利用siRNA高通量测序技术检测烟草病毒

摘要

：植物可以利用小干扰RNA(siRNA)机制干扰体内RNA 病毒的增殖，从而获得对该病毒的免疫能力。为了快速检测田间烟草病毒病种类和发现烟草新病毒，本文通过第二代测序技术对烟草的小RNA 进行高通量测序，利用软件velvet version 1.1.06对测得的短序列进行组装，再通过生物信息学方法将得到的重叠群进行数据库比对寻找其中的病毒序列。研究结果得出我国安徽皖南烟区烟草存在PVY、CMV、TMV 和TVBMV 等RNA 病毒。检测到3个重叠群与Potato leafroll virus 部分同源，同源性低于90%，疑似一种烟草新病毒。至此建立了检测烟草病毒和发现新病毒的一种新方法，可用于大田烟草病毒的本底调查。%Quick detection of tobacco virus strains and identification of new virus were achieved through 2nd generation sequencing technique to perform high throughput sequencing of tobacco siRNA and then screening for RNA virus sequences with bioinformatics methods. Results showed that PVY, CMV, TMV, TVBMV and other virus genomic information were detected in southern part of Anhui tobacco growing areas. Three contigs were identified as partly homologous withPotatoleafroll virus, which might be a novel tobacco virus. This research established a new strategy to determine naturally occurred RNA viruses and novel virus in tobacco, and could be applied to virus surveillance in tobacco.