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"
DEDICATION
To JEHOVAH God Almighty be all the Glory
This work is dedicated to my guardians and family for their immeasurable
support, inspiration, guidance, love and prayers.
ACKNOWLEDGEMENT
I am grateful, first and foremost to Jehovah God Almighty, for His Divine
Grace, favour and sustenance to complete this work. I would like to express
my deep gratitude and appreciation to my supervisors DR. HM AMOATEY,
Head of Department of Nuclear Agriculture and Radiation Processing and
PROF. GYP KLU, former Head of Department of Nuclear Agriculture and
Radiation Processing of the Graduate School of Nuclear and Allied Sciences,
University of Ghana (Atomic campus) for their valuable contributions,
guidance, patience, encouragement and constructive suggestions towards the
success of this work.
"
"
the
Laboratory
of
Molecular Systematics,
Conservation of Bryophytes,
Phylogeography
and
"
"
TABLE OF CONTENT
CHAPTER ONE0..1
INTRODUCTION........1
1.1 GENERAL INTRODUCTION0......1
1.1.1 PROBLEM STATEMENT......02
1.1.2 JUSTIFICATION AND RELEVANCE OF THE STUDY03
1.1.3 OBJECTIVES OF THE RESEARCH.06
REFERENCES0..007
CHAPTER TWO......8
LITERATURE REVIEW.8
2.1 ORIGIN, DISTRIBUTION AND UTILISATION0008
2.2 TAXONOMIC CLASSIFICATION AND BOTANY0010
2.3 GENETICS AND CYTOLOGY...11
2.4 SOIL, CLIMATIC AND AGRONOMIC REQUIREMENTS......14
2.5 GROWTH AND DEVELOPMENT..00014
2.6 POLLINATION AND FERTILISATION..0016
2.7 RATOONING..017
2.8 NUTRIENT CONTENTS AND HEALTH BENEFITS...18
2.8.1 Biochemical and nutritional composition18
2.8.2 Nutritional healing properties0019
2.8.3 Bioactive properties019
"
"
CHAPTER THREE...057
MORPHOLOGICAL CHARACTERISATION OF 29 ACCESSIONS OF
OKRA (Abelmoschus spp L.)..00057
3.1 INTRODUCTION00057
3.1.1 Objectives of the Study58
3.2 MATERIALS AND METHODS59
3.2.1 Experimental site .59
3.2.2 Weather Conditions .59
3.2.3 Germplasm Assembly 0000059
3.2.4 Land Preparation .60
3.2.5 Experimental Design ...60
"
"
"
CHAPTER FOUR00107
MOLECULAR CHARACTERISATION OF 29 ACCESSIONS OF OKRA
(Abelmoschus spp L. Moench) USING INTER-SIMPLE SEQUENCE
REPEATS (ISSR) MARKER00107
4.1 INTRODUCTION000107
4.1.1 Objective of the study112
4.2 MATERIALS AND METHODS113
4.2.1 Experimental Site ..113
4.2.2 DNA Isolation ...113
4.2.3 Preparation of Cocktail for PCR. ..114
4.2.4 Amplification 114
4.2.5 Preparation of Polyacrylamide Gel (PAGE) . 115
4.2.6 Electrophoresis ..115
4.2.7 Data Collection and Analysis 116
4.3 RESULTS00117
4.3.1 PCR Amplification Products .117
4.3.2 Cluster Analysis based on six ISSR primers . 118
4.4 DISCUSSION120
4.4.1 Cluster Analysis 120
4.4.2 Genetic Diversity/polymorphism among Accessions as Revealed by
ISSR Primers . 121
4.5 CONCLUSIONS0122
4.6 RECOMMENDATIONS0123
REFERENCES000125
"
"
CHAPTER FIVE135
TOTAL
FLAVONOID,
PHENOLIC
CONTENTS
AND
ANTIOXIDANT000135
5.1 INTRODUCTION000135
5.1.1 ESTIMATION OF ANTIOXIDANT SCAVENGING ACTIVITY
139
5.1.2 Objective of study .140
5.2 MATERIALS AND METHODS00140
5.2.1 Experimental site...140
5.2.2 Sample preparation141
5.2.3 DPPH Radical Scavenging Assay . 142
5.2.4 Total phenolic content ...143
5.2.5 Total flavonoid content .144
5.2.6 Statistical analyses of data.144
5.3 RESULTS00145
5.3.1 DPPH radical scavenging assay 145
5.3.2 Total flavonoid and phenolic contents of okra accessions 146
5.4 DISCUSSION149
5.4.1 Total flavonoid and phenolic contents of ethanoic and aqueous
extracts of Accessions of Abelmoschus spp ........................................... 149
5.4.2 Total antioxidants activity of 25 accessions of Abelmoschus spp 151
5.5 CONCLUSION00152
5.6 RECOMMENDATIONS153
REFERENCES.154
"
"
CHAPTER SIX000164
DETERMINATION OF ESSENTIAL MINERAL ELEMENTS AND
MUCILAGE IN SOME ACCESSIONS OF OKRA (Abelmoschus spp)164
6.1 INTRODUCTION000164
6.1.1 Okra Production in the world 164
6.1.2 Patronage and Consumption of okra . 165
6.1.3 Proximate composition of okra . 166
6.1.4 Essential mineral elements in okra167
6.1.5 Significance of mineral elements to human ..168
6.1.6 Mucilage172
6.1.7 The Instrumental Neutron Activation Analysis (INAA) Method..174
6.1.8 Objective of the study175
6.2 MATERIALS AND METHODS175
6.2.1 Experimental site...175
6.2.2 Technique for Analysis of Mineral Elements176
6.2.3 Sample preparation for elemental analysis176
6.2.4 Sample irradiation and counting177
6.2.5 Sample preparation and extraction of mucilage 177
6.2.6 Determination of mucilage contents . 178
6.2.7 Data analyses 179
6.3 RESULTS00179
6.3.1 Concentrations of Nine Essential Elements in Accessions of
Abelmoshus spp L. . 179
6.3.2: Association of Essential Mineral Elements. 183
6.3.3 Maximum viscosity of mucilage of Abelmoschus spp L. . 184
"
"
6.4 DISCUSSION0186
6.4.1 ASSESSMENT OF ESSENTIAL MINERAL ELEMENTS IN
OKRA ACCESSIONS ...186
6.4.2 Intensity of Association between Mineral Elements . 188
6.4.3 Mucilage content of Abelmoschus spp L. . 190
6.5 CONCLUSIONS192
REFERENCES..194
CHAPTER SEVEN207
GENERAL CONCLUSIONS AND RECOMMENDATIONS000207
7.1 CONCLUSIONS207
7.2 RECOMMENDATIONS.209
8.0 APPENDICES..00211
"
"
LIST OF TABLES"
"
"
LIST OF FIGURES
Figure 2.1: Stages in the growth and development of okra plant (Valeriana,
4233+015
Plate 1: Photographs of dried fruits of okra accessions..63
Plate 2: Photographs of accessions with fruits0..66
Figure 3.3: A Dendrogram Showing Genetic Relationships among 29
Accessions based on Quantitative and Qualitative Traits using Coefficient of
Euclidean, complete Linked Similarity Matrix.079
Figure 4.1(a): Banding pattern of Genetic diversity studies on okra accessions
numbered 1-15 with CA4 primer.0117
Figure 4.1(b): Banding pattern of Genetic diversity studies on okra accessions
numbered 16-29 with CA4 primer.000000117
Figure 4.2: A dendrogram showing genetic relationship revealed by six
primers among the accessions based on dice coefficient using single link
similarity matrice method000..............................119
"
"
LIST OF ABBREVIATIONS
"
spp
species
et al
and others
NARP
IBPGR
IPGRI
CIP
BSc.
Bachelor of Science
MSc.
Master of Science
MPhil.
Master of Philosophy
AVRDC
PGRRI
CSIR
VEPEAG
DNA
Deoxyribonucleic Acid
RNA
Ribonucleic Acid
ISSR
AFLP
EST
SRAP
QTL
SSR
MAS
RAPD
RFLP
SNP
"
"
PCR
CTAB
TEMED
N, N-bismethylene Acrylamide
APS
PAGE
AMOVA
ITS
ECHO
WAT
FAO
UNESCO
USDA
NIHORT
subsp
sub species
IPM
FAOSTAT
PCA
SLCA
YVMV
NAA
IAA
Indole-3-Acetic Acid
IBA
Indole-3-Butyric Acid
TDZ
Thidizuron
BAP
6-aminobenzylpurine
CP
Coat Protein
NAC
"
"
BNARI
RAMSRI
GHARR-I
ARBC
RTC
GAEC
HPGe
High-Purified Germanium
INAA
MCA
Multi-Channel Analyser
KW
kilowatt
KeV
ANOVA
Analysis of Variance
RCBD
CRD
DMRT
Fwpecpu"ownvkrng"tcpig"vguvu
LoS
Level of significance
Lsd
GS
CIRAD
MT
Metric tonnes
UV
Ultra violet
ROS
SOD
superoxide dismutase
ORAC
FRAP
DPPH
Diphenylpicryl-Hydrazyl
LDL
Low-Density Lipoprotein
"
"
TRAP
TAA
TPC
TFC
dH20
Distilled water
FCR
Folin-Cicalteu reagents
GAE
QE
Quercetin equivalent
AOA
Antioxidant Activity
R2
Coefficient of regression
SD
Standard deviation
HPLC
TLC
LCMS
USB
Metre (s)
cm
Centimetre (s)
mg/g
ug/g
bu
Brabender units
rpm
RDI
"
"
ABSTRACT
A series of investigations were carried out to determine the genetic variability
within 29 accessions of okra (Abelmoschus spp L. Moench) through
characterisation using morphological, biochemical, nutritional and molecular
markers. The goal was to obtain information on key traits of okra germplasm
relevant to breeders and other researchers towards improvement of the crop.
Twenty six (26) indigenous (landraces) and three (3) exotic accessions of
okra were collected from eight regions of Ghana and their morpho-agronomic
traits were evaluated under field conditions at the Biotechnology and Nuclear
Agriculture
Research
fields
using the
International Plant Genetic Research Institute (IPGRI) descriptor list for okra.
The 29 accessions exhibited significant variation in all but two quantitative
traits studied. Block coefficients of variation were extremely low, implying
that results obtained are reliable and repeatable over replications. The 29
accessions were grouped into two major clusters and subsequently into five
sub-clusters based on both quantitative and qualitative characters studied. The
association between pairs of quantitative yield traits in the okra landraces
revealed that flowering and fruiting parameters had significant (P = 0.01)
positive associations. Factor scores of 12 characters contributed substantially
to total genetic variation among the 29 okra accessions studied. The pattern of
clustering did not show distinct association between morpho-agronomic
characters and geographic origin of the collections. The output of the
Principal Components Analysis (PCA) revealed that different characters
contributed differently to total genetic variation. The means of maximum
viscosity values for mucilage extracted from the fruits ranged from 53.0 366.8bu, with three accessions; DKA (366.8bu), Yeji-Local (329bu) and
Amanfrom (316.8bu) recording very high values whilst Cape (53.0bu) had
the least maximum viscosity value. There was low level of polymorphism
detected among all accessions using inter-simple sequence repeat (ISSR)
markers. The accessions, Atomic and Akrave were detected to have
"
originated from a common ancestry. While there was high variability among
Okra accessions for the amounts of flavonoids, phenolics and total
antioxidant activity in the fresh fruits and the quantities were generally high
making okra a good source of natural antioxidants.
Ethanol extraction
Debo
and
Kortebortor-ASR
recorded
the
highest
"
CHAPTER ONE
INTRODUCTION
1.1 GENERAL INTRODUCTION
Okra (Abelmoschus spp L.) also known as okro, is a tropical vegetable crop,
grown throughout Africa (Schippers, 2000; Norman, 1992; Sinnadurai, 1992;
Kochhar, 1986), Asia and the Americas (Siemonsma and Kouame, 2004;
Tindall, 1983). It is a semi-fibrous annual herb (Bennett-Lartey and OtengYeboah, 2008; Oyelade et al., 2003; Rice et al., 1990), and belongs to the
family Malvaceae (Rice et al., 1990; Tindall, 1983). The plant is robust and
erect, ranging between 1 to 2m in height, with simple leaves which are
alternate and palmately veined. The flowers are regular and solitary, with
superior ovaries and numerous stamens. It is widely cultivated for its
immature fruits and seeds.
The name "okra", most often used in the United States and the Philippines, is
of West African origin. Okra is often known as Lady's Fingers outside the
United States. In Bantu language, okra is called kingombo, quiabo in
Portuguese, quimbomb or guigamb in Spanish, gumbo in French, bhinde or
bhendi in India. It is called Molondrn in the Dominican Republic, and
"
India, Costa Rica, Nigeria and Ghana are among the major producers of the
crop (FAOSTAT, 2012; NARP, 1993). The Brong Ahafo, Northern, Volta,
Greater Accra and Central regions are the bulk producers in Ghana (NARP,
1993).
The immature green pods and fresh leaves are used locally as
potherbs. Locally, they are mainly employed as a boiled vegetable, they can
be stir-fried, battered and deep-fat fried, microwaved, steamed, baked,
grilled, blanched and processed as a frozen (plain or breaded), pickled, or
canned product (Oppong-Sekyere et al., 2012). Dried fruits of okra, as slices
or in powder form are often stored and used in stews and soups. The fresh
okra fruits are high in vitamins A and C; and in calcium (NARP, 1993).
"
"
Intra as well as interspecific variations do exist in okra in different phytogeographic areas of Ghana (Anonymous, 2010; Ahiakpa, 2009). However,
like most tropical crops, very little research attention has been devoted to this
crop, particularly in Ghana (Ahiakpa, 2009). Duplications are masked by the
use of local names. Previous works are limited to the use of morphological
traits in cultivar identification and characterisation (Oppong-Sekyere, 2011;
Essilfie et al., 2010; Bennett-Lartey and Oteng-Yeboah, 2008). Hence, there
is the need to also carry out biochemical, nutritional and molecular
characterisation to fully describe the available germplasm of okra (Ahiakpa,
2009; Lester et al., 1990). This information will consequently enhance rapid
improvement of the crop.
"1.1.2
"
In addition, plant genetic resources are conserved for future use. Using them
is only possible if their characteristics or attributes are known in detail and
their utilisation visualised (Jaramillo and Baena, 2000). Such knowledge and
visualisation can be achieved only through the study of the morphological,
structural, and functional attributes of germplasm, as the carriers of all the
hereditary characteristics of any species.
These methodologies (molecular and biochemical) help locate genes of
interest with greater accuracy but do not evaluate the effect of the
environment on the expression of those genes (Westman and Kresovich,
1997). Accordingly, they do not replace but complement morpho-agronomic
characterisation and evaluation (IPGRI, 2004; IPGRI and CIP, 2003a&b).
"
"
The local landraces in Ghana have long maturity periods and short
harvesting duration. They are of poor nutritional quality, non-standard in
shape, colour and size, making them unfit for the Ghanaian okra vegetable
export market (Oppong-Sekyere, 2011). This has a consequential impact of
causing a tremendous reduction in the per capita income of the nation. It is
therefore important that plant breeders develop improved varieties of okra,
for adoption by Ghanaian vegetable farmers and for the export market.
Varieties that are perennial in growth habit and at the same time combine
higher yields and early maturity with longer harvest duration and also
resistant to diseases and pests would be ideal for the okra vegetable industry
in Ghana (Oppong-Sekyere et al., 2012).
Such varieties must also possess improved fruit size, shape and colour,
quality characteristics, very much desired in the okra export market (Boateng,
2011.per comm.). Currently, this vegetable is a non-traditional export crop in
Ghana but interestingly, only exotic varieties meet export specifications."
Vjgtghqtg." vjgtg" ku" vjg" need to develop local varieties to expand production
base. The first step towards attaining this objective involves collection and
thorough characterisation of available germplasm of the vegetable.
improvement programme of the crop (Ahiakpa, 2009; Lester et al., 1990; and
Charrier, 1983).
"
"
REFERENCES
References for CHAPTER ONE are merged with those for CHAPTER
TWO.
"
CHAPTER TWO
LITERATURE REVIEW
2.1 ORIGIN, DISTRIBUTION AND UTILISATION
Okra (Abelmoschus spp L.) is a member of the family Malvaceae same as
cotton. It is of African origin (ECHO, 2003; Purseglove, 1987), discovered
around Ethiopia during the 12th century B.C and was cultivated by the
ancient Egyptians (Purseglove, 1987). Currently, the crop is grown as a
popular vegetable throughout the tropical and sub-tropical regions of the
world (Kumar et al., 2010).
Hamon and van Sloten (1989) classified the cultivation areas of okra into four
climatic zones. These include; desert (village or oasis cultivation), sahelian
(north of latitude 12 oN), savannah (between latitudes 8 oN and 12 oN) and
rain forest climatic zones. Abelmoschus esculentus is primarily distributed
throughout the intermediate savannah zone between the rain forest and the
arid sahel. The species is less frequently found in the rain forest zone but is,
on the other hand fairly well represented in the sahel zone. On the contrary,
the West African Taxon (WAT), A. caillei does not occur in the sahelian zone
since it has a long life cycle and usually requires abundant, continuous
rainfall. Hamon and van Sloten, (1989) stated that since a natural interspecific
hybrid of the two cultivated species occurs in the central part of Sudan, A.
caillei is possibly more widely distributed than currently known.
"
the leaves of okra can eliminate oxygen free radicals, alleviate renal tubularinterstitial diseases, improve renal function and reduce proteinuria
(Siemonsma and Kouame, 2004).
In West Africa, the plant is cultivated as a vegetable crop and the leaves, buds
and flowers are often eaten (Siemonsma and Kouame, 2004; Tindall, 1983).
The fruits may be dried, stored and powdered for use in soups in the dry
season when fresh fruits are scarce (Oppong-Sekyere, 2011; Siemonsma and
Kouame, 2004; Oyolu, 1977). The immature fruits are used as boiled or fried
vegetable (Tindall, 1983). The roots can be used to cure syphilis (FAO,
1988). The mucilage in pods (a glutinous substance) is used in thickening
soups and stews (National Academies Press, 2006; Woolfe et al., 1977). The
matured seeds contain about 20% of edible oil (Tindall, 1983) and can be
used for biodiesel production (Anwar et al., 2010).
"
"
"2.2
"
green
to
dark
brown
upon
maturity
"
"
were originally collected from six different countries in the region. Six
duplicate accessions were discovered while accession TOT7444 distinguished
itself from the other two okra species, an indication that it might belong to a
different species. This recent study at molecular level emphasises the need for
a deeper study into the variable polymorphism at chromosomal level in the
genus Abelmoschus.
Table 2.1:
Potential
of
recombination breeding
involving
two
Abelmoschus spp"L""
Species
A. esculentus (common okra)
95% cultivated area
Cytogenetics
Amphidiploid (2n=130-140): A.
tuberculatus or A. ficulneus (2n-5860) x unknown?
Contrasting Traits
Poor adaptation in humid zone,
more susceptible to biotic
stresses, less vigorous, short life
cycle (suitable for short rainy
season areas), usually day neutral,
cultivated in both rainy (rain fed)
and dry (irrigated) season
"
Diversity in pod shape, size and flowering behaviour account for most of the
variation between the genotypes of West and Central African origin
(Duzyaman, 1997) and scope for further gain in pod yield per plant is limited
because of low phenotypic and genotypic variability (Ariyo, 1990a). To break
the yield barrier in existing genotypes of common okra (A. esculentus) and
breed for different market types, a hybridisation-based breeding strategy
would be desirable.
Short day length stimulates flowering of most cultivars (Martin et al., 1979).
Flowering begins at a very early stage of growth at day lengths of less than 11
hrs; under long days, the flower buds tend to abort (Chauhan, 1972). Okra
requires adequate soil moisture throughout its growing period for optimum
growth and yield (Norman, 1992).
"
(a)Seeding stage
Figure 2.1: Stages in the growth and development of okra plant (Valeriana, 2011)
(a) Seeds are numerous, gray to black in colour and about 3-6 mm in diameter. Seeds germinate about 5-7 days
after sowing.
(b) This is between one to two weeks from seed germination. Seedlings have at least 3-4 leaves with a height
approximately 12-18cm.
"
(c) This occurs three to four weeks from seed germination. Leaves are bigger and the plant has more than eight
leaves. Length of stem between leaves is longer. The leaves are spreading and spirally arranged.
(d) Plants start to flower at five weeks from seed germination. Yellow solitary flowers are in the leaf axils. Okra
usually flowers within 40-90 days after sowing. Flower opens in the morning. Fruits or pods are green,
cylindrical to pyramidal capsule 5-35cm long and 1-5cm in diameter.
It takes about one month from anthesis to fruit maturity. Mature green pods
turn brown and dry. On the seed crop, vegetative growth stops soon after
anthesis, all assimilates are partitioned to the reproductive parts of the plant
(Kumar et al., 2010). Flower initiation and flowering are delayed at higher
temperatures, indicating a positive correlation between temperature and
number of vegetative internodes (Norman, 1992).
"
"2.6
Okra has perfect flowers, thus male and female reproductive parts are in the
same flower and are self-pollinating. If okra flowers are bagged to exclude
"
"
pollinators, 100% of the flowers will set seed. It has been demonstrated
experimentally that there is no significant difference in fruit set under openpollinated, self-pollinated (by bagging alone) and self-pollinated (hand
pollination of bagged flowers) plants, indicating that it is primarily a selfpollinated crop (Valeriana, 2011; Datta and Naug, 1968).
2.7 RATOONING
Some farmers practise ratooning when seeds are scarce and when the buying
period is extended. Instead of establishing a new crop, farmers cut the main
stem of the first crop. They then apply herbicides to kill the weeds before
applying fertiliser and irrigating (Schippers, 2000). Some farmers, however,
apply sodium nitrate to enhance flowering. It takes less than a month to
harvest from the ratoon crop (Valeriana, 2011). However, pods from ratoon
crop are of less quality and fewer than the former crop (Camciuc et al., 1996).
This is so because there would be more branches per plant to provide the
nutrients.
"
Incidence of more larvae of fruit worm and cutworm (Agrotis spp.) has been
monitored from ratoon crops. The larvae of these pests come from the first
crop and they pupate in the soil (Anonymous, 2010). Ratoon crops do not
require tillage and therefore soil is not disturbed. The duration of time from
pupa to adult, egg laying and hatching of eggs into larvae coincides with that
time the ratoon crop is producing more leaves and flower (Siemonsma and
Kouame, 2004). Besides the larvae, other insect pests will transfer to the
ratoon crops when they have no more food from the first crop in the
neighbouring fields (Midmore et al., 2005).
"
al., 1978), calcium and iron (Savello et al., 1980), which are integral to health
in humans (Kumar et al., 2010).
2008),
mucilagenous
effect
(Ameena
et
al.,
2010),
"
1,769 from West Africa (Oppong-Sekyere, 2011; Hammon and van Slotten,
1989). Okra, therefore, is far more prominent in West Africa than in other
parts of the world (Omonhinmin and Osawaru, 2005). The genetic diversity
of okra is clearly shown by the wide range of morphological characteristics
displayed by the taxon in different ecogeographical, edaphic and
environmental conditions (Omonhinmin and Osawaru, 2005).
"
There exists a wide genetic diversity among cultivated species of the genus
Abelmoschus (Omonhinmin and Osawaru, 2005; Bisht et al., 1995) and are
highest amongst species found in countries such as Turkey and India (Bisht et
al., 1995). Porter et al., (1974) reported that large morphological variability
abounds in the tropics suggesting adequate research into the germplasm
structure for development of hybrids with specific ecological adaptation.
Variability is more prominent in days to flowering, plant height and various
fruit characteristics among okra germplasm. Hence, these traits could be
important in differentiating varieties of A. esculentus (Ariyo and Odulaja,
1991). Within species variation among 30 West African genotypes were
found to be considerably large based on phenotypic assessment (Ariyo,
1993). Gulsen et al. (2007) however stated that, the West African sub region
being the second largest producer of okra may have considerable level of
genetic diversity as in many other important crop species, which calls for
thorough research investment to fully exploit these potentials in the crop.
"
Genetic diversity has been reported in the West African and South Asian
accessions of okra (Bisht et al., 1995; Ariyo, 1993). A good understanding of
genetic variability in the different characters of okra would be a useful tool in
the genetic improvement of the crop. This will enhance the identification of
useful genes and their behaviour as an aid in hybridisation programmes. The
characterisation of a range of genetic variability among genotypes is pivotal
to the maintenance and further acquisition of germplasm resources even as
accessions from diverse origins are needed as parent stocks for the
development of improved varieties (Aremu et al., 2007).
"
"2.10.1
For the improvement of the yield and other desirable characters, a knowledge
of the magnitude of variation in the available genotypes, the relation of
characters with yield, extent of environmental influences on these and the
heritability of the materials are essential (Saifullah and Rabbani, 2009).
Morphological characterisation is the most dominant in okra germplasm.
These include recent assessment of genetic diversity in a collection of 22
Ghanaian okra germplasm (Abelmoschus spp L.) using morphological
markers (Oppong-Sekyere, 2011); morphological classification of genetic
"
"
morphological
West
Kresovich et al. (1992) showed that molecular markers are of great value in
genetic resource management as quick, cost-effective and reliable methods
for identification, measurement of variation and determination of similarity at
the intra-specific level. Reports on marker development in okra are very
scanty and have been mostly limited to characterisation of cultivars.
"
"
Marker aided selection (MAS) for various traits in okra germplasm in Turkey
has been suggested using sequence-related amplified polymorphism (SRAP)
(Gulsen et al., 2007). Recently, twenty okra accessions from Burkina Faso
were analysed using 16 primers designed to amplify SSR regions of
Medicago truncatula. Two accessions were found distinct from the other 18,
based on the presence of an unique 440 bp fragment generated primer MT-27
and also based on presence of hairs on fruits and delayed maturity of these
two accessions (Sawadogo et al., 2009).
"
"
Okra has huge potential for enhancing livelihoods in urban and rural areas
and to several stakeholders (Kumar et al., 2010; National Academies Press,
2006). It offers a possible route to prosperity for small-scale and large-scale
producers alike and all those involved in the okra value chain, including
women producers and traders. Both pod skin (mesocarp) and seeds are
excellent sources of zinc (80 mg/g) (Cook et al., 2000; Glew, 1997). Okra
seed is mainly composed of oligomeric catechins (2.5mg/g of seeds) and
flavonol derivatives (3.4 mg/g of seeds), while the mesocarp is mainly
composed of hydroxycinnamic and quercetin derivatives (0.2 and 0.3mg/g of
skins). Pods and seeds are rich in phenolic compounds with important
biological properties like quartering derivatives, catechin oligomers and
hydroxycinnamic derivatives (Arapitsas, 2008). These properties, along with
the high content of carbohydrates, proteins, glycol-protein, and other dietary
elements enhance the importance of this foodstuff in human diet (Arapitsas,
2008; Manach et al., 2005).
In addition, fresh okra pods are the most important vegetable source for
viscous fibre, an important dietary component to lower cholesterol (Kendall
"
"
and Jenkins, 2004). Okra seed oil has potential hypocholesterolemic effect
(Rao, 1985; Ramu, 1976). The potential for wide cultivation of okra for
edible oil as well as for cake is very high (Rao, 1985). Okra seed flour could
also be used to fortify cereal flour (Adelakun et al., 2009). Okra mucilage has
potential for use as food, non-food products, and medicine (Ndjouenkeu et
al., 1996; BeMiller et al., 1993). Okra mucilage is used in Asian medicine as
a protective food additive against irritating and inflammatory gastric diseases.
Researchers worldwide are looking for already characterised available
germplasm of the plant to explore all these enumerated potentials and exploit
them fully to the benefit of society (Lengsfeld et al., 2004).
"
Okra offers many production possibilities; however, there are limited studies
conducted on its germplasm due to limited resources devoted to the species
by national and international research institutes. Additionally, recent research
goals are geared towards fast maturing types well suited to tropical heat, and
humidity conditions and require adequate knowledge of available germplasm
of the plant to facilitate full exploitation of these potentials (Kumar et al.,
2010). Most crop geneticists agree that enrichment of the cultivated gene pool
will be necessary to meet the challenges that lie ahead (Tester and Langridge,
2010).
"
Qmtcu" rqvgpvkcn" hqt" kpfwuvtkcn" wug" cpf" eqpvtkdwvkqp" vq" gpjcpeg" nkxgnkjqqfu"
cannot be underestimated. Okra offers many production possibilities;
however, there are limited studies conducted on okra biology and production
due to limited resources devoted to the species by national and international
research programmes. With huge prospects for emerging agro-industry
worldwide, demand of the plant for specific use such as oil, mucilage
material, paper making material, bioabsorbent, pharmaceuticals and its other
untapped potentials for industrial use, makes it imperative for breeding
"
"
Fast maturing types are well suited to tropical heat, humidity and also to dry
(rain-fed) and hot (Sudano-Sahelian) conditions. Pods contain high amounts
of usable dietary fibre and they are often dried, stored and consumed as
soup/sauce much like a staple food. Half a cup of the cooked pods (fresh)
provides about 10% of the recommended levels of vitamin B6, folic acid and
vitamins A and C (Camciuc et al., 1996) and is sought after in food
processing industries.
"
The seed (usually consumed with pods) protein is distinct from both cereals
and legumes. Because it can easily be dried and stored for long periods
(unlike perishable vegetables), producers and processors are better able to add
value and take advantage of seasonal fluctuations in price. Besides pod yield,
the foliage and stems can weigh up to 27 tons per hectare of biomass, which
is likely to become useful with fuel prices increasing globally and new
technologies promising efficient conversion to liquid fuels. It is worth noting
that okra stems generate considerable heat without sparks, excessive smoke,
or bad odours. The potential for non-vegetable use include paper pulp, like its
close relative kenaf, oil seed, sacks and ropes, plasma replacer and
suspending agent in medicine (National Academies Press, 2006).
"
"
"
and photoperiod sensitivity and insensitivity are among the most devastating
factors that militate against okra production in the West African sub-region.
Lack of improved genotypes has however been identified as the most
pressing challenge that requires prioritised attention as a first step towards
improving the crop in the sub-region (AVRDC, 2010).
There are few varieties across the sub-region, well-adapted to the respective
geographical climates but are generally low yielding, have long maturity
periods yet short harvesting periods and often with narrow genetic base
(Schippers, 2000). Most of these are also highly susceptible to pest and
disease attacks, highly sensitive to photoperiodic fluctuations, and have nonuniform pod shape and size, varied nutritional constituents, undesirable
features (spines) for handling and unwholesome for export. This problem can
be solved through thorough and comprehensive germplasm screening,
evaluation, purification and promotion of better and superior adapted lines
that meet market types (specific market demands such as mucilage, balanced
nutritional composition) and preferences, high yielding with shorter maturity
and longer harvesting durations and generally adaptable to the sub-regional
agro-climatic and ecological conditions.
"
Stem borers (Earias spp) damage shoots at early vegetative stage. The plants
then develop branches to compensate for the damage by stem borers. Unlike
"
"
Damping off at seedling stage can cause tremendous losses unless most of the
seeds sown are treated with fungicides. Leafhoppers, aphids and whiteflies
attack at seedling to early vegetative stage and transmit the yellow vein
mosaic virus. Infected plants produce poor quality pods. Rapid increase in
leafhopper population during the dry season causes hopperburn in okra
especially when there is no source of yellow vein mosaic virus in
neighbouring fields. Leaves turn red and eventually dry up due to feeding by
high density of sucking insects at the vegetative stage predisposing them to
leaf curl and defoliation (De Lannoy, 2001).
Cercospora leaf spot and powdery mildew (Oidium spp) are two fungal
diseases from the late vegetative to the reproductive stage. The two are fungal
infections and spread rapidly on the field due to crowded and overlapping
broad leaves of plants. Besides wind, spreading fungal spores to plants,
people harvesting daily and passing along okra rows are also responsible for
the widespread infection in the field. Farmers remove and drop the old yellow
leaves with Cercospora leaf spot to reduce infection but usually these leaves
are not properly disposed off and the sources remain on the field (Norman,
1992).
"
"
"
"
Shoot and fruit borers (Earias spp) are the most destructive pests in okra
crop. Efforts have been made to develop insect resistant okra varieties by
incorporating cry1Ac gene in okra from a bacterium mainly Bacillus
"
"
NAA and 0.5 mg L-1 TDZ. Highest percentage shoots regeneration and
means number of callus regeneration per mass callus was obtained at 2.0 mg
L-1 BAP and 0.1 mg L-1 IBA, root regeneration was observed at 1.5 mg L-1
NAA. Eighty percent (80%) of the regenerated plantlets survived and showed
new leaves development under ex vitro condition (Anisuzzaman et al., 2008).
This protocol would be useful for creation of somaclonal variation and
enhancing utilisation of transgenic approaches for varietal improvement of
okra.
"
REFERENCES
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Venter, M. V. (2009). Chemical composition and the antioxidative properties
of Nigerian Okra Seed (Abelmoschus esculentus Moench) Flour. Journal of
Food Technology, 47 (6): 1123-1126.
Aladele, S.E., Ariyo, O.J. and Robert de, L. (2008). Genetic Relationship
among West African Okra (Abelmoschus caillei) and Asian genotypes
"
"
Ameena, K., Dilip, C., Saraswathi, R., Krishnan, P. N., Sankar, C., Simi, S. P.
(2010). Isolation of the mucilages from Hibiscus rosasinensis (linn.) and
Okra (Abelmoschus esculentus linn.) and studies of the binding effects of the
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Anisuzzaman, M., Jarin, S., Naher, K., Akhtar, M.M., Alam, M.J.,
Khalekuzzaman, M., Alam, I. and Alam, M.F. (2008). Callus Induced
Organogenesis in Okra (Abelmoschus esculentus L. Moench). Asian Journal
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Anwar, F., Umer, R., Ashraf, M., Nadeem, M. (2010). Okra (Hibiscus
esculentus) seed oil for biodiesel production. Journal of Applied Energy, 87
(2010): 779785.
Arapitsas, P. (2008). Identification and quantification of polyphenolic
compounds from okra seeds and skins. Journal of Food Chemistry, 110 (4):
1041-1045.
Aremu, C.O., Adebayo, M.A., Ariyo, O.J., Adewale, B.B. (2007).
Classification of Genetic diversity and choice of parents for hybridization in
cowpea Vigna unguiculata (L.) Walp for humid savanna ecology. Afr. J.
Biotechnol. 6(20): 2333-2339.
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Vegetable
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Problems
with
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Okra.
th
July
2011).
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Golan- Goldhirsh, A. (2003). Genetic variability in Turkmen populations of
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"
"
BeMiller J.N., Whistler R.L., Barkalow D.G., Chen C.C. (1993). Aloea, chia,
flax seed, okra, psyllium seed, quince seed, and tamarin gums. In: Industrial
Gums, Whistler RL, BeMiller JN (eds.), Academic Press, New York, pp.
227-256.
Bennet-Lartey, S.O. and Oteng-Yeboah, A. A. (2008). Ghana Country Report
on the State of Plant Genetic Resources for Food and Agriculture. CSIR,
Plant Genetic Resources Research Institute, (PGRRI), Bunso, Ghana. pp.1533.
Bisht I.S., Mahajan R.K., Rana R.S. (1995). Genetic diversity in South Asia
okra. (A. esculentus) germplasm collection. Ann. Appl. Biol. 126: 539-550.
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"
rd
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Cook, J.A., Vander Jagt. D.J., Pastuszyn, A., Mounkaila, G., Glew, R. S.,
Millson M., Glew, R.H. (2000). Nutrient and chemical composition of 13
wild plant foods of Niger. J. Food Comp. Anal, 13: 83-92.
Duzyaman, E. (1997). Okra: Botany and Horticulture. Hortic. Rev. 21: 4172.
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"
"
Glew, R.H., Vander Jagt, D.J., Lockett, C., Grivetti, L.E., Smith, G.C.,
Pastuszyn, A., Millson, M. (1997). Amino acid, fatty acid, and mineral
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10:205-217.
Gulsen, O., Karagul, S., Abak, K. (2007). Diversity and relationships among
Turkish okra germplasm by SRAP and phenotypic marker polymorphism.
Biologia Bratislava, 62: 41- 45.
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"
IITA. (2001). West Africa plant genetic resources update. The International
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MARA
(UiTM)
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International
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B.A. (1992). Characterisation of genetic identities and relationships of
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Lamont, W.J. (1999). Okra-A versatile vegetable crop. Hort Technol. 9: 179184.
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Tomoda, M., Shimizu, N., Gonda, R., Kanari, M., Yamada, H., Hiki, H.
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Yildiz, M. U., Sedat, C., Musa, O., Haydar, H. (2005). A study on some
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valsartan
in
treatment
of
early
diabetic
nephropathy
with
"
"
CHAPTER THREE
3.1 INTRODUCTION"
The primary objectives of okra germplasm characterisation have generally
been to identify high yielding genotypes with resistance to yellow vein mosaic
virus (YVMV), fruit borer (Spodoptera spp.), jassid (Cicadellidae) and higher
vitamin C content in the species that can be utilised for the improvement of
the crop (Nwangburuka et al., 2011; Bisht and Bhat, 2006). Morphological
markers are the traditionally accepted and proven tools as a first step, among a
host of other techniques, for characterisation of plant germplasm. They
constitute the most readily available technique. Thus, published descriptor
lists are readily available for most major crop species including okra. The
technique can be carried out in situ (on-farm), it is relatively inexpensive, easy
to carry out with little skills and minimum facility requirements for use
(Hoogendijk and Williams, 2001).
Ochrosol
(Ferric
Acrisol)
derived
from quartzite
Schist
(FAO/UNESCO, 1994).
"
Accession
Ashanti region
Central region
Cape
Eastern region
Brong-Ahafo region
Western region
Juaboso
Volta region
Akrave, Kpeve
drawing lots to avoid bias and each plot was well labelled.
"
No fertiliser was applied, but weeds were controlled and other agronomic and
management practices were carried out. Weeding was done fortnightly. The
rainfall pattern during this period was quite consistent and regular hence there
was no need for supplementary water to support plant growth and
development.
"
which include the following parameters and were grouped into four growth
stages of the plant;
(a) Vegetative characters: general aspect of the plant, branching type (BRT),
stem pubescence, stem colour, leaf shape, leaf colour and total number of
leaves per plant (NLPP). Data was taken on these characters prior to first
fruiting of all accessions ensuring that all accessions within each block
receive uniform treatment. Five plants were randomly selected for quantitative
traits such as number of leaves and this was done after each flowering. Data
on qualitative traits were taken based on confirmation from each block.
(b) Inflorescence characters: number of epicalyx segments (NES), shape of
epicalyx segments (SES), persistence of epicalyx segments (PES), petal
colour, and colouration of petal base. These characters were recorded during
the flowering stage of each accession in all blocks. This was done on five
plants out of the eight plants.
(c) Reproductive characters: days to 50% germination (FPGer), maximum
plant height (cm) (MPH), stem diameter at the base (mm) (STB), maximum
number of internodes (MNI), days to 50% flowering (FPFl), first flowering
node (FFN), first fruit-producing node (FFPN), total production of pods, and
number of pods per plant (NPPP). A tape measure was employed in taking
plant height at the fruiting stage for five data plants. Stem diameter at the base
was taken at maturity level of the accessions using a pair Vernier Callipers.
(d) Fruit characters: position of fruit on main stem (PFMS), fruit colour, fruit
length at maturity, length of peduncle, fruit shape, number ridges per fruit
(NRPP), fruit pubescence and number of days to 50% fruiting (FPFr).
Position of fruit on main stem was determined on five data plants prior to
harvesting of fruits. Measuring rule was used for measuring length of pods
"
and peduncle of pods after harvest. Number of ridges was done by counting
the quantity of ridgeline or natural striations through the fruit and then coded
accordingly. This was done on five pods of each accession. Data on fruit
pubescence was taken by a visual assessment of hairiness or smoothness of
rqfu"cpf"c"rtcevkecn"hggn"qp"jctxguvgf"htguj"htwkvu0
(e) Seed characters: shape of seed, aspect of the surface, average number of
seeds per pod (ASPD), weight of 1000 seeds (g), (TSwt). All seed characters
were measured after harvest of the accessions. Harvested pods were further
dried and seeds extracted and then dried separately to reduce moisture content
to an appreciable level. Seeds per pod were counted for each accession; one
thousand (1000) seeds per accession were weighed using a digital scale.
"
3.3 RESULTS
3.3.1 Morphological Traits of dry pods of Accessions
The photographs below show phenotypic characteristics of fruits of
the various accessions at maturity.
DKA
Labadi
Yeji-Local
Asontem nv.
Cs-Legon
Mapelega
"
"
Cape
Volta
Juaboso
Debo
Akrave
Kpeve
Asontem-ER
Legon fingers
Figure.2: contf
"
"
Asontem-BAR
Asante type II
Amanfrom
Asontem-ASR
Atomic
Agric type I
Kortebortor-BAR
Asontem-GAR
Plate.2: eqpvd
"
"
"
Plate 1: Variation in Vegetative and Reproductive Characteristics of some Accessions of Okra.
"
"
Rncvg"3"*Eqpvf+<
"
"
In addition, Kortebortor-BAR had the thickest stem diameter, but this was not
significantly different from Amanfrom. Asontem-ASR had the thinnest stem
diameter but was not significantly different from Cape, Debo and Clemson
Spineless.
"
DESIGNATION
MPH
MNI
FFN
NLPP
ASPD
STB
NPPP
FPGer
FFPN
FPFl
FPFr
TSwt
APP
Nkran Nkuruma
170.78a
19.00a
23.00a
20.00de
18.25r
9.98c
23.50ab
53.50ghi
10.75bc
50.00e
6.00j
69.43b
40.50e
128.53
18.00
14.50
jk
9.85
17.50
de
51.25
mn
ab
47.00
8.00
53.92
11.00o
116.33
bc
18.00
16.00
hij
8.15
gh
16.25
de
53.00
ijk
47.00
9.50
50.81
21.50lm
113.40
cd
18.00
16.00
12.50
lm
9.00
14.25
ef
52.00
klm
39.00
8.00
64.41
22.00l
103.95
cde
16.78
34.00
9.55
de
18.25
cd
78.00
39.00
62.14
37.25f
102.53
def
15.02
13.75
kl
5.93
op
18.25
cd
53.75
gh
38.50
59.03
52.50b
18.50
13.75
kl
25.25
92.00
82.00
53.83
57.50a
46.71r
44.50c
Asontem-BAR
Asante type II
Asontem-GAR
Yeji-Local
Cape
Kortebortor-
99.28
ef
13.00
9.00
8.00
l
e
11.75
19.00
21.25
pq
22.75
op
41.25
63.00
43.00
28.5
lmn
11.03
11.75
9.25
8.0
fghij
8.25
9.5
cdefg
fghij
cdefg
9.75
bcdef
10.00
11.75
cd
8.00
BAR
Juaboso
94.75efg
17.50d
18.00c
26.75c
46.25c
8.10hi
14.50ef
53.30hij
8.00fghij
47.00g
10.25ef
Labadi
92.55efgh
18.00c
11.00i
18.50ef
37.00f
8.78f
90.08fghi
13.67j
10.25j
17.00fgh
37.00f
6.38 lm
7.50hi
51.00mn
7.75ghij
40.00n
13.00b
46.71r
28.25h
17.00de
51.00mn
7.75ghij
44.00k
12.00c
52.45l
26.00ij
LoS
**
**
**
**
**
**
**
**
**
**
**
**
**
BE
ns
ns
ns
ns
ns
**
ns
ns
**
ns
ns
ns
ns
TCV (%)
11.9
1.1
1.5
6 .1
3.6
2.6
18.0
2.1
16.7
1.2
1.3
0.5
3.8
BCV (%)
1.3
0.3
0.3
1.7
0.5
1.0
2.9
0.2
6.3
0.42
0.3
0.1
1.0
ns kpfkecvgu"pqp"ukipkhkecpeg"cv"vjg"r2027"ngxgn.","kpfkecvgu"ukipkhkecpeg"cv"vjg"r2027"ngxgn"cpf",,"kpfkecvgu"jkij"ukipkhkecpeg"cv"r2023"ngxgn0"NqU"?"ngxgn"qh"
significance, BE = block efficiency, TCV = treatment co-efficient of variation, BCV = block co-efficient of variation and Mean represent average of the
individual characters measured for all accessions under consideration. NV = Northern Version, MPH = Maximum plant height (cm), MNI = Maximum number
of internodes, FFN = First flowering node, NLPP = Number of leaves per plant, ASPD = Average seeds per pod, STB = Stem diameter at base (mm), NPPP =
Number of pods per plant, FPGer = 50% Germination, FFPN = First fruit producing node, FPFl = 50%Flowering, FPFr = 50%Fruiting, TSwt = 1000 seed
weight (g), APP = Average pods per plant.
"
Tcdng"504"*Eqpvf+<"
DESIGNATION
MPH
MNI
FFN
NLPP
ASPD
STB
NPPP
FPGer
FFPN
FPFl
FPFr
TSwt
Asontem-ASR
89.75fghi
15.87g
9.00l
13.75kl
33.00h
5.25q
15.5de
55.00f
8.00fghij
42.00l
8.00i
41.32w
15.75n
Asontem NV.
83.70ghij
17.00e
17.00d
21.00d
46.00c
7.55k
23.25ab
51.25mn
7.0 0ijk
41.00m
12.00c
63.54e
58.25a
Amanfrom
82.85ghij
17.87c
13.00g
26.00c
51.00b
10.53b
21.50bc
89.00c
13.50a
71.50d
10.50e
56.02i
30.50g
17.00
11.00
15.75
ij
31.25
jk
47.00
12.00
33.92
51.50b
18.93
7.00
18.75
ef
31.75
hi
47.00
11.50
53.24
28.00h
18.00
9.00
27.75
mn
47.00
12.00
58.98
42.25d
17.00
10.00
29.00
lm
32.00
10.00
51.87
21.50lm
16.00
11.00
37.50
45.00
10.00
74.95
25.00jk
16.46
7.00
20.00
45.00
10.00
67.37
12.00o
14.37
7.00
37.00
40.00
10.00
56.33
hi
21.50lm
Volta
Debo
Agric type I
Indiana
Legon fingers
Mamolega
Cs- Legon
79.5
hijk
77.7
ijkll
76.05
71.0
jk
jklmn
72.63
jklm
62.13
nop
64.75
lmno
11.0
mn
12.50
lm
29.00
12.75
20.75
6.55
5.93
op
8.10
hi
7.85
ij
6.35
lmn
6.15
mno
8.40
17.50
de
15.25
de
10.00
ghi
16.25
de
17.75
de
8.25
ghi
17.00
de
53.00
ijk
10.00
51.25
mn
jk
54.25
fg
39.25
51.00
mn
49.00
51.5
lmn
6.50
10.25
bcde
bcd
8.80
defghi
8.80
defghi
8.80
defghi
7.00
ijk
APP
LoS
**
**
**
**
**
**
**
**
**
**
**
**
**
BE
ns
ns
ns
ns
ns
**
ns
ns
**
ns
ns
ns
ns
TCV (%)
11.9
1.1
1.5
6 .1
3.6
2.6
18.0
2.1
16.7
1.2
1.3
0.5
3.8
BCV (%)
1.3
0.3
0.3
1.7
0.5
1.0
2.9
0.2
6.3
0.42
0.3
0.1
1.0
ns kpfkecvgu" pqp" ukipkhkecpeg" cv" vjg" r2027" ngxgn." ," kpfkecvgu" ukipkhkecpeg" cv" vjg" r2027" ngxgn" cpf" ,," kpfkecvgu" jkij" ukipkhkecpeg" cv" r2023" ngxgn0" NqU" ?" ngxgn" qh"
significance, BE = block efficiency, TCV = treatment co-efficient of variation, BCV = block co-efficient of variation and Mean represent average of the individual
characters measured for all accessions under consideration. NV = Northern Version, MPH = Maximum plant height (cm), MNI = Maximum number of internodes, FFN
= First flowering node, NLPP = Number of leaves per plant, ASPD = Average seeds per pod, STB = Stem diameter at base (mm), NPPP = Number of pods per plant,
FPGer = 50% Germination, FFPN = First fruit producing node, FPFl = 50%Flowering, FPFr = 50%Fruiting, TSwt = 1000 seed weight (g), APP = Average pods per plant.
"
Vcdng"504"*Eqpvf+<"
DESIGNATION
MPH
MNI
FFN
NLPP
ASPD
STB
NPPP
FPGer
FFPN
FPFl
FPFr
TSwt
APP
Mapelega
62.40nop
13.16k
10.25j
18.00fg
38.00f
6.08no
6.50i
47.25p
8.00fghij
32.00r
10.00f
56.41h
20.25m
Wune mana
62.13nop
13.00k
6.00o
16.75ghi
12.25s
6.30lmn
6.50i
59.00e
4.25l
49.00f
10.00f
50.23o
7.00p
15.00
43.73
52.50b
12.00
48.12
27.00hi
44.86
15.00n
52.75
22.50l
45.50s
24.25k
DKA
Agric short fruit
Kpeve
Kortebortor-
60.70
nopq
59.00
opqr
56.68
pqrs
48.50
qrs
2.00
17.00
12.32
11.23
17.00
6.00
8.00
7.00
21.50
13.75
kl
17.25
fghi
16.50
ghi
31.50
ijk
23.75
35.00
30.00
kl
8.90
7.80
jk
7.55
9.80
cd
10.25
8.00
ghi
11.25
6.70
gh
fg
hi
125.00
52.50
jkl
55.00
50.75
7.50
hijk
7.00
ijk
9.00
defghi
8.00
fghij
115.00
46.50
36.75
12.00
42.00
12.00
49.00f
9.00h
ASR
Atomic
Akrave
Clemson
48.40qrs
47.05
rs
44.38
16.00g
19.00
15.00
15.00f
9.00
8.00
16.00hij
27.00
10.25
27.25n
31.75
ij
18.25
6.25mn
9.30
5.75
16.50de
15.00
de
14.75
def
53.75gh
89.75
42.75
5.50kl
7.50
hijk
8.50
efghij
80.00
37.00
10.00
8.00
48.63
21.75lm
43.24
16.50n
spineless
LoS
**
**
**
**
**
**
**
**
**
**
**
**
**
BE
ns
ns
ns
ns
ns
**
ns
ns
**
ns
ns
ns
ns
TCV (%)
11.9
1.1
1.5
6 .1
3.6
2.6
18.0
2.1
16.7
1.2
1.3
0.5
3.8
BCV (%)
1.3
0.3
0.3
1.7
0.5
1.0
2.9
0.2
6.3
0.42
0.3
0.1
1.0
ns kpfkecvgu" pqp" ukipkhkecpeg" cv" vjg" r2027" ngxgn." ," kpfkecvgu" ukipkhkecpeg" cv" vjg" r2027" ngxgn" cpf" ,," kpfkecvgu" jkij" ukipkhkecpeg" cv" r2023" ngxgn0" NqU" ?" ngxgn" qh"
significance, BE = block efficiency, TCV = treatment co-efficient of variation, BCV = block co-efficient of variation and Mean represent average of the individual
characters measured for all accessions under consideration. NV = Northern Version, MPH = Maximum plant height (cm), MNI = Maximum number of internodes, FFN
= First flowering node, NLPP = Number of leaves per plant, ASPD = Average seeds per pod, STB = Stem diameter at base (mm), NPPP = Number of pods per plant,
FPGer = 50% Germination, FFPN = First fruit producing node, FPFl = 50%Flowering, FPFr = 50%Fruiting, TSwt = 1000 seed weight (g), APP = Average pods per plant.
"
PES
SES
NES
RCPB
Cs-Legon
Persistent
Lanceolate
KortebortorASR
Mapelega
Persistent
Lanceolate
Persistent
Triangular
Kpeve
Linear
Yeji-Local
Nonpersistent
Persistent
Amanfrom
Persistent
Lanceolate
Cape
Persistent
Triangular
AsontemASR
Mamolega
Persistent
Triangular
Persistent
Lanceolate
Atomic
Nonpersistent
Lanceolate
More
than 10
More
than 10
More
than 10
From 5
to 7
From 5
to 7
More
than 10
More
than 10
More
than 10
From 5
to 7
From 8
to 10
Inside
only
Inside
only
Inside
only
Inside
only
Inside
only
Both
sides
Inside
only
Inside
only
Both
sides
Inside
only
Triangular
Petal
colour
Cream
Fruit
pubescence
Downy
Cream
Prickly
Cream
Cream
Slightly
rough
Slightly
rough
Prickly
Yellow
Downy
Cream
Cream
Slightly
rough
Slightly
rough
Prickly
Cream
Downy
Cream
Cream
Fruit
colour
Yellowish
green
Green with
red patches
Yellowish
green
Green
PFMS
Erect
Stem
pubescence
Slight
Erect
Slight
Horizontal
Conspicuous
None
(smooth)
None
(smooth)
5 to 10
Pendulous
Slight
5 to 10
Pendulous
Slight
Pendulous
Slight
None
(smooth)
None
(smooth)
5 to 10
None
(smooth)
8 to 10
None
(smooth)
Green with
red patches
Yellowish
green
Green with
red patches
Green
Erect
Conspicuous
Erect
Slight
Red
Pendulous
Conspicuous
Green
Erect
Slight
NRPPD
Fruit
shape
1
7
8
1
7
NES= Number of Epicalyx Segment; RCPB= Red Colouration of Petal Base; PES=Persistence of Epicalyx Segment; SES=Shape of Epicalyx Segment; PFMS=Position
of Fruit on Main Stem; NRPPD=Number of Ridges per Pod.
"
Vcdng"505"*Eqpvf+"
TRAITS AND DESCRIPTION
Accession
PES
SES
NES
RCPB
Asontem-ER
Persistent
Triangular
Juaboso
Persistent
Lanceolate
Debo
Nonpersistent
Persistent
Triangular
Agric short
fruit
Asontem nv
Persistent
Triangular
Inside
only
Inside
only
Inside
only
Inside
only
Both sides
Persistent
Lanceolate
Labadi
Persistent
Lanceolate
Indiana
Persistent
Lanceolate
Asante type II
Persistent
Lanceolate
Legon fingers
Persistent
Lanceolate
More than
10
More
than10
From 7 to
10
More than
10
More than
10
From 7 to
10
More than
10
More than
10
From 8 to
10
More than
10
Volta
Triangular
Petal
colour
Cream
Cream
Cream
Cream
Cream
Both sides
Cream
Inside
only
Both sides
Cream
Inside
only
Inside
only
Cream
Cream
Cream
Fruit
pubescence
Slightly
rough
Slightly
rough
Slightly
rough
Prickly
Slightly
rough
Slightly
rough
Prickly
Slightly
rough
Slightly
rough
Downy
Fruit
colour
Yellowish
green
Green
PFMS
NRPPD
Horizontal
Stem
pubescence
Slight
5 to 10
Fruit
shape
2
Erect
Slight
5 to 10
Green
Pendulous
Slight
5 to 10
10
Green with
red patches
Green
Pendulous
Slight
8 to 10
Pendulous
Conspicuous
5 to 10
12
Green
Erect
Slight
5 to 10
14
Yellowish
green
Yellowish
green
Green
Horizontal
Slight
8 to 10
Horizontal
Glabrous
8 to 10
Erect
Glabrous
5 to 10
Erect
Conspicuous
None
(smooth)
Yellowish
green
NES= Number of Epicalyx Segment; RCPB= Red Colouration of Petal Base; PES=Persistence of Epicalyx Segment; SES=Shape of Epicalyx Segment; PFMS=Position
of Fruit on Main Stem; NRPPD=Number of Ridges per Pod.
"
Vcdng"505"*Eqpvf+
TRAITS AND DESCRIPTION
Accession
PES
Clemson
Non-
Spineless
persistent
Wune mana
Persistent
Agric type I
Kortebortor-
Persistent
Persistent
SES
From 7
Both
to 10
sides
Linear
More
Both
than 10
sides
From 8
Inside
to 10
only
Lanceolate
Lanceolate
Persistent
Triangular
Persistent
Lanceolate
nkuruma
AsontemBAR
DKA
Asontem-
Persistent
Persistent
Lanceolate
Lanceolate
GAR
Akrave
Persistent
RCPB
Linear
BAR
Nkran
NES
Linear
More
Both
than 10
sides
From 8
Inside
to 10
only
More
Inside
than 10
only
More
Both
than 10
sides
More
Inside
than 10
only
From 7
Inside
to 10
only
Petal
Fruit
Fruit
colour
pubescence
colour
PFMS
Stem
NRPPD
Cream
Slightly
Green
Erect
Slight
8 to 10
Green
Erect
Conspicuous
5 to 7
Erect
Conspicuous
5 to 7
Pendulous
Conspicuous
5 to 7
Horizontal
Slight
5 to 7
Erect
Slight
None
pubescence
Fruit
shape
rough
Cream
Slightly
rough
Cream
Cream
Cream
Cream
Cream
Slightly
Yellowish
rough
green
Slightly
Yellowish
rough
green
Slightly
Yellowish
rough
green
Slightly
Yellowish
rough
green
Downy
Green with
(smooth)
Horizontal
Glabrous
8 to 10
Green
Erect
Conspicuous
5 to 7
11
Green with
Pendulous
Glabrous
5 to 10
11
red patches
Cream
Slightly
rough
Cream
Downy
red patches
NES= Number of Epicalyx Segment; RCPB= Red Colouration of Petal Base; PES=Persistence of Epicalyx Segment; SES=Shape of Epicalyx Segment; PFMS=Position
of Fruit on Main Stem; NRPPD=Number of Ridges per Pod.
"
Vcdng"505"*Eqpvf+"
TRAITS AND DESCRIPTION
Accession
Asontem-ER
Juaboso
Debo
PES
Persistent
Persistent
Non-
SES
Triangular
Lanceolate
Triangular
persistent
Volta
Agric short
Persistent
Persistent
Triangular
Triangular
RCPB
More than
Inside
10
only
More
Inside
than10
only
From 7 to
Inside
10
only
More than
Inside
10
only
More than
Both sides
Petal
Fruit
Fruit
colour
pubescence
colour
Cream
Slightly
Yellowish
rough
green
Slightly
Cream
Persistent
Lanceolate
From 7 to
Cream
Indiana
Persistent
Persistent
Lanceolate
Lanceolate
Cream
Persistent
Lanceolate
II
Legon
fingers
Persistent
Lanceolate
NRPPD
pubescence
Fruit
shape
Horizontal
Slight
5 to 10
Green
Erect
Slight
5 to 10
Slightly
Green
Pendulous
Slight
5 to 10
10
Prickly
Green with
Pendulous
Slight
8 to 10
Green
Pendulous
Conspicuous
5 to 10
12
Green
Erect
Slight
5 to 10
14
Yellowish
Horizontal
Slight
8 to 10
Horizontal
Glabrous
8 to 10
red patches
Cream
Slightly
rough
Both sides
Cream
Slightly
rough
More than
Inside
10
only
More than
Both sides
Cream
From 8 to
Inside
10
only
More than
Inside
10
only
Prickly
green
Cream
10
Asante type
Stem
rough
10
Labadi
PFMS
rough
10
fruit
Asontem nv
NES
Cream
Slightly
Yellowish
rough
green
Slightly
Green
Erect
Glabrous
5 to 10
Yellowish
Erect
Conspicuous
None
rough
Cream
Downy
green
"
(smooth)
Vcdng"505"*Eqpvf+"
TRAITS AND DESCRIPTION
Fruit
peduncle
Leaf
colour
Round
Aspect of
seed
surface
Downy
1 to 3cm
Mixed
Reniform
Downy
1 to 3cm
Green with
red veins
Green
Early
Erect
Reniform
Downy
1 to 3cm
Green
Late
Mixed
Round
Glabrous
1 to 3cm
Green
Early
Erect
Round
Glabrous
Green
Strong
Late
Medium
Round
Downy
More
than 3cm
1 to 3cm
Mixed
Stocky
Medium
Midseason
Erect
Round
Glabrous
1 to 3cm
Green
10
Curved
Medium
Early
Procumbent
Reniform
Glabrous
Green
Medium
11
Mixed axes
Midseason
Procumbent
Reniform
Downy
Mixed
Straight
Orthotropic
stem only
Medium
More
than 3cm
1 to 3cm
Midseason
Medium
Round
Glabrous
Accession
Pod
length
Pod
diameter
Leaf
shape
Pod axis
type
Branching
type
PMS 1-9
General
plant type
Seed
shape
Asontem nv.
Very
short
Very
long
Very
long
Mixed
Mixed axes
Medium
Midseason
Medium
Big
Mixed axes
Medium
Late
Medium
10
Straight
Orthotropic
stem only
Asante type II
Medium
Mixed
10
Stocky
Legon fingers
Short
Medium
Stocky
Orthotropic
stem only
Medium
Akrave
Short
Big
Curved
Atomic
Short
Small
Clemson
spineless
Wune mana
Long
Medium
Medium
Agric type I
Medium
Labadi
Indiana
"
More
than 3cm
Green with
red veins
Green with
red veins
Vcdng"505"*Eqpvf+"
TRAITS AND DESCRIPTION
Accession
Pod length
Pod
Leaf
Pod axis
Branching
diameter
shape
type
type
PMS 1-9
General
Seed shape
plant type
Aspect of
Fruit
seed surface
peduncle
Leaf colour
Asontem-BAR
Medium
Mixed
Stocky
Mixed
Midseason
Erect
Round
Downy
1 to 3cm
Green with
DKA
Medium
Mixed
Mixed
Strong
Very late
Mixed
Reniform
Downy
More than
Green with
3cm
red veins
Asontem-GAR
Medium
Big
Stocky
Mixed
Late
Erect
Reniform
Glabrous
More than
Mixed
red veins
axes
3cm
Kortebortor-
Medium
Medium
Straight
Strong
Very late
Erect
Reniform
Downy
More than
Green
3cm
BAR
Nkran nkuruma
Mixed
Big
Mixed
Medium
Very late
Mixed
Reniform
Glabrous
Debo
Medium
Medium
Stocky
Strong
Late
Medium
Reniform
Downy
1 to 3cm
Volta
Long
Medium
Green
Curved
Medium
Early
Erect
Round
Glabrous
1 to 3cm
Green
Medium
Medium
Straight
Medium
Early
Medium
Reniform
Glabrous
1 to 3cm
Green
Juaboso
Medium
Medium
Mixed
Medium
Midseason
Erect
Reniform
Glabrous
1 to 3cm
Green
axes
axes
"
More than
Green with
3cm
red veins
"
"
Fifty percent (50%) flowering was positively correlated with traits such as
50% fruiting, maximum number of internode, first-flowering node, number
of leaves per plant, stem diameter at the base, number of pods per plant but
negatively correlated with all the other traits. The association of 50%
flowering with all other traits whether negative or positive were not
statistically significant except with 50% fruiting.
On the other hand, 50% fruiting was strongly correlated with number of
leaves per plant and stem diameter at the base as well as positively correlated
with other traits such as first-flowering node, average number of seeds per
pod, number of pods per plant and first fruit-producing node. Nevertheless, it
was negatively correlated with characters such as the maximum plant height,
maximum number of internodes, 1000-seed weight and average number of
pods per plant. Maximum plant height was correlated positively with all
other traits except 50% germination, 50% fruiting and 50% flowering.
"
"
"
"
FPGer
FPFl
FPFr
MPH
MNI
FFN
NLPP
TSwt
ASPPD
STB
APP
NPP
FPGer
FPFl
-0.24
FPFr
-0.19
0.95**
MPH
-0.03
-0.05
-0.03
MNI
-0.08
0.05
-0.03
0.42
FFN
-0.13
0.05
0.01
0.62*
0.53*
NLPP
-0.22
0.43
0.57*
0.12
0.12
0.15
TSwt
-0.07
-0.17
-0.13
0.39
0.18
0.30
0.29
ASPPD
-0.03
-0.13
0.08
0.34
0.14
0.36
0.57*
0.30
STB
-0.03
0.44
0.50*
0.36
0.33
0.37
0.46
0.16
0.24
APP
0.11
-0.10
-0.05
0.45
0.42
0.71**
0.26
0.21
0.60*
NPPP
-0.28
0.14
0.14
0.44
0.47
0.68**
0.41
0.24
0.46
0.29
0.64*
FFPN
-0.02
-0.05
0.01
0.14
0.27
0.20
0.00
-0.06
0.29
0.14
0.10
*Significant at P = 0.5
"
0.22
0.30
FFPN
The five PCs accounted for about 78.51% of total variance with the first
principal component (PC1) recording the highest (32.44%). The second, third,
fourth and fifth principal components (PC2, PC3, PC4 and PC5) accounted for
19.78%, 9.68%, 8.45% and 8.15% respectively. The Eigen values shows the
relative discriminating power of the principal axes with (4.22) for axis 1 and as
low as (1.06) for axis 5. PC1, which accounted for the highest proportion
(32.44%) of total variation mostly correlated with number of pods per plant, first
flowering node, average pods per plant, maximum plant height, average seeds
per pod, maximum number of internode, stem diameter at the base, number of
leaves per plant and 1000-seed weight.
Average pods per plant, maximum plant height, 50% germination, first
flowering node and 1000-seed weight, 50% fruiting, 50% flowering; number of
leaves per plant, stem diameter at the base and maximum number of internodes
were mostly correlated with PC2.
The third principal component (PC3) was positively correlated with maximum
number of internodes, first fruit-producing node, first flowering node, 50%
flowering but negatively correlated with number of leaves per plant, 1000-seed
weight, average seeds per pods, average pods per plant. First fruit-producing
"
node, average seeds per pods, 50% germination, and 1000-seed weight made
substantial contribution to PC4. Finally, 50% germination, first fruit-producing
node and stem diameter at the base substantially contributed to PC5.
Table 3.5: Principal components analysis showing the contribution (factor scores) of 13
quantitative characters among the 29 accessions of Okra, Eigen values and Percentage
total variance accounted for by five principal components *.
CHARACTER
PC1
PC2
PC3
PC4
-0.44768
-0.41751
PC5
-0.02082
-0.08673
1000-Seed weight
0.203819*
0.153724*
50% Flowering
0.111969
-0.56337
0.184005*
-0.07344
50% Fruiting
0.14188
-0.57211
0.02661
0.07007
-0.10149
50% Germination
-0.09528
0.17753*
-0.05589
0.36608*
-0.78663
0.35933*
0.21348*
-0.05857
0.14673
-0.14057
0.318568*
0.09009
-0.43818
0.41697*
0.11012
0.389668*
0.16413*
0.22046*
-0.16218
-0.04067
0.14729
0.08145
0.34392*
0.59379*
0.27169*
0.29276*
0.11885
0.41954*
-0.14524
-0.04057
0.32356*
0.1805*
0.08216
-0.26811
-0.20185
0.28419*
-0.3141
-0.44695
0.11124
0.07890
0.39654*
0.0471
0.09077
0.03341
0.26771*
0.29013*
-0.2554
0.09383
-0.02465
-0.36787
Eigen value
4.22
2.57
1.26
1.09
1.06
% Variance
32.44
19.78
9.68
8.45
8.15
Cumulative % Variance
32.44
52.23
61.90
70.35
78.51
* Values bolded and asterisked made substantial contribution to total variance in the respective axes. Maximum
and least discriminating power (eigen value), maximum and least percentage variance and maximum cumulative
percentage variance values are bolded.
"
3.6 DISCUSSION
3.6.1 Variation in Quantitative and Qualitative Traits
The 29 accessions of okra exhibited significant variation in all but two
quantitative traits studied. Treatment coefficients of variation were low for all
traits. Block coefficients of variation were extremely low but high for stem
diameter at the base and first fruit-producing node, implying that results
obtained are reliable and repeatable over replications. Number of pods per plant,
first fruit-producing node and maximum plant height recorded the highest
treatment coefficients of variation but these were within ranges obtained by
Gomez and Gomez (1984) and less than what was obtained by Ogunbayo
(2005). On the other hand, all the accessions showed similarity in some
qualitative traits and variability in other qualitative traits. The highly significant
differences observed for most characters investigated are an indication of their
genetic diversity (Amoatey, 1987).
"
"
"
L.), rapeseed (Brassica napus L.), Asian okra (Abelmoschus esculentus L.) and
mandarin (Citrus spp) respectively.
Again, height of plants in this study varied significantly among the okra
accessions studied (Table 3.2). Nkran Nkuruma recorded a mean height of
170.78cm followed by Asontem-BAR, Asante type II, Asontem-GAR, YejiLocal, and Cape recording mean maximum plant height of 128.53cm, 116.33cm,
113.40cm, 103.95cm, and 102.53cm respectively. On the other hand, the
accessions; Kpeve, Kortebortor-ASR, Atomic, Akrave and Clemson spineless
recorded shortest mean plant height of 56.68cm, 48.50cm, 48.40cm, 47.05cm
and 44.38cm respectively. Height at flowering and fruiting (final height) are of
particular interest for breeding programmes because tall, thin stems increase rate
of lodging near harvesting time (Doku, 2011) and this could culminate in loss of
dry matter and a subsequent decrease in fruit yield. This is in consonance with
reports by Doku (2011), Akinyele and Oseikita (2006) and Myanmar (1995)
who worked on rice and okra respectively.
"
"
with end of seed formation. Therefore, the longer the duration of the vegetative
stage, the longer the life cycle of the plant and vice versa (Doku, 2011).
Results obtained in this study show that the okra accessions exhibited varying
degrees of fruit pubescence spanning from prickly, slightly rough (smooth),
downy to little hairs on fruits but with the majority having smooth fruits. This
result contrasts those of Bish et al. (1995) and Thomas (1991) who found downy
type of fruit pubescence to be the most pronounced, followed by slightly rough
while prickly fruits was the least in the okra accessions they studied. This could
dg"cp"kpfkecvkqp"qh"rtghgtgpeg"qh"Ijcpckcp"hctogtu"hqt"uoqqvj"htwkv"v{rgu"cpf"
thus, discarded the hairy types (Oppong-Sekyere, 2011).
Petal colour was either cream or yellow with golden yellow flowers (cream)
recording the highest. This is contrary to observation made by Myanmar (1995)
who found petal colour to be 100% yellow for all 40 okra accessions examined.
Akinyele and Akinlosotu (1991) also found similar results in their okra research.
"
Variations observed in this study were conspicuous for position of fruit on main
stem that was erect, horizontal or pendulous. Most of the accessions had their
fruits with erect position on main stem as against pendulous and horizontal
positioning of fruits on main stem of the accessions. This is because different
genotypes have the tendency of exhibiting different growth habits, whether as a
result of selection or a natural adaption mechanism. This is similar to an
observation made by Hanson (2005) working on tomato. Yellowish green was
the most predominant fruit colour while green and green with red patches were
least observed among the accessions. Similar observation was found with leaf
colour. These results are similar to those found by Oppong-Sekyere (2011) and
Myanmar (1995) in okra.
"
The first sub-cluster and last two sub-sub clusters exhibited higher inter-cluster
distance and accordingly could be utilised for obtaining heterobeltiosis. The
accessions Cs-Legon, Mamolega, and Wune mana, which appear to be the most
diverse, may be potentially useful as sources of variable characters in okra
improvement programme, as affirmed by Irwin et al. (1998) that closely related
accessions are normally located within 8090% similarity. Crosses between
accessions with similarity indices of 80100% may not be desirable; but the
potential for successful crossing of unrelated varieties may produce an array of
genotypes from which useful agronomic types may be selected (Gulsen et al.,
2007).
"
Cs-Legon was the most diverse and isolated in a cluster. This may be explained
by its unique characteristics of persistent epicalyx segment, lanceolate shape of
epicalyx segment, creamy petal colour, downy fruit pubescence, yellowish green
fruit colour, erect position of pods on main stem, slight stem pubescence,
smooth pods, medium pod length and diameter, stocky pod axis, round seed
shape, green leaf colour and prolific yielding potential.
Mamolega and Wune mana were placed in sub-cluster five. This coincides with
the geographical origin of the accessions. The two accessions were collected
from the same geographical location (Upper East Region) and possess similar
morpho-characteristics (Belete, 2011), as a reflection of adaptation to similar
environmental conditions or related ancestry.
Asontem-ER and Volta had the highest genetic similarity index (GS) of 94.1%,
followed by Asante type II and Agric type I (91.5%), Cape and Asontem-ASR
(91%), Kpeve and Debo (90.6%), and Kortebortor-ASR and Asontem-BAR
(90.4%) all separated in cluster two except Kpeve and Debo which were
separated in cluster one into a different sub-cluster. There were however, no
duplicates among the accessions collected. For accessions to be taken as
genetically identical, their genetic similarity index (GS) should be equal to or
greater than 95% (Andersson et al., 2007). In this study, the maximum genetic
similarity was 94.1% recorded between Asontem-ER and Volta (as the closest
pair of accession).
The large size of the accessions from a wide range of geographical areas is very
essential for genetic distance estimation (Nei, 1978). Broad range of genetic
similarity indices and clustering of accessions provides useful variability in okra
"
"
The output of the PCA revealed that different characters contributed differently
to the total variation. The mean contributions of maximum plant height, 50%
germination, average seeds per pod, average pods per plant, first flowering node,
first fruit-producing node, maximum number of internodes, number of leaves
per plant, number of pods per plant were relatively high in the principal axes.
This observation confirmed the individual contributions of these traits to the
variations observed in the 29 okra accessions as well as seed yield in all the
accessions. This agrees with the report of Ogunbodede (1997), Ariyo, and
Odulaja (1991). It further suggests that selection for seed yield must take into
consideration these traits. This corroborates the report of Aremu et al. (2007)
working with cowpea and Olika et al. (2011) working with coffee.
The first four principal axes accounted for over 70% of total variation among the
13 characters describing the accessions. These characters can be employed in
differentiating okra accessions (Nwangburuka et al., 2011; Clifford and
Stephenson, 1975). The total contribution of the five principal component axes
of this study was higher (78.51%) than observations made by other workers
(Nwangburuka et al., 2011; Moukoumbi et al., 2011; Campos et al., 2005;
Ogunbayo et al., 2005) where the principal component axes contributed 64.32%,
"
66.37%, 76.62% and 64.5% variation, respectively. In this present study, all the
eigen values were lower than what was observed by Nwangburuka et al. (2011).
First fruit-producing node and number of leaves per plant were found to have
contributed positively and significantly to total genetic variance in this study,
confirming a similar observation by Nwangburuka et al. (2011). The factor
scores of the 12 characters imply that, high priorities should be given to these
genetic traits, if selection is to be made for a future breeding programme.
3.7 CONCLUSION
The following conclusions can be drawn from the current study:
I.
II.
III.
Cs-Legon was the most diverse with outstanding traits such as smooth
pod shape, early fruiting, uniform-sized, medium height, prolific fruiting
potential, straight and stocky stem, and smooth pod shape (no spines).
Mamolega and Wune mana were also identified as most diverse and distantly
related accessions and could be utilised as parent stocks to introgress the
desirable traits inherent in them into the exportable cultivar Indiana, and as
sources of variability for future breeding work.
"
IV.
"
"
"
REFERENCES
Adeniji, O. T. (2003). Inheritance studies in West African okra (A. caillei). M.
Agric. Thesis. University of Agriculture, Abeokuta, Nigeria, 98p.
Adeniji, O.T., Aremu, C.O. (2007). Interrelationships among characters and path
analysis for pod yield components in West African okra (Abelmoschus caillei A,
Chev) Stevels). Journal of Agronomy, 6(1):162-166.
"
"
Andersson, M.S., Schultze-Kraft, R., Peters, M., Duque, M.C. and Gallego, G.
(2007). Extent and structure of genetic diversity in a collection of the tropical
multipurpose shrub legume Cratylia argentea (Desv.) O. Kuntze as revealed by
RAPD markers. Electronic Journal of Biotechnology, 10 (3): 1-9.
Aremu, C.O., Adebayo, M.A., Ariyo, O.J., Adewale, B.B. (2007). Classification
of Genetic diversity and choice of parents for hybridisation in cowpea Vigna
unguiculata (L.) Walp for humid savanna ecology. Afr. J. Biotechnol., 6(20):
2333-2339.
"
"
C{vc." 0" cpf" Mpce." I0" *422;+0" Igpgvke" xctkcdknkv{" cpf" cuuqekcvkqp"uvwfkgu"qh"
some quantitative characters in winter rapeseed (Brassica napus L.). Afri. J.
Biotechnol., 15: 3547-3554.
"
Belete, Y.S. (2011). Genetic variability, correlation and path analysis studies in
Ethiopian mustard (Brassica carinnata A. Brun) Genotypes. International
Journal of Plant Breeding and Genetics 5 (4): 328-338.
"
Bisht I.S. and Bhat, K.V. (2006). Genetic resources, chromosome engineering
and crop improvement series, Edited by Ram J. Singh, 3:148-162.
Campos, E.T., Espinosa, M.A.G., Warburton, M.L. and Monter, A.V. (2005).
Characterisation of madrin (Citrus spp) using morphological and AFLP markers.
Interciencia 30 (11): 1-14.
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"
"
"
"
Gomez, K. A. and Gomez, A.A. (1984). Single factor experiment. In: Statiscal
Procedures for Agricultural Research. (IRRI, ed.), 2nd Ed, John Wiley and Sons,
New York, U.S.A, pp.16-17.
Gulsen, O., Karagul, S., Abak, K. (2007). Diversity and relationships among
Turkish germplasm by SRAP and Phenotypic marker polymorphism. Biologia,
Bratislava, 62(1): 41-45.
Hamon, S., Nairot, M. (1991). Some proposed procedures for obtaining a core
collection using quantitative plant characterisation. International Workshop on
okra genetic resources held at NBPGR. International Crop Network Series No.
5: 89-94.
"
"
Hazra, P. and Basu, D. (2000). Genetic variability, correlation and path analysis
in okra. Ann Agril Res. 21(3): 452-453.
Hien, N. L., Sarhadi, W. A., Oikawa, Y. and Hirata, Yutaka. (2007). Genetic
diversity of morphological responses and the relationships among Asia aromatic
rice (Oryza sativa L.) cultivars. Tropics, 16 (4): 333-355.
"
"
Kaul, T., Lal, G. and Peter, K. V. (1978). Correlation and path coefficient
analysis of components of earliness, pod yield and seed yield in okra. Indian J.
Agric. Sc. 48(8): 459-63.
"
Martin, F.W., Rhodes, A. M., Ortiz, M. and Diaz, F. (1981). Variation in okra.
Euphytica, 30: 697-705.
"
Mehta, D.R., Dhaduk, L.K. and Patel, K.D. (2006). Genetic variability,
correlation and path analysis studies in okra {Abelmoschus esculentus (L.)
Moench}. Agric. Sci. Digest, 26 (1): 15 18.
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Oqwmqwodk."[0"F0."Uk."O0."Xqfqwjg."T0."Pftk."D0."Vqwnqw."D0."Qiwpdc{q."U0"
A. and Ahanchede, A. (2011). Assessing phenotypic diversity of interspecific
rice varieties using agro-morphological characterisation. Journal of Plant
Breeding and Crop Science. 3(5): 74-86.
"
"
"
Olika, K., Sentayehu, A., Taye, K., Weyessa, G. (2011). Genetic diversity
analysis in Limmu coffee (Coffea Arabica L.) collection using quantitative traits
in Ethiopia. Int. J. Agric. Res. 6(6): 470-481.
Patil, Y.B., Madalageri, B.B., Biradar, B.D. and Hosamani, R.M. (1996).
Variability studies in okra (Abelmoschus esculentus (L.) Moench). Karnataka J.
Agric. Sci., 9: 289-293.
Payne, R.W., Harding, S.A., Murray, D.A., Soutar, D.M., Baird, D.B., Welham,
S.J., Kane, A.F., Gilmour, A.R., Thompson, R., Webster, R., Tunnicliffe, G. W.
(2007). Genstat Statistical Programme, Ninth Edition. Lawes Agricultural Trust
(Rothamsted
Experimental
Station).,
vers.9.2.0.152.PC/Windows,
VSN
Rubenstein, K. and Heisey, P. (2003). Plant Genetic Resources: New Rules for
International Exchange, Bioversity's Regional Office for the Americas, IPGRI.
Villa Gualino, Turin, Italy. 63pp.
Singh, H. B., Swarup, V., Singh, B. (1974). Three decades of vegetable research
in India. ICAR Technical Bulletin, New Delhi. 128 pp.
Singh, K.B. and Singh, H.N. (1977). Path coefficient analysis for yield in okra.
Indian J. Agric. Sci. 49: 244-246.
Statgraphics. (2010). Statgraphics Centurion XVI, version 16.1.11, Windowsbased statistical software, (32-bit) 2010 Statpoint Technologies, Inc.
Multilingual, USA.
"
Staub, J.E., Serquen, J.C. and Gupta, M. (1996). Selection for multiple lateral
determinate cucumber genotypes. Cucurbit Gen. Coop. Rpt. 18: 5-6.
Steel, R.G., Torrie, R.H. (1984). Principles and procedure of statistics. 2nd
edition McGraw-Hill, Inc. New York. 87pp.
Vogel, J. M., Rafalski, A., Powell, W., Morgante, M., Andre, C., Hanafey, M.
and Tingey, S. V. (1996). Application of genetic diagnostics to plant genome
analysis and plant breeding. Hort Sci., 31: 165-167.
"
CHAPTER FOUR
4.1 INTRODUCTION"
The most significant breakthrough in agricultural biotechnology is the
understanding of the structure of genomes and the genetic mechanisms behind
economically important traits. Plant genomics also known as plant molecular
biology is now able to provide information on the identity, location, impact and
functions of genes controlling traits, which can be identified, catalogued and
mapped as single gene markers in countless species of higher plants (Jonah et
al., 2011). Molecular markers are identifiable DNA sequences, found at specific
loci (locations) within the genome and associated with inheritance of a trait or
linked gene (FAO, 2004). Thottappilly et al. (2000) defined molecular markers
as naturally occurring polymorphism, which include proteins and nucleic acids
that are detectably different. Thus, molecular DNA markers are individual genes
flanking within a defined close interval in the genome of an organism.
frequently associated with specific gene and cevu" cu" c" ukip" rquvu" vq" wughwn"
genes and such markers when very tightly linked to genes of interest, can be
exploited to select indirectly for desirable alleles and this represents the simplest
form of marker- assisted selection (MAS) (Hoisington et al., 2002).
(Lindroos
et
amplified
fragment
length
polymorphisms (AFLP) (Karp et al., 1997; Vos et al., 1995). The different
genetic markers vary with respect to important features such as genomic
"
"
"
genetic diversity and polymorphism within the cultivated species of the genus
Abelmoschus. West Africa alone holds 1,769 of the 2,283 accessions reported
worldwide. Variability is more pronounced particularly in plant height, days to
flowering and fruiting, tolerance to the yellow vein mosaic virus, yielding
potential and a host of other fruit characteristics among okra germplasm (Gulsen
et al., 2007; Ariyo and Odulaja, 1991). On the contrary, there is little research
investment towards improving the crop in the sub-region though being the
second largest producer globally.
"
Okra is put to diverse uses by different end users. This is because every part of
this crop seems to offer some useful purpose or the other. Varieties are needed
as sources of protein, fibre, biomass, oil, mucilage (type and yield), colour,
medicinal and ornamental use (National Academies Press, 2006). Traditional
farmers often engage in selection for different purposes and eventually mixed
them up. Okra in the hands of these traditional farmers diversifies into vast
number of cultivars (landraces) adapted to various environments (De Lannoy,
2001). However, with the emerging status of okra as an export crop, there is
therefore the need for standardisation of traits inherent in these landraces to meet
specific end uses and markets.
Although, there has been considerable selection and breeding of okra, it has
been limited to production of immature pods. The rest of the fantastic genetic
diversity within this species is untapped, or even unexplored. Molecular
characterisation is the ultimate tool to exploit the genetic variability (intra and
interspecific variations) in this crop from different phyto-geographic areas; this
will open the way for improving the compositional value of the crop, improving
"
"
ISSR markers are quick and easy to handle, but the longer length of their
primers make them seem to have the reproducibility of SSR markers. ISSR
primers contain sequences complementary to SSR motif with 1-3 base anchors
cv" gkvjgt" vjg" 5" qt" 7" gpf." dcug" rqukvkqp" ykvjkp" vjg" cpejqt" cpf" oc{" jcxg" cp{"
nucleotide other than that needed to continue the repeat sequence ensuring that
the primer binds exclusively to one end of the targeted SSR and not in the
middle. Numerous amplification products are generated by ISSRs, which
"
"
Gcej"ucorng"ycu"vjgp"ycujgf"cickp"ykvj"822" n"ejnqtqhqto"CKC"cpf"urkppgf"cv"
1300 rpm for 10 minutes. Sample supernatant of each was then pipetted into a
new 1.5ml microfuge tube. The DNA of each sample was then precipitated with
vjg"wug"qh"822 n"kuqrtqrcpqn"cpf"gcej"oketqhwig"vwdg"ycu"vjgp"igpvn{"kpxgtvgf0"
The samples were then stored at -20C for 30 minutes. Afterwards, each sample
was then spinned at 1300 rpm for 15 minutes. The supernatant of each sample
was carefully eluted out to leave the DNA pellet, and allowed to dry. The pellet
was reuwurgpfgf"kp"322 n"VG"buffer and stored at -20 C for later use.
4.2.4 Amplification
The amplification was done with thermal cycler from BIO-RAD. The
temperature cycles were, a first denaturation step of 5 min at 94C, followed by
35 cycles of 94C for 30s, 55C for 30s and 72C for 1min followed by a final
extension at 72C for 7minutes. The complete reactions were then held at 4C
until electrophoresis. The amplicons were verified for amplification by
electrophoresis on 1.5% agarose gel.
"
4.2.6 Electrophoresis
The PAGE was warmed in the electrophoresis tank at 250volts for 15minutes to
ensure the removal of any excessive sanv" kp" vjg" ign0" Chvgtyctfu." 407 n" qh" vjg"
cornkeqpu" qh" vjg" xctkqwu" ceeguukqpu" ygtg" vjqtqwijn{" okzgf" ykvj" 4 n" nqcfkpi"
dye and loaded into the wells of the gel. 50bp DNA ladder (marker) was also
loaded in one well to estimate the molecular size of the amplicons. It was then
ran in 1X TBE for 1hour; first 40mins at 100volts to remove the running
"
amplicons from the wells and set it running and the remaining 80minutes at
140volts to enable amplicons move fast and prevent the fading of bands.
Afterwards, it was then stained in 400ml of 0.05% silver stain via orbital shaker
for 1hour and then photographed using BioDoc-it gel documentation system
(http.www.molecularinfo.com/MTM/C/C1/C1-1.html).
"
"
4.4 DISCUSSION
4.4.1 Cluster Analysis
Assessment of the extent of relatedness among potential parents forms the
foundation for selection of genetically distant and diverse parents in any
breeding programme. The accessions Asante type II, Yeji-Local, Atomic and
Indiana were contained in cluster entry 1 whilst the remaining 25 accession were
all grouped into cluster entry 2. This could be explained by their outstanding
characteristics (relatively early fruiting, uniform in shape, moderate level of
mucilage, medium in height, glabrous stem, and moderate length of peduncle),
as these accessions were collected from different geographical areas. Asante
type II from the Brong-Ahafo region and Yeji-Local (forest savanna belt),
Atomic from the greater Accra region (coastal savanna zone) and Indiana from
India (template climatic region).
Indiana however was the most diverse with a genetic similarity index of 96.1%.
However, the wide variation between Indiana and the rest of the populations
indicates the usefulness of each population, particularly Asante type II, YejiLocal, Atomic populations which were most diverse, as a valuable genetic
resource for selection of superior genotypes. Indiana is an exotic cultivar and is
not well adapted to the local conditions but possess appreciable desired qualities
(slim and narrow in shape and size, smooth, yellowish green in colour, early
fruiting, short peduncle, moderate level of mucilage and less bulky in weight)
for the export market. Asante types II, Yeji-Local, Atomic are all indigenous
cultivars, well adapted to local conditions and very hardy to both biotic and
abiotic stresses yet non-uniform in shape and size. The desirable traits in Indiana
"
could be introgressed into any of these three accessions to improve them for
both export market and local consumption.
"4.4.2
ISSR Primers
The genetic relationships among individual accessions or groups are further
elucidated by the dendrogram generated via the ISSR primers. Results obtained
from the dendrogram reveal two major clusters at a genetic or similarity distance
of 96% for all the accessions studied. All 25 accessions in group II could be
considered as possible duplicates with genetic similarity index of 100%
(Andersson et al., 2007). This suggests that these accessions are genetically
closely related or they have a common ancestry descent (Kalivas et al., 2011).
This contrasts with findings by Pasqualone et al. (2012), Yao et al. (2008),
Nwangburuka et al. (2011), Aladele et al. (2008), Aladele (2005) who concluded
that there is high genetic diversity among Glycine max, Glycyrrhiza uralensis, A.
esculentus and A. caillei species. Few of the ISSR primers however, were able
to amplify more than one band per genotype; hence, less residual heterogeneity
could be suspected within the lines (Ogunbayo et al., 2005).
The low level of polymorphism detected with the six ISSR primers among the
accessions could be attributed to the dominant nature and low level of locus
specificity associated with ISSR primers. ISSRs are dominant primers and hence
almost unlikely to detect specific loci within genomes, thus unable to detect
heterozygous alleles within genomes of organisms.
"
In addition, two regions, ITS and trnL regions of two accessions (Akrave and
Atomic) were sequenced. The sequenced ITS regions were chaotic and
multicopies indications were found, demonstrating a multiple origin of the
genomic DNA of the individual accession (hybrid). Therefore the trnL intron
(maternal DNA), which is more conserved than the ITS, is a non-functional unit
(intron) of the gene. This indicates that, they are the same, which means that
these two okra accessions originated from the same maternal line (no different
maternal lines) or common source.
4.5 CONCLUSIONS
The use of the six ISSR primers yielded low level of polymorphism among the
29 accessions of okra.
a)
b)
Indiana was the most distantly distinct accession and stand as a unique
"
c)
genotypes
d)
Akrave and Atomic were identified to have originated from the same
ancestry descent.
4.6 RECOMMENDATIONS
For a co-dominant marker to be employed in any characterisation work, the
sequence of the genome of the plant must be deciphered and known. To the best
of my knowledge, the genome of okra has not yet been sequenced hence no SSR
primers are available now to conduct a comprehensive and thorough genetic
diversity study at the gene level. Sequencing calls for huge financial and
infrastructural investments, therefore I recommend the following;
a)
qualities of Indiana, Yeji-Local, Asante type II and Atomic into landraces that
are well adapted to the local agro ecological conditions but lack these qualities
(narrow shape and size, smooth pod pubescence, yellowish green in colour,
early fruiting, short peduncle, moderate level of mucilage, persistent epicalyx
segments, medium pod length and less bulky in weight).
"
"
"
c)
In addition, collections was done from eight regions of Ghana, this should
"
"
"
"
REFERENCES
Akagi, H., Yokozeki, Y., Inakagi, A. (1997). Highly polymorphic microsatellites
of rice consist of AT repeats, and a classification of closely related cultivars with
these microsatellites loci. Theor.Appl.Genet, 94: 61-67.
Andersson, M.S., Schultze-Kraft, R., Peters, M., Duque, M.C. and Gallego, G.
(2007). Extent and structure of genetic diversity in a collection of the tropical
multipurpose shrub legume Cratylia argentea (Desv.) O. Kuntze as revealed by
RAPD markers. Electronic. J. Biotechnology, 10 (3): 1-9.
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Bisht, I.S, Mahajan, R.K., Rana, R.S. (1995). Genetic diversity in South Asia
okra (A. esculentus) germplasm collection. Ann. Appl. Biol., 126: 539-550.
Botstein, D., White, R.L., Skolnick, M., Davis, R.W. (1980). Construction of a
genetic linkage map in man using restriction fragment length polymorphisms.
Am. J. Hum Genet, 32: 314-333.
"
Choi, I.Y., Hyten, D.L., Matukumalli, L.K., Song, Q., Chaky, J.M., Quigley,
C.V., Chase, K., Lark, K.G., Reiter, R.S. and Yoon, M.S. (2007). A soybean
transcript
map:
Gene
distribution,
haplotype
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and
single-nucleotide
Doyle, J.J., Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus,
12: 1315.
Ferriol, M., Pico, B., Nuez, F. (2003). Genetic diversity of germplasm collection
of Cucubita pepo using SRAP and AFLP markers. Theor.Appl.Genet, 107: 271282.
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"
"
Gulsen, O., Karagul, S., Abak, K. (2007). Diversity and relationships among
Turkish okra germplasm by SRAP and phenotypic marker polymorphism.
Biologia Bratislava, 62: 41- 45.
Gupta, P.K., Varshney, R.K., Sharma, P.C. and Ramesh, B. (1999). Molecular
markers and their application in wheat breeding. Plant Breed. 118: 369390.
Hoisington, D., Bohorova, N., Fennell, S., Khairallah, M., Pellegrineschi, A. and
Ribaut, J.M. (2002). The application of biotechnology in wheat improvement
and production. Curtics, B.C. Rajaram S. and Gomez, H. (eds). FAO, pp. 9-12.
http.www.molecularinfo.com/MTM/C/C1/C1-1.html.
Jackson, M.B. and Ram, P.C. (2003). Physiological and molecular basis of
susceptibility and tolerance of rice plants to complete submergence. Ann. Bot,
91: 227-247.
Jonah, P.M., Bello, L. L., Lucky, O., Midau, A., Moruppa, S. M. (2011). The
Importance of Molecular Markers in Plant Breeding Programmes. Global
Journal of Science Frontier Research, 11 (5):1-9.
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Kalivas, A., Xanthopoulos, F., Kehagia, O., and Tsaftaris, A.S. (2011).
Agronomic characterization, genetic diversity and association analysis of cotton
cultivars using simple sequence repeat molecular markers. Genetics and
Molecular Research, 10 (1): 208-217.
Karp, A., Kresovich, S., Bhat, K.V., Ayand, W. G. and Hodgkin, T. (1997).
Molecular tools in plant genetic resources conservation: a guide to the
technologies, IPGRI Tech bull. pp. 2.
Kojima, L., Nagaoka, T., Noda, K. (1998). Genetic linkage map of ISSR and
RAPD markers in einkorn wheat in relation to that of RFLP markers.
Theor.Appl.Genet, 96: 37-47.
Kruglyak, L. and Nickerson, D.A. (2001). Variation is the spice of life. Nat.
Genet., 27: 234236.
Kumar, S., Dagnoko, S., Haougui, A., Ratnadass, A., Pasternak, D. and
Kouame, C. (2010). Okra (Abelmoschus spp.) in West and Central Africa:
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Lindroos, K., Sigurdsson, S., Johansson, K., Ronnblom, L. and Syvanen, A.C.
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Nasu, S., Suzuki, J., Ohta, R., Hasegawa, K., Yui, R., Kitazawa, N., Monna, L.
and Minobe, Y. (2002). Search for and analysis of single nucleotide
polymorphisms (SNPs) in rice (Oryza sativa, Oryza rufipogon) and
establishment of SNP markers. DNA Res., 9:163171.
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National Academies Press. (2006). Lost Crops of Africa Volume II: Vegetables.
pp. 287-301. www.nap.edu/catalog/11763.html.
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Schoen, D.J. and Brown, A.H.D. (1995). Maximizing genetic diversity in core
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Hintum, T.J.L. and Morales. E.A.V. (eds), Core Collections of Plant Genetic
Resources. Chichester: John Wiley, pp.5576.
Thottappilly, G., Magonouna, H.D., Omitogun, O.G. (2000). The use of DNA
markers for rapid improvement of crops in Africa. African Crop Science
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wild bean core collection. Crop Sci, 41:1-8.
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Tsumura, Y., Ohba, K. and Strauss, S.H. (1996). Diversity and inheritance of
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"
van Toai, T.T., St. Martin, S.K., Chase, K. (2001). Identification of a QTL
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Frijters, A., Pot, J., Peleman, J. and Kuiper, M. (1995). AFLP: A new technique
for DNA fingerprinting. Nucleic Acids Res., 23: 44074414.
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(1990). DNA polymorphisms amplified by arbitrary primers are useful as
genetic markers. Nucleic Acids Res., 18: 65316535.
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screening and preliminary evaluation of genetic diversity in wild populations of
Glycyrrhiza uralensis, Biologia Plantarum, 52 (1): 117-120.
"
"
"
Yeboah, M. A., Xuehao, C., Feng, R.C., Liang, G. and Gu, M. (2007). A genetic
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SRAP, and RAPD markers to assess genetic diversity in Turkish melons.
Scientia Horticulturae, 130: 349353.
sequence
repeat
(SSR)-anchored
"
"
polymerase
chain
reaction
"
"
CHAPTER FIVE
"
5.1 INTRODUCTION
Antioxidants are powerful free radical scavengers in the body, while free
radicals are highly reactive chemical substances such as peroxide, hydroxyl
radical and singlet oxygen that travel around in the body and cause damage to
body cells (Alia et al., 2003). Many human degenerative diseases have been
ascribed to free radical damages in the body and numerous studies have been
undertaken on how to delay or prevent onset of these diseases (Oboh, 2005).
Radicals are chemical species with one or more unpaired electrons, and free
radicals are radicals that have moved out of the immediate molecular
environment of their generation. There are several endogenous sources of
oxidants in the body: reduction of molecular oxygen in mitochondria during
cellular respiration takes place in sequential steps (Halliwell et al., 1995),
yielding the radical by-products superoxide (O2), hydroxyl (OH ), and hydrogen
peroxide (H2O2); degradation of fatty acids and other molecules in peroxisomes
produce H2O2; phagocytosis results in an oxidative burst of nitric oxide (NO ),
which also reacts with superoxide to produce the oxidising and nitrating species
peroxynitrite (ONOO-) (Atta and Choudhary, 2001; Frei, 1994).
"
"
"
"
Free radicals and oxidants are noted to trigger lipid peroxidation, as well as the
oxidation of proteins and DNA, causing extensive damage to body cells.
Oxidative stress resulting from an imbalance of oxidising species and natural
antioxidants in the body has been identified to contribute to aging, cell
crqrvquku."cpf"ugxgtg"fkugcugu"uwej"cu"ecpegt."Rctmkpuqpu"fkugcug."Cn|jgkogtu"
disease, and even cardiovascular disorders (Long et al., 2007; Atawordi, 2005;
Kassab et al., 2003; Favier, 2003). Epidemiological studies and intervention
trials on prevention of cancer and cardiovascular disease in people taking
antioxidant supplements indicate that, dietary intake of antioxidants can help
scavenge free radicals and oxidants and protect the body against diseases (Frei,
1994). It is also proven that most of the dietary antioxidants have low or
minimal toxicity and intake can be increased without adverse effects.
Polyphenols are one of the most copious and widely distributed groups of
secondary metabolites in plants with over 8000 phenolic structures known
(Atawordi et al., 2005; Soobrattee et al., 2005). Naturally, these polyphenols
span from simple molecules of phenolic acids, phenylpropanoids, and
flavonoids to completely polymerised lignins, melanins, and tannins (Kassab et
al., 2003; Giasson et al., 2002). Polyphenols confer a host of biological
"
"
"
"
repercussions
or
effects
such
as
anti-inflammatory,
antiallergic,
Free radical damage is one of the most prominent causes of devastating diseases
that are responsible for killing millions of people in the world, such as
cardiovascular infections, which can manifest as heart attacks and cancer (Amic
et al., 2003; Chaudire and Ferrai-Iliou, 1999). Several research on antioxidants
in biological systems have confirmed the neutralising effects on oxidative stress
that predispose lethal diseases such as atherosclerosis, diabetes mellitus, chronic
inflammation, neurodegenerative disorders and cancerous tumours and thus,
generating keen interest in assessment of antioxidant potentials of consumable
food compounds (Karadag et al., 2009). Free radicals naturally occur in the
body as a result of chemical reactions during normal cellular processes such as
during conversion of food into energy in the body (Oboh and Rocha, 2006).
Antioxidant nutrients in foods soak up all the excess energy of free radicals,
converting them into harmless molecules or waste products that can be
eliminated (Oboh, 2005). Antioxidants are powerful substances that can
neutralise free radicals before they damage the body cells (Oboh and Rocha,
2006). Many phenolics such as flavonoids possess stronger and higher
antioxidant capacities than even ascorbic acid (vitamin C) and tocopherols
(vitamin E). Flavonoids could protect membrane lipids from oxidation. Major
sources of flavonoids are vegetables and fruits (Cai et al., 2004). Current
"
"
"
"
"
"
"
"
"
The DPPH method can be used for solid or liquid samples and is not restricted
or specific to any particular antioxidant component, but applies to the overall
antioxidant capacity of a sample. A measure of total antioxidant capacity helps
understand the functional properties of foods. Antioxidant activity has been
expressed in various ways including percentage of the reagent used, oxidation
inhibition rate and so on. An easier way to present antioxidant activity of foods
would be to reference a common reference standard. One common reference
standard, (S)-(-)-6 hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxylic acid,
also known as Trolox, serves this purpose (Karadag et al., 2009; Brand-Williams
et al., 1995).
"
"
"
"
"
carried out at the Applied Radiation Biology Centre of the Radiological and
Medical Sciences Research Institute (RAMSRI).
"
Accession
Ashanti region
Cape
Eastern region
Asontem-ER
Brong-Ahafo region
Western region
Juaboso
"
"
"
"
"
"
5.3 RESULTS
5.3.1 DPPH radical scavenging assay
Table 5.2 shows concentration of each extract (ethanol and aqueous) against
their percentage inhibition of the DPPH radical scavenger for all 25 accessions
of okra. The coefficient of regression (R2) of the logarithmic trend line was 0.94,
which implies that 94% of the variability in DPPH can be ascribed to the
concentration of the extracts. The analysis of variance (ANOVA) showed that,
concentration significantly (p = 0.05) affected percent inhibition. From the mean
eqorctkuqp."wukpi"vjg" Fwpecpu"ownvkrng"tcpig" vguv"*FOTV+."Citke"ujqtv"htwkv"
(3705.81g/ml), Asontem-GAR (506.49g/ml) gave the highest and lowest
significant DPHH inhibition for ethanol extract compared to the other
concentrations.
Again, the accessions, Kortebortor-ASR (56.33%) and Cape (14.27%) gave the
highest and lowest percent DPHH inhibition for the aqueous extract,
respectively. On solvent extraction basis, ethanol extract recorded a higher total
mean (1417.8613.1319g/ml) of antioxidant activity (AOA) compared to the
aqueous extraction which recorded an AOA value of 1022.1050.22g/ml. This
implies that, ethanol is a better solvent for assessing antioxidant activity in okra
compared to water.
"
"
"
Aqueous Extract
Accession
Asontem-GAR
Asontem-ASR
Asante Type II
%Inhibition
16.74
18.53
32.38
Concentration
506.490.00j
665.020.13ij
1012.001.11fgh
%Inhibition
29.13
27.13
22.06
Concentration
906.690.00efgh
992.432.22efg
678.330.00hijklm
Asontem-ER
21.68
732.71.22hij
23.05
1074.27821.62
Wune Mana
Labadi
Agric Type I
54.060
31.97
27.70
3030.3869.31b
941.631.05fghi
1183.440.76def
48.47
17.94
17.43
2660.700.00b
514.140.00lmn
726.921.52ghijkl
Clemson Spineless
Volta
Agric Short Fruit
45.27
32.14
46.24
1885.32732.38c
1440.871.59de
3705.810.00a
21.65
23.95
25.94
877.830.00efghij
1060.731.59de
2039.890.00c
Debo'
Juaboso
Kortebortor-BAR
51.12
35.01
46.12
3812.160.00a
1206.490.61def
2001.2212.19c
50.84
23.47
20.96
3791.252.61a
796.161.22fghijk
883.920.00efghi
Legon Fingers
Indiana
Asontem-BAR
Mamolega
Nkran Nkuruma
34.89
55.97
44.80
51.15
28.99
1001.851.02fgh
1829.58483.00c
1195.87506.53def
1483.520.00d
763.450.94hij
21.51
24.90
23.71
15.09
18.18
605.542.03klmn
795.550.00fghijk
618.990.00ijklmn
415.5111.18n
467.970.94mn
Atomic
Cape'
Mapelega
Kortebortor-ASR
23.16
25.56
29.81
28.22
942.2213.17fghi
1135.490.00efg
766.310.91hij
643.360.41ij
23.84
14.27
30.43
56.33
971.482.93fg
612.194.77jklmn
782.770.00ghijkl
1309.570.81d
Yeji-Local
Cs-Legon
Asontem N V.
47.21
29.02
22.89
2050.140.15c
805.700.00ghij
705.491.66hij
16.60
25.39
28.01
690.341.52hijklm
700.900.98hijklm
870.660.55efghij
de
sd=standard deviation, mean with same letters in a column are not statistically differepv"*r2027+"
from each other ceeqtfkpi"vq"Fwpecpu"ownvkrng"tcpig"vguvu0
"
"
"
"
fruit) and 122.482.69g/g/QE (Yeji-Local) to 2003.692.55g/g/QE (CsLegon) for the ethanol and aqueous extracts respectively.
The
mean
total
flavonoid
content
(TFC)
for
ethanol
extract
Phenolic contents measured by the Folin Cicalteu reagents in terms of gallic acid
equivalent
(GAE)
ranged
from
25.835.30g/g/GAE
(Debo)
to
The
average
total
phenolic
content
(TPC)
for
ethanol
extract
"
"
"
Phenolics
Asontem-GAR
Concentration
(eth)
2264.980.00gh
Concentration
(aq)
492.899.56k
Concentration
(eth)
11.381.77defghi
Concentration
(aq)
20.940.00fg
Asontem-ASR
2338.878.31gh
493.0814.74k
13.182.30cdef
22.460.64ef
Asante Type II
1339.722.12n
465.122.16l
10.211.84efghi
19.661.39g
hi
efghi
19.760.87g
Asontem-ER
2235.140.89
Wune Mana
4980.501.23b
1449.241.28c
18.192.25b
8.580.84l
Labadi
1958.800.00jk
379.290.96n
9.020.28ghi
19.740.11g
Agric Type I
2420.543.36g
345.521.89o
12.3172.21cdefg
28.211.92d
330.210.94
532.772.11
10.631.20
9.691.17
fghi
27.082.00d
Clemson Spineless
1288.0692.91
Volta
Agric Short Fruit
3204.582.34e
5159.2112.90a
400.222.16m
487.6714.92k
14.592.54cd
24.124.64a
6.820.09l
39.460.15b
Debo'
Juaboso
3387.60499.67d
2086.821.55ij
1402.502.38d
648.163.00g
25.835.30a
11.791.47defgh
29.070.00d
20.161.93fg
Kortebortor-BAR
1515.503.88n
279.7233.92p
15.761.71bc
23.621.33e
efghi
16.920.61hi
19.970.00fg
16.380.00ij
14.251.17jk
16.540.36ij
Legon Fingers
Indiana
Asontem-BAR
Mamolega
Nkran Nkuruma
3601.420.65
1609.011.45mn
1517.550.60n
1181.910.45n
1940.7715.19 jkl
693.073.84
750.8328.82e
485.243.39k
517.912.51j
480.623.88kl
9.890.61
10.510.63efghi
8.551.01hi
12.231.26cdefg
8.890.54ghi
Atomic
988.532.15o
582.501.92i
13.422.22cde
19.560.24gh
Cape'
Mapelega
Kortebortor-ASR
3077.1811.14ef
1874.811.78kl
1774.3322.10lm
179.984.04q
1579.183.84b
1443.880.00c
10.183.27efghi
9.070.53ghi
8.010.37i
12.991.22k
32.275.40c
63.223.95a
Yeji-Local
1595.612.24n
122.482.69r
18.263.03b
13.370.75k
Cs-Legon
Asontem N V.
871.573.84
2993.192.93
2003.692.55
f
608.396.68
10.760.89
efghi
7.970.00l
18.350.79
8.330.00l
sd=standard deviation, mean with same letters in a column are not statistically difhgtgpv" *r2027+"
from each other ceeqtfkpi"vq"Fwpecpu"ownvkrng"tcpig"vguvu0, aq = aqueous, eth = ethanol.
"
"
5.4 DISCUSSION
5.4.1 Total flavonoid and phenolic contents of ethanoic and aqueous
extracts of Accessions of Abelmoschus spp
The mean total flavonoid contents (TFCs) were (2288.2027.75g/g/QE) for
ethanol extract and (686.176.17g/g/QE) for the aqueous extracts. The
antioxidant activity and total flavonoid contents were generally moderate,
affirming that ethanoic extraction method was more efficient. This also indicates
that, flavonoids in pods of okra are more soluble in organic solvents such as
ethanol
than
in
water.
The
highest
total
flavonoid
content
"
flavonoids in ten plants; Chang et al. (2002) who also worked on total flavonoid
in propolis.
Specifically, flavonoids have variously been reported to substantially influence
antioxidant properties in mucilagenous vegetables (Mariani et al., 2008; Firuzi
et al., 2005; Choi et al., 2002; Rice-Evans et al., 1996).
total
phenolic
content
was
registered
by
Kortebortor-ASR
The total phenolic composition of the accessions of okra compares well with
common fruits and vegetables noted for their relatively high phenolic
constituents such as cranberries (52.722.15mg/g), apple (29.630.64mg/g),
strawberries
(16.000.12mg/g),
pineapple
(9.430.15mg/g),
banana
The relatively high radical scavenging impact on the DPPH radicals may be
attributed to high flavonoid and phenolic contents in the extracts of the
accessions (Maltas and Yildiz, 2012; Maltas et al., 2011). There was however, a
discrepancy observed between the ethanol and aqueous extracts, in that, not all
accessions with high TFC(s) or TPC(s) necessarily gave a corresponding higher
scavenging activity, corroborating earlier reports of Owusu-Ansah (2010) for
Moringa, Quartey (2010) for tomatoes and Prior et al. (2005) for dietary
supplements.
5.5 CONCLUSION
The high carcinogenicity nature of synthetic antioxidants has enkindled
considerable interest in finding naturally occurring antioxidants for use in foods,
cosmetics or medicinal materials as possible substitutes for the synthetics (Sasaki
et al., 2002). High antioxidant foods have greater potential to reduce free radicals
in the body than do low antioxidant foods. It is therefore, imperative to ascertain
the antioxidant content of foods such as okra, in addition to knowing the basic
nutritional information such as the vitamin, fibre, fat, mineral and protein
contents (Prakash et al., 2010).
"
II.
There was high variability with respect to TFCs, TPCs and total
activity in okra than in water. Breeders can also explore the possibility of
breeding for high phenolic and antioxidants containing varieties. The strong free
radical scavenging activity of Abelmoschus spp requires further studies for the
isolation and identification of other bioactive compounds inherent in this
species.
5.6 RECOMMENDATIONS
I.
REFERENCES
Abdel-Hameed, E.S.S. (2009). Total phenolic contents and free radical
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"
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"
"
Evid. Based
"
"
CHAPTER SIX
"
6.1 INTRODUCTION
6.1.1 Okra Production in the world
Okra, (Abelmoschus spp L.), is prominently featured in nearly every market in
Africa. In Ghana, it is the fourth most popularly patronised vegetable after
tomatoes, pepper and garden eggs (Oppong-Sekyere, 2011; Lamont Jr., 1999;
Sinnadurai, 1973). The world okra production, as of 2010, was estimated at 6.9
million tonnes with Asia leading the production by 75.92% followed by Africa
(23.41%) and the Americas (0.54%) (Table 6.1) (FAOSTAT, 2012). On the
whole, India is the leading producer of okra with over 70% of the total world
production, followed by Nigeria, Pakistan, Sudan, Ghana, Egypt and Iraq
(FAOSTAT, 2012; Gulsen et al., 2007).
"
Table 6.1: Estimated production of okra in selected geographical regions and countries (metric
tonnes)
Year
World
Africa
Asia
Americas
Ghana
Nigeria
Sudan
India
USA
2010
6917062
1619525
5251415
37122
82500
955600
256000
4803000
8500
2009
6570901
1524558
4985571
51612
71350
826170
252000
4528000
9825
2008
6400222
1722399
4611226
57534
89731
1039000
249000
4179000
9673
2007
6533044
1970326
4495866
58352
108000
1280000
216950
4070000
10000
2006
6191495
1669583
4447523
62839
105000
1000000
168000
3974600
9500
"
"
In Ghana, okra is traded in the fresh state in almost all markets during the rainy
season and in a dehydrated form during the dry season. It is particularly found in
Northern Ghana due to its tremendous commercial potential for resourceconstrained women farmers and its vital importance as a component of various
recipes (Oppong-Sekyere et al., 2012; Torkpo et al., 2006). The pods, which
possess unique flavour and texture, are boiled and added to soups, or sliced and
fried. They may be used alone or mixed with other vegetables in local
preparations.
Mucilage from fruits of okra is known to be a good thickening agent for gravy.
In West Africa, young pods are thinly unkegf" vq" rtgrctg" qmtc" uqwr." c" rgthgev"
partner of hwhw" *Mwoct" et al., 2010; National Academies Press, 2006). Gums
stay intact inside the pods when sliced and dried and remain useful for
flavouring and thickening foods. Okra is sometimes used in place of eggplant in
many recipes in some parts of Asia (Baytop, 1984). The seeds of mature pods
are used for animal feed and for production of edible oil (Oyelade et al., 2003;
Oyenuga, 1969; Busson, 1963).
"
"
"
In West Africa, okra pods are often sliced, sun dried, and ground into powder
for use in the lean season (hungry time) that hits each year just before the new
harvest (Akintoye et al., 2011; Kumar et al., 2010; National Academies Press,
2006). Young immature okra pods can be consumed in different forms
(Ndunguru and Rajabu, 2004), and in variety of cuisines in Asia and Africa. In
the Caribbean islands, okra is cooked up and eaten as soup, often with fish
(www.whfoods.com/genpage.php?pfriendly=1&tname=foodspice&dbid=128).
The pods are generally cut into small circular sections and the immature leaves
are utilised in local cuisines in Nigeria (Ayodele, 1993), and also eaten raw in
salads
in
Papua
New
Guinea,
the
Caribbean
and
Malaysia
(www.whfoods.com/genpage.php?pfriendly=1&tname=foodspice&dbid=128).
In Turkey, the pods are strung out to dry for use in winter and the dried seeds are
used as substitutes for coffee (Calisir et al., 2005; Moekchantuk and Kumar,
2004). The leaf buds and flowers are also edible (Doijode, 2001). Locally, okra
is commonly used in soups and stews. It is also used as a sieving agent in
dtgykpi"qh"rkvq"*cp"kpfkigpous wine) in northern Ghana.
"
Saifullah and Rabbani, 2009; Kahlon et al., 2007). Okra is also a good source of
vitamin B, some essential minerals (Lamont, 1999), rhamnose (22%),
galacturonic acid (27%) and amino acids (11%) (Benchasri, 2012a).
The composition of okra leaf per 100 g edible portion is: 81.50 g water, 56.00
kcal energy, 4.40 g protein, fat 0.60 g, carbohydrate11.30 g, fibre 2.10 g,
ascorbic acid 59.00 mg, -carotene 5:7022" i, thiamine 0.25 mg, riboflavin 2.80
mg, niacin 0.20 mg (Benchasri, 2012a; Varmudy, 2011; Gopalan et al., 2007).
Carbohydrates are mainly present in the form of mucilage (Kumar et al., 2009;
Liu et al., 2005).
"
"
mg/g) (Cook et al., 2000; Glew, 1997). The leaves are also known to contain Ca,
P, and Fe (Benchasri, 2012a&b; Varmudy, 2011; Gopalan et al., 2007).
6.1.5 Significance of mineral elements to human
Manganese helps the body form connective tissues, bones, blood clotting
factors, and sex hormones. It also plays a role in fat and carbohydrate
metabolism, calcium absorption, and blood sugar regulation. Manganese is also
integral to normal brain and nerve function. In addition, it is a component of the
antioxidant enzyme superoxide dismutase (SOD), which helps combat free
radicals
in
the
body
(http://www.umm.edu/altmed/articles/manganese-
of
bones)
in
adults
and
(http://ods.od.nih.gov/factsheets/Calcium-HealthProfessional).
"
"
older
people
and
reducing
the
risk
of
heart
attack
and
stroke
(http://www.whfoods.com/genpage.php?pfriendly=1&tname=foodspice&dbid=1
28). The daily requirement of magnesium for children between the ages of 1-10
spans from 150 to 250 mg, 350 mg for males between the ages of 11 and 14
years. Males between the ages of 15 and 18 require 400 mg while all females
require 300mg. Pregnant women and nursing mothers however, require 450 mg
of
Mg
(http://www.rawfoodexplained.com/minerals/the-minerals-in-the-
body.html).
Copper combines with certain proteins to produce enzymes that act as catalysts
to help a number of body functions. It is also involved in the transformation of
melanin for pigmentation of the skin and helps to form cross-links in collagen
and elastin and thereby maintain and repair connective tissues. This is especially
important for the heart and arteries (Dietary Reference, 1991). Research
suggests that copper deficiency is one factor leading to an increased risk of
developing coronary heart disease. Copper, along with iron, helps in the
formation of red blood cells. It also helps in keeping the blood vessels, nerves,
immune
system,
and
bones
healthy
(http://www.copper.org/consumers/health/papers/cu_health_uk/cu_health_uk.ht
ml). It helps iron-rich foods to form red haemoglobin in the blood. In fact, it is
essential for the normal healthy growth and reproduction of all higher animals.
"
"
"
The daily requirement of copper for both males and females is 0.4-1.2 mg/day
(Dietary Reference, 1991).
Chlorine (Cl) exists primarily as chloride anion that interacts with cations such
as sodium to produce salt (table salt / sodium chloride), and with hydrogen to
produce stomach acid (hydrochloric acid). It is an essential nutrient in human
diet and is necessary for healthy nervous and digestive systems. It is required for
protein digestion (pepsin), Vitamin B12 absorption (intrinsic factor), and
absorption of metallic minerals (http://www.acu-cell.com/fcl.html).
"
"
"
brain
functioning
and
the
production
of
sex
hormones
Aluminium is present in only minute amount in plant and animal tissues and it is
commonly ingested in foods and in medicines. It is found in the lungs, kidneys,
brain, liver, and thyroid. The daily intake of aluminium may range from 10-110
mg, but most of this is eliminated from the body through faeces, urine and some
in sweat. More aluminium is stored, particularly in the bones when there is
decreased kidney function (http://www.healthy.net/scr/article.aspx?Id=1958).
Recommended daily dietary intake of Al is 2.0mg/kg/day (Dietary Reference,
1991).
"
"
"
"
6.1.6 Mucilage
Mucilages are water-soluble polysaccharides inherent in several plant species
and in some microorganisms (National Academies Press, 2006; Woolfe et al.,
1977). Mostly plants belonging to the Malvaceae family and other related
species such as baobab (Adansonia digitata L.), cotton (Gossypium spp),
ambrette (Abelmoschus moschatus Medik.), kenaf (Hibiscus cannabinus),
plantago (Plantago psyllium), aloe (Aloea vera), cocoa (Theobroma cacao),
kapok (Ceiba pentandra), roselle (Hibiscus sabdariffa) and okra (Abelmoschus
spp) are known for their considerable mucilage content (Ahmad et al., 2009;
Cvancara, 2001). Mucilage in okra can however be found in almost all parts of
the crop (Kumar et al., 2010; National Academies Press, 2006). The immature
pods, seeds, leaves, stems, barks, roots, flowers and even the cell walls of the
fruits are noted to harbour considerable amount of mucilage (Sengkhamparn et
al., 2009; Woolfe et al., 1977).
The mucilage and gums in plants are known to aid in water storage, decrease
diffusion in the plant, aid in seed dispersal and germination, and act as a
membrane thickener and food reserve. Plant gums and mucilage, in particular,
have generated tremendous interest due to their varied pharmaceutical
applications, among other uses as diluents, bio-binders, tablet disintegrants,
protective colloids in suspensions, thickeners in oral liquids, gelling agents in
gels, gum substitute, effluents in rheological engineering and bases in
suppository (Okoye et al., 2009; Tomoda et al., 1987; Woolfe et al., 1977).
"
"
"
"
form
and
without
any
pre-treatment
of
the
sample
(www.ne.ncsu.edu/nrp/naa.html#ag).
For certain elements, INAA offers sensitivities that are superior to those possible
by other techniques; approximately parts per billion or better. In addition to the
elemental and isotopic analysis of samples, INAA permits the analysis of nonradioactive tracers introduced into biological, chemical, and/or industrial
processes for process identification and optimisation. It is non-destructive and
therefore, does not suffer from the errors associated with yield determinations; it
has very high sensitivities for most of the elements that can be determined by
INAA - most detection limits span from ~0.05 to ~50 ppm ( 1 ppb for some
high-purity materials). It is highly precise and accurate; permits the analysis of
"
"
samples ranging in volume from 0.1 ml to 20 ml, and in mass from ~0.001 gram
to 10 grams depending on sample density. Samples for INAA can be solids,
liquids, gases, mixtures, and suspensions, hence the choice of this technique for
this study.
ii.
of okra
"
"
"
"
The relative efficiency of the detector is 25% with energy resolution of 1.8 keV
cv"c" -ray energy of 1332keV of
60
vkog."fgvgevqtu"fgcf"vkog"ycu"eqpvtqnngf"vq"dg"nguu"vjcp"32'0" Identification of
-ray of product radionuclide was done through the energies and quantitative
analysis of the concentration was achieved wukpi" vjg" -ray spectrum analysis
software, ORTEC MEASTRO-32.
"
"
"
structures on the harvested pods were cut or peeled off and then washed off any
debris.
Three hundred and fifty grammes (350g) of freshly harvested pods of okra were
thoroughly washed, sliced and chopped. Four hundred and fifty millilitres
(450ml) of distilled water was added and blended for 30 seconds. This was
topped up to 900ml of distilled water and then filtered using a muslin cloth to
extract the mucilage. Three hundred and fifty millilitres (350ml) of distilled
water was added to rinse and also to prevent suspended colloidal solid seepage
and speed up to ease the release of the mucilage from the supernatant.
"
6.3 RESULTS
6.3.1 Concentrations of Nine Essential Elements in Accessions of
Abelmoshus spp L.
Concentrations of five essential macro elements (Na, Mg, Ca, K and Cl), three
micro elements (Al, Mn and Cu) and one trace element (Br) detected by
instrumental neutron activation analysis (INAA) in 22 okra accessions are
shown in Table 6.2. Asontem-BAR registered the highest content (61.529.23
mg/kg) of Mg while Asontem NV had the least (40.096.01 mg/kg) Mg
"
"
"
In addition, the elements Al, Na, Mg, Br, Cu and Cl detected using the technique
of Instrumental Neutron Activation Analysis (INAA), varied considerably
among all accessions under study. Mamolega recorded the highest concentration
(778.511.60 mg/kg) of Na whilst Asontem-ASR registered the least
concentration (16.322.45 mg/kg) of Na. Most of the accessions recorded fairly
high concentration of Na. Mamolega also recorded the highest Al concentration
(77.4112.31 mg/kg) while the least Al concentration was recorded by Indiana
(31.164.67 mg/kg). On the other hand, Indiana registered the highest Mn
concentration (36.355.45 mg/kg) with the least registered by Kortebortor-ASR
(11.002.49 mg/kg). Similarly, Cape gave the highest Cu concentration
(424.563.67 mg/kg), while Kortebortor-ASR once again, recorded the lowest
(85.2612.79 mg/kg).
concentration
of
other
"
elements
analysed.
"
"
Table 6.2: Concentration of Nine Mineral Elements in the Accessions of Okra in mg/kg*
Accession
Asontem-ASR
Macro Elements
Mg
54.758.21
Micro Elements
Ca
Cl
Na
Br
23.803.57
89.2613.39
61.539.23
16.322.45
15.542.33
Al
33.805.41
Mn
15.562.93
Cu
< 0.01
Asontem NV
40.096.01
16.042.41
91.9813.79
81.3912.20
190.128.52
29.964.49
47.967.67
15.682.99
< 0.01
Agric Type I
45.096.76
27.214.08
82.2512.34
54.818.22
189.028.24
20.013.00
32.845.29
14.592.95
< 0.01
Asontem-BAR
61.529.23*
19.812.97
11.131.67
88.1913.23*
191.528.25
31.644.75
37.175.95
14.452.77
186.527.98
Atomic
48.537.28
16.972.55
76.9111.54
36.845.53
362.654.39
51.867.78*
37.345.94
16.473.12
165.624.84
43.966.59
76.4311.46*
73.0910.96
51.377.71
221.4133.21
41.626.24
51.398.17*
17.073.12
< 0.01
Cape
54.708.21
19.482.92
11.711.76
10.851.63
385.657.16*
36.205.43
38.086.13
25.214.84*
424.563.67*
Asante Type II
50.107.52
19.072.86
90.9313.64
71.2810.69
279.340.99
25.653.85
37.766.04
17.073.28
< 0.01
Juaboso
59.598.94
19.622.94
11.691.75
83.4412.52
218.332.75
31.424.72
35.265.64
24.014.29
263.439.51
Labadi
55.768.36
23.143.47
10.631.59
85.5212.83
316.7847.52
27.434.11
41.396.62
20.483.63
276.441.46
46.867.03
21.143.17
94.2314.13*
43.016.45
170.025.50
36.105.42
36.535.88
11.002.49
85.2612.79
Mean
49.997.49
32.064.81
64.319.64
64.739.71
31942.97
34.415.25
42.456.77
17.83.33
63.729.56
CV (%)
23.01
17.33
187.74
269.75
6.14
15.73
60.55
193.13
31.97
RDI
320-420
2000
500-1200
100-500
2400-5175
2-5
2-5
0.4-1.2
Clemson
Spineless
KortebortorASR
"
*Values bolded and asterisked refers to the highest concentration for a particular element; Bolded values represents the lowest concentration for a particular
element. refers to standard error of the mineral element concentration in the accession. CV = Coefficient of Variation; RDI = recommended daily intake:
(Source = Dietary Reference, 1991).
"
"
"
"
"
Macro Elements
Accession
Mg
Micro Elements
Ca
Na
Br
Al
Mn
Cu
46.276.94
521.3278.20
18.452.77
31.164.67
36.355.45*
< 0.01
88.3813.26
72.5410.88
223.433.51
34.755.21
70.1111.15
17.273.51
< 0.01
90.6213.59
65.089.76
570.785.61
34.675.20
43.576.93
15.502.88
< 0.01
52.897.93
336.4250.46
53.187.98
34.695.59
15.892.86
< 0.01
90.913.63
95.4814.32*
778.511.60*
104.615.69*
77.4112.31*
18.033.55
< 0.01
89.4913.42
58.068.71
171.125.67
22.673.40
37.896.06
13.882.58
< 0.01
96.2114.43
93.7814.07
590.488.56
25.723.86
35.965.75
20.353.81
< 0.01
18.312.75
72.5210.87
62.119.32
277.041.55
38.425.76
42.506.80
13.312.64
< 0.01
92. 1413.82
10.341.55
69.1710.37
205.2630.79
21.562.23
42.726.88
15.532.92
< 0.01
13.422.01
68.3910.26
Cl
Indiana
44.576.69
Wune mana
50.967.64
23.333.49
Legon Fingers
48.957.34
15.322.30
Nkran
51.277.69
97.4314.61*
84.7912.71
Mamolega
50.917.64
40.796.12
Kortebortor-
51.477.72
20.093.01
Asontem-GAR
47.597.14
17.882.68
Asontem-ER
42.956.44
Agric
48.577.29
Nkuruma
BAR
Short
Fruit
Yeji-Local
55.718.36
65.479.82
10.121.52
78.8511.83
387.8258.17
46.787.02
50.217.98
22.774.08
< 0.01
Volta
45.946.89
18.452.77
68.6310.29
61.109.17
415.2662.29
28.654.30
38.116.10
11.632.61
< 0.01
Mean
49.997.49
32.064.81
64.319.64
64.739.71
31942.97
34.415.25
42.456.77
17.83.33
63.729.56
CV (%)
23.01
17.33
187.74
269.75
6.14
15.73
60.55
193.13
31.97
RDI
320-420
2000
500-1200
100-500
2400-5175
2-5
2-5
0.4-1.2
"
*Values bolded and asterisked refers to the highest concentration for a particular element; Bolded values represents the lowest concentration for a particular
element. refers to standard error of the mineral element concentration in the accession. CV = Coefficient of Variation; RDI = recommended daily intake:
(Source
"
Dietary
"
Reference,
1991).
"6.3.2:
Table 6.3 shows the association between pairs of elements in the accessions
analysed. Aluminium showed strong association with Bromine and moderate
association with four other elements but was negatively associated with
manganese, copper and magnesium. Magnesium showed strong association with
copper. Manganese exhibited moderate association with copper, sodium and
magnesium but was weakly or negatively correlated with bromine, potassium,
calcium and chlorine. Copper registered a strong positive association with
magnesium but weakly correlated with the rest of the mineral elements detected
among the accessions. Bromine gave a moderate positive association with four
other elements. Strong negative association existed between magnesium and
calcium but the same element exhibited a weak positive association with
potassium and chlorine. There was no correlation between potassium and
chlorine but weak negative correlation with calcium. Calcium recorded a very
weak negative association with chlorine.
Table 6.3: Correlations among Nine Mineral Elements in 22 accessions of Abelmoschus spp*.
Mineral
Element
Al
Mn
Cu
Na
Br
Mg
Ca
Al
Mn
-0.086
Cu
-0.219
Na
0.344
0.371
Br
0.662*
-0.018
0.011
0.589
Mg
-0.042
0.158
0.565
-0.105
0.041
0.173
-0.082
-0.258
-0.053
0.270
0.012
Ca
0.128
-0.342
-0.646
0.115
0.069
-0.615
-0.210
Cl
0.376
-0.098
-0.276
0.178
0.226
0.000
0.291
-0.064
* p = 0.05
"
0.168
-0.057
Cl
"
"
"
"
DKA
366.75a*
Yeji-Local
329.00ab
Amanfrom
316.75abc
Asontem NV.
284.75bcd
Kortebortor-BAR
269.75bcd
Atomic
247.75cde
Akrave
246.25cde
Asontem-ER
221.75def
Nkran Nkuruma
182.00efg
Asontem-ASR
165.50fgh
Juaboso
162.25fghi
Legon fingers
160.75fghi
Mapelega
148.00ghij
Volta
133.00ghijk
116.00ghijkl
Cs-Legon
103.75hijkl
Labadi
97.25hijkl
Asontem-GAR
91.75ijkl
Asante type II
88.25jkl
Asontem-BAR
64.25kl
Ecrg
53.00l*
Mean
183.26101.19
CV (%)
55.22
*Accession with highest mean maximum viscosity is bolded and underlined and
accession with lowest mean maximum viscosity is bolded. = standard deviation of
mean; bu = Brabender unit.
"
6.4 DISCUSSION
6.4.1 ASSESSMENT OF ESSENTIAL MINERAL ELEMENTS IN OKRA
ACCESSIONS
Determination of the mineral element composition of the okra accessions brings
to light the extent of genetic variations among the accessions. There was high
variability among the okra accessions for concentrations of all five essential
macro elements in the edible fruits as detected using INAA. Concentration of
sodium recorded ranged from 778.511.60 mg/kg to 16.322.45 mg/kg for
Mamolega and Asontem-ASR respectively. The Report by Kumar et al. (2010)
that okra pods contain appreciable levels of Na is consistent with the result of
this study. The high concentration of Na as revealed in this study could also be
explained by the nutrient status and anthropogenic activities on the soils at the
experimental site.
Potassium concentration on the other hand was equally appreciable for most of
the accessions. Concentration values recorded in this study were above what was
reported by Benchasri (2012a) in okra and Adotey et al. (2009) in tomato.
Indiana recorded the lowest of 13.422.01 mg/kg; while Nkran Nkuruma
registered the highest concentration of 97.4314.61 mg/kg. The recommended
daily
intake
for
potassium
is
200mg/kg
"
"
"
Concentrations of all the micro elements (Al, Cu, Mn and Br) detected were
considerably high. Quartey (2010) and Adotey et al. (2009) reported seemingly
high values for these elements in tomato, concurring concentrations of these
elements in this study. Al contents recorded for all accessions were
"
"
"
"
comparatively higher than clover (23.6 mg/kg), oats (7.3 mg/kg), bean (5.1
mg/kg), green lentil (6.6 mg/kg), spinach (12.6 mg/kg), corn (11.90 mg/kg), red
lentil (3.7 mg/kg) and rice (8.4 mg/kg) as reported by Hicsonmez et al. (2012).
The significant variable concentrations of Al in all accessions may partly be
explained by the genetics of the individual accessions and the influence of the
soil at the experimental field. Children, women of reproductive age and pregnant
women are most vulnerable to micro nutrient deficiency and anaemia (Ghana
Demographic Health Survey, 2004); hence, diets for this group of persons ought
to be rich in micro nutrients. Okra could serve as source of Al, Mn and other
essential micro nutrients in the diets of children and pregnant women in Ghana.
Copper was however not detected for most accessions, this could be ascribed to
low prevalence level of Cu in these accessions.
"
"
"
"
"
"
"
Legon fingers may be selected for future hybridisation work aimed at further
improvement in mineral element constituents alongside fresh fruit yield and
other desirable traits.
Research indicates that cultivars belonging to the West African Taxon (WAT)
mostly A. caillei are high in mucilage content (National Academies Press, 2006).
DKA (366.8 bu), Yeji-Local (329 bu), Amanfrom (316.8 bu), Asontem NV
(284.8 bu) and Kortebortor-BAR (269.8 bu) met all the requirements of
Abelmoschus caillei. Stev. (WAT) in terms of their morphological
characteristics. These accessions were hardy and perennial in nature with stout
"
"
stems and flattened broad leaves (Table 3.3). They flowered very late and
produced slimy pods, which were non-uniform in shape and size (Table 3.3),
compared to the other accessions. These accessions could be employed as source
of genes for breeding okra with higher mucilage content (Kumar et al., 2010).
These accessions with high level of mucilage were generally high yielding, late
maturing with erect growth habit (Table 3.2; chapter 3). Accessions from the
Volta, Ashanti and Brong Ahafo regions recorded equally high viscosity with
moderate level of mucilage. These may be attributed to the feeding habits of
inhabitants of these regions and their preference for slimy varieties. The people
of the Volta region and certain tribes (Ewe(s), Krobo(s), the Brong(s),
Dagarti(s), Frafra(s)) in Ghana are known to patronise okra varieties with
variable level of sliminess with correspondent high yielding potential. Hence,
the accessions may have been deliberately selected for by farmers of these
regions to meet preference (s) of consumers. The Bulk of okra from Ghana is
produced from the Brong Ahafo, Volta, Northern, Greater Accra and Central
regions (Torkpo et al., 2006; NARP, 1993); however, farmers cultivate different
varieties to meet local market demands in these regions. Some local consumers
prefer slimier varieties for their preparations while others prefer less slimy
varieties. In addition, growth habits of these varieties (accessions) could have
kphqtogf" hctogtu" ugngevkqp" kp" vjgug" tgikqpu" cu" oquv" qh" vjgug" ceeguukqpu" ykvj"
high viscosity (mucilage) values exhibited erectile growth habit, hardy, stocky
stem and medium to tall plant height. These accessions (DKA, Yeji-Local,
Amanfrom, Asontem NV, and Kortebortor-BAR) were less influenced by the dry
period and even flowered when the rainfall pattern became erratic (November-
"
"
"
2011 to January-2012) and may be selected for further improvement for high
sliminess (viscosity) and for cultivation in areas with erratic rainfall patterns.
On the other hand, accessions with low sliminess (mucilage) were generally
early maturing, short in height, exhibited branched growth habits with short
harvesting duration (single harvest). Greater Accra and other coastal regions in
Ghana have a fluctuating rainfall pattern, which probably dictated hctogtu"
choice in these regions for the type of okra varieties cultivated (accessions).
Cape, Asontem-BAR, Asante type II, Asontem-GAR, Labadi, and Juaboso may
be selected for further improvement as varieties with low mucilage (sliminess)
content.
6.5 CONCLUSIONS
a)
Nine essential mineral elements were detected among the accessions with
varying prevalence levels indicating high level of genetic variability among the
accessions with respect to concentrations of these elements in their edible fruits.
b)
above their recommended daily dietary intake (RDI); therefore, excess intake of
these varieties could pose health risk to consumers. However, concentrations of
major elements such as Na, Mg, K, Ca and Cl were below their RDI (s), hence
intake of these varieties ought to be increased in order to meet RDI(s) for these
essential mineral elements by consumers.
c)
potential, uniform pod shape and size with minimum level of sliminess also
"
"
recorded correspondent optimum concentrations of Al, K, Mn, Cu, Na, Mg, and
Ca.
d)
may be selected as parents for future breeding work with increased and/or
optimum level of essential elements (K, Na, Ca, Mg, Cl), high mucilage
(sliminess), with good yield potential.
e)
The means of maximum viscosity ranged from 53.0 - 366.8 bu for all
accessions studied.
g)
high amounts of mucilage compared to those belonging to the Asian taxon (A.
esculentus).
"
"
"
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"
"
"
"
"
CHAPTER SEVEN
"
7.1 CONCLUSIONS
Immense genetic variations have been observed in okra (Abelmoschus spp L.)
particularly the West African type, Abelmoschus caillei but ironically no serious
breeding effort have yet been committed to exploit and harness the proven
genetic richness to advance the improvement of the crop. Previous international
efforts have been limited to intensive cultivation with increased production base
in a short period (early maturing) and resistance to insects and diseases. There is
still enormous scope for cultivar improvement in Africa for the commercial
sector where hardy, robust, high to moderate mucilage and nutrient content,
long-lived types are required. Germplasm characterisation of 29 indigenous
cultivars (accessions) was done to augment efforts towards improvement of the
crop in Ghana. The following conclusions can be drawn from this study:
1.
"
"
"
parameters such as 50% flowering, 50% fruiting, average number of pods per
plant, first-flowering node, and first fruiting node, maximum plant height and
maximum number of internodes.
(e) Different characters contributed differently to overall genetic variability
among the accessions (PCA) under study.
2
(a) The ISSR primers yielded low level of polymorphism among the 29
(a) There was high variability in the amounts and levels of total
flavonoids, total phenolics and total antioxidant activity, making okra a good
source of natural antioxidants.
(b) Ethanol extraction solvent yielded better antioxidant activity than aqueous
(water) solvent.
(c) Mucilage levels were highly variable in all accessions. DKA, Amanfrom,
Asontem NV, Yeji-Local and Kortebortor-BAR recorded high mucilage content.
(d) Nine essential mineral elements were detected among the accessions with
varying concentrations among the accessions.
(e) There were strong positive associations existing between most pairs of
elements contained in the accessions of okra.
"
"
"
"
7.2 RECOMMENDATIONS
For a co-dominant marker to be employed in any characterisation work, the
sequence of the genome of the plant must be deciphered and known. To the
best of my knowledge, the genome of okra has not yet been sequenced
hence no SSR primers are available now to conduct a comprehensive and
thorough genetic diversity study at the gene level. Sequencing calls for
huge financial and infrastructural investments, therefore I recommend the
following;
(a)
of Indiana, Yeji-Local, Asante type II and Atomic into landraces that are well
adapted to the local agro ecological conditions but lack these qualities (narrow
shape and size, smooth pod pubescence, yellowish green in colour, early
fruiting, short peduncle, moderate level of mucilage, persistent epicalyx
segments, medium pod length and less bulky in weight).
(c)
Local, Asante type II, Cs-Legon, Mamolega, and Atomic were the topmost
accessions with desirable qualities and could be utilised for developing
commercial variety for the export market in Ghana.
(d)
should be extended to other regions and base collections in the genebank of the
"
"
"
"
nutritional compositions should be assessed in both the leaves and the pods.
"
"
"
"
8.0 APPENDICES
"
Accession Identifiers
Collection Descriptors
Source of collection
"
"
"
"
d.f.
BLOCK
s.s.
m.s.
0.17365
0.04341
v.r.
F pr.
0.70
0.595
ACCESSION
28
449.11176
16.03971 258.29
Residual
83
5.15425
0.06210
Total
115
454.43966
3.95165
< 0.001
d.f.
s.s.
m.s.
BLOCK
19.78448
4.94612
ACCESSION
28
Residual
83
3.51632
0.04237
Total
115
33475.19828
291.08868
33451.89748
1194.71062
v.r.
116.75
F pr.
< 0.001
28200.23
< 0.001
v.r.
F pr.
d.f.
BLOCK stratum
s.s.
m.s.
0.15411
0.05137
0.88
335.45909
5742.80
ACCESSION
28
9392.85464
Residual
84
4.90676
"
0.05841
"
<.001
"
"
Total
115
9397.91552
d.f.
s.s.
m.s.
v.r.
BLOCK stratum
F pr.
7.379
2.460
1.94
ACCESSION
28
23832.690
851.167
670.58 <.001
Residual
84
106.621
1.269
Total
115
23946.690
d.f.
s.s.
m.s.
v.r.
BLOCK stratum
1.4483
0.4828
0.80
TREATMENT
28
37505.4483
1339.4803
2225.77
Residual
84
50.5517
0.6018
Total
115
37557.4483
F pr.
<.001
d.f
s.s.
m.s.
v.r.
BLOCK stratum
97.50
32.50
0.35
TREATMENT
28
87351.43
3119.69
Residual
84
7884.71
93.87
Total
115
95333.64
"
"
"
33.24
F pr.
<.001
"
"
d.f.
s.s.
m.s.
BLOCK stratum
0.16524
0.05508
ACCESSION
28
567.00834
20.25030
Residual
84
2.62686
0.03127
Total
115
569.80044
v.r.
F pr.
1.76
647.55 <.001
d.f.
BLOCK stratum
s.s.
m.s.
0.09483
v.r.
0.03161
1.23
2881.03
ACCESSION
28
2069.70690
73.91810
Residual
84
2.15517
0.02566
Total
115
2071.95690
F pr.
<.001
"
Appendix 8.2.9: Variate: TOTAL NUMBER OF LEAVES PER PLANT
(NLPP)
Source of variation
d.f.
s.s.
BLOCK stratum
7.931
m.s.
2.644
2.13
107.89
ACCESSION
28
3742.759
133.670
Residual
84
104.069
1.239
115
3854.759
Total
"
"
v.r
F pr.
<.001
"
"
d.f.
s.s.
m.s.
BLOCK stratum
2.716
0.905
0.64
336.51
TREATMENT
28
13239.966
472.856
Residual
84
118.034
1.405
Total
115
v.r.
F pr.
<.001
13360.716
d.f.
BLOCK stratum
TREATMENT
s.s.
m.s.
v.r.
F pr.
0.50828
0.16943
4.17
28
296.86828
10.60244
261.04
Residual
84
3.41172
Total
115
300.78828
<.001
0.04062
d.f.
s.s.
BLOCK stratum
15.931
5.310
TREATMENT
28
3070.052
109.645
Residual
84
598.569
7.126
115
3684.552
Total
"
m.s.
"
v.r.
F pr.
0.75
15.39
<.001
"
"
d.f.
s.s.
BLOCK stratum
24.276
8.092
4.13
28
380.310
13.583
6.93
164.724
1.961
TREATMENT
Residual
84
Total
115
"
569.310
"
m.s.
v.r
F pr
<.001
"
"
Sequence
C4
GAGAGAGAGAGAGAGYT
O4
GAGAGAGAGAGAGAGAYA
O9
ATGATGATGATGATG
O11
AGAGAGAGAGAGG
O12
GTGGTGGTGGTGGTG
UBC888
BDBCACACACACACACA
"
Accession
Code
Accession
1=ATM
Atomic
16=KBR
Kortebortor-BAR
2=AKV
Akrave
17=IDA
Indiana
3=YJL
Yeji-local
18=DBO
Debo
4=AT2
Asante type II
19=ASF
5=CSP
Clemson spineless
20=AAR
Asontem-ASR
6=NKN
Nkran nkuruma
21=LBI
Labadi
7=CSL
Cs-Legon
22=MLA
Mapelega
8=ABR
Asontem-BAR
23=WMA
Wune mana
9=KPV
Kpeve
24=MLA
Mamolega
10=JBO
Juaboso
25=CPE
Cape
11=LGF
Legon fingers
26=KAR
Kortebortor-ASR
12=AGR
Asontem-GAR
27=VTA
Volta
13=ANV
Asontem nv.
28=AER
Asontem-ER
14=AT1
Agric type I
29=DKA
15=AFM
Amanfrom
DKA
Figure 4.1(a&b).
"
"
"
"
d.f.
s.s.
m.s.
v.r.
Replication stratum
1799.
600.
0.22
Accession
20
687709.
34385.
12.86
Residual
60
160430.
2674.
Total
83
849938.
"
"
F pr.
<.001
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