• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.142-146
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• 1996

옥수수 깜부기병균 Ustilago maydis 동포자를 풍건한 후 모래나 silica gel을 보존매체로 해서 4$^{\circ}C$나 그 이하 온도에서 24개월 두었을 때 동포자 생존율은 40~45%였다. 동포자는 28$^{\circ}C$ PDA상에서 쉽게 발아하여 많은 소생자를 형성하였다. U. maydis 동포자로부터 소생자 형성의 최적배지는 corn meal broth였고, carrot broth와 potato-dextrose broth의 순으로 좋았다. 소생자 형성에 가장 적합한 조건은 온도 28~3$0^{\circ}C$, pH5~7이었다. 소생자 형성은 암조건에서 보다 좋았다.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.205-212
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• 2011

Phenylalanine ammonia-lyase (PAL) from the maize pathogen Ustilago maydis was analysed immunologically to obtain insights into the structural relationships between plant PAL and fungal PAL and between PAL and histidine ammonia-lyase (HAL). Cross-reactivity was found among all the PAL proteins from different species tested, using antibodies raised against both plant and fungal PALs. Both anti-Alfalfa and anti-popular PAL antibodies strongly recognized plant PALs but only weakly recognized fungal PALs. Antibodies raised against U. maydis PAL only weakly recognized the Rhodotorula glutinis yeast PAL. The anti-U. maydis PAL antibodies showed low affinity for the plant PALs but they bound strongly to Pseudomonas bacterial HAL. Significant cross-reactivity between the two plant PAL antibodies and the bacterial HAL was also observed. Both the anti-Ustilago PAL and the anti-poplar PAL antibodies displayed similar enzyme inhibition patterns, including moderate inhibition of bacterial HAL activity. However, the bacterial HAL antibody inhibited only Ustilago PAL. The PAL and HAL antibodies tested showed no inhibition against yeast PAL. This is first report on the immunological relationships between PAL and HAL.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.249-253
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• 2011

Cetecol has been known as a component of melanin in teliospores of the corn smut fungus Ustilago maydis. Its metabolic precursor has been assumed to be benzoic acid but it has not been proven yet. This study was carried out to verify the synthesis of benzoic acid and to chase its metabolic origin in U. maydis. For this aim, the catabolic process of phenylalanine was investigated by culturing the fungus in the complete medium containing L-$^{14}C$-phenylalanine and $^{14}C$-trans-cinnamic acid. We detected trans-cinnamic acid, benzoic acid, 4-hydroxybenzoic acid and hydroxybenzoic acid derivatives from the extracts of the fungus cells and cultural filtrates by thin layered chromatography analysis. We also observed that the fungus could completely catabolize L-$^{14}C$-phenylalanine and produce $^{14}CO_2$ in the air. Conclusively, this study provided an evidence that U. maydis could produce benzoic acid through catabolic process of phenylalanine.

• Journal of Microbiology
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• v.35 no.4
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• pp.323-326
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• 1997

Toxin protein from Ustilago maydis virus SH14 isolated in Korea was purified using ethanol precipitation, cation exchange, gel filtration and anion exchange chromatography. The molecular weight of the purified protein was estimated to be 8.3 kDa by SDS-PAGE analysis. The Nterminal sequence of the protein is L-G-I-N-C(K)-R-G-S-S-Q--C(K)-G-L-S-G which is highly homologous with that of P4 toxin, but the amino acid composition and electrophoretic mobility in a native PAGE of the toxin protein were totally different from those of P4 toxin respectively. The SH14 toxin was shown to have immunological cross-reactivity about 50% with P4 toxin when examined by Western hybridization.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.184-188
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• 1993

Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.147-149
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• 1996

Ustilago maydis의 소생자는 물한천배지에서 접종후 8시간 후에 98% 이상 발아하였다. PDA를 포함한 기타 영야배지에서의 발아율도 이와 비슷하였다. 소생자 발아에 가장 적당한 온도는 3$0^{\circ}C$였으며 $25^{\circ}C$나 35$^{\circ}C$순으로 양호하였으나 2$0^{\circ}C$나 그 이하의 저온에서는 발아율이 급격히 떨어졌다. 소생자는 pH 5~9의 범위에서 80%이상 발아하였으나 최적 pH는 6~7이었다. 광조사는 소생자 발아에 큰 영향을 미치지 못하였다. 소생자는 자유수분(RH 100%)하에서 24시간 후 81%가 발아하였으나 상대습도가 93%나 그 이하에서는 발아율이 19~34%였다. 이 실험결과는 소생자 발아에 가장 큰 영향을 미치는 환경요인은 온도와 습도였으며 우리나라 기후조건에서 소생자는 7~8월의 한여름철 장마기간 동안 왕성하게 발아할 수 있음을 뜻하고 있다.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.60-60
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• 2015

Hybrid histidine kinase is a part of two-component system that is required for various stress responses and pathogenesis of pathogenic fungi. In the present study, Tco1, a homologue of human pathogen Cryptococcus neoformans Tco1 encoding a hybrid histidine kinase, was identified in corn smut pathogen Ustilago maydis by bioinformatic analysis. To explore the role of Tco1 in the virulence of U. maydis, mutants in which the tco1 gene was partially deleted were constructed by allelic exchange. The U. maydis tco1 mutants did show unaltered growth rate on axenic medium but were unable to produce conjugation tubes and develop fuzzy filaments, resulting in impaired mating of compatible strains. The expression levels of prf1, pra1, and mfa1 which are involved in the pheromone pathway significantly decreased in the tco1 mutants. In inoculation tests to host, the tco1 mutants showed significantly reduced ability in the production of anthocyanin pigments and tumor development on maize leaves. Overall, the combined results indicated that Tco1 plays important roles in sexual development and virulence of U. maydis by regulating the expression of the genes involved in the pheromone pathway.

• Korean Journal Plant Pathology
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• v.12 no.3
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• pp.291-296
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• 1996

옥수수 유묘접종시 깜부기병이 가장 잘 발병하는 조건은 4엽기 옥수수묘를 10\ulcorner~10\ulcorner/ml 농도의 소생자 현탁액으로 경엽살포한 후 28~3$0^{\circ}C$의 습실상에서 24시간 둔 후 꺼내어 온실에 두었을 때였다. 유묘의 엽기가 이보다 어리거나 접종원 농도가 이보다 높을 때, 혹은 습실상내 처리기간이 길어지면 접종후 유묘의 이른 고사를 초래하였다. 또한 습실상의 온도가 낮아질수록 깜부기병의 발병정도도 감소하였다. 깜부기병의 발생은 소생자 현탁액의 경엽살포 및 줄기주사접종에 의해서는 가능하였지만 토양관주에 의해서는 발병하지 않았다. 소생자 현탁액은 별도의 영양원의 첨가 없이도 발병을 잘 유도하였다. 동포자 현탁액을 직접 경엽살포하거나 줄기주사접종하여도 발병하였으나 소생자 현탁액접종에 비하여 발병정도가 현저히 낮았다.

• Journal of Microbiology
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• v.33 no.2
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• pp.154-159
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• 1995

SH-14, a novel killer strain of Ustilago maydis was isolated in Korea. It has been reported in other papers that the toxin specificity and double-stranded RNA pattern of SH-14 strain were different from other laboratory strains. In this paper, we analyzed the biochemical characteristics of U. maydis SH-14 virus. Three distinctive peaks were isolated from CsCl density gradient, designated as top (T), intermediate (I) and bottom (B) components. We found that the densities of each components, 1.285, 1.408 g/cm$\^$3/, respectively, are very similar to those of other strains. As previously reported by the analysis of dsRNA in each component, the dsRNA segments are separately encapsidated. Capsid protein of SH-14 virus consists of two proteins about 70 Kd shown by SDS-PAGE analysis. Electron microscopic examination of the virus particles revealed that UmV particles are very similar in size and morphology to all isolates as well as all lab-strains. In order to test immunological cross reactivity of UmV, werstern bolt analysis was carriedout with antiserum against A8 virus. All capsid protein had positive reaction against A8 antibody which indicated that UmV are immunologically cross-reactive with all isolates from Korea. The results presented in this paper may show that UmV isolated from SH-14 strain has very similar biochemical characteristics to those of other UmV. However, the difference in the toxin specificity and the molecular weight of toxin protein from the SH-14 strain has us to conclude that U. maydis SH-14 strain is a new killer type.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.1010-1022
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• 2017

Hybrid histidine kinase is part of a two-component system that is required for various stress responses and pathogenesis of pathogenic fungi. The Tco1 gene in human pathogen Cryptococcus neoformans encodes a hybrid histidine kinase and is important for pathogenesis. In this study, we identified a Tco1 homolog, UmTco1, in the maize pathogen Ustilago maydis by bioinformatics analysis. To explore the role of UmTco1 in the survival of U. maydis under environmental stresses and its pathogenesis, ${\Delta}umtco1$ mutants were constructed by allelic exchange. The growth of ${\Delta}umtco1$ mutants was significantly impaired when they were cultured under hyperosmotic stress. The ${\Delta}umtco1$ mutants exhibited increased resistance to antifungal agent fludioxonil. In particular, the ${\Delta}umtco1$ mutants were unable to produce cytokinesis or conjugation tubes, and to develop fuzzy filaments, resulting in impaired mating between compatible strains. The expression levels of Prf1, Pra1, and Mfa1, which are involved in the pheromone pathway, were significantly decreased in the ${\Delta}umtco1$ mutants. In inoculation tests to the host plant, the ${\Delta}umtco1$ mutants showed significantly reduced ability in the production of anthocyanin pigments and tumor development on maize leaves. Overall, the combined results indicated that UmTco1 plays important roles in the survival under hyperosmotic stress, and contributes to cytokinesis, sexual development, and virulence of U. maydis by regulating the expression of the genes involved in the pheromone pathway.