effect of putrescine and salicylic acid on ajwain plant (trachyspermum amm.) at vegetative stage...

8/10/2019 Effect of Putrescine and Salicylic Acid on Ajwain Plant (Trachyspermum Amm.) at Vegetative Stage Under Drought …

http://slidepdf.com/reader/full/effect-of-putrescine-and-salicylic-acid-on-ajwain-plant-trachyspermum-amm 1/20

www.tjprc.org [email protected]

International Journal of AgriculturalScience and Research (IJASR)ISSN(P): 2250-0057; ISSN(E): 2321-0087

Vol. 4, Issue 6, Dec 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, 4Plant Physiology Section, Department of Agricultural Botany, Faculty of Agricu lture,

Cairo University, Giza, Egypt

2Department 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 s tage, grown under drought s tress. 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 ppm, S1), (150 ppm, S2) and their combinations (P1S1,

P1S2, P2S1, P2S2), which were applied at 45 and 60 days from 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 treatments. 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 . α – terpiene

was found to be the main constituent and followed by p – cymeme. 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, Pigments Composition, Yield, Essential Oil, GC-MS

INTRODUCTION

Desi Ajwain (Trachyspermum ammi L.) is an aromatic herb. It belongs to family Apiaceae (Umbelliferae).



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

Impact Factor (JCC): 4.3594 Index Copernicus Value (ICV): 3.0

This plant has several medical 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, which located between the cells, where they act as hormones, regulators and catalysts in the

vegetable metabolis m. They s eem to help the plant to adapt to its environment and are consequently produced in higher

quantities when plants meet extreme conditions , biotic and abiotic st resses such as drought danger.

The water status in Egypt has a critical situation. It is facing an annual water deficit of around 7 billion m3.

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 from 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 a rid and semi-arid regions.

Drought stress alters the plant photosynthesis, respiration, translocation, ion uptake, carbohydrates, 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 plants, like artemisinin 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 plant like Desi Ajwain (Trachyspermum ammi) in water-deficient areas would

increase its defense system and the level of active compounds.

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 implicated 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 (Spm) have been shown to be involved in a variety of plant growth and developmental processes, including cell

division, vascular differentiation, root initiation, shoot formation, flower initiation and development, fruit ripening and

senescence and embryo formation 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 role in the growth and development of plants. The axial role of salicylic acid in translating

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, fruit yield, glycolysis, 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.



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 e xperiments were carried out during the two successive seasons 2011/2012 and 20 12/2013 at the

Agricultural Research and Experimental Station, Faculty of Agriculture, Cairo University, Giza, Egypt. The study

investigated the effect of foliar applications of salicylic ac id and putrescine on Ajwain plant subjected to a drought stress.The Desi Ajwain (Trachspermum 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 des igned 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 from planting) DFP. Each plot consisted of three rows (3 m length and 60

cm width). All the agronomic recommended practices for Apiaceae family were applied. The plants were fertilized by 120

kg calcium super phosphate / Fadden (15.5% P2O5) and 250 kg/fed ammonium sulphate (20.5 %N) and 120 kg/fed of

potassium sulphate 48%. A ll ferti lizer was added at 30, 45 and 60 days from sowing. So il samples were randomly taken

each year before cultivation 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 for both seasons of the

soil are shown in Tab le 1.

Table 1: Mechanical and Chemical Analysis of the Soil Experimental Site

Mechanical Analysis

Coarse

Sand %Fine Sand % Silt % Clay % Texture Class

1.5 – 6.0 37.0 – 41.2 22.0 – 26.1 31.2 – 35.0 Clay loam

Chemical analysispH EC Soluble Anions (meq/l) Soluble Cations (meq/l)

7.9 2HCO3

- Cl

-1 SO4

- Na

+1 K

+1 Ca

+2 Mg

+2

0.6 2 3.7 3.8 0.2 2 0.3

Mechanical Analysis

Soil Depth (cm)Field Capacity

%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 andchemical 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;

Determination of the Essential Oil (ml/100 g) of each sample 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 treatment 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 multiplication the fresh weight of plant by essential

oil percent. Essential oil yield (L/fed.) was calculated by multiplication the yield of fresh herb (Ton / fed.) for each

treatment by the essential oil percentage for each treatment. The GC-Ms analysis of the essential oil sa mples was carried





out in the first season us ing 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

Chromatographs (THERMO Scientific Corp, USA), coupled with a THERMO mass spectrometer detector (ISQ

Single Quadrupole Mass Spectrometer). The GCMS system was equipped with a TG-WAX MS column (30 m x 0.25 mm

i.d, .25 μm fm k). Ay r rrd m rrr f r f 1. mL/m p

ratio of 1:10 and the following temperature program: 40°C for 1 min; rising at 4.0 C/min to 160°C and held for 6 min;

rising at 6°C/min to 210°C and held for 1 min. The injector and detector were held at 200 and 200°C, respectively.

Dd mp (1:1 x, v/v) f .2 μL f mxr r y jd. M pr r bned 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)

confirmation 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 chlorophylls and total

carotenoids) were determined in leaves samples (mg/g fresh weight) by using dimethyl-formamide according to

Nornai (1982).

Yield: 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 des ign

according to the procedures outlined by Snedecor and Cochran (1980). LSD at 5% level of significance was used to

compare treatments means. All statistical analysis was performed by using analysis of variance technique of

(Mstat-C, 1989) Computer software package.

RESULTS AND DISCUSSIONS

Growth CharactersThe 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 s tudied treatments showed significant effect on

all growth characters of Ajwain plants except the number of branches. I generally, increasing the irrigation intervals

(decreasing the number of irrigation times) decreased all growth 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 recorded 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





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 moisture

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 length, Fresh and Dry Weights of Shoot and Root and Shoot: Root Ratio as well as

Number of Branches of Ajwain pl ant as Affected by Irrigation, Putrescine, Salicylic aci d and their Interactions

during Vegetative Stage (Combined of Seasons 2011-2012 and 2012-2013)Character

7 5 3 7 5 3 7 5 3 7 5 3

Control 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

P1 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

P2 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

S1 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

S2 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

P1S1 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

P1S2 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

P2S1 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

P2S2 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

Mean A 101.64 53.18 22.42 12.44 7.04 3.20 4.61 3.86 3.30 0.57 0.51 0.47

LSD A= 7.86 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

CharacterControl 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

P1 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

P2 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

S1 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

S2 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

P1S1 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

P1S2 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

P2S1 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

P2S2 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

Mean A 38.76 41.80 38.43 15.37 12.87 10.99 21.94 14.02 7.04 22.50 25.38 17.72

LSD 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

Shoot root ratio Number of branchesShoot Height (cm) Root length (cm)

Irrigation TimesMean (B)

Irrigation Times Mean

(B)

Shoot fresh weight (g) Shoot dry weight (g) Root fresh weight (g) Root dry weight (g)

TreatmentIrrigation Times Irrigation TimesMean

(B)

Mean

(B)

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-Toha my et al. (2008)) reported that putrescine spraying increased all growth parameters. Application of salicylic acid at concentration of

150 ppm (S2) significantly increased the shoot growth characters, the fresh and dry weights 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 ppm salicylic acids. Decreasing salicylic acid to 100 ppm (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 division 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





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 promoting

productivity and improving plant growth and development due to its effects on cell division, e xpansion and differentiation

as well as regulates stomatal respons es 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 adjus tment 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 Trachyspermum 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 ac id) of Prunella vulgaris L. plants increased under drought stress. Azhar et al. (2011) mentioned that the highest

values of chlorophyll a in Desi A jwain as well as chlorophyll b and total chlorophyll contents in Withania somnifera were

found in plants growing at 60% field capacity, followed by plants at 100% and 80% F.C. Also, Desi Ajwain has the

potential to maintain a high photosynthetic rate under water st ress 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





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 suggest ions were reported by Hanafy Ahmed et

al. (2002) on Myrtus communis plants. Application of Put once at 1 ppm (P1) leads to increase in total soluble phenols. Das

et al. (1990) added that phenol accumulation could be a cellular 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 salicylic acid at concentration of 100 ppm and 150 ppm 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 10−5

M, SA

enhanced total soluble carbohydrate production.

Table 3: Total Soluble Sugars, Total Soluble Phenols, Free Amino Aci ds of Ajwain Herb as

Affected by Irrigation, Putrescine and Salicylic Acids and their Interactions

During Vegetative Stage, (Combi ned of Seasons 2011-2012)

Character

7 5 3 7 5 3 7 5 3

Control 14.54 25.13 10.67 16.78 1.10 1.01 1.43 1.18 4.04 3.93 8 .79 5.59

P1 12.83 17.14 13.27 14.41 1.03 1.07 1.51 1.20 8.67 2.44 1 0.45 7.19

P2 10.83 17.76 27.74 18.78 0.96 1.14 1.35 1.15 4.28 2.96 1 3.92 7.05

S1 11.97 19.47 18.56 16.67 1.04 1.07 1.23 1.11 7.55 3.61 6 .22 5.79

S2 16.78 20.07 13.16 16.67 0.99 1 .12 1.38 1.16 10.52 3.79 7 .88 7.40

P1S1 10.70 12.29 17.39 13.46 0.93 1.03 1.42 1.13 3.58 3.31 5 .56 4.15

P1S2 12.90 15.04 10.94 12.96 0.99 0.95 1.53 1.16 5.15 5.27 7 .38 5.94

P2S1 15.56 22.92 24.35 20.95 1.21 1.21 1.11 1.17 3.42 6.46 1 0.52 6.80

P2S2 14.39 22.69 19.53 18.87 1.08 1.33 1.35 1.25 2.91 6.24 7 .22 5.46

Mean A 13.39 19.17 17.29 1.04 1.10 1.37 5.57 4.22 8.66

LSD A= 2.96 B= 5.12 AB = ns A= 0.1 B= ns AB= 0.28 A=0.45 B= 0.78 AB= 1.36


Total soluble sugars (mg/g.f.w.) Total soluble phenols Free Amino Acids


(B)

Mean

(B)

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 Hammerschmidt

(1992) reported that SA, at low concentration (10−5

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 2O2 increased, which in turn

induces PAL gene e xpress ion 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





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 accumulation in leaves, which enhance chlorophyl l

formation. However, Abdel Aziz et al. (2009) added that put sprayed at 100 ppm on gladiolus plants significantly

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 peroxidase activity. On the other hand, the treatment S2

recorded the lowest significant one in chlorophyll a, b and total chlorophylls as well as to tal 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 slight ly improved with SA application at 200 and 300 mg L-1. Khodary (2004) suggested that this increase

is related to salicylic stimulatory effects on RuBisCO activity.

Table 4: Chl orophyll a, b and Total Chlorophyll as well as Total Carotenoids of Ajwain Herb

as Affected by Irrigation, Putrescine and Salicylic Acids and their Interactions

During Vegetative 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 combination (P1S2) recorded the highest s ignificant

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.





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 mgL- on maize conducted under saline conditions found to

increases protein and amino acids and EL-Tayeb (2005) on barely This increase in amino acids in plant organs is due to the

breakdown of protein and as a result, reduct ion in protein and growth has been observed (Huss ein 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 plants irrigated every 20 days (non-drought). While those irrigated every 40 days,

the severe drought treated by P2 recorded the highest chlorophyll 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 t reatmen ts

decreased the chlorophyll a, and total chlorophyll. However, 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 osmoregulation by accumulation of variety of common

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).

Ess ential Oil

Table 5: Ess ential Oil% , Oil Content per Plant and Oil Yiel d per Feddan of Ajwain Her b as

Affected by Irrigation, Putrescine and Salicylic Acids and their Interactions During

Vegetati ve Stage (Combined of Seasons, 2011-2012)

Character

7 5 3 7 5 3 7 5 3

Control 0.14 0.17 0.15 0.15 0.23 0.09 0.04 0.12 9.02 3.43 1.78 4.74

P1 0.14 0.10 0.16 0.13 0.07 0.10 0.04 0.07 2.96 4.00 1.43 2.80

P2 0.13 0.14 0.15 0.14 0.10 0.05 0.03 0.06 3.99 1.95 1.22 2.39

S1 0.14 0.17 0.19 0.17 0.19 0.10 0.05 0.11 7.56 4.11 2.03 4.57

S2 0.16 0.17 0.19 0.17 0.22 0.09 0.04 0.12 8.93 3.44 1.67 4.68

P1S1 0.14 0.17 0.17 0.16 0.04 0.08 0.03 0.05 1.43 3.04 1.25 1.91

P1S2 0.16 0.13 0.13 0.14 0.15 0.08 0.03 0.09 6.03 3.07 1.13 3.41

P2S1 0.17 0.16 0.18 0.17 0.12 0.05 0.04 0.07 4.79 1.93 1.60 2.77

P2S2 0.08 0.19 0.12 0.13 0.07 0.09 0.03 0.06 2.63 3.44 1.16 2.41

Mean A 0.14 0.16 0.16 0.13 0.08 0.04 5.26 3.16 1.48

LSD A= 0.005 B= 0.009 AB= 0.02 A= 0.09 B= 0.16 AB= 0.27

Essential oil percent Oil content per plant (ml/plant) Oil yield per Feddan (kg/fed)


(B)

Mean

(B)


The effect of irrigation, putrescine and/or salicylic acid treatments on essential oil are shown in Table (5).

Increasing irrigation intervals increased the ess ential 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, Vahidipour 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, Bahreinine jad et

al. (2013) who mentioned that water stress increased essential oil content, but decreased essential oil yield o f T. daenensis.





In addition, Charles et al. (1990) worked 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 application 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 plant

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 comparing 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 enzymatic 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 s tructures.

Concerning the effect of combination of salicylic acid and Putrescine on essential oil of Ajwain plant 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 addition, the P2S1 was the most effective treatment 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 yield per Feddan, it was found that control and S2 treat ments recorded the highest s ignificant

value of oil yield per Feddan. While in the severe stressed plants, irrigated every 40 days, S1 was found to be the best

treatment a mong severe drought stressed plants. Finally, it was found from the previous that sal icylic acid was enhancing

the oil%, oil per p lant 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 hydroxyl group for free radicals.

Hence, an elevated level of this compound could be considered an index of enhanced anti-oxidative activity of essential oil.

Ess ential Oil Components

The main constituents of the essential oil of Ajwain fresh herb as identified by GLC for the different t reatments

are shown in Tables 5. All treatments showed α-terpinene as the main constitute of the essential oil of the Ajwain herb.





Cymene 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 % from all identified components in the plants irrigated every 20 days and ranged from

0.26 to 1.83% in the oi l of the plants irrigated every 30 days. It ranged from 0.27 to 2.23% in the essential oil of A jwain

vegetative herb irrigated every 40 days. These results indicated that drought stress enhanced an d rd α-pinene,

α- j, Sb d α- 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 s tress. Likewise, Bahreininejad

et al. (2013) demonstrated that percentage of Thymol increased under moderate and severe water stress in the essential oil

from T. daenensis. On the contrary, Alavi-samani et al. (2013) mentioned that the amount of Thymol decreased under

water deficit stress. The tream P2, P2S1 d P2S2 rrdd α-terpinene in plants irrigated every 20, 30 and

4 dy , rpvy, dd r y d d α -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 addition, Thymol 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 Shi et al. (2006) added that the

induction mechanism of plant defense is generally thought to be related to the elevation of H 2O2 and other reactive oxygen

species (ROS), which can then s erve 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 ppm

putrescine and the maximum decrease in hexadecanoic acid was obtained from application of 400-pp m putrescine.

Table 6: The Components of the Ajwain Herb Oil as Affected by Irrigation, Putrescine and

Salicylic Acids and their Interactions during Vegetative Stage

(Combined Of Seasons, 2011-2012)

7 Irrigated Times (Every 20 Days)

Oil

components

Contr

olP1 P2 S1 S2 P1S1

P1S

2

P2S

1P2S2

α-piene 0.96 0.96 0.841.0

00.99 0.94 0.97

1.0

10.60

α-Thujene 2.80 2.83 2.502.7

23.00 2.75 2.85

2.9

81.67

Sabiene 4.23 4.55 4.05

4.4

3 4.54 4.06 4.55

4.4

4 3.78B-myrceme 6.51 6.62 5.95 5.1 6.20 5.84 6.54 6.8 4.70





4 0

D-Limonene 1.77 1.90 1.682.2

51.86 1.88 1.87

1.8

52.19

α-terpinene 47.9626.5

4

48.6

0

43.

96

27.7

745.81

34.3

4

47.

4921.86

Cymene 15.6315.5

115.0

015.94

14.23

9.34 8.4713.32

18.84

α-

terpinolene3.23 3.36 3.67

3.8

53.54 2.58 2.75

3.8

35.62

Thymol 0.52 1.09 0.910.4

00.88 0.91 0.92

0.8

60.40

Carvacol 0.45 0.88 0.631.1

80.82 0.97 0.71

0.5

71.10

Oil

components5 Irrigated Times (Every 30 Days)

α-piene 1.01 0.77 0.71 1.00 0.94 0.89 0.64 0.52 1.11

α-Thujene 3.12 2.22 2.09 3.01 2.77 2.65 1.76 1.62 3.14

Sabiene 4.41 4.05 3.91 4.20 4.32 4.26 3.72 2.56 4.76B-myrceme 6.01 4.86 5.92 6.30 6.11 5.95 6.48 4.02 6.76

D-Limonene 1.55 1.93 1.75 1.72 1.65 1.71 1.90 1.40 2.45


1

28.9

626.65 48.40 46.96 45.61

54.6

1

44.6

1

Cymene 13.7817.7

5

17.4

615.16 16.21 9.33 15.50

20.7

8

15.1

0

α-

terpinolene3.99 5.84 3.14 2.98 2.63 3.09 3.34 1.76 3.44

Thymol 1.01 0.26 0.90 0.94 0.45 0.31 1.83 0.67 0.81

Carvacol 0.34 0.80 0.75 0.67 0.70 0.53 0.74 0.50 0.93

Oil

components3 Irrigate d Times (Every 40 Days)

α-piene 0.71 0.68 0.59 0.96 0.93 0.95 0.93 0.94 0.24α-Thujene 2.06 2.05 1.81 3.02 2.44 2.75 2.71 2.67 0.74

Sabiene 3.47 3.54 2.95 4.19 3.95 3.93 4.39 4.13 1.57

B-myrceme 5.21 5.78 4.51 6.70 5.72 6.52 6.33 7.46 3.04

D-Limonene 1.65 1.65 1.34 1.65 2.26 1.86 2.02 2.02 1.03


9

27.5

248.56 24.69 45.67 23.32

30.6

1

53.5

3

Cymene 17.1815.5

8

21.5

613.31 16.91 15.43 15.46

13.6

6

25.5

9

α-

terpinolene3.93 3.69 2.51 3.47 2.97 3.23 5.59 3.92 2.23

Thymol 1.22 2.23 1.96 2.03 1.17 1.20 0.27 2.81 0.36

Carvacol 0.62 0.71 1.18 0.76 0.86 0.76 0.64 0.89 0.50





Table 7: The Oxygenated and Non-Oxygenated Components in the Ajwain Herb Oil as

Affected by Irrigation, Putrescine and Salicylic Acids and their

Interactions during Vegetative Stage (Combined Of Seasons, 2011-2012)

7 5 3 7 5 3

Control 0.97 1.35 1.84 1.39 83.09 63.46 59.30 68.62

P1 1.97 1.06 2.94 1.99 62.27 82.43 80.76 75.15

P2 1.54 1.65 3.14 2.11 82.29 63.94 62.79 69.67

S1 1.58 1.61 2.79 1.99 79.29 61.02 81.86 74.06

S2 1.70 1.15 2.03 1.63 62.13 83.03 59.87 68.34

P1S1 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

MeanIrrigation Times Irrigation Times

Oxygenated compounds

Treatment

None-oxgygenated compounds

Mean

Concerning the oxygenated and none-oxygenated compounds, the mean values of the irrigation treatmentsindicated 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-oxygenated 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 s tress revealed that P1, P1S2 and P2S1 recorded the highes t 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 simultaneously from shikimic acid or mevalonic acid. Also, the

author added that stress condition may alter the biosynthetic process to the advantage of methyl chavicol from shikimic

acid / or meva lonic acid, s ince green ruffles basil varieties are dual biosynthetic pathways.

Yield

Table 8: Herb Fresh (ton/fed) and Dry Yield (ton/fed) of Ajwain Plant as Affected by Irrigation, Putrescine,

Salicylic Acids and their Interactions, during Vegetative Stage (Combined of Seasons 2011-2012)

Character

7 5 3 7 5 3

Control 6.55 1.99 1.01 3.18 0.79 0.26 0.14 0.39

P1 3.54 4.14 0.87 2.85 0.44 0.55 0.12 0.37

P2 2.99 1.39 0.84 1.74 0.38 0.19 0.12 0.23

S1 5.40 2.37 1.07 2.95 0.65 0.31 0.15 0.37

S2 5.58 2.00 0.88 2.82 0.69 0.27 0.13 0.36

P1S1 2.70 1.79 0.74 1.74 0.33 0.24 0.11 0.23

P1S2 3.77 2.45 0.85 2.36 0.46 0.32 0.12 0.30

P2S1 2.76 1.23 0.88 1.62 0.35 0.17 0.13 0.21

P2S2 3.29 1.78 0.94 2.01 0.41 0.24 0.14 0.26

Mean A 4.07 2.13 0.90 0.50 0.28 0.13

LSD A=0.62 B=1.07 AB=1.86 A=0.04 B=0.09 AB=0.16

Herb Fresh Yield (ton/Feddan) Herb Dry Yield (ton/Feddan)

TreatmentIrrigation Times Irrigation Times

Mean (B) Mean (B)





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 s tage.

Regarding the effect o f salicylic acid and Putrescine on yield of Ajwain plant, results revealed that control without

both putrescine and salicylic ac id recorded the highest significant values of herb fresh and dry yield. Whereas application

of putrescine once at concentration of 1 ppm (P1) and salicylic acid at concentration of 100 ppm (S1) and 150 ppm (S2)

ppm as well as the combination (P1S2) were recorde d 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 fruit 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 fruit could e xplain the trend toward increase in fruit size from 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 ppm) and P1 (1 ppm 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 ppm) and irrigated every 20

days.

CONCLUSIONS

Drought s tress s ignificantly 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 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 treatments. The oil % and oil content

per plant as well as oil yield per Feddan significantly increas ed by S1 and S2 in those irrigated every 40 days. It was found

α – 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.

REFERENCES

1. Abdel Aziz, N.G.; Taha, L.S. and Ibrahim, S.M.M. (2009) Some studies on the effect of putrescine, ascorbic Acid

and thiamine on growth, flowering and some chemical constituents of gladiolus plants at Nubaria. Ozean J. of

Appl. Sci., 2(2):169 – 179.

2. Abdollahi, M.; Jafarpour, M. and Zeinali, H. (2011) Effect of var ious s alicylic ac id concentrations on growth of

Aloe vera L. International Journal of AgriScience, 1(5):311 – 313.





3. Aftab, T.; Khan, M.M.A.; Idrees, M.; Naeem, M. and Moinuddin(2010). Salicylic acid acts as potent enhancer of

growth, photosynthesis and artemisinin production in Artemisia annua L. J. Crop Sci. Biotech, 13(3):183 – 188.

4. Akhtar, J.; Ahmad, R.; As hraf, M.Y.; Tanveer, A.; Waraich, E.A. and Oraby, H.(2013). Influence of exogenous

application of Salicylic acid on s alt-stressed Mungbean (Vigna radiata): growth and nitrogen metabolism. Pak. J .

Bot., 45(1):119 – 125.

5. Alavi-samani, S.M.; Pirbalouti, A.G.; Kachouei, M.A. and Hamedi, B.(2013). The influence of reduced irrigation

on herbage, essential oil yield ad quality of Thymus vulgaris and Thymus daenensis. Journal of Herbal Drugs,

4(3):109 – 113.

6. Aly, A.A. (1987). Physiological s tudies on increasng the tolerance of pepper and broad bean plants to saline water

irrigation. Ph.D. Thes is, Agric. Bot. Dep. Fac. Agric., Cairo Univ., Egypt.

7. Azhar, N.; Hussain, B.; Ashraf, M.Y. and Abbasi, K.Y.A.R.(2011). Water stress mediated changes in growth,

physiology and secondary metabolities of Desi Ajwain (Trachys permum Ammi L.). Pak. J. Bot., 43:15 – 19.

8. Bahreininejad, B.; Razmjoo, J. and Mirza, M.(2013). Influence of water st ress on morpho -physiological and

phytochemical traits in Thymus daenens is. International Journal of Plant Production, 7:155 – 166.

9. Bernstein, L. (1961). Osmotic ad justment of plants to saline media.I.Steady state.Amer. J. Bot., 48:909 – 918.

10. Boskabady, M.I.I. and Shaikhi, J. (2000). Inhibitory effect of Carum copticum on histamine (HI) receptors of

isolated guinea-pig tracheal chains. J. Ethnopharm, 69:217 – 227.

11. Bouchereau, A.; Aziz, A.; Larher, F. and Martin-Tanguy, J.(1999). Polyamines and environmental challenges:

recent development. Plant Sci., 140:103 – 125.

12. Carpita, N.C.; Sing, N.K.; Bressan, R.A.; Haseguawa, P.M.; Reuveni, M.; Binzel, M.; La Rose, P.C.; Nelson, D.

and Schnapp, S.R.(1990). Cellular mechanisms of salt and water stress tolerance in plant. Acta Hort., 280:341 –

352.

13. Charles, D.J.; Joly, R.J. and Simon, J.E.(1990). Effects of osmotic stress on the essential oil content and

compos ition of peppermint. Phytochemis try, 29:2837 – 2840.

14. Charles, D.J.; Simon, J.E.; Shock, C.C.; Feibert, E.B.G. and Smith, R.M.(1993). Proceedings of the second

national symposium: New crops, exploration, research and commercialization. In J. Janick & J. E. Simon, eds.

Effect of water stress and post-harvest handling on artemisinin content in the leaves of Artemisia annua L. John

Wiley and Sons Inc.,New York, 640 – 643 p. .

15. Chen, Y.; Guo, Q.; Liu, L.; Liao, L. and Zhu, Z. (2011). Influence of fertilization and drought stress on the growth

and production of secondary metabolites in Prunella vulgaris L. J . Med. Plants. Res., 5:1749 – 1755.

16. Chen, Z.; Silva, S. and Klessig, D. (1993). Active oxygen species in the induction of plant systemic acquired

resistance by salicylic acid. Science, 262:1883 – 1886.

17. Cragg, G.M. and Newman, D. (2005). Biodiversity: A continuing source of novel drug leads. Pure Appl. Chem,

77:7 – 24.





18. Crisosto, C.H.; Sugar, D. and Lombard, P.B. (1988). Eff f pr pry “ m ” pr

yield co mponents. Advances in Horticultural Science, 2(2):27 – 29.

19. Das, S.K.; Molar, N.K. and Mehrotra, C.L. (1990). Salt tolerance of some agricutural crops during early growth

stage. Indian J. Agric. Sci., 41(10):882 – 888.

20. Dubois, M.; Smith, F.; Gilles, K.A.; Hamilton, J.K. and Rebers, P.A.(1956). Colorimetric method for

determination of sugars and related substances. Anal.Chem., 83(3):703 – 708.

21. El-Bassiouny, H.M.S. and Bekheta, M.A. (2005). Effect of salt s tress on relative water content, lip id peroxidation,

polyamines , amino acids and ethylene of two wheat cultivars. Int. J. Agric. Biol., 7(3):363 – 368.

22. El-Ghorab, A.H.; Mahgoub, M.H. and Bekheta, M. (2006). Effect of some bioregulators on the chemical

composition of essential oil and its antioxidant activity of Egyptian Carnation (Dian thus caryophyllus L.). Jeobq,

9(3):214 – 222.

23. El-Tarawy, M.A.; El-Mahrouk, E.M.; Ahmed, S.K. and Shala, A.Y.E.(2012). Response of Fennel plants to

chemical fertilization and Ascorbic and Saliclic acids treatments. J. Agric. Res. Kafer El-Sheikh Univ, 38(3):401 –

419.

24. EL-Tayeb, M.A. (2005). Response of barley grains to the interactive effect of salinity and salicylic acid. Plant

Growth Regul., 45:215 – 224.

25. El-Tohamy, W.A.; El-Abagy, H.M. and EL-Greadly, N.H.M. (2008). Studies on the effect of Putrescine , Yeast

and Vitamin C on growth , y ield and phys iological responses of eggplant (Solanum melongena L .) under sandy

soil conditions. Australian Journal of Basic and Applied Sciences, 2(2):296 – 300.

26. Fariduddin, Q.; Hayat, S. and Ahmad, A. (2003). Salicylic acid influences net photosynthetic rate, carboxylation,

efficiency nitrate reductase activity and seed yield in Brassica juncea. Photosynthetica, 41:281 – 284.

27. Galston, A. and Sawhney, R. (1990). Polyamines in plant phys iology. Plant Physiol., 94:406 – 410.

28. Gharib, F. and EL-Lateef, A. (2006). Effect of salicylic acid on the growth, metabolic activities and oil content of

Basil and Marjoram. International Journal of Agriculture & Biology, 8(4):485 – 492.

29. Ghasemzadeh, A. and Jaafar, H.Z.E. (2012). Effect of salicylic acid application on biochemical changes in ginger

(Zingiber officinale Roscoe). Journal of Medicinal Plants Research, 6(5):790 – 795.

30. Ghasemzadeh, A. and Jaafar, H.Z.E. (2013). Interactive e ffect of salicylic acid on some phys iological features and

antioxidant enzymes activity in ginger (Zingiber officinale Roscoe). Molecules (Basel, Switzerland), 18(5):5965 –

5979. Available at: http://www.ncbi.nlm.nih.gov/pubmed/23698049 [Accessed December 1, 2013].

31. Greenway, H. and Munns, R. (1980). Mechanisms of salt tolerance innonhalophytes. Annual Rev. Plant Physiol.,

31:149 – 190.

32. Hanafy, A.H.(1996). Physiological studies on tipburn and nutrate accumulation in lettuce plants. J. Agric. Sci.

Mansoura Univ., 21(11):3971 – 3994.

33. Hanafy Ahmed, A.H.(1997). Affect of foliar application os some chemicals on sex expression of squash plants. J.





Agric. Sci. Mansoura Univ., 22(3):697 – 717.

34. Hanafy Ahmed, A.H.(1991). Phys iological studies on the nitrogen and phosphorus deficiencies in spanish plants

(Spinacia oleracea, L.). I. Growth and organic components. Bull.Fac.Agric.Cairo Univ., 42(1):87 – 110.

35. Hanafy Ahmed, A.H.; Gad, M.M.A.; Hassan, H.M. and Amin, M.A.(2002). Improving growth and chemical

compos ition of Myrtus communis grown under soil salinity conditions by polyamines foliar applicat ion. In Minia

1st Conf. for Agric. & Environ. Sci. Mini.

36. Hayat, S.; Fariduddin, Q.; Ali, B. and Ahmad, A.(2005). Effect of salicylic acid on growth and enzyme activities

of wheat seedlings . Acta Agron. Hung., 53:433 – 437.

37. Hussein, M.M.; Balbaa, L.K. and Gaballah, M.S. (2007). Salicylic acid and salinity effects on growth of Maize

plants . Research Journal of Agricu lture and Biological Sciences, 3(4):321 – 328.

38.

Hussein, M.M.; El-Gereadly, N.H.M. and El-Desuki, M. (2006). Role of puterscine in resistance to salinity of pea plants (Pisum sativum L.). J. of Appl. Sci. Research, 2(9):598 – 604.

39. Ibrahim, M. and Jaafar, H.(2011). Involvement of carbohydrate, protein and phenylanine ammonia lyase in up -

regulation of secondary metabolites in Labisia pumila under various CO2 and N2 levels. Molecules, 16:4172 –

4190.

40. Jaleel, C.A.; Sankar, B.; Murali, P..; Gomathinayagam, M. and Lakshmanan, G.M.A Panneerselvam, R.(2008).

Water deficit stress effects on reactive oxygen metabolism in Catharanthus roseus; impacts on ajmalicine

accumulation. Colloids Surf. B: Biointerfaces, 62:105 – 111.

41.

Kakkar, R.K.; Nagar, P.K.; Ahuja, P.S. and Rai, V. k. (2000). Polyamines and plant morphogenesis. Boil.Plant.,

43:1 – 11.

42. Kang, H. and Saltveit, M.E. (2002). Chilling tolerance of maize, cucumber and rice seedling leaves and roots are

differentially affected by salicylic acid. Phys iologia Plantarum, 115:571 – 576.

43. Kang, S.; Jung, H.; Kang, Y.; Yun, D.; Bahk, J.; Yang, J. and Choi, M.(2004). Effects of methyl jasmonate and

salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root

cultures of Scopolia parviflora. Plant Sci, 166:745 – 751.

44. Khan, M.A.; Ungar, I.A. and Showalter, A.M. (2000). The effect of salinity on growth, water status, and ion

content of leaf of succulent perennial halophytes Suciea fruticosa L. J. Arid Environ., 45:73 – 84.

45. Khodary, S.E.A. (2004). Effect of salicylic acid on the growth, photosynthesis and carbohydrate metabolism in

salt stressed maize plants. J. Agric. Biol., 6:5 – 8.

46. Krishnamurthy, R. (1991). Amelioration of salinity effect in salt tolerant rice (Oryza sativa L.) by foliar

application of putrescine. Plant Cell Physiol., 32(5):699 – 703.

47. Lamb, C. and Dixon, R. (1997). The oxidative burst in plant disease resistance. Annu Rev Plant Biol, 48: 251 – 275.

48. Lawrence, B.I.M.(1988). A future examination of the variation of Ocimum basilicum L. in flavors and fragrances.

In B. M. Lawerence, B. D. Mookherjee, & B. j. W illis, Eds. A world perspective. El sevier, New york.





49. Lisar, S.Y.S.; Motafakkerazad, R.; Hossain, M.M. and Rahman, I.M.M.(2012). Introductory chapter water stress

p : causes, effects and responses.

50. Ma, J.Y.; Zhou, R. and Cheng, B.S.(1996). Effect of spermine on the peroxidase activity of detached wheat

leaves. J. Shandang Agric.Univ., 27:176 – 180.

51. Manivannan, P.; Jaleel, C.A.; Sankar, B.; Kishorekumar, A.; Somasundaram, R.; Panneerselvam, G.M. and Alagu

Lakshmanan, R.(2007). Growth, biochemical modifications and proline metabolism in Helianthus annuus L. as

induced by drought stress. Colloids Surf. B: Biointerfaces, 59:141 – 149.

52. Matthews, R.B.; Azam-Alisn and Peacock, J.M.(1990). Respons e of four sorghum lines to mid-season drought: II.

Leaf characteristics. Filed Crops Res., 25:297 – 308.

53. Mohamed, M.A. and Abdu, M.(2004). Growth and oil production of fennel (Foeniculum vulgare Mill.), effect of

irrigation and organic fertilizat ion. Bio. Agric. and Horti., 22:31 – 39.

54. Moharekar, S.T.; Lkohande, S.D.; Hara, T.; Tanaka, A. and Havan, P.D.(2003). Effect of salicylic acid on

chlorophyll and carotenoid contents of wheat and moong seedlings. Photosynthetica, 41:315 – 317.

55. Neseim, M.R.; Amin, A.Y. and El-Mohammady, M.M.S.(2014). Effect of potass ium applied with foliar spray of

yeast on sugar beet growth and yield under drought stress. Global Advanced Research Journal of Agriculture

Science, 3(8):211 – 222.

56. Nicholson, RL Hammerschmidt, R.(1992). Phenolic compounds and their role in diseas e resistance. Ann. Rev.

Phytol, 30:369 – 371.

57.

Noreen, S.; Ashraf, M. and Akram, N.A. (2011). Does exogenous application of salicylic acid improve growth

and some key physiological attributes in sunfl ower plants subjected to salt stress? Journal of Applied Botany and

Food Quality, 84:169 – 177.

58. Nornai, R. (1982). Formula for determination of chlorophyllous pigments extracted with N.N.dimethyl

formamide. Plant Physiology, 69:1371 – 1381.

59. Sangwan, N.; Farooqi, A.; Shabih, F. and Sangwan, R.(2001). Regulation of essential oil production in plants.

Plant Growth Regulation, 34:3 – 21.

60. Shakirova, F.M.; Sakhabutdinova, A.R.; Bezrukova, M.V.; Fathutdinova, R.A. and Fathutdinova, D.R.(2003).

Changes in hormonal status of wheat seedlings induced by salicylic acid and salin ity. Plant Sci., 164:317 – 322.

61. Sharafzadeh, S. and Zare, M. (2011). Influence of growth regulators on growth and secondary metabolites of

some medicinal plants from lamiaceae family. Advances in Enviromental Biology, 5(8):2296 – 2302.

62. Shi, Q.; Bao, Z.; Zhu, Z.; Ying, Q. and Qian, Q.(2006). Effects of different treatments of salicylic acid on heat

tolerance, chlorophyll fluorescence, and antioxidant enzyme activity in seedlings of Cucumis sativa L. Plant

Growth Regul, 48:127 – 135.

63. Sirvent, T. and Gibson, D.(2002). Induction of hypericins and hyperforin in Hypericum perforatum L. in response

to biotic and chemical elicitors. Physiol Mol Plant Pathol, 60:311 – 320.





64. Snedecor, G. and Cochran, W. (1980). Statistical Methods. 7 th. . USA: Press Amer, Iowa, ed. Iowa Stat Univ.

65. Staniszewska, I.; Krolicka, A.; Malinski, E.; Lojkowska, E. and Szafranek, J. (2003). Elicitation of secondary

metabolites in in v itro cultures of Ammi majus L. Enzymes Microb. Technol., 33:565 – 568.

66. Swain, T. and Hillis, W.(1959). The quantitative analysis of phenolic constituent. Sci. Food Agric., 10:63 – 69.

67. Taiz, L. and Zeiger, E.(1991). Plant Physiology. The Benjamin Cummings Publishing Co. Inc. Calif.

68. Talaat, I.M. and Balbaa, L.K. (2010). Physiological response of sweet basil plants (Ocimum basilicum L.) to

putrescine and trans-cinnamic acid. American-Eurasian J. Agric. and Environ. Sci., 8(4):438 – 445.

69. Terzi, R.; Saglam, A.; Kutlu, N.; Nar, H. and Kadiglu, A. (2010). Impact of soil drought stress on potochemical

efeciency of photosystem II and antioxidant enzyme activities of Phseolus vulgaris cultivars. Turk J. Bot., 34:1 –

10.

70.

Vahidipour, T.H.; Vahidipour, H.R.; Baradaran, R. and Jawad Seqhatoleslami, M. (2013). Effect of irrigation and

nitrogen fertilizer on grain yield and essential oil percentage of medicinal plant Ajowan. International Journal of

Agronomy and Plant Production, 4(5):1013 – 1022

effect of putrescine and salicylic acid on ajwain plant (trachyspermum amm.) at vegetative stage...

Documents