Bacillus subtilis HJ927에 의해 생성된 화합물의 분리, 동정 및 고추(Capsicum annum L.) 역병방제 효과

Isolation and Identification of Low Molecular Weight Compounds Produced by Bacillus subtilis HJ927 and Their Biocontrol Effect on the Late Blight of Pepper (Capsicum annuum L.)

  • 이현진 (전남대학교 생물환경과학부) ;
  • 박근형 (전남대학교 생물환경과학부) ;
  • 심재한 (전남대학교 생물환경과학부) ;
  • 박노동 (전남대학교 생물환경과학부) ;
  • 김용웅 (전남대학교 생물환경과학부) ;
  • 황보훈 (전남대학교 응용생물공학부) ;
  • 조정용 (전남대학교 생물환경과학부) ;
  • 김영철 (전남대학교 응용식물학부) ;
  • 김길용 (전남대학교 생물환경과학부)
  • Lee, Hyun-Jin (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Park, Keun-Hyung (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Shim, Jae-Han (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Park, Ro-Dong (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Kim, Yong-Woong (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Hwang-Bo, Hoon (Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, APSRC, Chonnam National University) ;
  • Cho, Jeung-Yong (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University) ;
  • Kim, Young-Cheol (Division of Applied Plant Science, College of Agriculture and Life Science, Chonnam National University) ;
  • Kim, Kil-Yong (Glucosamin Saccharide Material Laboratory(NRL), Division of Applied Bioscience and Biotechnology, Chonnam National University)
  • 투고 : 2004.11.08
  • 심사 : 2004.12.14
  • 발행 : 2005.02.28

초록

병원성 곰팡이에 대해 강한 길항성을 보이는 토양미생물 Bacillus subtilis HJ927을 Phytophthora capsici에 감염된 고추밭으로부터 분리해냈다. B. subtilis HJ927을 P. capsici와 함께 고추에 처리한 결과 P. capsici만을 접종한 처리구에 비해 크게 식물을 보호하는 것을 root mortality 측정결과 확인하였다. B. subtilis HJ927는 항곰팡이성 물질로 3-methylbutyric acid, 2-methylbutyric acid, 그리고 methyl 2-hydroxy, 3-phenyipropanoate를 분비해 내는 것을 HPLE와 GC-MS를 통해 분리 동정하였다. 또한 B. subtilis HJ927는 가수분해효소인 ${\beta}$-1,3-glucanase를 분비해 냄으로서 위 화합물과 함께 식물을 보호하는 것으로 분석되었다.

A soil bacterium, Bacillus subtilis HJ927, exhibiting strong antagonistic property against pathogenic fungi was isolated from pepper fields infested with Phytophthora capsici. Pepper plants inoculated with P. capsici revealed severe root mortality while plants co-inoculated with B. subtilis HJ927 and P. capsici showed drastically reduced root mortality. Low molecular weight substances released by B. subtilis HJ927 mediated the plant protective effect. The anti-fungal compounds released by B. subtilis HJ927 were identified as 3-methylbutyric acid, 2-methylbutyric acid, and methyl 2-hydroxy, 3-phenylpropanoate by high-performance liquid chromatography and gas chromatography-mass spectrometry. In addition to these compounds, B. subtilis HJ927 also produced ${\beta}$-1,3-glucanase, a hydrolytic enzyme implicated in antifungal activity.

키워드

참고문헌

  1. Asante, G. S., and A. L. Neal. 1964. Characterization of fungistatic substances produced by a Bacillus antagonistic to Ceratocystis ulmi. Phytophathol. 54:819-822
  2. Babad, J., A. Pinsky, R. Turner Graff, and N. Sharon. 1952. An antifungal polypeptide produced by Bacillus subtilis. Nature 170:618-619 https://doi.org/10.1038/170618a0
  3. Baker, C. J., J. R. Stavely, C. A. Thomas, M. Sasse, and J. S. MacFall. 1983. Inhibitory effect of Bacillus subtilis on Uromyces phaseoli and on development of rust pustules on bean leaves. Phytopathol. 73: 1148-1152 https://doi.org/10.1094/Phyto-73-1148
  4. Dumas-Gaudot, E., S. Siezack, B. Dassi, M. J. Pozo, V. Gianinazzi-Pearson, and S. Gianinazzi. 1996. Plant hydrolytic enzymes (chitinase and $\beta$-1,3-glucanase) in root reactions to pathogenic and symbiotic microorganisms. Plant Soil 185:211-221 https://doi.org/10.1007/BF02257526
  5. Fogliano, V., A. Ballio, M. Gallo, S. Woo, R. Scala, and M. Lorito. 2002. Pseudomonas lipodepsipeptides and fungal cell wall-degrading enzymes act syringistically in biological control. Mol. Plant-Microbe Interact. 15:323-333 https://doi.org/10.1094/MPMI.2002.15.4.323
  6. Fravel, D. R. 1998. Role of antibiosis in the biocontrol of plant diseases. Ann. Rev. Phytopathol. 26:75-91
  7. Hwang, B. K, and B. S. Kim. 1995. In-vivo efficacy and in-vitro activity of tubercidine, an antibiotic nucleoside, for control of Phytophthora capsici blight in Capsicium annuum. Pestic. Sci. 44:255-260 https://doi.org/10.1002/ps.2780440308
  8. Hwang, B. K, and D. H. Kim. 1995. Phytophthora blight of pepper and its control in Korea. Plant Dis. 79:221-227 https://doi.org/10.1094/PD-79-0221
  9. Hwang, B. K., S. W. Lim, B. S. Kim, J. Y. Lee, and S. S. Moon. 2001. Isolation and in vivo and in vitro antifungal activity of phenylacetic acid and sodium phenylactate from Streptomyces humidus. Appl. Environ. Microbiol. 67:3739-3745 https://doi.org/10.1128/AEM.67.8.3739-3745.2001
  10. Katz, E., and A. L. Deamain. 1977. The peptide antibiotic of Bacillus: Chemistry, biogenesis, and possible functions. Rev. Bacteriol. 41:449-474
  11. Kim, K. D., S. Nemec, and G. Musson. 1997. Control of phytophthora root and crown rot of bell pepper with composts and soil amendments in the greenhouse. Appl. Soil Bcol. 5:169-179 https://doi.org/10.1016/S0929-1393(96)00138-2
  12. Kim, K. Y., Y. S. Cho, B. K Sohn, R. D. Park, J. H. Shim, S. J. Jung, Y. W. Kim, and K Y. Seong. 2002. Cold-storage of mixed inoculum of Glomus intraradices enhances root colonization, phosphorus status and growth of hot pepper. Plant Soil 238:267-272 https://doi.org/10.1023/A:1014474617170
  13. Kobayashi, D. Y., M. Guglielmoni, and B. B. Clarke. 1995. Isolation of the chitinolytic bacteria xantomonas maltophilia and serratia marcescens as biological control agents for summer patch disease of turfgrass. Soil Biol. Biochem. 27:1479-1487 https://doi.org/10.1016/0038-0717(95)00062-J
  14. Larena, I., and P. Melgarejo. 1996. Biological control of Monilinia laxa and Fusarium oxysporeum f. sp. lycopersiciby a lytic enzyme-producing Penicillium purpurogenum. Biol. Control 6:361-367 https://doi.org/10.1006/bcon.1996.0046
  15. Liu, X., and B. Huang. 2000. Carbohydrate accumulation in relation to heat stress tolerance in two creeping bentgrass cultivars. J. Amer. Soc. Holt. Sci. 125:442-447
  16. Mavingui, P. and T. Heulin. 1994. In vitro chitinase and antifungal activity of a soil, rhizoplane population of Bacillus polymyxa. Soil Biol. Biochem. 26:801-803 https://doi.org/10.1016/0038-0717(94)90277-1
  17. Mckeen, C. D., C. C. Reillyd., and P. L. Pusey. 1986. Production and partial characterization of antifungal substances antagonistic to Monilinia fructicola from Bacillus subtilis. Phytopathol. 76: 136-139 https://doi.org/10.1094/Phyto-76-136
  18. Melent'ev, A. I., G. E. Akutganov., and N. F. Galimzyanova. 2001. The role of chitinase in the antifungal acitivity of Bacillus sp. 739. Microbiol. 70:548-552 https://doi.org/10.1023/A:1012304004659
  19. Michener, H. D., and N. Snell. 1949. Two antifungal substances from Bacillus subtilis cultures. Arch. Bioehem. 22: 208-214
  20. Poxo, M. J., C. Cordier, E. Dumas-Gaudot, S. Gianinazzi, J. M. Barea, and C. Azcon Aguilar. 2002. Localized versus systemic effect of arbuscular mycorrhizal fungi on defense responses to Phytophthora infection in tomato plants. J. Exp. Bot. 53:525-534 https://doi.org/10.1093/jexbot/53.368.525
  21. Ryder, M. H., Z. Yan, T. E. Terrace, A. E. Rovira, W. Tang, and R. L. Cerrell. 1999. Use of strains of Bacillus isolated in china to suppress take all and rhizoctonia root rot, and promote seedling growth of glasshouse-grown wheat in Australian soils. Soil Biol. Biochem. 31:19-29 https://doi.org/10.1016/S0038-0717(98)00095-9
  22. Singh, P. P., Y. C. Shin, C. S. Park, and Y. R. Chung. 1999. Biological control of Fusarium wilt of cucumber by chitinolytic bacteria. Phytopathol. 89:92-99 https://doi.org/10.1094/PHYTO.1999.89.1.92
  23. Tang, W. H. 1994. Yield increasing bacteria and biological control of sheath blight of rice. p. 267-273. In M. H. Ryder et al. (ed.) Improving plant productivity with rhizosphere bacteria. Adelaide, CSIRO, Australia
  24. Wei, G., J. W. Klopper, and S. Tuzun. 1991. Induction of systemic resistance of cucumber to Colletotrichum orbiculare by select strains of plant growth promoting rhizobacteria. Phytopathol. 81:1508-1512 https://doi.org/10.1094/Phyto-81-1508
  25. Weller, D. M., W. J. Howie, and R. J. Cook. 1998. Relationship between in vitro Gaeumannomyces graminis var. tritici and suppression of take all of wheat by fluorescent Pseudomonas. Phytophathol. 78:1094-1100