Drought induced programmed cell death and associated changes in antioxidants, proteases, and lipid peroxidation in wheat leaves

Two wheat (Triticum aestivum L.) genotypes with varying degree of drought tolerance were used to analyze programmed cell death (PCD) and related biochemical changes under drought stress. Drought induced PCD in leaves, as evident by internucleosomal nDNA fragmentation, was observed in sensitive genotype Nesser. Drought tolerant genotype (FD-83) showed higher peroxidase, superoxide dismutase, and catalase activities and ascorbate content under drought stress compared to sensitive genotype. Total phenolic content increased whereas lipid peroxidation remained un-changed under drought in FD-83. In contrast, drought enhanced the proteases and ascorbate peroxidase activities and lipid peroxidation (MDA content) in Nesser.     

Polyamine catabolism influences antioxidative defense mechanism in shoots and roots of five wheat genotypes under high temperature stress

Effect of high temperature stress on polyamine catabolism and antioxidant enzyme activity in relation to glutathione, ascorbate and proline accumulation was studied in five wheat (Triticum aestivum L.) genotypes (differently susceptible to temperature stress). High temperature significantly increased the activities of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and glutathione S-transferase (GST) in shoots of all genotypes. Higher activities of GPX in C 306, C 273 and APX in PBW 550, PBW 343 and PBW 534 demonstrate their important role in scavenging H₂O₂. Conversely, high temperature stress led to a significant decline in SOD, CAT, APX and GPX activities of roots with a subsequent increase in diamine oxidase (DAO) and polyamine oxidase (PAO) activities especially in PBW 550 and PBW 343. The concentration of ascorbic acid declined with the imposition of heat stress, however, polyamines responded to high temperature stress by increasing spermidine and spermine levels and decreasing putrescine levels. After exposure to high temperature, proline accumulation was significantly decreased in roots and increased in shoots though maximum concentration was achieved in C 306 genotype. Apparently, the wheat seedlings respond to high temperature mediated increase in reactive oxygen species (ROS) production by altering antioxidative defense mechanism and polyamine catabolism though differentially in five wheat genotypes. Among five genotypes studied, C 306 and C 273 seem to be better protected against temperature stress. The results suggested that shoots were more resistant against the destructive effects of ROS as is indicated by low levels of thiobarbituric acid reactive substances under high temperature stress.     

Effect of short-term heat stress on growth, physiology and antioxidative defence system in wheat seedlings

An experiment was conducted to find out the effect of short-term heat stress on morpho-physiological characters and antioxidants in 10 diverse wheat genotypes. Seed were aseptically grown in test tubes containing filter paper whose lower half was dipped in one-fourth MS media. Heat stress conditions were created by exposing the seedlings at 45 °C for 2 h after 7 days of their germination. Measurements were taken after 3 days of treatment. Heat stress significantly reduced the shoot dry mass, root dry mass, shoot length and root length in all the genotypes. The chlorophyll content and membrane stability index decreased, whereas proline content increased in heat-treated plants. There was significant increase in the activity of catalase, guaiacol peroxidase and superoxide dismutase under stress conditions. The genotypic variations were also significant. On the basis of a coordinated simulation of all these parameters, wheat genotypes Raj 4037 and PBW 373 were identified as tolerant to high temperature stress. The study provides evidence that the tolerant genotypes were equipped with better management of physiological processes along with an efficient antioxidative defence system, sensitivity of which can be evaluated to a sufficient level of certainty at seedling stage.     

Evaluation of oxidative stress tolerance in maize (Zea mays L.) seedlings in response to drought

Seedlings of selected six genotypes of maize (Zea mays L.) differing in their drought sensitivity (LM5 and Parkash drought-tolerant and PMH2, JH3459, Paras and LM14 as drought-sensitive) were exposed to 72 h drought stress at two leaf stage. Alterations in their antioxidant pools combined with activities of enzymes involved in defense against oxidative stress were investigated in leaves. Activities of some reactive oxygen species (ROS)-scavenging enzymes, catalase (CAT) and ascorbate peroxidase (APX) were enhanced in tolerant genotypes in response to drought stress. Superoxide dismutase (SOD) activity was significantly decreased in sensitive genotypes, but remained unchanged in tolerant genotypes under stress. Peroxidase (POX) activity was significantly induced in tolerant, as well as sensitive genotypes. Imposition of stress led to increase in H2O2 and malondialdehyde (MDA, a marker for lipid peroxidation) content in sensitive genotypes, while in tolerant genotypes no change was observed. Significant increase in glutathione content was observed in sensitive genotypes. Ascorbic acid pool was induced in both tolerant and sensitive genotypes, but induction was more pronounced in tolerant genotypes. Significant activation of antioxidative defence mechanisms correlated with drought-induced oxidative stress tolerance was the characteristic of the drought tolerant genotypes. These studies provide a mechanism for drought tolerance in maize seedlings.     

Role of Antioxidant Systems in Wheat Genotypes Tolerance to Water Stress

The role of plant antioxidant systems in stress tolerance was studied in leaves of three contrasting wheat genotypes. Drought imposed at two different stages after anthesis resulted in an increase in H2O2 accumulation and lipid peroxidation and decrease in ascorbic acid content. Antioxidant enzymes like superoxide dismutase, ascorbate peroxidase and catalase significantly increased under water stress. Drought tolerant genotype C 306 which had highest ascorbate peroxidase and catalase activity and ascorbic acid content also showed lowest H2O2 accumulation and lipid peroxidation (malondialdehyde content) under water stress in comparison to susceptible genotype HD 2329 which showed lowest antioxidant enzyme activity and ascorbic acid content and highest H2O2 content and lipid peroxidation. HD 2285 which is tolerant to high temperature during grain filling period showed intermediate behaviour. Superoxide dismutase activity, however, did not show significant differences among the genotypes under irrigated as well as water stress condition. It seems that H2O2 scavenging systems as represented by ascorbate peroxidase and catalase are more important in imparting tolerance against drought induced oxidative stress than superoxide dismutase alone. This revised version was published online in July 2006 with corrections to the Cover Date.     

Drought tolerance in cereals in terms of water retention,photosynthesis and antioxidant enzyme activities

Experiments were carried out on three bread wheat varieties, one barley and one durum wheat variety grown in pots in the phytotron and subjected to water withdrawal for 7 days during grain-filling. Leaf water loss, net assimilation rate and transpiration showed marked differences, allowing the genotypes to be ranked. Although the most resistant variety had the highest activity for ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and glutathione-S-transferase (GST), which did not rise further in response to drought and the most susceptible variety had the lowest values, which increased to the greatest extent under drought, the level of sensitivity could not be predicted for all the genotypes from the enzyme activity values alone. The largest increases were recorded for the APX, CAT and GR activities. In most genotypes the GR activity was correlated with that of GST, CAT and APX. Changes in the enzyme activities were observed after a decline in transpiration and photosynthesis. The range of soil moisture values over which the antioxidant enzyme activity levels remained relatively unchanged was a better indication of tolerance to drought than either basic or stress-induced activity levels.     

Salinity induced oxidative stress and antioxidant system in salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.)

Indices of oxidative stress viz., superoxide radical and H₂O₂ content increased in leaves of all the cultivars with the rise in salinity level, the increase was more pronounced and significant in salt-sensitive varieties and non-significant in resistant cultivars. Except for glutathione reductase (GR), basal activities of all other antioxidative enzymes viz. superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione reductase (GR) were significantly higher in leaves of all the resistant cultivars as compared to the sensitive ones. A differential response of salinity was observed on various enzymatic and non-enzymatic components of antioxidant system in leaves of salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.). Activities of superoxide dismutase and glutathione reductase enhanced in all the tolerant cultivar while declined in the sensitive cultivars with increasing salinity from 0 to 100 mM. Salt-stress induced the activities of catalase and peroxidase in all the cultivars but the magnitude of increase was more pronounced in the sensitive cultivars than in the tolerant cultivars. Contrarily, APX activity increased in the salt-sensitive cultivars but showed no significant change in the salt-tolerant cultivars. The amount of ascorbic acid content, reduced glutathione (GSH), reduced/oxidized glutathione (GSSG) ratio was higher in leaves of the tolerant cultivars than that of the sensitive cultivars under saline conditions. It is inferred that leaves of salt-tolerant cultivars tend to attain greater capacity to perform reactions of antioxidative pathway under saline conditions to combat salinity-induced oxidative stress.     

Influence of abiotic stresses on the antioxidant enzyme activity of cereals

The unfavourable effects of climate change were studied in terms of changes in the stress tolerance of cereals. Changes in the antioxidant enzyme activities were analysed as a function of the weather in a 2-year field experiment in order to determine the effect of extreme temperatures and rainfall conditions on the enzyme activity. The enzyme responses of two winter wheat genotypes to drought stress, simulated by withholding water completely for 7 days, were analysed under phytotronic conditions in three phenophases. The plants were raised either at ambient CO₂ concentration or at a doubled level. The quantities of glutathione reductase (GR), glutathione-S-transferase (GST), catalase (CAT), guaiacol peroxidase (POD) and ascorbate peroxidase (APX) were determined from leaf samples. The peroxidases had the most intense activity during the winter and early spring periods, with guaiacol peroxidase being dominant until the end of the winter. CAT generally became more active in late spring and summer, the activity being correlated with the development of water deficiency. The activity of GR, GST, POD and CAT was found to increase during the dry period, while the role of GR and POD was extremely important for resistance to low temperature.     

Proline improves copper tolerance in chickpea (Cicer arietinum)

The present study suggests the involvement of proline in copper tolerance of four genotypes of Cicer arietinum (chickpea). Based on the data of tolerance index and lipid peroxidation, the order for copper tolerance was as follows: RSG 888?>?CSG 144?>?CSG 104?>?RSG 44 in the selected genotypes. The basis of differential copper tolerance in chickpea genotypes was characterized by analyzing, antioxidant enzymes (superoxide dismutase, ascorbated peroxidase and catalase), phytochelatins, copper uptake, and proline accumulation. Chickpea genotypes showed stimulated superoxide dismutase activity at all tested concentrations of copper, but H₂O₂ decomposing enzymes especially; ascorbate peroxidase did not increase with 25 and 50 μM copper treatments. Catalase activity, however, increased at lower copper concentrations but failed to stimulate at 50 μM copper. Such divergence in responses of these enzymes minimizes their importance in protecting chickpea against copper stress. The sensitive genotypes showed greater enhancement of phytochelatins than that of tolerant genotypes. Hence, the possibility of phytochelatins in improving copper tolerance in the test plant is also excluded. Interestingly, the order of proline accumulation in the chickpea genotypes (RSG 888?>?CSG 144?>?CSG 104?>?RSG 44) was exactly similar to the order of copper tolerance. Based on hyperaccumulation of proline in tolerant genotype (RSG 44) and the reduction and improvement of lipid peroxidation and tolerance index, respectively, by proline pretreatment, we conclude that hyperaccumulation of proline improves the copper tolerance in chickpea.     

Differences in physiological and biochemical characteristics in response to single and combined drought and salinity stresses between wheat genotypes differing in salt tolerance

The combined drought and salinity stresses pose a serious challenge for crop production, but the physiological mechanisms behind the stresses responses in wheat remains poorly understood. Greenhouse pot experiment was performed to study differences in genotype response to the single and combined (D + S) stresses of drought (4% soil moisture, D) and salinity (100 mM NaCl, S) using two wheat genotypes: Jimai22 (salt tolerant) and Yangmai20 (salt‐sensitive). Results showed that salinity, drought and/or D + S severely reduces plant growth, biomass and net photosynthetic rate, with a greater effect observed in Yangmai20 than Jimai22. A notable improvement in water use efficiency (WUE) by 239, 77 and 103% under drought, salinity and D + S, respectively, was observed in Jimai22. Moreover, Jimai22 recorded higher root K⁺ concentration in drought and salinity stressed condition and shoot K⁺ under salinity alone than that of Yangmai20. Jimai22 showed lower increase in malondialdehyde (MDA) accumulation, but higher activities of superoxide dismutase (SOD, EC 1.15.1.1) and guaicol peroxidase (POD, EC 1.11.1.7), under single and combined stresses, and catalase (CAT, EC 1.11.1.6) and ascorbate peroxidase (APX, EC 1.11.1.11) under single stress. Our results suggest that high tolerance of Jimai22 in both drought and D + S stresses is closely associated with larger root length, higher Fv/Fm and less MDA contents and improved capacity of SOD and POD. Moreover, under drought Jimai22 tolerance is firmly related to higher root K⁺ concentration level and low level of Na⁺, high‐net photosynthetic rate and WUE as well as increased CAT and APX activities to scavenge reactive oxygen species.     

Nitric oxide improves thermotolerance in spring maize by inducing varied genotypic defense mechanisms

The investigation aimed at determining the effect of nitric oxide on antioxidant defense system of spring maize (Zea mays L.) genotypes namely, LM 11 (stress susceptible) and CML 32 (stress tolerant), that showed differential tolerance towards heat stress. Seed priming with a NO donor, sodium nitroprusside (SNP) improved seedling growth and induced varied defense mechanisms, under stress conditions. 75 μM SNP improved seedling lengths and their biomasses. It specifically enhanced catalase (CAT) activity in the roots of stressed seedlings of the two genotypes. However, it could induce CAT activity only in LM 11 shoots, under heat stress. It also enhanced peroxidase (POX) activity in CML 32 roots. However, such induction of POX activity with SNP treatment was not observed in LM 11 roots. This showed that NO increased the H₂O₂ scavenging efficiency of CML 32 genotype by enhancing the cumulative activation of CAT and POX in its roots. However, it did not induce activation of any of the H₂O₂ detoxifying enzymes in CML 32 shoots which showed that ascorbate–glutathione cycle remained non-operational in shoots of SNP-treated seedlings of the tolerant genotype, under high temperature stress. With seed priming, superoxide dismutase (SOD) activity increased in both the tissues of LM 11 seedlings. The shoots of SNP primed CML 32 seedlings, however, did not show any effect on SOD activity which illustrated that nitric oxide might act as a direct scavenger of superoxide radicals in CML 32 seedlings. SNP decreased the contents of H₂O₂ and MDA and increased proline content in seedlings of both the genotypes indicating reduced oxidative damage. The results thus showed that nitric oxide might induce different mechanisms of stress tolerance in these maize genotypes.     

The ecotoxicological and interactive effects of chromium and aluminum on growth,oxidative damage and antioxidant enzymes on two barley genotypes differing in Al tolerance

The effects of single or combined stress of aluminum (Al) and chromium (Cr) on plant growth, root dehydrogenase, oxidative stress and antioxidative enzymes were studied using two barley genotypes differing in Al tolerance in a hydroponic experiment. Al or Cr stress decreased plant growth, lowered root dehydrogenase activity and caused oxidative damage, as characterized by increased MDA and H₂O₂ contents. Under Al or Cr stress, the activities of antioxidative enzymes, including superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT), were dramatically increased in plant tissues. Gebeina, an Al-tolerant genotype, had less oxidative damage than Shang 70-119, an Al-sensitive genotype. The extent of oxidative damage induced by Cr varied with the pH of the culture solution, with lower pH values (4.0) being more severe than higher pH values (6.5). The combination of Cr and Al caused a further decrease in plant growth, a decrease in root dehydrogenase activity and an increase in MDA and H₂O₂ contents as well as the activities of antioxidative enzymes. There was also a marked difference between the two barley genotypes in the extent of increased antioxidative enzyme activity under the Cr and Al stresses.     

Role of antioxidant and anaerobic metabolism enzymes in providing tolerance to maize (Zea mays L.) seedlings against waterlogging

The present investigation was undertaken to identify the possible mode of mechanism that could provide tolerance to maize (Zea mays L.) seedlings under waterlogging. Using cup method, a number of maize genotypes were screened on the basis of survival of the seedlings kept under waterlogging. Two tolerant (LM5 and Parkash) and three susceptible (PMH2, JH3459 and LM14) genotypes were selected for the present study. Activities of antioxidant and ethanolic fermentation enzymes and content of hydrogen peroxide (H2O2), glutathione and ascorbic acid were determined in roots of these genotypes after 72 h of waterlogging. Waterlogging treatment caused decline in activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in all the genotypes. However, only susceptible genotypes showed slight increase in glutathione reductase (GR) activity. Significant reduction in APX/GR ratio in susceptible genotypes might be the cause of their susceptibility to waterlogging. The tolerant seedlings had higher GR activity than susceptible genotypes under unstressed conditions. Stress led to decrease in H202 and increase in glutathione content of both tolerant and susceptible genotypes, but only tolerant genotypes exhibited increase in ascorbic acid under waterlogging conditions. In the tolerant genotypes, all the enzymes of anaerobic metabolism viz. alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and pyruvate decarboxylase (PDC) were upregulated under waterlogging, whereas in susceptible genotypes, only ADH was upregulated, suggesting that efficient upregulation of entire anaerobic metabolic machinery is essential for providing tolerance against waterlogging. The study provides a possible mechanism for waterlogging tolerance in maize.     

Changes in some anti-oxidative enzymes and physiological indices among sesame genotypes (Sesamum indicum L.) in response to soil water deficits under field conditions

A field experiment was conducted to evaluate the response of ten sesame genotypes to different levels of soil water in terms of contents of proline, soluble carbohydrates, carotenoids, and activities of catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APX). Plants were grown under three irrigation levels, including irrigation at 55 % (control), 75, and 85 % depletion of soil available water. Field test plots were a two-way factorial arranged in a randomized complete block design with three replications. Under control level of irrigation, the most and the least grain yields were achieved for genotypes Ultan (2,519 kg/ha) and Isfahan1 (1,311 kg/ha), respectively. Grain yield was decreased in some genotypes under 75 % and in all genotypes under 85 % depletion of available water. Based on percentage reduction in grain yield under both 75 and 85 % depletion of soil available water, Isfahan4, Borazjan, Isfahan1, Ahvaz, Ardestan, and Shiraz were recognized as relatively tolerant and Ultan, Shahreza, Kal, and Markazi were identified as relatively sensitive to water stress. The activities of antioxidant enzymes and the contents of carotenoids, proline, and soluble carbohydrates in leaves were increased in most genotypes under stress conditions, and the magnitudes of the increases were greater in the tolerant than in the sensitive genotypes. The results of this experiment showed that the stress-induced increase of antioxidant enzymes and the contents of the compatible solutes in leaves were related to the tolerance of sesame genotypes.     

Possible Involvement of Anti-Oxidant Enzymes in the Cross-Tolerance of the Germination/Growth of Wheat Seeds to Salinity and Heat Stress

The germination/growth of wheat （Triticum aestivum L. cv. Zimai 1） seeds and changes in the activities of superoxide dismutase （SOD）, ascorbate peroxidase （APX）, and catalase （CAT）, as well as in the content of thiobarbituric acid-reactive substances （TBARS）, in response to salt and heat stress, as well as cross-stress, were investigated in the present study. With increasing temperature and decreasing water potential caused by NaCI solution, the germination percentage of seeds and the fresh weight of seedlings decreased markedly, SOD activity increased, activities of APX and CAT decreased distinctly, and the TBARS content increased gradually. Seeds pretreated at 33℃ for different times displayed increased tolerance to subsequent salt stress, enhanced SOD, APX, and CAT activities, and decreased TBARS content. Seeds pretreated at -0.8 MPa NaCI for different times displayed increased tolerance to subsequent heat stress and marked increases in SOD, APX, and CAT activities, which were associated with decreased TBARS content. It is considered that the common component in the cross-tolerance of the germination and growth of wheat seeds to salinity and heat stress is the anti-oxidant enzyme system.     

Salt-Induced Changes in Physio-Biochemical and Antioxidant Defense System in Mustard Genotypes

Salinity stress is a major factor limiting plant growth and productivity of many crops including oilseed. The present study investigated the identification of salt tolerant mustard genotypes and better understanding the mechanism of salinity tolerance. Salt stresses significantly reduced relative water content (RWC), chlorophyll (Chl) content, K⁺ and K⁺ /Na⁺ ratio, photosynthetic rate (P_N), transpiration rate (Tr), stomatal conductance (gs), intercellular CO₂ concentration (Ci) and increased the levels of proline (Pro) and lipid peroxidation (MDA) contents, Na⁺ , superoxide (O₂^•− ) and hydrogen peroxide (H₂O₂) in both tolerant and sensitive mustard genotypes. The tolerant genotypes maintained higher Pro and lower MDA content than the salt sensitive genotypes under stress condition. The activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione peroxidase (GPX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) were increased with increasing salinity in salt tolerant genotypes, BJ-1603, BARI Sarisha-11 and BARI Sarisha-16, but the activities were unchanged in salt sensitive genotype, BARI Sarisha-14. Besides, the increment of ascorbate peroxidase (APX) activity was higher in salt sensitive genotype as compared to tolerant ones. However, the activities of glutathione reductase (GR) and glutathione S-transferase (GST) were increased sharply at stress conditions in tolerant genotypes as compared to sensitive genotype. Higher accumulation of Pro along with improved physiological and biochemical parameters as well as reduced oxidative damage by up-regulation of antioxidant defense system are the mechanisms of salt tolerance in selected mustard genotypes, BJ-1603 and BARI Sarisha-16.