Antioxidative defense system in pigeonpea roots under waterlogging stress

Pigeonpea [Cajanus cajan (L.) Millsp.] is a waterlogging-sensitive legume crop. We studied the effect of waterlogging stress on hydrogen peroxide (H₂O₂) content, lipid peroxidation and antioxidant enzyme activities in two pigeonpea genotypes viz., ICPL-84023 (waterlogging resistant) and MAL-18 (waterlogging susceptible). In a pot experiment, waterlogging stress was imposed for 6 days at early vegetative stage (20 days after sowing). Waterlogging treatment significantly increased hydrogen peroxide accumulation and lipid peroxidation, which indicated the extent of oxidative injury posed by stress conditions. Enzyme activities of peroxidase (POX), catalase (CAT), ascorbate peroxidase (APX), superoxide dismutase (SOD) and polyphenol oxidase (PPO) increased in pigeonpea roots as a consequence of waterlogged conditions, and all the enzyme activities were significantly higher in waterlogged ICPL-84023 than in MAL-18. POX activity was the maximum immediately after imposing stress, therefore, it was suggested to be involved in early scavenging of H₂O₂, while rest of the enzymes (CAT, APX, SOD and PPO) were more important in late responses to waterlogging. Present study revealed that H₂O₂ content is directly related to lipid peroxidation leading to oxidative damage during waterlogging in pigeonpea. Higher antioxidant potential in ICPL-84023 as evidenced by enhanced POX, CAT, APX, SOD and PPO activities increased capacity for reactive oxygen species (ROS) scavenging and indicated relationship between waterlogging resistance and antioxidant defense system in pigeonpea.     

Effect of Arbuscular Mycorrhizal Inoculation on Salt-induced Nodule Senescence in <Emphasis Type="Italic">Cajanus cajan</Emphasis> (Pigeonpea)

Many physiological and biochemical plant processes affected by salt stress trigger premature nodule senescence and decrease their ability to fix nitrogen. The objective of this study was to evaluate the role of arbuscular mycorrhiza (AM) in moderating salt-induced premature nodule senescence in Cajanus cajan (L.) Millsp. Greenhouse experiments were conducted in which the plants were exposed to salinity stress of 4, 6, and 8 dSm⁻¹. Various parameters linked to nodule senescence were assessed at 80 days after sowing. Nodulation, leghemoglobin content, and nitrogenase enzyme activity measured as acetylene-reducing activity (ARA) were evaluated. Two groups of antioxidant enzymes were studied: (1) enzymes involved in the detoxification of O₂⁻ radicals and H₂O₂, namely, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX), and (2) enzymes that are important components of the ascorbate glutathione pathway responsible for the removal of H₂O₂, namely, glutathione reductase (GR) and ascorbate peroxidase (APOX). Exposure of plants to salinity stress enhanced nodule formation; however, nodule growth suffered remarkably and a marked decline in nodule biomass, relative permeability, and lipid peroxidation was observed. Leghemoglobin content and ARA were reduced under saline conditions. AM significantly improved nodulation, leghemoglobin content, and nitrogenase activity under salt stress. Activities of SOD, CAT, APOX, POX, and GR increased markedly in mycorrhizal-stressed plants. A synthesis of the evidence obtained in this study suggests a correlation between enhanced levels of antioxidant enzyme activities, reduced membrane permeability, reduced lipid peroxidation, and improved nitrogen-fixing efficiency of AM plants under stressed and unstressed conditions. These factors could be responsible for the protective effects of mycorrhiza against stress-induced premature nodule senescence.     

Sphaerophysa kotschyana, an endemic species from Central Anatolia: antioxidant system responses under salt stress

Sphaerophysa kotschyana is a Turkish endemic and endangered plant that grows near Salt Lake, in Konya, Turkey. However, little is known about the ability of this plant to generate/remove reactive oxygen species (ROS) or its adaptive biochemical responses to saline environments. After exposure of S. kotschyana to 0, 150, and 300 mM NaCl for 7 and 14 days, we investigated (1) the activities and isozyme compositions of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), and glutathione reductase (GR); (2) the oxidative stress parameters NADPH oxidase (NOX) activity, lipid peroxidation (MDA), total ascorbate (tAsA) content, and total glutathione content (tGlut); and (3) ROS levels for superoxide anion radical (O ₂ ^·? ), hydrogen peroxide (H₂O₂), hydroxyl radicals (OH·), and histochemical staining of O ₂ ^·? and H₂O₂. H₂O₂ content increased after 14 days of salt stress, which was consistent with the results from histochemical staining and NOX activity measurements. In contrast, oxidative stress induced by 150 mM NaCl was more efficiently prevented, as indicated by low malondialdehyde (MDA) levels and especially at 7 days, by increased levels of SOD, POX, APX, and GR. However, at 300 mM NaCl, decreased levels of protective enzymes such as SOD, CAT, POX, and GR, particularly with long-term stress (14 days), resulted in limited ROS scavenging activity and increased MDA levels. Moreover, at 300 mM NaCl, the high H₂O₂ content caused oxidative damage rather than inducing protective responses against H₂O₂. These results suggest that S. kotschyana is potentially tolerant to salt-induced damage only at low salt concentrations.     

Some Enzymes of Hydrogen Peroxide Metabolism in Leaves and Root Nodules of Medicago sativa

Leaves and nodules (bacteroids and cytosol) of alfalfa (Medicago sativa L. cv Aragon) plants inoculated with Rhizobium meliloti strain 102F51 have been analyzed for the presence of the enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (EC 1.11.1.6), and peroxidase (EC 1.11.1.7). All three fractions investigated (leaves, bacteroids, and nodular cytosol) show Cu,Zn-SOD activity. Besides, the bacteroids and cytosol of nodules possess CN⁻-insensitive SOD activities. Studies of SOD inactivation with H₂O₂ indicate that, very likely, a Mn-SOD is present in the bacteroids, and suggest that the cytosol contain both Mn-SOD and Fe-SOD. Bacteroids show high catalase activity but lack peroxidase. By contrast, the nodule cytosol exhibits an elevated peroxidase activity as compared with the foliar tissue; this activity was completely inhibited by 50 to 100 micromolar KCN. The significantly lower contents of H₂O₂ and malondialdehyde (a product of lipid peroxidation) in nodules with respect to those in leaves reveal that the above-mentioned bacteroid and cytosol enzymes act in an efficient and combined manner to preserve integrity of nodule cell membranes and to keep leghemoglobin active.     

Nodule Senescence in Medicago truncatula�CSinorhizobium Symbiosis Under Abiotic Constraints: Biochemical and Structural Processes Involved in Maintaining Nitrogen-Fixing Capacity

Nitrogen-fixing capacity (NFC) in nodules of four Medicago truncatula lines inoculated with four strains of Sinorhizobium was assessed, during the plant life cycle, in relation to parameters identified as indices of plant growth, photosynthetic capacity, nodule integrity, and functioning. Differences in duration of the NFC period were observed among symbiotic associations and were correlated with variability on plant biomass production. Senescence appearance and vigor varied in parallel with structural, physiological, and biochemical stability of nodules. Maintenance of a longer high-NFC period was correlated to a higher stimulation of antioxidant enzymes, mainly superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (POX, EC 1.11.1.7), and a consequent longer maintenance of membrane integrity and nodule structure within the first stages of senescence. Salinity and drought stresses interfered with nodule functioning and triggered fast and global nodule senescence, albeit a superiority of nodules having a long high-NFC period. The protective role of POX activity on salt- and drought-stressed nodules was revealed. On the other hand, SOD stimulation was independent of stress application. Another strategy allowing the maintenance of longer NFC in salt-stressed nodules could be the accumulation of starch granules in the senescence-functioning interface of nodules. This finding is currently under investigation. Interestingly, the symbioses with different behaviors of nodule senescence identified in this work would be useful bases for biochemical, genomic, and proteomic studies dissecting nodule senescence.     

Antioxidant Enzyme Activities during in vitro Morphogenesis of Gladiolus and the Effect of Application of Antioxidants on Plant Regeneration

Activity of antioxidant enzymes was evaluated during somatic embryogenesis and shoot organogenesis from cultured leaf segments of Gladiolus hybridus Hort. The effect of exogenous antioxidants on somatic embryogenesis and shoot organogenesis has also been monitored. Activity of superoxide dismutase (SOD) gradually increased during somatic embryogenesis. while activities of catalase (CAT) and peroxidase (POX) decreased. In contrast, increase in CAT and POX activity and a concomitant decrease in SOD activity were noted during shoot organogenesis. Exogenous application of antioxidants such as glutathione (GSH), α-tocopherol and ascorbate (AA) inhibited somatic embryogenesis but stimulated shoot organogenesis. The frequency of somatic embryogenesis increased with the addition of H₂O₂. However, H₂O₂ inhibited shoot organogenesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Salicylic acid increases tolerance to oxidative stress induced by hydrogen peroxide accumulation in leaves of cadmium-exposed flax (Linum usitatissimum L.)

The aim of this study is to investigate the impacts of exogenous salicylic acid (SA) pretreatments on hydrogen peroxide (H₂O₂) accumulation, protein oxidation, and H₂O₂-scavenging enzymes in leaves of Cd-treated flax seedlings. Cd-enhanced H₂O₂ levels were related to increased activities of guaiacol peroxidase (POX, EC 1.11.1.7) and ascorbate peroxidase (APX, EC 1.11.1.11), and were independent of changes in catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) activities. In control flax seedlings, exogenous SA pretreatments inhibited the activity of CAT, resulted in an enhanced production of H₂O₂ suggesting that SA requires H₂O₂ to initiate an oxidative stress. However, although leaves of Cd-free flax seedlings pretreated with SA accumulated in vivo H₂O₂ by 1.2-fold compared with leaves of Cd-only exposed ones; the damage to growth and proteins after the exposure to Cd was significantly less, indicating that SA can regulate the Cd-induced oxidative stress. Moreover, the Cd-treated seedlings primed with SA exhibited a higher level of total antioxidant capacities and increased activities of H₂O₂-detoxifying enzymes.     

Changes in activities of antioxidant enzymes and their gene expression during leaf development of sweetpotato

To understand the functions of antioxidant enzymes during leaf development in sweetpotato, we investigated the activities of several antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), ascorbate peroxidase (APX) and catalase (CAT). Significant increases were observed in the activities of SOD, POX and APX during the late stage of leaf development, whereas CAT activity increased during the early developmental stage. By RT-PCR analysis, various POX and APX genes showed differential expression patterns during leaf development. Four POX genes swpa3, swpa4, swpa6, swpb4 and one APX gene swAPX1 exhibited high levels of gene expression during the senescence stage of leaf development, but two POX genes, swpa1 and swpa7 were preferentially expressed at both the mature green and the late senescence stages of leaf development. These results indicate that hydrogen peroxide (H₂O₂)-related antioxidant enzymes are differentially regulated in the process of leaf development of sweetpotato.     

Exogenous application of penconazole regulates plant growth and antioxidative responses in salt-stressed Mentha pulegium L.

The mechanism of growth amelioration in salt-stressed pennyroyal (Mentha pulegium L.) was investigated by exogenous application of penconazole (PEN). Seven weeks after sowing, seedlings were treated with increasing NaCl concentrations (0, 25, 50, and 75 mM) with or without PEN (15 mg l^?1) and were harvested randomly at different times. Results showed that some growth parameters and the relative water content (RWC) decreased under salt stress, while lipid peroxidation, H₂O₂ content, activities of superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POX; EC 1.11.1.7), polyphenol oxidase (PPO; EC 1.10.3.1), catalase (CAT; EC 1.11.1.6), and ascorbate peroxidase (APX; EC 1.11.1.1) remarkably increased. Exogenous application of PEN increased some growth parameters, RWC, antioxidant enzyme activities, and H₂O₂ content, but the effects of PEN were more significant under salt stress conditions. PEN treatment also decreased lipid peroxidation. These results suggest that PEN-induced tolerance to salt stress in M. pulegium plants may be related to regulation of antioxidative responses and H₂O₂ level.     

Free radical formation and activity of antioxidant enzymes in lupin roots exposed to lead

Inhibition of root growth and modification of root morphology are the most sensitive responses of Lupinus luteus cv. Ventus L. to lead ions - Pb(NO₃)₂. Using electron paramagnetic resonance (EPR), we found that at the lead concentration of 150 mg.L^–1, the level of free radicals remained at control level, whereas at the higher, sublethal concentration of 350 mg.L^–1, they markedly increased. The EPR signal with the g-value at the maximum absorption of 2.0053 implied that the paramagnetic radical is derived from a quinone. The response of antioxidant enzymes, such as superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POX, EC 1.11.1.7) and ascorbate peroxidase (APOX, EC 1.11.1.11), to exogenously applied lead ions was also examined. Enzyme activity was estimated as a function of time and concentration. Native polyacrylamide gel electrophoresis followed by specific staining revealed an increase in the activity of SOD, CAT, POX and APOX coinciding with the time of cultivation. A lead-dependent increase in activities of SOD and POX from root tip extracts was observed, whereas CAT and APOX activities decreased at the higher lead concentrations. These results suggest that at higher lead concentrations, the formation of both free radicals and reactive oxygen species is beyond the capacity of the antioxidant system, which in turn may contribute to the reduced root growth.     

UV-B induced stress responses in three rice cultivars

UV-B responses of three rice (Oryza sativa L.) cultivars (Sasanishiki, Norin 1 and Surjamkhi) with different photolyase activity were investigated. Carbon dioxide assimilation data support that Sasanishiki was less sensitive to UV-B than Norin 1 and Surjamkhi. UV-B radiation sharply decreased the content of Rubisco protein in Surjamkhi and has no effect in Sasanishiki. The photochemical activities of photosystem (PS) 1 and PS 2 was slightly affected by UV-B treatment. The content of H₂O₂ and the activities of antioxidant enzymes, catalase (CAT), peroxides (POX) and superoxide dismutase (SOD) were enhanced after UV-B treatment. The activities of CAT and POX isoenzymes in Sasanishiki were more enhanced by UV-B radiation than those in Norin 1 and Surjamkhi.     

Pre-treatment with H<Subscript>2</Subscript>O<Subscript>2</Subscript> induces salt tolerance in Barley seedlings

Barley seedlings were pre-treated with 1 and 5 μM H₂O₂ for 2 d and then supplied with water or 150 mM NaCl for 4 and 7 d. Exogenous H₂O₂ alone had no effect on the proline, malondialdehyde (MDA) and H₂O₂ contents, decreased catalase (CAT) activity and had no effect on peroxidase (POX) activity. Three new superoxide dismutase (SOD) isoenzymes appeared in the leaves as a result of 1 μM H₂O₂ treatment. NaCl enhanced CAT and POX activity. SOD activity and isoenzyme patterns were changed due to H₂O₂ pre-treatment, NaCl stress and leaf ageing. In pre-treated seedlings the rate of ¹⁴CO₂ fixation was higher and MDA, H₂O₂ and proline contents were lower in comparison to the seedlings subjected directly to NaCl stress. Cl⁻ content in the leaves 4 and 7 d after NaCl supply increased considerably, but less in pre-treated plants. It was suggested that H₂O₂ metabolism is involved as a signal in the processes of barley salt tolerance.     

ANTIOXIDANT ENZYME RESPONSE AND REACTIVE OXYGEN SPECIES PRODUCTION IN MARINE RAPHIDOPHYTES1

Raphidophytes (class Raphidophyceae) produce high levels of reactive oxygen species (ROS), yet little is known regarding cellular scavenging mechanisms needed for protection against these radicals. Enzymatic activities of the antioxidants superoxide dismutase (SOD) and catalase (CAT) were measured in conjunction with the production of superoxide (O₂^??) and hydrogen peroxide (H₂O₂) in batch cultures of five different raphidophytes species during early exponential, late‐exponential, and stationary growth phases. The greatest concentrations of O₂^?? per cell were detected during exponential growth with reduced levels in stationary phases in raphidophytes Heterosigma akashiwo (Hada) Hada ex Y. Hara et Chihara, Chattonella marina (Subrahman.) Y. Hara et Chihara, and Chattonella antiqua (Hada) Ono (strain 18). Decreasing trends from exponential to stationary phases for SOD activity and H₂O₂ per cell were observed in all species tested. Significant correlations between O₂^?? per cell and SOD activity per cell over growth phase were only observed in three raphidophytes (Heterosigma akashiwo, Chattonella marina, and Chattonella antiqua strain 18), likely due to different cellular locations of externally released O₂^?? radicals and intracellular SOD enzymes measured in this study. CAT activity was greatest at early exponential phase for several raphidophytes, but correlations between H₂O₂ per cell and CAT activity per cell were only observed for Fibrocapsa japonica Toriumi et Takano, Chattonella antiqua (strain 18), and Chattonella subsalsa Biecheler. Our results suggest that SOD and CAT play important protective roles against ROS during exponential growth of several raphidophytes, while other antioxidant pathways may play a larger role for scavenging ROS during later growth.     

Antioxidant activity and ROS tolerance in triticale (×Triticosecale Wittm.) anthers affect the efficiency of microspore embryogenesis

To verify the hypothesis that cell redox status regulates the process of microspore embryogenesis (ME), reactive oxygen species (ROS) generation and the activity of enzymatic and non-enzymatic antioxidants were analyzed in eight doubled haploid lines of triticale with significant differences in embryogenic potential. The analyses were performed in anthers excised from freshly cut tillers (control) and from low temperature (LT) pre-treated tillers (3 weeks at 4 °C) in which ME has been initiated. Significant associations between ME effectiveness and the variables studied were found. In control cultures, high superoxide dismutase (SOD) activity appeared crucial for microspore viability. On the other hand, positive though non-linear correlation between ME effectiveness and H₂O₂ generation, and negative correlation with catalase (CAT) activity suggest that some threshold level of H₂O₂ is important for successful ME initiation. LT tillers pre-treatment significantly increased H₂O₂ accumulation, which had a negative effect on ME effectiveness. However, even high level of H₂O₂ did not endanger cell viability as long as the cells exhibited high activity of ROS-decomposing enzymes (SOD, CAT and POX). The ability to sustain antioxidative enzyme activity under cold stress in the dark was another important requirement for high effectiveness of ME, allowing for the generation of the signal initiating microspore reprogramming and simultaneously protecting the cells from the toxic effects of ROS production. The role of antioxidative enzymes cannot be replaced even by high activity of non-enzymatic antioxidants. In conclusion, genetically controlled but environmentally modified cell tolerance to oxidative stress seems to play an important role in triticale ME.     

Differential responses of antioxidant enzymes to aluminum toxicity in two rice (Oryza sativa L.) cultivars with marked presence and elevated activity of Fe SOD and enhanced activities of Mn SOD and catalase in aluminum tolerant cultivar

Seedlings of two Indica rice (Oryza sativa L.) cvs. HUR-105 and Vandana, differing in Al-tolerance were used to identify the key mechanisms involved in their differential behaviour towards Al toxicity. Cv. HUR-105 appeared to be Al sensitive by showing significant reduction (p ≤ 0.01) in root/shoot length, fresh weight, dry weight and water content in presence of 421 μM Al³⁺ in growth medium whereas cv. Vandana appeared to be fairly Al³⁺ tolerant. A conspicuous and significant reduction in dry weight of root and shoot was observed in Al sensitive cv. HUR-105 with 178 μM Al³⁺ treatment for 3 days. Al was readily taken up by the roots and transported to shoots in both the rice cultivars. Localization of absorbed Al was always greater in roots than in shoots. Our results of the production of reactive oxygen species (ROS) H₂O₂ and O₂ ^.? and activities of major antioxidant enzymes such as total superoxide dismutase (SOD), Cu/Zn SOD, Mn SOD, Fe SOD, catalase (CAT) and guaiacol peroxidase revealed Al induced higher oxidative stress, greater production of ROS and lesser capacity to scavenge ROS in cv. HUR-105 than Vandana. With Al treatment, higher oxidative stress was noted in shoots than in roots. Greatly enhanced activities of SOD (especially Fe and Mn SOD) and CAT in Al treated seedlings of cv. Vandana suggest the role of these enzymes in Al tolerance. Furthermore, a marked presence of Fe SOD in roots and shoots of the seedlings of Al tolerant cv. Vandana and its significant (p ≤ 0.01) increase in activity due to Al-treatment, appears to be the unique feature of this cultivar and indicates a vital role of Fe SOD in Al-tolerance in rice.     

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.     

Short-term effect of nitrate or water stress on nitrate reduction and malate fermentation pathways in yellow lupine (<Emphasis Type="Italic">Lupinus luteus</Emphasis>) nodules

Seven-week-old plants (symbiotic stage) of yellow lupine (Lupinus luteus L. cv. Ventus) were subjected for 8 days to 5 mM nitrate treatment or to drought stress to search for possible activation of bacteroidal nitrate and nitrite reductases. Both treatments affected activities of malate dehydrogenase and aspartate aminotransferase in nodule cytosol and therefore are presumed to impose O₂-limitation to nodule metabolism. However, no significant symptoms of senescence of nodules were found. Both nitrate treatment and drought stress increased rhizobial nitrate and nitrite reductase activities in contrast to noted decrease of corresponding activities in nodule cytoplasm. Differential regulation supports the hypothesis that bacteroidal enzymes can act in dissimilatory mode when nodule respiration is limited due to environmental stresses.     

Reactive Oxygen Species and Relative Enzyme Activities in the Development of Aerial Roots of Chinese Banyan (Ficus microcarpa)

Although previous research has indicated that reactive oxygen species (ROS) regulate cell extension and tissue ontogenesis, the functions of ROS in aerial roots have not been previously studied. This research evaluated ROS production and dissipation in aerial roots of Chinese banyan (Ficus microcarpa). Aerial root segments (4 cm long) were cut from trees and divided into developmental zones 1, 2, and 3 (0–5, 5–15, and 15–25 mm from root tip, respectively). According to histochemical and biochemical determinations, production of the superoxide radical (O ₂ ^·– ), hydrogen peroxide (H₂O₂), and the hydroxyl radical (^·OH) decreased from zone 1 to zone 3. The detected ROS increased with the application of exogenous stimulators of ROS generation and decreased with the application of exogenous inhibitors of ROS generation. Based on protein content, superoxide dismutase (SOD) activity increased but peroxidase (POD) and catalase (CAT) activities decreased from zone 1 to zone 3, whereas based on root segment fresh weight, SOD and CAT activities did not differ among the zones but POD activity decreased from zone 1 to 3. We conclude that ROS are generated mainly in the rapidly developing zones of aerial roots and suggest that NADPH oxidase, POD, and SOD control ROS generation. POD activity and the hydroxyl cycle seem particularly important in ROS generation in aerial roots.     

Study of antioxidant enzymes activity and isozymes pattern in hairy roots and regenerated plants in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis>

Hairy root disease is caused by the infection of wounded higher plants with Agrobacterium rhizogenes. Transformation of tissues or plants with A. rhizogenes, and with rol genes, as well as hairy roots may produce alterations in the plant secondary metabolism. H₂O₂ and other ROS are involved as a signal in secondary metabolite production pathway and play a key role in plant defensive reactions. In this work, the effect of A. rhizogenes T-DNA on nicotine content, antioxidant enzymes activity, H₂O₂ production, pattern of peroxidase (POX) and superoxide dismutase (SOD) isozymes in hairy roots and regenerated plants were studied. Rise in SOD and POX activities in the transformed lines of TRa and TRb and in the resultant regenerated plants, also the decreased level of H₂O₂ in them, compared with the untransformed lines indicates that, the T-DNA genes expression of A. rhizogenes probably decreases H₂O₂ level by increasing the production of antioxidant enzymes. Decrees the level of H₂O₂ content in TRc line in spite of the similarity of antioxidant enzyme activity in comparison with normal root, indicate that A. rhizogenes activate other mechanisms except SOD and POX enzyme for reducing H₂O₂ level.     

Antioxidant enzyme activities and isozyme pattern in hairy roots and regenerated tobacco plants

Hairy root disease is caused by infection of wounded higher plants with Agrobacterium rhizogenes. Transformation of tissues or plants with A. rhizogenes, as well as transformation with rol genes, in addition to hairy roots, may produce alterations in the plant secondary metabolism. H₂O₂ and other ROS are involved as signals in secondary metabolite production pathways and play a key role in plant defense reactions. In this work the effects of A. rhizogenes rol genes on nicotine content, antioxidant enzymes activity, H₂O₂ production, the pattern of peroxidase (POX) and superoxide dismutase (SOD) isozymes in hairy roots and regenerated Nicotiana tabacum plants were studied. The rise in SOD and POX activities in the transformed lines TRa and TRb and the resulting regenerated plants and a decreased level of H₂O₂ in them as compared with the untransformed lines indicates that rol gene expression decreases H₂O₂ level probably by increasing production of antioxidant enzymes. A decreased H₂O₂ content in TRc line, in spite of similarity of antioxidant enzyme activity as compared to normal roots, indicates that rol genes activate other mechanisms except SOD and POX enzymes for reducing H₂O₂.