Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Diversity and Antimicrobial Activity of Actinomycetes from Fecal Sample of Rhinoceros Beetle Larvae

장수풍뎅이 유충의 분변에 존재하는 방선균의 다양성 및 항균활성

  • Lee, Hye-Won (Division of Agricultural Microbiology, National Academy of Agricultural Science, RDA) ;
  • Ahn, Jae-Hyung (Division of Agricultural Microbiology, National Academy of Agricultural Science, RDA) ;
  • Kim, Minwook (Department of Biology, Kyung Hee University) ;
  • Weon, Hang-Yeon (Division of Agricultural Microbiology, National Academy of Agricultural Science, RDA) ;
  • Song, Jaekyeong (Division of Agricultural Microbiology, National Academy of Agricultural Science, RDA) ;
  • Lee, Sung-Jae (Department of Biology, Kyung Hee University) ;
  • Kim, Byung-Yong (Division of Agricultural Microbiology, National Academy of Agricultural Science, RDA)
  • 이혜원 (농촌진흥청 국립농업과학원 농업미생물과) ;
  • 안재형 (농촌진흥청 국립농업과학원 농업미생물과) ;
  • 김민욱 (경희대학교 생물학과) ;
  • 원항연 (농촌진흥청 국립농업과학원 농업미생물과) ;
  • 송재경 (농촌진흥청 국립농업과학원 농업미생물과) ;
  • 이성재 (경희대학교 생물학과) ;
  • 김병용 (농촌진흥청 국립농업과학원 농업미생물과)
  • Received : 2013.06.11
  • Accepted : 2013.06.25
  • Published : 2013.06.30
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Abstract

Actinomycetes produce diverse secondary metabolites which have the primary importance in medicine, agriculture and food production, and key to this is their ability to interact with other organisms in natural habitats. In this study, we have investigated the taxonomical and functional diversity of actinomycetes in fecal sample of rhinoceros beetle larvae (Allomyrina dichotoma L.) by using culture-dependent and -independent approaches. For the culture-independent approach, the community DNA was extracted from the sample and 16S rRNA genes of actinomycetes were amplified using actinomycetes-specific PCR primers. Thirty-seven clones were classified into 15 genera and 24 species of actinomycetes. For the culture-dependent approach, 53 strains were isolated from larval feces, of which 27 isolates were selected based on morphological characteristics. The isolates were classified into 4 genera and 14 species, and 24 isolates (89%) were identified as the genus Streptomyces. Many of the representative isolates had antimicrobial activities against plant pathogenic fungi and Gram-positive bacteria. In addition, most of the isolates (78%) showed biochemical properties to hydrolyze cellulose and casein. The results demonstrated that diverse and valuable actinomycetes could be isolated from insect fecal samples, indicating that insect guts can be rich sources for novel bioactive compounds.

방선균은 의약, 농업 및 식품 생산 등에 유용하게 사용되는 이차대사물질을 생산하는 미생물 자원으로 자연환경 내에서 많은 생물들과 긴밀한 상호작용을 유지하고 있다. 본 연구에서는 장수풍뎅이 유충의 분변에 존재하는 방선균의 다양성과 기능성을 조사하기 위해서 비배양적 접근과 배양적 방법으로 실험을 수행하였다. 먼저 시료로부터 직접 추출한 community DNA에서 방선균-특이 primer를 이용하여 방선균의 16S rRNA 유전자를 PCR로 증폭, 클로닝한 후에 각 clone에 삽입된 염기서열을 분석하였다. 총 37개의 염기서열을 얻었으며 계통분류학적 분석을 수행한 결과, 15속 24종으로 분류되었다. 아울러 53개의 방선균 균주를 장수풍뎅이 유충 분변으로부터 분리하였다. 형태학적 특징을 비교하여 최종적으로 27개의 균주를 선발하여 다양성, 항균활성 및 생화학적 특징을 검정하였다. 분리된 균주들은 4속 14종으로 분류되었으며, 24균주(89%)는 Streptomyces 속으로 분류되었다. 대다수의 균주들이 식물병원성 곰팡이와 그람양성 세균에 대해 길항 효과를 보였다. 또한 많은 균주들이 셀룰로오스와 카제인을 분해하는 생화학적 특징을 보였다. 본 연구를 통해, 장수풍뎅이 유충의 분변 시료로부터 다양한 방선균이 분리될 수 있으며, 분리된 균주들은 다양한 항균효과 및 효소활성을 지니고 있다는 것을 확인하였다. 결론적으로 본 연구는 장수풍뎅이와 같은 곤충의 장과 분변은 다양한 방선균의 서식처이며, 이곳에서 유용한 생리활성 물질을 발견할 수 있다는 것을 제시한다.

Keywords

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