4种大豆种传病毒多重RT-PCR检测

Detection of four soybean seed transmitted viruses by multiplex RT-PCR

进境大豆携带的病毒主要有菜豆荚斑驳病毒(BPMV)、烟草环斑病毒(TRSV)、烟草条纹病毒(TSV)和南方菜豆花叶病毒(SBMV)等,均为大豆种传病毒,可随大豆种子实现远距离传播。本研究对这4种大豆病毒的外壳蛋白基因部分序列设计引物,并通过优化引物、模板浓度和扩增参数,在一个体系中成功对4种病毒进行了多重RT-PCR扩增,得到307bp、206bp7、17bp5、18bp共4条特异性条带,建立了能同时检测BPMV、TRSV、TSV和SBMV的多重RT-PCR检测体系。该方法快速、灵敏、简便,同时特异性强,在出入境检疫中具有广泛的应用前景。

Imported soybean are frequently infected by bean pod mottle virus（BPMV）,tobacco ringspot virus（TRSV）,tobacco streak virus（TSV） and Southern bean mosaic virus（SBMV）.All of them can be transmitted through soybean seeds.Four pairs of specific primers were designed according to the partial coat protein gene sequences of the four viruses.By optimization of primer,template concentration and amplification parameters,the expected fragments of 307bp,206bp,717bp and 518bp were successfully amplified by the multiplex RT-PCR.This approach suggested a simple,sensitive,rapid and more specific detection of these four viruses from soybean samples for the entry-exit quarantine.