• BMB Reports
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• v.34 no.4
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• pp.316-321
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• 2001

Dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1) catalyzes the reduction of DHA to reduced ascorbate (AsA) using glutathione (GSH) as the electron donor in order to maintain an appropriate level of ascorbate in plant cells. To analyze the physiological role of DHAR in environmental stress adaptation, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants that express a human DHAR gene isolated from the human fetal liver cDNA library in the chloroplasts. We also investigated the DHAR activity, levels of ascorbate, and GSH. Two transgenic plants were successfully developed by Agrobacterium-mediated transformation and were confirmed by PCR and Southern blot analysis. DHAR activity and AsA content in mature leaves of transgenic plants were approximately 1.41 and 1.95 times higher than in the non-transgenic (NT) plants, respectively In addition, the content of oxidized glutathione (GSSG) in transgenic plants was approximately 2.95 times higher than in the NT plants. The ratios of AsA to DHA and GSSG to GSH were changed by overexpression of DHAR, as expected, even though the total content of ascorbate and glutathione was not significantly changed. When tobacco leaf discs were subjected to methyl viologen at $5\;{\mu}M$, $T_0$ transgenic plants showed about a 50% reduction in membrane damage compared to the NT plants.

• The Plant Pathology Journal
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• v.17 no.2
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• pp.88-93
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• 2001

Oxidative stress is one of the major limiting factor in plant productivity. Reactive oxygens species (ROS) generated during metabolic processes damage cellular functions and consequently lead to disease, senescence and cell death. Plants have evolved an efficient defense system by which the ROS is scavenged by antioxidant enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX). Attempts to reduce oxidative damages under the stress conditions have included the manipulation of 갠 scavenging enzymes by gene transfer technology. Increased SOD activities of transgenic plants lead to increased resistance against oxidative stresses derived from methyl viologen (MV), and from photooxidative damage caused by high light and low temperature. Transgenic tobacco plants overexpressing APX showed reduced damage following either MV treatment of photooxidative treatment. Overexpression of glutathion reductase (GR) leads to increase in pool of ascorbate and GSH, known as small antioxidant molecules. These results indicate through overexpression of enzymes involved in ROS-scavenging could maintain or improve the plant productivities under environment stress condition. In this study, the rational approaches to develop stress-tolerant plants by gene manipulation of antioxidant enzymes will be introduced to provide solutions for the global food and environmental problems in the $21^\textrm{st}$ century.

• Korean Journal of Soil Science and Fertilizer
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• v.32 no.2
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• pp.164-170
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• 1999

This experiment was carried out to elucidate change of antioxidant enzymes activities subjected to water stresses in soybean plant. In this study, we measured the activities of ascorbate peroxidase(APDX), monodehydroascorbate reductase(MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase(GR) subjected to drought or flooding stresses for 4days and following recovery for 3days. Leaves of two soybean lines subjected to drought or flooding showed premature senescence as evidenced by the decrease in water content and total soluble protein content but those of soybean leaves was increased when stresses were recovered for 3days. The activities of APDX and GR subjected to drought or flooding were the decrease but those of enzymes were recovered when water stress was recovered. The activities of MDHAR with drought or flooding were on the decrease, whereas those of DHAR were increased, respectively. Antioxidant contents decreased continually subjected to drought or flooding but it recovered after 3 days subjected to water stresses.

• Journal of Korean Society of Forest Science
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• v.96 no.1
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• pp.14-20
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• 2007

In order to examine the differences in antioxidant enzyme activities which represent defence mechanism to stressful environments, and soil properties between healthy and declining (or unhealthy) trees, we selected three sites, Witseorum, Youngsil and Sungpanak (Jindallebat). Antioxidant enzymes including Ascorbate peroxidase (APX) and Glutathione Reductase (GR), forest soil properties including soil texture, soil pH, organic matter, total nitrogen, available phosphate, cation exchange capacity, exchangeable cation content and nutrient contents in leaves of Abies koreana (Korean fir) trees were analyzed. There were no significant differences between healthy and declining trees in GR activity. However, seasonal difference in antioxidant enzyme activity was observed. GR activity was lower in June and August than that of September. Soil chemical and physical properties of each site showed a tendency that organic content, total nitrogen content, available phosphorus, cation exchange capacity and cation content were lower at the site of declining trees than the site of healthy trees.

• 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.

• Journal of Plant Biotechnology
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• v.43 no.2
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• pp.223-230
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• 2016

Ascorbic acid (AsA) is a strong antioxidant/reducing agent that can be converted to dehydroascorbate (DHA) by oxidation in plants. DHA, a very short-lived chemical, is recycled to AsA by dehydroascorbate reductase (DHAR). Previously, DHAR cDNA was isolated from the hairy roots of the sesame plant, and DHAR-overexpressing transgenic potato plants were generated under the control of the CaMV35S promoter (CaMV35S::DHAR). An increase in transgene expression and ascorbate levels were observed in the transgenic plants. In the present study, proteomic analysis revealed that transgenic plants not only accumulated DHAR in their cells, but also induced several other antioxidant enzyme-related proteins during plant growth. These results suggest that DHAR is important for stress tolerance via induction of antioxidant proteins, and could improve stress tolerance in transgenic potato plants.

• 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.

• Journal of Life Science
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• v.18 no.12
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• pp.1705-1711
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• 2008

To investigate the relationship between cold stress and the activity of ascorbate peroxidase(APX), dehydroascorbate reductase (DHAR), mRNA expression level of two enzymes, hydrogen peroxide content was studied in lettuce leaves under stress condition imposed by cold stress at $4^{\circ}C$ for 24 hr in the dark and following recovery at $20^{\circ}C$ from cold stress. Hydrogen peroxide content increased gradually in lettuce leaves during cold stress, but decreased slightly following recovery from cold stress. Soluble protein content, however, decreased gradually during cold stress, and then rapidly returned to normal levels following recovery. Total chlorophyll content decreased gradually during cold stress, and then keep constant following recovery. The patterns of chlorophyll a and b content similar to that of total chlorophyll content, and carotenoid content didn't change. The ratio of chlorophyll a and total chlorophyll was increased during cold stress, but decreased with rapid during cold stress, and then the ratio returned to normal levels following recovery. During cold stress, the activity of APX and DHAR in the lettuce leaves increased dramatically, and also transcript levels of mRNA of APX and DHAR, as determined by probing 32P-labeled single stranded RNA of APX and DHAR, highly increased and returned to normal levels following recovery, respectively. Relationship between APX and DHAR activity and hydrogen peroxide highly related ($R^2$=0.8715 and 0.8643), whereas between hydrogen peroxide and total chlorophyll content and soluble content related reversely ($R^2$=0.5021 and 0.8915).

• Applied Biological Chemistry
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• v.42 no.3
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• pp.246-251
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• 1999

This experiment was carried out to elucidate and investigate the changes in the content of antioxidants and the activities of antioxidant enzymes in the leaves of soybean subjected to water stresses. The results obtained were as follows; Leaves of soybeans subjected to water stresses have showed the differences in the activities of the antioxidant enzymes. In eunhakong, the activity of APOX was increased within a few days, but that of GR was decreased, whereas the activities of APOX and GR were gradually decreased in eunhakong. The activity of MDHAR of the leaves of eunhakong subjected to drought stress was gradually increased within 4days, whereas that of flooding was increased within 2days. We are supposed that the activities of APOX and MDHAR are coupled to maintain ascorbate concentration. In eunhakong, the relative activity of DHAR subjected to flooding was higher than that of drought. These results imply that DHAR is the only enzyme participating in the regeneration of ascorbate when the activity of MDHAR was limited by the deficiency of NADPH. The contents of ascorbate and reduced glutathione subjected to drought stress decreased continually, whereas those subjected to flooding stress recovered after five days of treatment.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.235-235
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• 2017

Chromium (Cr) toxicity is hazardous to the seed germination, growth, and development of plants. ${\gamma}$-Aminobutyric acid (GABA) is a non-protein amino acid and is involved in stress tolerance in plants. To investigate the effects of GABA in alleviating Cr toxicity, we treated eight-d-old mustard (Brassica juncea L.) seedlings with Cr (0.15 mM and 0.3 mM $K_2CrO_4$, 5 days) alone and in combination with GABA ($125{\mu}M$) in a semi-hydroponic medium. The roots and shoots of the seedlings accumulated Cr in a dose-dependent manner, which led to an increase in oxidative damage [lipid peroxidation; hydrogen peroxide ($H_2O_2$) content; superoxide ($O{_2}^{{\cdot}-}$) generation; lipoxygenase (LOX) activity], MG content, and disrupted antioxidant defense and glyoxalase systems. Chromium stress also reduced growth, leaf relative water content (RWC), and chlorophyll (chl) content but increased phytochelatin (PC) and proline (Pro) content. Furthermore, supplementing the Cr-treated seedlings with GABA reduced Cr uptake and upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH) and the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II), and finally reduced oxidative damage. Adding GABA also increased leaf RWC and chl content, decreased Pro and PC content, and restored plant growth. These findings shed light on the effect of GABA in improving the physiological mechanisms of mustard seedlings in response to Cr stress.