• 한국미생물·생명공학회지
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• 제23권1호
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• pp.98-103
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• 1995

During the screening of antifungal antibiotics from microbial metabolites, we selected Pseudomonas aeruginosa KGM-100 showing powerful antagonistic activity against various phytopathogenic fungi. Antibiotics KGM-100A and KGM-100B were purified from the culture broth of Pseudomonas aeruginosa KGM-100 by diaion HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, preparative TLC and recrystallization. KGM-100A which was recrystallized in MeOH showed antimicrobial activities against a broad spectrum of fungi and bacteria. Physico-chemical properties of KGM-100A were determined and identified to be phenazine-l-carboxylic acid by UV, IR, $^{1}$H-NMR, $^{13}$C-NMR, mass spectrum, and elemental analyses.

• 미생물학회지
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• 제13권1호
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• pp.1-23
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• 1975

This experiment was designed to elucidate the environmental factors in rhizosphers of bamboo forest that affect the distribution and the population size of Azotobacter, and also to estimate the annual productivities of nitrogen fixed by Azotobacter species. The results of this experiment can be summarized as follows ; The rhizosphere of bamboo forest contained high free sugars as of 3-8 times more than non-rhizosphere (Bacon, 1968), and the contents of organic matter and amino acids of that are reltively higher than this. Because of high content of potassium, average of soil pH is near at 7.0. As above-mentioned enviromental factors, the population sizes of Actinomycetes, general fungi, general bacteria and Azotobacters are larger than those of non-rhizosphere and the ofllowings are general fungi and general bacteria by turns. Azotobacter is dependent upon the antagonistic Actinomycetes. The main carbon source for Azotobacter in nitrogn flxation at the rhizosphere was glucose and minors were fructose, maltose and sucrose by turns. Annual gains of nitrogen by Azotobacters in soil of bamboo forest within 10cm from surface are estimated as of 88.94 kg/ha at site A, 60.4kg/ha at site B and 67.38kg/ha at site C, respectively.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2016년도 춘계학술대회 및 임시총회
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• pp.13-13
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• 2016

Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.

• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1361-1368
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• 2007

Endophytic bacteria associated with the roots of coastal sand dune plants were isolated, taxonomically characterized, and tested for their plant growth-promoting activities. Ninety-one endophytic bacterial isolates were collected and assigned to 17 different genera of 6 major bacterial phyla based on partial 16S rDNA sequence analyses. Gammaproteobacteria represented the majority of the isolates (65.9%), and members of Pseudomonas constituted 49.5% of the total isolates. When testing for antagonism towards plant pathogenic fungi, 25 strains were antagonistic towards Rhizoctonia solani, 57 strains were antagonistic towards Pythium ultimum, 53 strains were antagonistic towards Fusarium oxysporum, and 41 strains were antagonistic towards Botrytis cinerea. Seven strains were shown to produce indole acetic acid (IAA), 33 to produce siderophores, 23 to produce protease, 37 to produce pectinase, and 38 to produce chitinase. The broadest spectra of activities were observed among the Pseudomonas strains, indicating outstanding plant growth-promoting potential. The isolates from C. kobomugi and M. sibirica also exhibited good plant growth-promoting potential. The correlations among individual plant growth-promoting activities were examined using phi coefficients, and the resulting data indicated that the production of protease, pectinase, chitinase, and siderophores was highly related.

• The Plant Pathology Journal
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• 제22권1호
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• pp.103-106
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• 2006

In this biocontrol research, we evaluated disease suppressive effects of antagonistic bacterial strains ISE13 and KJ1R5 against Korean ginseng root rot caused by P. eaetorum. We also examined the effects of nutrient solution in the hydroponic culture system for Korean ginseng on biological activity of the bacterial strains. As results of dual culture tests of the bacterial strains on $V_{8}$ juice agar, the strain ISE13 showed antifungal activity against P. eaetorum and other plant pathogenic fungi, but the strain KJ1R5 did not. When their inhibitory effects against infection of P. eaetorum on the roots grown in either nutrient solution or water were tested, the strains ISE13 and KJ1R5 inhibited the disease severity of Korean ginseng roots only grown with water, compared to buffer-treated, inoculated controls. However, the nutrient solution used for hydroponic cultures of ginseng in pots caused higher levels of disease severity by the strains ISE13 and KJ1R5 from 418.8\%$ to 40.0\%$ and from 24.3\%$ to 45.0\%$, respectively. In this study, the bacterial strains ISE13 and KJ1R5 could be potentially biocontrol agents to suppress Korean ginseng root rot caused by P. eaetorum. However, more attention using nutrient solution in hydroponic cultures for Korean ginseng production should be applied in biocontrol of plant diseases using the antagonistic microorganisms.

• 한국환경농학회지
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• 제28권3호
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• pp.289-294
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• 2009

사과 재배에 피해를 주고 있는 사과 겹무늬썩음병, 탄저병, 라이족토니아병을 복합적으로 방제하기 위한 기초연구로서 각 병원균에 대하여 우수한 길항력을 동시에 가지는 길항미생물을 선발하고 Streptomyces sp. S-1110으로 동정하였다. 분리된 길항미생물은 배양상등액으로 길항물질을 분비하였으며 우수한 열안정성과 pH 안정성을 가지는 특성이 있었으며 각 병원균에 대한 감수성이 조금씩 달라서 2 종 이상의 길항물질이 동시에 생산되어 길항작용을 하는 것으로 추정되었다. 또한 분리균이 생산하는 길항물질을 사과 과실에 직접접종된 사과 겹무늬썩음병과 탄저병의 방제를 위해 실험한 결과 높은 방제력을 보였다. 따라서 분리된 길항미생물은 사과 과수원 및 저장소에서 동시에 수종의 질병방제에 쓰일 수 있는 천연 복합방제제의 개발을 위한 기초 균주로의 가치가 높은 것으로 판단된다.

• 한국식물병리학회지
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• 제14권3호
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• pp.191-202
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• 1998

About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

• Journal of Ginseng Research
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• 제40권2호
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• pp.97-104
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• 2016

Background: Rhizobacteria play an important role in plant defense and could be promising sources of biocontrol agents. This study aimed to screen antagonistic bacteria and develop a biocontrol system for root rot complex of Panax notoginseng. Methods: Pure-culture methods were used to isolate bacteria from the rhizosphere soil of notoginseng plants. The identification of isolates was based on the analysis of 16S ribosomal RNA (rRNA) sequences. Results: A total of 279 bacteria were obtained from rhizosphere soils of healthy and root-rot notoginseng plants, and uncultivated soil. Among all the isolates, 88 showed antagonistic activity to at least one of three phytopathogenic fungi, Fusarium oxysporum, Fusarium solani, and Phoma herbarum mainly causing root rot disease of P. notoginseng. Based on the 16S rRNA sequencing, the antagonistic bacteria were characterized into four clusters, Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetesi. The genus Bacillus was the most frequently isolated, and Bacillus siamensis (Hs02), Bacillus atrophaeus (Hs09) showed strong antagonistic activity to the three pathogens. The distribution pattern differed in soil types, genera Achromobacter, Acidovorax, Brevibacterium, Brevundimonas, Flavimonas, and Streptomyces were only found in rhizosphere of healthy plants, while Delftia, Leclercia, Brevibacillus, Microbacterium, Pantoea, Rhizobium, and Stenotrophomonas only exist in soil of diseased plant, and Acinetobacter only exist in uncultivated soil. Conclusion: The results suggest that diverse bacteria exist in the P. notoginseng rhizosphere soil, with differences in community in the same field, and antagonistic isolates may be good potential biological control agent for the notoginseng root-rot diseases caused by F. oxysporum, Fusarium solani, and Panax herbarum.

• The Plant Pathology Journal
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• 제31권3호
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• pp.278-289
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• 2015

Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, ${\beta}$-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

• 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.