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길항미생물 Bacillus subtilis GG95를 이용한 상추 균핵병의 생물학적 방제

Biological Control of Lettuce Sclerotinia Rot by Bacillus subtilis GG95

  • 이현주 (경기도농업기술원 환경농업연구과) ;
  • 김진영 (경기도농업기술원 소득자원연구소) ;
  • 이진구 (경기도농업기술원 소득자원연구소) ;
  • 홍순성 (경기도농업기술원 종자관리소)
  • Lee, Hyun-Ju (Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services) ;
  • Kim, Jin-Young (Agricultural resources Research Institute, Gyeonggido Agricultural Research & Extension Services) ;
  • Lee, Jin-Gu (Agricultural resources Research Institute, Gyeonggido Agricultural Research & Extension Services) ;
  • Hong, Soon-Sung (Variety Service Center, Gyeonggido Agricultural Research & Extension Services)
  • 투고 : 2014.07.31
  • 심사 : 2014.08.31
  • 발행 : 2014.09.30

초록

상추재배시 문제가 되고 있는 균핵병과 시들음병에 대한 미생물 방제제 개발을 위하여 병해가 발생된 상추 포장에서 미생물을 분리하고, 길항미생물을 선발하여 균의 특성을 조사하고 균핵병에 대한 예방효과를 검정한 결과는 다음과 같다. 경기도 주요 상추 재배지에서 균핵병은 10월부터 발생하여 다음해 5월까지 발생하였으며, 12월에는 최대 40%까지 발생하여 피해를 주고 있었다. 균핵병과 시들음병에 모두 억제효과가 있는 균주를 대치배양을 통해 선발하였다. 선발됨 균주는 Vitek 2의 BCL카드를 이용한 생화학적반응과 형태적 확인, 16s rRNA 분석을 통해 동정한 결과 Bacillus subtilis로 동정되었고 B. subtilis GG95로 명명하였다. B. subtilis GG95의 배양특성을 조사한 결과 온도 $25{\sim}30^{\circ}C$, 산도 7의 배양조건에서 배양이 잘 되었으며, 배양에 적정한 영양원은 탄소원 전분, 질소원 tryptone과 yeast extract를 무기염류 $MgCl_2$를 사용하였을 때 가장 잘 되었다. 영양원에 따라 균핵병과 시들음병에 대한 길항효과가 좋은 탄소원은 fructose, maltose였고 질소원은 peptone였으며, 무기염류는 균핵병은 $CaCl_2$, $K_2HPO_4$, 시들음병은 $MgSO_47H_2O$로 병원균에 대한 길항효과가 차이가 있었다. 상추 균핵병에 대한 B. subtilis GG95의 포장검정 결과 $1{\times}10^8cfu/mL$ 농도의 길항미생물 배양액을 포기당 10 mL씩 3회 관주하고 균핵병원균을 접종한 후 균핵병의 발병정도를 조사한 결과 88% 방제효과가 나타났다.

Sclerotinia sclerotiorum, a plant pathogenic fungus, can cause serious yield and quality losses in the winter lettuce field. For biological control of S. sclerotiorum, soil-born microorganisms that inhibit the mycelia growth of S. sclerotiorum and Fusarium oxysporum were isolated from diseased soil. Among the isolates, bacterial isolate, GG95, which was identified as Bacillus subtilis according to the morphological, physiological characteristics and by 16S rRNA similarity, showed the highest level of inhibitory activity. The growth conditions for B. subtilis GG95 were optimized in TSB media (pH 7) by culturing at $28^{\circ}C$ for 24 hrs. Maltose or fructose and peptone were selected as the best carbon and nitrogen sources, respectively. Greenhouse experiment was performed to test effectiveness of B. subtilis GG95 in the control sclerotinia rot. Drench application ($1{\times}10^8cfu/mL$, 3 times) of the bacterial culture broth to lettuce showed an effectiveness value of 88%, suggesting that B. subtilis GG95 would be a promising biocontrol agent for control of sclerotinia rot.

키워드

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피인용 문헌

  1. Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food vol.51, pp.1, 2015, https://doi.org/10.7845/kjm.2015.5004
  2. Selection of Bacillus amyloliquefaciens M27 for Biocontrol on Lettuce Sclerotinia Rot vol.43, pp.3, 2015, https://doi.org/10.4489/KJM.2015.43.3.180