헤테로폴리산 담지 메조포러스 물질에서 아이소프로필 나프탈렌의 알킬화반응

김명완,김왕기,김종호,서곤 전남대학교 촉매연구소 2000 觸媒硏究 論文集 Vol.21 No.-

The alkylation of 2-isopropyl naphthalene with isopropyl alcohol was studied over heteropoly acid catalyst supported on mesoporous material. The physicochemical state of loaded heteropoly acid was investigated using XRD, nitrogen adsorption and FT-IR techniques. Heteropoly acid was highly dispersed on the wall of mesoporous material, and retained its Br□nsted acidity. The initial conversion of 2-isopropyl naphthalene was high over the heteropoly acid catalyst supported on mesoporous material, while that was negligible over bulk heteropoly acid. The high selectivity for β,β - diisopropyl naphthalene was explained by the uniform mesopore inducing transition-state shape selectivity.

Overexpression of rice isoflavone reductase-like gene (<i>OsIRL</i>) confers tolerance to reactive oxygen species

Kim, Sang Gon,Kim, Sun Tae,Wang, Yiming,Kim, Sung-Kun,Lee, Chang Hoon,Kim, Keun-Ki,Kim, Ju-Kon,Lee, Sang Yeol,Kang, Kyu Young Blackwell Publishing Ltd 2010 Physiologia plantarum Vol.138 No.1

<P>Isoflavone reductase is an enzyme involved in isoflavonoid biosynthesis in plants. However, rice isoflavone reductase-like gene (<I>OsIRL,</I> accession no. AY071920) has not been unraveled so far. Here, we have characterized its behavior in response to oxidizing agents. Using Northern and Western blot analyses, the <I>OsIRL</I> gene and protein were shown to be down-regulated in young seedling roots treated with reduced glutathione (GSH) and diphenyleneiodonium (DPI), known quenchers of reactive oxygen species (ROS). The <I>OsIRL</I> transcript level in rice suspension-cultured cells was also found to be induced by oxidants such as hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>), ferric chloride (FeCl<SUB>3</SUB>), methyl viologen (MV) and glucose/glucose oxidase (G/GO), but down-regulated when co-treated with GSH. Furthermore, to investigate whether overexpression of <I>OsIRL</I> in transgenic rice plants promotes resistance to ROS, we generated transgenic rice lines overexpressing the <I>OsIRL</I> gene under an abscisic acid (ABA) inducible promoter. Results showed that the <I>OsIRL</I> transgenic rice line activated by ABA treatment was tolerant against MV and G/GO-induced stress in rice leave and suspension-cultured cells. Our results strongly suggest the involvement of <I>OsIRL</I> in homeostasis of ROS.</P>

Effect of Phytohormones and Chemical Inhibitors on Pathogenesis-related Genes Identified by Differential Hybridization in Rice Suspension Culture Cells

Kim, Sang-Gon,Wu, Jing-Ni,Wang, Yiming,White, Ethan E.,Choi, Young-Whan,Kim, Keun-Ki,Choi, In-Soo,Kim, Yong-Cheol,Kim, Sun-Hyung,Kang, Kyu-Young,Kim, Sun-Tae The Korean Society of Plant Pathology 2010 Plant Pathology Journal Vol.26 No.4

In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

In-depth insight into in-planta apoplastic secretome from rice and Cochliobolus miyabeanus during their interaction

U G Kim,Soon Jae Kwon,Yiming Wang,Sang Gon Kim,Kyu Young Kang,Sun Tae Kim 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

Here, we first demonstrate that identification of rice brown spot disease fungus (Cochliobolus miyabeanus, C. miyabeanus) proteins is possible in infected tissues using in planta apoplastic proteome with non-destructive tissues. In planta apoplastic proteins from rice leaves inoculated with C. miyabeanus, CM2 (compatible race), were isolated by vacuum infiltration with CaCl2/Na-acetateextractionbuffer, separated by SDS-PAGE, and identified by MudPIT. Of the 529 proteins that were identified by MudPIT, a large proportion (490) was from the rice. Numerous carbohydrate metabolic process (48), oxidation and reduction (44), response to oxidative stress (20%) were identified and confirmed their expression at RNA levels using microarray. Bioinformatic analysis showed that 176 and 39 of these proteins have a signal peptide in rice and rice brown spot fungus, respectively, using Signal P. The large proportion of proteins interestingly identified from the in planta apoplast were involved inprotease, hydrophobin, and host cell wall hydrolysis (Xylanase, beta-glucosidase) derived from pathogen. Thus, we suggest that in planta rice apoplastic secretome will be an important clue to understand the rice-rice brown spot fungus interactions.

A Comparative Proteomics Survey of Proteins Responsive to Phosphorous Starvation in Roots of Hydroponically-grown Rice Seedlings

Kim, Sang-Gon,Wang, Yiming,Lee, Chang-Hoon,Mun, Bong-Gyu,Kim, Pil-Joo,Lee, Sang-Yeol,Kim, Yong-Chul,Kang, Kyu-Young,Rakwal, Randeep,Agrawal, Ganesh Kumar,Kim, Sun-Tae The Korean Society for Applied Biological Chemistr 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.5

Rice takes up phosphorous (P) as major nutrient source for its growth and development when grown under anaerobic water-logged soil conditions. To better understand the underlying mechanisms and to develop potential protein biomarkers of P-starvation, hydroponically-grown rice seedlings in the complete media and phosphorus absence (P-starvation) of phosphorous nutrient solutions were investigated for physiological and proteome changes. The P-starvation manifested significant reduction in root growth in three-week-old seedlings compared to respective complete media. Furthermore, P-starvation also showed increased activity of acid phosphatase in roots of one- and three-week-old seedlings, suggesting that experimental design is suitable for proteomics survey of P-starvation responsive proteins. Two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis of total root protein from three-week-old seedlings identified 10 P-starvation responsive protein spots out of 140 high-quality protein spots. Identified 10 proteins were involved in metabolism and defense/stress response. Out of 10, 2 and 8 protein spots were found to be up- and down-regulated, respectively. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis of corresponding genes of four randomly selected proteins, including putative glyceraldehydes-3-phophate dehydrogenase (G3PDH, spot R1), S-adenosyl-L-methionine synthetase (SAMS, spot R4), ATP synthase subunit alpha (spot R6), and root-specific pathogenesis-related protein 10 (PR-10, spot R8), showed that just as protein abundance, these proteins are also regulated at the transcript level. Results suggest identified P-starvation responsive proteins are involved in maintaining nutrient homeostasis and/or associated with changes in root physiology under the absence of P.

Development of Parameters for Diagnosing Laryngeal Diseases

Kim, Yong Ju,Wang, Soo Geun,Kim, Gi Ryun,Kwon, Soon Bok,Jeon, Kye Rok,Back, Moo Jin,Yang, Byung Gon,Jo, Cheol Woo,Kim, Hyung Soon 한국음성과학회 2003 음성과학 Vol.10 No.1

Many people suffer from various laryngeal diseases. Since we can notice voice change easily, acoustic analysis can be helpful to diagnose the diseases. Several attempts have been made to clarify the relation between the parameters and the state of ssick vocal folds but any decisive parameters are not found yet. The purpose of this study was to select and develop those parameters useful for diagnosing and differentiating laryngeal diseases. We examined eight MDVP parameters, and two additional MFCC and LPC parameters obt ained from the production of an open vowel by 252 subjects with or without laryngeal diseases. Using a statistical procedure through the artificial neural networks, we attempted to differentiate laryngeal disease groups. Results showed that the LPC parameters indicated the highest differentiating rate by the networks followed by the MFCC and the MDVP parameters. In addition, Jita, Shim and NHR among the MDVP parameters came out better parameters in diagnosing laryngeal diseases.

Differentially expressed proteins in the liver of Gulo-/- mice following treatments with Helicobacter pylori and diethylnitrosamine

Gon-Sup Kim , Arulkumar Nagappan, Hyeon-Soo Park, Kwang-Il Park, Jin-A Kim, Gyeong-Eun Hong, Silvia Yumnam, Eun-Hee Kim, Won-Sup Lee, Wang-Jae Lee, Myung-Je Cho, Woo-Kon Lee, Chung-Kil Won 충북대학교 동물의학연구소 2013 Journal of Biomedical and Translational Research Vol.14 No.2

Vitamin C (ascorbic acid) is an essential nutrient of most living tissues. We established a strain of Gulo-/- mice with known deficiency, in which vitamin C intake can be controlled by diet, like humans, and investigated the differentially expressed proteins following treatments with Helicobacter pylori and diethylnitrosamine (DENA) in the liver of Gulo-/- mice using a proteomic approach. Expression of p53, 14-3-3ε and 14-3-3δ in Gulo-/- mice liver tissue was analyzed by immunohistochemistry. 2-DE maps constructed from Gulo-/- mice liver and differentially expressed proteins in liver tissue were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF/MS). In Gulo-/- mice after H. Pylori infection, followed by treatment with DENA, no differences in p53, 14-3-3ε and 14-3-3δ were observed by immunohistochemistry. Proteome analyses using MALDI-TOF/MS resulted in successful identification of 12 proteins (nine proteins were up-regulated and three were down-regulated). Specifically, peroxiredoxin-6 and Alpha-1-antitrypsin 1-4 were up-regulated in liver after H. Pylori infection followed by treatment with DENA. These results indicated that oral supplementation with vitamin C led to rescue of Gulo-/- mice from vitamin deficiency, and protected the liver from H.pylori infection and/or DENA effect, and vitamin C also protected the liver against oxidative stress

Proteasome Inhibitors Affect Appressorium Formation and Pathogenicity of the Rice Blast Fungus, Magnaporthe oryzae

Wang, Yiming,Kim, Sang-Gon,Wu, Jingni,Yu, Seok,Kang, Kyu-Young,Kim, Sun-Tae The Korean Society of Plant Pathology 2011 Plant Pathology Journal Vol.27 No.3

Previously, we identified the 20S proteasome ${\alpha}$-subunit of Magnaporthe oryzae (M. oryzae) induced during appressorium formation, and detected an increase in multiple protein ubiquitination during the early appressorium formation process (Kim et al., 2004). In this study, we further attempted to determine whether the proteasome is involved in the appressorium formation of M. oryzae both in vitro and in planta, using proteasome inhibitors. A significant increase in 20S proteasome during fungal germination and appressorium formation was observed using Western blot analysis with 20S proteasome antibody, demonstrating that proteasome-mediated protein degradation was involved in appressorium formation. Pharmacological analysis using proteasome inhibitors, MG-132, proteasome inhibitor I (PI) and proteasome inhibitor II (PII) revealed that germination and appressorium formation were delayed for 4 to 6 h on rice leaf wax-coated plates. Similarly, the treatment of proteasome inhibitors with fungal conidia on the rice leaf surface delayed appressorium formation and host infection processes as well. Additionally, fungal pathogenicity was strongly reduced at 4 days' postfungal infection. These data indicated that the fungal 20S proteasome might be involved in the pathogenicity of M. oryzae by the suppression of germination and appressorium formation.

Physiological and Proteomics Analysis to Potassium Starvation in Rice

Kim, Sang-Gon,Wang, Yiming,Lee, Chang-Hoon,Chi, Yong-Hun,Kim, Keun-Ki,Choi, In-Soo,Kim, Yong-Chul,Kang, Kyu-Young,Kim, Sun-Tae The Korean Society of Environmental Agriculture 2011 한국환경농학회지 Vol.30 No.4

BACKGROUND: Potassium (K) is one of the macronutrients which are essential for plant growth and development. Its deficiency in paddy soils is becoming one of the limiting factors for increasing rice yield in Asia. METHODS AND RESULTS: To investigate physiological symptoms under K-starvation (NP) compared with complete media (NPK) condition, we measured shoot/root length, weight, nutrients, and patterns of protein expression. The shoot growth was significantly reduced, but root growth was not affected by K-starvation. However, biomasses were decreased in both shoot and root. Uptake of K was reduced up to 85%, while total concentrations of P, Ca, Mg, Na were increased in root and shoot. To better understand the starved K mechanism of rice, comparative proteome analysis for proteins isolated from rice leaves was conducted using 2-DGE. Five spots of differentially expressed proteins were analyzed by MALDI-TOF MS. Analysis of these K-starvation response proteins suggested that they were involved in metabolism and defense. CONCLUSION(s): Physiological and 2-DGE based proteomics approach used in our study results in observation of morphology or nutrients change and identification of K-starvation responsive proteins in rice root. These results have important roles in maintaining nutrient homeostasis and would also be useful for further characterization of protein function in plant K nutrition.

Physiological and proteomic analysis of young rice leaves grown under nitrogen-starvation conditions

Kim, Sang-Gon,Wang, Yiming,Wu, Jingni,Kang, Kyu-Young,Kim, Sun-Tae The Korean Society of Plant Biotechnology 2011 Plant biotechnology reports Vol.5 No.4

Rice grown in anaerobic waterlogged soil accumulates ammonium as a major source of nitrogen (N). We have compared the physiological symptoms of rice seedlings subjected to N-starvation stress with those receiving sufficient N, based on measurements of shoot/root length and weight and an analysis of protein expression patterns. N starvation marginally increased root growth but notably decreased shoot biomass. N uptake was reduced by >50% in the roots and shoots of N-starved seedlings. To better understand the mechanism of N starvation in rice, we performed a comparative proteome analysis of proteins isolated from rice leaves. Twenty-five differentially expressed proteins were analyzed by matrixassisted laser desorption/ionization time-of-flight (TOF) mass spectrometry and electron spray ionization quadrupole TOF. Functional analysis of the N-starvation response proteins suggested their involvement in protein synthesis and fate, metabolism, and defense. These results indicate that these proteins may play important roles in regulating the plant's complex adaptation responses for N use during N starvation. The proteins may be useful for further characterization of protein function in plant N nutrition.