Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
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PCR based Rapid Isolation of Agrobacterium vitis Strains in Korea and Their Pathogenical and Biochemical Characteristics

PCR 특이검출에 의한 국내 포도나무 혹병(Agrobacterium vitis) 균주의 신속 분리 및 병원학적, 생화학적 특성 비교

  • Kim, Jong-Kun (Graduate School of Biotechnology and Information Technology, Hankyong National University) ;
  • Lim, Sun-Hwa (Graduate School of Biotechnology and Information Technology, Hankyong National University) ;
  • Lee, Dae-Sung (Graduate School of Biotechnology and Information Technology, Hankyong National University) ;
  • Choi, Jae-Eul (College of Agriculture, Chungnam National University) ;
  • Yun, Hae-Keun (National Horticultural Research Institute, RDA) ;
  • Park, Sang-Hun (Agriculture Technology Center) ;
  • Kang, Sung-Su (Agriculture Technology Center) ;
  • Kang, Hee-Wan (Graduate School of Biotechnology and Information Technology, Hankyong National University)
  • 김종군 (한경대학교 생물환경.정보통신전문대학원) ;
  • 임선화 (한경대학교 생물환경.정보통신전문대학원) ;
  • 이대성 (한경대학교 생물환경.정보통신전문대학원) ;
  • 최재을 (충남대학교 농업생명과학대학) ;
  • 윤해근 (농촌진흥청원예연구소) ;
  • 박상헌 (천안시농업기술센터) ;
  • 강성수 (천안시농업기술센터) ;
  • 강희완 (한경대학교 생물환경.정보통신전문대학원)
  • Published : 2006.12.01
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Abstract

Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. Bacterial strains were isolated from crown galls of different grapevine varieties in grapevine farms of Kyungbuk(Kimcheon), Chungbuk(Okcheon), Chungnam(Daejon, Choenan) and Kyeonggi(Suwon, Ansung) areas in Korea from 2002 to 2005. Primer sets, Phe A and VirA, which were derived from pectate lysase gene and virA gene of Ti-plasmid in A. vitis were used to detect A. vitis strains from crown galls. PheA and VirA primers amplified DNA fragments of 0.25 kb and 0.5 kb from fifty-one bacterialstrains. They formed crown galls on grapevine variety, Kyoho, or carrot disks with variable pathgenecity It was confirmed that the biochemical characteristics of 10 bacterial strains that was strong pathogene city on grapevine were mostly in agreement with type culture strains of A. vitis, showing growth in the presence of 2% NaCl, non-production of acid from melezitose and negative response in production of 3-Ketolactose.

2002년에서 2005년 동안 경북김천, 천안, 경기지역의 포도재배지로부터 채집한 300여점의 포도 혹병반으로부터 A. vitis의 PCR 특이검출 Phe A primer를 이용하여 포도혹병균의 대량분리를 실시하여 0.25kb의 PCR 특이밴드가 증폭된 51균주가 포도 혹병균으로 선발되었다. 이 균주들을 Ti-Plasmid 유래의 Vir A primer로 적용한 결과 40균주에서 0.5kb의 특이밴드가 검출되었다. 거봉포도와 당근절편에서 다양한 양상의 병원성을 보였으며 25균주가 거봉포도에 강한 병원성을 나타내었으며, 다른 균주는 약하거나 비 병원성을 보였다. 본 연구에서 분리한 10개 균주를 대상으로한 세균의 이화학적 특성은 균주간에 약간은 달랐으나, 3-Ketolactose의 비생성, 2%NaC 첨가 배지에서의 성장, Melezitose로부터의 산을 생성하지 못하는 등 비교균주와 대체적으로 일치하는 경향을 보였다. 따라서, PheA and Vir A primer를 이용하여 Agrobacterium vitis 균주를 대량 분리할 수 있으며 신속, 정확하게 동정할 수 있을 것으로 생각되었다.

Keywords

References

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