International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 4, Issue 6, De c 2014, 61-80
TJPRC Pvt. Ltd.

EFFECT OF PUTRESCINE AND SALICYLIC ACID ON AJWAIN PLANT

(TRACHYSPERMUM AMMI) AT VEGETATIVE STAGE GROWN UNDER DROUGHT
STRESS
ZEID, F. A.1 , OMER, E. A.2 , AMIN, A. Y.3 & HANAFY, SHAIMAA A. H. 4
1, 3, 4

Plant Physiology Section, Depart ment of Agricultural Botany, Faculty of Agricu lture,
Cairo University, Giza, Egypt

Department of Medicinal and Aromat ic Plants Research, National Research Centre,

Dokki, Ad Doqi, Giza, Egypt

ABSTRACT
This study investigated the effect of putrescine and salicylic acid on growth, physiology and secondary metabolite
production in Desi Ajwain plant, at the vegetative stage, grown under drought stress. The experiment was designed as a
split plots with 27 treatments and three replications. The main treatments were three irrigation treatments (7, 5, 3 times,
irrigated every 20, 30, 40 days, respectively) with nine sub-treatments; control- spray with water, putrescine (1 ppm)
applied once (P1) and twice (P2), as well as salicylic acid (100 pp m, S1), (150 pp m, S2) and their combinations (P1S1,
P1S2, P2S1, P2S2), which were applied at 45 and 60 days fro m planting (DFP). Results indicated that drought stress
significantly reduced all roots and shoot morphological growth characters, as well as shoots: root ratio. While total sugars,
total soluble phenols, and free amino acids in shoots and roots were significantly increased. In addition, chlorophyll b was
significantly decreased. However, no significant differences were found among irrigation treatments on chlorophyll a, total
chlorophylls and total carotenoids. In addition, increasing irrigated intervals to 40 days increased the essential oil %,
whereas, oil content per plant increased by irrigation every 30 days. Meanwhile, it was found that P1 and S2 application
enhanced the all shoot growth characters and root dry weight than P2 and S1, respectively. On the other hand, the
application of S1 was enhanced the root length and root fresh weight than S2. The total soluble sugars were increased by
P2 and P2S1 in plants irrigated every 40 days. In addition, P1, S2 and P1S2 recorded the highest total soluble phenols in
severe drought stressed plants. Furthermore, P1, P2 as well as P2S1 significantly increased free amino acids in severe
drought stressed plants. In addition, chlorophyll a and total chlorophyll were significantly increased with putrescine twice
application (P2). On the other hand, this treatment decreased the chlorophyll b and total carotenoids, whereas the
chlorophyll b and total carotenoids were significantly increased with P1S2 and P2S2 treat ments. The oil % and oil content
per plant as well as oil yield per Feddan significantly increased by S1 and S2 in those irrigated every 40 days. terpiene
was found to be the main constituent and followed by pcymeme. They were increased under water stress, whereas P2
increased thymol by 109% than control. In addition, P2S1 and P2S2 recorded the highest oxygenated and non-oxygenated
compounds in plants irrigated every 40 days. Furthermore, the herb fresh and dry yields were recorded the highest v alues
in the plants irrigated every 30 and 40 days and treated by S2 and S1, respectively.

KEYWORDS: Ajwain Plant (Trachyspermum ammi), Drought Stress, Putrescine, Salicylic Acid, Morphological
Characters, Pig ments Co mposition, Yield, Essential Oil, GC-MS

INTRODUCTION
Desi Ajwain (Trachyspermu m ammi L.) is an aro matic herb. It belongs to family Apiaceae (Umbelliferae).
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Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

This plant has several med ical uses. It is very widely grown in black soil, particularly along the river banks in Egypt as
well as many other countries like India, Iran and Afghanistan as mentioned by Boskabady and Shaikhi (2000).
It is highly esteemed as a remedial agent for flatulence, flatulent colic, tonic dyspepsia, diarrhoea - in short, as a digestive
aid and also as an antiseptic (Cragg and Newman (2005). The Essential oils of Umbelliferae family are present under the
form of tiny vesicles, wh ich located between the cells, where they act as hormones, regulators and catalysts in the
vegetable metabolis m. They seem to help the plant to adapt to its environment and are consequently produced in higher
quantities when plants meet ext reme conditions, biotic and abiotic stresses such as drought danger.
The water status in Egypt has a critical situation. It is facing an annual water deficit of around 7 b illion m 3 .
In fact, United Nat ions is already warning that Egypt could run out of water by the year 2025. The increase in population
and water supply shortages because the limited resource becomes a critical constraint on water future use in Egypt.
In addition, Ethiopia Dam shapes a great future problem for Egypt water availability fro m Nile River. Con sequence, the
agriculture and other sectors development are in threat which endanger food supplies and would lead to aggravate rural
poverty. The government policy is to increase the agricultural land area and water use efficiency. According to the FAO
(2002), the great challenge for the coming decades will therefore be the task of increasing food population with less water,
especially in arid and semi-arid regions.
Drought stress alters the plant photosynthesis, respiration, translocation, ion uptake, carb ohydrates, nutrient
metabolism and hormones as mentioned by Lisar et al. (2012). Drought stress is also known to increase the secondary
metabolite production in a variety of medicinal p lants, like artemisin in in leaves of Artemisia annua as mentioned by
Charles et al. (1993) and ajmalicin in Catharanthus roseus roots as suggested by Jaleel et al. (2008). Also Azhar et al.
(2011) found that cultivation of a medicinal p lant like Desi Ajwain (Trachyspermum ammi) in water-deficient areas would
increase its defense system and the level of act ive co mpounds.
The diamine putrescine (PUT) and its derivatives, the triamine spermidine (SPD) and the tetramine spermine
(SPM ) are the most common polyamines and they have been reported to be imp licated in a variety of plant metabolic and
physiological functions (Kakkar et al. (2000)). Changes in PA, concentrations is a common plant response to a variety of
abiotic stresses, including salinity, high or low temperatures, and drought, as well as biotic stresses (Bouchereau et al.
(1999)). Moreover, Galston and Sawhney (1990) added that the polyamines (PAs), putrescine (Put), spermidine (Spd) and
spermine (Sp m) have been shown to be involved in a variety of plant growth and developmental processes, including cell
division, vascular differentiat ion, root in itiation, shoot formation, flo wer init iation and development, fruit ripening and
senescence and embryo format ion in tissue cultures.
Salicylic acid had been found in many plants (Abdollahi et al. (2011)) and is considered as a quasi-hormonal
substance that plays a crucial ro le in the growth and development of p lants. The axial ro le of salicylic acid in t ranslating
messages as a defence response against pathogenic factors is well known. Salicylic acid also plays an active role in
controlling transpiration, stomata closure, seed germination, fru it yield, g lyc olysis, flowering, heat generation and heat
tolerance.
This study aimed to decrease the number of irrigated water required by the plant and to investigate the effect of
some water stress treatments on growth, yield, and active constituents of Ajwain plant and improving the plant tolerant to
some extent to overcome the hazard effect induced by drought stress using putrescine and/or salicylic acid.

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

63

MATERIALS AND METHODS

Two field experiments were carried out during the two successive seasons 2011/2012 and 2012/ 2013 at the
Agricultural Research and Experimental Station, Faculty of Agriculture, Cairo Un iversity, Giza, Egypt. The study
investigated the effect of foliar applications of salicylic acid and putrescine on Ajwain plant subjected to a drought stress.
The Desi Ajwain (Trachspermu m ammi) seeds, used in this study, were obtained from Suraj, Loblaws INC, Canada. Seeds
were sown at 15 Oct in the two successive seasons. The study included 27 treatments, which was designed as a split plot
experiment, with three replications. The treatments contained three irrigation treatments (seven irrigated times, every 20
days, five irrigated times, every 30 days and three irrigated times, every 40 days) which were considered as the main plots.
The subplots were 9 treatments represented application of putrescine (0, once and twice) at 1 ppm, and/or salicylic acid
(0, 100 ppm and 150 ppm) at 45 and 60 (Days fro m p lanting) DFP. Each plot consisted of three rows (3 m length and 60
cm width). All the agronomic reco mmended practices for Apiaceae family were applied. The plants were fertilized by 120
kg calciu m super phosphate / Fadden (15.5% P 2 O5 ) and 250 kg/fed ammon iu m sulphate (20.5 %N) and 120 kg/fed of
potassium sulphate 48%. A ll fertilizer was added at 30, 45 and 60 days from sowing. So il samp les were randomly taken
each year before cult ivation at the depth of 0-30 cm and were subjected for physical and chemical analysis according to
Jackson (1967). The mean values of mechanical and chemical characters of the experimental soil fo r both seasons of the
soil are shown in Tab le 1.
Table 1: Mechanical and Chemical Analysis of the Soil Experi mental Site
Mechanical Analysis
Coarse
Sand %
1.5 6.0

Fine Sand %

Silt %

Clay %

Texture Cl ass

37.0 41.2

22.0 26.1
31.2 35.0
Clay loam
Chemical anal ysis
pH EC
Soluble Anions (meq/l)
Soluble Cati ons (meq/l)
HCO3 Cl-1
SO4 Na+1
K+1
Ca+2
Mg +2
7.9
2
0.6
2
3.7
3.8
0.2
2
0.3
Mechanical Analysis
Fiel d Capacity
Soil Depth (cm)
Wilting Point %
Available Water %
%
0-20
38.78
16.19
22.59
20-40
38.43
17.36
21.07
40-60
36.34
17.22
19.12
In both successive seasons, samples represented each treatment in the vegetative growth stage, were collected
randomly at 1st January (75 DFP). Three plants were sampled, which represented one replicate, for growth parameters and
chemical analysis. For the morphological characters, shoot height (cm), shoot fresh and dry weights (g), and root fresh and
dry weights (g) as well as the number of branches /plant. The shoots and roots were dried a
was obtained, and then the dry weight was recorded. In addition the chemical analysis, it included the following;
Determinati on of the Essenti al Oil (ml/100 g) of each samp le was determined with hydro-distillation for 3 hours
at Clevenger-type apparatus using fresh herb according to the Egyptian Pharmacopoeia (1984). The resulted essential oil
of each treat ment was separately dehydrated with anhydrous sodium sulphate and kept in the deep freezer until GC -MS.
Essential oil content (ml / plant) was calculated for each treatment by mu ltiplication the fresh weight of plant by essential
oil percent. Essential oil y ield (L/ fed.) was calculated by mult iplication the yield of fresh herb (Ton / fed.) for each
treatment by the essential oil percentage for each treat ment. The GC-Ms analysis of the essential oil samples was carried
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64

Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

out in the first season using gas chromatography-mass spectrometry instrument stands at the Laboratory of Medicinal and
Aromatic Plants, National Research Centre with the following specifications. Instrument: a TRACE GC Ultra Gas
Chro matographs (THERM O Scientific Corp, USA), coupled with a THERMO mass spectrometer detector

(ISQ

Single Quadrupole Mass Spectrometer). The GCM S system was equipped with a TG-WAX MS colu mn (30 m x 0.25 mm
i.d, .25 m f m

). A

rr d

rr r

f 1. mL/ m

ratio of 1:10 and the following temperature program: 40C for 1 min; rising at 4.0 C/ min to 160C and held for 6 min;
rising at 6C/ min to 210C and held for 1 min. The injector and detector were held at 200 and 200C, respectively.
D

mp

(1:1

, v/v) f .2 L f

mx r

d. M

ned by electron

ionization (EI) at 70 eV, using a spectral range of m/z 40 -450. Most of the compounds were identified using two different
analytical methods: (a) KI, Kovats Indices in reference to n -alkanes (C9-C22) (National Institute of Standards and
Technology 2009); and (b) mass spectra (authentic chemicals, Wiley spectral library collection and NSIT library)
confirmat ion of the identified components of the essential oil was carried out by matching with the National Institute of
Standards and Technology (NIST) and the published data by Adams, 2007.

Preparation of Ethanol extract of shoots was used for the determination of total sugars by using the
phenol-sulphuric method according to Dubois et al. (1956), total soluble phenols were estimated using the
folin-Ciocalteau colorimetric method (Swain and Hillis (1959)) and total free amino acids were determined by
using Ninhydrin reagent according to Moore and Stein (1954).

Plant Pigments, the photosynthetic pigments (chlorophyll a, chlorophyll b, total chlo rophylls and total
carotenoids) were determined in leaves samples (mg/g fresh weight) by using dimethyl-formami de according to
Nornai (1982).

Yiel d: The yields of fresh herb (ton/fed), dry herb (ton/fed), seeds (Kg/fed) and essential oil (Lit/fed) were
calculated.
Statistics Analysis, Data collected was subjected to the proper statistical analysis of variance of split plot design

according to the procedures outlined by Snedecor and Cochran (1980). LSD at 5% level of significance was used to
compare treat ments means. All statistical analysis was performed by using analysis of variance technique of
(Mstat-C, 1989) Co mputer software package.

RESULTS AND DISCUSSIONS

Growth Characters
The mean values of growth characters of Ajwain plants as affected by irrigation treatments (every 20 days, 30
days and 40 days), putrescine and salicylic acid are shown in Table 2. All studied treatments showed significant effect on
all gro wth characters of Ajwain plants except the number of branches. I generally, increasing the irrigation intervals
(decreasing the number of irrigation times) decreased all gro wth parameters. Irrigation every 40 days decreased all the
roots and shoots -growth characters, as well as shoots to root ratio. While those irrigated every 30 days recorded the
highest values of the shoot height and shoot fresh weight, however, the highest significant value in root growth characters
as the root length, fresh and dry weights of roots, as well as shoot dry weight, and shoot: root ratio were re corded by the
plants, which irrigated every 20 days . Neseim et al. (2014) worked on sugar beet ( Beta vulgaris) added that Plant fresh
weight was reduced under drought stress which could be attributed to the foliage and root fresh weight reduction, which

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

65

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

was related to the turgor pressure fall, leading to a decrease in cellular expansion and resulted in a reduction in leaf area
and size. Consequently, the exposed leaf area to the light is decreased leading to a reduction in dry matter light conversion
coefficient. Azhar et al. (2011) found that herb fresh and dry weights were reduced by increasing drought stress levels. No
significant differences in number of branches were found among the studied treatments.

The evidence of the

plant cope the drought stress is supported by the difference between plants irrigated every 20 and 30 days were less than 50
% as Staniszewska et al. (2003) confirmed, and they mentioned also, the reduction in herb due to the low mo isture
availability around roots, restricted proliferation of root biomass or limited absorption of nutrients.

In addition,

the shoot to root ratio was decreased in plants irrigated every 30 and 40 days, as the shoot fresh and dry weights reduced
more than root fresh and dry weights. Manivannan et al. (2007) mentioned that the increase in shoot to root ratio under
drought conditions was related to ABA content of roots and shoots.
Table 2: Shoot height, Root leng th, Fresh and Dry Weights of Shoot and Root and Shoot: Root Ratio as well as
Number of Branches of Ajwai n pl ant as Affected by Irrigation, Putrescine, Salicylic aci d and their Interacti ons
during Vegetati ve Stage (Combined of Seasons 2011-2012 and 2012-2013)
Shoot fresh weight (g)
Mean
Irrigation Times
(B)
7
5
3

Character
Treatment
Control
P1
P2
S1
S2
P1S1
P1S2
P2S1
P2S2
Mean A
LSD
Character
Control
P1
P2
S1
S2
P1S1
P1S2
P2S1
P2S2
Mean A
LSD

Shoot dry weight (g)

Mean
Irrigation Times
(B)
7
5
3

Root fresh weight (g)

Irrigation Times
Mean (B)
7
5
3

Root dry weight (g)

Mean
Irrigation Times
(B)
7
5
3

163.71

49.67

25.17

79.52

19.65

6.38

3.52

9.85

4.83

3.70

3.56

4.03

0.58

0.48

0.50

0.52

88.61

103.47

21.81

71.30

10.90

13.76

3.12

9.26

4.89

4.10

3.45

4.15

0.60

0.55

0.49

0.55

74.78

34.86

20.93

43.52

9.42

4.74

3.05

5.74

4.55

3.47

3.58

3.87

0.57

0.47

0.52

0.52

134.96

59.33

26.74

73.68

16.20

7.71

3.74

9.22

5.10

4.02

3.65

4.26

0.61

0.52

0.51

0.55

139.53

50.07

21.99

70.53

17.16

6.66

3.14

8.99

5.19

3.85

3.47

4.17

0.64

0.51

0.50

0.55

67.59

44.70

18.39

43.56

8.31

5.94

2.63

5.63

4.80

3.95

3.40

4.05

0.59

0.53

0.49

0.53

94.21

61.28

21.19

58.89

11.49

8.09

3.01

7.53

4.79

3.83

2.72

3.78

0.59

0.51

0.39

0.49

69.07

30.74

21.96

40.59

8.70

4.18

3.21

5.36

3.66

3.52

2.91

3.36

0.46

0.48

0.42

0.45

82.33

44.53

23.61

50.16

10.13

5.92

3.38

6.48

3.68

4.32

2.92

3.64

0.45

0.58

0.42

0.48

101.64

53.18

22.42

12.44

7.04

3.20

4.61

3.86

3.30

0.57

0.51

0.47

B=13.61

AB=23.57

A=1.00

B=1.74

AB=3.01

A=0.22

B=0.39

AB=0.66

A=0.03

B=0.05

AB=0.09

A= 7.86

Shoot Height (cm)

Root length (cm)

Shoot root ratio

Number of branches

45.82

42.99

39.67

42.82

16.11

12.33

11.88

13.44

34.30

14.40

6.88

18.53

29.01

21.09

18.76

22.95

40.24

44.86

37.89

41.00

16.30

13.67

11.50

13.82

18.13

25.36

6.52

16.67

24.31

26.00

21.98

24.10

34.89

36.33

35.89

35.70

15.18

11.56

11.94

12.89

16.53

9.79

5.95

10.76

16.30

27.42

20.25

21.32

39.67

43.39

39.94

41.00

17.01

13.39

12.16

14.19

26.41

15.44

7.48

16.44

31.83

24.30

18.72

24.95

40.70

43.89

41.50

42.03

17.30

12.83

11.58

13.90

26.79

13.28

6.40

15.49

27.97

24.61

13.98

22.19

38.11

40.64

39.83

39.53

16.00

13.17

11.34

13.50

14.16

12.00

5.55

10.57

26.76

28.37

19.47

24.87

42.11

44.56

41.11

42.59

15.98

12.78

9.06

12.60

19.60

16.36

8.08

14.68

20.08

35.03

17.30

24.14

35.38

36.89

32.39

34.89

12.20

11.72

9.71

11.21

19.15

8.91

8.28

12.11

12.90

19.70

13.09

15.23

31.94

42.67

37.43

37.34

12.26

14.39

9.72

12.12

22.43

10.60

8.26

13.76

13.36

21.93

15.93

17.07

38.76

41.80

38.43

15.37

12.87

10.99

21.94

14.02

7.04

22.50

25.38

17.72

A=1.57

B=2.72

AB=4.71

A=0.74

B=1.28

AB=2.21

A= 1.93

B= 3.33

AB= 5.77

A= ns

B= ns

AB= ns

The effect of either putrescine or salicylic acid revealed that application of putrescine once showed the highest
shoot dry weight. Its application two times decreased all root and shoot growth characters. Likewise, El-Tohamy et al.
(2008)) reported that putrescine spraying increased all growth parameters. Application of salicylic acid at concentration of
150 pp m (S2) significantly increased the shoot growth characters, the fresh and dry we ights of the shoots, shoot height as
well as root dry weight, which were in harmony with El-Tarawy et al. (2012) who found that the heaviest fennel herb fresh
and dry weights were obtained by 200 pp m salicylic acids. Decreasing salicy lic acid to 100 pp m (S1) increased the root
growth characters including root length and root fresh weight as well as shoots dry weight when compared with untreated
plants. Aftab et al. (2010) found that increasing SA brought about a significant growth increase, where the highest
significant increase in shoot dry matter in Artemisia annua L was recorded by 1 mM SA. The role of salicylic acid was
explained by EL-Tayeb (2005) and Hayat et al. (2005) on growth that promotes cell d ivision and enlargement and
Shakirova et al. (2003) who reported that its regulatory effect on cell growth and division is due to its influence on other

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Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

plant hormones, auxin, cytokinins and ABA balances and increased the growth characters. While the combination P1S2
was showed the highest significant value in shoots fresh weight and shoots height.
The combination effect of putrescine and salicylic as well as irrigation treatments on root growth characters
revealed that the twice application of put at 1 ppm (P2) enhanced root growth in plants irrigated every 40 days, and
enhanced shoot growth in plants irrigated every 30 days which could be attributed to its regulatory role in pro moting
productivity and improving plant growth and development due to its effects on cell division, expansion and differentiation
as well as regulates stomatal responses by reducing their aperture and inducing closure. In addition, Hussein et al. (2006)
on pea found that foliar application of putrescine at 2 ppm increased shoot fresh and dry weights under salt stress.
El-Bassiouny and Bekheta (2005) found that Put modulates ABA biosynthesis and stimulated growth by increasing auxin,
gibberellins and cytokinins, which accompanied by ABA inhibitors content and activity reduction in response to abiotic
stress. Application of (S2) resulted in the highest root characters of the plant that irrigated every 20 days. P2S2 t reatment
showed the highest value of root characters in the plants irrigated every 30 days, while P1 showed the highest record in
those irrigated every 40 days. There is no significant effect on the shoot height, fresh and dry weights of shoot which
irrigated every 20 days. On the other hand, it was found the application of putrescine P1 recorded the highest significant
shoot height, fresh and dry weights. However, S2 and P1S2 showed the highest record in shoot height and fresh weight in
40 days irrigated plants. It was found that P1 and S2 application enhanced the all shoot growth characters and root dry
weight than P2 and S1, respectively. On the other hand, the application of S1 was enhanced the root length and root fresh
weight than S2.
Chemical Composition
The mean values of some chemical parameters of A jwain as affected by irrigation, Putrescine and/or salicylic acid
during vegetative stage are shown in Table 3. It was found the reduction in irrigation times significantly increased the total
sugars, total soluble phenols, and free amino acids and significantly decreased chlorophyll b. No significant differences
were found among irrigation treatments in chlorophyll a, total chlorophyll and total carotenoids. These results were
supported by Bernstein (1961) and (Greenway and Munns (1980))who mentioned that the solute accumulation required for
the osmotic adjustment of enlarging cells, that was an indicator of the osmo -protectants levels in stressed plants, which
plays an important role in osmotic adjustment and osmoregulation in non- halophytes. Aly (1987) supposed that the
increase in sugars concentration might be due to the starch and other polysaccharides hydrolysis under water stress
conditions. In addition, Azhar et al. (2011) added that the total phenolic contents increased significantly with increasing
drought stress levels, Whereas they found that the highest total phenolic contents were observed at 60% field capacity
(f.c.), followed by 80% f.c. and 100% f.c.. in Trachyspermu m ammi L. plants which suggests that Desi Ajwain can be
cultivated in areas prone to drought. Chen et al. (2011)found that phenolic contents (rosmarinic acid, ursolic acid and
oleanolic acid) o f Prunella vulgaris L. plants increased under drought stress. Azhar et al. (2011) mentioned that the highest
values of chlorophyll a in Desi Ajwain as well as chlorophyll b and total chlorophyll contents in Withania somnifera were
found in plants growing at 60% field capacity, fo llo wed by plants at 100% and 80% F.C. Also, Desi A jwain has the
potential to maintain a h igh photosynthetic rate under water stress and also the chlorophyll increases. Matthews et al.
(1990) reported the reduction in photosynthetic activity due to the stomatal conductance and water uptake reduction which
cause photosynthetic adaptability.
Regarding the mean effect of both putrescine and salicylic acid, it was found that the twice putrescine application
Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

67

at 1 ppm concentration (P2), significantly increased the sugars, whereas, sugars decreased with once putrescine application
(P1). These results supported by Abdel Aziz et al. (2009) who pointed out that the application of Put on wheat plants
resulted in an increase in the total carbohydrates content which may be attributed to photosynthetic process efficiency and
net assimilation of leaf CO2 increase. Talaat and Balbaa (2010) reported that foliar spraying of PAs significantly increased
total sugar concentration in sweet basil. The free amino acids were significantly increased with putrescine applied,
once and twice, than the untreated plants. It is found that the application of putrescine involved in important biological
process, e.g. ionic balance and DNA, RNA and protein synthesis. Similar suggestions were reported by Hanafy Ahmed et
al. (2002) on Myrtus communis plants. Application of Put once at 1 pp m (P1) leads to increase in total soluble phenols. Das
et al. (1990) added that phenol accumulation could be a cellu lar adaptive mechanism for scavenging oxygen free radicals
during stress, which preventing sub-cellular damages. In addition, Hanafy Ahmed (1997) found that the higher levels of
total soluble phenols and total free amino acids concentration might be due to the increase in the metabolic activity to
synthesis shikimic acid. Application of salicy lic acid at concentration of 100 pp m and 150 pp m were similar to the
untreated plants in total sugars, while the higher concentration (S2) increased the free amino acids values.
The obtained results were supported by Ghasemzadeh and Jaafar (2012) who mentioned that spraying 105 M, SA
enhanced total soluble carbohydrate production.
Table 3: Total Soluble Sugars, Total Sol uble Phenols, Free Amino Aci ds of Ajwain Herb as
Affected by Irrigati on, Putrescine and Salicylic Aci ds and their Interactions
During Vegetati ve Stage, (Combi ned of Seasons 2011-2012)
Character
Treatment
Control
P1
P2
S1
S2
P1S1
P1S2
P2S1
P2S2
Mean A
LSD

Total soluble sugars (mg/g.f.w.)

Mean
Irrigation Times
(B)
7
5
3
14.54

25.13

10.67

16.78

12.83

17.14

13.27

14.41

10.83

17.76

27.74

18.78

11.97

19.47

18.56

16.67

16.78

20.07

13.16

16.67

10.70

12.29

17.39

13.46

12.90

15.04

10.94

12.96

15.56

22.92

24.35

20.95

14.39

22.69

19.53

18.87

13.39

19.17

17.29

A= 2.96

B= 5.12

AB = ns

Total soluble phenols

Mean
Irrigation Times
(B)
7
5
3
1.01
1.43
1.18
1.10
1.07
1.51
1.20
1.03
1.14
1.35
1.15
0.96
1.07
1.23
1.11
1.04
1.12
1.38
1.16
0.99
1.03
1.42
1.13
0.93
0.95
1.53
1.16
0.99
1.21
1.11
1.17
1.21
1.33
1.35
1.25
1.08
1.10
1.37
1.04
B= ns
AB= 0.28
A= 0.1

Free Amino Acids

Irrigation Times
Mean (B)
7
5
3
4.04
8.79
5.59
3.93
8.67
10.45
7.19
2.44
4.28
13.92
7.05
2.96
7.55
6.22
5.79
3.61
10.52
7.88
7.40
3.79
3.58
5.56
4.15
3.31
5.15
7.38
5.94
5.27
3.42
10.52
6.80
6.46
2.91
7.22
5.46
6.24
5.57
8.66
4.22
A=0.45
B= 0.78 AB= 1.36

Our results on phenols were supported by Ghasemzadeh and Jaafar (2013) who worked on ginger as well as
Ibrahim and Jaafar (2011) who worked on Labisia pumila. They pointed out that the increase in total phenols content might
be due to the increase in total soluble sugars production in the leaves. On the other hand, Nicholson, RL Hammersch midt
(1992) reported that SA, at low concentration (105 M), inhibits the PAL activity, which is the key enzyme in phenolic
synthetase and stimulates chalcone synthase activity (CS), which is the key enzyme in flavonoids synthetase.
In addition, Chen et al. (1993) added that SA inhibits catalase activity which leading to H 2 O2 increased, which in turn
induces PAL gene expression and phenolic compounds synthesis.
Regarding chlorophyll, chlorophyll a & total chlorophylls were significantly increased and chlorophyll b & total
carotenoids were decreased with twice putrescine application at 1 ppm concentration (P2) as shown in Table 4. P1S2 and
P2S2 significantly increased the chlorophyll b and total carotenoids and significantly decreased chlorophyll a and total
chlorophyll. In this respect, there is an inversely relation between chlorophyll b and total carotenoids with chlorophyll a
and total chlorophyll might indicate the plants suffered by the stress which affected chlorophyll a reduction and led to its

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68

Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

degradation to chlorophyll b, and subsequently chlorophyll b concentration has increased. Likewise, the increases in total
carotenoids supports that the plants which were treated by these treatments might be under stress. In addition,
Hanafy Ahmed et al. (2002) worked on Myrtus communas, and reported that the increase in chlorophyll concentration
influenced by high Mg +2 content which accumulated by the enhancing effect of putrescine. Krishnamurthy (1991) found
that putrescine inhibited Na and Cl uptake and increased K, Ca and Mg accumu lation in leaves, which enhance chlorophyll
formation. However, Abdel A ziz et al. (2009) added that put sprayed at 100 pp m on g ladiolus plants significant ly
increased chlorophyll a, b, a + b and carotenoids in both seasons. Ma et al. (1996) mentioned the effect of PAs in inhibiting
chlorophyll degradation may be related to the inhibition of pero xidase activity. On the other hand, the treatment S2
recorded the lowest significant one in chlorophyll a, b and total chlorophylls as well as total carotenoids. Fariduddin et al.
(2003) reported that spraying plants with salicylic acid at 10-2 M enhanced the net photosynthetic rate, internal CO2
concentration and water use efficiency in Brassica juncea. Moharekar et al. (2003) found that salicylic acid activated the
synthesis of carotenoids and xanthophylls. In addition, Noreen et al. (2011) added that the contents of photosynthetic
pigments were slightly imp roved with SA application at 200 and 300 mg L-1 . Khodary (2004) suggested that this increase
is related to salicylic stimu latory effects on RuBisCO activ ity.
Table 4: Chl orophyll a, b and Total Chl orophyll as well as Total Carotenoi ds of Ajwain Herb
as Affected by Irrigation, Putrescine and Salicylic Aci ds and their Interactions
During Vegetati ve Stage (Combined of Seasons 2011-2012)

Considering the interaction effect of both putrescine and salicylic acids on plants under drought stress, revealed
that total soluble phenols were recorded the highest significantly value under drought stress. Whereas, the application of
putrescine at 1 ppm (P1) and salicylic acid at 150 ppm (S2) and their co mbination (P1S2) recorded the highest significant
value the total soluble phenols. Free amino acids were increased in 40 days irrigated plants. Putrescine application at
concentration of 1 ppm applied once and twice, significantly increased and increased the free amino acids, respectively.

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

69

The combination treatment (P2S1) increased free amino acids. In this respect, Khan et al. (2000) mentioned that amino
acid must be accumulated to high levels to create an osmotic potential gradient to facilitate inward movement of water.
Plants produce a variety of proteins under biotic and abiotic stresses. SA is known to induce the production of these
proteins which contribute to enhanced plant resistance to salinizat ion as mentioned by Kang and Saltveit (2002).
Akhtar et al. (2013)added that the application of SA at 100 mg L- on maize conducted under saline conditions found to
increases protein and amino acids and EL-Tayeb (2005) on barely Th is increase in amino acids in plant organs is due to the
breakdown of protein and as a result, reduction in protein and growth has been observed (Hussein et al. (2007) and as a
result of decrease in the synthesis due to reduction of NRA, NiRA and substrate. Concerning chlorophyll concentration, the
data presented in table 3 reveal that chlorophyll a, and total chlorophyll were significantly increased and recorded the
highest values under application of P2 in p lants irrigated every 20 days (non -drought). While those irrigated every 40 days,
the severe drought treated by P2 recorded the highest chlo rophyll a and total chlorophyll compared with those plants
supplied by the same irrigation level. On the other hand, the application of the salicylic acid under all irrigation treatmen ts
decreased the chlorophyll a, and total chlorophyll. Ho wever, there we re no significant differences found among treatment
in chlororphyll b and total caretoniods.
There was harmony between our results and those mentioned by Hanafy Ahmed et al. (2002) who suggested that,
the cell exposed to osmotic stress lead to redirect carbon flow to osmoregula t ion by accumulat ion of variety of co mmon
solutes including sugars, amino acids, organic acids and ions (especially K) and another metabolically protective osmolytes
leading to water retention and Carpita et al. (1990) as well as Taiz and Zeiger (1991).
Essential Oil
Table 5: Essenti al Oil% , Oil Content per Plant and Oil Yiel d per Feddan of Ajwain Herb as
Affected by Irrigati on, Putrescine and Salicylic Aci ds and their Interactions During
Vegetati ve Stage (Combined of Seasons, 2011-2012)
Character
Treatment
Control
P1
P2
S1
S2
P1S1
P1S2
P2S1
P2S2
Mean A
LSD

Essential oil percent

Mean
Irrigation Times
(B)
7
5
3
0.14

0.17

0.15

0.15

0.14

0.10

0.16

0.13

0.13

0.14

0.15

0.14

0.14

0.17

0.19

0.17

0.16

0.17

0.19

0.17

0.14

0.17

0.17

0.16

0.16

0.13

0.13

0.14

0.17

0.16

0.18

0.17

0.08

0.19

0.12

0.13

0.14

0.16

0.16

Oil content per plant (ml/plant)

Mean
Irrigation Times
(B)
7
5
3
0.09
0.04
0.12
0.23
0.10
0.04
0.07
0.07
0.05
0.03
0.06
0.10
0.10
0.05
0.11
0.19
0.09
0.04
0.12
0.22
0.08
0.03
0.05
0.04
0.08
0.03
0.09
0.15
0.05
0.04
0.07
0.12
0.09
0.03
0.06
0.07
0.08
0.04
0.13
A= 0.005 B= 0.009 AB= 0.02

Oil yield per Feddan (kg/fed)

Irrigation Times
Mean (B)
7
5
3
9.02
1.78
4.74
3.43
2.96
1.43
2.80
4.00
3.99
1.22
2.39
1.95
7.56
2.03
4.57
4.11
8.93
1.67
4.68
3.44
1.43
1.25
1.91
3.04
6.03
1.13
3.41
3.07
4.79
1.60
2.77
1.93
2.63
1.16
2.41
3.44
5.26
1.48
3.16
A= 0.09
AB= 0.27
B= 0.16

The effect of irrigation, putrescine and/or salicylic acid t reatments on essential oil are shown in Tab le (5).
Increasing irrigation intervals increased the essential oil percent. Plants irrigated either 5 or 3 times resu lted in the highest
oil percent, whereas the opposite was true with oil content per plant, since plants irrigated seven times contained the
highest oil content per plant and oil yield per Feddan. In this respect, Mohamed and Abdu (2004) mentioned that under
water stress, the percentage of volatile oils in parsley and fennel increased. Moreover, Vah idipour et al. (2013) worked on
Ajwain, found that increasing irrigation intervals at 14 days recorded the highest essential oil percentage, whereas, the
lowest was gained from the irrigation of 7 days irrigation, but they was not statistically significant . Also, Bahreininejad et
al. (2013) who mentioned that water stress increased essential o il content, but decreased essential oil y ield o f T. daenensis.

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70

Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

In addition, Charles et al. (1990) wo rked on peppermint and found that the slight increase in the essential oil
percentage in treatment of 14 days irrigation that plant is exposed to drought stress, can be because of the greater density of
the essential oil secreting glands in consequence of the lower leaf surface due to stress, causes the essential oil is more
concentrated.
Regarding the mean effect of the salicylic acids and putrescine, results revealed that the applica tion of salicylic
acid significantly increased essential oil percent and did not show considerable changes in oil content and oil yield
comparing with control plants. Application of putrescine treatments decreased essential oil percent, oil content per pla nt
and oil yield per Feddan comparing to control plants . Combination between salicylic acid and putrescine varied in their
effect on the essential oil percent comparing to control plants. Treatments of P1S1 and P2S1 increased oil percent, where
PIS2 and P2S2 decreased oil percent decreased oil percent comparing to control plants. All the combination treatments
decreased oil content and oil yield co mparing to control plants. It is clear that salicylic acid treatments increased the
tolerant of Ajwain plants to drought stress and resulted in the highest oil yield comparing to other treatments.
In this respect, Sangwan et al. (2001) reported that the application of growth regulators which affect oil components might
be due to their effects on enzymat ic pathways of terpenoid biosynthesis. In addition, Sharafzadeh and Zare (2011)
mentioned that growth regulators can influence essential oil production through its effects on plant growth, essential oil
biosynthesis and the number of oil storage structures.
Concerning the effect of co mbination of salicylic acid and Putrescine on essential oil of Ajwain p lant herb
subjected to drought stress, results revealed that the highest significant value of essential oil % was recorded in plants
irrigated every 40 days and treated with S1, S2. In addit ion, the P2S1 was the most effective treat ment in plants irrigated
every 20 days and P2S2 in those irrigated every 30 days. It was found that lowest irrigated plants contained the highest
essential oil percent.
In addition, concerning the oil content per plant, it was found that control and S2 showed the highest significant
oil content in plants irrigated every 20 days. Whereas, those irrigated every 40 days and treated with S1 (100 ppm)
recorded the highest oil content per plant.
Regarding the oil y ield per Feddan, it was found that control and S2 treat ments recorded the highest significant
value of oil yield per Feddan. While in the severe stressed plants, irrigated every 40 days, S1 was found to be the best
treatment among severe drought stressed plants. Finally, it was found fro m the previous that salicylic acid was enhancing
the oil%, oil per plant and per Feddan, at the vegetative growth, under non -stressed and severe drought stressed plants.
The results were in harmony with Azhar et al. (2011) who mentioned that Trachyspermum ammi L. plants decreased
vegetative biomass accumulation but enhanced their phenolic contents under drought stress and this suggests that desi
Ajwain can be cultivated in areas prone to drought. The reduction in plant growth was less than 50% and plants have the
ability to cope with stress by generating secondary metabolites including phenols. And Terzi et al. (2010) added that the
main importance of phenol compounds is related to the inhibitory potential of their hydroxy l group for free radicals.
Hence, an elevated level of this co mpound could be considered an index of enhanced anti-oxidative activ ity of essential oil.
Essential Oil Components
The main constituents of the essential oil o f Ajwain fresh herb as identified by GLC for the different treatments
are shown in Tables 5. All treat ments showed -terpinene as the main constitute of the essential oil of the Ajwain herb.

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

71

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

Cy mene found to be the second main component of the essential oil of Ajwain herb in all different irrigated plants.
Thymol ranged from 0.40 to 1.09 % fro m all identified co mponents in the plants irrigated every 20 days and ranged from
0.26 to 1.83% in the oil of the plants irrigated every 30 days. It ranged fro m 0.27 to 2.23% in the essential oil of A jwain
vegetative herb irrigated every 40 days. These results indicated t hat drought stress enhanced and
-

,S b

-pinene,

d - terpinolene, and Thymol. Azhar et al. (2011) reported that cultivation of a medicinal plant like

Desi Ajwain (Trachyspermum ammi) in water-deficient areas would increase its defense system and the level of active
compounds. The obtained results indicated that Thymol increased with increasing drought stress. Likewise, Bahrein inejad
et al. (2013) demonstrated that percentage of Thymol increased under moderate and severe water stress in the essential oil
fro m T. daenensis. On the contrary, Alavi-saman i et al. (2013) mentioned that the amount of Thymol decreased under
water deficit stress. The trea m
4 d y ,r p

v y,

P2, P2S1
d

d P2S2 r
r

rd d
y

-terpinene in plants irrigated every 20, 30 and

d

-terpinene under the increase of

the water stress, Whereas, P2S2 recorded the highest cymene in the plants irrigated every 20 and 40 days.
However, it recorded the highest value in plants irrigated every 30 days treated by P2S1. In addit ion, Thymo l was
increased with all applied treatments in the plants irrigated every 20 days except with S1 and P2S2, whereas, P2 the
increased Thymol by 109% than control.
Finally, it was found that the majority of the components of the oil were increased in the plants irrigated every 20
and 40 days by the treatment P1S2, whereas the oil components in the plants irrigated every 30 days were increased by the
treatment P2S2. Gharib and EL-Lateef (2006) found that SA application significantly increased carvacrol, -a-thujene,
a-pinene and p-cymene, and decreased Thymol and b-caryophyllene contents in the thyme essential oil. Also, SA may
convert Thymol to its isomer (carvacrol), and is thought to play an important signaling role in the activation of various
plant defense responses, such as the biosynthesis of special secondary metabolites, which function as phytoalexins in
plants, the results are in agreement with Kang et al. (2004)). Lamb and Dixon (1997) and Sh i et al. (2006) added that the
induction mechanism of plant defense is generally thought to be related to the elevation of H 2 O2 and other reactive oxygen
species (ROS), which can then serve as second messengers in the defens e signaling pathway.
El-Ghorab et al. (2006) added that carnation (Dianthus caryophyllus) flower have variations in the essential oil
components as a result of treating with different concentrations of and/or putrescine and their synergetic, it could be due to
the increase in the terpene biosynthesis and /or might be due to an indirect effect on carbohydrate metabolis m. Also, the
author found that the percentage of hexadecanoic acid co mponent was highly increased as a result of treating with 200 pp m
putrescine and the maximu m decrease in hexadecanoic acid was obtained fro m application of 400-pp m putrescine.
Table 6: The Components of the Ajwain Herb Oil as Affected by Irrigati on, Putrescine and
Salicylic Acids and their Interacti ons during Vegetati ve Stage
(Combined Of Seasons, 2011-2012)
Oil
components

Contr
ol

P1

-piene

0.96

0.96

-Thujene

2.80

2.83

Sabiene

4.23

4.55

B-my rceme

6.51

6.62

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7 Irrigated Ti mes (Every 20 Days)

P1S
P2
S1
S2
P1S1
2
1.0
0.84
0.99
0.94
0.97
0
2.7
2.50
3.00
2.75
2.85
2
4.4
4.05
4.54
4.06
4.55
3
5.95 5.1
6.20
5.84
6.54

P2S
1
1.0
1
2.9
8
4.4
4
6.8

P2S2
0.60
1.67
3.78
4.70
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72

Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

D-Limonene

1.77

1.90

1.68

-terpinene

47.96

26.5
4
15.5
1

48.6
0
15.0
0

Cy mene

15.63

terpinolene

3.23

3.36

3.67

Thymol

0.52

1.09

0.91

Carvacol

0.45

0.88

0.63

Oil
components
-piene
-Thujene
Sabiene
B-my rceme
D-Limonene

1.01
3.12
4.41
6.01
1.55

-terpinene

29.59

Cy mene

13.78

terpinolene
Thymol
Carvacol
Oil
components
-piene
-Thujene
Sabiene
B-my rceme
D-Limonene

1.86

1.88

1.87

45.81

34.3
4

9.34

8.47

3.54

2.58

2.75

0.88

0.91

0.92

0.82

0.97

0.71

27.7
7
14.2
3

0
1.8
5
47.
49
13.
32
3.8
3
0.8
6
0.5
7

2.19
21.86
18.84
5.62
0.40
1.10

5 Irrigated Ti mes (Every 30 Days)

0.77
2.22
4.05
4.86
1.93
45.0
1
17.7
5

0.71
2.09
3.91
5.92
1.75
28.9
6
17.4
6

3.99

5.84

1.01
0.34

0.26
0.80

1.00
3.01
4.20
6.30
1.72

0.94
2.77
4.32
6.11
1.65

0.89
2.65
4.26
5.95
1.71

0.64
1.76
3.72
6.48
1.90

0.52
1.62
2.56
4.02
1.40
54.6
1
20.7
8

1.11
3.14
4.76
6.76
2.45
44.6
1
15.1
0

26.65

48.40

46.96

45.61

15.16

16.21

9.33

15.50

3.14

2.98

2.63

3.09

3.34

1.76

3.44

0.90
0.75

0.94
0.67

0.45
0.70

0.31
0.53

1.83
0.74

0.67
0.50

0.81
0.93

0.94
2.67
4.13
7.46
2.02
30.6
1
13.6
6

0.24
0.74
1.57
3.04
1.03
53.5
3
25.5
9

3 Irrigated Ti mes (Every 40 Days)

0.71
2.06
3.47
5.21
1.65

0.68
2.05
3.54
5.78
1.65
47.7
9
15.5
8

0.59
1.81
2.95
4.51
1.34
27.5
2
21.5
6

3.93

3.69

1.22
0.62

2.23
0.71

-terpinene

25.09

Cy mene

17.18

terpinolene
Thymol
Carvacol

4
2.2
5
43.
96
15.
94
3.8
5
0.4
0
1.1
8

Impact Factor (JCC): 4.3594

0.96
3.02
4.19
6.70
1.65

0.93
2.44
3.95
5.72
2.26

0.95
2.75
3.93
6.52
1.86

0.93
2.71
4.39
6.33
2.02

48.56

24.69

45.67

23.32

13.31

16.91

15.43

15.46

2.51

3.47

2.97

3.23

5.59

3.92

2.23

1.96
1.18

2.03
0.76

1.17
0.86

1.20
0.76

0.27
0.64

2.81
0.89

0.36
0.50

Index Copernicus Value (ICV): 3.0

Effe ct of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Ammi) at Vegetative Stage Grown Under Drought Stress

73

Table 7: The Oxygenated and Non-Oxygenated Components in the Ajwain Herb Oil as
Affected by Irrigati on, Putrescine and Salicylic Aci ds and their
Interactions during Vegetati ve Stage (Combined Of Seasons, 2011-2012)
Oxygenated compounds
Treatment

Irrigation Times

None-oxgygenated compounds
Mean

Control

0.97

1.35

1.84

P1

1.97

1.06

2.94

P2

1.54

1.65

S1

1.58

S2

1.70

P1S1

Irrigation Times

Mean

1.39

83.09

63.46

59.30

68.62

1.99

62.27

82.43

80.76

75.15

3.14

2.11

82.29

63.94

62.79

69.67

1.61

2.79

1.99

79.29

61.02

81.86

74.06

1.15

2.03

1.63

62.13

83.03

59.87

68.34

1.88

0.84

1.96

1.56

73.20

74.84

80.34

76.13

P1S2

1.63

2.57

0.91

1.70

62.34

78.95

60.75

67.35

P2S1

1.43

1.17

3.70

2.10

81.72

87.27

65.41

78.13

P2S2

1.50

1.74

0.86

1.37

59.26

81.37

87.97

76.20

Concerning the oxygenated and none-oxygenated compounds, the mean values of the irrigation treatments
indicated that the oxygenated compounds increased with increasing irrigation intervals. Ajwain plants irrigated every 40
days showed the highest values of oxygenated compounds, where the lowest values were recorded in those irrigated every
30 days. On the other hand, the reverse was found with the non -o xygenated compounds. The mean effect of P2 showed the
highest value of oxygenated compounds, whereas, the lowest values were recorded by P2S2. On the other hand, P2S1
recorded the highest percent of the non-oxygenated compounds, whereas, the lowest values were recorded with P1S2
treatment. The interaction effect of both the salicylic and putrescine on the oxygenated and non-oxygenated compounds of
plants subjected to drought stress revealed that P1, P1S2 and P2S1 recorded the highest oxygenated compounds in plants
irrigated every 20, 30 and 40 days, respectively. Whereas, the lowest value was recorded by the control, P1S1 and P2S2 in
plants irrigated every 20, 30 and 40 days, respectively. On the other hand, the highest values of non-oxygenated
compounds were recorded with control, P2S1 and P2S2 in the plants irrigated every 20, 30 and 40 days. Whereas, P2S2,
S1 and the control showed the lowest values of non-oxygenated compounds in the plants irrigated every 20, 30 and 40
days, respectively. Lawrence (1988) reported that oxygenated compounds ranged from 55.30% to 83.12% in plants treated
with 150 mg/ l and 25 mg/ l Put, respectively. This may be attributed to that the biosynthetic pathways of the main
compounds in the basil oil are either independently or simu ltaneously fro m shikimic acid or mevalonic acid. Also, the
author added that stress condition may alter the biosynthetic process to the advantage of methyl chavicol fro m shikimic
acid / or mevalonic acid, since green ruffles basil varieties are dual biosynthetic pathways.
Yiel d
Table 8: Herb Fresh (ton/fed) and Dry Yiel d (ton/fed) of Ajwai n Pl ant as Affected by Irrigation, Putrescine,
Salicylic Acids and their Interacti ons, during Vegetati ve Stage (Combined of Seasons 2011-2012)
Character
Treatment
Control
P1
P2
S1
S2
P1S1
P1S2
P2S1
P2S2
Mean A
LSD

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Herb Fresh Yield (ton/Feddan)

Irrigation Times
7
6.55
3.54
2.99
5.40
5.58
2.70
3.77
2.76
3.29
4.07
A=0.62

5
1.99
4.14
1.39
2.37
2.00
1.79
2.45
1.23
1.78
2.13
B=1.07

3
1.01
0.87
0.84
1.07
0.88
0.74
0.85
0.88
0.94
0.90
AB=1.86

Herb Dry Yield (ton/Feddan)

Mean (B)
3.18
2.85
1.74
2.95
2.82
1.74
2.36
1.62
2.01

Irrigation Times
7
0.79
0.44
0.38
0.65
0.69
0.33
0.46
0.35
0.41
0.50
A=0.04

5
0.26
0.55
0.19
0.31
0.27
0.24
0.32
0.17
0.24
0.28
B=0.09

3
0.14
0.12
0.12
0.15
0.13
0.11
0.12
0.13
0.14
0.13
AB=0.16

Mean (B)
0.39
0.37
0.23
0.37
0.36
0.23
0.30
0.21
0.26

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74

Zeid, F. A, Omer, E. A, Amin, A. Y & Hanafy, Shaimaa A. H.

Considering the mean effect of irrigation treatments on the herb fresh and dry yield, seed, it was found that
increasing the drought stress decreased the yield of herb fresh and dry weights at the vegetative growth stage.
Regarding the effect o f salicy lic acid and Putrescine on yield of A jwain plant, results revealed that control without
both putrescine and salicylic acid recorded the highest significant values of herb fresh and dry yield. Whereas application
of putrescine once at concentration of 1 pp m (P1) and salicy lic acid at concentration of 100 ppm (S1) and 150 ppm (S2)
ppm as well as the combination (P1S2) were recorded the high values of herb fresh and dry yield per Feddan,
In this respect, Crisosto et al. (1988) reported that with the application of putrescine at 10-3 M the fru it set, crop
density and yield efficiency were increased. In fact, Putrescine at 10-3 M increased fruit weight, diameter and seed content.
Thus, higher seed number per fru it could explain the trend toward increase in fruit size fro m putrescine.
Regarding the effect of salicylic acid and Putrescine on the Ajwain plant grown under drought stress, it was found
that the highest herb fresh and dry yield per Feddan was recorded with plants irrigated every 20 days without putrescine
and salicylic acids application. While the herb fresh and dry yields per Feddan were increased by the application of S2
(150 pp m) and P1 (1 pp m once) which recorded the highest values under moderate irrigation, plants which irrigated every
30 days. The highest herb fresh and dry yield were recorded with plants treated with S2 (150 pp m) and irrigated every 20
days.

CONCLUSIONS
Drought stress significantly reduced all roots and shoot morphological growth characters, as well as shoots: root
ratio and chlorophyll b of Ajwain plant. While total sugars, total soluble phenols, and free amino acids in shoots and roots
were significantly increased. The essential oil % increased by irrigation interval every 40 days, whereas, oil content per
plant increased by irrigation every 30 days. It was found that P1 and S2 application enhanced the all shoot growth
characters and root dry weight than P2 and S1, respectively. On the other hand, the application of S1 was enhanced the root
length and root fresh weight than S2. The total soluble sugars were increased by P2 and P2S1 in plants irrigated every 40
days. In addition, P1, S2 and P1S2 recorded the highest total soluble phenols. Furthermore, P1, P2 as well as P2S1
significantly increased free amino acids. Chlorophyll a and total chlorophyll were significantly increased with putrescine
twice applicat ion (P2). On the other hand, this treatment decreased the chlorophyll b and total carotenoids, whereas the
chlorophyll b and total carotenoids were significantly increased with P1S2 and P2S2 treat ments. The oil % and oil content
per plant as well as oil yield per Feddan significantly increased by S1 and S2 in those irrigated every 40 days. It was found
terpiene was the main constituent and followed by P cymeme and were increased under water stress . P2 increased
Thymol by 109% than control. In addition, P2S1 and P2S2 were recorded the highest oxygenated and non -oxygenated
compounds in plants irrigated every 40 days. Plants irrigated every 30 and 40 days and treated by S2 and S1 recorded the
highest values of herb fresh and dry yields, respectively.

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