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Development of PCR Diagnosis System for Plant Quarantine Seed-borne Wheat Streak Mosaic Virus

식물검역 종자전염 Wheat Streak Mosaic Virus의 PCR 검사시스템 개발

  • Lee, Siwon (Plant Quarantine Technology Center, Animal and Plant Quarantine Agency) ;
  • Kang, Eun-Ha (Plant Quarantine Technology Center, Animal and Plant Quarantine Agency) ;
  • Chu, Yeon-Mee (Experiment & Analysis Division, Yeongnam Regional Office, Animal and Plant Quarantine Agency) ;
  • Shin, Yong-Gil (Plant Quarantine Technology Center, Animal and Plant Quarantine Agency) ;
  • Ahn, Tae-Young (Department of Microbiology, Dankook University)
  • 이시원 (농림축산검역본부 식물검역기술개발센터) ;
  • 강은하 (농림축산검역본부 식물검역기술개발센터) ;
  • 추연미 (농림축산검역본부 영남지역본부 시험분석과) ;
  • 신용길 (농림축산검역본부 식물검역기술개발센터) ;
  • 안태영 (단국대학교 미생물학과)
  • Received : 2013.02.15
  • Accepted : 2013.04.29
  • Published : 2013.06.30

Abstract

Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in Potyviridae, severely impacts wheat and corn seed worldwide, but has yet to be detected in Korea, and hence, every effort should be made to prevent its introduction. To prevent WSMV from entering the country, it is necessary to prepare a specific, sensitive, simple, and fast detection method for routine application to plant quarantine procedures. For this reason, a two-step diagnosis system consisting of RT-PCR and nested PCR is being used for WSMV detection. In addition, a novel positive control was developed for use with the system. WSMV has been detected in seed sweet corn from Japan and seed wheat from USA by a two-step diagnosis system, the details of which are described in this study. After sequence analysis, similarities of 80.6 and 100.0% with other isolates were determined by BLAST. They showed the same topology, which was classified as 4 genotypes by various phylogenetic trees, using a poly protein encoding sequence amplification. In this analysis, WSMV-JSweet-corn2868 (JX845574) is classified as clade B, while WSMV-Uwheat1944-1 (KC754959) and WSMV-Uwheat1944-2 (KC754960) belong to clade D.

Wheat streak mosaic virus (WSMV)는 Potyviridae 과 Tritimovirus 속으로 분류되는 식물병원성 바이러스로 밀과 옥수수 종자 등에 많은 피해를 주고 있다. 본 연구에서는 관리급 식물검역 종자전염바이러스인 WSMV를 신속하고 정확하게 진단하기 위한 2단계 PCR 시스템을 개발하였으며, 실험실 오염의 여부를 확인 할 수 있는 새로운 양성 대조구를 제작하였다. 본 연구에서 개발된 PCR 시스템으로 일본에서 수입한 스위트콘 종자와 미국에서 수입한 밀 종자의 검역과정에서 WSMV-JSweet-corn2868, WSMVUwheat1944-1 및 WSMV-Uwheat1944-2를 검출하였다. 검출된 증폭 산물은 다른 WSMV의 isolates와 80.60-100.00%의 유사성을 보였다. WSMV는 계통수에서 4가지 genotype이 확인되며, 이번 연구에서 검출한 isolate WSMV-JSweet-corn2868 (JX845574)은 Clade B로 분류되었고, isolate WSMV-Uwheat1944-1 (KC754959)과 WSMV-Uwheat1944-2 (KC754960)는 Clade D로 분류되었다.

Keywords

Acknowledgement

Supported by : 농림축산검역본부

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