The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Detection of Pseudomonas tolaasii Causing Brown Blotch Disease of Mushroom with Species-specific DNA Probe

종 특이 DNA probe를 이용한 버섯 세균성 갈반병 병원균(Pseudomonas tolaasii)의 검출

  • Kwon, Soon-Wo (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Go, Seung-Joo (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Cheun, Meung-Sook (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Kang, Hee-Wan (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Oh, Se-Jong (Division of Applied Microbiology, NIAST, RDA) ;
  • Chang, Who-Bong (Chungbuk Agricultural Research and Extension Services) ;
  • Ryu, Jin-Chang (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA)
  • 권순우 (농업과학기술원 분자유전과) ;
  • 고승주 (농업과학기술원 분자유전과) ;
  • 전명숙 (농업과학기술원 분자유전과) ;
  • 강희완 (농업과학기술원 분자유전과) ;
  • 오세종 (농업과학기술원 응용미생물과) ;
  • 장후봉 (충북 농업기술원) ;
  • 류진창 (농업과학기술원 분자유전과)
  • Published : 1999.04.30
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Abstract

This study was carried out to develop the molecular marker for the detection of Pseudomonas tolaasii, a causative agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). When several primers designed from repetitive sequences and pectin lyase genes of bacteria were used to produce DNA polymorphism from different Pseudomonas spp. isolated from edible mushrooms, PEU1 primer derived from pectin lyase gene produced polymorphic bands differentiating P. tolaasii strains from other Pseudomonas species. Two bands, 1.0kb and 0.4kb, found commonly in 6 isolates of P. tolaasii were cloned into pGEM-T vector which were designated as pPTOP1 and pPTOP2, respectively, to use as probe. The 0.4 kb insert of pPTOP2 hybridized to only 6 isolates of P. tolaasii, but did not to the other Pseudomonas species. As few as $1.5{\times}10^3$ colony forming unit (cfu) of P. tolaasii could be detected by dot blot hybridization with the cloned 0.4kb DNA in pPTOP2.

본 연구는 느타리버섯 갈반병 병원균인 pseudomonas tolaasii의 진단을 위한 분자 marker를 개발하기 위해 수행되었다. 세균의 반복염기서열과 펙틴 분해효소 유전자로부터 제작된 여러개의 primer들을 이용해 식용버섯으로부터 분리된 Pseudomonas종들로부터 DNA 다형성을 유도한 바펙틴 분해 효소로부터 제작된 PEU1 primer는 다른 Pseudomonas종들로부터 P. tolaasii를 구분시키는 다형성 밴드를 생성하였다. P. tolaasii 6균주에 공통적으로 나타나는 1.0kb와 0.4kb의 두 가지 밴드를 pGEM-T에 클로닝하고 이들을 각기 pPTOP1과 pPTOP2로 명명하고 probe로 이용하였다. 0.4kb 크기의 pPTOP2의 삽입 DNA는 P. tolaasii 6균주와 hybridization이 이루어진 반면 다른 Pseudomonas종과는 반응하지 않았다. 0.4kb크기의 pPTOP2의 삽입 DNA를 probe로 이용해 dot blot hybridization한 결과 P. tolaasii는 $1.5{\times}10^3\;cfu$까지 검출이 가능함을 확인하였다.

Keywords

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