THE EFFECT OF ASCORBIC ACID TREATMENT ON
WHEAT (TRITICUM AESTIVUM L.) SEEDLINGS
UNDER DROUGHT STRESS
Khadr, S.A. ; S.M. El-Hamamsy ; H.A. El-khamissi and Z.H. Saad
Biochemistry Dep., Faculty of Agric., Al-Azhar Univ., Cairo, Egypt
Key Words: Ascorbic acid, drought stress, chlorophyll, enzymes activity
ABSTRACT
Three Egyptian wheat (Shanadaweel 1, Giza168, and Masr 1)
varieties were evaluated under drought stress using different
concentrations of polyethylene glycol (PEG) (0, 1, 2, and 3 bars) and
investigate the effects of the application of ascorbic acid (0, 150, and 200
mg/L) in the alleviation of drought stress at the seedling stage. The
results revealed that drought stress caused a decrease in germination
percentage, growth parameters (shoot length and root length) and
photosynthetic pigments (chlorophyll A, chlorophyll B, and total
carotenoids) with increasing PEG concentrations in all wheat varieties.
Moreover, Catalase (CAT) activity increased in Giza168 with increasing
drought stress at 3 bar, whereas CAT activity increased in Masr 1 and a
nonsignificant increase in Shandaweel 1 at 1 bar and then decreased at 2
bar. Also, proline accumulation under drought stress in all wheat
varieties. On the other hand, pretreatment of seeds treated with ascorbic
acid (0, 150, 200 mg/l) enhanced all parameters studied under drought
stress. 200 mg/L of ASA is the most effective in mitigating the effects of
drought stress on the morphological and chemical characteristics of
wheat varieties at the seedling stage.
INTRODUCTION
Drought, one of the environmental stresses, is the most significant
factor restricting plant growth and crop productivity in the majority of
agricultural fields of the world (Tas and Tas 2007). Drought decreased
germination and seedling growth, and this is one important case to
produce crops (Gamze et al., 2005). Drought stress causes reduced
stomatal conductance resulting in decreased net photosynthetic rate.
Chlorophyll degradation due to drought stress also inhibits the
photosynthetic rate in wheat (Moaveni, 2011). In addition, it inhibits the
photochemical activities and decreases the activities of enzymes in the
Calvin Cycle in photosynthesis (Monakhova and Chernyadev 2002).
PEG as a factor causing drought stress by reducing water potential results
in reducing growth in seed germinated and stopping seedling growth so
that this effect has been observed more in the shoot than primary roots
(Zhu, 2006). Dodd and Donovan (1999) also suggested that PEG
prevents water absorption by seeds, but penetrable ions by reducing
Egypt. J. of Appl. Sci., 36 (1) 2021 30-42
potential inside cell results in water absorption and starting to germinate.
Proline accumulation under salinity/ drought stress (Kavi Kishor et al.,
2005), which protect the proteins against denaturation and act as osmotic
balancing agents (Sivakumar et al., 2000). The generation of ROS is
limited or scavenged by an antioxidant system, including antioxidant
compounds (ascorbate, salicylate, glutathione, tocopherol, etc.) and
antioxidant enzymes like catalase (CAT), superoxide dismutase (SOD)
and peroxidase (POD) (Foyer and Noctor, 2003). Catalase (CAT) and
peroxidase (POD) are enzymes that catalyze the conversion of H2O2 to
water and O2 (Gratao et al., 2005).
Wheat (Triticum aestivum L.) is one of the most important crops
in Egypt, which plays a special role in people’s nutrition. Ninety-five
percent of the cultivated wheat is used for the preparation of bread and
other baked products, while the remaining 5% is durum wheat, which is
used essentially for making pasta and macaroni (Bushuk, 1998). External
application of ascorbic acid activates antioxidant mechanisms, and this
improves resistance to drought stress (Shalata and Neumann, 2001).
Pre-sowing treatment with ascorbic acid is widely used. It improves
performance and stand establishment at different external factors such as
high salinity (Shaddad et al. 1990 and Afzal et al. 2005). The objectives
of this investigation are to study the effects of ascorbic acid applications
as an antioxidant on seeds germination, growth parameters,
Photosynthetic pigments, proline, and activity of enzymes in three wheat
seedlings grown under drought stress.
MATERIALS AND METHODS
Plant material and treatments: -
The present study was carried out in Biochemistry Department, Faculty
of Agriculture, Al-Azhar University, three varieties of wheat (Triticum
aestivum L.) included Shandaweel 1, Giza168 and Masr 1 were used in
this study. wheat seeds were obtained from the Department of Seed
Technology Research, Field Crops Research Institute, Agriculture
Research Center (ARC), Ministry of Agriculture, Giza, Egypt.
Homogeneous seeds were sterilized using 70% ethanol for 2 min.
followed by 0.2% sodium hypochlorite (NaOCl) for 3 min. then rinsed
for 3 times with distilled water.
Application of Ascorbic acid and growth conditions
After washing the seeds, they were soaked for 24 h at room
temperature in distilled water or 150 and 200 mg/L Ascorbic acid before
sowing in Petri dishes. Effect of drought stress induced by different
osmotic potential level (0, 1, 2 and 3 bars) of polyethylene glycol 8000
(PEG 8000) (Michel, 1983). Germination trials were carried out in 15 cm
Petri dishes containing a layer of two filter paper whatman’s two filter
31 Egypt. J. of Appl. Sci., 36 (1) 2021
paper sterilized with distilled water or PEG solution. Three replications
were sown in Petri dishes on two filter paper beds and each treatment
contained 50 pure seeds, then irrigated with 10 ml solution of different
concentration of PEG solution (0, 1, 2 and 3 bars) and incubated in
growth chamber at 20±2 °C for 10 days. Seed germination was observed
daily with fresh PEG solution added to the Petri dishes as necessary to
maintain moisture levels. Germination percentage was calculated using
the formula outlined by Krishnasamy and Seshu (1990). Seedling
shoots and roots lengths of ten randomly selected seedlings were
measured after 10 days of germination (ISTA, 1993).
Biochemical analysis: -
Determination of of photosynthetic pigments:-
To extract photosynthetic pigment from wheat shoots, 0.2 g of
fresh shoot was homogenized with 10 ml of 100% acetone in a porcelain
mortar and centrifuged at 2500 rpm for 10 min. The supernatant was
separated and the absorbances were read at 662 and 645 nm for
chlorophyll a and b and 470 nm for carotene. Were calculated using
following equations of Lichtentaler and Wellburn (1985).
Chlorophyll a (μg/ml) = 11.75 A662 – 2.350 A645
Chlorophyll b (μg/ml) = 18.61 A645 – 3.960 A662
Carotenoids (μg/ml) = 1000 A470 – 2.270 Chl a – 81.4 Chl b/227.
Extraction of antioxidant enzymes:-
Enzymes were extracted from 0.5 g leaf samples homogenized in
a pre-chilled pestle and mortar containing ice cold 0.1 M phosphate
buffer (pH 7.5) and 0.5 mM EDTA. Each homogenate was transferred to
centrifuge tubes and centrifuged at 4°C in a Sorval model T21 (Thermo
Scientific, Waltham, MA) refrigerated centrifuge for 15 min at 15000 x
g. The supernatant was decanted and used for measuring enzyme activity
assays (Esfandiari et al., 2007).
Enzymes activity assay:-
Catalase activity:-
Catalase activity was determined according to the method used by
Aebi (1984) in which the disappearance of H2O2 in a reaction mixture
containing 0.3 mL 3% H2O2, 2.5 mL of 0.05 M phosphate buffer (pH 7),
and 2.5 mL of plant extract is monitored by the decrease in absorbance at
240 nm.
Peroxidase activity:-
Peroxidase was assayed spectrophotochemically according to
(Amako et al., 1994) the assay was carried out at 25 °C in 1.0 cm light
Egypt. J. of Appl. Sci., 36 (1) 2021 32
path cuvette and the reaction mixture consisted of 1500 μL phosphate
buffer, 1000 μL pyrogallol and 480 μL H2O2 solution. After mixing, the
reaction was initiated by adding the enzyme extract (20 μL) and the
increase in optical density at 430 nm against blank (without extract) was
continuously recorded every minute (for 1 min).
Determination of proline:-
Proline content of shoot was determined according to a
modification of the method of Bates et al (1973). Samples of shoots (0.5
g) were homogenized in a mortar and pestle with 10 ml sulfosalicylic
acid (3% w/v), and then centrifuged at 18,000 g for 15 min. Two ml of
the supernatant was then added to a test tube, to which 2 ml glacial acetic
acid and 2 ml freshly prepared acid ninhydrin solution (1.25 g ninhydrin
dissolved in 30 ml glacial acetic acid and 20 ml 6 M phosphoric acid)
were added. The test tubes were incubated in a water bath for 1 h at
100°C and then allowed to cool to room temperature. Four ml of toluene
were then added to the tubes and then mixed on a vortex mixer for 20 s.
The test tubes were allowed to stand for at least 10 min to allow
separation of the toluene and aqueous phases. The toluene phase was
carefully pipetted out into a glass test tube and its absorbance was
measured at 520 nm in a spectrophotometer. The content of proline was
calculated from a standard curve and calculated on a fresh weight basis
as follows: {(μg proline/ml × ml toluene) / 115.5 μg/μmole}/ {(g
sample)/5} = μmoles proline/g of fresh weight material.
Statistical analysis:
Randomized Complete block design analysis for all data obtained
was carried out and differences between means were calculated using
L.S.D test according to Steel and Torrie (1980).
RESULTS AND DISCUSSION
The effect of ascorbic acid on germination and growth parameters in
three wheat varieties under drought stress by PEG
The effect of ascorbic acid on germination percentage and growth
the parameters under drought stress are presented in Table (1). Under
control, the highest values of germination percentage were observed in
Giza168 and Shandweel 1 (98%). Germination percentage decreased by
increasing of drought stress by PEG level. Germination percentage in all
wheat varieties were significantly decreased at 1 and 2 bars but Giza168
has a higher germination percentage (86%) at 2, while the lowest value of
germination percentage was found in Masr 1 variety (56%) at the same
level of drought stress. At 3 bar drought stress germination percentage
was (0%) in all wheat varieties. From the results in Table (1) indicated
that Giza168 more drought tolerant than Shandaweel 1 and Masr 1.
33 Egypt. J. of Appl. Sci., 36 (1) 2021
Table (1): The effect of ascorbic acid on germination and growth
parameters in three wheat varieties under drought
stress by PEG.
Application of ascorbic acid (ASA) as soaking led to increasing
of germination percentage in all wheat varieties (Table 1). 150 mg/L of
ASA caused significantly increase in Shandaweel 1 and Giza168 from
(76 and 86%) to (81 and 92%), respectively at 2 bar , while led to
decrease in Masr 1 from 56% to 53% at 2 bar, 150 mg/L of ASA was not
effective on germination in all wheat varieties at 3 bar. Application of
200 mg/L of ASA caused increasing of germination in all wheat
varieties. Germination percentage increased from 0% to 65, 85 and 60 %
at 3 bar in Shandaweel 1, Giza168 and Masr 1, respectively with 200
mg/l of ASA. On the other hand, data in Table (1) showed that shoot,
root length decreased in all wheat varieties with increasing of drought
stress by PEG. The highest values of shoot and root length were found in
Giza168 (11.33 cm) and (10.66 cm), respectively under normal
ASA PEG Germination % Shoot length (cm) Root length (cm)
mg/l bars
Shandaw
eel 1
Giza168 Masr 1
Shandaw
eel 1
Giza168 Masr 1
Shandaw
eel 1
Giza168 Masr 1
L.S.D 2.10 0.32 0.88
0.05
4.667
10.000
65.000
98.000 99.000
85.000 5.667
8.667
5.333
95.000
0
6.667
1 8.000 7.333
3
6.667
5.333
83.000
2
4.333
2.667
98.000 8.667
95.000
8.333
7.333
10.667
9.000
3 85.000 60.000 6.333 5.333 7.667 6.333
4.667 7.000
65.000 6.667 5.667 7.333
150
200
8.333
75.000
98.000
0.000
12.667 11.000
53.000
11.333
0.000
96.000 98.000 96.000 11.333
0
95.000
7.333
11.333 11.333 12.667
10.333
6.333
98.000
0
0.000
11.333
1 96.000
0.000
3.333
8.333
0.000
12.000
81.000
10.667
0.000
4.667
10.333
2 81.000
0.000
0.000
0.000 0.000 0.000
97.000
10.667
94.000 6.000
0 10.333
9.000
0.000
91.000
0.000 0.000
0.000
86.000
0.000
4.000
0.000
76.000 56.000
93.000 4.667
92.000 11.000
8.000 6.000
8.000
0.000
1.833
3
2
7.667
2.333 2.333 6.000 5.667
1
Egypt. J. of Appl. Sci., 36 (1) 2021 34
conditions. The lowest values of shoot and root length were observed in
Shandaweel 1 (1.83 cm) and (2.33 cm) respectively at 2 bar drought
stress. Application of ASA as soaking significantly increased shoot, root
length under drought and non-drought. Shoot length in shandaweel1
significantly increased from 6.0 cm to 6.66 cm at 1 bar and increased
from 1.83 cm to 2.66 cm at 2 bar with 150 mg/L of ASA. Also, 200 mg/L
of ASA significantly increase of shoot length in Giza 168 and Masr 1
from 4.0 and 2.33 cm to 6.66 and 7.33 cm, respectively at 2 bar. From the
results, it is obvious that ascorbic acid pretreatment alleviated the
inhibitory effect of drought stress on germination and growth parameters.
Also, 200 mg/L of ASA had higher effect compared to 150 mg/l of ASA.
Increased shoots and root length by ascorbic acid might be due to the cell
division and differentiation of meristem cells (Liso et al., 1998). These
results agreed with DolatAbadian and Sanavy (2008) reported that
priming with ascorbic increased germination percentage, Length of shoot
and root, their dry weight, and seedling total dry weight in sunflower.
The effect of ascorbic acid on photosynthetic pigments in shoots of
three wheat varieties under drought by PEG
Data presented in Table (2) showed that photosynthetic pigments
i.e. chlorophyll a, b and carotenoids were significantly decreased with
increasing of drought stress by PEG. chlorophyll a, b and carotenoids of
Giza 168 higher than Masr 1 and Shandaweel 1 under drought and control.
under control, the maximum values of chlorophyll a, b and carotenoids were
0.470,0.394 and 0.628 mg/g, respectively were found in Giza 168. Drought
stress led to significantly decreased chlorophyll a, b and carotenoids in all
wheat varieties. The lowest values of chlorophyll a was 0.293 mg/g in
shandaweel 1, while, the lowest values of chlorophyll b and carotenoids
were 0.193 and 0.187 mg/g, respectively in Masr 1 at 2 bar. Application of
ASA as soaking led to increase chlorophyll a, b and carotenoids contents
under drought stress. Chlorophyll a, b and carotenoids of Masr 1
significantly increased from (0.212, 0.194 and 0.298 mg/g) to (0.309, 0.255
and 0.389 mg/g), respectively with 150 mg/l of ASA and significantly
increased to (0.388, 0.323 and 0.562 mg/g), respectively with 200 mg/l of
ASA at 1 bar. The highest values of chlorophyll a, b and carotenoids (1.217,
0.842 and 1.079 mg/g) were found in Giza 168 with 200 mg/l of ASA under
normal conditions. At 3 bar, the chlorophyll a, b and carotenoids of Giza
168 were higher than Shandaweel 1 and Masr 1 with 200 mg/L of ASA.
From these results showed that Giza 168 had higher chlorophyll a, b and
carotenoids under drought stress and 200 mg/L is the most effective
concentration of ASA in reducing the effect of drought stress on total
pigment. The decrease of chlorophyll content under water limited condition
is reported to take place because of its photo-oxidation and degradation
under drought (Anjum et al., 2011). Exogenous application of ascorbic and
35 Egypt. J. of Appl. Sci., 36 (1) 2021
mitigated the adverse effects of drought on photosynthesis in all wheat
varieties by increasing stomatal conductance. This could have also been due
to the fact that ascorbic acid as an antioxidant has the ability to mitigate the
negative effects of stress on plants by neutralizing harmful oxidants which
have been reported to damage plant membranes such as the thylakoid
membranes of chloroplasts (Dolatabadian et al., 2009). These results agree
with Khalil et al. (2010) who found that the application of ascorbic acid in
different concentrations showed significant increase in all photosynthetic
pigments in Ocimun basilicum plant under water stress. Also, Malik and
Ashraf, (2012) found that Ascorbic acid treated seeding of both genotypes
maintained higher chlorophyll contents, net- photosynthesis and growthcompared
to the non-treated plants.
Table (2): The effect of ascorbic acid on photosynthetic pigments in
seedling of three wheat varieties under drought stress by
PEG.
ASA PEG Chlorophyll a (mg/g F. Wt) Chlorophyll b (mg/g F.Wt) Carotenoids (mg/g F.Wt)
mg/l bars
shandawe
el 1
Giza168 Masr 1
shandawe
el 1
Giza168 Masr 1
shandawe
el 1
Giza168 Masr 1
L.S.D 0.002 0.30 0.01
0.05
3
0.417
0.232
0.869 0.388
0.443
0.642
0.475
1.217
2
0.000 0.000
2 0.217
0
0.176
0.825
0.650
0.000
0.733
0.000
0.218
0.000
0.188 0.271
0.389
0.646
0.842
0.255
0.193
0.309
0.591
0.268 0.163
0.657
0.862 1.079
0.000
0.251
3
0.102
0.000 0.000 0.000 0.000
0.224
150
0.628
0.358
0
0
2
0.408
0.621
0.000
0.479
0.937
0.094
0.252
0.195 0.204
0.307 0.430 0.273
0.106
0.373
0.192 0.148
0.942 0.323 0.622 0.647 0.562
0
0.359
0.261 0.380 0.309 0.392 0.416 0.583
200
0.394
0.366 0.212 0.199 0.244 0.194 0.386
0.566
0.226 0.128
0.376
0.829
0.265
1
0.000
0.413 0.470 0.370 0.592 0.515
0.000
0.548
1
3
0.317
0.577 0.530
0.000 0.000 0.000 0.000
1 0.298
0.000
0.293 0.193 0.228
0.348
0.352 0.243 0.187
Egypt. J. of Appl. Sci., 36 (1) 2021 36
The effect of ascorbic acid on Catalase, Peroxidase and Proline in
seedling of three wheat varieties under drought stress by PEG
Results in Table (3) showed that catalase (CAT) activity
significant increase in Giza168 with increasing of drought stress at 3 bar,
whereas CAT activity significant increase in Masr 1 and nonsignificant
increase in Shandaweel 1 at 1 bar and then decreased at 2 bar. Under
control without ASA, the maximum CAT activity was observed in
Giza168 (40.33 U/mg F.W) at 2 bar and the minimum CAT activity was
found in Masr 1 (29.400 U/mg F.W) under normal conditions.
Pretreatment of with 150 mg/L of ASA caused significant increasing of
CAT activity in Masr 1 at all level of drought stress while not effective in
Giza168. 200 mg/L of ASA is the most effective in increase CAT activity
as compared to 150 mg/L (Table 3). CAT activity significant increase in
Masr 1 from 32.380 to 35.433 U/mg F.W with 150 mg/L of ASA and
significantly increased to 45.63 U/mg F.W with 200 mg/L of ASA at 2
bar. CAT activity in Shandaweel 1 decreased to 25.53 U/mg F.W with
150 mg/L of ASA, while significantly increased to 35.56 U/mg F.W with
200 mg/L of ASA at 2 bar.
Also, the results in Table (3) showed that peroxidase (POX)
activity significantly increased by increasing of drought stress at 3 bar in
all wheat varieties. Under control without ASA, the highest value of POX
activity was found in Shandaweel 1 (40.33 U/mg F.W) at 2 bar and the
lowest value of POX activity was found in Shandaweel 1 (23.36 U/mg
F.W) under normal conditions. Application of ASA as soaking caused
significantly increasing of POX activity in all wheat varieties (Table 3).
POX activity significantly increased in Giza168 from 30.40 U/mg F.W at
2 bar to 55.33 U/mg F.W with 150 mg/L of ASA and increased to 80.63
U/mg F.W with 200 mg/L of ASA at the same level of drought stress.
The maximum POX activity was observed in Shandaweel 1 (93.36 U/mg
F.W) and Masr 1 (92.40 U/mg F.W) at 3 bar with 200 mg/L of ASA. The
increased activities of CAT and POX led to limit cellular damage and
enhance the plants oxidative capacity to defend stress. CAT and POX
activities play a central protective role in the O2 and H2O2 scavenging
process (Hoque et al, 2007). The CAT and POX activity increased under
drought stress when compared to control plants. Similar results reported
under drought stress in wheat (Shao et al, 2005). These results are
similar to that obtained by Rezaei et al. (2013) showed that ascorbic acid
led to increasing of catalase, peroxidase activities Dracocephalum
moldavica L. under drought stress.
37 Egypt. J. of Appl. Sci., 36 (1) 2021
Table (3): The effect of ascorbic acid on Catalase, Peroxidase and
Proline in seedling of three wheat varieties under drought
stress by PEG
On the other hand, drought stress increased the proline contents in
all wheat varieties Table (3). Under control without ASA, a maximum
increase of proline contents was observed in Giza168 (13.96) mmoles/g
at 2 bar. Proline contents in Masr 1 was (12.20) mmoles/g higher than
Shandaweel 1 (10.36) mmoles/g at 2 bar. Application of ASA as soaking
significantly increased proline contents in all varieties (Table 3). The
highest value of proline was found in Giza168 (28.96) mmoles/g at 3 bar
with 200 mg/L of ASA. Application of 200 mg/L of ASA had higher
ASA PEG Catalase (U/mg F.W) Peroxidase (U/mg F.W) Proline (μmoles/g F.W)
mg/l bars
Shandaw
eel 1
Giza168 Masr 1
Shandaw
eel 1
Giza168 Masr 1
Shandaw
eel 1
Giza168 Masr 1
L.S.D 2.21 2.98
0.05
0.98
21.630
3 36.667 50.400 48.700 93.367 75.367 92.400 20.250 28.960 24.960
87.667 80.633 77.500 19.710 25.890
20.110
1 42.400 44.500 44.400 52.467 77.700 67.533 18.560 26.550 21.550
68.800 67.733 70.700 16.850 23.580
200
0 38.400 45.533 40.367
2 35.567 46.667 45.633
19.560
3 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000
66.633 55.333 65.667 17.740 21.500
10.290
1 35.467 38.700 36.567 52.400 52.300 41.333 15.600 18.960 15.690
40.367 44.500 35.533 10.850 12.570
150
0 32.667 38.400 30.467
2 25.533 40.667 35.433
12.200
3 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000
40.333 30.400 36.533 10.360 13.960
9.330
1 32.300 38.267 34.467 31.200 28.300 33.767 9.550 11.560 9.880
23.367 27.317 32.367 8.250 10.220
0
0 30.590 36.433 29.400
2 29.300 40.333 32.380
Egypt. J. of Appl. Sci., 36 (1) 2021 38
effect compared to 150 mg/L in increasing of proline contents in all
wheat varieties. For example proline contents significantly increased in
Shandaweel 1 from 10.36 mmoles/g at 2 bar to 17.74 mmoles/g with
150 mg/L of ASA while increased to 19.71 mmoles/g with 200 mg/L of
ASA. The role of proline as a protective agent against reactive oxygen
species is also very important (Hare et al., 1999). Drought tolerance is
positively correlated with high accumulation of proline in many crops
like wheat and barley (Nayyar and Walia, 2003). These results agreed
with Baghizadeh et al. (2009) who found that proline contents increased
with ascorbic acid under drought stress in Okra ( Hibiscus esculentus L.).
REFRENCES
Aebi, H. (1984): Catalase in vitro. Methods of Enzymology, 105:121-
126.
Afzal, I. ; S.M.A. Basra ; N. Ahmed and M. Farooq (2005): Optimiz
of hormonal priming techniques for alleviation of salinity
stress in wheat ( Triticum aestivum L.). Universidade de Santa
Cruz do Sul., 17(1):95-109.
Amako, A. ; K. Chen and K. Asada (1994): Separate assays specific
for ascorbiate peroxidase and gauiacol peroxidase and for
the chloroplast and cytosolic isoenxymese of ascorbate
peroxidase in plants. Plant cell physiol., 35:497-504.
Anjum, S.A.; X. Xie ; L. Wang ; M. F. Saleem ; C. Man and W. Lei
(2011): Morphological, physiological and biochemical
responses of plants to drought stress. African J. Agric. Res., 6:
2026-2032.
Baghizadeh, A.; M. Ghorbanli ; H.M. Rezaei and H. Mozafri (2009):
Evaluation of Interaction effect of drought stress with ascorbate
and salicylic acid on some of physiological and Biochemical
parameters in okra ( Hibiscus esculentus L.). Research Journal
of Biological Sciences, 4 (4): 380-387
Bates, L.S.; R.P. Walden and I.D. Teare (1973): Rapid determination
of free proline for water-stress studies. Plant Soil, 39: 205-207.
Bushuk, W. (1998): wheat breeding for end-product use Euphytica,
carbohydates and Na+/K+ changes in two tomato (lycopersicon
cell. plant physiol., 85: 529-536.
Dodd, G.L. and L.A. Donovan (1999): Water potential and ionic effects
on germination and seedling growth of two cold desert shrubs.
Am. J. Bot., 86:1146-1153.
39 Egypt. J. of Appl. Sci., 36 (1) 2021
Dolatabadian, A. and A.M. Sanavay (2008): Effect of the ascorbic
acid, pyridoxine and hydrogen peroxide treatments on
germination, catalase activity, protein and malonidialdehyde
content of three oil seeds. Bot . Hort . Agrobot . Cluj, 36 ( 2 ):
61 – 66.
Dolatabadian, A.; A.M. Sanavy and M.S. Harifi (2009): Effect of
ascorbic acid (Vitamin C) leaf feeding on antioxidant enzymes
activity, proline accumulation and lipid peroxidation of canola
(Brassica napus L.) under salt stress condition. Journal of
Science and Technology of Agriculture and Natural Resources
13: 611-620.
Esfandiari, E.; M.R. Shakiba ; S. Mahboob ; H. Alyari and M.
Toorchi (2007): water stress, antioxidant enzyme activity and
lipid peroxidation in wheat seedling. Journal of food
Agriculture α Environment, 5:149-153 .
Foyer, C.H. and G. Noctor (2003): Redox sensing and signaling
associated with reactive oxygen in chloroplast, peroxisomes
and mitochondria . physiol. Plant., 119: 355-364.
Gamze, O.; D.K. Mehmet and A. Mehmet (2005): Effect of salt and
drought stresses on germination and seedling growth of pea
(Pisum sativum L). Turk. J. Agric., 29: 237-242.
Gratao, P.L.; A. Polle ; P.J. Lea and R.A. Azevedo (2005): Making
the life of heavy metal – stressed plants a little easer.
Functional plant Biology, 32: 481 -494.
Hare, P.D.; W.A. Cress and J. Van Staden (1999): Proline synthesis
and degradation: a model system for elucidating stress-related
signal transduction. J. Exp. Bot., 50: 413-434.
Hoque, M.A.; E. Okcma ; M.N.A. Banu ; Y. Nakamura ; Y.
Shimoishi and Y. Murata (2007): Exogenous proline mitigates
the detrimental effects of salts tressmorethan exogenous betaine
by increasing antioxidant enzyme activities, Journal of Plant
Physiology, 164: 553-561.
ISTA, (1993): International Seed Testing Association. International
Rules For Seed Testing – Seed SCl, and Technol., 21:187-209.
Kavi Kishor, P.B.; S. Sangam ; R. N. Amrutha ; P. Sri Laxmi and
K.R. Naidu (2005): Regulation of proline biosynthesis,
degradation, up take and transport in higher plants: Its
implications in plant growth and abiotic stress tolerance. Curr.
Sci., 88: 424-438.
Egypt. J. of Appl. Sci., 36 (1) 2021 40
Khalil, S. E.; N.G. Abd El-Aziz and B. H. Abou Leila (2010): Effect
of water stress, ascorbic acid and spraying time on some
morphological and biochemical composition of ocimum
basilicum plant. Journal of American Science, 6 (12): 33-44.
Krishnasamy, V. and D.V. Seshu (1990): Germination after accelerated
ageing and associated characters in rice varieties. Seed Sci.
Technol., 18: 147-156.
Lichtenthaler, H.K. and A.R. Wellburn (1985): Determination of total
carotenoids and chlorophylls A and B of leaf in different
solvents. Biochem. Soc. Trans., 11: 591-592.
Liso, R.G.; A.M. Innocenti ; A. Bitonti and O. Arrigoni (1998):
Ascorbic acid-induced progression of quiescent center cells
from G1 to S phase, New Phytologist, 110: 469-471.
Malik, S. and M. Ashraf (2012): Exogenous application of ascorbic
acid stimulates growth and photosynthesis of wheat (Triticum
aestivum L.) under drought. Soil Environ., 31(1) :72-77.
Michel, B.E. (1983). Evaluation of the Water Potentials of Solutions of
Polyethylene Glycol 8000. Plant Physiol., 72:66–70.
Moaveni, P. (2011): Effect of water deficit stress on some physiological
traits of wheat (triticum aestivum). Agricultural science
Research Journal, 1:64-68.
Monakhova, O.F. and I.I. Chernyadev (2002): protective role of
kartolin-4 in wheat plants exposed to soil drought. Applied and
Environmental Microbiology, 38: 373-380.
Nayyar, H. and D.P. Walia (2003): Water stress induced proline
accumulation in contrasting wheat genotypes as affected by
calcium and abscisic acid. Biol. Plant., 46: 275-279.
Rezaei, H.; G. Mahlagh ; P. Maryam and A. Pazoki (2013): Effect of
drought Interactions with Ascorbat on some Biochemical
Parameters and Antioxidant Enzymes Activities in
Dracocephalum moldavica L. Middle-East J. Sci. Res., 13 (4):
522-531.
Shaddad, M.A ; A.F. Radi ; A.M. Abdel-Rahman and M.M. Azooz
(1990): Response of seeds of Lupinus termis and Vicia faba to
the interactive effect of salinity and ascorbic acid or pyroxidase.
Plant Soil, 122:177-183.
Shalata, A. and P.M. Neumann (2001): Exogenous ascorbic acid
(Vitamin C) increase resistance to salt stress and reduces lipid
peroxidation. Journal of experimental Botany, 52: 2207-2211.
41 Egypt. J. of Appl. Sci., 36 (1) 2021
Shao, H.B.; Z.S. Liang and M.A. Shao (2005): Changes of antioxidative
enzymes and MDA content under soil water deficit
among 10 wheat (Triticum aestivum L.) genotypes at maturation
stage, Colloid Surf Biointerf, 45: 7-13.
Sivakumar, P .; P. Sharmila and P. Pardha Saradhi (2000): proline
alleviates salt stress induced enhancement in ribulose 1,5-
bisphosphat oxygenase activity. Biochem Biophys. Res
Commu., 279:512-515.
Steel, R.G. and J.H. Torrie (1980). Analysis of covariance. Principles and
procedures of statistics: A Biometrical Approach, pp.401-437.
Tas, S. and B. Tas (2007): Some physiological responses of drought
stress in wheat genotypes with different ploidity in Turkiye.
World Journal of Agricultural Science, 3: 178-183.
Zhu, J.K. (2006): Effect of drought stresses induced by polyethylene glycol
on germination of pinus Silvestre's var. Mongolica seeds from
pollination forests on sandy land. Natural and pollination Forests
on sandy Land Journal of Forest Research, 11(5): 319-328.
تأثير المعاممة بحمض الاسکوربيک عمى باد ا رت القمح تحت إجهاد الجفاف
سعيد احمد خضر، سام محمد امين الحمامصى ، هيثم أحمد زکى الخميسى ،
زکريا حسن سعد
قسم الکيمياء الحيوية، کمية الز ا رعة، جامعة الأزهر، القاهرة، مصر.
تم تقييم ثلاثة أصناف من القمح المصري )شندويل 1، وجيزة 161 ، ومصر 1( تحت
3 بار( ، 2 ، 1 ، اجهاد الجفاف باستخدام ترکي ا زت مختمفة من البولي إيثيمين جلايکول ( 0
ود ا رسة تأثير تطبيق حمض الأسکوربيک بترکي ا زت ) 0 و 150 و 200 ممجم / لتر( في
التخفيف من إجهاد الجفاف في مرحمة الانبات لأصناف القمح. أوضحت النتائج أن إجهاد
الجفاف تسبب في انخفاض نسبة الإنبات ومقاييس النمو )طول النبات وطول الجذر( وصبغات
البناء الضوئي )الکموروفيل أ ، والکموروفيل ب ، والکاروتينات( مع زيادة ترکي ا زت البولى ايثمين
في (CAT) في جميع أصناف القمح. علاوة عمى ذلک، ا زد نشاط الکتاليز )PEG( جميکول
في صنف مصر CAT صنف جيزة 161 مع زيادة إجهاد الجفاف عند 3 بار ، بينما ا زد نشاط
1 و ا زد زيادة غير معنوية في صنف شندويل 1 عند 1 بار ثم انخفض عند 2 بار. أيضا، ت ا رکم
البرولين تحت ضغط الجفاف في جميع أصناف القمح. من ناحية أخرى، أدت معاممة البذور
بحمض الأسکوربيک قبل ز ا رعتها إلى تحسين جميع العوامل المدروسة تحت إجهاد الجفاف.
يعتبر 200 مجم / لتر من حمض الاسکوربيک هو الأکثر فاعمية في تخفيف آثار إجهاد الجفاف
عمى الخصائص المورفولوجية والکيميائية لأصناف القمح في مرحمة الأنبات.
Egypt. J. of Appl. Sci., 36 (1) 2021 42
