• Title/Summary/Keyword: Ascorbate-glutathione cycle enzyme

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Effects of nitric oxide on ascorbate-glutathione cycle enzymes activities in chinese cabbage leaves under paraquat-induced oxidative stress (Paraquat 유도 산화스트레스하의 배추 잎에서 Ascorbate-Glutathione 회로 효소의 활성도에 대한 산화질소 (Nitric oxide)의 효과)

  • Na, Ho-Gyun;Jin, Chang-Duck
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.73-80
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    • 2014
  • Pretreatment of chinese cabbage leaves with $100{\mu}M$ sodium nitroprusside (SNP), a nitric oxide (NO) donor, effectively improved their tolerance to subsequent $2{\mu}M$ paraquat (PQ)-induced oxidative damage. The fresh weight, and chlorophyll and protein contents in primary leaves treated with PQ alone were noticeably reduced over 24 h light incubation. However, these leaf injury symptoms were significantly alleviated with $100{\mu}M$ SNP pretreatment for 3 h prior to PQ exposure. In additions, the increase of the contents of malondialdehyde (MDA) and $H_2O_2$ due to PQ exposure were significantly inhibited by SNP pretreatment. Together with the protective effects of SNP against PQ toxicity in leaves, the changes of ascorbate-glutathione cycle enzymes activities were examined. In the PQ alone treatment, the activities of APX, DHAR, and GR after 6 h incubation were rapidly reduced and showed 19%, 50% and 39% respectively, compared with those of the control. However, the decreases in these enzyme activities were significantly inhibited by SNP pretreatment. As a result, their activities were higher than those of PQ alone treatment by 5 times, 2 times, and 1.5 times, respectively, at 6 h incubation. Thereafter, these enzymes decrease their activities gradually showing high levels than those of PQ alone. Based on the above results, it can be assumed that the activation of ascorbate-glutathione cycle by SNP pretreatment in chinese cabbage leaves exposed to PQ can prevent $H_2O_2$ accumulation, thereby leading to protection against PQ-induced oxidative stress. Also, these results indicate that NO acts as an protectant against PQ stress in the leaves of chinese cabbage.

Changes in element accumulation, phenolic metabolism, and antioxidative enzyme activities in the red-skin roots of Panax ginseng

  • Zhou, Ying;Yang, Zhenming;Gao, Lingling;Liu, Wen;Liu, Rongkun;Zhao, Junting;You, Jiangfeng
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.307-315
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    • 2017
  • Background: Red-skin root disease has seriously decreased the quality and production of Panax ginseng (ginseng). Methods: To explore the disease's origin, comparative analysis was performed in different parts of the plant, particularly the epidermis, cortex, and/or fibrous roots of 5-yr-old healthy and diseased red-skin ginseng. The inorganic element composition, phenolic compound concentration, reactive oxidation system, antioxidant concentrations such as ascorbate and glutathione, activities of enzymes related to phenolic metabolism and oxidation, and antioxidative system particularly the ascorbate-glutathione cycle were examined using conventional methods. Results: Aluminum (Al), iron (Fe), magnesium, and phosphorus were increased, whereas manganese was unchanged and calcium was decreased in the epidermis and fibrous root of red-skin ginseng, which also contained higher levels of phenolic compounds, higher activities of the phenolic compound-synthesizing enzyme phenylalanine ammonia-lyase and the phenolic compound oxidation-related enzymes guaiacol peroxidase and polyphenoloxidase. As the substrate of guaiacol peroxidase, higher levels of $H_2O_2$ and correspondingly higher activities of superoxide dismutase and catalase were found in red-skin ginseng. Increased levels of ascorbate and glutathione; increased activities of $\text\tiny L$-galactose 1-dehydrogenase, ascorbate peroxidase, ascorbic acid oxidase, and glutathione reductase; and lower activities of dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione peroxidase were found in red-skin ginseng. Glutathione-S-transferase activity remained constant. Conclusion: Hence, higher element accumulation, particularly Al and Fe, activated multiple enzymes related to accumulation of phenolic compounds and their oxidation. This might contribute to red-skin symptoms in ginseng. It is proposed that antioxidant and antioxidative enzymes, especially those involved in ascorbate-glutathione cycles, are activated to protect against phenolic compound oxidation.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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The Responses of Antioxidative Enzymes and Salt Tolerance of Atriplex gmelini (Atriplex gmelini(가는갯능쟁이)의 내염성과 항산화 효소 반응)

  • 배정진;윤호성;추연식;송승달
    • The Korean Journal of Ecology
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    • v.26 no.5
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    • pp.273-280
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    • 2003
  • Saline conditions invoke oxidative stress attributed to the overproduction of reactive oxygen species (ROS). Changes in quantum efficiency and antioxidative enzyme activity upon salt treatment were examined in a salt-tolerant plant, Atriplex gmelini, to test the hypothesis that salt tolerance of A. gmelini is due to the increased activity of antioxidative enzymes. A. gmelini showed optimum growth at 100 mM NaCl producing 116% of the shoot dry weight over control plants in 0 mM NaCl treatment. Healthy growth persisted up to 300 mM NaCl treatment maintaining normal internal water content and dry weight. No photochemical stress or damages on antioxidative defense system was obvious in plants of 2 and 4 day salt treatment which was indicated by increased quantum efficiency (Fv/Fm value), decreased stress index (Fo/Fm value), and increased activity of antioxidative enzymes such as SOD, APX, GR. However, the plants treated with 400 mM NaCl showed decrease in growth and in antioxidative enzyme activity although the enzyme activity was still higher than that of the 0 mM NaCl treated plants (l31%, 114%, and 134% of the SOD, APX, and GR activity, respectively). Interestingly, another important antioridative enzyme that scavenges H₂O₂ in plant cells, CAT, showed rapid decrease in its activity as salt concentration increased; 38%, 22%, 15% of the 0 mM NaCl treated plants at 200, 300, 400 mM NaCl treatments, respectively. It appears that the enzymes in ascorbate-glutathione cycle such as APX and GR play the major roles in scavenging ROS produced by salt stress in A. gmelini. After 6 days of salt treatment, the damage in photochemical and antioxidative defense system was indicated by decreased Fv/Fm value and increased Fo/Fm value. A. gmelini appears to cope with short term salt treatment by enhanced activity of the antioxidative defense system, whereas long term stress invoke oxidative stress by increased ROS due to the damages in photochemical and antioxidative system.