• 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.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.381-387
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• 1987

Distribution of superoxide dismutase (SOD) which catalyzes the dismutation of superoxide radicals to hydrogen peroxide and oxygen has been examined in various genera of bacteria. SOD was produced by various bacteria independent of genus and species with variation in superoxide dismutase activity of each bacteria. Bacillus circulans which produced relatively large amount of SOD was selected and used to investigate the optimum culture conditions and further studies. The compositions of optimum culture medium for the enzyme production were 1% glucose, 2% polypeptone, 0.l% NaCl, and 0.2mM of methyl viologen and initial pH was 6.0. The highest enzyme production was observed after 20 hours of cultivation at 3$0^{\circ}C$ on a reciprocal shaker. The enzyme activity was maintained stably for a relatively long period by the addition of 5% ethanol in pH 5.0, 0.01M acetate buffer.

• Journal of Applied Biological Chemistry
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• v.49 no.1
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• pp.1-7
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• 2006

Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.3
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• pp.223-230
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• 2017

Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) is an upstream signaling molecule that has been shown to induce stress tolerance in plants. In this study, the AtNDPK2 gene, under the control of a stress-inducible SWPA2 promoter, was introduced into the genome of tall fescue (Festuca arundinacea Schreb.) plants. The induction of the transgene expression mediated by methyl viologen (MV) and NaCl treatments were confirmed by RT-PCR and northern blot analysis, respectively. Under salt stress treatment, the transgenic tall fescue plants (SN) exhibited lower level of $H_2O_2$ and lipid peroxidation accumulations than the non-transgenic (NT) plants. The transgenic tall fescue plants also showed higher level of NDPK enzyme activity compared to NT plants. The SN plants were survived at 300 mM NaCl treatment, whereas the NT plants were severely affected. These results indicate that stress-inducible overexpression of AtNDPK2 might efficiently confer the salt stress tolerance in tall fescue plants.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.405-410
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• 2003

Environmental stress is the major limiting factor in plant productivity. In order to evaluate the stress tolerance of potato plants, leaf discs of two potato cultivars, Atlantic and Superior, were subjected to various stress conditions of high temperature, methyl viologen, H2O2, or $H_2O$$_2$. When potato leaf discs were exposed to high temperature at 37$^{\circ}C$ for 84 hr, Atlantic plants, a cultivar with high sensitivity to heat stress, showed about 20％ higher membrane damage than Superior plants. When exposed to 2$\mu$M methyl violgen (MV), a superoxide generating non-selective herbicide, for 36 hr, Atlantic plants also showed about 38％ higher membrane damage than Superior plants, and were more susceptible up to 10$\mu$M MV concentration tested. On treatment with 0.75M NaCl, Atlantic plants also had about 45％ less chlorophyll contents in leaf discs than Superior plants. There was, however, no difference in chlorophyll content of two cultivars at higher NaCl concentrations. The effect of $H_2O$$_2$ on the two cultivars was mixed. At low $H_2O$$_2$ concentration (25 mM) , Superior plants were more susceptible to $H_2O$$_2$stress after 36 hr. However, at high $H_2O$$_2$ concentration (100 mM), Atlantic plants exhibited higher susceptibility after 36 hr. The results indicate that in vitro leaf discs reflecting the whole plants in this study will be useful for selection and characterization of elite transgenic potato plants with enhanced tolerance to environmental stress.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.717-724
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• 2003

A newly isolated Citrobacter sp. Y19 catalyzes the CO-dependent $H_2$ production (biological water-gas shift reaction) by the actions of CO dehydrogenase (CODH) and hydrogenase. Y 19 requires $O_2$ for fast growth, but its $H_2$ production activity is significantly inhibited by $O_2$. In the present study, the effect of $O_2$ on the activities of CODH ard hydrogenase was investigated quantitatively in both whole cells and broken cells, based on CO-dependent or methyl viologen (MV)-dependent $H_2$ production in addition to CO-dependent MV reduction. In crude cell extracts, CODH activity was mostly found in the soluble fraction. Inactivation of CODH and hydrogenase activities by $O_2$ followed the first-order decay kinetics, and the dependence of the rate constants on $O_2$ partial pressure could be expressed by the Michaelis-Menten equation. In whole cells, the maximum deactivation rate constants ($k_{d,max}$ of hydrogenase and CODH were quite similar: $0.07{\pm}0.03 min^{-1}\;and\;0.10{\pm}0.04 min^{-1}$, respectively. However, the first-order rate constant ($k_{d,max}/K_s$) of CODH ($0.25\;min^{-1}\;atm^{-1}$) at low $O_2$ partial pressures was about 3-fold higher than that of the hydrogenase, since the half-saturation constant ($K_s$) of CODH was about half of that of hydrogenase. In broken cells, both enzymes became significantly more sensitive to $O_2$ compared to the unbroken cells, while $k_{d,max}/K_s$ increased 37-fold for hydrogenase and 6.7-fold for CODH. When whole cells were incubated under anaerobic conditions after being exposed to air for 1 h, hydrogenase activity was recovered more than 90% in 2 h suggesting that the deactivation of hydrogenase by $O_2$ was reversible. On the contrary, CODH activity was not recovered once deactivated by $O_2$ and the only way to recover the activity was to synthesize new CODH. This study indicates that $O_2$ sensitivity of $H_2$ production activity of Citrobacter sp. Y19 is an important drawback as in other $H_2-producing$ bactria.

• 한국정보디스플레이학회:학술대회논문집
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• 2008.10a
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• pp.993-994
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• 2008

Electrochromic (EC) devices are capable of reversibly changing their optical properties upon charge injection and extraction induced by the external voltage. The characteristics of the EC device, such as low power consumption, high coloration efficiency, and memory effects under open circuit status, make them suitable for use in a variety of applications including smart windows and electronic papers. Coloration due to reduction or oxidation of redox chromophores can be used for EC devices (e-paper), but the switching time is slow (second level). Recently, with increasing demand for the low cost, lightweight flat panel display with paper-like readability (electronic paper), an EC display technology based on dye-modified $TiO_2$ nanoparticle electrode was developed. A well known organic dye molecule, viologen, was adsorbed on the surface of a mesoporous $TiO_2$ nanoparticle film to form the EC electrode. On the other hand, ZnO is a wide bandgap II-VI semiconductor which has been applied in many fields such as UV lasers, field effect transistors and transparent conductors. The bandgap of the bulk ZnO is about 3.37 eV, which is close to that of the $TiO_2$ (3.4 eV). As a traditional transparent conductor, ZnO has excellent electron transport properties, even in ZnO nanoparticle films. In the past few years, one-dimension (1D) nanostructures of ZnO have attracted extensive research interest. In particular, 1D ZnO nanowires renders much better electron transportation capability by providing a direct conduction path for electron transport and greatly reducing the number of grain boundaries. These unique advantages make ZnO nanowires a promising matrix electrode for EC dye molecule loading. ZnO nanowires grow vertically from the substrate and form a dense array (Fig. 1). The ZnO nanowires show regular hexagonal cross section and the average diameter of the ZnO nanowires is about 100 nm. The cross-section image of the ZnO nanowires array (Fig. 1) indicates that the length of the ZnO nanowires is about $6\;{\mu}m$. From one on/off cycle of the ZnO EC cell (Fig. 2). We can see that, the switching time of a ZnO nanowire electrode EC cell with an active area of $1\;{\times}\;1\;cm^2$ is 170 ms and 142 ms for coloration and bleaching, respectively. The coloration and bleaching time is faster compared to the $TiO_2$ mesoporous EC devices with both coloration and bleaching time of about 250 ms for a device with an active area of $2.5\;cm^2$. With further optimization, it is possible that the response time can reach ten(s) of millisecond, i.e. capable of displaying video. Fig. 3 shows a prototype with two different transmittance states. It can be seen that good contrast was obtained. The retention was at least a few hours for these prototypes. Being an oxide, ZnO is oxidation resistant, i.e. it is more durable for field emission cathode. ZnO nanotetropods were also applied to realize the first prototype triode field emission device, making use of scattered surface-conduction electrons for field emission (Fig. 4). The device has a high efficiency (field emitted electron to total electron ratio) of about 60%. With this high efficiency, we were able to fabricate some prototype displays (Fig. 5 showing some alphanumerical symbols). ZnO tetrapods have four legs, which guarantees that there is one leg always pointing upward, even using screen printing method to fabricate the cathode.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.151-159
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• 2005

Injury caused by reactive oxygen species (ROS), known as oxidative stress, is one of the major damaging factors in plants exposed to environmental stress. Chloroplasts are specially sensitive to damage by ROS because electrons that escape from the photosynthetic electron transfer system are able to react with relatively high concentration of $O_2$ in chloroplasts. To cope with oxidative stress, plants have evolved an efficient ROS-scavenging enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX), and low molecular weight antioxidants including ascorbate, glutathione and phenolic compounds. To maintain the productivity of plants under the stress condition, it is possible to fortify the antioxidative mechanisms in the chloroplasts by manipulating the antioxidation genes. A powerful gene expression system with an appropriate promoter is key requisite for excellent stress-tolerant plants. We developed a strong oxidative stress-inducible peroxidase (SWPA2) promoter from cultured cells of sweetpotato (Ipomoea batatas) as an industrial platform technology to develop transgenic plants with enhanced tolerance to environmental stress. Recently, in order to develop transgenic sweetpotato (tv. Yulmi) and potato (Solanum tuberosum L. cv. Atlantic and Superior) plants with enhanced tolerance to multiple stress, the genes of both CuZnSOD and APX were expressed in chloroplasts under the control of an SWPA2 promoter (referred to SSA plants). As expected, SSA sweetpotato and potato plants showed enhanced tolerance to methyl viologen-mediated oxidative stress. In addition, SSA plants showed enhanced tolerance to multiple stresses such as temperature stress, drought and sulphur dioxide. Our results strongly suggested that the rational manipulation of antioxidative mechanism in chloroplasts will be applicable to the development of all plant species with enhanced tolerance to multiple environmental stresses to contribute in solving the global food and environmental problems in the 21st century.

• Journal of The Korean Society of Grassland and Forage Science
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• v.27 no.2
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• pp.109-116
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• 2007

Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.