• 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.

• 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.

• 한국잔디학회:학술대회논문집
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• 한국잔디학회 2011년도 제24차 정기총회 및 학술발표회
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• pp.35-35
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• 2011

A large patch disease caused by Rhizoctonia solani AG2-2(IV) is a serious problem in turfgrass sites including golf courses and sports fields in Korea. The objectives of this study were to isolate some antagonistic microorganisms and to explain some involving mechanisms. Initially single colonies which were formed from the filtrates of various soil samples were obtained from LB culture and then co-cultured with R.solani AG2-2(IV) on PDA plate to explore some antagonistic microbes against for large patch fungus, Rhizoctonia solani AG2-2(IV). Out of total 82 antagonistic isolates which commonly had inhibition effect on Rhizoctonia solani AG2-2(IV) mycelial growth, one candidate (YPIN22) showed the most antifungal effect, which was confirmed by the longest distance from the edge of bacterial colony to the mycelial edge of the Rhizoctonia solani AG2-2(IV) in the dual culture. A succeeding investigation was to test any potential effect of the isolate on growth inhibition of 5 other turfgrass pathogens including R. solani solani AG2-2(IIIB), P. ultimum, C. caudatum, C. lunata, and F.oxysporum. Preliminary result indicated that the new isolate YPIN22 was also found to have antagonistic potential on the growth inhibition of those turfgrass pathogenic fungi, which was explained by inhibition zones ranging from 8 to 22mm. A further explanation of some characteristics of the isolate YPIN22 will be discussed in detail.

• Journal of Applied Biological Chemistry
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• 제42권4호
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• pp.197-201
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• 1999

Antagonistic bacteria active against phytopathogenic fungi, Phytophthora capsici, Pythium ultimum, Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum were isolated from greenhouse soils. An antifungal compound was extracted by ethyl acetate from acidified culture filtrate and purified through column chromatography and thin layer chromatography. Activity-guided bioassay was followed throughout the purification steps using Pythium ultimum as a test organism. The purified antifungal compound was identified as phenylacetic acid (PAA) based on the data obtained from IR, EI/MS, $^1H-NMR$, and $^{13}C-NMR$. Two different isolates, which had vast differences in differential characteristics except 16S rDNA sequence homology, produced the same compound, phenylacetic acid. $ED_{50}$ values of the phenylacetic acid against P. ultimum, P. capsici, R. solani, B. cinerea, and F. oxysporum were 45, 21, 318, 360, and 226 ppm, respectively.

• The Plant Pathology Journal
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• 제24권1호
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• pp.52-57
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• 2008

Twenty isolates of Bacillus spp. obtained from livestock manure composts and cotton-waste composts were tested for in vitro antagonistic effects against soilborne plant pathogenic fungi, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani AG-4, and Sclerotinia sclerotiorum. Seven isolates of Bacillus spp. had antagonistic effects on mycelial growth of all the isolates of F. oxysporum tested. The bacterial isolate RM43 was the most effective to inhibit the mycelial growth of the fungal isolates. Twelve isolates of Bacillus spp. had antagonistic effects on mycelial growth of all the isolates of P. capsici tested. The bacterial isolates M34 and M47 were very effective to inhibit the mycelial growth of the fungal isolates. Thirteen isolates of Bacillus spp. had antagonistic effects on mycelial growth of all the isolates of R. solani AG-4 tested. The bacterial isolates M27 and M75 were very effective to inhibit the mycelial growth of the fungal isolates. Fourteen isolates of Bacillus sp. had antagonistic effects on mycelial growth of all the isolates of S. sclerotiorum tested. The bacterial isolates M49 and M75 were very effective to inhibit the mycelial growth of the fungal isolates. The antagonistic effects of most Bacillus spp. isolates against the isolates of the four fungi differed depending on the fungal species and the isolates of each fungus. The bacterial isolates M27 and M75 were the most effective to inhibit the mycelial growth of all four fungi.

• 한국미생물·생명공학회지
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• 제29권3호
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• pp.142-148
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• 2001

경주 인근 지역의 토양으로부터 식물병원성 진균 Fusarium oxysprum과 phytophthora capsici를 동시에 길항할수있는 항진균성 항생물질 생산성 생물방제균을 분리하고, 성분을 함유한 병원진균의 세포벽을 분해하는 chitinase 생산성이 우수한 균주를 분리하고자 하였다. 분리된 생물방제균의 형태학적, 생화학적 및 배양학적으로 동정하여 잠정적으로 Bacillus amyloliquefaciens 7079로 동정하였다. 이 생물방제균이 생산하는 chitinase 생산의 최적 조건을 검토하여 본 결과 Chitin-yeast extract 배지(0.7% $K_2$$HPO_4$ 0.2% $KH_2$$PO_4$ 0.1%($NH_4$)$_2$$SO_4$ 0.05% sodium citrate 0.01% MgSO$_4$$7H_2$O 0.1% yeast extract, 0.1% coloida chitin)에서 pH는 7.0 배양온도는 3$0^{\circ}C$였고 배양한 후 3일 이 되었을 때 가장 많은 chitinase를 생산하였다. 또한 0.1% colloida chitin을 탄소원으로 하여 배양하였을 때 chitinase 생산성이 가장 좋았으며 0.15% proteose peptone NO .3 또는 0.1% tryptone 을 질소원으로 하여 배양하였을 때 효소 생산이 높게 나타났다. 선발된 생물방제균의 고추를 기주식물로한 in vivo pot 시험 결과 고추역병균 Phytophthora capsici에 좋은 길항력을 확인할 수 있었다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.96.2-97
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• 2003

More than 1200 microorganisms were isolated from soil samples collected from various sources and localities. Among the isolates, 2 actinomyces (TH-04 and BA313) and 1 Bacillus sp. (CJ3) were selected as antagonists to pepper anthracnose fungus Colletotrichum gloeosporioides. These 3 isolates inhibitied mycelial growth of C gloeosporioides and the inhibition rates were over 70％ on PDA. When the isolates were co-cultured with conidia of C. gloeosporioides in potato dextrose broth, conidial germination was severely inhibited and the inhibition rates of TH-04, BA313, and CJ3 at 24 hours were 75％, 72％, and 68％, respectively. The inhibition rates at n hours incubation were not much different from the rates at 24 hours. To check the activity on the plant, each isolate was mixed with equal volume of conidial suspension of C. gloeosporioides and wound-inoculated on green pepper fruit. After 6 days, the anthracnose lesions on the fruits inoculated with the mixture were much smaller than the lesions caused by the C. gloeosporioides itself. The lesion areas of TH-04 or BA313 treated pepper were less than 30％ of the check. TH-04 and BA313 also showed antagonistic activity to Phytophthora spp. and Botrytis cinerea. By scanning electron microscopy and fatty acid analyses (MIDI), TH-04 and BA313 were identified to Streptomyces halstedii and S. violaceusniger, repectively.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.984-988
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• 2003

The actinomycete strain KH-6l4 possessed strong antifungal activity, especially antagonistic to the rice blast fungus, Pyricularia oryzae. Diaminopimelic acid (DAP) type and morphological and physiological characteristics, examined by scanning electron microscopy (SEM), indicated that KH-614 belonged to the genus Streptomyces. Antifungal agent produced by this strain was found to be most active, when the strain was cultured in the presence of glucose, polypeptone, and yeast extract (PY) medium for 6 days at $27^{\circ}C$. Based on the spectral report data, MS and NMR, the antifungal agent was identified as cyclo(L-leucyl-L-prolyl). According to the antimicrobial activity test measured by minimal inhibitory concentration (MIC), the cyclo(1eu-pro) exhibited the activity against Candida albicans IAM 4905, Mucor ramannianus IAM6218, Rhizoctonia solani IFO 6218, Aspergilus fumigatus ATCC 42202, Glomerella cingulata IFO 9767, Trichophton mentagrophytes ATCC 18749, and Trichophyton rubrum ATCC 44766, the order of MIC values were 50, 12.5, 5, 50, 25, 5, $5\;\mu\textrm{g}/ml$, respectively. Specifically, cyclo(1eu-pro) was one of the most effective elements against Pyricularia oryzae IFO 5994 with the MIC value of $2.5\;\mu\textrm{g}/ml$, thus indicating that cyclo(leu-pro) is a potential antifungal agent.

• Mycobiology
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• 제49권5호
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• pp.498-506
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• 2021

An endophytic fungus strain DYSJ3 was isolated from a stem of Aphanamixis grandifolia Blume, which was identified as Aspergillus versicolor based on the morphological characteristics, internal transcribed spacer (ITS) and calmodulin gene sequences analyses. A. versicolor DYSJ3 exhibited strong antagonistic activity against Colletotrichum musae, C. gloeosporioides and Fusarium oxysporum f. sp. cubense with the inhibition rates of 61.9, 51.2 and 55.3% respectively. The antifungal metabolites mainly existed in the mycelium of A. versicolor DYSJ3, and its mycelial crude extract (CE) had broad-spectrum antifungal activities against plant pathogenic fungi. The CE had a good thermal stability, and the inhibition rate of 100 mg/mL CE against C. musae was above 70.0% after disposing at 120 ℃ for 1 h. Five secondary metabolites were isolated from the CE and identified as averufanin, ergosterol peroxide, versicolorin B, averythrin and sterigmatocystin. Activity evaluation showed versicolorin B exhibited inhibitory effects on the mycelial growth and conidial germination of C. musae, and sterigmatocystin had a weak inhibitory effect on the mycelial growth of C. musae.

• Journal of Microbiology and Biotechnology
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• 제13권3호
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• pp.415-421
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• 2003

A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.