Selection and Mechanisms of Indigenous Antagonistic Microorganisms against Sheath Rot and Dry Rot Disease of Garlic

마늘 잎집썩음병과 마른썩음병을 길항하는 토착길항미생물의 선발 및 기작

  • Jeong, Hee-Young (Department of Applied Microbiology & Biotechnology, College of Natural Resources, Yeungnam University) ;
  • Lim, Jong-Hui (Department of Applied Microbiology & Biotechnology, College of Natural Resources, Yeungnam University) ;
  • Kim, Byung-Keuk (Department of Applied Microbiology & Biotechnology, College of Natural Resources, Yeungnam University) ;
  • Lee, Jung-Jong (Yeongcheonsi Agritechnology Center) ;
  • Kim, Sang-Dal (Department of Applied Microbiology & Biotechnology, College of Natural Resources, Yeungnam University)
  • 정희영 (영남대학교 응용미생물학과) ;
  • 임종희 (영남대학교 응용미생물학과) ;
  • 김병극 (영남대학교 응용미생물학과) ;
  • 이중종 (영천시농업기술센터) ;
  • 김상달 (영남대학교 응용미생물학과)
  • Received : 2010.05.19
  • Accepted : 2010.06.16
  • Published : 2010.09.28

Abstract

Sheath rot and dry rot disease caused by Pseudomonas marginalis and Fusarium oxysporum were serious problems in garlic farmland. In this study, total of 160 indigenous antagonistic bacteria were isolated from 16 farmlands in Yeongcheon, Korea. Among these, 15 strains were able to inhibited P. marginalis and F. oxysporum. The 16s rDNA genes of the selected 15 strains were amplified and sequenced. The strains has strong antagonistic ability against garlic pathogens was achieved Bacillus subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240. The selected 3 strains tested for investigation of antifungal mechanisms further analyses; 3 strains of these validated for production of siderophore, ${\beta}$-glucanase and chitinase using CAS (chrome azurol S) blue agar, CMC-congo red agar and DNS method. The 3 strains were able to utilized insoluble phosphate as dertermined by vanado-molybdate method. The 3 strains verified for production of auxin and gibberellic acid using Salkowski test and holdbrook test. Also, 3 strains showed stimulation germination, stem growth promoting activity on the in vivo test. The 3 strains were able to effectively suppress P. marginalis and F. oxysporum causing sheath rot and dry rot diseases on the in vivo pot test.

마늘병원균에 대한 토착길항미생물 선발을 위하여 경북 영천시 신덕리 및 구계리 일대 16개소의 토양 시료로부터 160균주의 세균들을 분리하였다. 이들 분리된 세균 중 마늘 잎집썩음병의 원인균인 P. marginalis와 마른썩음병의 원인균인 F. oxysporum에 대하여 길항능이 매우 뛰어난 3종의 토착길항세균을 선발하였다. 선발 균주들을 16s rDNA sequencing과 Bergey's manual을 이용한 방법에 의해 동정한 결과 B. subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240 균주들로 동정할 수 있었다. 선발된 3종의 토착길항세균들은 항세균 및 항진균 길항기작인 siderophore, ${\beta}$-glucanse, chitinase, 항생물질 생산능을 가진 균주들이었으며, 식물의 생장촉진에 도움이 되는 인산가용 화능과 식물생장촉진호르몬인 옥신 및 지베렐린을 생산하는 균주들로 확인되었다. 또한, 3종의 토착길항세균들은 in vivo 포트실험을 통해 잎집썩음병원균인 P. marginalis 에 대한 방제율이 70% 이상이고 동시에 생장촉진능도 있다는 것을 검증하였다. 그리고 마른썩음병원균인 F. oxysporum에 대한 방제율도 B. subtilis YC82와 B. amyloliquefaciens Y240의 경우 60%, B. vallismortis YC84는 80% 이상으로 나타났다. 또한, 발아촉진능에서는 선발된 토착길항세균의 처리구가 무처리구에 비하여 20% 이상 향상된 우수한 발아촉진능을 나타내었으며, 생장촉진능 역시 초장 길이의 차이에서 B. subtilis YC82는 평균 $49{\pm}1.53\;mm$, B. vallismortis YC84 는 평균 $47{\pm}1.15\;mm$, B. amyloliquefaciens YC240에서는 평균 $80{\pm}1.21\;mm$로 무처리구의 평균 $39{\pm}1.51\;mm$ 보다 매우 우수하였다. 이와 같은 결과는 선발된 마늘잎집썩음병 및 마른썩음병 방제용 토착길항세균은 마늘병원균 방제능과 동시에 생장촉진능을 가지는 다기능 토착길항세균으로 향후 추가적인 현장적용시험을 통하여 병방제능 및 생장촉진능 검증이 이루어 진다면 마늘병해방제 및 생장촉진용 미생물제제 개발이 가능할 것이라 생각된다.

Keywords

References

  1. Bhat, K. M. and R. Maheshwari. 1987. Sporotrichum thermophile growth, cellose degration and cellulase activity. Appl. Env. Microbio. 53: 2175-2182.
  2. Choi, J. E. and K. S. Han. 1990. Studies on the bacterial soft rot disease of lilliaceae crops 3. rot of garlic caused by Pseudomonas spp. Korean J. Plant Pathol. 6: 86-90.
  3. Claus D and R. C. W. Berkeley. 1986. Genus Bacillus. In Bergeys's Manual of Systemic Bacteriology. pp. 1105-1139. Williams & Wilkins, Baltimore, MD, USA.
  4. Gutierrez-Manero, F. J., B. Ramos-Solano, A. Probanza, J. Mehouachi, F. R. Tadeo, and M. Talon. 2001. The plantgrowth- promoting rhizobacteria Bacillus pumilus and Bacillus licheniformis produce high amounts of physiologically active gibberellins. Physiol Plant. 111: 206-211. https://doi.org/10.1034/j.1399-3054.2001.1110211.x
  5. Jeuniaux, C. 1966. Methods in enzymology 8, pp. 644-650. Academic Press, New York.
  6. Jung, H. K., J. R. Kim, S. M. Woo, and S. D. Kim. 2006. An auxin producing plant growth promoting rhizobacterium Bacillus subtilis AH18 which has siderophore-producing biocontrol activity. Kor. J. Microbiol. Biotechnol. 34: 94-100.
  7. Jung, H. K., S. C. Park, B. K. Park, S. D. Kim, D. H. Nam, and J. H. Hong. 2008. Characteristics of the antibacterial substance produced by Paenibacillus polymyxa JB115. Korean J. Biotechnol. Bioeng. 23: 65-69.
  8. Kim, D. H., H. K. Jung, and S. D. Kim. 2004. Selection and identification of auxin-producing plant growth promoting rhizobacteria having phytophathogen antagonistic activity. J. Kor. Soc. Appl. Biol. Chem. 47: 17-21.
  9. 김동환, 채성훈, 강명구. 2007. 마늘산업 발전 방안 연구. 농식품신유통연구원.
  10. Kim, Y. K., S. B. Lee, S. S. Lee, H. S. Shim, and I. H. Choi. 2003. Cultural and chemical approaches for controlling postharvest diseases of garlics. The Korean Journal of Pestcide Science. 7: 139-148. https://doi.org/10.1080/12265071.2003.9647696
  11. Lee, J. J., H. S. Lim, T. H. Chang, and S. D. Kim. 1999. Isolation of siderophore-producing Pseudomonas fluorescens GL7 and its biocontrol activity against root-rot disease. Kor. J. Appl. Microbiol. Biotech. 27: 427-432.
  12. Lee, J. T., D. H. Kim, J. H. Do, and S. D. Kim. 1998. Purification and characterization of chitinase from antagonistic bacteria Pseudomonas sp. 3098. Kor. J. Appl. Microbiol. Biotech. 26: 515-522.
  13. Lee, Y. S., H. J. Son, I. H. Kim, and T. I. Mheen. 1983. Studies on the production of gibberellic acid. Kor J Appl Microbiol Bioeng. 11: 217-222.
  14. Lim, J. H. 2009. Plant growth promotion, biological control, abiotic stress resistance induction and in situ monitoring by multi-functional plant growth promoting rhizobacteria. Graduate School, Yeungnam University.
  15. Lim, J. H. and S. D. Kim. 2009. Selection and characterization of the bacteriocin-producing bacterium, Bacillus subtilis BP6 isolated from chicken gut against Salmonella gallinarum causing fowl-typhus. J. Korean Soc. Appl. Biol. Chem. 52: 80-87. https://doi.org/10.3839/jksabc.2009.014
  16. Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-428. https://doi.org/10.1021/ac60147a030
  17. Oh, J. Y., Y. H. Lee, Y. D. Jin, J. B. Kim, S. G. Hwang, S. H. Han and J. E. Kim. 2007. Antifungal activity of pestcides to control dry rot and blue mold during garlic storage. The Korean Journal of Pestcide Science. 11: 331-338.
  18. Rural development administration and Korea forest service. 2008. Diagnosis and control of insect pests and crops diseases. Haksulpyeonsokwan.
  19. Shivanna, M. B., M. S. Meera, and M. Hyakumachi. 1994. Sterile fungi from zoysia grass rhizosphere as plant growth promoters in spring wheat. Can. J. Microbiol. 40: 637-644. https://doi.org/10.1139/m94-101
  20. Yang, M. J., S. H. Jung, E. S. Shin, J. H. Kim, H. D. Yun, S. L. Wong, and H. Kim. 2004. Expression of a Bacillus subtilis endoglucanase in protease-deficient Bacillus subtilis strains. J. Microbiol. Biotechnol. 14: 430-434.