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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 methods, partial fragments of the CP protein genes of three viruses of WMV, CMV and ZYMV were cloned from melon infected with virus disease in Xinjiang. Based on the single RT-PCR amplification conditions, single-factor analysis was used to optimize the multiplex RT-PCR reaction system, and 48 samples were tested by multiplex PT-PCR. The results showed that the results of multiple RT-PCR were consistent with the results of single-copy RT-PCR, indicating that the constructed multiplex RT-PCR system has a certain accuracy and stability and can be used for the rapid detection of WMV, CMV and ZYMV viruses, Pathogen identification, positive screening and other research. By analyzing the results of multiple RT-PCR in the main planting areas of melon, the distribution status and forms of the three viruses in the main planting areas of melon in Xinjiang were preliminarily obtained, which provided the basis for the prevention and control of the virus diseases, Production of virus control, field testing, epidemiological investigation and anti-virus melon breeding and other research provide the theoretical basis and basis.