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담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성

Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

  • 김다아 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김진수 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 길미라 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 백승경 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 최수연 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김대용 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 윤용남 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 황인천 (㈜경농 중앙연구소) ;
  • 유용만 (충남대학교 농업생명과학대학 응용생물학과)
  • Kim, Da-A (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kim, Jin-Su (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kil, Mi-Ra (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Paek, Seung-Kyoung (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Choi, Su-Yeon (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Jin, Da-Yong (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Youn, Young-Nam (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Hwang, In-Cheon (Central Research Institute, Kyung Nong Co.) ;
  • Yu, Yong-Man (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
  • 발행 : 2008.03.30

초록

국내 토양으로부터 분리한 Bacillus thuringiensis 균주에서 담배거세미나방(Spodoptera litura)에 선택적으로 살충효과를 나타내는 새로운 균주를 선발하였다. 이 균주의 결정성 독소단백질은 위상차 현미경과 주사전자현미경으로 관찰한 결과 전형적인 이중피라미드 형태이며 H serotype으로 동정한 결과 Bacillus thuringiensis subsp. CAB 109로 동정되었다. 난방제 해충인 담배거세미나방 3령충에 대해 B. t. subsp. aizawai CAB 109균주외 분리된 다른 3균주의 생물활성을 비교 검토하였다. 살충활성 결과에서 B. t. subsp. aizawai CAB 109 균주가 $1.3{\times}10^7$ (cfu/ml)에서 100% 사망률로 다른 균주에 비해 높은 활성을 나타내었다. 담배거세미나방은 배추좀나방과 다르게 7일 이상 경과한 뒤에야 100% 사충률을 나타내었다. 한편, 담배거세미나방 2령, 3령, 4령충에 대한 살충활성의 검정에서 B. t. aizawai subsp. CAB 109 균주의 $LD_{50}$값이 각각 $9.78{\times}10^5,\;6.87{\times}10^6,\;1.83{\times}10^7$ (cfu/ml)으로 살충활성을 나타내었다. B. t.를 섭식한 담배거세미나방 유충의 사망 현상은 지효성이며 대조군보다 무게가 $6{\sim}7$배정도 적으며 다음 령기로 성장하지 못하고 죽었다. 또한 B. t. subsp. aizawai CAB 109 균주의 SDS-PAGE에서의 단백질 패턴과 trypsin을 처리한 단백질의 결과에서 기존의 B. t. subsp. aizawai와 CAB 109균주와의 차이점은 발견할 수 없었다.

Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.

키워드

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