• BMB Reports
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• 제32권4호
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• pp.385-392
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• 1999

The $NAD^+$-specific activity of a dual coenzyme-specific isocitrate dehydrogenase (IDH; EC 1.1.1.41) from the primitive fungus Pythium ultimum was investigated to elucidate the regulatory factors that may influence the intracellular distribution of carbon and the availability of intermediates, e.g. citrate, for fatty acid synthesis. Inhibition of $NAD^+$-IDH activity by diphospho- and triphosphonucleotides (ATP, ADP, and GTP) reflected the sensitivity of this enzyme to cellular energy charge even though monophosphonucleotides (AMP and GMP) had little effect on activity. NADPH, but not NADH, substantially inhibited $NAD^+$-IDH activity, showing noncompetitive inhibition with isocitrate. Oxalacetate and ${\alpha}$-ketoglutarate showed competitive inhibition with isocitrate, while citrate and cis-aconitate showed mixed-noncompetitive inhibition with isocitrate. Inhibition by these substances ranged from 29 to 46% at 10 mM. The inhibitory effect of oxalacetate was increased synergistically by glyoxylate, which alone caused 31% uncompetitive inhibition at 10 mM, and a mixture of the two substances at 1 mM each showed 98% inhibition of $NAD^+$-IDH activity. The regulation of $NAD^+$-IDH in Pythium ultimum seems to be a complex process involving mitochondrial metabolites. The addition of glyoxylate (3 mM) and oxalacetate (3 mM) to the culture medium resulted in the production of 49% more lipid by P. ultimum.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.135-135
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• 1998

To find an antagonist of Pythium ultimum, the causal agent of damping-off, numerous actinomycete strains were screened for in vitro inhibiting mycelial growth of the target fungus and producing bioactive metabolites. A strain identified as Streptomyces sp. G60655 was isolated and used for further antagonistic efficacy. The degree of antagonism between the fungus and G60655 was affected by the medium used. Furthermore, the preinoculation of the antagonist was found to be necessary to exhibit the maximum efficacy of antagonsim against the fungus. From the culture broth, a bioactive metabolite was detected and purified by solvent extraction, silica gel chromatography and preparative HPLC. The FAB-MS spectrum of the active compound showed a molecular ion peak at m/z 1101 (M + H)$\^$+/, suggesting the molecular weight of 1100. The UV absorptions at 242 and 323 nm indicated the presence of aromatic functions. The structure of this compound was identified as echinomycin, a depsipeptide antibiotic by spectroscopic studies including various NMR measurements. Echinomycin was inactive against several soil born fungi, but inhibited the mycelial growth of P. ultimum and its related oomycetous fungi.

• 한국유기농업학회지
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• 제24권3호
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• pp.465-484
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• 2016

본 연구는 자생식물인 순비기나무, 번행초, 사철쑥 등을 이용하여 친환경 농자재로 개발하기 위한 기초자료를 제공하고자 수용성 추출액 농도에 따른 Stemphylium vesicarium, Penicillium italicum, Sclerotinia sclerotiorum, Pythium ultimum, Botrytis cinerea, Rhizoctonia solani, Colletotrichum gloeosporioides 7종의 작물 병원균에 대해 생장을 조사하였다. 공여체식물별에 따른 수용성 추출액 농도가 증가됨에 따라 작물 병원균 균사의 생장이 감소하는 경항을 보이나 공여체식물과 병원균의 종류에 따라 억제의 정도차이를 보였다. Stemphylium vesicarium 경우 번행초와 사철쑥에서 농도 증가에 따라 각각 r = -0.857 (p<0.01), r = -0.868 (p<0.01)로 역의 상관을 보였고 Penicillium italicum은 순비기나무 r = -0.833 (p<0.01), Sclerotinia sclerotiorum은 순비기나무 r = -0.862 (p<0.01)와 사철쑥 r = -0.902 (p<0.01), Botrytis cinerea은 번행초 r = -0.896 (p<0.01)에서 역의 상관을 보였으며 Rhizoctonia solani과 Colletotrichum gloeosporioides은 모든 공여체식물에서 역의 상관을 보였다. 100% 처리구에서 균사의 생장억제 활성은 번행초인 경우 Pythium ultimum, Botrytis cinerea, Colletotrichum gloeosporioides에 대해 각각 94%, 50%, 80%의 억제율을 보였다. 사철쑥인 경우 Stemphylium vesicarium, Pythium ultimum에 대해 각각 43%, 90%의 억제율을 보였다. 황근은 Penicillium italicum, Sclerotinia sclerotiorum에서 각각 50%, 26%가 억제되었으며 무화과는 Rhizoctonia solani에 대해 74%가 균사 생장이 억제되었다. 항균활성을 보이는 수용체식물의 총 페놀 및 플라보노이드 함량은 사철쑥 16.15 mg/g, 무화과 7.81 mg/g, 순비기나무 6.18 mg/g, 황근 5.25 mg/g, 번행초 4.41 mg/g 순으로 조사되었고 플라보노이드는 사철쑥 27.57 mg/g, 순비기나무 12.49 mg/g, 무화과 11.45 mg/g, 황근 5.77 mg/g, 번행초 5.08 mg/g 순으로 분석되었다.

• The Plant Pathology Journal
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• 제15권2호
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• pp.112-118
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• 1999

Eleven species of Pythium were isolated from leaf blight symptoms on creeping bentgrass (Agrostis palustirs Huds.), Kentucky bluegrass (Poa pratenisis L.) and zoysiagrasses (Zoysia japonica Steud., and Z. matrella (L.) Merr.) planted on golf courses in Korea. Mycelial growth on potato carrot agar medium under various temperature conditions indicated that Pythium species obtained in this study could be divided into four groups based on their responses to temperature conditions. P. vanterpoolii was found to favor low temperature conditions with the optimum temperature of $25^{\circ}$, whereas P. aphanidermatum and P. myriotylum favored relatively high temperature conditions with the optimum temperature of $35^{\circ}$. Other species including P. arrhenomanes, P. catenulatum, P. graminicola, P. oligandrum, P. rostratum, P. torulosum, and P. ultimum were the intermediate group with the optimum temperature of 25~$35^{\circ}$. P. periplocum was similar to the intermediate group but the minimum temperature for its mycelial growth was $15^{\circ}$, which was approximately $5^{\circ}$ above that for the intermediate Pythium spp.group. In the pathogenicity tests conducted in the lab using potted plants, P. aphanidermatum, P.a arrhenomanes, P. catenulatum, P. graminicola, P. myriotylum, P. periplocum, P. rostratum, P. torulosum, P. ultimum, and P. vanterpoolii were found to be pathogenic to creeping bentgrass and Kentucky bluegrass. P. aphanidermatum, P. catenulatum, and P. graminicola were frequently isolated from leaf blight symptoms of creeping bentgrass and Kentucky bluegrass in golf courses during the warm and humid periods in July-August. On the other hand, P. vanterpoolii and P. torulosum were frequently isolated during the cool and humid periods in March-May, suggesting both species might be the major causes of leaf blight occurring in the spring time. Zoysiagrass was susceptible to P. arrhenomanes and the heterothallic Pythium sp. (Ht-F), showing stem and crown rot of turf-grasses at poorly drained areas under coool and humid or rainy conditions. P. oligandrum and the heterothallic Pythium sp. (Ht-L) isolated from creeping bentgrass were avirulent to all species of turfgrasses tested in this study.

• 생명과학회지
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• 제12권2호
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• pp.200-207
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• 2002

채소류 모잘록병균인 Pythium ultimun에 길항하는 미생물을 분리하기 위해 체소류 뿌리를 균원시료로 하여 약600여종의 근권미생물을 분리하였고, 그중 5종의 근권미생물이 길항력을 나타내어 가장 높은 길항력 높은 Yl-37 균주를 최종 선별하였다. 분리한 길항균 YJ-37을 Bergey's Mannual of Systematic Bacteriology의 방법에 준하여 미생물의 형태학적, 배양학적, 생리·생화학적 특성을 조사 하였으며, 미생물의 세포벽의 지방산의 조성으로 자동 동정하는 Sherlock system을 이용하는 방법과 16S rDNA의 부분염기서열을 결정하여 동정하는 방법 등 3가지 방법으로 조사한 결과, 근권 길항미생물인 YJ-37 균주는 Bacillus ehimensis로 동정되었다. 채소류 모잘록병균 Pythium ultimun과 Rhizoctonia solani AG-4 이외에도 B. ehimensis YJ-37에 의한 병원성 진균의 길항력을 조사한 결과, Alternaria altrata, Collectotrichum gloeosporioides, Didymella bryoniae, Fu-sarium moniliforme, F oxysporum, F. oxysporum cucumerinum, F. oxysporum niveum, Gloeosporium sp., Glomerella sp., G. cingulata, G. lagenaria, Penicillium digitatum, P. itazicum, Phytophthora capsici, Sclerotinia sclerotiorum, Stemprhyzium solani등 대부분의 병원균에 대해서 항진균 활성을 나타내었다.

• 한국식물병리학회지
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• 제11권3호
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• pp.252-257
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• 1995

Growth rates of the low temperature growing isolates, Pseudomonas fluorescens M45 and MC07, reached maximum stationary phase within 50 hrs at the low temperature, 4$^{\circ}C$. But an ordinary biocontrol agent P. putida Pf3 did not reach logarithmic growth phase until 80 hrs. The culture filtrates of M45 and MC07 significantly inhibited the mycelial growths of Pythium ultimum, Rhizoctonia solani and Phytophthora capsici. Detached cotyledons of cucumber grown on Murashige and Skoog agar medium were much enhanced in their growth, compared to those without the filtrates. Population densities of M45 and MC07 in the rhizosphere at 14$^{\circ}C$ were more stable than at 27$^{\circ}C$. When M45 and MC07 were treated into soil, the population density of MC07 continuously increased up to 9 days after treatment, and sustained the initial inoculum density up to 60 days. Cucumber damping-offs caused by P. ultimum and R. solani were significantly reduced by applying M45 as seed-inoculant and by soil treatment with MC07. The combined treatment of M45 and MC07 provided greater effect in reducing the disease incidence than that obtained by single treatments.

• Applied Biological Chemistry
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• 제39권1호
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• pp.20-24
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• 1996

근권토양에서 분리한 형광성 P. fluorescens ps88은 철이 결핍된 환경에서 2차대사산물인 siderophore라는 형광성물질을 분비하며, KMB배지에서 Pythium ultimum, Pyricularia oryzae, Rhizoctonia solani 및 Xanthomonas oryzae의 생육을 억제시켰다. 오이의 씨앗을 P. fluorescens ps88로 접종한 후 Fusarium의 발병율을 조사하였을 때 ps88 접종구에서는 이병율이 대조구에 비하여 50%가 감소하였다. Vermiculate에서 균을 접종한 후 오이의 생육촉진효과를 조사한 결과 대조구보다 현저한 생육증가가 관찰되었으며, MS배지에서 고추의 뿌리에 siderophore를 처리하였을 때 대조구에 비하여 미세한 뿌리의 발달이 관찰되었다.

• Journal of Applied Biological Chemistry
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• 제42권3호
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• pp.146-150
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• 1999

As a part of the integrated disease system in greenhouse, an antifungal fungus(AF1) was isolated from greenhouse soil. It exhibited strong inhibitory activites against Pythium ultimum, Phytophtora capsici, Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum based on dual culture on 1/5 strength of potato dextrose agar between antagonistic fungus and several plant pathogens. The antagonistic fungus was identified as Chaetomium cochliodes, based on morphological characteristics; the body of the perithecium bears straight or slightly wavy, unbranched hairs, whilst the apex bears a group of spirally coiled hairs. To investigate antagonistic principles, antifungal compound was extracted and fractionated by different solvent systems. An antifungal compound was isolated as pure crystal from is culture filtrate using organic solvent extraction and column chromatography, followed by preparative thin layer chromatography. The chemical structure of the purified antifungal compound was identified as chaetoglobosin A based on the data obtained form $^1H-NMR$, $^{13}C-NMR$, DEPT 90, 135, $^1H-^1H$ COSY, $^1H-^{13}C$ COSY and EI/MS. $ED_{50}$ values of the chaetoglobosin A against P. ultimum, P. capsici, R. solani, B. cinerea and F. oxysporum were 1.98, 4.01, 4.16, 2.67 and 35.14 ppm, respectively.

• 생명과학회지
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• 제12권4호
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• pp.416-422
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• 2002

B. ehimensis YJ-37은 생육촉진미생물이면서 모잘록병의 생물학적 방제균으로서 관찰되었다 길항균 B. ehimensis YJ-37의 무와 수박의 근권정착밀도는 인공용토에 존재하는 길항균의 평균밀도 $10^4$(cfu/g) 보다 주근과 측근에서 약 1.2~2배 이상의 증가하였다. 길항균 배양액으로 만든 토양미생물제제에 의한 무의 생육촉진은 토양미생물제제 무처리구에 비해 처리구에서 잎면적은 85% 자엽의 길이는 33% 뿌리의 길이는 23% 건조중량은 89% 촉진되었고, 수박의 경우도 무처리구에 비해 처리구에서 잎면적은 63% 자엽의 길이는 27% 뿌리의 길이는 25% 건조중량은 69% 촉진되었다. 토양미생물제제는 채소류 모잘록병균 R. solani AG-4와 P. ultimum 등의 생육을 억제하여 모잘록병을 유발시키지 못하게 하였으며, 생육촉진현상도 동시에 가져왔다.

• Applied Biological Chemistry
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• 제40권6호
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• pp.593-596
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• 1997

시설내 토양으로부터 식물병원균에 활성을 갖는 길항균을 탐색하는 과정에서 주요 식물 병원균인 Phytophthora capsici, Botrytis cinerea, Rhizoctonia solani, Pythium ultimum 및 Fusarium oxysporum에 활성을 갖는 4종류의 길항균(AF1, AE2, AE3, AF4)을 분리하였다. 이들 중 가장 높은 활성을 나타낸 AE2균을 동정한 결과, 생육한 균은 짙은 오렌지색과 황갈색을 띄었으며, 현미경의 morphology는 column형의 conidial head와 foot cell로 부터 곧게 뻗은 분생자와 2중층의 포자낭병을 형성했다. 그리고 MY20 agar 배지에서 투명한 둥근세포의 형성등이 Aspergillus terreus의 형태와 일치 하였다. Pot에서 병원균과 A. terreus를 동시에 처리하여 오이의 생육을 조사한 결과, 토양중량당 A. terreus의 균체를 1% 처리에 40%, 5% 처리에 57%, 10% 처리에는 75% 병발생율을 억제시켰다. 그리고 A. terreus의 배양여액으로부터 activity-guided fractionation을 실시하여 항진균성 물질인 화합물 I 을 얻었다. $^1H$, $^{13}C\;NMR$, DEPT, $^1H-^1H\;COSY$, HMQC, HMBC 및 질량스텍트럼을 분석한 결과 화합물 I 은 butyrolactone I (${\alpha}$-oxo-${\beta}$-(p-hydroxyphenyl)-${\gamma}$-(p-hydroxy-m-3,3-dimethyl-allylbenzyl)-${\gamma}$-methoxycarbonyl-${\gamma}$-butyrolactone, $C_{24}H_{24}O_7$, M.W. = 424)로 동정되었다.