccinium virgatum Aiton) fruit with cinnamon oil, linalool, p-cymene,

peppermint leaf oil, Sporan (rosemary and wintergreen oils), or Sporatec
(rosemary, clove, and thyme oils) had limited effects on preventing decay
and caused negative impacts on fruit sensory attributes, including sour-
ness, astringency, juiciness, bitterness, and blueberry-like flavor (Mehra
et al., 2013).

18.5  Flavor Enhancement

There is the potential to increase or alter the flavor of fresh fruits and veg-
etables by supplying them volatile flavor compounds or their precursors.
These volatile compounds can be fed to whole or fresh-cut produce by bulk
fumigation, in-package, liquid infusions, or through incorporation into
coatings. Flavor enhancement can occur through diffusion of flavor com-
pound into the commodity or the metabolism of the compound into other
flavor compounds.
The capacity of apple fruit to utilize volatile compounds present
in the storage atmosphere for ester synthesis has been demonstrated to
enhance fruit flavor (Knee and Hatfield, 1981). Apples previously stored
in CA that were exposed to a mixture of volatile alcohols, aldehydes, and
carboxylic acids had an enhanced aroma compared to untreated fruit
(Kollmannsberger and Berger, 1992). A mixture of aliphatic alcohols in the
precursor atmosphere resulted in a better-balanced apple aroma than one
or two alcohols alone, and sensory panelists could detect a pearlike note
after exposure to a precursor atmosphere that was not present in untreated
fruit. Atmospheres containing aldehydes and carboxylic acids applied to
‘Golden Delicious’ apples selectively enhanced volatile production; how-
ever, significant organoleptic improvement of the fruit was not observed
(Pooter et al., 1987). Whole apples exposed to hexanal vapor had increased
emission of hexanol, hexyl acetate, hexyl butanoate, and hexyl hexanoate
compared to untreated fruit, but enhanced emission of these compounds
was limited to 4–7 days (Fan et al., 2006). Further development of these
methods could provide new approaches to market fruit and vegetables with
enhanced or novel flavors.

18.6  Conclusions and Future Perspectives

Fresh fruit and vegetable flavor is a complex and dynamic property that
is affected by many pre- and postharvest factors. Ensuring good flavor
is delivered to the consumer is critical for consumer satisfaction and the
development of sustainable markets. Whereas postharvest technologies
are designed to prevent decay and breakdown and maintain the good

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appearance of fresh produce, their effect on flavor is often neglected.

Postharvest technologies can preserve, enhance, or degrade produce flavor.
Changes in flavor can result from metabolic changes in flavor compounds,
diffusional loss of volatile flavor compounds, or gains of compounds from
the storage environment. These changes are driven by the postharvest
environment and stress conditions the product may experience during
postharvest handling. More efforts are needed to assess the impact of new
and existing postharvest technologies on the flavor life of fresh fruits and
vegetables. Opportunities exist to enhance product flavor by use of high-
flavored cultivars, better management of harvest maturity, optimization
of fruit ripening, design of flavor-retaining packages and coating formula-
tions, and the development of new technologies to enhance produce flavor.

References
Abdi, N., McGlasson, W.B., Holford, P., Williams, M., and Mizrahi, Y. 1998.
Responses of climacteric and suppressed-climacteric plums to treat-
ment with propylene and 1-methylcyclopropene. Postharvest Biology
and Technology 14, 29–39.
Ali, A., Ong, M.K., and Forney, C.F. 2014. Effect of ozone pre-conditioning on
quality and antioxidant capacity of papaya fruit during ambient storage.
Food Chemistry 142, 19–26.
Allende, A., Marín, A., Buendía, B., Tomás-Barberán, F., and Gil, M.I. 2007.
Impact of combined postharvest treatments (UV-C light, gaseous O3,
superatmospheric O2 and high CO2) on health promoting compounds
and shelf-life of strawberries. Postharvest Biology and Technology 46,
201–211.
Almenar, E., Samsudin, H., Auras, R., and Harte, J. 2010. Consumer accep-
tance of fresh blueberries in bio-based packages. Journal of the Science
of Food and Agriculture 90, 1121–1128.
Argenta, L.C., Fan, X., and Mattheis, J.P. 2003. Influence of 1-methylcyclo-
propene on ripening, storage life, and volatile production by d’Anjou cv.
pear fruit. Journal of Agricultural and Food Chemistry 51, 3858–3864.
Arnon, H., Zaitsev, Y., Porat, R., and Poverenov, E. 2014. Effects of carboxy-
methyl cellulose and chitosan bilayer edible coating on postharvest
quality of citrus fruit. Postharvest Biology and Technology 87, 21–26.
Arrebola, E., Sivakumar, D., Bacigalupo, R., and Korsten, L. 2010. Combined
application of antagonist Bacillus amyloliquefaciens and essential oils for
the control of peach postharvest diseases. Crop Protection 29, 369–377.
Bai, J., Baldwin, E.A., Soliva Fortuny, R.C., Mattheis, J.P., Stanley, R., Perera,
C., and Brecht, J.K. 2004. Effect of pretreatment of intact ‘Gala’ apple
with ethanol vapor, heat, or 1-methylcyclopropene on quality and shelf
life of fresh-cut slices. Journal of the American Society for Horticultural
Science 129, 583–593.

608
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Bai, J., Hagenmaier, R.D., and Baldwin, E.A. 2003. Coating selection for
‘Delicious’ and other apples. Postharvest Biology and Technology 28,
381–390.
Bai, J., Plotto, A., Spotts, R., and Rattanapanone, N. 2011. Ethanol vapor and
saprophytic yeast treatments reduce decay and maintain quality of
intact and fresh-cut sweet cherries. Postharvest Biology and Technology
62, 204–212.
Baigrie, B. 2003. Taints and Off-Flavours in Food. Woodhead Publishing,
Cambridge, England, 203.
Baldwin, E.A. 1994. Edible coatings for fresh fruits and vegetables: Past,
present and future. In Edible Coatings and Films to Improve Food
Quality, Krochta, J., Baldwin, E.A., and Nisperos-Carriedo, M.O., eds.
Technomic Publishing, Lancaster, PA, 25–64.
Baldwin, E.A. 2002. Fruit flavor, volatile metabolism and consumer percep-
tions. In Fruit Quality and Its Biological Basis, Knee, M., ed. CRC Press,
Boca Raton, FL, 89–106.
Baldwin, E.A., Burns, J.K., Kazokas, W., Brecht, J.K., Hagenmaier, R.D.,
Bender, R.J., and Pesis, E. 1999. Effect of two edible coatings with dif-
ferent permeability characteristics on mango (Mangifera indica L.) rip-
ening during storage. Postharvest Biology and Technology 17, 215–226.
Baldwin, E.A., Nisperos-Carriedo, M., Shaw, P.E., and Burns, J.K. 1995. Effect
of coatings and prolonged storage conditions on fresh orange flavor vol-
atiles, degrees brix, and ascorbic acid levels. Journal of Agricultural and
Food Chemistry 43, 1321–1331.
Baldwin, E.A., Plotto, A., and Goodner, K. 2007. Shelf-life versus flavour-life
for fruits and vegetables: How to evaluate this complex trait. Stewart
Postharvest Review 3(1).
Baldwin, E.A., Scott, J.W., Einstein, M.A., Malundo, T.M.M., Carr, B.T.,
Shewfelt, R.L., and Tandon, K.S. 1998. Relationship between sensory
and instrumental analysis for tomato flavor. Journal of the American
Society for Horticultural Science 123, 906–915.
Baldwin, E.A., Scott, J.W., Shewmaker, C.K., and Schuch, W. 2000. Flavor
trivia and tomato aroma: Biochemistry and possible mechanisms for
control of important aroma components. HortScience 35, 1013–1022.
Bangerth, F. 1973. Zur wirkung eines reduzierten drucks, auf physiolo-
gie, qualitat und lagerfahigkeit von obst, gemuse und schnittblumen.
Gartenbauwissenschaft 38, 479–508.
Barry-Ryan, C., and O’Beirne, D. 1998. Quality and shelf-life of fresh cut
carrot slices as affected by slicing method. Journal of Food Science 63,
851–856.
Beaulieu, J.C. 2006. Volatile changes in cantaloupe during growth, matura-
tion, and in stored fresh-cuts prepared from fruit harvested at various
maturities. Journal of the American Society for Horticultural Science 131,
127–139.
Beaulieu, J.C. 2007. Effect of UV irradiation on cut cantaloupe: Terpenoids
and esters. Journal of Food Science 72, S272–S281.

609
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Beaulieu, J.C., and Baldwin, E. 2002. Flavor and aroma of fresh-cut fruits and
vegetables. In Fresh-Cut Fruits and Vegetables: Science, Technology and
Market, Lamikanra, O., ed. CRC Press, Boca Raton, FL, 391–425.
Beaulieu, J.C., Ingram, D.A., Lea, J.M., and Bett-Garber, K.L. 2004. Effect of
harvest maturity on the sensory characteristics of fresh-cut cantaloupe.
Journal of Food Science 69, S250–S258.
Bender, R.J., Brecht, J.K., Sargent, S.A., and Huber, D.J. 2000. Mango tol-
erance to reduced oxygen levels in controlled atmosphere storage.
Journal of the American Society for Horticultural Science 125, 707–713.
Besada, C., Sanchez, G., Salvador, A., and Granell, A. 2013. Volatile com-
pounds associated to the loss of astringency in persimmon fruit revealed
by untargeted GC-MS analysis. Metabolomics 9, 157–172.
Bett, K.L., Ingram, D.A., Grimm, C.C., Lloyd, S.W., Spanier, A.M., Miller,
J.M., Gross, K.C., Baldwin, E.A., and Vinyard, B.T. 2001. Flavor of fresh-
cut Gala apples in barrier film packaging as affected by storage time.
Journal of Food Quality 24, 141–156.
Blankenship, S.M., and Dole, J.M. 2003. 1-Methylcyclopropene: A review.
Postharvest Biology and Technology 28, 1–25.
Bramlage, W.J., and Meir, S. 1990. Chilling injury of crops of temperate ori-
gin. In Chilling Injury of Horticultural Crops, Wang, C.Y., ed. CRC Press,
Boca Raton, FL, 37–49.
Brody, A.L. 2002. Flavor scalping: Quality loss due to packaging. Food
Technology 56, 124.
Caleb, O.J., Opara, U.L., Mahajan, P.V., Manley, M., Mokwena, L., and
Tredoux, A.G.J. 2013. Effect of modified atmosphere packaging and
storage temperature on volatile composition and postharvest life of
minimally-processed pomegranate arils (cvs. ‘Acco’ and ‘Herskawitz’).
Postharvest Biology and Technology 79, 54–61.
Cameron, A.C., Talasila, P.C., and Joles, D.W. 1995. Predicting film permeabil-
ity needs for modified-atmosphere packaging of lightly processed fruits
and vegetables. HortScience 30, 25–34.
Cliff, M.A., Lau, O.L., and King, M.C. 1998. Sensory characteristics of con-
trolled atmosphere- and air-stored ‘Gala’ apples. Journal of Food Quality
21, 239–249.
Cliff, M.A., Toivonen, P.M.A., Forney, C.F., Liu, P., and Lu, C. 2010. Quality of
fresh-cut apple slices stored in solid and micro-perforated film packages
having contrasting O2 headspace atmospheres. Postharvest Biology and
Technology 58, 254–261.
Corollaro, M.L., Endrizzi, I., Bertolini, A., Aprea, E., Demattè, M.L.,
Costa, F., Biasioli, F., and Gasperi, F. 2013. Sensory profiling of apple:
Methodological aspects, cultivar characterisation and postharvest
changes. Postharvest Biology and Technology 77, 111–120.
Crisosto, C.H. 1999. Optimum procedures for ripening stone fruit. In
Management of Fruit Ripening. Postharvest Horticulture Series 9.
University of California, Davis, 28–30.

610
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Crisosto, C.H., Garner, D., and Crisosto, G. 2002a. Carbon dioxide-enriched

atmospheres during cold storage limit losses from Botrytis but acceler-
ate rachis browning of ‘Redglobe’ table grapes. Postharvest Biology and
Technology 26, 181–189.
Crisosto, C.H., Garner, D., and Crisosto, G. 2002b. High carbon dioxide atmo-
spheres affect stored ‘Thompson Seedless’ table grapes. HortScience
37, 1074–1078.
Daillant-Spinnler, B., MacFie, H.J.H., Beyts, P.K., and Hedderley, D. 1996.
Relationships between perceived sensory properties and major prefer-
ence directions of 12 varieties of apples from the southern hemisphere.
Food Quality and Preference 7, 113–126.
Daniels-Lake, B.J., Prange, R.K., Gaul, S.O., McRae, K.B., de Antueno, R.,
and McLachlan, D. 2007. A musty “off” flavor in Nova Scotia potatoes
is associated with 2,4,6-trichloroanisole released from pesticide-treated
soils and high soil temperature. Journal of the American Society for
Horticultural Science 132, 112–119.
de Kock, A., and Taylor, M. 2014. Cold storage of plums for the South
African market, p. 6. http://www.experico.co.za/ (Accessed January
14, 2014).
Dixon, J., and Hewett, E.W. 2000. Factors affecting apple aroma/flavour
volatile concentration: A review. New Zealand Journal of Crop and
Horticultural Sciences 28, 155–173.
Drewnowski, A. 1997. Taste preferences and food intake. Annual Review of
Nutrition 17, 237–253.
Duvenhage, A.J., Jooste, M., and Johnson, S.A. 2012. Irradiation as a posthar-
vest quarantine treatment for a new pluot cultivar. Scientia Horticulturae
147, 64–70.
Eccher Zerbini, P. 2008. Role of maturity for improved flavour. In Fruit and
Vegetable Flavour: Recent Advances and Future Prospects, Brückner, B.,
and Wyllie, S.G., eds. CRC Press, Boca Raton, FL, 150–164.
Echeverría, G., Lara, I., Fuentes, T., López, M.L., Graell, J., and Puy, J. 2004.
Assessment of relationships between sensory and instrumental quality
of controlled-atmosphere-stored ‘Fuji’ apples by multivariate analysis.
Journal of Food Science 69, S368–S375.
Fallik, E., Archbold, D.D., Hamilton-Kemp, T.R., Loughrin, J.H., and Collins,
R.W. 1997. Heat treatment temporarily inhibits aroma volatile com-
pound emission from Golden Delicious apples. Journal of Agricultural
and Food Chemistry 45, 4038–4041.
Fan, L., Song, J., Beaudry, R.M., and Hildebrand, P.D. 2006. Effect of hexanal
vapor on spore viability of Penicillin expansum, lesion development on
whole apples and fruit volatile biosynthesis. Journal of Food Science 71,
M105–M109.
Fan, L., Song, J., Forney, C.F., and Jordan, M.A. 2005a. Ethanol production
and chlorophyll fluorescence in heat-stressed apple fruit during ­storage.
Acta Horticulturae 682, 901–907.

611
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Fan, L., Song, J., Forney, C.F., and Jordan, M.A. 2005b. Ethanol production and
chlorophyll fluorescence predict breakdown of heat-stressed apple fruit
during cold storage. Journal of the American Society for Horticultural
Science 130, 237–243.
Fan, L., Song, J., Forney, C.F., and Jordan, M.A. 2011. Fruit maturity affects
the response of apples to heat stress. Postharvest Biology and Technology
62, 35–42.
Fan, X., and Mattheis, J.P. 1999. Impact of 1-methylcyclopropene and methyl
jasmonate on apple volatile production. Journal of Agricultural and Food
Chemistry 47, 2847–2853.
Fan, X., and Mattheis, J.P. 2000. Reduction of ethylene-induced physiologi-
cal disorders of carrots and iceberg lettuce by 1-methylcyclopropene.
HortScience 35, 1312–1314.
Fellman, J.K., Miller, T.W., Mattinson, D.S., and Mattheis, J.P. 2000. Factors
that influence biosynthesis of volatile flavor compounds in apple fruits.
HortScience 35, 1026–1033.
Fidler, J.C., and North, C.J. 1971. The effect of periods of anaerobiosis on the
storage of apples. Journal of Horticultural Science 46, 213–221.
Forney, C.F. 1995. Hot-water dips extend the shelf life of fresh broccoli.
HortScience 30, 1054–1057.
Forney, C.F. 2003. Postharvest response of horticultural products to ozone.
In Postharvest Oxidative Stress in Horticultural Crops, Hodges, D.M., ed.
Food Products Press, New York, 13–54.
Forney, C.F. 2007. New innovations in the packaging of fresh-cut produce.
Acta Horticulturae 746, 53–60.
Forney, C.F. 2008. Flavour loss during postharvest handling and marketing of
fresh-cut produce. Stewart Postharvest Review 5(3), 1–10.
Forney, C.F., Fan, L., Song, J., and Jordan, M.A. 2011. Heat stress and fruit
maturity affects the volatile composition of apples during storage.
HortScience 46, S271.
Forney, C.F., and Jordan, M.A. 1995. Methanethiol production by Brassica
vegetables held in anaerobic atmospheres. HortScience 30, 823.
Forney, C.F., and Jordan, M.A. 1998. Induction of volatile compounds in broc-
coli by postharvest hot-water dips. Journal of Agricultural and Food
Chemistry 46, 5295–5301.
Forney, C.F., and Jordan, M.A. 1999. Anaerobic production of methanethiol
and other compounds by Brassica vegetables. HortScience 34, 696–699.
Forney, C.F., Jordan, M.A., Fan, L., Doucette, C., and Grant, R. 2012a.
Influence of packaging on loss of aroma and quality of diced red onions.
Acta Horticulturae 934, 523–530.
Forney, C.F., Jordan, M.A., and Nicholas, K.U.K.G. 2000a. Volatile emissions
and chlorophyll fluorescence as indicators of freezing injury in apple
fruit. HortScience 35, 1283–1287.
Forney, C.F., Kalt, W., and Jordan, M.A. 2000b. The composition of strawberry
aroma is influenced by cultivar, maturity, and storage. HortScience 35,
1022–1026.

612
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Forney, C.F., Kalt, W., Jordan, M.A., Vinqvist-Tymchuk, M.R., and Fillmore,
S.A.E. 2012b. Blueberry and cranberry fruit composition during devel-
opment. Journal of Berry Research 2, 169–177.
Forney, C.F., Mattheis, J.P., and Austin, R.K. 1991. Volatile compounds pro-
duced by broccoli under anaerobic conditions. Journal of Agricultural
and Food Chemistry 39, 2257–2259.
Forney, C.F., Mattheis, J.P., and Baldwin, E.A. 2009. Effects on flavor. In
Modified and Controlled Atmospheres for the Storage, Transportation,
and Packaging of Horticultural Commodities, Yahia, E.M., ed. CRC
Press, Boca Raton, FL, 119–158.
Forney, C.F., Song, J., Hildebrand, P.D., Fan, L., and McRae, K.B. 2007.
Interactive effects of ozone and 1-methylcyclopropene on decay resis-
tance and quality of stored carrots. Postharvest Biology and Technology
45, 341–348.
Forney, C.F., and Yaganza, E.S. 2011. Fresh-cut produce and the use of biaxial
stretched films. In Biaxial Stretching of Film Principles and Applications,
DeMeuse, M.T., ed. Woodhead Publishing, Oxford, 143–164.
Golding, J.B., Shearer, D., Wyllie, S.G., and McGlasson, W.B. 1998. Application
of 1-MCP and propylene to identify ethylene-dependent ripening pro-
cesses in mature banana fruit. Postharvest Biology and Technology 14,
87–98.
Gorny, J.R. 2005. Leveraging innovative fresh-cut technologies for competi-
tive advantage. Acta Horticulturae 687, 141–147.
Hagenmaier, R.D., and Shaw, P.E. 2002. Changes in volatile components of
stored tangerines and other specialty citrus fruits with different coat-
ings. Journal of Food Science 67, 1742–1745.
Hardenburg, R.E., Watada, A.E., and Wang, C.Y. 1986. The Commercial Storage
of Fruits, Vegetables, and Florist and Nursery Stocks. USDA Agricultural
Handbook 66. U.S. Department of Agriculture, Washington, DC, 130.
Hoa, T.T., Ducamp, M.N., Lebrun, M., and Baldwin, E.A. 2002. Effect of dif-
ferent coating treatments on the quality of mango fruit. Journal of Food
Quality 25, 471–486.
Holley, A. 2006. Processing information about flavour. In Flavour in Food,
Voilley, A., and Etiévant, P., eds. CRC Press, Boca Raton, FL, 36–61.
Iyer, M.M., Sacks, G.L., and Padilla-Zakour, O.I. 2010. Impact of h ­ arvesting
and processing conditions on green leaf volatile development
and ­phenolics in concord grape juice. Journal of Food Science 75,
C297–C304.
Järvenpää, E.P., Zhang, Z., Huopalahti, R., and King, J.W. 1998a. Determination
of fresh onion (Allium cepa L.) volatiles by solid phase microextraction
combined with gas chromatography-mass spectrometry. European
Food Research and Technology 207, 39–43.
Joo, M.J., Merkel, C., Auras, R., and Almenar, E. 2012. Development and
characterization of antimicrobial poly(l-lactic acid) containing trans-
2-­
hexenal trapped in cyclodextrins. International Journal of Food
Microbiology 153, 297–305.

613
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Kader, A.A., Morris, L.L., Stevens, M.A., and Albright-Holton, M. 1978.

Composition and flavor quality of fresh market tomatoes as influenced
by some postharvest handling procedures. Journal of the American
Society for Horticultural Science 103, 6–13.
Ke, D., Zhou, L., and Kader, A.A. 1994. Mode of oxygen and carbon dioxide
action on strawberry ester biosynthesis. Journal of the American Society
for Horticultural Science 119, 971–975.
Ke, D.Y., Goldstein, L., O’Mahony, M., and Kader, A.A. 1991. Effects of short-
term exposure of low O2 and high CO2 atmospheres on quality attri-
butes of strawberries. Journal of Food Science 56, 50–54.
Klein, J.D., and Lurie, S. 1992. Prestorage heating of apple fruit for enhanced
postharvest quality: Interaction of time and temperature. HortScience
27, 326–328.
Knee, M., and Hatfield, S.G.S. 1981. The metabolism of alcohols by apple fruit
tissue. Journal of the Science of Food and Agriculture 32, 593–600.
Kollmannsberger, H., and Berger, R.G. 1992. Precursor atmosphere stor-
age induced flavour changes in apples cv. Red Delicious. Chemie,
Mikrobiologie, Technologie der Lebensmittel 14, 81–86.
Lalel, H.J.D., Singh, Z., and Tan, S.C. 2003. Maturity stage at harvest affects
fruit ripening, quality and biosynthesis of aroma volatile compounds
in ‘Kensington Pride’ mango. Journal of Horticultural Science and
Biotechnology 78, 225–233.
Lamikanra, O., Kuenenam, D., Ukuku, D., and Bett-Garber, K.L. 2005. Effect
of processing under ultraviolet light on the shelf life of fresh-cut canta-
loupe melon. Journal of Food Science 70, 534–539.
Lamikanra, O., and Richard, O.A. 2002. Effect of storage on some volatile
aroma compounds in fresh-cut cantaloupe melon. Journal of Agricultural
and Food Chemistry 50, 4043–4047.
Lamikanra, O., and Richard, O.A. 2004. Storage and ultraviolet-induced tissue
stress effects on fresh-cut pineapple. Journal of the Science of Food and
Agriculture 84, 1812–1816.
Lamikanra, O., and Watson, M.A. 2003. Temperature and storage duration
effects on esterase activity in fresh-cut cantaloupe melon. Journal of
Food Science 68, 790–793.
Larsen, M., and Watkins, C.B. 1995. Firmness and concentrations of acetal-
dehyde, ethyl acetate and ethanol in strawberries stored in controlled
and modified atmospheres. Postharvest Biology and Technology 5,
39–50.
Lèlievre, J.M., Latché, A., Jones, B., Bouzayen, M., and Pech, J.C. 1997.
Ethylene and fruit ripening. Physiologia Plantarum 101, 727–739.
Loayza, F.E., Brecht, J.K., Plotto, A., Baldwin, E.A., and Bai, J. 2012. Evaluation
of the impact of hot water treatment on the sensory quality of fresh
tomatoes. Acta Horticulturae 934, 1305–1312.
López, M.L., Miró, R., and Graell, J. 2001. Quality and aroma production of
Doyenne du Comice pears in relation to harvest date and storage atmo-
sphere. Food Science and Technology International 7, 493–500.

614
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Lubbers, S. 2006. Texture-aroma interactions. In Flavour in Food, Voilley, A., and

Etiévant, P., eds. Woodhead Publishing, Cambridge, England, 327–344.
Lurie, S. 1998. Postharvest heat treatments. Postharvest Biology and
Technology 14, 257–269.
Lurie, S., and Crisosto, C.H. 2005. Chilling injury in peach and nectarine.
Postharvest Biology and Technology 37, 195–208.
Lurie, S., Pre-Aymard, C., Ravid, U., Larkov, O., and Fallik, E. 2002. Effect
of 1-methylcyclopropene on volatile emission and aroma in cv. Anna
apples. Journal of Agricultural and Food Chemistry 50, 4251–4256.
Maria, A.M.L., Vergara, M., Bravo, C., Pereira, M., and Moggia, C. 2007.
Development of aroma compounds and sensory quality of ‘Royal Gala’
apples during storage. Journal of Horticultural Science and Biotechnology
82, 403–413.
Martínez-Romero, D., Alburquerque, N., Valverde, J.M., Guillén, F., Castillo,
S., Valero, D., and Serrano, M. 2006. Postharvest sweet cherry quality
and safety maintenance by Aloe vera treatment: A new edible coating.
Postharvest Biology and Technology 39, 93–100.
Mattheis, J.P. 1991. Changes in headspace volatiles during physiological
development of Bisbee Delicious apple fruit. Journal of Agricultural and
Food Chemistry 39, 1902–1906.
Mattheis, J.P., Buchanan, D.A., and Fellman, J.K. 1995. Volatile compound
production by Bisbee Delicious apples after sequential atmosphere stor-
age. Journal of Agricultural and Food Chemistry 43, 194–199.
Mattheis, J.P., Fan, X., and Argenta, L.C. 2005. Interactive responses of Gala
apple fruit volatile production to controlled atmosphere storage and
chemical inhibition of ethylene action. Journal of Agricultural and Food
Chemistry 53, 4510–4516.
Maul, F., Sargent, S.A., Sims, C.A., Baldwin, E.A., Balaban, M.O., and Huber,
D.J. 2000. Tomato flavor and aroma quality as affected by storage tem-
perature. Journal of Food Science 65, 1228–1237.
Mehra, L.K., MacLean, D.D., Shewfelt, R.L., Smith, K.C., and Scherm, H.
2013. Effect of postharvest biofumigation on fungal decay, sensory
quality, and antioxidant levels of blueberry fruit. Postharvest Biology and
Technology 85, 109–115.
Meilgaard, M., Civille, G.V., and Carr, B.T. 1999. Sensory Evaluation
Techniques. 3rd ed. CRC Press, Boca Raton, FL.
Misry, B.S., Reineccius, T., and Olson, L.K. 1997. Gas chromotography-­
olfactometry for the determination of key odorants in foods. In
Techniques for Analyzing Food Aroma, Marsili, R., ed. Marcel Dekker,
New York, 265–292.
Mitcham, E.J., and McDonald, R.E. 1993. Respiration rate, internal atmo-
sphere, and ethanol and acetaldehyde accumulation in heat-treated
mango fruit. Postharvest Biology and Technology 3, 77–86.
Mo, E.K., and Sung, C.K. 2007. Phenylethyl alcohol (PEA) application slows
fungal growth and maintains aroma in strawberry. Postharvest Biology
and Technology 45, 234–239.

615
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Myung, K., Hamilton-Kemp, T.R., and Archbold, D.D. 2006. Biosynthesis of

trans-2-hexenal in response to wounding in strawberry fruit. Journal of
Agricultural and Food Chemistry 54, 1442–1448.
Norton, J.S., Charig, A.J., and Demoranville, I.E. 1968. The effect of ozone
on storage of cranberries. Proceedings of the American Society for
Horticultural Sciences 93, 792–796.
Obenland, D., Collin, S., Sievert, J., and Arpaia, M.L. 2013. Mandarin flavor
and aroma volatile composition are strongly influenced by holding tem-
perature. Postharvest Biology and Technology 82, 6–14.
Obenland, D.M., Arpaia, M.L., Austin, R.K., and MacKey, B.E. 1999. High-
temperature forced-air treatment alters the quantity of flavor-related,
volatile constituents present in navel and valencia oranges. Journal of
Agricultural and Food Chemistry 47, 5184–5188.
Pariasca, J.A.T., Miyazaki, T., Hisaka, H., Nakagawa, H., and Sato, T. 2001.
Effect of modified atmosphere packaging (MAP) and controlled
atmosphere (CA) storage on the quality of snow pea pods (Pisum
­
sativum L. var. saccharatum). Postharvest Biology and Technology 21,
213–223.
Parsons, C.S., Anderson, R.E., and Penny, R.W. 1970. Storage of mature-green
tomatoes in controlled atmospheres. Journal of the American Society for
Horticultural Science 95, 791–794.
Paull, R.E. 1990. Chilling injury of crops of tropical and subtropical origin. In
Chilling Injury of Horticultural Crops, Wang, C.Y., ed. CRC Press, Boca
Raton, FL, 17–36.
Pelayo, C., Ebeler, S.E., and Kader, A.A. 2003. Postharvest life and flavor qual-
ity of three strawberry cultivars kept at 5°C in air or air + 20 kPa CO2.
Postharvest Biology and Technology 27, 171–183.
Pérez, A.G., Sanz, C., Ríos, J.J., Olías, R., and Olías, J.M. 1999. Effects of ozone
treatment on postharvest strawberry quality. Journal of Agricultural and
Food Chemistry 47, 1652–1656.
Pesis, E., Zauberman, G., and Avissar, I. 1991. Induction of certain aroma
volatiles in feijoa fruit by postharvest application of acetaldehyde or
anaerobic conditions. Journal of the Science of Food and Agriculture 54,
329–337.
Plotto, A., Bai, J., Narciso, J.A., Brecht, J.K., and Baldwin, E.A. 2006. Ethanol
vapor prior to processing extends fresh-cut mango storage by decreas-
ing spoilage, but does not always delay ripening. Postharvest Biology and
Technology 39, 134–145.
Plotto, A., Goodner, K.L., Baldwin, E.A., Bai, J., and Rattanapanone, N. 2004.
Effect of polysaccharide coatings on quality of fresh cut mangoes
(Mangifera indica). Proceedings of the Florida State Horticultural Society
117, 381–388.
Plotto, A., McDaniel, M.R., and Mattheis, J.P. 1999. Characterization of ‘Gala’
apple aroma and flavor: Differences between controlled atmosphere
and air storage. Journal of the American Society for Horticultural Science
124, 416–423.

616
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Podoski, B.W., Sims, C.A., Sargent, S.A., Price, J.F., Chandler, C.K., and
Okeefe, S.F. 1997. Effects of cultivar, modified atmosphere, and pre-har-
vest conditions on strawberry quality. Proceedings of the Florida State
Horticulture Society 110, 246–252.
Pooter, H.L.D., Acker, M.R.V., and Schamp, N.M. 1987. Aldehyde metabolism
and the aroma quality of stored Golden Delicious apples. Phytochemistry
26, 89–92.
Portela, S.I., and Cantwell, M.I. 1998. Quality changes of minimally processed
honeydew melons stored in air or controlled atmosphere. Postharvest
Biology and Technology 14, 351–357.
Portela, S.I., and Cantwell, M.I. 2001. Cutting blade sharpness affects appear-
ance and other quality attributes of fresh-cut cantaloupe melon. Journal
of Food Science 66, 1265–1270.
Randle, W.M., and Lancaster, J.E. 2002. Sulphur compounds in alliums in
relation to flavour quality. In Allium Crops Science: Recent Advances,
Rabinowitch, H.D., and Currah, L., eds. CAB International, Wallingford,
UK, 329–356.
Ravanfar, R., Niakousari, M., and Maftoonazad, N. 2012. Postharvest sour
cherry quality and safety maintenance by exposure to hot-water or treat-
ment with fresh Aloe vera gel. Journal of Food Science and Technology,
1–5.
Ridley, J.D., and Sims, E.T. 1967. The response of peaches to ozone dur-
ing storage. Technical Bulletin 1027. South Carolina Agricultural
Experiment Station, Clemson University, Clemson, SC, 24.
Riley, J.C.M., and Thompson, J.E. 1998. Ripening-induced acceleration of
volatile aldehyde generation following tissue disruption in tomato fruit.
Physiologia Plantarum 104, 571–576.
Rocha, A.M.C.N., Brochado, C.M., Kirby, R., and Morais, A.M.M.B. 1995.
Shelf-life of chilled cut orange determined by sensory quality. Food
Control 6, 317–322.
Rupérez, P., and Toledano, G. 2003. Celery by-products as a source of man-
nitol. European Food Research and Technology 216, 224–226.
Sabir, F.K., and Sabir, A. 2013. Quality response of table grapes (Vitis
­vinifera L.) during cold storage to postharvest cap stem excision and hot
water treatments. International Journal of Food Science and Technology
48, 999–1006.
Saftner, R.A., Abbott, J.A., Conway, W.S., Barden, C.L., and Vinyard, B.T.
2002. Instrumental and sensory quality characteristics of ‘Gala’ apples
in response to prestorage heat, controlled atmosphere, and air storage.
Journal of the American Society for Horticultural Science 127, 1006–1012.
Sajilata, M.G., Savitha, K., Singhal, R.S., and Kanetkar, V.R. 2007. Scalping of
flavors in packaged foods. Comprehensive Reviews in Food Science and
Food Safety 6, 17–35.
Salle, C. 2006. Odour-taste interactions in flavour perception. In Flavour in
Food, Voilley, A., and Etiévant, P., eds. Woodhead Publishing, Cambridge,
England, 345–368.

617
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Saltveit, M.E. 2004. Respiratory metabolism. In The Commercial Storage of

Fruits, Vegetables, and Florist and Nursery Stocks, Gross, K.C., Wang,
C.Y., and Saltveit, M.E., eds. USDA Handbook 66. Available from http://
www.ba.ars.usda.gov/hb66/contents.html (accessed August 16, 2010).
Saltveit, M.E., and Morris, L.L. 1990. Overview on chilling injury of horti-
cultural crops. In Chilling Injury of Horticultural Crops, Wang, C.Y., ed.
CRC Press, Boca Raton, FL, 3–15.
Sanz, C., Olías, J.M., and Pérez, A.G. 1997. Aroma biochemistry of fruits and
vegetables. In Phytochemistry of Fruits and Vegetables, Tomás-Barberán,
F.A., and Robins, R.J., eds. Clarendon Press, Oxford, 125–155.
Schomer, H.A., and McColloch, L.P. 1948. Ozone in relation to storage of apples.
USDA Circular 765. U.S. Department of Agriculture, Washington, DC.
Serra Bonvehí, J., and Ventura Coll, F. 1997. Evaluation of bitterness and astrin-
gency of polyphenolic compounds in cocoa powder. Food Chemistry 60,
365–370.
Shaw, P.E., Moshonas, M.G., Nisperos-Carriedo, M.O., and Carter, R.D.
1992. Controlled-atmosphere treatment of freshly harvested oranges at
elevated temperature to increase volatile flavor components. Journal of
Agricultural and Food Chemistry 40, 1041–1045.
Silva, J.M., Correia, L.C.S.A., Moura, N.P., Salgado, P.L., Maciel, M.I.S., and
Villar, H.P. 2009. Sensorial analysis of strawberry submitted to the tech-
nology of ionising radiation. Acta Horticulturae 842, 863–866.
Smyth, A.B., Song, J., and Cameron, A.C. 1998. Modified atmosphere packaged
cut iceberg lettuce: Effect of temperature and O2 partial pressure on respira-
tion and quality. Journal of Agricultural and Food Chemistry 46, 4556–4562.
Song, J., Fan, L., Forney, C.F., Hildebrand, P.D., Jordan, M.A., Renderos, W.,
and McRae, K.B. 2003. Ozone and 1-MCP treatments affect the quality
and storage life of fresh carrots. Acta Horticulturae 628, 295–301.
Song, J., Fan, L., Forney, C.F., and Jordan, M.A. 2001. Using volatile emis-
sions and chlorophyll fluorescence as indicators of heat injury in apples.
Journal of the American Society of Horticultural Science 126, 771–777.
Song, J., and Forney, C.F. 2008. Flavour volatile production and regulation in
fruit. Canadian Journal of Plant Science 88, 537–550.
Song, J., Leepipattanawit, R., Deng, W., and Beaudry, R.M. 1996. Hexanal
vapor is a natural, metabolizable fungicide: Inhibition of fungal activity
and enhancement of aroma biosynthesis in apple slices. Journal of the
American Society for Horticultural Science 121, 937–942.
Streif, J., and Bangerth, F. 1988. Production of volatile aroma substances by
‘Golden Delicious’ apple fruits after storage for various times in different
CO2 and O2 concentrations. Journal of Horticultural Science 63, 193–199.
Sugar, D. and Basile, S.R. 2009. Low-temperature induction of ripening
capacity in ‘Comice’ and ‘Bosc’ pears as influenced by fruit maturity.
Postharvest Biol. Technol. 51, 278–280.

618
 IM P A C T O F P OST H AR V EST TE C H NOLOGIES

Tandon, K.S., Baldwin, E.A., and Shewfelt, R.L. 2000. Aroma perception
of ­ individual volatile compounds in fresh tomatoes (Lycopersicon
­esculentum Mill.) as affected by the medium of evaluation. Postharvest
Biology and Technology 20, 261–268.
Taylor, A.J., and Linforth, R.S.T. 1994. Methodology for measuring volatile
profiles in the mouth and nose during eating. In Trends in Flavour
Research, Maarse, J., and Heij, V.D., eds. Elsevier Science, New York,
3–14.
Teles, C.S., Benedetti, B.C., Gubler, W.D., and Crisosto, C.H. 2014. Prestorage
application of high carbon dioxide combined with controlled atmosphere
storage as a dual approach to control Botrytis cinerea in organic ‘Flame
Seedless’ and ‘Crimson Seedless’ table grapes. Postharvest Biology and
Technology 89, 32–39.
Thomas, M., and Fidler, J.C. 1933. Studies to zymasis. IV. Zymasis by apples
in relation to oxygen concentration. Biochemistry Journal 27, 1629–1642.
Tietel, Z., Lewinsohn, E., Fallik, E., and Porat, R. 2012. Importance of storage
temperatures in maintaining flavor and quality of mandarins. Postharvest
Biology and Technology 64, 175–182.
Tindale, C.R., Whitfield, F.B., Levingston, S.D., and Nguyen, T.H.L. 1989.
Fungi isolated from packaging materials: Their role in the production
of 2,4,6-trichloroanisole. Journal of the Science of Food and Agriculture
49, 437–447.
Toye, J.D., Glucina, P.G., and Minamide, T. 1987. Removal of astringency and
storage of ‘Hiratanenashi’ persimmon fruits. New Zealand Journal of
Experimental Agriculture 15, 351–355.
Tudela, J.A., Marín, A., Martínez-Sánchez, A., Luna, M.C., and Gil, M.I. 2013.
Preharvest and postharvest factors related to off-odours of fresh-cut ice-
berg lettuce. Postharvest Biology and Technology 86, 463–471.
Tzortzakis, N., Borland, A., Singleton, I., and Barnes, J. 2007. Impact of atmo-
spheric ozone-enrichment on quality-related attributes of tomato fruit.
Postharvest Biology and Technology 45, 317–325.
Usenik, V., Fabcic, J., and Stampar, F. 2008. Sugars, organic acids, phenolic
composition and antioxidant activity of sweet cherry (Prunus avium L.).
Food Chemistry 107, 185–192.
Vanoli, M., Visai, C., and Rizzolo, A. 1995. The influence of harvest date on the
volatile composition of ‘Starkspur Golden’ apples. Postharvest Biology
and Technology 6, 225–234.
Voilley, A., and Souchon, I. 2006. Flavour retention and release from the food
matrix: An overview. In Flavour in Food, Voilley, A., and Etiévant, P., eds.
Woodhead Publishing, Cambridge, England, 117–132.
Voirol, E., and Daget, N. 1987. Nasal and retronasal olfactory perception of
a meat aroma. In Flavor Science and Technology, Martens, M., Dalen,
G.A., and Russwurm, H., eds. Wiley, New York, 309–316.

619
P OST H AR V EST RI P ENING P H Y SIOLOG Y

Wang, C.Y. 2004. Chilling and freezing injury. In The Commercial Storage of
Fruits, Vegetables, and Florist and Nursery Stocks, Gross, K.C., Wang,
C.Y., and Saltveit, M.E., eds. USDA Handbook 66. Available from http://
www.ba.ars.usda.gov/hb66/contents.html (accessed August 16, 2010).
Watada, A.E., and Aulenbach, B.B. 1979. Chemical and sensory qualities of
fresh market tomatoes. Journal of Food Science 44, 1013–1016.
Watkins, C.B. 2006. The use of 1-methylcyclopropene (1-MCP) on fruits and
vegetables. Biotechnology Advances 24, 389–409.
Whitaker, B.D. 2008. Postharvest flavor deployment and degradation in fruits
and vegetables. In Fruit and Vegetable Flavour: Recent Advances and
Future Prospects, Brücker, B., and Wyllie, S.G., eds. CRC Press, Boca
Raton, FL, 103–131.
Xi, W.P., Zhang, B., Liang, L., Shen, J.Y., Wei, W.W., Xu, C.J., Allan, A.C.,
Ferguson, I.B., and Chen, K.S. 2012. Postharvest temperature influ-
ences volatile lactone production via regulation of acyl-CoA oxidases in
peach fruit. Plant, Cell and Environment 35, 534–545.
Yahia, E.M., Liu, F.W., and Acree, T.E. 1990. Changes in some odor-active
volatiles in controlled atmosphere stored apples. Journal of Food Quality
13, 185–202.
Yahia, E.M., Rivera, M., and Hernandez, O. 1992. Responses of papaya to
short-term insecticidal oxygen atmosphere. Journal of the American
Society for Horticultural Science 117, 96–99.

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FRUIT & VEGETABLE PRODUCTS

Postharvest Ripening
Physiology of Crops
Postharvest Ripening Physiology of Crops is a comprehensive interdisciplinary
reference source for the various aspects of fruit ripening and postharvest be-
havior. It focuses on the postharvest physiology, biochemistry, and molecular
biology of ripening and provides an overview of fruits and vegetables, including
chapters on the postharvest quality of ornamental plants and molecular biology
of flower senescence.

It describes various developments that have taken place in the last decade
with respect to identifying and altering the function of ripening-related genes.
Taking clues from studies in grape and tomato as model fruits, the book reviews
a few case studies and gives you a detailed account of molecular regulation of
fruit ripening, and signal transduction and internal atmospheres in relation to
fruit ripening. It also presents an overview of methods utilized in fruit proteomics,
as well as a global proteome and systems biology analysis of fruits during rip-
ening, and discusses the basics of dormancy, its molecular and physiological
basis, and methods to break the dormancy.

The book provides an overview of the most important metabolic pathways

and genes that control volatile biosynthesis in model fruits, including tropical,
subtropical, and temperate fruits, with a special emphasis on fruit ripening and
the role of ethylene during this process. It presents a brief description of the
composition of volatiles in various fruit species and addresses the influences of
preharvest factors and postharvest technologies on fruit aroma, basic mech-
anisms responsible for postharvest flavor change in fresh produce, and the
potential impacts of various postharvest technologies on flavor.

K24683
ISBN: 978-1-4987-0380-2
90000

9 781498 703802