종자전염검역바이러스 3종(CRLV, SpLV 및 WClMV)을 검정하기 위한 RT-PCR 및 Nested PCR 프라이머 개발
Development of Nucleotide Primers for Dignostic RT-PCR and Nested PCR Detection of Three Seed-transmitted Viruses (CRLV, SpLV and WClMV) in Quarantine
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초록
종자의 수입 시, 검역관련 종자전염바이러스는 가장 문제가 되는 식물병이다. 본 연구에서 PCR 검역체계가 보고되지 않은 3종의 종자전염바이러스, Cherry rasp leaf virus (CRLV), Spinach latent virus(SpLV) 및 White clover mosaic virus (WClMV)를 검출하기 위하여 reverse transcriptionpolymerase chain reaction (RT-PCR)과 nested polymerase chain reaction (nested PCR) 방법을 도입하였다. 각각의 바이러스별로 2 세트의 RT-PCR primer가 선발되었으며, 증폭산물에서 더욱 높은 감도로 검출 할 수 있는 nested PCR primer set를 개발하였다. 본 연구에서 사용한 RT-PCR과 nestedPCR 방법은 종자로부터 CRLV, SpLV 및 WClMV를 검역하는 고효율적 진단시스템으로 제공될 것이다.
Abstract
Seed-transmitted viruses regarding quarantine are the most problematic plant diseases in cropseed industry. In this study, three seed-transmitted viruses [Cherry rasp leaf virus (CRLV),Spinach latent virus (SpLV) and White clover mosaic virus (WClMV)] that have not previouslybeen studied for PCR diagnostic system in Korean quarantine were targeted for the detection. For successful virus detection, we employed a diagnostic technique based on reverse transcriptionpolymerase chain reaction (RT-PCR) and nested polymerase chain reaction (nested PCR) methods. Two RT-PCR primer sets for each virus were finally selected for the diagnosis. Nested primersets developed in the present study were shown to be highly sensitive in detection andverification of the target viruses. Overall, RT-PCR and nested PCR were proven to be a usefuldiagnostic technique for the detection of CRLV, SpLV and WClMV in quarantine.
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