• 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.

• 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 Bio-Environment Control
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• v.17 no.4
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• pp.273-282
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• 2008

The relationship between water deficit stress and monodehydroascorbate reductase (MDHAR) activity was determined in lettuce (Lactuca sativa L.) leaves under water stress condition imposed by with-holding water for 72 hrs. Relative water content determined in water deficit stressed lettuce leaves gradually reduced from 91.29% to 74.58%, and water content of medium drastically decreased 4.73% after quitting of irrigation. Hydrogen peroxide content in leaves subjected to water deficit stress rapidly increased, but soluble protein content rapidly decreased when those were compared to control plant. The relationship between relative water content and hydrogen peroxide content in stressed leaves positively correlated with $R^2$=0.8851, but soluble protein content reversely correlated with $R^2$=0.9826. Total chlorophyll content in stressed plant leaves was higher than that of control plant, and increased rapidly in early stage of treatment of both stressed and control plants. Carotenoid content was higher than that of control plant, and the ratio of carotenoid to total chlorophyll in stressed plant was higher as compared to control plant. As water deficit stress continued progressively, total ascorbate content in stressed plant leaves was a little higher than that of control plant. But dehydroascorbate (DHA) content within 6 hr of water deficit stress was higher than that of control plant, and then, content of control plant in 12 hr of stress treatment higher than that of stressed leaves. The activity of monodehydroascorbate reductase of cytosolic and chloroplastic tractions increased dramatically, and mRNA of MDHAR was highly detected by probing $^{32}P$-labeled single stranded MDHAR RNA of lettuce plant leaves subjected to water deficit stress. Relationship between MDHAR activity and relative water content and hydrogen peroxide highly correlated with $R^2$=0.9937 and 0.8645, respectively.

• Journal of Plant Biology
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• v.38 no.1
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• pp.47-54
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• 1995

The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.3
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• pp.159-166
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• 2013

We have previously investigated the proteome changes of rice leaves under heat stress (Lee et al. in Proteomics 2007a, 7:3369-3383), wherein a group of antioxidant proteins and heat shock proteins (HSPs) were found to be regulated differently. The present study focuses on the biochemical changes and gene expression profiles of heat shock protein and antioxidant genes in rice leaves in response to heat stress ($42^{\circ}C$) during a wide range of exposure times. The results show that hydrogen peroxide and proline contents increased significantly, suggesting an oxidative burst and osmotic imbalance under heat stress. The mRNA levels of chaperone 60, HSP70, HSP100, chloroplastic HSP26, and mitochondrial small HSP responded rapidly and showed maximum expression after 0.5 or 2 h under heat stress. Transcript levels of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and Cu-Zn superoxide dismutase (Cu-Zn SOD) showed a rapid and marked accumulation upon heat stress. While prolonged exposure to heat stress resulted in increased transcript levels of monodehydroascorbate reductase, peroxidase, glyoxalase 1, glutathione reductase, thioredoxin peroxidase, 2-Cysteine peroxiredoxin, and nucleoside diphosphate kinase 1, while the transcription of catalase was suppressed. Consistent with their changes in gene expression, the enzyme activities of APX and DHAR also increased significantly following exposure to heat stress. These results suggest that oxidative stress is usually caused by heat stress, and plants apply complex HSP- and antioxidant-mediated defense mechanisms to cope with heat stress.

• Korean Journal of Environmental Agriculture
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• v.19 no.4
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• pp.309-313
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• 2000

To investigate the effects of different UV-B levels on growth and biochemical defense response in plants, cucumber plants were subjected to three levels of biologically effective ultraviolet-B $(UV-B_{BE})$ radiation [daily dose: 0.03 (No), 6.40 (Low) and $11.30\;(High)\;kJ{\cdot}m^{-2}$, $UV-B_{BE}$] in the growth chambers for 3 weeks during the early growth period. Enhanced UV-B radiation drastically decreased both dry weight and leaf area of cucumber. With increasing UV-B intensity, chlorophyll content was decreased, however the level of malondialdehyde was highly increased linearly. Total contents of ascorbic acid and glutathione were tended to increase by UV-B, while the ratios of dehydroascorbate/ascorbate and oxidized glutathione/reduced glutathione were significantly increased with increasing UV-B intensity in cucumber. All the enzyme activities investigated (superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, guaiacol peroxidase etc.) in cucumber were increased by the UV-B enhancement. These results suggested that enhanced UV-B irradiation caused photooxidative stress in cucumber plant and resulted in significant reduction in plant growth. Biochemical protection responses might be activated to prevent the leaves from damaging effects of oxidative stress generated by UV-B irradiation.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04a
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• pp.49-58
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• 2002

Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21s1 century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.1
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• pp.40-46
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• 2001

Ozone ($O_3$)-induced changes in chlorophyll content and specific activities of antioxidant enzymes, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) were investigated in two rice cultivars (Oryza sativa L.) grown under variable nutrient treatments. For this study, two rice cultivars of Ilpumbyeo (IL) and Keumobyeo#l (KM), which were known as resistant and susceptible to $O_3$, respectively, were exposed to $O_3$at 0.15ppm for 30 days and investigated with 10 days interval. The available nutrient regimes were varied by doubling the supply of nitrogen (N), phosphorus (P) and potassium (K) Within a basic fertilizer status (N, P, K; 15, 12, 12kg/l0a$^{-1}$ ). In both cultivars and at all nutrient status, chlorophyll content in $O_3$-treated plants decreased with prolonged treatment period, although higher N, P and K supply with $O_3$ treatment alleviated the decrease in chlorophyll content. The activities of almost all enzymes investigated for this study were decreased during initial stages of $O_3$- exposure except GPX which maintained higher activity throughout the exposure period than the non-treated plant. However, the antioxidant enzymes in $O_3$-treated plants showed almost the same or higher activities on 30 days after $O_3$ - exposure. The most significant changes in activities were observed in GR of the $O_3$-treated leaves. With the prolonged treatment period, the activity of GR at 30 days was increased by 3-8 times compared to those in 10 days. Most of the investigated enzymes showed very similar tendency to $O_3$ treatment in all fertilizer status. There was no observed evidence for enhanced detoxification of $O_3$-derived activated oxygen species in plants grown under higher fertilizer status compared with that in plants grown under basic fertilizer status. The increase in the activities of SOD, APX and GR in rice leaves by relatively long-term treatment with $O_3$ at low concentration is considered to indicate that the plant became adapted to the $O_3$ stress and the protection system increased its capacity to scavenge toxic oxygen species. Our results in two rice cultivars indicated that there was little difference in the activities of antioxidant enzymes between IL and KM, which were known as resistant and susceptible cultivar to $O_3$

• 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.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.49-58
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• 2002

Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21st century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.