• The Plant Pathology Journal
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• 제16권4호
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• pp.189-199
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• 2000

To search for the antifungal substances, various actino-mycete isolates were obtained from various soils of Korea using plate dilution method on the humic acid vitamin agar plates. In the screening procedures using a dual culture method, 32 actionomycete isolates were selected, which showed the inhibitory activity against mycelial growth of plant pathogenic fungi Altirnaria mali, Colletotrichum gloeosporides, Fusarium oxysporum f.sp. cucumerinum, Magnaporthe grisea, Phytophthora capsici, and Rhizoctonia solani. Bioassay of the crude extracts from culture filtrates and mycelial mets revealed that 12 antagonistic actionomycetes produced highly active antifungal substances. Actinomycete strain S5-55 which showed the substantial antifungal activity against the tested fungi was selected for production of the antifungal substances. Based on the cytochemical and morphological characteristics, strain S5-55 was identified as a Streptomyces species. The results of the numerical identification using the TAXON program confirmed that Streptomyces strain S5-55 was identical with Streptomyces humidus including in TAXON major cluster 19. The production of antifungal substance was most favorable when S. humidus strain S5-55 was cultivated for 10 dats on soluble starch broth supplemented with $K_2$HPO$_4$. The antifungal substances active against the plant pathogenic fungi P. capsici and M. grisea were partially purified using $\textrm{C}_{18}$ reversed-phase column chromatography.

• 농약과학회지
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• 제2권1호
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• pp.40-45
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• 1998

온실 토양으로부터 주요 작물 병원균의 생육을 억제하는 길항곰팡이를 분리, 동정하였고, 그 균이 분비하는 향균물질을 분리하여 활성을 petriplate assay로 검정했다. 길항균의 foot cell, 분생포자 및 MY20 배지상의 균사 등 형태학적 특징에 근거하여 이 균을 Aspergillus terreus로 동정하였다. A. terreus의 배양여액을 chloroform으로 추출하고, 그 추출물을 column chromatography와 thin layer chromatography로 향균물질을 순수분리하였으며, 화학구조는 butyrolactone I이었다. 이 물질의 주요 작물병원균에 대한 $ED_{50}$은 Botrytis cinerea, Phytophthora capsici. Pythium ultimum, Rhizoctonia solani, 그리고 Fusarium oxysporum에 각각 9.7, 42.6, 23.3, 13.7, 102.7 ppm으로 나타났다.

• 한국환경농학회지
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• 제31권1호
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• pp.68-74
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• 2012

BACKGROUND: Some microorganisms extant in nature have ability to suppress various plant pathogens, and also can promote plant growth. Thus microorganisms are such great source of antimicrobial agents to develop antagonistic microorganism production and eco-friendly crop management. We isolated the microorganisms in various eco-friendly formulations. The suppressive abilities against plant pathogens have been characterized in vitro level. METHODS AND RESULTS: The indigenous microorganisms have been isolated from Cooked rice, Black sugar, Rice Bran, and Red clay using dilution plating method. Population of bacteria and fungi were above 107 in the all formulations. We isolated and pure cultured the microorganisms based on morphological characteristics. Three major plant pathogens (Fusarium oxysporum, Rhizoctonia solani, Phytophthora capsici) have been used to select antagonistic microorganisms. Total 20 bacteria and 9 fungi showed the pathogen growth suppression ability in vitro condition. The selected microorganisms were identified by ITS sequence similarity. CONCLUSION: All tested eco-friendly formulations contained high-density of the microorganisms. Among the isolated microorganisms, Bacillus spp. and Streptomyces spp. showed the most effective antifungal activity against the plant pathogens such as F. oxysporum, R. solani, and P. capsici. Among the selected fungi Trichoderma sp. demonstrated antifungal activity. Our results suggest that the currently adapted eco-friendly formulations might useful for sustain agricultural system.

• The Plant Pathology Journal
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• 제25권1호
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• pp.70-76
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• 2009

The rhizobacteria of Panax notoginseng were isolated from six sites in Yanshan, Maguan and Wenshan Counties, Yunnan Province of China, and their antagonistic activity against P. notoginseng root-rot fungal pathogens was determined. Of the 574 rhizobacteria isolated, 5.8% isolates were antagonistic in vitro to at least one of the five pathogens, Cylindrocarpon didynum, Fusarium solani, Phytophthora cactorum, Phoma herbarum, and Rhizoctonia solani. The number of rhizo bacteria and the number that inhibited fungi differed depending on sampling sites and isolation methods. Rhizobacteria isolated from the site in Yanshan and Maguan showed more antagonistic effect than them in Wenshan. Heat treatment of rhizosphere soil at $80^{\circ}C$ for 20 min scaled the antagonists up to 14.0%. Antagonistic bacteria in the roots proportioned 3.9% of the total isolates. The most antagonistic isolates 79-9 and 81-4 are Bacillus subtilis based on their 168 rDNA sequence and biochemical and physiological characteristics. Identification and evaluation of antagonistic bacteria against P. notoginseng root-rot pathogens in the main planting areas improved our understanding of their distribution in rhizosphere soil. Furthermore these results indicated that the interactions between biocontrol agent and soil microbes should be seriously considered for the successful survival and biocontrol efficacy of the agents in soil.

• 한국미생물·생명공학회지
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• 제28권6호
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• pp.334-340
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• 2000

In order to select multifunctional powerful antagonistic biocontrol agent against red-pepper rotting fungi Phytophthora capsici, we isolated an indigenous antagonistic bacterium which produces antifungal substances and siderophores from a local soil of Kyongju, Korea. The isolated strain was identified as Pseudomonas fluorescens biotype F. The antibiotic produced from P. fluorescens 2112 inhibited hyphae growth and the zoospore germination of Phytophthora capsici. The favorable carbon, nitrogen source and salts for the production of antibiotic from P. fluorescens 2112 were glycerol, beef extract and LiCi at 1.0%, 0.5% and 5 mM, respectively. And antagonistic activity of P. fluorescens 2112 was confirmed against P. capsici in vivo.

• Journal of Microbiology and Biotechnology
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• 제5권6호
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• pp.346-352
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• 1995

For isolation of antibiotic-producing bacteria antagonistic to Fusarium oxysporum, a total of 327 microorganisms were screened from sesame-growing soils collected at various locations in Korea by the modified Herr's triple-agar-Iayer technique. Among the 36 bacterial isolates further screened by the dual culture test on tryptic soy agar, 10 were tested to show their antagonistic activity against 14 plant pathogenic fungi. Bacterial culture filtrates were shown either to inhibit some phytopathogenic fungal growth or to suppress F. oxysporum infection of sesame plants maintained in the green house. An isolate, B23, with the most prominent antagonistic activity was identified as Bacillus subtilis.

• 한국환경농학회지
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• 제29권1호
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• pp.86-91
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• 2010

This study was carried out to assess the antifungal potential of R. oligosporus and its ethyl acetate (EtOAc) extract against the fungal pathogens causing anthracnose disease in apple fruits using disc diffusion, antagonistic effect and morphological abnormalities in fungal mycelia. The percentage of inhibition of antifungal effect of the ethyl acetate extract (5 ${\mu}l$ $disc^{-1}$) of the R. oligosporus against C. acutatum KACC 40848, C. gloeosporioides KACC 40897, C. higginsianum KACC 40806, C. orbiculare KACC 40808, C. coccodes KACC 40008, C. musae KACC 40947, C. boninense KACC 40893, C. liliacearum KACC 40981, C. caudatum KACC 41028 and Colletotrichum sp. KACC 40811 was found to be 44.4, 35.5, 40, 31.1, 33.3, 37.7, 40, 51.1, 28.8 and 28.8%, respectively. Also the fungus R. oligosporus showed potential antagonistic effect of antifungal activity against the tested pathogens of Colletotrichum spp. Further, R. oligosporus had a potential detrimental effect on the morphology of the tested fungi of Colletotrichum spp. such as wrinkle abnormalities, abnormal cell formation, lysis of mycelium, empty cell formation, distorted cell formation and breakage of the mycelium. These findings strongly support the role of R. oligosporus to serve as a potential antifungal agent to control plant pathogenic fungi causing anthracnose disease in apple fruits.

• 한국식물병리학회지
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• 제14권3호
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• pp.268-277
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• 1998

The production of Korean ginseng, one of the most important medicinal root crops, is limited by many factors including soil sickness, root rots in fields as well as during storage prior to consumption. Although much research has been conducted on the diseases in field condition, little information is available on the control of postharvest roots rots. To obtain better management strategy of postharvest root rots in ginseng, biological control using antagonistic bacteria was attempted. Of 208 bacteria obtained form suppressive soil samples, 4 were selected based on the inhibitory effect on mycelial growth of two major causal fungi for postharvest root rots in ginseng, Botrytis cinerea and Fusarium solani. The culture filtrates of these bacterial antagonists greatly inhibited the conidial germination of both pathogenic fungi and produced abnormal morphology such as swollen germ tubes in F. solani and vacuolation of nongerminated conidia in B. cinerea. The population levels of bacterial antagonists on the ginseng roots were gradually increased up to 8 days of incubation. Postharvest root rots of ginseng caused by f. solani and B. cinerea were controlled in dipping tests in the ranges of 60∼80% by antagonistic Bacillus spp. obtained from suppressive soil. These results suggest that biological control using these antagonistic bacteria would be an alternative strategy to control postharvest root rots in ginseng.

• The Plant Pathology Journal
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• 제15권5호
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• pp.280-286
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• 1999

Lichen-forming (LFF) or lichenicolous fungi (LCF) were isolated from the lichens collected at‘Backwoon’mountain area,‘Chiri’mountain area and‘Sorok’island in the southern regions of Korea and were screened for antagonistic efficacy against several phyto-pathogenic fungi. Symbiotic algae-free LFF and LCF were isolated by the following methods: I) discharged spores (ascospores), II) macerated thallus suspension and III) direct use of thallus fragments. Among 58 isolates obtained from 34 lichens, 8 isolates showed antifungal activity against Rhizoctonia solani. Antifungal activities of the strongest antagonistic isolate (LB9810) originated from the thallus of Parmelia quercina lichen were evaluated against 15 phyto-pathogenic fungi. When crude methanol extract of mycelia of the LB8910 isolate was employed at the rate of 0.5% (v/w), fungal growth of Magnaporthe grisea and Rhizoctonia solani was severly and Rhizoctonia solani was severly inhibited as much as approximately 60% compared to control. Growth of various food-borne same extract. The extract was successively partitioned with n-hexane, ethyl acetate and n-butanol. n-Hexane fraction displayed the strongest antifungal activities against R. solani. The LB9810 isolate was finally identified as Fusarium equiseti (Corda) Sacc., which has not been reported as LFF or LCF yet. Therefore, it is very likely that F. equiseti isolated it the study was originated from the contaminants associated with thallus fragments rather than from LFF or LCF.

• Mycobiology
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• 제40권1호
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• pp.59-65
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• 2012

Antagonistic microorganisms against $Rhizoctonia$ $solani$ were isolated and their antifungal activities were investigated. Two hundred sixteen bacterial isolates were isolated from various soil samples and 19 isolates were found to antagonize the selected plant pathogenic fungi with varying degrees. Among them, isolate C9 was selected as an antagonistic microorganism with potential for use in further studies. Treatment with the selected isolate C9 resulted in significantly reduced incidence of stem-segment colonization by $R.$ $solani$ AG2-2(IV) in Zoysia grass and enhanced growth of grass. Through its biochemical, physiological, and 16S rDNA characteristics, the selected bacterium was identified as $Bacillus$ $subtilis$ subsp. $subtilis$. Mannitol (1%) and soytone (1%) were found to be the best carbon and nitrogen sources, respectively, for use in antibiotic production. An antibiotic compound, designated as DG4, was separated and purified from ethyl acetate extract of the culture broth of isolate C9. On the basis of spectral data, including proton nuclear magneric resonance ($^1H$ NMR), carbon nuclear magneric resonance ($^{13}C$ NMR), and mass analyses, its chemical structure was established as a stereoisomer of acetylbutanediol. Application of the ethyl acetate extract of isolate C9 to several plant pathogens resulted in dose-dependent inhibition. Treatment with the purified compound (an isomer of acetylbuanediol) resulted in significantly inhibited growth of tested pathogens. The cell free culture supernatant of isolate C9 showed a chitinase effect on chitin medium. Results from the present study demonstrated the significant potential of the purified compound from isolate C9 for use as a biocontrol agent as well as a plant growth promoter with the ability to trigger induced systemic resistance of plants.