한국응용곤충학회지 (Korean journal of applied entomology)

  • 제42권1호
  • /
  • Pages.65-70
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  • 2003
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  • 1225-0171(pISSN)
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  • 2287-545X(eISSN)
한국응용곤충학회 (Korean Society of Applied Entomology)
권호 표지

흰점박이꽃무지로부터 Metarhizium속 사상균의 분리 및 ribosomal DNA 염기서열에 의한 동정

Identification of Metarhizium sp. Isolated from Protaetia brevitarsis seulensis (Kolbe) Using Ribosomal DNA Sequence

  • 최지영 (농촌진흥청 농업과학기술원 잠사곤충부) ;
  • 김철학 (강원대학교 농업생명과학대학) ;
  • 제연호 (서울대학교 농생명공학부) ;
  • 최영철 (농촌진흥청 농업과학기술원 잠사곤충부) ;
  • 김종길 (농촌진흥청 농업과학기술원 잠사곤충부) ;
  • 박규택 (강원대학교 농업생명과학대학) ;
  • 김근영 (농촌진흥청 농업과학기술원 잠사곤충부)
  • 발행 : 2003.03.01
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초록

곤충자원의 대량사육을 위한 병 발생 예방과 해충의 효과적인 방제를 위하여 흰점박이꽃무지 이병충으로부터 곤충병원 사상균을 분리하였다. 전자현미경 관찰 결과 분리균주 KMA-1은 Metharizium속의 전형적인 쇠사슬형의 분생자를 paliside-like masse에 형성하였다. 따라서, 정확한 동정을 위하여 28S rRNA와 ITS염기 서열을 바탕으로 제작한 특이 프라이머쌍을 사용하여 PCR 반응을 수행하였다. 각각의 프라이머쌍을 사용한 PCR반응으로부터 특이 밴드가 검출되었으며 이 증폭 산물들의 염기 서열을 결정, 비교하였다. 분리 균주 KMA-1의 PCR산물인 28S rRNA와 ITS DNA염기서열을 GenBank데이터베이스에 등록된 염기서열 정보와의 상동성을 검색한 결과, 모두 Metarhizium anisopliae와 가장 높은 서열 상동성을 보였다. 이상의 결과로서 본 실험에서 분리 명명된 KMA-1는 M. anisopliae로 동정되었다.

For the purpose of the protection of beneficial insects from pathogens and the development of control agent against pests, a strain of Metarhizium sp. was isolated from the infected Protaetia brevitarsis seulensis larvae in Korea. Under the scanning electron microscope, the isolate, Metarhizium sp. KMA-1, showed distinct formation of conidia on the palisade-like masse which were comprised of elongate chains and this shape is a typical feature of Metarhizium species. PCR techniques were used to identify the isolate and the primers used were designed on the basis of two kinds of rRNAs sequences, 28S rRNA and internal transcribed spacer(ITS). The specific PCR products from each primer set were amplified and the DNA sequences were determined for the similarity comparison. Sequence alignment of these fragments using GenBank database resulted in the highest homology similarity between the isolate Metarhizium sp. KMA-1 and M. anisopliae. From these results, the isolate Metarhizium sp. KMA-1 in this study was identified as M. anisopliae.

키워드

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