el-saadony and h.g. zyadacurresweb.com/mejas/mejas/2017/633-650.pdf · 2017. 10. 5. · 2015/2016...

Middle East Journal of Applied Sciences ISSN 2077-4613

Volume : 07 | Issue :03 |July-Sept.| 2017 Pages: 633-650

Corresponding Author: El-Saadony, F.M., Agric. Bot. Dept., Fac. Agric., Zagazig Univ., Egypt. E-mail: [email protected]

633

Effect of foliar application with salicylic acid, garlic extract and proline on growth, yield and leaf anatomy of pea (Pisum sativum L.) grown under drought

stress

El-Saadony1, F.M., Dalia A.S. Nawar2 and H.G. Zyada2 1Agric. Bot. Dept., Fac. Agric., Zagazig Univ., Egypt. 2Horticulture Dept., Fac. Agric., Zagazig Univ., Egypt.

Received: 22 June 2017 / Accepted: 28 August 2017 / Publication date: 30 Sept. 2017

ABSTRACT Field experiment was designed in two successive seasons of 2014/2015 and 2015/2016 at the

Experimental Farm El-Khatara, Fac. Agric. Zagazig University, Egypt. Plants were foliar sprayed with salicylic acid, garlic extract, proline and tap water (control) under drought stress (100, 60 and 30% of field capacity) to examine its effects on growth, yield and leaf anatomy of pea under sandy soil conditions. Results showed that most studied traits; i.e., plant height, leaves per plant, leaf area, plant organs fresh weight, photothynthetic pigments (chlorophyll a, chlorophyll b, chlorophyll a+b and carotenoids) and yield expressed as pods per plant, seeds per pod, seeds per plant and 100-seeds weight, carbohydrates as well as leaf anatomical parameters were significantly decreased with decreasing level of irrigation up to the lowest one in 1st and 2nd seasons. On the other hand, proline content in leaves increased with decreasing level of irrigation up to the lowest one 1st and 2nd seasons. Spraying plants with salicylic acid at (100ppm), garlic extract at (5%) and proline at (100ppm) had a positive significant effect in most studied traits. In general, the most favorable treatments were foliar spray pea plants with salicylic acid at (100ppm) followed by garlic extract at (5%) then proline at (100ppm), respectively compared to control.

Key words: Pea, drought stress, salicylic acid, garlic extract, proline, growth, yield, chemical contents,

leaf anatomy.

Introduction

Pea (Pisum sativum L.) belong to Fabaceae family plants and considered as popular vegetable crop and important legume crops in Egypt for local consumption and exportation. Pea is essential source of protein in human diets due to its high content of protein, ascorbic acid, carbohydrates, balanced amino acids composition and good digestibility. In general, this crop gives high yield and ensures high profits, especially when cultivated for green pods. Therefore, it occupies a prominent position among other legumes in the Egyptian agriculture. The pea seeds are rich in protein, vitamins, complex carbohydrates, antioxidant compounds, minerals and dietary fibers (Urbano et al., 2003).

Drought effect on plant growth, membrane safety, pigments, osmotic adjustment water relations, and photosynthetic activity (Praba et al., 2009). Decreasing in yield may be through drought during the vegetative stage, through drought during reproductive development or across terminal stress at the end of the crop life (Serraj et al., 2004). Chlorophyll is decrease under water stress is mainly through damage of chloroplasts (Smirnoff, 1995). However water stress cause increase of proline content in pea (Alexieva et al., 2001). Under severe water stress, cell elongation may be slow down due to breakdown of water surge from the xylem to the surrounding extending cells (Nonami, 1998). Weakness mitosis, cell extending result in depressions of crop growth under drought (Kaya et al., 2006 and Hussain et al., 2008).

Such decline in crop growth due to drought may be across either decrease in cell elongation, cell turgor and cell volume (Banon et al., 2006), and/or due to covering of xylem and phloem vessels thus obstruction any translocations (Lavisolo and Schuber, 1998).

Garlic (Allium sativum) extract contains enzymes and more than 200 chemical compounds. Its higher contents of volatile and sulphur compounds put it in the top (Pons, 2003).

Salicylic acid (SA) is a plant growth regulator known as an endogenous marking molecule, which is implicated in different physiological processes, like growth regulation, photosynthesis, stomatal behavior , nutrient uptake and mechanisms of tolerance to abiotic stresses (Hayat et al., 2010).

Middle East J. Appl. Sci., 7(3): 633-650, 2017 ISSN 2077-4613

634

Proline is the essential amino acids that accumulate in different tissues of the plant, especially in the leaves through the effect of water stress, and that the accumulation of it has a function in the regulation of osmosis in the cell as the proline is increased in the cytoplasm to counterbalance effort osmosis cell sap (Meister, 2012). Proline is an indicator of water stress tolerance and its increase in the leaf proof that the plant suffered drought stress, also is the way the plant tolerance to drought stress.

Therefore, the aimed of this present work was to study the effects of water stress, spraying with salicylic acid, garlic extract and proline on plant morphology, plant physiology, yield and its components as well as leaf blade anatomical structure of pea plants.

Materials and Methods

Field experiment was designed in two successive seasons of 2014/2015 and 2015/2016 at El-Khatara Experimental Farm, Sharkia Governorate, Fac. Agric. Zagazig University, Egypt to study the effects of water stress, foliar spray with salicylic acid, proline and garlic extract on morphological, anatomical and yield characters and seed quality as well as anatomical structure of leaf blade of pea plants. Seeds of pea cv. Master B were obtained from Hort. Res. Inst., Agric. Res. Center, Ministry of Agric., Giza, Egypt.

Field Work Procedure:

Seeds of pea were sown on 20th October in both seasons. The experimental soil was sandy in texture and chemical properties were: organic matter 0.09 and 0.08%; available N (ppm) 7.8 and 8.3; available P (ppm) 6.04 and 4.5; available K (ppm) 11 and 12; pH (1:2.5) 7.0 and 7.4 and E.C. (1:5) 2.43 and 2.28 dSm-1 in the first and second season, respectively. The recommended NPK fertilization rate for pea plants was applied in this experiment. Garlic extract (5%):

Newly produced garlic cloves were brought. 250 g of these cloves were put in a glass beaker contains 250 ml of tap water. The beaker was put in a freezer for one day, after which, frozen beaker was left to thaw. Freezing and thawing were repeated three. Water was added to a final volume of 1 liter before filtering. Final size of the filtrate was adjusted to 1: l, before being used.

Treatments:

The experiments included 12 treatments which were all possible combinations of drought stress and some substances application. Three water regimes, i.e. 100, 60 and 30% of field capacity were used. The drought stress treatments were applied after first irrigation until plant maturity. For the natural bio-stimulants, the plants were sprayed with salicylic acid (SA) at 100ppm, garlic extract (GE) at 5% and proline at 100ppm as well as control (sprayed with tap water). All foliar treatments were applied three times at 25, 40, 55 days after sowing. The experiment was laid in a split plot design with three replicates. Water regimes were randomly distributed in the main plot and foliar spray treatments were randomly arranged in the sub plots. Nitrogen fertilizer at a rate of 30 kg N/fed. was added as ammonium sulfate (20.6 % N) in two equal doses after 30 and 45 days after sowing. Phosphorus fertilizer, as calcium superphosphate (15.5% P2 O5) at the rate of 15 kg P2O5/fed., and potassium sulfate (48 % K2O) at a rate of 24 kg K2O/ fed were applied during seed bed preparation.

Recording of Data:

The present investigation involved studies pertaining to morphological, anatomical, yield and seed quality characters of pea as affected by drought stress, salicylic acid, garlic extract and proline in both studied seasons.


635

Morphological traits of vegetative growth:

A random sample of 12 plants for each treatment (3 plants from each replicate) was assigned for investigation. Vegetative characters were recorded after 60 days from sowing; i.e., 5 days after the third application of treatments. The following traits were studied in both growing seasons: Plant height, number of both branches and leaves per plant, roots fresh weight, stem fresh weight, leaves fresh weight, shoot fresh weight (stem + leaves) and leaf area per plant. Leaf area was measured by the disk method (Watson and Watson, 1953).

Photosynthetic pigments:

Chlorophyll was determined according to Saric et al. (1967). Total chlorophyll contents, chlorophyll a and chlorophyll b were calculated by use of the equations of Arnon (1949).

Chl a (mg/g-1) = [(12.7 × A663) - (2.6 × A645)] × ml acetone/mg leaf tissue (ii) Chl b (mg/g-1) = [(22.9 × A645) - (4.68 × A663)] × ml acetone/mg leaf tissue (iii) Total Chl = Chl a + Chl b.

Proline content:

The concentration of proline was expressed as mmol per gram of fresh weight (Bates et al., 1973).

Yield and its components:

Mature green pods were continuously harvested at suitable maturity stages and the following data were calculated in each of the two growing seasons: pods per plant, seeds per pod, seeds per plant, seeds weight/plant and 100-seeds weight. Chemical analysis of seeds (Seed quality):

- Percentages of nitrogen, phosphorus, potassium, carbohydrates and crude protein were determined in mature dried seeds, at harvest time of pea plants affected by drought stress, foliar spray with salicylic acid, garlic extract and proline. Samples resembling various treatments beside the seeds of control plants were finely ground. - The percentage of total carbohydrates in the seeds: were determined as glucose after acid hydrolysis and spectrophotometrically determined (Dubois et al., 1956). - The percentage of N, P, K and protein in pea seeds:

Percentage of potassium was determined by flame-photometer method (Schwarzenbach and Biedermann, 1948) and phosphorus calorimetrically (Woods and Mellon, 1985). Nitrogen content of seeds was multiplied by 6.25 to calculate the crude protein (Anon, 1990). Anatomical studies:

It was prepared to carry out a comparative microscopical testing on plant material which showed the most positive response of plant growth to tested treatments with control. Examined materials included lamina of pea leaf. Samples were taken throughout the second growing season of 2015/2016 at the age of 60 days from sowing date. Specimens were killed and fixed for at least 48 hours in F.A.A. (10ml formalin, 5ml glacial acetic acid and 85ml ethyl alcohol 70%). The selected materials were washed in 50% ethyl alcohol, dehydrated in a normal butyl alcohol series, embedded in paraffin wax of melting point 56°C, sectioned to a thickness of 20 micro-meter double stained with crystal violeterythrosin, cleared in xylene and mounted in Canada balsam (Nassar and El-Sahhar, 1998). Sections were read to detect histological manifestations of the noticeable responses resulted from tested treatments and photomicrographed.


636

Statistical analysis:

All recorded data were subjected to the analysis of variance procedures and treatment means were compared using the L.S.D. at 0.05 as described by Gomez and Gomez, 1984. The statistical analysis was done by using the computer program M-STATC software version 4.

Results and Discussion

Plant growth: Effect of drought stress:

Data in Table (1) show that plant height, number of branches and leaves/plant, Leaf area, roots fresh weight, stems fresh weight, leaves fresh weight and shoot fresh weight were significantly decreased with decreasing level of irrigation up to the lowest one in 1st and 2nd seasons except number of branches per plant in two seasons.

These mean that irrigation pea plants grown under sandy soil conditions at 100% of field capacity (FC) increased morphological characters. Plants grown under drought condition have a lower stomatal conductance. Consequently, CO2 fixation is reduced and photosynthetic rate decreases. These results were in agreement with those obtained by (Pasin et al., 1991) on grown bean.

Effect of foliar spray with SA, GE, and proline:

Data in Table 1 show that foliar spray pea plants with salicylic acid at 100ppm, garlic extract at 5% and Proline at 100ppm alone significantly increased most above-mention traits in the 1st and 2nd seasons compared to control (spraying with tap water) except roots fresh weight in 2nd season. Spraying pea plants with salicylic acid at 100ppm gave the tallest plants and recorded the maximum values of above-mention traits in both seasons. Similar results were obtained by Noor El-Deen (2005) on Majorana hortensis. Also, Gamal El-Din and Abd El-Wahed (2005) showed that a foliar application of 100 mgL1 proline increased plant height, number of branches, fresh and dry weights of (Matricaria chamomilla L. Rausch) SA application to corn and soybean promoted leaf area of plants (Khan et al., 2003). Effect of interaction between water stress and spraying with SA, GE and proline:

In general, data in Table 2 show that the interaction between irrigation at 100% of field capacity and foliar spray with SA or with GE increased plant height, Leaf area, roots fresh weight, stems fresh weight, leaves fresh weight and shoot fresh weight in two seasons of study. The interaction treatment of salicylic acid (100ppm) × control water stress (30%) and control treatment (spray with tap water) × water stress (60%) recorded the maximum and minimum values of above-mention traits, respectively.

Photosynthetic pigments:

Regarding the effect drought stress on photosynthetic pigments, data in Table (3) show that chlorophyll chl. a, chl. b, chl. a+b and carotenoids significantly decreased with decreasing level of irrigation up to the lowest one in two seasons. These results were in the same line with those showed by Abass and Mohamed (2011) on common bean, El-Ghinbihi and Hassan (2007) on pepper, Kiani et al. (2008) on sunflower (varieties), Kirnak et al. (2001) on eggplants.

The decrease in the photosynthetic activity under drought stress may be through stomatal or non-stomatal mechanisms. Stomata closure is one of the first responses to drought stress which result in declined rate of photosynthesis. The drought induced reduction in the chlorophyll content could be attributed to loss of chloroplast membranes, excessive swelling and the appearance of lipid droplets.

Table 1: Effect of drought stress and foliar spray with SA, GE and proline on plant growth of pea plants at 60 days after sowing during 2014/2015 and 2015/2016 seasons.

Main effects

Plant height (cm)

No. of branches/plant

No. of leaves/plant

Leaf area (cm2)

Roots fresh weight (g)

Stem fresh weight (g)

Leaves fresh weight (g)

Shoot fresh weight (g)

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

Drought stress

100% FC 38.75a 40.83a 3.00 3.08 30.33a 32.25a 36.35a 38.06a 2.88a 2.94 a 5.24a 6.27a 11.06a 13.16a 16.31a 19.43a 60% FC 34.58b 37.25b 2.58 2.83 29.00a 31.17a 30.41b 31.29b 2.72b 2.81b 4.96b 5.89b 8.60b 10.29b 13.56b 16.18b 30% FC 30.08c 32.50c 2.42 2.58 21.42b 23.50b 25.18c 26.17c 2.45c 2.73c 4.61c 4.51c 6.38c 7.60c 10.99c 12.11c LSD0.05 1.99 4.49 0.65 1.32 2.59 3.69 2.52 1.74 0.021 0.062 0.81 0.23 0.47 1.33 0.52 1.42

Treatments

Without 29.67d 32.22c 2.22b 2.00b 23.89d 26.22b 28.32c 29.49c 2.60c 2.88 4.80c 5.45b 8.04d 9.74b 12.85d 15.20b SA at

100ppm 40.22a 42.44a 3.44a 3.89a 29.56a 31.33a 33.30a 34.53a 2.78a 2.86 5.04a 5.78a 9.51a 11.31a 14.55a 17.08a

GE at 5% 35.56b 38.22b 2.67b 2.67b 28.22b 30.22a 31.01b 32.65ab 2.70b 2.79 5.01a 5.52ab 8.81b 10.46b 13.81b 15.10b Proline at 100ppm

32.44c 34.56c 2.22b 2.78b 26.00c 28.11b 29.89bc 30.70

bc 2.65c 2.77 4.90b 5.47b 8.36c 9.89b 13.26c 15.36b

LSD0.05 1.64 2.92 0.71 0.88 1.2 1.91 1.66 2.65 0.05 0.13 0.061 0.30 0.29 0.84 0.29 0.87

FC: field capacity, SA: salicylic acid, GE: garlic extract

637


Table 2: Effect of the interaction between drought stress and spraying with SA, GE and proline on plant growth of pea plants at 60 days after sowing during 2014/2015 and 2015/2016 seasons.

Water stress

Treatments

Plant height (cm)

No. of branches/plant

No. of leaves/plant

Leaf area (cm2)

Roots fresh weight (g)

Stem fresh weight (g)

Leaves fresh weight (g)

Shoot fresh weight (g)

1st Season

2nd Season

1st Season 2nd

Season 1st

Season 2nd

Season 1st

Season 2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

1st Season

2nd Season

60% FC

Without 33.00c 35.00bc 2.33bc 2.33a

27.33 29.33 31.72c 32.71bc 2.863ab 2.94ab 5.18bc 6.19ab 10.54b 12.62ab 15.74b 18.81abc

SA at 100ppm

48.00 a

49.66a 4.00a 4.33a 33.0 34.33 40.33a 41.74a 2.93a 3.02 a 5.30a 6.47a 11.77a 13.72a 17.07a 20.19a

GE at 5% 39.67

b 41.66b 3.00abc 2.66a 32.0 33.66 38.30a 40.81a 2.866ab 2.90ab 5.25ab 6.21ab 11.48a 13.46a 16.73a 19.67ab

Proline at 100ppm

34.33 c

37.0bc 2.66bc 3.0a 29.0 31.66 35.05b 36.97ab 2.85ab 2.90ab 5.24ab 6.20ab 10.43b 12.84ab 15.67b 19.04abc

60% FC

Without 29.66

de 32.33c

2.33bc 2.00a 26.0 28.66 29.78cd 30.82cde 2.67c 2.80bc 4.84d 5.61b 7.47e 9.78cd 12.31e 15.39de

SA at 100ppm

40.33 a

42.66ab 3.33ab 3.66a 32.0 34.00 31.82bc 33.05bc 2.840b 2.85bc 5.08c 6.11ab 9.72c 11.14abc 14.80c 17.25bcd

GE at 5% 34.66

c 37.66bc 2.33bc 2.66a 30.3 32.0 30.16cd 31.21bcd 2.703c 2.82bc 5.09c 6.01ab 8.74d 10.47bc 13.80d 16.48cde

Proline at 100ppm

33.66 c

36.33bc 2.33bc 3.0a 27.6 30.0 29.88cd 30.08cdef 2.680c 2.75c 4.84d 5.81ab 8.46d 9.78cd 13.30d 15.59de

30% FC

Without 26.33

f 29.33c

2.0c 1.66a 18.3 20.66 23.44e 24.91f 2.28f 2.58d 4.38g 4.55c 6.11f 6.82e 10.50f 11.37f

SA at 100ppm

32.33 cd

35.00bc 3.0abc 3.66a 23.6 25.66 27.73d 28.79cdef 2.566d 2.75cd 4.73de 4.74c 7.03e 9.05cd 11.77e 13.80e

GE at 5% 32.33

cd 35.33bc

2.66bc 2.66a 22.33 25.0 24.83e 25.93def 2.54d 2.86bc 4.69ef 4.32c 6.20f 7.46de 10.89f 11.79f

Proline at 100ppm

29.33 e

30.33c 2.0c 2.33a 21.33 22.6 24.72e 25.06ef 2.406e 2.71cd 4.62f 4.41c 6.17f 7.04e 10.79f 11.45f

LSD 0.05 2.85 9.09 1.24 2.7 3.12 6.93 3.51 5.89 0.08 0.27 0.12 0.68 0.64 2.65 0.67 2.82

638


Table 3: Effect of drought stress and foliar spray with SA, GE and proline on photosynthetic pigments and proline content in leaves of pea plants at 60 days after sowing during 2014/2015 and 2015/2016 seasons.

Main effects Chlorophyll a (mg/gm

F.wt) Chlorophyll b (mg/gm

F.wt) Chlorophyll a + b (mg/gm

F.wt) Carotenoids (mg/gm F.wt)

Proline content (µm/mg/F. wt)

1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season

Water stress

100% FC 3.25a 3.68 a 1.99 a 2.05 a 5.24a 5.74 a 3.06a 3.22 a 11.07 c 11.44 c

60% FC 2.38b 2.83 b 1.78 b 1.88 b 4.17 b 4.71 b 2.71b 2.73 b 15.04 b 15.20 b

30% FC 2.11c 2.20c 1.64 c 1.64 c 3.75c 3.84c 2.23c 2.34 c 16.95 a 16.96 a

LSD0.05 0.064 0.03 0.012 0.065 0.032 0.096 0.1 0.033 1.62 0.53

Treatments

Without 2.44c 2.65 d 1.89 a 1.76d 4.13d 4.42d 2.48d 2.55d 14.38 b 14.72 b

SA at 100ppm 2.58b 3.17 a 1.89 a 1.94 a 4.47b 5.11a 2.89a 3.00 a 13.68bc 13.58 c

GE at 5% 2.75a 2.97 b 1.85 b 1.88 b 4.60a 4.85b 2.76 b 2.93 b 13.56c 13.68 c

Proline at 100ppm 2.56c 2.83c 1.78c 1.83 c 4.34c 4.67c 2.54 c 2.59c 15.80 a 16.16 a

LSD0.05 0.06 0.002 0.005 0.03 0.007 0.03 0.03 0.003 2.12 0.60


639



640

Effect of foliar spray with SA, GE, and proline:

Data in Table 3 show that foliar spray of pea plants grown under sandy soil conditions with SA at 100ppm, GE at 5% and Proline at 100ppm significantly increased the concentration of chl. a, chl. b, total chl. (a+b) as well as carotenoids in two seasons compared to control (spraying with tap water).

Effect of the interaction between water stress and spraying with SA, GE and proline:

Data in Table 4 show that, in general, the interaction between irrigation at 100% FC and spraying with SA at 100ppm or with GE at 5% increased the concentration of chl. a, chl. b, total chl. (a+b) as well as carotenoids. The interaction treatment of salicylic acid (100ppm) × control water stress and control treatment (spray with tap water) × water stress (30%) recorded the maximum and minimum values of above-mention traits, respectively.

The interactive effect of drought stress and the usage of natural substances resulted in significant increases in photosynthetic pigments, enzymes activity and proline concentration compared with untreated plants.

Proline content in leaves: Effect of drought stress on proline content in leaves:

It is obvious from such data in table (3) that proline content in leaves significantly increased with decreasing level of irrigation water up to the lowest one in two seasons. These results coincide with those reported by Nour (2005) on cowpea. These results coincide with those reported by (Alexieva et al., 2001) and (Verbruggen and Hermans, 2008).

Effect of foliar spray with SA, GE and proline:

Data in Table 3 show that foliar spray of pea plants grown in sandy soil with salicylic acid at 100ppm, garlic Extract at 5% and proline at 100ppm alone significantly increased proline content in leaves in two seasons compared to control (spraying with tap water).

Effect of the interaction between water stress and spraying with SA, GE and proline:

Data in Table 4 show that, in general, the interaction between irrigation at 30% of FC and spraying with proline at 100ppm or with GE at 5% significantly increased proline content in both seasons.

Yield and its components: Effect of drought stress on yield of pea plants:

It is obvious from such data in Table 5 that all above-mentioned traits were significantly decreased with decreasing level of irrigation up to the lowest one in two seasons. These results coincide with those reported by (Frederick et al., 2001) on soybean, (Pilbeam et al., 1992) on spring-sown faba bean and Nour et al. (2007) on cowpea (vegna ungiculata L.).

Effect of sprayed with salicylic acid, garlic extract and proline:

Data in Table (5) show the effect of sprayed with salicylic acid, proline and garlic extract yield

of pea plants. It is obvious from such data that pea yield expressed as pods per plant, seeds per pod, seeds per plant and 100-seeds weight were significantly increased in two seasons compared to control (spraying with tap water). Similar results were reported by Ahmed et al. (2005) on pea plants treated with garlic extract.

Table 4: Effect of the interaction between drought stress and spraying with SA, GE and proline on photosynthetic pigments and proline content in leaves of pea plants at 60 days after sowing during 2014/2015 and 2015/2016 seasons.

Water stress

Treatments

Chlorophyll a (mg/gm/ F. wt)

Chlorophyll b (mg/gm/ F. wt)

Chlorophyll a + b (mg/gm/F. wt)

Carotenoids (mg/gm/ F. wt)

Proline content (µm/mg/F.wt)

1st Season

2nd Season

1st Season 2nd

Season 1st

Season 2nd

Season 1st

Season 2nd

Season 1st Season 2nd Season

100% FC

Without 3.23c 3.42c 1.91c 1.99cde 4.96c 5.41d 2.86cd 2.97e 11.25c 12.117c

SA at 100ppm

3.36a 3.90a 2.063a 2.13a 5.423a 6.030a 3.25a 3.39b 9.86cd 10.01d

GE at 5% 3.35a 3.75b 2.066a 2.09ab 5.416a 5.84b 3.22ab 3.55a 9.45d 9.75d Proline at 100ppm

3.23b 3.663c 1.93b 2.00cd 5.16b 5.66c 2.906c 2.99d 13.73b 13.87b

60% FC

Without 2.27ef 2.48h 1.636h 1.74f 3.90gh 4.22g 2.44f 2.42h 14.81b 14.79b

SA at 100ppm

2.18fg 3.28e 1.89c 2.00bc 4.083f 5.286d 3.13b 3.22c 14.30b 14.37b

GE at 5% 2.716d 2.89f 1.80d 1.90de 4.523d 4.79e 2.76d 2.83f 14.55b 14.56b Proline at 100ppm

2.37e 2.67g 1.79e 1.87e 4.156e 4.54f 2.516e 2.45g 16.5a 17.07a

30% FC

Without 1.99h 2.063l 1.540i 1.56g 3.53j 3.62k 2.14h 2.26j 17.06a 17.24a

SA at 100ppm

2.18fg 2.32i 1.72f 1.69f 3.90g 4.016h 2.29g 2.39h 16.88a 16.34a

GE at 5% 2.17fg 2.26j 1.69g 1.66f 3.866h 3.926i 2.29g 2.40h 16.68a 16.72a Proline at 100ppm

2.09gh 2.16k 1.623h 1.63f 3.716i 3.79j 2.19h 2.32i 17.16a 17.52a

LSD 0.05 0.11 0.05 0.01 0.12 0.05 0.16 0.11 0.05 1.60 1.45


641


Table 5: Effect of drought stress, foliar spray with SA, GE and proline on yield components of pea plants during 2014/2015 and 2015/2016 seasons.

Main effects and interactions

No. of pods/ plant No. of seeds/pod No. of seeds/plant 100-seeds weight (g) Seeds yield /plant (g)

1st Season

2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season

Water stress

100% FC 17.58a 18.42a 6.583 a 7.08 a 109.75 a 112.33 a 33.66a 33.82 a 37.068a 38.109a

60% FC 13.83b 14.33b 5.167 b 5.167 b 71.00 b 70.92 b 27.88b 28.166 b 19.827b 20.035b

30% FC 10.08c 10.50c 4.33c 4.50 c 44.08 c 45.33 c 25.53b 25.34c 11.075c 11.576c

LSD0.05 1.62 2.22 1.24 0.59 13.88 9.55 2.89 2.1 2.90 3.39

Treatments

Without 12.44b 12.66b 4.89 5.89 59.89 c 61.78 c 27.38b 27.48b 17.145c 17.863c

SA at 100ppm 15.22a 15.89a 5.78 5.67 86.67 a 88.444 a 30.39a 30.34a 27.001a 27.764a

GE at 5% 14.44

ab 15.44a 5.44 5.67 79.56 ab 80.44 ab 29.49a 29.63 a 24.444ab 24.936b

Proline at 100ppm

13.22 ab

13.67b 5.33 5.11 73.67 b 74.11 b 28.84 ab 28.97 a 2.20b 2.23b

LSD0.05 2.12 1.31 1.46 1.20 12.93 9.9 1.92 1.49 3.35 2.57


642



643

Chemical analysis of seeds (Seed quality): Effect of drought stress:

Data in Table (6) show the effect of drought stress on total NPK and protein as well as total carbohydrate on seeds of pea plants. It is obvious from such data that total NPK and protein as well as total carbohydrate on seeds of pea plants significantly decreased with decreasing level of irrigation up to the lowest one in two seasons.

Effect of spray with SA, GE and proline:

Data in Table 6 show that foliar spray of pea plants grown in sandy soil with salicylic acid (100ppm), garlic Extract (5%) and Proline (100ppm) alone significantly increased values of above-mention traits in two seasons compared to control (spraying with tap water) except Potassium content in seeds in the 2nd season. Spraying pea plants with salicylic acid at 100ppm gave the highest values of above-mention traits in two seasons compared to control. These results were in the same line with those showed by Noor EL-Deen (2005) on Marjoram Plants (Majorana hortensis L.).

Effect of interaction between drought stress spraying with salicylic acid, garlic extract and proline:

Data in table 7 show significant effect on all traits under study in two seasons. The interaction

treatment of salicylic acid (100ppm) × 100% of FC and control treatment (spray with tap water) × water stress (30%) recorded the maximum and minimum values of above-mention traits, respectively.

Anatomical studies:

Presented data in table 8 show that, the interaction between FC at 100% and foliar spray with SA at 100ppm gave the highest values of palisade cell length, spongy tissue thick., lamina thick., midrib V.B. width and length and midvein thicknss. For the interaction between FC at 100% and foliar spray with SA at 100ppm over the interaction between FC at 100% and foliar spray with tap water, (control). In general, the interaction between FC at 100% and foliar spray with SA at 100ppm, GE at 5% and proline at 100ppm as well as tap water increased all leaf anatomical parameters, whereas, the interaction between FC at 30% and foliar spray with SA at 100ppm, GE at 5% and proline at 100ppm as well as tap water decreased all leaf anatomical parameters (Fig. 1).

All enhanced leaf anatomical parameters (palisade cell length, spongy tissue thickness, blade thickness, midrib vascular bundle width, midrib vascular bundle length and midvein thickness) due to irrigation of pea plants at 100% of FC, and foliar application with salicylic acid (100ppm), garlic extract (5%) and proline (100ppm) reflected on a good translocation of the observed water and nutrients into cell to be used in different metabolic process which positively affected fresh weight of leaves and shoot (Table, 2) on photosynthesis process activity (Table, 4) and accumulation of photo-assimilates, there by helping in better retention of flowers and fruits and this in turn increased yield (Table, 5).

Bundle length decreased in response to spraying 100 ppm proline Fig. 1; this decrease was accompanied by increasing in leaf thickness and numbers of xylem row and vessels, yet all the recorded values Table (8) were still higher than the control plant. In this respect (Meloni et al., 2001) revealed that in stressed plants the contributory role of osmoprotectance i.e. glycinebetaine and mainly proline. However may be proline treatment helped in increasing the osmolytes synthesis in the plant.

Table 6: Effect of drought stress and foliar spray with SA, GE and proline on N, P, K and total protein and total carbohydrate on seeds of pea plants at harvest time during 2014/2015 and 2015/2016 seasons.

Main effects and interactions

N (%) Protein (%) P (%) K (%) Total carbohydrate (%)

1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd Season

Water stress

100% FC 3.903 a 4.02 a 24.40a 25.13 a 0.36 a 0.37 a 1.75 a 1.74 a 24.96 24.84 a

60% FC 3.45 b 3.49 b 21.57 b 21.84 b 0.35 b 0.35 b 1.65 b 1.66 b 23.00 23.21 b

30% FC 3.07 c 3.07 c 19.18 c 19.21 c 0.33 c 0.34 c 1.47 c 1.46 c 21.04 21.43 c

LSD0.05 0.10 0.08 0.65 0.48 0.014 0.01 0.27 0.02 0.26 0.44

Treatments

Without 3.36 c 3.39 b 20.10 c 21.21 b 0.34 b 0.35 1.59 c 1.60 b 21.88 b 22.02 c

SA at 100ppm 3.50 ab 3.59 a 21.91 ab 22.47 a 0.35 ab 0.36 1.64 ab 1.64 a 23.16 a 23.25 b

GE at 5% 3.45 bc 3.52 ab 21.57 bc 21.99 ab 0.35 ab 0.36 1.61 bc 1.60 b 23.501a 23.46 b

Proline at 100ppm 3.58 a 3.61 a 22.39 a 22.59 a 0.35 a 0.36 1.66 a 1.65 a 23.47 a 23.91 a

LSD0.05 0.12 0.13 0.73 0.81 0.011 0.02 0.042 0.03 0.47 0.41


644


Table 7: Effect of the interaction between water stress and spraying with SA, GE and proline on N, P, K, total protein and total carbohydrate of seeds of pea plants at harvest timeduring 2014/2015 and 2015/2016 seasons.

Water stress

Treatments

N (%) Total protein (%) P (%) K (%) Total Carbohydrate

(%)

1st Season 2nd Season 1st Season 2nd Season 1st Season 2nd

Season 1st Season 2nd Season 1st Season 2nd Season

100% FC

Without 3.85ab 3.97a 24.11ab 24.84a 0.36ab 0.37ab 1.74ab 1.72a 23.57b 23.44b SA at

100ppm 3.97a 4.10a 24.81a 25.50a 0.37a 0.37a 1.75a 1.763a 25.50a 25.20a

GE at 5% 3.91a 4.08a 24.23a 25.62a 0.36abc 0.37ab 1.73abc 1.74a 25.58a 25.38a Proline at 100ppm

3.87a 3.93a 24.43a 24.56a 0.36ab 0.37a 1.76a 1.746a 25.16a 25.33a

60% FC

Without 3.31cde 3.32bcd 20.69cde 20.73bcd 0.34cde 0.35ab 1.70abcd 1.72a 21.62c 21.88cd SA at

100ppm 3.46cd 3.56bc 21.65cd 22.26bc 0.35bcd 0.35ab 1.64cd 1.66b 23.48b 23.72b

GE at 5% 3.45cd 3.49bc 21.58cd 21.86bc 0.34cde 0.35ab 1.61de 1.626b 23.50b 23.57b Proline at 100ppm

3.57bc 3.60b 22.35bc 22.52b 0.35bcd 0.35ab 1.65bcd 1.643b 23.41b 23.65b

30% FC

Without 2.90f 2.88e 18.18g 18.04e 0.313f 0.33b 1.34g 1.353f 20.45d 20.74de

SA at 100ppm

3.08ef 3.14de 19.26efg 19.64de 0.33e 0.34ab 1.52f 1.503d 20.48d 20.83de

GE at 5% 3.02ef 2.97e 18.89fg 18.48e 0.333de 0.34ab 1.48f 1.436e 21.41cd 21.41de Proline at 100ppm

3.26de 3.31cd 20.37def 20.68cd 0.34cde 0.34ab 1.55ef 1.563c 21.84c 22.74bc

LSD 0.05 0.29 0.13 0.65 1.77 0.65 0.016 0.04 0.09 1.02 1.06


645


Table 8: Effect of water stress and foliar spray with SA, GE and proline on leaflet anatomical traits of pea plants during 2015/2016 season.

Treatment

Parameters

100% FC 60% FC

Without SA at

100ppm ± % to control

GE at 5%

± % to control

Proline at

100ppm

± % to control

Without ± % to control

SA at 100ppm

± % to control

GE at 5%

± % to control

Proline at 100ppm

± % to control

Lamina thick. (µ)

320 350 + 9.30 330 + 3.12 340 + 6.25 300 - 6.25 320 - 310 - 3.10 290 - 9.37

Palisade cell length (µ)

100 110 + 15 100 - 110 + 15 95 - 5.0 100 - 110 + 15 100 -

Spongy tissue thick. (µ)

185 190 + 2.7 185 - 185 - 160 -13.50 170 - 8.1 160 - 13.5 160 - 13.5

Midvein thick. (µ)

620 750 + 20.9 650 + 4.83 620 - 500 -19.3 580 - 6.45 550 - 11.2 540 - 12.9

Midrib V.B. length. (µ)

260 330 + 26.9 320 + 23.1 320 + 23.1 250 - 3.8 260 - 270 + 3.84 260 -

Midrib V.B. Width. (µ)

150 200 + 33 200 + 33 170 + 13..3 160 - 6.6 160 + 13.3 170 + 13.3 160 + 6.6

Table 8: Cont.

Treatment

Parameters

30% FC

Without ± % to control

SA at 100ppm ± % to control

GE at 5% ± % to control

Proline at 100ppm ± % to control

Lamina thick. (µ) 270 - 15.6 285 - 10.9 280 - 12.5 280 - 12.5

Palisade cell length (µ) 90 - 10 95 - 5.0 95 - 5.0 95 - 5.0 Spongy tissue thick. (µ) 120 - 35.1 150 - 18.9 150 - 18.9 150 - 18.9 Midvein thick. (µ) 470 - 24.1 520 - 16.1 510 - 17.7 480 - 22.6

Midrib V.B. length. (µ) 220 - 15.3 235 - 9.6 230 - 11.5 230 - 11.5

Midrib V.B. Width. (µ) 150 - 170 + 13.3 150 - 120 - 20

646



647

100% of FC + spray with tap water

100% of FC + spray with salicylic acid

100% of FC + spray with garlic extract

100% of FC + spray with proline

Fig. 1: Effect of water stress treatment of pea plants, sprayed with salicylic acid, proline,

garlic extract and their interactions on leaflet anatomy at 60 days after sowing at

the season of 2015/2016.


648





Fig. 1. Cont: Effect of water stress treatment of pea plants, sprayed with salicylic acid,

proline, garlic extract and their interactions on leaflet anatomy at 60 days after

sowing at the season of 2015/2016.

References Abass, S.M., H.I. Mohamed, 2011. Alleviation of adverse effects of drought stress on common bean

(Phaseolus vulgaris L.) by exogenous application of hydrogen peroxide. Bangladesh J. Bot., 41: 75-83.

Ahmed, S., J. Iqbal and I. Attauddin, 2005. Time of application effect of phytobiocides on powdery mildew and yield in pea. Sarhad J. Agriclture, 21: 729-731.


649

Akram, H.M., A. Ali, A. Sattar, H.S.U. Rehman and A. Bibi, 2013. Impact of water deficit stress on various physiological and agronomic traits of three Basmati rice (Oryza sativa L.) cultivars. J. Anim. Plant Sci., 23(5): 1415-1423.

Alexieva, V., I. Sergiev, S. Mapelli, E. Karanov, 2001. The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ, 24: 1337-1344.

Anon., 1990. Official Methods of Analysis of the Association of the Official Analytical Chemists (A.O.A.C.). 15th Edit., Published by A.O.A.C.D.C. Washington.

Arnon, D.I., 1949. Copper enzymes in isolated chloroplasts, polyphenoxidase in beta vulgaris. plant physiology, 24: 1-15.

Banon, S.J., J. Ochoa, J.A. Franco, J.J. Alarcon and M.J. Sanchez-Blanco, 2006. Hardening of oleander seedlings by deficit irrigation and low air humidity. Environ Exp. Bot., 56: 36-43.

Bates, L., R. Waldren and I. Teare, 1973. Rapid determination of free proline for water-stress studies. Plant and soil, 39(1): 205-207.

Dubois, M., R.A. Gilles, J. Hamillon, R. Rebers and I. Smith, 1956. Colorimetric method for determination of sugars and related substances. Anal. Chem., 28: 350.

El-Ghinbihi, F.H. and M.I. Hassan, 2007. Effect of some natural extracts and ascorbic acid as foliar spray on growth, leaf water contents, chemical composition and yield of pepper plants grown under water stress conditions. Menofia J. Agri. Res., 32: 683-710.

Frederick, J.R, C.R.Camp and P.J. Bauer, 2001. Drought-stress effects on branch and main stem seed yield and yield components of determinate soybean. Crop Sci., 41: 759-763.

Gamal El-Din K.M. and M.S.A. Abd El-Wahed, 2005. Effect of some amino acids on growth and essential oil content of chamomile plant. Int. J. Agric. Biol., 7: 376-380.

Gomez, K.A. and A.A. Gomez, 1984. Statistical procedures for agricultural research. John Wiley and sons, Inc. London, UK (2nd ed) 13-175.

Hayat, Q., S. Hayat, M. Irfan and A. Ahmad, 2010. Effect of exogenous salicylic acid under changing environment: A review. Environ. Exp. Bot., 68: 14-25.

Hussain, M., M.A. Malik, M. Farooq, M.Y. Ashraf, M.A. Cheema, 2008. Improving Drought tolerance by exogenous application of glycinebetaine and salicylic acid in sunflower, J. Agron. Crop Sci., 194: 193-199.

Kaya, M.D., G. Okçub, M. Ataka, Y. Çıkılıc and Ö. Kolsarıcıa, 2006. Seed treatments to overcome salt and drought stress during germination in sunflower (Helianthus annuus L.). Eur. J. Agron., 24: 291-295.

Khan, W., B. Prithviraj and D.L. Smith, 2003. Photosynthetic responses of corn and soybean to foliar application of salicylates. J. Plant Physiol., 160: 485-492.

Kiani, R., T.D. Hanks and M.N. Shadlen, 2008. Bounded integration in parietal cortex underlies decisions even when viewing duration is dictated by the environment. Journal of Neuroscience, 28:3017-3029.

Kirnak, C., I.T. Kaya and D. Higgs. 2001. The influence of water deficit on vegetative growth, physiology, fruit yield and quality in eggplants. Bulg. J. Plant Physiol., 27:34-36.

Lavisolo, C. and A. Schuber, 1998. Effects of water stress on vessel size xylem hydraulic conductivity in (Vitis vinifera L). J. Exp. Bot., 49(321): 693-700.

Meister, A. 2012. Biochemistry of the amino acids, Elsevier. Meloni, D.A., M.A. Oliva, H.A. Ruiz and C.A. Martinez, 2001. Contribution of proline and inorganic

salutes to osmotic adjustment in cotton under salt stress. J. Plant Nutrition, 24: 599-612. Nassar, M.A. and K.F. El-Sahhar, 1998. Botanical Preparations and Microscopy (Microtechnique).

Academic Bookshop, Dokki, Giza, Egypt, 219pp. (In Arabic). Nonami, H. 1998. Plant water relations and control of cell elongation at low water potentials, J. Plant

Res., 111: 373-382. Noor EL-Deen, T.M. 2005. Physiological Studies on Marjoram Plants (Majorana hortensis, M.).

M.Sc. Thesis, Ornamental Hort. Dept., Fac. Agric. Moshtohor, Zagazig Univ., Banha, Egypt. Nour, K. A. M. 2005. Response of some cowpea (vegna ungiculata L.) cultivars to some irrigation

levels by drip system and organic manure under sandy soil condition. Ph.D. Thesis, Fac. of Agric. Mansuora Univ.


650

Nour, K.A.M., T.B. Ali, M. Fayza and A. Darwesh, 2007. Water equirement of cowpea as affected by organic fertilization rates under newly reclaimed soil conditions. Egy. J. Appl. Sci., 22(8B): 482-496.

Pasin, N.H., B.G. Filho, O.S.B. Santos and V.D.C. Mello, 1991. Performance of bean seeds derived from plants Subjected to water stress at two growth stages. Pesquisa Agropecuaria Brasileira, 26(2): 183-192.

Pilbeam, C.J., J.K. Akatse, P.D. Hebblethwaite and C.D. Wright, 1992. Yield production in two contrasting forms of spring-sown faba bean in relation to water supply. Field Crops Res., 29: 73-287.

Pons, L.I. 2003. Fotoproteccion Vegeta (II). Offarm 22: 163-164. Praba, M.L., J.E. Cairns, R.C. Babu and H.R. Lafitte, 2009. Identification of physiological traits

underlying cultivar differences in drought tolerance in rice and wheat. J. Agron. Crop Sci., 195: 30-46.

Saric, M., R. Kostroi, T. Cupina and I. Geric, 1967. Chlorophyll determination Univ. U.Noven Sadu Prakitikum is Kiziologize Bilijaka Beogard, Haucna, Anjiga.

Schwarzenbach, G., W. Biedermann, 1948. Komplexone X. Erdalkalikomplexe von o,6- Dioxyazofarbstoffen,. Helv. Chim. Acta., 31: 678-687.

Serraj, R., L. Krishnamurthy, J. Kashiwagi, J. Kumar, S. Chandra and J.H. Crouch, 2004. Variation in root traits of chickpea (Cicer arietinum L.) grown under terminal drought. Field Crops Res., 88: 115-127.

Smirnoff, N. 1995. Antioxidant systems and plant response to the environment. In: Smirnoff V (Ed.), Environment and Plant Metabolism: Flexibility and Acclimation, BIOS Scientific Publishers, Oxford, UK.

Urbano, G., P. Aranda and E. Gomez-Villalva, 2003. Nutritional evaluation of pea (Pisum sativum L.) protein diets after mild hydrothermal treatment and with and without added phytase. Journal of Agricultural and Food Chemistry, 51: 2415-2420.

Verbruggen, N. and C. Hermans, 2008. Proline accumulation in plants: a review. Amino Acids 35: 753-759.

Watson, D. J. and M. A. Watson, 1953. Studies in potatoes agronomy. 1- Effect of variety seed size and spacing on growth, development and yield. J. Agr. Sci., 66: 249-249.

Woods, J. T. and M. G. Mellon, 1985. Chlorostannous reduced molybdophosphoric blue color method, in sulphuric acid system. In Soil Chemical Analysis by Jackson, M. L. (1985). Printic-Hall International, Inc., London.

el-saadony and h.g. zyadacurresweb.com/mejas/mejas/2017/633-650.pdf · 2017. 10. 5. · 2015/2016...

Documents