• 한국작물학회지
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• 제49권1호
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• pp.12-18
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• 2004

Activity of senescence-induced antioxidant enzymes in the detached rice seedlings (Oryza sativa L. cv. Dongjin) was examined. The levels of $\textrm{H}_2\textrm{O}_2$ content and peroxidase (POD) activity were gradually increased during leaf senescence, whereas catalase activity was decreased. The activity of superoxide dismutase (SOD) was increased, and ascorbate peroxidase (APX) and glutathione reductase (GR) were slightly increased until 3d and 4d of dark induced-senescence, and thereafter were decreased. The activation of all SOD isoforms showed a significant decrease after 6d and 7d. After 4d to 7d of dark senescence, there was a significant effect in enhancing the activity of APX-12 and -13 isoforms as compared with light, despite similar levels in total APX activity. GR-8 and -10 isoforms were more effective in leaf senescence at 4d to 7d, particularly with respect to dark-induced senescence. These results suggest that the metabolism of active oxygen species such as $\textrm{H}_2\textrm{O}_2$ is dependent on various functionally interrelated antioxidant enzymes such as catalase, peroxidase, SOD, APX and GR.

• 농약과학회지
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• 제11권2호
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• pp.72-81
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• 2007

일련의 새로운 O,O-dialkyl-1-phenoxyacetoxy-1-methylphosphonate (S) 유도체들의 반응성과 치환기가 변화함에 따른 오이(Cucumus Sativa)씨에 대한 발아전 제초활성과의 관계를 2D-QSAR 및 HQSAR 방법으로 검토하였다. 통계적으로 HQSAR 모델이 2D-QSAR 모델보다 양호하였으며 기질분자(S)와 PDH 효소중 $BH^+$ 이온(I) 사이의 경계분자궤도(FMO) 상호작용은 친전자성 반응이 우세하였다. 치환기의 효과로부터 기질분자 (S)내 $R_2$-치환기는 carbonyl 산소원자에 대한 친전자성 반응을, 그리고 phenyl 고리상 X,Y-치환기는 carbonyl 탄소원자에 대한 친핵성 반응에 기여하였으며 $R_2$-치환기보다 X,Y-치환기의 영향이 더 컸다. 2D-QSAR모델 (I 및 II)과 HQSAR 모델의 기여도로부터 X,Y-치환기의 길이가 길수록 제초활성이 증가하는 경향이었으며 적정한 ${\epsilon}LUMO$ 에너지($({\epsilon}LUMO)_{opt.}$=-0.479 e.v.)가 제초활성에 중요한 요소이었다. 그러므로 PDH 효소의 저해활성으로 인한 제초활성은 친핵성반응으로 진행될 것으로 예상되었다. 2D-QSAR 및 HQSAR 두 모델로부터 제초활성에 기여하는 기질분자(S)의 구조 특이성과 요소들을 새로운 제초제 설계에 적용할 수 있음을 시사하였다.

• 공업화학
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• 제34권3호
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• pp.299-306
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• 2023

본 연구에서는 무환자와 알팔파로부터 추출한 천연계면활성제를 유화제로 사용하여 반응표면분석법 중 중심합성계획모델(CCD-RSM)을 이용한 O/W 유화 제조공정의 최적화를 수행하였다. 공정변수로는 혼합유화제 첨가량, 천연유화제 혼합비율(무환자 사포닌/알팔파 사포닌), 유화시간으로 설정하였고 반응치로는 유화액의 유화안정화지수, 평균입자크기, 항산화능(DPPH 라디칼 소거활성)으로 설정하였다. 기초실험을 통한 계량인자범위는 혼합유화제 첨가량(12~14 wt%), 천연유화제 혼합비율(30~70%), 유화시간(20~30 min)으로 설정하여 CCD-RSM을 이용하여 최적화한 결과, O/W 유화 제조공정의 최적조건으로 혼합유화제 첨가량은 13.2 wt%, 천연유화제 혼합비율은 44.2%, 유화시간은 25.8 min으로 나타났으며, 이 조건에서 예측된 반응치로서 유화액의 유화안정도지수(ESI)는 88.7%, 평균입자크기(MDS)는 815.5 nm, 항산화능은 38.7%으로 계산되었다. 이를 실험을 통해 확인한 결과 유화액의 ESI는 90.6%, MDS는 830.2 nm, 항산화능은 39.6%으로 나타났으며 평균오차율은 2.1%이었다. 따라서 CCD-RSM을 실제 유화 제조에 적용하여 만족스러운 O/W 유화제조 공정조건을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1557-1567
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• 2012

Glutathione reductase (GR, E.C. 1.6.4.2) is an important enzyme that reduces glutathione disulfide (GSSG) to a sulfydryl form (GSH) in the presence of an NADPH-dependent system. This is a critical antioxidant mechanism. Owing to the significance of GR, this enzyme has been examined in a number of animals, plants, and microbes. We performed a study to evaluate the molecular properties of GR (OsGR) from rice (Oryza sativa). To determine whether heterologous expression of OsGR can reduce the deleterious effects of unfavorable abiotic conditions, we constructed a transgenic Saccharomyces cerevisiae strain expressing the GR gene cloned into the yeast expression vector p426GPD. OsGR expression was confirmed by a semiquantitative reverse transcriptase polymerase chain reaction (semiquantitative RT-PCR) assay, Western-blotting, and a test for enzyme activity. OsGR expression increased the ability of the yeast cells to adapt and recover from $H_2O_2$-induced oxidative stress and various stimuli including heat shock and exposure to menadione, heavy metals (iron, zinc, copper, and cadmium), sodium dodecyl sulfate (SDS), ethanol, and sulfuric acid. However, augmented OsGR expression did not affect the yeast fermentation capacity owing to reduction of OsGR by multiple factors produced during the fermentation process. These results suggest that ectopic OsGR expression conferred acquired tolerance by improving cellular homeostasis and resistance against different stresses in the genetically modified yeast strain, but did not affect fermentation ability.

• BMB Reports
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• 제40권3호
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• pp.412-418
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• 2007

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is an unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes. In the present work, the entire encoding region for Oryza sativa PHGPx was expressed in Escherichia coli M15, and the purified fusion protein showed a single band with 21.0 kD and pI = 8.5 on SDS- and IFE-PAGE, respectively. Judging from CD and fluorescence spectroscopy, this protein is considered to have a well-ordered structure with 12.2% $\alpha$-helix, 30.7%$\beta$-sheet, 18.5% $\delta$-turn, and 38.5% random coil. The optimum pH and temperature of the enzyme activity were pH 9.3 and 27$^{\circ}C$. The enzyme exhibited the highest affinity and catalytical efficiency to phospholipid hydroperoxide employing GSH or Trx as electron donor. Moreover, the protein displayed higher GSH-dependent activity towards t-Butyl-OOH and $H_2O_2$. These results show that OsPHGPx is an enzyme with broad specificity for hydroperoxide substrates and yielded significant insight into the physicochemical properties and the dynamics of OsPHGPx.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.23-23
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• 2017

Rice (Oryza sativa L.) is the world's most important cereal crop. In crop plant, chlorophyll content and leaf senescence could affect grain filling and yield. We analyzed a QTL associated with chlorophyll content and delayed leaf senescence using high chlorophyll near isogenic line (HC-NIL). HC-NIL derived from a cross between Oryza sativa cv. Hwaseong as a recurrent parent and wild species O. grandiglumis as a donor parent showed higher chlorophyll content than Hwaseong. To identify QTL associated with chlorophyll content, 58 $F_3$ and 38 $F_4$ lines were developed from a cross between HC-NIL and Hwaseong. For QTL analysis, simple sequence repeat (SSR) markers were used for genotyping and one-way ANOVA was conducted. A QTL for chlorophyll content (qCC2) was detected in chromosome 2 and explained 24.63% of phenotypic variation. The senescence effect of the qCC2 was examined in dark-induced incubation (DII). Detached leaves from Hwaseong and HC-NIL were incubated on 3mM MES buffer (pH 5.8) at $27^{\circ}C$ under complete dark condition. After 3 days of incubation, the Hwaseong leaves turned yellow, but the HC-NIL leaves were green. HC-NIL has higher chlorophyll content with delayed senescence than Hwaseong. These results indicated that qCC2 is associated with stay-green phenotype. To know whether the qCC2 is responsible for leaf functionality, ion leakage test and Fv/Fm measurement were performed. Both experiment results showed that differences were observed between Hwaseong and HC-NIL but it was not statistically significant. These results might suggest that the qCC2 is possibly related to chlorophyll content and non-functional stay-green phenotype.

• BMB Reports
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• 제38권4호
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• pp.432-439
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• 2005

$Ca^{2+}$/calmodulin transduction pathways have been implicated in mediating stress response and tolerance in plants. Here, three genes encoding calmodulin (Cam) members of the EF-hand family of $Ca^{2+}$-binding proteins were identified from Oryza sativa L. databases. Complementary DNA for each of the calmodulin genes, OsCam1, OsCam2, and OsCam3 were sequenced. OsCam1 and OsCam2 encode a conventional 148-amino acid calmodulin protein that contains four characteristic $Ca^{2+}$-binding motifs. OsCam3 encode a similar protein with a 38-amino-acid extension containing a putative prenylation site (CVIL) at the carboxyl terminus. RT-PCR showed that each of the genes is expressed in leaves and roots of 2-week old rice seedlings. By RNA gel blot analysis, OsCam1 mRNA levels strongly increased in response to NaCl, mannitol and wounding treatments. In contrast, OsCam2 mRNA levels were relatively unchanged under all conditions investigated. NaCl treatment and wounding also increased the OsCam3 mRNA level, but in a more transient manner. Our results indicate that although the expression of genes encoding different calmodulin isoforms is ubiquitous, they are differentially regulated by various stress signals. In addition, we have demonstrated that the calcium-channel blocker lanthanum chloride inhibited the induction of OsCam1 gene expression by both NaCl and mannitol treatments. These results suggest that osmotic stress induced expression of OsCam1 gene requires the $[Ca^{2+}]_{cyt}$ elevation that is known to occur in response to these stimuli.

• Natural Product Sciences
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• 제9권1호
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• pp.22-27
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• 2003

This study was designed to investigate the effects of main constituents of Nigella sativa (NS) seed on the survival and CNS responses in experimental animals. The toxicological investigations were conducted for the determination of median lethal doses $(LD_{50})$ of NS seed constituents [i.e. aqueous extract (AE), fixed oil (FO), volatile oil (VO)] and main components of its VO [i.e. thymoquinone (TQ), ${\alpha}-pinene$ (AP) and p-cymene (PC)]. A part of this study includes evaluation NS constituents in the induction of minimal neurological deficit (MND) as a parameter for neurotoxicity using chimney test. In this study, the i.p. $LD_{50}$ values of AE, FO, VO, TQ (suspended In 0.5%CMC), TQ (dissolved in corn oil), AP and PC, were 3020, 3371, 1853, 616.6, 90.3, 1726 and 1523 mg/kg, respectively. All the NS constituents can be considered moderately toxic ($LD_{50}$ ranged from 616.6 to 3371 mg/kg), except the oily solution of TQ, which was very toxic ($LD_{50}$ was 90.3 mg/kg). It appeared that the toxicity of the whole VO is mainly due to its content of TQ and to some extent PC. All the NS constituents induced different degrees of MND at certain dose levels. The median neurotoxic (or sedating) doses $(TD_{50})$ of AE, FO, VO, TQ (suspended in CMC) and AP and PC, were 950, 1403, 306, 88.1, >173 and 368 mg/kg, respectively. TQ was the most potent component in inducing MND, whereas the FO and AE were the least. Neurotoxicity induced by the VO in the chimney test may refer basically to its contents of TQ and to some extent PC and AP.

• Molecules and Cells
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• 제21권3호
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• pp.360-366
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• 2006

Up to 35% of the rice genome consists of various kinds of transposons, and CACTA and MITE are two of the major class 2 DNA transposons in the genome. We have employed the consensus sequences of Rim2/Hipa CACTA, Stowaway MITE Pangrangja, and Tourist MITE Ditto for transposon display (TD) analysis to locate them on a genetic map, with 58 SSR markers used to anchor them. The TD analysis produced a high profile of the polymorphisms between the parental lines, Oryza sativa var. Gihobyeo/O. sativa var. Milyang, in intraspecific $F_{15}$ RIL lines, locating 368 markers of Rim2/Hipa CACTA, 78 markers of Tourist MITE Ditto, and 22 markers of Stowaway MITE Pangrangja. In the segregation analysis, non-parental segregating bands and segregation distortion bands were observed. The recombinant genetic map spans 3023.9 cM, with 5.7 cM the average distance between markers. The TD markers were distributed unequally on the chromosomes because many TD markers were located in pericentric chromosomal regions except in the cases of chromosomes 2, 3, 6 and 9. Although the number of transposon markers was not sufficient to include all rice class 2 transposons, the current map of CACTA and MITE transposons should provide new insight into the genome organization of rice since no previous DNA transposon map is available.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2005년도 춘계학술발표회 요지
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• pp.32-33
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• 2005