Selection and Antifungal Activity of Antagonistic Bacterium Bacillus subtilis KMU-13 against Cucumber scab, Cladosporium cucumerinum KACC 40576

검은별무늬병균 Cladosporium cucumerinum KACC 40576에 대한 길항균주 Bacillus subtilis KMU-13의 선발 및 항진균 활성

  • 박성민 (계명대학교 미생물학과) ;
  • 이준석 (KBC생명공학연구소) ;
  • 박치덕 (대구 신기술사업단 전통생물소재산업화센터) ;
  • 이정훈 (계명대학교 미생물학과) ;
  • 정혁준 (계명대학교 미생물학과) ;
  • 유대식 (계명대학교 미생물학과)
  • Published : 2006.02.01

Abstract

Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

오이검은별무늬병균인 Cladosporium cucumerinum KACC 40576에 대하여 높은 항진균 활성을 나타내는 KMU-13은 Norway Lillehammer 근교 토양시료에서 분리하였으며, 형태학적, 배양 및 생화학적인 특성을 조사한 결과와 16S rDNA 염기서열을 분석한 결과, B. subtilis로 판단되었다. B. subtilis KMU-13의 항진균 물질의 생산을 위한 최적조건은 LB broth를 기본배지로 하여 탄소원으로 0.5% maltose와 질소원으로 0.5% bactopeptone을 첨가하였을 때 가장 높은 항진균 활성을 나타내었으며, 배양조건은 $30^{\circ}C$, 180rpm, 48시간이였고, 배양초기 pH는 6.0으로 조사되었다. TLC에서 Rf값이 0.64로 확인된 밴드에서만 항진균 활성을 가지는 것으로 조사되었으며 잿빛곰팡이병, 고추 탄저병, 참외 탄저병, 수박 등의 박과작물의 덩굴마름병, 토마토 시들음병, 글라디올러스 마른썩음병, 토마토 질병, 수박 덩굴쪼김병, 수박 질병, 사과나무 역병, 벼 잎집무늬마름병, 참외 줄기썩음병, 그리고 고추 균핵병에 대하여 양호한 항진균 활성을 나타내는 것으로 조사되었다.

Keywords

References

  1. Davison, J. (1998), Plant beneficial bacteria, Biotechnology 6, 282-286 https://doi.org/10.1038/nbt0388-282
  2. Lee, E. J., K. S. Kim, S. H. Hong, and J. H. Ha (1995), The mechanism of biological control of Pseudomonas spp. against Fusarium solani causing plant root-rot disease, Kor. J. Appl. Microbiol. Biotechnol. 23, 91-97
  3. Park, S. M., S. H. Han, and T. S. Yu (2005), Culture conditions and antifungal activity of Bacillus licheniformis KMU-3 against crop pathogenic fungi, Kor. J. Appl. Microbial. Biotechnol. 33, 112-116
  4. Park, S. M., H. J. Jung, S. H. Han, S. H. Yeo, Y. W. Kim, H. G. Ahn, H. S. Kim, and T. S. Yu (2005), Antifungal activity of extract from Xanthium strumarium L. against plant pathogenous fungi, JLS. 15, 692-695 https://doi.org/10.5352/JLS.2005.15.5.692
  5. Oh, Y. J. (1992), Studies on the optimization of media composition and cultural conditions for kasugamycin production by Streptomyces kasugansis, Kor. J. Appl. Microbial. Biotechnol. 20, 583-587
  6. Bae, M. (1978), Present status and future of antibiotics for agriculture, Kor. J. Appl. Microbial. Biotechnol. 6, 141-148
  7. Reveni, M., H. Cohen, T. Zahavi, and V. Venezion (2000), Polar a potant polyxin B compound for controlling powdery mild in apple and nectarine trees, and grapevins, Crop protection. 19, 393-399 https://doi.org/10.1016/S0261-2194(00)00030-2
  8. Edward, J. K., R. P. Adams, and K. K. Kartha (1990), Trehalase activity in plant tissue culture, Phytochemistry 29, 2525-2528 https://doi.org/10.1016/0031-9422(90)85180-N
  9. Ko, B. S., T. Oritani, and K. Yamashita (1992), Synthesis and biological activity of 6-phenylgriseofulvin as analogs of antibiotic griseofulvin, J. Kor. Agric. Chem. Soc. 35, 395-398
  10. Kim, S. U., J. W. Lee, S. H. Lee, and S. H. Bok (1991), Identification of bacteria having antifungal activity isolated from solid and its activity, Kor. J. Microbiol. Biotechnol. 19, 337-342
  11. Shin, Y. J., M. J. Jung, J. U. Park, W. H. Joo, and Y. K. Jeong (2000), Isolation and identification a bacterium producing antifungal antibiotic, Kor. J. Food. Sci. Nutr. 29, 832-836
  12. Holt, J. G., N. R. Krieg, P. H. A. Sneath, J. T. Staley, and S. T. Williams. (1994), Bergey's Manual of Systematic Bacteriology. 9th. Williams&Wilkins. USA
  13. Thompson, J. D., D. G. Higgins, and T. J. Gibson (1994), CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalies and weight matrix choice, Nucleic Acids Res. 22, 4673-4680 https://doi.org/10.1093/nar/22.22.4673