利用小RNA深度测序和组装技术鉴定紫藤花叶病病原  

Detection of viruses infecting Wisteria sinensis by deep sequencing and assembly of small RNA

作　　者：苏秀[1,2] 徐毅[1] 陈莎[1] 傅帅[1] 钱亚娟[1] 张立钦[2] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,杭州310058 [2]浙江农林大学亚热带森林培育国家重点实验室培育基地,临安311300

出　　处：《植物病理学报》2015年第1期88-92,共5页Acta Phytopathologica Sinica

基　　金：质检公益性行业科研专项项目(201310068);浙江省重中之重林学一级学科开放基金项目(KF201330);浙江农林大学科研发展基金项目(2013FK019)

摘　　要：RNA沉默（RNA silencing）是一种在真核生物体内普遍保守的基于核酸序列特异性抑制基因表达的调控机制。2009年Kreuze等发现病毒特异的小RNA（small RNA，sRNA）在序列上是重叠的，因此推测通过深度测序技术获得的大量sRNA序列能用来组装病毒的基因组并用来鉴定和发现新病毒。利用sRNA深度测序技术已在作物和昆虫上鉴定发现多种病毒。但在木本植物上还未见报道。Plant defense against viruses through small RNA （sRNA） mediated RNA interference mechanism. Analysis of virus derived sRNA profiles in plant can be applied for de novo assembly of virus genomes and virus identification. In this study, suspected virus-infected Wisteria sinensis samples collected from Zijingang Campus of Zhejiang University were used for sRNA library construction and deep sequencing. After assembly of total sRNAs, it was found that W. sinensis leaves were infected by Wisteria vein mosaic virus （WVMV）. The library generated 18.9 million sRNA reads, of which 0. 32 million were WVMV-derived sRNAs. Using de novo assembly, 23.3% of full length genome nucleotide sequence of a previously reported potyvirus WVMV was obtained. To confirm the existence of WVMV in the samples, WVMV coat protein （CP） gene sequence was obtained by RT-PCR, and ensured by Sanger sequencing. Taken together, the data suggest that sRNA deep sequencing technology is an efficient and powerful genetic tool for virus identification in woody plants.

关 键 词：技术鉴定 测序技术 小RNA 组装 利用 花叶病 序列特异性 病原 

分 类 号：S858.31[农业科学—临床兽医学]