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Investigation of antioxidant system activity in durum wheat under drought tolerance.


Durum wheat were tetraploid species from wheat and after bread wheat ranked second place in terms of acreage ,durum wheat has been an important cereal in human nutrition which considered for produced the value that consists of semolina recently. Durum wheat to the viewpoint of having certain characteristics can be planted in different areas that planting bread wheat in such as climate in terms of difficulty, therefore durum wheat could cultivated in the soils low efficiency, under different environmental stress of such as cold, drought, heat, humidity and types of biotic and antibiotic stress [20,10,8]. In addition to morphological traits that plants adapted to tolerate stress conditions is involved in physiological indices of plant breeding to select superior genotypes to unfavorable environmental conditions is very important.

Selection of genotypes tolerant to stress conditions, methods and different traits can be considered that the review indicators and quantitative traits among these has special significance [19,15,17,3].

When plants are placed in water stress conditions be low water of inside the plant tissue, small cell especially and osmotic pressure inside the tissue is much finally, and some of the available oxygen in the cells lose their electrons and become free radicals of oxygen, active oxygen converted electrons in normal conditions and change to water, but active oxygen damage to plant tissue in terms of stress.

Many physiological reactions and oxidase enzymes produced active oxygen in plant cells, which are the reactions of plant responses to environmental stress [5].

Plant defense systems to reduce the free oxygen in the cellular tissue and repair damaged tissue often can be in the role of antioxidant enzymes superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, glutathione peroxidase, dihydro ascorbat reductase [14]. Redoxin proxy react with hydrogen peroxides in chemical reactions inside the cell and hydrogen peroxides will become water molecules, and thereby this chemical composition inhibited free radicals into cell with putting hydrogen atoms, and provide of plant resistance against stress factors. Redoxin proxy oxidizes and creating feudal union between the two molecules cause of changing oxide condition to the dioxide position, which done by Tioredoxin for defense mechanisms in reducing free radical acts.

Redoxin proxy reacting at the mitochondria, cytosell, chloroplast organs [16]. Superoxide are a group of metallo-enzymes, its role change free oxygen (O2) to the molecular oxygen and hydrogen peroxides, This enzyme plays an important role in reducing the active oxygen and had a plant defense mechanisms against environmental stress. Enzyme superoxide disamutase are three groups ,one group wealthy with manganese metals and considered manganese possessor superoxide disamutase or Mn-SOD. Enzyme ascorbate peroxidase contain the most important role in the removal of hydrogen peroxide, which Enzyme ascorbate peroxidase prevents injury of effects oxidation stress, Enzyme ascorbate exist in chloroplast, cytosell, vacuole and ecoplast space of leaf cell in different concentrations [9]. Oxidation of ascorbate peroxidase doing the following position, that the first process ascorbate change to the monohydroascorbat then revival to the ascorbate or change to the ascorbate and dihydroascorbat.


Ascorbate was more chloroplast sytosell ascorbate peroxidase activity, but in some cases have been reported in mitochondria [1]. Cellular defense mechanisms arise of the combined activities of several enzymes that harmony and balance between them is very important drought resistance in plant. Disorder in any of the defense system affected to the other systems, So for example if decreases the enzyme activity Superoxide, this phenomenon increased the radicals superoxide dismutase in cells and caused increased radical reactions Hber-wis, and arise super devastating radical of hydroxyl type to cells. In addition increased activity of Superoxid dismutase radicals and Proxid hydrogen reduced activity of other important defense enzymes, including catalase and peroxidase [7,3]. Antioxidants were reduced active oxygen's by analysis into cell, and retinued equilibrium physiological of cells, so increased resistance of plant in the stress condition. Each activity was reduced with increased antioxidant enzyme levels in the cell is compensated in some cases even as antioxidants act to antioxidant coenzyme [13]. Different plants have antioxidant systems such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR) and some antioxidants low weight molecular are non-enzyme, that antioxidant systems reduce cell oxidizing and free radicals against environment stress and prevent cytoplasmic membranes degradation , most of these enzymes are detectable with laboratory methods. The Enzymes can be resistance for stress situation are used as a criterion for selecting resistant cultivars to the stress environmental conditions [12,3].

Material and methods

Experiments tested basic design of agricultural experiments and templates as a randomized complete block split plot with three replications and tow environments of water stress and normal since 2010 in Ardebil branch Islamic Azad University of Iran, Experiments performance with 12 varieties of durum wheat including ,Fadda , Yavarus, Altar, Omrabi5, Zardak, Stork Korifera, Vadalmez, Omrabio Hurani, Chakmag and Ammar in sub-plots. Measurements worked after of 12 days from applying dry randomly, and measured about 10 flag leaves per genotype and considered their average. number of experimental units was 72 and area per unit test was approximately 3 [m.sup.2], sample was performed after removing marginal effects. All agricultural operations perform as a common and uniformly in stress and normal conditions, and for irrigation stress delete the final step of irrigation. Leaves obtained from plants grown transferred to the laboratory and using liquid nitrogen for mortar and stored sample in the refrigerator. Enzymes ascorbate peroxidase (APO) extracted by sayram method [18] catalase (CAT), Superoxide dismutase (SOD) by Reye's and Gyanopolotys method [11] and extracted catalase enzyme by chance and Maehly method [6]. 5.0 grams of wheat leaf powder mixed with 10 ml of potassium buffer and centrifuged for 15 minutes with 4 [C.sup.0] temperatures and 20000 around to measure the ascorbate enzyme, 3 ml ascorbate peroxidase reaction was contained ,50 mmol of potassium buffer with 7 PH, .05 mmol ascorbic acid, a tenth mmol EDTA, and a tenth of ml enzyme and a tenth mmol hydrogen peroxidase.

Reaction starts by adding H2O and read by spectrophotometer at a wavelength of 290 nm immediately after adding H2O2 ,and second readings start after 60 seconds. Differences of reading will show amount of reduce, readings were calculated with ascorbic acid as standard regression curves. During the projects for analysis of variance and comparisons average in the results was used common statistical software SPSS, MSTATC, SAS, EXCEL.


According to variance analysis table are split plot see that the main plot factor A related to main plot is significant at 5%, and this affair shows that water stress and normal condition had significant effect in the performance of genotype. In examining the effect of factor B related to the durum wheat cultivars factor B has been significant at 1% probability level and this suggests showed that different cultivars have different yield. In these circumstances, as noted in the table1, effects of block are no significant and this affect has been shown that the various block related to experiments have balanced yield. Biomass yield showed significant effect at 1% level in all source of variation and Superoxide Dismutase (SOD), Catalase (CAT), ascorbic acid showed nonsignificant effect in same level, reason this case being for annual , therefore we use Duncan test for obtain subtle result in this case. Injury Index (Id)leaf tissue showed significant effect at 1% and this result ditinct that genotype had difference Injury Index. Relative water content(RWC) trait had significant effect at 5% level and ditinct RWC variation between genotypes. Mean comparisons treatments of grain yield , biomass , Superoxide Dismutase (SOD), Catalase (CAT), ascorbic acid , injury Index (Id) of leaf tissue and relative water content(RWC) were performed based on the least significant difference test (LSD) at 5% level in split-plot design experiments for gruping and fine result in normal and water stress (Table 2). genotype1 had highest yield amount 5037 kilograms of seed per hectare in the normal condition according to this grouping located A group, genotype 4 had the lowest yield in this grouping and located group L. genotype number of 7 with 6344 kilogram per hectare grain had second yield located in the group B. Stress conditions study in this experiment, cultivars of 10 amount 2506 kilogram of seed per hectar was the greatest yield located group A and cultivar of 7 placed group B and cultivar of 6 had lowest yield be group L in the water stress condition and the rest of the genotype were different groups according to table 2.

Comparison of biomass showed that genotype 10 has the highest yield per both normal and water stress conditions was placed in Group A and genotype No. 3 had the lowest biomass in normal and water stress conditions and genotype of 6 were produced lowest biomass in the stress conditions (table2). Result of this study showed that genotype number of 2 had the lowest percentage of Injury Index (Id) leaf tissue in water stress and normal conditions and grouped in I and J respectively. Relative water content (RWC) were study in water stress and normal condition and ditinct that genotype of 11 included the highest relative water content tissues in group A and other genotype were in group B in terms of the normal conditions, but genotype of 4 had the highest RWC include Group A in water stress conditions and number 9 was be the lowest RWC was including group D and genotype of number 10 featured second percent of RWC in water stress condition(Table 2).

Mean comparisons treatments of unit enzymes SOD, CAT, ascorbic acid , were performed based on LSD at 5% level showed that different action genotype per unit enzymes in normal and water stress conditions. Genotype number of one had the most enzyme superoxide dismutase units (SOD) and catalase (CAT) in water stress conditions, and was one of the resistant genotype related to drought conditions (Table 2).Correlation analysis was used between traits by Pearson method for obtain relation yield traits for grain of kilograms per hectare (Grain), biomass kilograms per hectare (Biomass), Relative water content (RWC), Injury index leaf tissue (Id), ascorbic acid enzyme unit (Ascorbic), unit of enzyme catalase (CAT), superoxide dismutase (SOD)in water stress and normal conditions (table 3,4,5).

Between trait grain yield per hectare and the amount of biomass was found positive significant relationship at 1% probability level in the stress conditions (Table 5). The review of these factors between grain yield with biomass, and between RWC with biomass and between Injury index (Id) with RWC, and between SOD with CAT was found significant relation at 5% level while between SOD with Injury index leaf tissue (Id) be significant relation at 1% level (table 4). In addition between average of two normal and water stress conditions studies relationship of traits, and found positive significant relationship at 5% level, between grain yield with biomass, and between ascorbic acid with RWC, and between SOD with Injury index leaf tissue (Id) in average both normal and water stress conditions (table3).


According to the results of performance reviews and related traits in the two conditions of water stress and normal conditions, and considering of testing and analysis of variance in split-plot design, and review of main plots for water stress and normal conditions, and study of sub plot effects for genotypes were observed significantly effect in the most of traits.

It is shown that drought-related indices have different yield in different genotypes which these results are according to results of other researchers [2,19]. In addition to analysis of variance using different statistical method for evaluation of graphs from the results of the tests also, Figure1 showed yield of grain per hectare in normal and water stress condition, genotype number 1 had the highest yield in normal condition, but genotype number 10 include highest yield in water stress conditions, The conclusion that the genotype number 1,2,3,7,11 are resistant cultivar for the drought conditions in this excrement. It was concluded that the antioxidant parameters can be used as an indirect selection criterion for screening drought-resistant of germ plasmas.



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(1) Hasan Bigonah Hamlabad, (2) Ibrahim Azizov, (3) Hamlet Sadiqov, (4) Zeynal Akparov, (5) Mostafa Valizadeh, (6) Hossein Shahbazi

(1,5,6) Ardabil branch, Islamic Azad University, Ardabil, Iran.

(2,3,4) Genetic Resources Institute, Azerbaijan National Academy of Sciences, Baku, Azerbaijan.

Corresponding Author

Hasan bigonah hamlabad, Ardabil branch, Islamic Azad University, Ardabil, Iran.

Mobile: 00989141568591.

E-mail: and
Table 1: Split-plot design analysis of variance.

S.O.V      df     yield         Biomass       Catalase      Ascorbic

Block      2      0.207 ns      1972 **       0.039 ns      0.019 ns
a          1      1.73 *        4329 **       1.91 ns       0.018 ns
Error1     2      0.128         2.37          0.028         0.006
b          11     0.503 **      1028 **       0.156 ns      0.0051 ns
ab         11     0.094 ns      200 ns        0.022 ns      0.0047 ns
Error2     44     0.166         427.01        0.122         0.0052
% CV              12.96         19.82         22.67         22.03

           Injury                      Water
S.O.V      Index         SOD           Content

Block      0.929 ns      1.29 ns       126.2 ns
a          18.8 ns       1.92 ns       940.62 *
Error1     5.27          1.013         84.45
b          6.03 **       0.102 ns      31.45 ns
ab         1.34 ns       0.07 ns       34.76 ns
Error2     1.25          0.168 ns      55.29
% CV       22.38         24.15         12.16

**--Significant at 1% and ns non significant

Table 2: Mean comparisons treatments of grain and biomass yield
per hectare in normal and water stress conditions.

            Grain kg/h                    Biomas kg/h

Genotype    N              S              N              S

1           5037 A         1903.63 D      16416.67 B     12166.67 C
2           4126.25 C      2249.47 C      10472.22 J     8347.22 I
3           1620.99 J      1035.58 I      9388.89 L      8986.11 G
4           882.44 L       469.29 K       9979.17 K      8152.78 J
5           3003.03 G      1801.21E       13263.89 E     12694.44 B
6           1061.72 K      448.92 L       11625.00 I     6097.22 L
7           4463.25 B      2252.75 B      15027.78 C     9638.89 E
8           3375 D         470.33 J       13333.33 D     9222.22 F
9           2326.08 H      1256.44 G      11875.00 H     10708.33 D
10          3246.64 F      2506.19 A      18458.33 A     17805.56 A
11          1667.25 I      1254.72 H      12861.11 F     6291.667 K
12          3254.11 E      1299.19 F      12777.78 G     8805.557 H

            RWC                           Id

Genotype    N              S              N              S

1           62.263 B       56.843 ABCD    33.58 C        35.06 E
2           63.533 B       60.01 ABC      5.20 J         16.34 I
3           67.23 AB       58.447 ABC     16.70 G        20 G
4           64.28 B        63.177 A       23.51 D        36.75 D
5           62.447 B       53.63 CD       7.78 I         39.27 C
6           67.273 AB      54.477 BCD     42.61 A        44.25 B
7           63.557 B       59.64 ABC      14.27 H        18.89 H
8           65.27 AB       56.42 ABCD     20.67 E        36.11 D
9           66.033 AB      50.143 D       32.97 C        38.88 C
10          62.013 B       61.093 AB      16.10 G        27.25 F
11          71.537 A       59.75 ABC      36.95 B        49.37 A
12          61.927 B       55.987 BCD     17.68 F        27.48 F

            Ascorbic                      CAT

Genotype    N              S              N              S

1           0.09 A         0.11 A         2.34 AB        3.86 A
2           0.10 A         0.15 A         1.87 CD        2.92 D
3           0.10 A         0.11 A         2.20 ABC       3.19 CD
4           0.10 A         0.17 A         2.39 A         2.85 D
5           0.09 A         0.11 A         2.05 ABC       2.22 E
6           0.08 A         0.11 A         1.97 BCD       3 D
7           0.10 A         0.11 A         2.22 ABC       3.11 CD
8           0.10 A         0.11 A         2.04 ABC       3.64 AB
9           0.09 A         0.13 A         1.92 CD        2.25 E
10          0.08 A         0.11 A         1.87 CD        2.82 D
11          0.10 A         0.14 A         2.25 ABC       3.41 BC
12          0.10 A         0.11 A         1.58 D         1.81 F


Genotype    N              S

1           2.716 A        4.746 A
2           1.821 D        3.903 BC
3           2.04 CD        4.074 BC
4           2.926 A        3.268 D
5           1.831 D        2.532 E
6           2.937 A        4.545 A
7           2.299 BC       2.614 E
8           2.723 A        3.03 D
9           2.593 AB       3.848 BC
10          2.748 A        4.191 B
11          2.939 A        4.075 BC
12          1.999 CD       3.802 C

Table 3: Correlation analysis between traits in average normal and
stress treatments.

          Grain                       Injury
          yield     Biomass  RWC      Index     Ascorbic  CAT      SOD

Grain     1

Biomass   0.628 *   1

RWC       -0.33     -0.289   1

Injury    -0.543    -0.179   0.088    1

Ascorbic  -0.311    -0.57    0.585 *  -0.086    1

CAT       0.027     -0.024   0.523    0.241     0.012     1

SOD       -0.256    0.043    0.286    0.651 *   -0.066    0.358    1

Table 4: Correlation analysis between traits in normal treatments.

          Grain                       Injury
          yield     Biomass  RWC      Index     Ascorbic  CAT      SOD

Grain     1

Biomass   0.596 *   1

RWC       -0.626 *  -0.429   1

Injury    -0.414    0.008    0.577 *  1

Ascorbic  0.076     -0.476   0.166    -0.344    1

CAT       -0.135    -0.063   0.313    0.257     0.205     1

SOD       -0.374    0.233    0.417    0.792 **  -0.328    0.407 *  1

Table 5: Correlation analysis between traits in stress treatments.

          Grain                       Injury
          yield     Biomass  RWC      Index     Ascorbic  CAT      SOD

Grain     1

Biomass   0.631 *   1

RWC       0.178     0.005    1

Injury    -0.521    -0.222   -0.348   1

Ascorbic  -0.187    -0.363   0.449    0.105     1

CAT       -0.083    -0.128   0.365    0.073     -0.025    1

SOD       -0.027    -0.03    -0.019   0.14      -0.04     0.263    1
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Title Annotation:Original Article
Author:Hamlabad, Hasan Bigonah; Azizov, Ibrahim; Sadiqov, Hamlet; Akparov, Zeynal; Valizadeh, Mostafa; Shah
Publication:Advances in Environmental Biology
Article Type:Report
Geographic Code:7IRAN
Date:Jul 1, 2011
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