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Characterization of Tobacco rattle virus(TRV-K) isolated in Korea

한국에서 분리한 Tobacco rattle virus(TRV-K)의 특성

  • 신혜영 (영남대학교 이과대학 생물학과) ;
  • 구봉진 (영남대학교 이과대학 생물학과) ;
  • 강상구 (영남대학교 생명공학연구소) ;
  • 장무웅 (영남대학교 이과대학 생물학과) ;
  • 류기현 (서울여자대학교 환경생명과학부 원예학과)
  • Published : 2002.12.01

Abstract

Tobacco rattle virus(TRV) was detected from Gladiolus hybridus, Crocus spp. and Narcissus spp. leaves show-ing notched or stripe on the leaf and malformation symptoms collected from Daegu and Kyungbuk province by electron microscopy (EM), immunosorbent electron microscopy (ISEM) and host range study. Direct negative staining method by EM showed rigid rod long particles 170~200$\times$22 nm and rigid rod short particles 40~114$\times$22 m. TRV-K isolated from G. hybridus propagated with Nicotiana tabacum. TRV coat protein(CP) gene was amplified using specific oligonucleotide primer by RT-PCR. Sequence analysis of amplified CP gene showed 99.5% nucleotide similarity to TRV-ORY.

대구시와 경상북도 지역에서 Tobacco rattle virus(TRV) 감염에 의한 증상으로 보이는 글라디올러스(Gladiolus hybridus), 크로커스(Crocus spp.), 수선화(Narcissus spp.)를 채집하였다. 톱니무늬(notch), 줄무늬(stripe), 변형 (malformation)등의 증상의 잎 조직을 시료로 하여 direct negative staining method(DN) 및 immunosorbent electron microscopy(ISEM)법에 의해 투과전자현미경으로 바이러스 입자를 관찰한 결과, 각각의 시료에서 긴 입자는 170~200 X 22nm, 짧은 입자는 40~114$\times$22nm 크기의 막대모양 입자가 다수 확인되었다. 글라디올러스에서 분리한 tobacco rattle virus(TRV-K)를 담배(Nicotiana tabacum)에 즙액 접종, 증식시킨 후, 정제하여 항혈청을 제작하였다. TRV-K 의 coat protein(CP)유전자를 특이적인 올리고뉴클레오타 이드 프라이머를 이용하여 역전사-중합효소연쇄반응(RT-PCR)으로 증폭시킨 후에 염기서열분석법으로 확인하였다. 그 결과 TRV-ORY의 CP와 99.5%의 상동성을 나타냈다.

Keywords

References

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