四种草莓病毒SMoV、SVBV、SCV、SMYEV多重RT-PCR检测

Detection of four viruses( SMoV、SVBV、SMYEV and SCV)infecting strawberry by multiplex RT-PCR

我国草莓病毒主要有草莓斑驳病毒(SMoV)、草莓镶脉病毒(SVBV)、草毒皱缩病毒(SCV)和草莓轻型黄边病毒(SMYEV),通常发生混合侵染。本研究针对我国4种草莓病毒的外壳蛋白基因部分序列设计引物,通过优化多重RT-PCR反应条件,建立起能同时检测4种草莓病毒的多重RT-PCR检测体系。应用该体系成功对4种病毒复合侵染的草莓材料进行多重RT-PCR扩增,得到278、394、731和861bp共4条特异性条带。测序结果表明,4种病毒序列与相应的参考序列相似性均达98%以上。对草莓茎尖脱毒和叶片离体再生的试管苗样品进行检测,结果显示,该方法可以同步、快速、特异地检测出受侵染寄主植物中的4种病毒,并且灵敏度高。

Strawberry plants are frequently infected by Strawberry Mottle Virus （SMoV）,Strawberry Vein Banding Virus （SVBV） ,Strawberry Crinkle Virus（SCV） and Strawberry Mild Yellow Edge Virus（ SMYEV）, and sometimes complexly infected in China. Four pairs of specific primers were designed according to the partial coat protein gene sequences of the four viruses. A method of detecting SMoV,SVBV,SCV and SMYEV by multiplex RT-PCR was established through progressively adjusting and optimizing the RT-PCR conditions. The expected fragments of 278 bp （ SVBV）, 394 bp （SMoV） ,731 bp （SCV） and 861 bp （SMYEV） were successfully amplified by the multiplex RT-PCR system. The results verified that this system was useful for simultaneous detection of these four viruses from microplant obtained by the stem tip culture and leave regeneration.