基于多重RT-PCR技术检测新疆甜瓜主栽区3种病毒病及其分布

Detection and Distribution of Three Viral Diseases in Xinjiang Melon Region Based on Multiplex RT-PCR Technique

采用同源克隆的方法,从新疆感染病毒病的甜瓜中克隆获得WMV、CMV、ZYMV 3种病毒CP蛋白基因的部分片段。并基于单重RT-PCR扩增条件,采用单因素分析法,优化所获得多重RT-PCR的反应体系,利用多重PT-PCR体系对48份供试样品检测。结果表明,多重RT-PCR的检测结果与单重RT-PCR结果一致,说明构建的多重RT-PCR体系具有一定准确性和稳定性,可以用于WMV、CMV、ZYMV 3种病毒的快速检测、病源鉴定、阳性筛选等研究。通过分析甜瓜主要种植区病毒病的多重RT-PCR检测结果,初步得出新疆甜瓜主栽区3种病毒的分布现状与侵染形式,从而为病毒病的防治工作提供依据,以期对新疆瓜果生产中病毒防治,田间检测,流行病学调查以及抗病毒病甜瓜育种等研究提供相应的理论基础与依据。

Using homologous cloning method,we cloned partial fragments of coat protein gene of WMV,CMV and ZYMV from virus-infected melon in Xinjiang.the reaction system of the multiplex RT-PCR was optimized based on RT-PCR amplification conditions,using single factor analysis.Forth-eight samples were tested separately with multiple RT-PCR and single RT-PCR.The results indicated that the both test results were consistent.The optimized multiplex RT-PCR system has a certain accuracy and stability,and can be used for quick detection and positive screening of WMV（Watermelon mosaic virus）,CMV（Cucumber mosaic virus）,and ZYMV（Zucchini yellow mosaic virus）virus pathogen.The distribution status and infection incidence of three kinds of viruses was determined in mainly melon growing area of Xinjiang,which providing theoretical foundation and experimental evidence for viral diseases control,field testing,epidemiological research and melon virus-resistant breeding in Xinjiang.