• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2005년도 국제학술심포지움 The 44th Annual Meeting of Korean Society for Life Science
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• pp.23-36
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• 2005

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed In yeast. To investigate whether .Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various orgarusms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs In detail. PBIl is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Bax-induced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower lovels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. H$_{2O2}$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of H2O2 treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased In the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 i'n vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation In situ. Thus, AtNDPK2 appears to play a novel regulatory role in H2O2-mediated MAPK signaling in plants.

• 생명과학회지
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• 제15권3호
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• pp.382-386
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• 2005

산성비가 식물 생장에 미치는 영향과 식물의 생화학적 방어반응을 조사하고자 봉선화를 이용하여 인공 산성비(pH 2.0, 3.0, 4.0, 5.6) 실험을 수행하였다. 산성비의 pH가 낮을수록 생육 피해는 심하게 나타났으며 pH 3.0 이하의 처리에 의해 잎에 암회색 또는 적갈색의 괴사반점이 생성되었다. MDA 함량은 pH 2.0 처리에서 약 $40\%$의 증가를 나타내었다. 산성비의 $H^+$ 부하량 증가에 따라 산화형인 DHA 및 GSSG의 함량이 증가하였다. 항산화효소인 SOD, APX, DHAR, GP등의 활성도 산성비의 $H^+$ 부하량의 증가에 따라 크게 증가하는 것으로 나타났다. 이상의 결과로 볼 때 산성비는 봉선화 식물에 활성산소 생성에 의한 산화스트레스를 일으키며, 이를 무독화하기 위해 식물의 생화학적 방어반응이 작용하는 것으로 사료된다

• 한국잡초학회지
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• 제17권4호
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• pp.431-438
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• 1997

유해(有害)한 활성산소종(活性酸素種)들을 생성하는 환경 stress에 대한 식물의 적응과정(適應過程)을 파악하기 위한 일환으로서, 호박을 식물재료로 공시하고 Mg이 결핍(缺乏)된 영양액(營養液)에서 재배하여 산화적(酸化的) stress 조건을 부여하였을 때, 나타나는 과산화(過酸化)의 피해정도와 항산화효소(抗酸化酵素)들의 활성변동(活性變動)을 조사하였다. 충분한 양의 Mg이 공급된 영양액에서 재배된 호박의 잎에서는 Mg, 엽록소(葉綠素), 단백질(蛋白質) 함량(含量) 및 항산화효소(抗酸化酵素)들의 활성(活性)이 다소 증가 되었거나 거의 일정하게 유지되었다. 그러나 Mg이 결핍(缺乏)된 영양액(營養液)에서 재배된 호박의 잎에서는 Mg, 엽록소(葉綠素), 단백질(蛋白質) 함량(含量)은 현저히 감소된 반면, 항산화효소(抗酸化酵素)인 AP, GR 및 SOD의 활성(活性)이 크게 증가되었다. 본 연구에서 얻어진 결과들은 식물에서 Mg의 결핍(缺乏)으로 이하여 해로운 활성산소종(活性酸素種)이 생성되며 동시에 이들 활성산소종(活性酸素種)의 유해(有害)한 작용을 소거(消去)하기 위한 항산화효소(抗酸化酵素)의 활성(活性)도 유도(誘導)되는 것을 시사한다.

• Journal of Plant Biotechnology
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• 제37권4호
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• pp.529-538
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• 2010

Proline accumulates in plants under environmental stresses including saline stress and alkaline stress. Here, we investigated the responses to two different stresses, saline stress (200 mM NaCl) and alkaline stress (100 mM $Na_2CO_3$) in two Leymus chinensis (Trin.) genotypes, LcWT07 and LcJS0107, and effects of exogenous proline on the activities of antioxidant enzymes. Both saline stress and alkaline stress significantly induced the accumulation of proline in leaves of the two genotypes after 96 h, and alkaline stress caused a transient and significant increase in LcJS0107 plants at 6 h. A reduction in the activities of catalase (CAT, EC 1.11.1.6) and ascorbate peroxidase (APX, EC 1.11.1.11), but not in the activity of superoxide dismutase (SOD, EC 1.15.1.1), was detected in plants exposed to saline and alkaline stresses. Remarkable decrease in relative water contents (RWC) was found in 144 h stressed plants. However, lipid peroxidation estimated by malonyldialdehyde (MDA) content in leaves remained relatively stable. With the addition of exogenous proline, it did not cause changes of proline levels in two genotypes, but combined with saline or alkaline stress, the exogenous application of proline significantly induced proline accumulation after even short treatment periods. Combined with salt stress, the exogenous application also increased the activities of CAT and APX. These results indicated that exogenous proline not only increases proline levels in vivo as a osmotic adjustment under stress, but mitigates the detrimental effects of saline and alkaline stresses by increasing the activities of antioxidant enzymes.

• Journal of Plant Biotechnology
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• 제50권
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• pp.1-10
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• 2023

고구마(Ipomoea batatas L. Lam)는 영양소, 가공 식품, 동물 사료 및 색소 재료의 유용한 공급원으로 이용 가능한 경제적으로 중요한 대표적인 뿌리 작물이다. 일반적으로 고구마의 저장 뿌리는 수확 후 저장 기간 동안 다양한 미생물과 질병에 의한 부패에 노출되기 쉽다. 수확 후 큐어링은 저장기간 동안 상처를 치유하고 미생물에 의한 부패를 방지하기 위한 가장 적합한 수단으로 알려져 있다. 본 연구에서는 큐어링과 연관된 분자적 기작에 관여하는 단백질들을 확인하기 위해, 큐어링 처리 후 저장기간 동안 단백질체의 변화를 분석하였다. 33℃ (큐어링) 및 15℃ (대조군)에서 3일 동안 처리하고 8주의 저장 기간이 지난 후 2D 전기영동 분석을 통해 단백질 spot의 변화를 확인한 결과, 31개 단백질 spot의 발현량이 차이 나는 것을 확인하였으며, 이들 중 15개의 단백질 spot을 동정하여 그 특성을 분석하였다. 동정된 단백질 중 alphaamylase (spot 1)는 큐어링 처리구에서만 발현량이 증가하였으며, probable aldo-keto reductase 2-like (spot 3) 및 hypothetical protein CHGG_01724 (spot 4)는 큐어링 및 대조구에서 동시에 발현량이 증가하였으나, sporamin A (spot 10)는 큐어링 및 대조구에서 발현량이 감소하였다. 한편, 대조구에서 enolase (spot 14)는 발현량이 증가하였으나, chain A of actinidin-E-64 complex+ (spot 19), ascorbate peroxidase (spot 22) 및 여러 sporamin 단백질들(spot 20, 21, 23, 24, 27, 29, 30 및 31)은 발현량이 감소하였다. 본 연구의 결과는 고구마 저장 뿌리에서 큐어링 처리와 관련된 단백질의 동정 및 수확 후 저장 기간동안 병 저항성과 관련된 기작에 대한 이해를 높이며, 향후 저온 저장 능력이 향상된 신품종 개발을 위한 후보 유전자의 도출에도 기여할 수 있을 것이다.

• Journal of Plant Biotechnology
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• 제5권3호
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• pp.143-148
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• 한국산림과학회지
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• 제96권5호
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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 $O_3$.

• 한국산림과학회지
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• 제97권5호
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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.

• 한국농림기상학회지
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• 제11권1호
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• pp.3-12
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• 2009

수목의 오존 내성을 단기 노출 후 나타난 생리적 피해와 생화학적 반응들의 표준화 지수로 결정하였다. 모감주나무, 벽오동, 때죽나무, 물푸레나무, 백당나무를 150ppb 오존 노출 시킨 후, 광합성 특성, MDA 함량 및 항산화효소의 활성을 측정하여 피해지수는 광합성 파라미터와 MDA 함량 변화를 이용하여 계산하였으며, 내성지수는 항산화효소의 활성 변화를 측정하여 산출하였다. 순양자수율, 탄소고정효율, 광호흡속도는 오존 노출시간에 따라 감소하여, 스트레스 평가지표로 적당한 것으로 판단되었다. 항산화효소의 활성은 수종, 노출시간 및 효소 종류에 따라 다양한 결과를 보여 주었다. 모감주나무 SOD 활성은 오존 노출시간에 따라 증가하였고, 물푸레나무의 SOD 활성은 오존 노출 6 시간 후에 증가하였으며, 벽오동의 CAT 활성은 무처리보다 낮았다. 표준화 지수를 기초로 한 5개 수목의 오존 내성은 두 내성 수종(물푸레나무>모감주나무)과 세 민감성 수종(때죽나무>벽오동>백당나무)으로 구분되었다.

• 한국환경과학회지
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• 제19권12호
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• pp.1323-1334
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• 2010

The effect of nitric oxide (NO) on antioxidant system and protective mechanism against oxidative stress under UV-B radiation was investigated in leaves of maize (Zea mays L.) seedlings during 3 days growth period. UV-B irradiation caused a decrease of leaf biomass including leaf length, width and weight during growth. Application of NO donor, sodium nitroprusside (SNP), significantly alleviated UV-B stress induced growth suppression. NO donor permitted the survival of more green leaf tissue preventing chlorophyll content reduction and of higher quantum yield for photosystem II than in non-treated controls under UV-B stress, suggesting that NO has protective effect on chloroplast membrane in maize leaves. Flavonoids and anthocyanin, UV-B absorbing compounds, were significantly accumulated in the maize leaves upon UV-B exposure. Moreover, the increase of these compounds was intensified in the NO treated seedlings. UV-B treatment resulted in lipid peroxidation and induced accumulation of hydrogen peroxide ($H_2O_2$) in maize leaves, while NO donor prevented UV-B induced increase in the contents of malondialdehyde (MDA) and $H_2O_2$. These results demonstrate that NO serves as antioxidant agent able to scavenge $H_2O_2$ to protect plant cells from oxidative damage. The activities of two antioxidant enzymes that scavenge reactive oxygen species, catalase (CAT) and ascorbate peroxidase (APX) in maize leaves in the presence of NO donor under UV-B stress were higher than those under UV-B stress alone. Application of 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3- oxide (PTIO), a specific NO scavenger, to the maize leaves arrested NO donor mediated protective effect on leaf growth, photosynthetic pigment and free radical scavenging activity. However, PTIO had little effect on maize leaves under UV-B stress compared with that of UV-B stress alone. $N^{\omega}$-nitro-L-arginine (LNNA), an inhibitor of nitric oxide synthase (NOS), significantly increased $H_2O_2$ and MDA accumulation and decreased antioxidant enzyme activities in maize leaves under UV-B stress. This demonstrates that NOS inhibitor LNNA has opposite effects on oxidative resistance. From these results it is suggested that NO might act as a signal in activating active oxygen scavenging system that protects plants from oxidative stress induced by UV-B radiation and thus confer UV-B tolerance.