侵染四川辣椒的番茄斑萎病毒和辣椒轻斑驳病毒的小RNA测序检测及鉴定

 调查发现四川省汶川县当地辣椒的病毒病严重且症状多样,病样粗提液摩擦接种辣椒、矮牵牛和三生烟,出现辣椒系统性花叶焦枯和茎尖坏死,指示植物表现局部枯斑。对3个不同症状的病果进行sRNA深度测序鉴定,发现均含有番茄斑萎病毒(Tomato spotted wilt virus,TSWV)和辣椒轻斑驳病毒(Pepper mild mottle virus,PMMoV)。通过RT-PCR技术进行验证,结果显示所有病样的果皮和部分新鲜种子以及回接寄主的病叶均检测到TSWV和PMMoV,表明该地辣椒病毒病是由TSWV和PMMoV复合侵染引起。这是TSWV侵染四川辣椒的首次报道。分别基于TSWV N基因序列和PMMoV CP基因序列构建系统发育树,汶川分离物TSWV-WC(MK468469)与贵州分离物(KP684518)亲缘关系最近,PMMoV-WC(MK408614)与北京分离物(AY859497)亲缘关系最近。推测该地辣椒病毒病可能与品种引进有关。

Identification and detection of Tomato spotted wilt virus and Pepper mild mottle virus infecting peppers in Sichuan using small RNA sequencing

Pepper virus disease showing severe and multiple symptoms broken out in Wenchuan county of Sichuan province. Systemic mosaic,withered leaves and stem tip necrosis were observed in pepper and partial necrosis in Petunia hybrida and Nicotiana tabacum cv. Samsun NN through the method of mechanical inoculation.We found that Tomato spotted wilt virus and Pepper mild mottle virus were presented in all samples tested by small RNA deep sequencing. The two viruses were detected in all peels and some seeds of the tested samples and re-inoculation of the host followed by RT-PCR further verified that viral disease in this area was caused by co-infection of these two viruses. Pepper-infecting TSWV was reported for the first time from Sichuan region.The phylogenetic tree constructed based on TSWV N gene and PMMoV CP gene sequence respectively indicated that TSWV-WC(MK468469) was more closely related to Guizhou isolate(KP684518), whereas PMMoV-WC(MK408614) remained closest to Beijing isolate(AY859497). We supposed that outbreak of pepper virus disease in the region was related to seeds imported from other regions.