Plant morphological and biochemical responses to field water deficits: I. Responses of glutathione reductase activity and paraquat sensitivity

The effects of water deficits on plant morphology and biochemistry were analyzed in two photoperiodic strains of field-grown cotton (Gossypium hirsutum L.). Plants grown under dryland conditions exhibited a 40 to 85% decrease in leaf number, leaf area index, leaf size, plant height, and total weight per plant. Gross photosynthesis decreased from 0.81 to 0.47 milligram CO₂ fixed per meter per second and the average midday water, osmotic, and turgor potentials decreased to −2.1, −2.4, and 0.3 megapascals, respectively.

There was a progressive increase in glutathione reductase activity and in the cellular antioxidant system in the leaves of stressed plants compared to the irrigated controls. The stress-induced increases in enzyme activity occurred at all canopy positions analyzed.

Irrigation of the dryland plots following severe water stress resulted in a 50% increase in leaf area per gram fresh weight in newly expanded leaves of both strains over the leaves which had expanded under the dryland conditions. Paraquat resistance (a relative measure of the cellular antioxidant system) decreased in the strain T25 following irrigation. Glutathione reductase activities remained elevated in the T25 and T185 leaves which were expanded fully prior to irrigation and in the leaves which expanded following the irrigation treatment.

     

Influence of salt stress on growth,lipid peroxidation and antioxidative enzyme activity in borage (Borago officinalis L.)

Abstract

The effects of increasing salt concentrations on the growth, electrolyte leakage, lipid peroxidation, and major antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) of borage plants were investigated. Plants were grown in half strength of Hoagland nutrient solution added with 0, 25, 50, and 75 mM of NaCl. Most measured parameters were affected by salinity. Increasing salt levels caused a significant reduction in leaf area, stem length, stem diameter, flower number, and dry masses of different organs. Growth of borage plants, in terms of dry weight, was affected. As a consequence of salinity stress, lipid peroxidation and membrane permeability was increased. Antioxidant activity showed an increase in the activity of superoxide dismutase, a non-induced activity of catalase and ascorbate peroxidase, and a slight increase in glutathione reductase activity. The results indicate that borage plants appear to be sensitive to salt stress, since enzymes related to antioxidant enzymatic defense system in treated leaves should be highly active.     

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.     

Delayed wheat flag leaf senescence due to removal of spikelets is associated with increased activities of leaf antioxidant enzymes,reduced glutathione/oxidized glutathione ratio and oxidative damage to mitochondrial proteins

Removal of reproductive ‘sink’ i.e. spikelets from wheat at anthesis delays the rate of flag leaf senescence. In this work, the antioxidant defense was studied in the flag leaf of Triticum aestivum cv. Kalyansona plants showing normal (S + plants) and delayed senescence via removal of spikelets (S? plants). This was done by measurement of metabolites and activities of enzymes such as superoxide dismutase, catalase, guaiacol peroxidase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase. S? plants had higher reduced glutathione/oxidized glutathione (GSH/GSSG) ratio and antioxidant enzyme activities than the control plants and the differences were apparent from 21 days after anthesis (DAA). The removal of the reproductive sink led to an increased antioxidant defense which may be contributing towards the delayed flag leaf senescence in wheat. Chloroplasts and mitochondria, important sources of ROS, were isolated at two stages representing early (7 DAA) and late (21 DAA) senescence. Oxidative damage to proteins was studied in these organelles in relation to SOD and APX. Mitochondria had higher levels of damaged proteins than chloroplasts at 7 DAA in both S+ and S? plants. Higher damage was related to the lower antioxidant enzyme levels of SOD and APX in mitochondria as compared to chloroplasts.     

Both xanthophyll cycle-dependent thermal dissipation and the antioxidant system are up-regulated in grape (Vitis labrusca L cv Concord) leaves in response to N limitation

One-year-old grapevines (Vitis labrusca L. cv. Concord) were supplied with 0, 5, 10, 15, or 20 mM nitrogen (N) in a modified Hoagland's solution twice weekly for 4 weeks. As leaf N decreased in response to N limitation, leaf chlorophyll (Chl) decreased linearly whereas leaf absorptance declined curvilinearly. Compared with high N leaves, low N leaves had lower quantum efficiency of PSII as a result of both an increase in non-photochemical quenching (NPQ) and an increase in closure of PSII reaction centres at midday under high photon flux density (PFD). Both the xanthophyll cycle pool size on a Chl basis and the conversion of violaxanthin (V) to antheraxanthin (A) and zeaxanthin (Z) at noon increased with decreasing leaf N. NPQ was closely related to A+Z expressed either on a Chl basis or as a percentage of the xanthophyll cycle pool. As leaf N increased, superoxide dismutase (SOD) activity on a Chl basis decreased linearly; activities of catalase (CAT) and glutathione reductase (GR) on a Chl basis increased linearly; activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR) and dehydroascorbate reductase (DHAR) expressed on the basis of Chl decreased rapidly first, then gradually reached a low level. In response to N limitation, the contents of ascorbate (AsA), dehydroascorbate (DAsA), reduced glutathione (GSH), and oxidized glutathione (GSSG) increased when expressed on a Chl basis, whereas the ratios of both AsA to DAsA and GSH to GSSG decreased. It is concluded that, in addition to decreasing light absorption by lowering Chl concentration, both xanthophyll cycle-dependent thermal energy dissipation and the antioxidant system are up-regulated to protect low N leaves from photo-oxidative damage under high light.     

Antioxidant enzyme changes in neem,pigeonpea and mulberry leaves in two stages of maturity

Differential expression of antioxidant enzymes in various growth and differentiation stages has been documented in several plant species. We studied here, the difference in the levels of protein content and antioxidant enzymes activity at two stages of maturity, named young and mature in neem (Azadirachta indica A. Juss), pigeonpea (Cajanus cajan (L.) mill sp) and mulberry (Morus Alba L.) leaves. The results showed that detached neem and pigeonpea mature leaves possessed higher activities of catalase (CAT) and peroxidase (POD) and lower activities of polyphenol oxidase (PPO) and ascorbate peroxidase (APX) as compared with young leaves. However, glutathione reductase (GR) showed higher activity in mature leaves of neem, whereas no change in its activity was observed in pigeonpea. On the other hand, antioxidant enzymes in mulberry showed either positive (PPO) or negative (POD, GR, APX) correlation with the progression of leaf maturity. Apparently the trend of changes in antioxidant enzymes activity during leaf development is species-specific: their activity higher at mature stage in some plants and lower in others.     

Time course of antioxidant responses of Capsicum annuum subjected to a progressive magnesium deficiency

The effect of magnesium (Mg²⁺)‐deficiency on the antioxidant responses of Capsicum annuum was investigated over a 60‐day period under controlled conditions. This Mg²⁺‐deficiency aimed to mimic the physiological conditions that plants may experience in the field. At each harvest time, five different leaf‐levels (L2 to L6) were distinguished. L2 and L6 correspond to the second and sixth youngest leaves, respectively. The following parameters were determined: Mg²⁺, chlorophyll and protein contents, total and redox pools of ascorbate and glutathione, and the activities of superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase. Under Mg²⁺‐deficiency, leaf Mg²⁺ contents decreased over time in all leaf‐levels except in the second youngest leaves (L2), where they remained constant at about 0.25% (dry weight basis). Mg²⁺‐deficiency led to an increase in the antioxidant enzyme activities concomitant with an increase in the ascorbate and glutathione pools, whereas total chlorophyll and soluble protein contents decreased. The L2 leaves showed an increase in glutathione reductase activity and in the ascorbate redox state whereas no difference was observed for the other parameters. Superoxide dismutase activities increased in L5 leaves from day 15 and, afterwards, in L3 to L5 leaves, irrespective of Mg²⁺ content. At day 30, glutathione reductase activities increased in L2 to L4 leaves and dehydroascorbate reductase activities in L4 leaves. At day 45, we observed an increase in the ascorbate peroxidase activities in L3 to L5 leaves. At the same time, ascorbate and glutathione pools increased in intermediate leaves, whereas chlorophyll content decreased in L3 and L4 leaves, and protein content decreased in L4 leaves. Results suggest that pepper leaves enhance their defence capacities against oxidative stress by increasing ascorbate more than glutathione synthesis. However, cells showed higher regeneration rates for the glutathione redox state than for the ascorbate redox state.     

Ontogenetic Variation of Nitrogenase, Nitrate Reductase, and Glutamine Synthetase Activities in Oryza sativa

The relationship between the rates of nitrogenase, nitrate reductase, and glutamine synthetase activities, and plant ontogeny in rice (Oryza sativa L.), cultivar `M9', grown in salt marsh sediment with and without nitrate treatment was studied. In both treatments, nitrogenase activity measured as the immediate linear rate of acetylene reduction by bacteria associated with the roots varied with plant age. In control plants, the nitrogenase activity developed during the vegetative stage, peaked during early reproductive growth and then declined. The application of 10 kilograms N per hectare as KNO₃ once every 2 weeks delayed the development of and decreased the nitrogenase activity. The nitrogenase activity in both treatments developed as leaf nitrate reductase activity declined. The per cent nitrogen of roots was negatively correlated with the rates of acetylene reduction during the life cycles of control and nitrate-treated plants. This suggests that the concentration of combined nitrogen in the plants controlled the development and rate of root-associated nitrogenase activity. During reproductive growth, no nitrate reductase activity was detected in the roots from either treatment. In control plants, the patterns of nitrogenase activity and glutamine synthetase activity in the roots were similar. Thus, rice roots have the potential to assimilate ammonia while fixing N₂. During the vegetative and early reproductive stages of growth, the development of maximal rates of nitrogenase activity coincided with an increase of total nitrogen of the plants in both treatments.     

Enhanced sensitivity to oxidative stress in transgenic tobacco plants with decreased glutathione reductase activity leads to a decrease in ascorbate pool and ascorbate redox state

To investigate the possible mechanisms of glutathione reductase (GR) in protecting against oxidative stress, we obtained transgenic tobacco (Nicotiana tabacum) plants with 30–70% decreased GR activity by using a gene encoding tobacco chloroplastic GR for the RNAi construct. We investigated the responses of wild type and transgenic plants to oxidative stress induced by application of methyl viologen in vivo. Analyses of CO₂ assimilation, maximal efficiency of photosystem II photochemistry, leaf bleaching, and oxidative damage to lipids demonstrated that transgenic plants exhibited enhanced sensitivity to oxidative stress. Under oxidative stress, there was a greater decrease in reduced to oxidized glutathione ratio but a greater increase in reduced glutathione in transgenic plants than in wild type plants. In addition, transgenic plants showed a greater decrease in reduced ascorbate and reduced to oxidized ascorbate ratio than wild type plants. However, there were neither differences in the levels of NADP and NADPH and in the total foliar activities of monodehydroascorbate reductase and dehydroascorbate reductase between wild type and transgenic plant. MV treatment induced an increase in the activities of GR, ascorbate peroxidase, superoxide dismutase, and catalase. Furthermore, accumulation of H₂O₂ in chloroplasts was observed in transgenic plants but not in wild type plants. Our results suggest that capacity for regeneration of glutathione by GR plays an important role in protecting against oxidative stress by maintaining ascorbate pool and ascorbate redox state.     

Changes in antioxidant enzymes activity and oxidative stress by abscisic acid and salicylic acid in wheat genotypes

Abscisic acid (ABA) and salicylic acid (SA) were sprayed on leaves of wheat genotypes C 306 and Hira at 25 and 40 d after sowing under moderate water stress (−0.8 MPa) imposed by adding PEG-6000 in nutrient solution. ABA and SA increased the activities of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and catalase in comparison to unsprayed control plants. Both ABA and SA treatments decreased the contents of hydrogen peroxide and thiobarbituric acid reactive substances, a measure of lipid peroxidation, compared to unsprayed plants. The beneficial effect of increase in antioxidant enzymes activity and decrease in oxidative stress was reflected in increase in chlorophyll and carotenoid contents, relative water content, membrane stability index, leaf area and total biomass over control plants. The lower concentrations of ABA (0.5 mM) and SA (1.0 mM) were generally more effective than higher concentrations.     

Differences in water relations and physiological characteristics in leaves of wheat associated with leaf position on the plant

The seasonal change in leaf water potential and its components, stomatal resistance, specific leaf weight, photosynthesis rate, the activities of ribulose-1,5-bisphosphate carboxylase and nitrate reductase, and soluble proteins were measured in flag leaves (ninth from base in position), seventh and fifth leaves of wheat Triticum aestivum L. cv Kalyansona. Flag leaves had a lower water and solute potential and lower or equal turgor pressure than seventh and fifth leaves. These differences were found to be independent of environment. The rate of photosynthesis and nitrate reductase activity were always lower in fifth and seventh leaves than in flag leaf. The photosynthetic efficiency in flag leaves appeared to be associated with lower stomatal resistance and higher specific leaf weight. The relations between leaf water potential and relative water content showed a change with leaf position. This change possibly allows flag leaf to maintain its functional efficiency despite its lower water potential.     

Carbohydrate Metabolism in Leaf Meristems of Tall Fescue : II. Relationship to Leaf Elongation Rates Modified by Nitrogen Fertilization

Our objective was to examine alterations in carbohydrate status of leaf meristems that are associated with nitrogen-induced changes in leaf elongation rates of tall fescue (Festuca arundinacea Schreb.). Dark respiration rates, concentrations of nonstructural carbohydrates, and soluble proteins were measured in leaf intercalary meristems and adjacent segments of elongating leaves. The two genotypes used differed by 43% in leaf elongation rate. Application of high nitrogen (336 kilograms per hectare) resulted in 140% higher leaf elongation rate when compared to plants receiving low nitrogen (22 kilograms per hectare). Leaf meristems of plants receiving high and low nitrogen had dark respiration rates of 5.4 and 2.9 microliters O₂ consumed per milligram structural dry weight per hour, respectively. Concentrations of soluble proteins were lower while concentrations of fructan tended to be slightly higher in leaf meristems of low-nitrogen plants when compared to high-nitrogen plants. Concentrations of reducing sugars, nonreducing sugars, and takadiastase-soluble carbohydrate of leaf meristems were not affected by nitrogen treatment. Total nonstructural carbohydrates of leaf meristems averaged 44 and 39% of dry weight for low- and high-nitrogen plants, respectively. Within the leaf meristem, approximately 74 and 34% of the pool of total nonstructural carbohydrate could be consumed per day in high- and low-nitrogen plants, respectively, assuming no carbohydrate import to the meristem occurred. Plants were able to maintain high concentrations of nonstructural carbohydrates in leaf meristems despite a 3-fold range in leaf elongation rates, suggesting that carbohydrate synthesis and transport to leaf intercalary meristems may not limit leaf growth of these genotypes.     

Responses of superoxide dismutase and glutathione reductase activities in cotton leaf tissue exposed to an atmosphere enriched in oxygen

Responses of superoxide dismutase (EC 1.15.1.1) and glutathione reductase (EC 1.6.4.2) activities were evaluated in leaf tissue from intact cotton plants (Cotton Branch 1697) which were exposed to 75% O₂, 350 microliters per liter CO₂ for 48 hours. Soluble protein was extracted from O₂-treated and control tissue, and enzyme levels were determined. Superoxide dismutase activity in cotton leaf tissue was high (26 units per milligram protein) under normal conditions of 21% O₂, saturating light, and limiting CO₂, and neither qualitative nor quantitative differences in the cyanide-sensitive or -insensitive forms of the enzyme occurred in response to hyperoxic conditions. Glutathione reductase activity, however, was 2- to 3-fold higher in extracts from tissue exposed to 75% O₂. No increase in activity was observed for the peroxisomal enzymes, glycolate oxidase (EC 1.1.3.1) and catalase (EC 1.11.1.6). Results are consistent with an integrated pathway involving superoxide dismutase and glutathione reductase for protection of sensitive leaf components against detrimental effects of intermediate reduction products of O₂.     

Developmental changes of antioxidative systems in tobacco leaves as affected by limited sucrose export in transgenic plants expressing yeast-invertase in the apoplastic space

It is generally believed that a restricted export of carbohydrates from source leaves causes oxidative stress because of an enhanced utilisation of O₂ instead of NADP⁺ as electron acceptor in photosynthesis. To test this hypothesis, developmental changes of antioxidative systems were investigated in wild-type and transgenic tobacco (Nicotiana tabacum L.) suffering from disturbed sink-source relations by expression of yeast invertase in the apoplastic space. Young expanding leaves of the wild type contained higher activities of Superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6), dehydroascorbate reductase (EC 1.8.5.1), glutathione reductase (EC 1.6.4.2) and a higher glutathione content than mature source leaves. The activity of monodehydroascorbate-radical reductase (EC 1.1.5.4) and the ascorbate content remained unaffected by the developmental stage in the wild type. In young expanding leaves of the transgenic plants the capacity of the antioxidative systems was similar to or higher than in corresponding leaves from the wild type. Source leaves of transgenic tobacco with an increased carbohydrate content showed a small chlorophyll loss, an increased malondialdehyde content, a selective loss of the activities of Cu/Zn-superoxide dismutase isoenzymes and a fourfold decrease in ascorbate compared with the wild type. There was no evidence that the protection from H₂O₂ was insufficient since source leaves of transgenic tobacco contained increased activities of catalase, ascorbate peroxidase, and monodehydroascorbate-radical reductase and an increased ascorbate-to-dehydroascorbate ratio compared with source leaves of the wild type. In severely chlorotic leaf sections of the transgenic plants, most components of the antioxidative system were lower than in green leaf sections, but the ascorbate-to-dehydroascorbate ratio was increased. These results suggest that carbohydrate-accumulating cells have an increased availability of reductant, which can increase the degree of reduction of the ascorbate system via glutathione-related systems or via the activity of monodehydroascorbate-radical reductase. At the same time, transgenic tobacco leaves seem to suffer from an increased oxidative stress, presumably as a result of a decreased consumption of O ₂ ^.- by Cu/Zn-superoxide dismutases in the chloroplasts. There was no evidence that carbohydrate-accumulating leaves acclimated to enhanced O ₂ ^.- production rates in the chloroplasts.     

Salicylic acid pretreatment induces drought tolerance and delays leaf rolling by inducing antioxidant systems in maize genotypes

Salicylic acid (SA) acts as an endogenous signal molecule responsible for inducing abiotic stress tolerance in plants. In this study, the role of SA in improving drought tolerance in two maize cultivars (Zea mays L.) differing in their tolerance to drought was evaluated. The plants were regularly watered per pot and grown until the grain filling stage (R2) under a rainout shelter. At stage R2, parts of the plants were treated with SA, after which drought stress was applied. Leaf samples were harvested on the 10th and 17th days of the drought. Some antioxidant enzyme activity, such as the superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) content, was measured during the drought period. Exogenous SA prevented water loss and delayed leaf rolling in comparison with control leaves in both cultivars. As a consequence of drought stress, lipid peroxidation, measured in terms of malondialdehyde content, was prevented by SA. SA pretreatment induced all antioxidant enzyme activities, and to a greater extent than the control leaves, during drought. SA also caused a reduction in the ascorbate (ASC) and glutathione (GSH) content in two maize cultivars. The H₂O₂ level was higher in SA pretreated plants than the controls in both cultivars. Pretreatment with SA further enhanced the activities of antioxidant enzymes and the concentrations of non-enzymatic antioxidants in the tolerant cultivar compared with the sensitive cultivar. Results suggested that exogenous SA could help reduce the adverse effects of drought stress and might have a key role in providing tolerance to stress by decreasing water loss and inducing the antioxidant system in plants with leaf rolling, an alternative drought protection mechanism.     

Role of salicylic acid in regulating ultraviolet radiation-induced oxidative stress in pepper leaves

The effect of salicylic acid (SA) counteracting the UV-A, UV-B, and UV-C-induced action on pepper (Capsicum annuum L.) plants was studied. For this purpose, the activities of antioxidant enzymes (peroxidase, polyphenol oxidase, ascorbate peroxidase, catalase, and glutathione reductase) were measured. Plants were sprayed with SA and treated with UV-A (320–390 nm), UV-B (312 nm), and UV-C (254 nm) radiation with a density of 6.1, 5.8, and 5.7 W/m². The activities of antioxidant enzymes were enhanced in leaves in response to UV-B and UV-C radiation. SA treatment moderated an increase in the activities of some antioxidant enzymes (peroxidase, ascorbate peroxidase, catalase, and glutathione reductase) in plants that were treated with UV radiation. The activity of antioxidant enzyme polyphenol oxidase in plants that were treated with UV-B, UV-C, and SA was significantly increased. The aim of the present study was to investigate the possible protective effect of SA treatment on UV-A, UV-B, and UV-C stress.     

Glutathione-Mediated Alleviation of Chromium Toxicity in Rice Plants

A hydroponic experiment was conducted to determine the possible effect of exogenous glutathione (GSH) in alleviating chromium (Cr) stress through examining plant growth, chlorophyll contents, antioxidant enzyme activity, and lipid peroxidation in rice seedlings exposed to Cr toxicity. The results showed that plant growth and chlorophyll content were dramatically reduced when rice plants were exposed to 100 μM Cr. Addition of GSH in the culture solution obviously alleviated the reduction of plant growth and chlorophyll content. The activities of some antioxidant enzymes, including superoxide dismutase, catalase (CAT) and glutathione reductase in leaves, and CAT and glutathione peroxidase in roots showed obvious increase under Cr stress. Addition of GSH reduced malondialdehyde accumulation and increased the activities of these antioxidant enzymes in both leaves and roots, suggesting that GSH may enhance antioxidant capacity in Cr-stressed plants. Furthermore, exogenous GSH caused significant decrease of Cr uptake and root-to-shoot transport in the Cr-stressed rice plants. It can be assumed that GSH is involved in Cr compartmentalization in root cells.     

Activation of ascorbate-glutathione cycle inArabidopsis leaves in response to aminotriazole

Aminotriazole(AT)-induced changes in growth, hydrogen peroxide content and activities of H₂O₂-scavenging antioxidant enzymes were investigated in the growing leaves ofArabidopsis plants (Arabidopsis thaliana cv Columbia). Catalase activity of rosette leaves was reduced by 65% with an application of 0.1 mM AT (a herbicide known as a catalase inhibitor), whereas the leaf growth and H₂O₂ content were almost unaffected. However, an approximate 1.6 to 2-fold increase in cytosolic ascorbate peroxidase (APX) activity concomitant with a substantial activation of glutathione reductase (GR) (approx. 22% increase) was observed during leaf growth in the presence of 0.1 mM AT. The activity of cytosolic APX in leaves was also increased by 1.8-fold with an application of exogenous 2 mM paraquat (an inducer of H₂O₂ production in plant cells) in the absence of AT. These results collectively suggest that (a) cytosolic APX and GR operate to activate an ascorbate-glutathione cycle for the removal of H₂O₂ under severe catalase deactivation, and (b) the expression of APX seems to be regulated by a change of the endogenous H₂O₂ level in leaf cells.     

Hydrogen peroxide pre-treatment induces salt-stress acclimation in maize plants

The effect of exogenously applied H2O2 on salt stress acclimation was studied with regard to plant growth, lipid peroxidation, and activity of antioxidative enzymes in leaves and roots of a salt-sensitive maize genotype. Pre-treatment by addition of 1 microM H2O2 to the hydroponic solution for 2 days induced an increase in salt tolerance during subsequent exposure to salt stress. This was evidenced by plant growth, lipid peroxidation and antioxidative enzymes measurements. In both leaves and roots the variations in lipid peroxidation and antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase, and catalase) activities of both acclimated and unacclimated plants, suggest that differences in the antioxidative enzyme activities may, at least in part, explain the increased tolerance of acclimated plants to salt stress, and that H2O2 metabolism is involved as signal in the processes of maize salt acclimation.     

Manipulation of Catalase Levels Produces Altered Photosynthesis in Transgenic Tobacco Plants

Constructs containing the cDNAs encoding the primary leaf catalase in Nicotiana or subunit 1 of cottonseed (Gossypium hirsutum) catalase were introduced in the sense and antisense orientation into the Nicotiana tabacum genome. The N. tabacum leaf cDNA specifically overexpressed CAT-1, the high catalytic form, activity. Antisense constructs reduced leaf catalase specific activities from 0.20 to 0.75 times those of wild type (WT), and overexpression constructs increased catalase specific activities from 1.25 to more than 2.0 times those of WT. The NADH-hydroxypyruvate reductase specific activity in transgenic plants was similar to that in WT. The effect of antisense constructs on photorespiration was studied in transgenic plants by measuring the CO₂ compensation point (Γ) at a leaf temperature of 38°C. A significant linear increase was observed in Γ with decreasing catalase (at 50% lower catalase activity Γ increased 39%). There was a significant temperature-dependent linear decrease in Γ in transgenic leaves with elevated catalase compared with WT leaves (at 50% higher catalase Γ decreased 17%). At 29°C, Γ also decreased with increasing catalase in transgenic leaves compared with WT leaves, but the trend was not statistically significant. Rates of dark respiration were the same in WT and transgenic leaves. Thus, photorespiratory losses of CO₂ were significantly reduced with increasing catalase activities at 38°C, indicating that the stoichiometry of photorespiratory CO₂ formation per glycolate oxidized normally increases at higher temperatures because of enhanced peroxidation.