Transgenic tobacco plants overexpressing cotton glutathione S-transferase (GST) show enhanced resistance to methyl viologen (original) (raw)
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Planta, 2005
Transgenic cotton (Gossypium hirsutum L.) lines expressing the tobacco glutathione S-transferase (GST) Nt107 were evaluated for tolerance to chilling, salinity, and herbicides, antioxidant enzyme activity, antioxidant compound levels, and lipid peroxidation. Although transgenic seedlings exhibited ten-fold and five-fold higher GST activity under normal and salt-stress conditions, respectively, germinating seedlings did not show improved tolerance to salinity, chilling conditions, or herbicides. Glutathione peroxidase (GPX) activity in transgenic seedlings was 30% to 60% higher under normal conditions, but was not different than GPX activity in wild-type seedlings under salt-stress conditions. Glutathione reductase, superoxide dismutase, ascorbate peroxidase, and monodehydroascorbate reductase activities were not increased in transgenic seedlings under salt-stress conditions, while dehydroascorbate reductase activity was decreased in transgenic seedlings under salt-stress conditions. Transgenic seedlings had 50% more oxidized glutathione when exposed to salt stress. Ascorbate levels were not increased in transgenic seedlings under salt-stress conditions. Malondialdehyde content in transgenic seedlings was nearly double that of wild-type seedlings under normal conditions and did not increase under salt-stress conditions. These results show that expression of Nt107 in cotton does not provide adequate protection against oxidative stress and suggests that the endogenous antioxidant system in cotton may be disrupted by the expression of the tobacco GST.
Plant and Cell Physiology, 2000
Overexpression of a tobacco glutathione S-transferase with glutathione peroxidase activity (GST/GPX) in transgenic tobacco (Nicotiana tabacum L.) enhanced seedling growth under a variety of stressful conditions. In addition to increased GST and GPX activity, transgenic GST/GPXexpressing (GST+) seedlings had elevated levels of monodehydroascorbate reductase activity. GST+ seedlings also contained higher levels of glutathione and ascorbate than wildtype seedlings and the glutathione pools were more oxidized. Thermal or salt-stress treatments that inhibited the growth of wild-type seedlings also caused increased levels of lipid peroxidation. These treatments had less effect on the growth of GST+ seedling growth and did not lead to increased lipid peroxidation. Stress-induced damage resulted in reduced metabolic activity in wild-type seedlings while GST+ seedlings maintained metabolic activity levels comparable to seedlings grown under control conditions. These results indicate that overexpression of GST/GPX in transgenic tobacco seedlings provides increased glutathione-dependent peroxide scavenging and alterations in glutathione and ascorbate metabolism that lead to reduced oxidative damage. We conclude that this protective effect is primarily responsible for the ability of GST+ seedlings to maintain growth under stressful conditions.
The Plant Journal, 1999
Plant expression cassettes containing the Escherichia coli cysE gene alleles (encoding SAT) were constructed. After the Agrobacterium-mediated transformation of tobacco, we identi®ed stable transformed plants containing several-fold higher SAT activity in comparison to the control plant. Determination of nonprotein thiol contents indicated two-to threefold higher cysteine and glutathione levels in some of these transgenic plants. The maximal elevation of the cysteine level was about fourfold while that of GSH was about twofold higher than in the controls. The most striking physiological consequence of the modi®cation of sulfur metabolite levels in the transgenic plants, however, was their several-fold increased resistance to oxidative stress generated by exogenous hydrogen peroxide.
Expression of a Chimeric Tobacco Peroxidase Gene in Transgenic Tomato Plants
Journal of the American Society for Horticultural Science. American Society for Horticultural Science
Tomato plants (Lycopersicon esculentum cv. OH 7814) were transformed via Agrobacterium tumefaciens with a chimeric tobacco anionic peroxidase (EC 1.11.1.7) gene joined to the cauliflower mosaic virus (CaMV) 35S promoter. Transgenic plants obtained by selection on kanamycin were found to have more than five times the total leaf peroxidase activity of control plants. Transformed tomato plants chronically wilted upon reaching sexual maturity. Two independently selected transformants were self-fertilized, and progeny were obtained that were homozygous for the foreign gene. Isoelectric focusing gels stained for peroxidase activity revealed a new tomato leaf peroxidase isoenzyme with a pI of 3.75, which is similar to that seen in Nicotiana sylvestris L. Mature tomato fruit were found to have up to 1600-fold higher peroxidase activity in transformants expressing the tobacco anionic peroxidase (TobAnPOD) than control plants. Tissue blots showed the tobacco enzyme evenly distributed throughout the tomato fruit tissue. Progeny plants possessing the tobacco peroxidase gene (now homozygous) showed stunting, and fruit size was reduced by >80%. However, fruit set was normal and the rate of ripening was not altered from control plants. Fruit from transformed plants were found to have normal pigmentation, but the soluble solids concentration was 400% higher than in control tomato fruit. This result was predicted from the peroxidase-induced water stress. Possible roles for the tobacco anionic peroxidase in growth, development, and stress resistance are discussed.
Oxidative stress as DNA damage in different transgenic tobacco plants
One of the important mechanisms by which plant cells are damaged during adverse environmental conditions is the excessive production of reactive oxygen species (ROS). Major enzymatic ROS scavenging mechanisms of plants include superoxide dismutase (SOD) and catalase (CAT). The aim of this work was to assess leaf cell sensitivity to oxidative stress by measuring DNA damage (Comet assay) in Nicotiana tabacum wild type plants and transgenic lines with deregulated levels of CAT and MnSOD; DNA migration, as DNA damage measure, and nuclei with completely disintegrated head region (hedgehogs), as toxicity parameter, were recorded. A general time dependent increase of DNA migration values was observed within 3 h after hydrogen peroxide treatment, followed by an increase of hedgehog percentage. We showed that hydrogen peroxide treatments trigger different degrees of both DNA migration and hedgehog induction in the transgenic lines. There was only weak difference between the transgenic lines with increased MnSOD levels and wild type plants; but within CAT and MnSOD deficient plants, both levels of DNA migration and hedgehogs were significantly altered.
Plant Cell Reports, 2007
The effect of simultaneous expression of genes encoding three antioxidant enzymes, copper zinc superoxide dismutase (CuZnSOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), and dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1), in the chloroplasts of tobacco plants was investigated under oxidative stress conditions. In previous studies, transgenic tobacco plants expressing both CuZnSOD and APX in chloroplast (CA plants), or DHAR in chloroplast showed enhanced tolerance to oxidative stresses, such as paraquat and salt. In this study, in order to develop transgenic plants that were more resistant to oxidative stress, we introduced the gene encoding DHAR into CA transgenic plants. Mature leaves of transgenic plants expressing all three antioxidant genes (CAD plants) had approximately 1.6-2.1 times higher DHAR activity, and higher ratios of reduced ascorbate (AsA) to DHA, and oxidized glutathione (GSSG) to reduced glutathione (GSH) compared to CA plants. CAD plants were more resistant to paraquat-induced stress, exhibiting only 18.1% reduction in membrane damage relative to CA plants. In addition, seedlings of CAD plants had enhanced tolerance to NaCI (100 mM) compared to CA plants.