• Mycobiology
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• 제46권1호
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• pp.33-46
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• 2018

Gray mold (Botrytis cinerea) is one of the most common diseases of strawberries (Fragaria${\times}$ananassa Duchesne) worldwide. Although many chemical fungicides are used for controlling the growth of B. cinerea, the risk of the fungus developing chemical resistance together with consumer demand for reducing the use of chemical fungicides have necessitated an alternative method to control this pathogen. Various naturally occurring microbes aggressively attack plant pathogens and benefit plants by suppressing diseases; these microbes are referred to as biocontrol agents. However, screening of potent biocontrol agents is essential for their further development and commercialization. In this study, 24 strains of yeast with antagonistic ability against gray mold were isolated, and the antifungal activity of the volatile and diffusible metabolites was evaluated. Putative mechanisms of action associated with the biocontrol capacity of yeast strains against B. cinerea were studied through in vitro and in vivo assays. The volatile organic compounds produced by the Galactomyces candidum JYC1146 could be useful in the biological control of plant pathogens and therefore are potential alternative fungicides with low environmental impact.

• Mycobiology
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• 제49권1호
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• pp.69-77
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• 2021

The application of antagonistic fungi for plant protection has attracted considerable interest because they may potentially replace the use of chemical pesticides. Antipathogenic activities confirmed in volatile organic compounds (VOCs) from microorganisms have potential to serve as biocontrol agents against pre- and post-harvest diseases. In the present study, we investigated Galactomyces fungi isolated from rotten leaves and the rhizosphere of cherry tomato (Lycopersicon esculentum var. cerasiforme). VOCs produced by Galactomyces fungi negatively affected the growth of phytopathogenic fungi and the survival of nematodes. Mycelial growths of all nine examined phytopathogenic fungi were inhibited on agar plate, although the inhibition was more intense in Athelia rolfsii JYC2163 and Cladosporium cladosporioides JYC2144 and relatively moderate in Fusarium sp. JYC2145. VOCs also efficiently suppressed the spore germination and mycelial growth of A. rolfsii JYC2163 on tomatoes. The soil nematode Caenorhabditis elegans exhibited higher mortality in 24 h in the presence of VOCs. These results suggest the broad-spectrum activity of Galactomyces fungi against various plant pathogens and the potential to use VOCs from Galactomyces as biocontrol agents.

• 한국균학회지
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• 제41권4호
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• pp.268-273
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• 2013

흰가루병균(Podosphaera fusca)에 의한 오이 흰가루병의 생물적 방제를 위하여 검정한 길항세균 B. amyloliquefaciens M27 균주는 4.0% 흰가루병이 발생하였고 무처리는 80.5% 발생하였다. MH와 LB배지에서 배양한 B. amyloliquefaciens M27 균주의 배양여액은 TSB, NB와 KB배지에서 배양한 배양여액보다 오이 흰가루병 방제효과가 우수하였다. M27 균주를 LB배지에서 배양한 배양여액 2, 5, 10, 20, 50배와 100배 처리한 경우 오이 흰가루병이 0%, 0%, 0%, 1.3%, 3.1%, 5.0%와 33.3%를 나타낸 반면에 무처리는 60.0%를 나타내었다. 7월, 10월과 12월에 오이 흰가루병이 발생할 때 LB배지에서 배양한 M27균주의 배양여액를 10배로 처리한 결과 88.9~98.9% 방제효과를 나타났다. 이와 같은 결과에서 B. amyloliquefaciens M27 균주의 배양여액은 오이 흰가루병에 방제에 매우 효과적이었다.

• 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.

• 농업생명과학연구
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• 제50권4호
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• pp.27-34
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• 2016

고추를 비롯한 채소작물에 심각한 피해를 야기하는 흰비단병의 생물학적 방제제를 개발하기 위하여 시설재배 토양에서 분리한 세균 중에서 흰비단병균인 Sclerotium rolfsii의 균사생장을 강하게 억제하는 균주를 선발하였다. 선발된 균주는 형태적, 생리적 분석과 16S rRNA 분석결과를 바탕으로 Bacillus amyloliquefaciens로 동정하였고 최종 선발균주를 Bacillus amyloliquefaciens KBC1009로 명명하였다. Bacillus amyloliquefaciens KBC1009의 배양특성을 조사한 결과 배양온도 30℃, pH 7의 조건에서 72시간동안 배양했을 때 배양 효율이 가장 우수하였으며, 배양에 적정한 영양원은 탄소원 glucose, 질소원 yeast extract를 사용했을 때 가장 높은 균수를 나타내었다. 포장에서 고추 흰비단병에 대한 Bacillus amyloliquefaciens KBC1009의 방제효과를 검정하기 위해 흰비단병원균을 토양에 접종하고 B. amyloliquefaciens KBC1009 배양액(5×108 cfu/ml)을 500배로 희석하여 10일 간격으로 3회 관주 처리한 결과 무처리대비 66.7%의 방제효과를 나타내었다.

• 한국균학회지
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• 제24권2호통권77호
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• pp.93-103
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• 1996

오이 덩굴쪼김병에 효과 있는 새로운 토양첨가제의 조제와 방제효과를 구명코저 본 연구를 1993년부터 지난 2년간 수행하였다. 석회를 포함한 14종의 무기성분(1%, w/w)을 성분별로 공시하여 억제효과를 조사하였는데 특히 $Al_2(SO_4)_3$, Alum 및 CaO 등의 Fusarium oxysporum f. sp. cucumerinum에 대한 포자발아 억제율은 72.8-97%였고 균사생장 억제율은 20.9-25%이었다. 한편 $Ca(NO_3)_2$는 균사생장에만, KCl, $K_2SO_4,\;NH_4NO_3$와 Urea는 포자발아에만 억제효과가 켰으며 그 억제율은 37.0-71.9%이었다. 결과적으로 7종의 무기성분이 선발되었다. 부숙소나무 수피등 2종의 유기물을 2, 5 및 10% 농도별로 공시한 결과 부숙소나무 수피는 알팔파 잎가루에 비하며 병원균의 포자발아나 균사신전 억제효과가 우수하였다. 소나무 수피와 무기성분(1%, w/w)을 혼합한 토양첨가제 처리토양에서 길항균(Tr-3)의 생장은 양호하였을 뿐만 아니라 병원군의 균사생장을 효과적으로 억제하였다. 그리고 길항세균(Pseudomonas sp.)여액에서의 병원균에 대한 길항력도 우수함이 확인되었다. 부숙 소나무수피와 알팔파 잎가루의 추출액배지에서의 균사생장 억제는 크지 않았으나 포자발아는 부숙 소나무수피액 처리에서 뚜렷하였고 농도에 따라 억제율이 90% 이상이었다. 무기성분과 부숙 소나무 수피가루 및 2종의 길항균을 토양에 혼합한 (1% w/w)구에서는 오이 덩굴쪼김병에 대한 방제효과가 폿트와 포장시험결과로 완벽함이 밝혀졌다. 따라서 토양 첨가제에 의한 오이 덩굴쪼김병의 생물학적 방제를 위하여 농가포장에서의 그 활용이 기대된다.

• The Plant Pathology Journal
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• 제26권4호
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• pp.386-393
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• 2010

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.

• 한국균학회지
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• 제26권4호통권87호
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• pp.458-465
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• 1998

사과탄저병균, Glomerella cingulata에 선택적으로 항균활성이 있는 세균을 분리한 후 형태 및 생리화학적인 특성을 기준으로 동정하여 Pseudomonas sp. DGUM 5051로 명명하였다. 분리균의 생장을 위한 최적 pH와 온도는 각각 6.0과 $30^{\circ}C$이었고, 최적 항균활성을 나타내는 pH와 온도는 각각 7.0와 $24^{\circ}C$이었다. 사용한 복합배지 중 생장 및 항균활성에 있어서는 Brucella, Brain heart infusion과 LB배지가 우수하였다. Mineral salts 배지에 유일 탄소원, 질소원 및 인산원으로 각각 Sucrose, $KNO_3$ 및 $K_2HPO_4$를 첨가하였을 때 우수한 항균활성을 나타내었다. Pseudomonas sp. DGUM 5061의 항균활성은 배양 12시간 경과 후에 나타났다.

• Journal of Microbiology
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• 제43권3호
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• pp.308-312
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• 2005

The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeosporioides at the late exponential stage. The transcription profiles of the B. lentimorhus WJ5, both with and without a challenge from C. gloeosporioides, were analyzed using custom DNA chips containing 2,000 genome fragments. A total of 337 genes were expressed, with 87 and 47 up- and down-regulated, respectively. Of these, 12 genes, which were involved in central carbon metabolisms, and 7 from minor catabolism were relatively highly up-regulated (> 10 fold) and down-regulated (< 0.2 fold), respectively. Nine genes, which were thought to be related to the antifungal activity, were also up-regulated, but their levels were not so high (2.0 - 9.7 folds). From the results, during the early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporioides, nutrient competition seemed to occur; therefore, the genes from central carbon metabolisms could be up-regulated, while those from minor catabolism could be down-regulated.

• Journal of Forest and Environmental Science
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• 제33권4호
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• pp.342-347
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• 2017

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.