Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
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Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts

잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성

  • Lee, Hyeok-Geun (Division of Bio-resources Technology, Kangwon National University) ;
  • Kim, Sung-Ryul (Division of Bio-resources Technology, Kangwon National University) ;
  • Jeon, Yong-Woon (Division of Bio-resources Technology, Kangwon National University) ;
  • Kwon, Soon-Bae (Kangwondo Agricultural Research and Extension Services) ;
  • Ryu, Ki-Hyun (Division of Environmental and Life Sciences, Seoul Women's University) ;
  • Choi, Jang-Kyung (Division of Bio-resources Technology, Kangwon National University)
  • 이혁근 (강원대학교 생물자원공학부) ;
  • 김성률 (강원대학교 생물자원공학부) ;
  • 전용운 (강원대학교 생물자원공학부) ;
  • 권순배 (강원도농업기술원) ;
  • 류기현 (서울여자대학교 환경생명과학부) ;
  • 최장경 (강원대학교 생물자원공학부)
  • Published : 2008.04.01
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Abstract

Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.

산박하(Isodon inflexus)의 Is-CMV, 깽깽이풀(Jeffersonia dubia)의 Jd-CMV 및 파리풀(Phryma leptostachya var. asiatica)의 Pla-CMV 등, 3종의 잡초에서 분리한 Cucumber mosaic virus(CMV)를 공시하여, 기주반응 실험, dsRNA 분석, 혈청학적 성질조사, RT-PCR및 RFLP등의 실험을 통하여 각 바이러스를 동정하고, 특성을 구분하였다. 잡초로부터 분리 한 3종의 CMV는 Nicotiana benthamiana, N. tabacum cv. Xanthi nc, N. glutinosa, Cucubita pepo cv. Black Beauty에는 모두 유사한 모자이크 병징을 발현하였으며, Chenopodium amaranticolr와 Vigna unguiculata cv. Kurotanesanzaku에서는 국부 괴사병반이 발현되었다. 한편 고추(Capsicum anmuum cv. Chungyang)에서는 Jd-CMV와 Pla-CMV는 전형적인 모자이크 증상을 발현하였으나, Is-CMV는 병징이 나타나지 않고 무병징으로 감염되는 특성을 보였다. Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana로부터 추출한 dsRNA는 모두 약 3.4, 3.2, 2.1 및 1.0kbp의 분자크기를 갖는 4종의 dsRNA 밴드가 검출되었으며, 대조로 이용한 Fny-CMV의 dsRNA 패턴과 같았다. Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana의 즙액을 항원으로 이용하여 Fny-CMV의 항혈청과 한천겔이중확산법으로 조사한 혈청학적 실험 결과는 모든 항원이 한 종의 뚜렷한 침강선을 형성 하였으며, Fny-CMV의 항원에 의해서 형성된 침강선과 융합함으로서 서브그룹 I에 속하는 계통들로 판단되었다. 또한 Is-CMV, Jd-CMV 및 Pla-CMV에 감염된 N. benthamiana로부터 추출한 RNA를 이용하여 CMV-specific 프라이머를 이용한 외피단백질유전자를 포함하는 RNA3의 3' 영역 에 대한 RT-PCR을 실시한 결과, Fny-CMV와 마찬가지로 약 950bp 크기의 cDNA가 증폭되었다. 증폭된 각각의 cDNA를 EcoRI으로 처리하였을 때에는 절단되지 않았으며, HindIII, MspI, SalI 그리고 XhoI에서는 2개의 절편으로 절단되었다. 이와 같은 결과는 Fny-CMV의 절단패턴과 일치하는 것으로 Is-CMV, Jd-CMV 그리고 Pla-CMV는 서브그룹 IA에 속하는 계통으로 확인되었다. 이들 3종의 잡초로부터 CMV가 분리 동정된 것은 이 논문이 처음이다.

Keywords

References

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