• Plant Biotechnology Reports
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• 제2권2호
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• pp.113-122
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• 2008

Nitric oxide (NO) affects the growth and development of plants and also affects plant responses to various stresses. Because NO induces root differentiation, we examined whether or not it is involved in increased ROS generation. Treatments with sodium nitroprusside (SNP), an NO donor, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger, and $N{\omega}-nitro-{\text\tiny{L}}-arginine$ methyl ester hydrochloride (${\text\tiny{L}}-NAME$), an NO synthase (NOS) inhibitor, revealed that NO is involved in the adventitious root growth of mountain ginseng. Supply of an NO donor, SNP, activates NADPH oxidase activity, resulting in increased generation of $O_2{^{{\cdot}-}}$, which subsequently induces growth of adventitious roots. Moreover, treatment with diphenyliodonium chloride (DPI), an NADPH oxidase inhibitor, individually or with SNP, inhibited root growth, NADPH oxidase activity, and $O_2{^{{\cdot}-}}$ anion generation. Supply of the NO donor, SNP, did not induce any notable isoforms of enzymes; it did, however, increase the activity of pre-existing bands of NADPH oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase. Enhanced activity of antioxidant enzymes induced by SNP supply seems to be responsible for a low level of $H_2O_2$ in the adventitious roots of mountain ginseng. It was therefore concluded that NO-induced generation of $O_2{^{{\cdot}-}}$ by NADPH oxidase seems to have a role in adventitious root growth of mountain ginseng. The possible mechanism of NO involvement in $O_2{^{{\cdot}-}}$ generation through NADPH oxidase and subsequent root growth is discussed.

• 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.

• 한국축산식품학회지
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• 제26권1호
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• pp.15-19
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• 2006

The aim of this study was to determine the best processing conditions for producing of dried lean pork as a ready-to-serve product without using large-scale machines. Lean pork sausage was produced using 1.27% sodium chloride, 0.075% sodium polyphosphate, 0.06% sodium ascorbate, 0.075% sodium pyrophosphate, 0.009% sodium nitrite, 0.009% dextrin, 0.11% sodium glutamate and 1.4% spice mixture. The most appropriate slice thickness for drying was examined by slicing the sausage at a 0.5, 1 and 2 cm thickness. The drying temperatures were determined by drying the sausage slices at 35, 48 and $68^{\circ}$. The total drying period was for 12 hr, In order to examine the ability of this process to sterilize the pork, the raw meat materials were inoculated with Escherichia coli (E. coli). The optimal conditions for producing lean pork sausages were a 2 cm slice thickness and drying temperature of $68^{\circ}C$ for 12 hr, The moisture content water activity, color, hardness and pH were measured in the dried product. The product had a moisture content of 47.5% and a water activity of 0.93. There was a 47.7% percentage reduction in moisture. The dried product tested negative for E. coli even though the raw meat materials been inoculated with E. coli.

• Applied Biological Chemistry
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• 제37권2호
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• pp.105-109
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• 1994

꾸지 뽕나무의 줄기껍질로부터 P388 tumor cell line에 대한 세포 성장 저해활성과 흰쥐 간 microsome 분획의 과산화 지질을 사용하여 $Fe^{++}/\;ascorbate$법으로 측정한 결과 세포 성장 저해활성과 항산화활성을 갖는 5개의 flavonoid계 화합물을 분리하였다. 분리된 화합물의 구조분석을 실시한결과 각종 spectral data와 보고된 문헌에 의하여 taxifolin, orobol, eriodictyol, dihydrokaempferol, steppogenin으로 각각 동정되었다. 이 화합물들의 항산화활성은 $IC_{50}$이 각각 6, 3, 3, >50, $10\;{\mu}g/ml$이었고 taxifolin을 제외한 P388 cell line에 대한 세포 성장저해는 각각 $IC_{50}$이 0.18, 3.3, 15, $6.2\;{\mu}g/ml$이었다. 한편 Escherichia coli BE 1186, Salmonella thyphimurium SL 1102, Staphylococcus aureus IFO 12732, Staphylococcus aureus R 209, Candida albicans에 대해서는 항균활성을 나타내지 않았다.

• 한국작물학회지
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• 제52권3호
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• pp.264-273
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• 2007

In order to examine the cross-tolerance of two chilling-tolerant cultivars (Donganbyeo and Heukhyangbyeo) and two chilling-susceptible cultivars (Hyangmibyeo and Taekbaekbyeo) to salt, paraquat, and drought, changes of physiological response and antioxidant enzymes were investigated. The seedlings were grown in a growth chamber until the 4-leaf stage. The seedlings were exposed to chilling at $5^{\circ}C$ for 3 days. For drought treatment, the seedlings were subjected to drought by withholding water from plants for 5 days. For paraquat study, plants were sprayed with $300{\mu}M$ paraquat. For the salt stress, the seedlings were transferred to the Hoagland's nutrient solution containing 0.6% (w/v) NaCl for 4 days. Chilling-tolerant cultivars showed cross-tolerant to other stresses, salt, paraquat, and drought in physiological parameters, such as leaf injury, chlorophyll a fluorescence, and lipid peroxidation. The baseline levels of antioxidative enzyme activities, catalase (CAT) and peroxidase (POX) activities in chilling-tolerant cultivars were higher than in the chilling-susceptible cultivars. However, there were no differences in ascorbate peroxidase (APX) and glutathione reductase (GR) activities between chilling-tolerant and -susceptible cultivars in untreated control. CAT activity in chilling-tolerant cultivars was higher than that in chilling-susceptible cultivars during chilling, salt, and drought treatments, but not during paraquat treatment. However, other antioxidative enzymes, APX, POX, and GR activities showed no significant differences between chilling-tolerant and -susceptible cultivars during chilling, salt, paraquat, and drought treatments. Thus, it was assumed that CAT contribute to cross-tolerance mechanism of chilling, salt, and drought in rice 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

• 한국식품과학회지
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• 제19권6호
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• pp.506-514
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• 1987

밤(생율(生栗))으로 부터 peroxidase를 $(NH_4)_2SO_4$에 의한 염석 및 DEAE-cellulose column chromatography, sephadex G-100 column chromatography, HPLC방법으로 정제하였으며 정제도는 조효소에 비하여 65.3배였고, HPLC로 측정한 밤 peroxidase 분자량은 35,000으로 추정되었다. sephadexG-100 column chromatography 후 얻은 밤 peroxidase의 작용최적 pH는 5.0이었고, 작용최적 온도는 $50^{\circ}C$이었으며 $80^{\circ}C$에서 1.73분 열처리할 때 90%의 효소가 불활성화되었다. 본 효소는 OPDA 및 PPDA와 같은 방향족 amine류에 높은 활성을 나타내었다. OPDA와 $H_2O_2$에 대한 Km치는 각각 2.6mM과 10mM이었고, 저해작용은 L-ascorbic acid 와 sodium L-ascobate가 가장 컸으며, $Ca^{++}$과 $Ba^{++}$은 1mM과 5mM에서 현저히 효소활성을 증가시켰다.

• 한국자원식물학회지
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• 제19권6호
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• pp.649-654
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• 2006

The objective of present study was to investigate the anti oxidative and hepatoprotective effects of tomato extracts. Total antioxidant capacity and total antioxidant response were 5.5 and $19.8{\mu}g$ Trolox equivalent per mg of tomato extract, respectively. DPPH radical scavenging activity of tomato extracts ($10mg\;ml^{-1}$) was 70% as compared to 100% by pyrogallol solution as a reference. The effect of the tomato extracts on lipid peroxidation was examined using rat liver mitochondria induced by iron/ascorbate. Tomato extracts at the concentration of $0.5mg\;ml^{-1}$ significantly decreased TBARS concentration. Tomato extracts prevented lipid peroxidation in a dose-dependent manner. The effect of the tomato extracts on reactive oxygen species (ROS) generation was examined using cell-free system induced by $H_2O_2/FeSO_4$. Addition of $1mg\;ml^{-1}$ of tomato extracts significantly reduced dichlorofluorescein (DCF) fluorescence. Tomato extracts caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that tomato extracts significantly prevented ROS generation in vitro. The effect of tomato extracts on cell viability and proliferation was examined using hepatocyte culture. Primary cultures of rat hepatocytes were incubated with 1mM tert-butyl hydroperoxide (t-BHP) for 90 min in the presence or absence of tomato extracts. MTT values by addition of tomato extracts at the concentration of 2, 10, and $20mg\;ml^{-1}$ in the presence of t-BHP were 13, 33 and 48%, respectively, compared to 100% as control. Tomato extracts increased cell viability in a dose-dependent manner. These results demonstrate that tomato extracts suppressed lipid peroxidation and t-BHP-induced hepatotoxicity and scavenged ROS generation. Thus antioxidant and hepatoprotective effects of tomato extracts seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation.

• 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 Microbiology and Biotechnology
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• 제22권3호
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• pp.407-415
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• 2012

An endophytic bacterium, Bacillus thuringiensis GS1, was isolated from bracken (Pteridium aquilinum) and found to have maximal production of chitinase (4.3 units/ml) at 5 days after culture. This study investigated the ability of B. thuringiensis GS1 to induce resistance to Rhizoctonia solani KACC 40111 (RS) in cucumber plants. Chitinase activity was greatest in RS-treated plants at 4 days. ${\beta}$-1,3-Glucanase activity was highest in GS1-treated plants at 5 days. Guaiacol peroxidase (GPOD) activity increased continuously in all treated plants for 5 days. Ascorbate peroxidase (APX) activity in RS-treated plants was increased 1.5-fold compared with the control at 4 days. Polyphenol oxidase (PPO) activity in RS-treated plants was increased 1.5-fold compared with the control at 3 days. At 5 days after treatment, activity staining revealed three bands with chitinase activity (Ch1, Ch2, and Ch3) on SDS-PAGE of cucumber plants treated with GS1+RS, whereas only one band was observed for RS-treated plants (Ch2). One GPOD isozyme (Gp1) was also observed in response to treatment with RS and GS1+RS at 4 days. One APX band (Ap2) was present on the native-PAGE gel of the control, and GS1- and GS1+RS-treated plants at 1 day. PPO bands (Po1 and Po2) from RS- and GS1+RS-treated plants were stronger than in the control and GS1-treated plants upon native-PAGE at 5 days. Taken together, these results indicate that the induction of PR proteins and defense-related enzymes by B. thuringiensis GS1 might have suppressed the damping-off caused by R. solani KACC 40111 in cucumber plants.