五种烟草病毒TMV、CMV、TEV、PVY及TVBMV的多重RT-PCR同步检测

Simultaneous detection of five viruses (TMV,CMV,TEV,PVY and TVBMV) infecting tobacco by multiplex RT-PCR

我国烟草病毒主要有烟草花叶病毒(TMV)、黄瓜花叶病毒(CMV)、烟草蚀纹病毒(TEV)、马铃薯Y病毒(PVY)和烟草脉带花叶病毒(TVBMV),通常发生复合侵染。本研究对我国5种烟草病毒的外壳蛋白基因部分序列设计引物,通过优化引物和模板浓度,摸索扩增参数,在一个体系中成功对5种病毒复合侵染的烟草材料进行多重RT-PCR扩增,得到237、273、347、456和547 bp共5条特异性条带,建立了能同时检测TMV、CMV、TEV、PVY和TVBMV的多重RT-PCR检测体系。对田间样品检测结果证明,多重RT-PCR体系能够同时检测5种病毒,并且灵敏度高。

Tobacco plants are frequently infected by Tobacco mosaic virus（TMV）,Cucumber mosaic virus（CMV）,Tobacco etch virus（TEV）,Potato virus Y（PVY） and Tobacco vein banding mosaic virus（TVBMV）,and sometimes complexly infected in China.Five pairs of specific primers were designed according to the partial coat protein gene sequences of the five viruses.By optimization of primer,template concentration and amplification parameters,the expected fragments of 237 bp（TMV）,273 bp（CMV）,347 bp（TEV）,456 bp（PVY） and 547 bp（TVBMV） were successfully amplified by the multiplex RT-PCR system.The results verified that this system was useful for simultaneous detection of these five viruses from field samples.