• Title/Summary/Keyword: Ascorbate-glutathione cycle

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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.

Photochemical Response in 0-Year-Old and 1-Year-Old Needles of Picea glehnii during Cold Acclimation and Low Temperature

  • Bae, Jeong-Jin;Hara, Toshihiko;Choo, Yeon-Sik
    • Journal of Ecology and Environment
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    • v.31 no.4
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    • pp.317-325
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    • 2008
  • P. glehnii, an evergreen conifer found in northern areas, is known as a cold-resistant species. In this experiment, we measured the water content, PSⅡ efficiency, chlorophyll fluorescence, pigments of the xanthophyll-cycle and activity of enzymes of the ascorbate-glutathione cycle during cold acclimation and at subsequent low-temperature conditions to examine the importance of acclimation to cold tolerance. P. glehnii showed a decrease in PSⅡ efficiency (especially in Fv) during cold acclimation and at subsequent low temperatures. However, cold-acclimated needles showed higher PSⅡ efficiency at low temperatures than nonacclimated needles. In addition, 0-YON (first-year needles) showed an increase in $\beta$-carotene and lutein, while 1-YON (one-year-old needles) immediately developed an antioxidant mechanism in the ascorbate-gluthathione cycle as soon as they were exposed to low temperature and both 0-YON and 1-YON showed increased zeaxanthin and de-epoxidation ratios at continuous low temperature. Based on our results, we suggest that P. glehnii maintain PSⅡ efficiency at low temperature by effectively protecting the photosynthetic apparatus from photo-damage by rapid induction of an antioxidant mechanism in 1-YON and dissipation of excess energy by $\beta$-carotene and lutein in 0-YON.

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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Nitric oxide modulates antioxidant defense and the methylglyoxal detoxification system and reduces salinity-induced damage of wheat seedlings

  • Hasanuzzaman, Mirza;Hossain, Mohammad Anwar;Fujita, Masayuki
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.353-365
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    • 2011
  • The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

Effect of Ascorbate on the Arsenic Uptake, ROS-scavenging Capacity, and Antioxidant Homeostasis in Rice

  • Jung, Ha-il;Kong, Myung-Suk;Chae, Mi-Jin;Lee, Eun-Jin;Jung, Goo-Bok;Kim, Yoo-Hak
    • Korean Journal of Soil Science and Fertilizer
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    • v.51 no.2
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    • pp.90-100
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    • 2018
  • Environmental pollution with arsenic (As) in croplands causes agricultural and health problems worldwide. Rice is an important crop in South Korea, and many studies have evaluated the relationship between As and glutathione (GSH) to alleviate As uptake from the soil into plants. However, information about the relationship between As and ascorbate (AsA) in rice seedlings is still limited with regard to As phytotoxicity. We therefore investigated changes in reactive oxygen species (ROS) and antioxidant levels in rice (Oryza sativa L. cv 'Dasan') seedlings with toxic As and/or AsA application. The exposure of rice seedlings to $15{\mu}M$ As inhibited plant growth and resulted in increased contents of superoxide, hydrogen peroxide, and malondialdehyde, and induced As uptake by the roots and leaves. Application of AsA to As-exposed seedlings ameliorated As-induced oxidative stress by enhancing the capacity of AsA-GSH cycle in applied plants and increasing As transfer from the roots to leaves. These results suggest that AsA application alleviated As-induced oxidative damage by maintaining sufficient levels of AsA and GSH.

Scavenging Reactive Oxygen Species by Rice Dehydroascorbate Reductase Alleviates Oxidative Stresses in Escherichia coli

  • Shin, Sun-Young;Kim, Il-Sup;Kim, Yul-Ho;Park, Hyang-Mi;Lee, Jang-Yong;Kang, Hong-Gyu;Yoon, Ho-Sung
    • Molecules and Cells
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    • v.26 no.6
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    • pp.616-620
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    • 2008
  • Maintaining redox balance is one of the crucial requirements for a cell to endure stress from the outside. Dehydroascorbate reductase (DHAR; EC 1.8.5.1) plays an important role in the ascorbate-glutathione cycle; one of the major ROS scavenging systems in most known biological systems. A cDNA clone of the DHAR gene from Oryza sativa (OsDHAR) was isolated and overexpressed in Escherichia coli BL21 (DE3) strain from the pET-28a(+) expression vector. The OsDHAR transformed E. coli cells showed significantly higher DHAR activity and a lower level of ROS than the E. coli cells transformed by an empty pET-28a(+) vector. Also, the DHAR-overexpressing E. coli strain was more tolerant to oxidant- and heavy metal-mediated stress conditions than the control E. coli strain. The results suggest that the overexpressed rice DHAR gene effectively functions in a prokaryotic system and provide protection to various oxidative stresses.

Effect of sulfur on the cadmium transfer and ROS-scavenging capacity of rice (Oryza sativa L.) seedlings

  • Jung, Ha-il;Chae, Mi-Jin;Kong, Myung-Suk;Kang, Seong-Soo;Kim, Yoo-Hak
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.187-187
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    • 2017
  • Cadmium (Cd) pollution is rapidly increasing in worldwide due to industrialization and urbanization. In addition to its negative effects on the environment, Cd pollution adversely affects human health. Rice (Oryza sativa L.) is an important agricultural crop worldwide, including South Korea, and studies have examined its ability to alleviate Cd uptake from the soil into plants. However, information about the relationship between sulfur (S) and antioxidants in rice seedlings is still limited with regard to Cd phytotoxicity. We therefore investigated the changes in reactive oxygen species (ROS) and antioxidants in rice (Oryza sativa L. 'Dongjin') seedlings exposed to toxic Cd, S treatment, or both. The exposure of rice seedlings to $30{\mu}M$ Cd inhibited plant growth; increased the contents of superoxide, hydrogen peroxide, and malondialdehyde (MDA); and induced Cd uptake by the roots, stems, and leaves. Application of S to Cd-stressed seedlings decreased Cd-induced oxidative stress by increasing the capacity of the glutathione (GSH)-ascorbate (AsA) cycle, promoted S assimilation by increasing cysteine, GSH, and AsA contents in treated plants, and decreased Cd transfer from the roots to the stems and leaves. In conclusion, S application of plants under Cd stress promoted Cys and GSH biosynthesis and GSH-AsA cycle activity, thereby lowering the rate of Cd transfer to plant shoots and promoting the scavenging of the ROS that resulted from Cd toxicity, thus alleviating the overall Cd toxicity. Therefore, these results provide insights into the role of S in regulating the tolerance, uptake, and translocation of Cd in rice seedlings. The results of this study indicate that S application should have potential as a tool for mitigating Cd-stress in cereal crops, especially rice.

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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.