• Journal of Korean Society of Forest Science
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• v.96 no.5
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• pp.551-560
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• 2007

We investigated physiological damages and biochemical alleviation of five species of genus Acer under ozone fumigation in order to assess their tolerant ability against ozone toxicity. At the end of 150 ppb $O_3$ fumigation, photosynthetic characteristics were measured, and chlorophyll contents, malondialdehyde (MDA) and antioxidative enzyme activities were analyzed in the leaves of five maple trees (Acer buergerianum, A. ginnala, A. mono, A. palmatum, and A. palmatum var. sanguineum). The reduction of chlorophyll (chl) a in ozone-exposed plants was 16.8% (A. buergerianum) to 26.7% (A. ginnala) of control plants. For the content of chi b, A. ginnala and A. palmatum var. sanguineum represented the high reduction of 26.3% and 23.6%, respectively. The highest reduction on the chi a:b ratio was observed in the leaves of A. palmatum. The reduction of net photosynthesis in five species varied from 2.4% to 37.6%. Among five species, A. ginnala showed remarkable reduction (37.6%) for net photosynthesis in comparison with control. Carboxylation efficiency differed significantly (P < 0.05) among species and between control and ozone treatment. The reduction of carboxylation efficiency was the highest in the leaves of A. ginnala (44.7%). A. palmatum var. sanguineum showed the highest increase (41.7%) for MDA content. The highest increase of superoxide dismutase (SOD) activity represented in A. palmatum (26.1%) and the increase of ascorbate peroxidase (APX) activity ranged from 16.5% (A. ginnala) to 49.1% (A. palmatum var. sanguineum). A. mono showed the highest increase (376.6%) of glutathione reductase (GR) activity under ozone fumigation and A. buergerianum also represented high increase (42.3%) of GR activity. Catalse (CAT) activity increased in the leaves of A. ginnala, A. palmatun and A. palmatum var. sanguineum under ozone exposure, whereas A. buergerianum and A. mono decreased in comparison with control plants. In conclusion, physiological markers such as chlorophyll content and photosynthesis that responded sensitively to $O_3$ in maple trees were considered as the very important indicators in order to evaluate the tolerance against $O_3$ stress, and parameters were closely related with each other. Among anti oxidative enzymes, SOD and APX might be contributed to alleviate to $O_3$ toxicity through the increase of activity in all maple trees. Therefore, these compounds can be used as a biochemical maker to assess the stress tolerance to $O_3$.

• Journal of Korean Society of Forest Science
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• v.97 no.5
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• pp.508-515
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• 2008

Sycamore (Platanus occidentalis L.) seedlings were grown under low light intensity and ozone treatments to investigate the role of the light environment in their response to chronic ozone stress. One-year-old seedlings of Platanus occidentalis L. were grown in pots for 3 weeks under low light (OL, $150{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) and high light (OH, $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) irradiance in combination with 150 ppb of ozone fumigation. After three weeks of ozone and light treatment, seedlings were placed in ozone free clean chamber for 3 weeks for recovery from ozone stress with same light conditions to compare recovery capacity. Ozone fumigation determined an impairment of the photosynthetic process. Reduction of leaf dry weight (14%) and shoo/root ratio (17%) were observed in OH treatment. OL treatment also showed severe reductions in leaf dry weight and shoot/root ratio by 48% and 36% comparing to control, respectively. At the recovery phase, OH-treated plants recovered their biomass, whereas OL-treated plant showed reduction in leaf dry weight (52%) and shoot/root ratio (49%). OH-treated plants reached similar relative growth rate (RGR) comparing to control, whereas OL-treated plants showed lower RGR in stem height. However, there were no significant differences in response to those treatments in stem diameter RGR at the recovery phase. Ozone treatment produced significant reduction of net photosynthesis in both high and low light treatments. Carboxylation efficiency and apparent quantum yield in OL-treated plants showed significant reductions rate to 10% and 45%, respectively. At the recovery stage, ozone exposed seedlings under high light had similar photosynthetic capacity comparing to control plants. Antioxidant enzymes activities such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR) were increased in ozone fumigated plants only under low light. The present work shows that the physiological changes occur in photosynthesis-related parameters and growth due to ozone and low light stress. Thus, low light seems to enhance the detrimental effects of ozone on growth, photosynthesis, and antioxidant enzyme responses.

• Korean Journal of Agricultural and Forest Meteorology
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• v.11 no.1
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• pp.3-12
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• 2009

Ozone tolerance of tree species was determined by standard index of physiological damages and biochemical defense responses under short-term ozone exposure. At the end of 150ppb $O_3$ fumigation, photosynthetic characteristics and antioxidative enzyme activities were analyzed in the leaves of five species(Koelreuteria paniculata, Firmiana simplex, Styrax japonica, Fraxinus rhynchophylla, Viburnum sargentii). Injury index was determined by the effect of ozone on photosynthetic parameters and malondialdehyde(MDA) content, and tolerance index was calculated using the rate of increase in superoxide dismutase(SOD), ascorbate-peroxidase(APX), glutathione reductase(GR) and catalase(CAT) activities. Apparent quantum yield(AQY), carboxylation efficiency(Ce) and photo-respiration rate(PR) decreased in the leaves of five species with increasing ozone exposure time. These parameters were considered as an appropriate indicator for stress evaluation. Antioxidative enzyme activities showed various results depending on the tree species, exposure time, and enzyme types. SOD activity of K. paniculata increased with ozone exposure time, and that of F. rhynchophylla increased only after 6 hours of ozone exposure. CAT activity of $O_3$-exposed F. simplex was lower than the control. Based on standard index, ozone tolerance ability of five species was determined as two tolerant species(F. rhynchophylla > K. paniculata) and three sensitive species(S. japonica > F. simplex > V. sargentii).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.133-138
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• 2006

To differentiate barley responses to drought and salt stress, barley seedlings at the second leaf stage were treated with 218 mM NaCl and 29.5% PEG6000 iso-osmotic to 218 mM NaCl for 6 days. Shoot fresh weight and leaf relative water content of barley seedlings were more reduced by drought compared to salt stress. Hydrogen peroxide content increased under both stress conditions, but its accumulation was more severe at 6 days after salt stress. The activity of ascorbate peroxidase, glutathione reductase (GR) and catalase (CAT) was enhanced until 4 days after salt stress. On the other hand, the activity of GR and CAT increased gradually until 6 days after drought. Among the amino acids measured in this study, the accumulation of glycine, arginine and GABA (${\gamma}-aminobutyric$ acid) was lower under salt stress than drought. However, considerably larger amount of proline was accumulated by salt stress. It is concluded that the antioxidant enzymes activity and amino acid content of barley seed-lings were differently regulated in response to the isoosmotic condition of salt and drought stress.

• BMB Reports
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• v.38 no.2
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• pp.218-224
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• 2005

Antioxidant enzymes are related to the resistance to various abiotic stresses including salinity. Barley is relatively tolerant to saline stress among crop plants, but little information is available on barley antioxidant enzymes under salinity stress. We investigated temporal and spatial responses of activities and isoform profiles of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), non-specific peroxidase (POX), and glutathione reductase (GR) to saline stress in barley seedlings treated with 200 mM NaCl for 0, 1, 2, 5 days, respectively. In the control plant, hydrogen peroxide content was about 2-fold higher in the root than in the shoot. Under saline stress, hydrogen peroxide content was decreased drastically by 70% at 2 d after NaCl treatment (DAT) in the root. In the leaf, however, the content was remained unchanged by 2 DAT and increased about 14 % at 5 DAT. In general, the activities of antioxidant enzymes were increased in the root and shoot under saline stress. But the increase was more significant and consistent in the root. The activities of SOD, CAT, APX, POX, and GR were increased significantly in the root within 1 DAT, and various elevated levels were maintained by 5 DAT. Among the antioxidant enzymes, CAT activity was increased the most drastically. The significant increase in the activities of SOD, CAT, APX, POX, and GR in the NaCl-stressed barley root was highly correlated with the increased expression of the constitutive isoforms as well as the induced ones. The hydrogen peroxide content in the root was most highly correlated with the CAT activity, indicating an increased role of CAT in hydrogen peroxide detoxification under salinity stress. In addition, the results suggest the significance of temporal and spatial regulation of each antioxidant isoform in determining the competence of the antioxidant capacity under saline stress.

• Journal of Life Science
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• v.13 no.5
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• pp.658-667
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• 2003

Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.

• 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.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.69-77
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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 (lpomoea 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 Weed Science
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• v.31 no.2
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• pp.134-145
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• 2011

The objectives of this study were to investigate fitness difference in growth and rice yield in herbicide-transgenic rice overexpressing Myxococcus xanthus and Arabidopsis thaliana protoporphyrinogen oxidase (Protox) genes and non-transgenic rice. We also aimed to determine whether these fitness differences are related to ALA synthesizing capacity, accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes at different growth stages of rice. Plant height of the transgenic rice overexpressing M. xanthus (MX) and A. thaliana (AP37) Protox genes at 43, 50, and 65 days after transplanting (DAT) was significantly lower than that of WT. Number of tiller of PX as well as MX and AP37 at 50 and 65 DAT was significantly lower than that of WT. At harvest time, culm length and yield of MX, PX and AP37 and rice straw weight of MX and AP37 were significantly low compared with WT. The reduction of yield in MX, PX, and AP37 was caused by spikelets per panicle and 1000 grain weight, ripened grain, spikelets per panicle, 1000 grain weight, and ripened grain, respectively. On the other hand, 135 the reduction of yield in MX, PX, and AP37 was also observed in another yearly variation experiment. The reduction of rice growth in MX, PX, and AP37 was observed in seedling stage as well as growth duration in field. There were no differences in tetrapyrrole intermediate Proto IX, Mg-Proto IX and Mg-Proto IX monomethyl ester, reactive oxygen species ($H_2O_2$ and ${O_2}^-$), MDA, antioxidative enzymes (SOD, CAT, POX, APX, and GR) and chlorophyll between transgenic lines and wild type, indicating that accumulated tetrapyrrole intermediate and other parameters were not related to growth reduction in transgenic rice. However, ALA synthesizing capacity in MX, PX, and AP37 at one day after exposure to light and 52 DAT was significantly lower than that of WT. Further study is required to elucidate the mechanisms underlying the growth and yield difference between transgenic and WT lines.

• Korean Journal of Agricultural and Forest Meteorology
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• v.12 no.3
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• pp.207-216
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• 2010

Individual differences of ozone ($O_3$) resistance for seed production, seed germination and seedling development were examined in this study. Five in each healthy and damaged trees of Pinus thunbergii growing in air polluted area for 12 years were chosen based on visible foliar injury and growth. The cones of P. thunbergii, which were collected from healthy and damaged trees, were analyzed for physical characteristics and seeds from the cones were used to test germination percentage under $O_3$ treatment. The germinated seeds were continuously exposed to $O_3$ treatment and the lipid peroxidation and activities of antioxidative enzymes were determined for both seeds and seedlings. The $O_3$ treatment for seed germination and seedling development were conducted at three conditions: control, 150 ppb and 300 ppb of $O_3$. The non-treated seeds from the damaged trees showed 21.6% lower germination than those from the healthy ones. On the $O_3$ treatment of 300 ppb, seed germination decreased approximately 10% for the healthy trees and 19% for the damaged trees compared to that on the control. The seeds from the healthy trees showed significantly higher activities of superoxide dismutase (SOD), glutathione reductase (GR), and catalase (CAT) than those from the damaged trees. The activities of GR, ascorbate peroxidase (APX), and CAT decreased along with the increasing $O_3$ concentration in two tree grades. Malondialdehyde (MDA) content of seeds was not influenced by $O_3$ treatment for two tree grades. In seedling development, there were no significant differences for length and biomass of needle and root of two tree grades at both the control and 150 ppb of $O_3$. At 300 ppb of $O_3$ treatment, however, the length and biomass of needle and stem decreased for two tree grades but no significant differences was detected in root. The seedlings from the damaged trees were more sensitive to the $O_3$ treatment, showing higher activities of SOD, APX, and CAT and content of MDA compared to those from the healthy tree seedlings. Our results indicate that seed germination and seedling development are vulnerable to increasing $O_3$ concentrations and that attention must be paid to the individual selection of tree species for reforestation.