Chen Sha, Huang Qingqing, Wu Liqi, Qian Yajuan
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, People's Republic of China.
Virol J. 2015 Oct 5;12:156. doi: 10.1186/s12985-015-0384-3.
Maize streak Reunion virus (MSRV) is a member of the Mastrevirus genus in the family Geminiviridae. Of the diverse and increasing number of mastrevirus species found so far, only Wheat dwarf virus and Sweetpotato symptomless virus 1 have been discovered in China. Recently, a novel, unbiased approach based on deep sequencing of small interfering RNAs followed by de novo assembly of siRNA, has greatly offered opportunities for plant virus identification.
Samples collected from maize leaves was deep sequencing for virus identification. Subsequently, the assay of PCR, rolling circle amplification and Southern blot were used to confirm the presence of a mastrevirus.
Maize streak Reunion virus Yunnan isolate (MSRV-[China:Yunnan 06:2014], abbreviated to MSRV-YN) was identified from maize collected from Yunnan Province, China, by small RNA deep sequencing. The complete genome of this virus was ascertained as 2,880 nucleotides long by conventional sequencing. A phylogenetic analysis showed it shared 96.3 % nucleotide sequence identity with the isolate of Maize streak Reunion virus from La Reunion Island. To our knowledge, this is the first identification of MSRV in China. Analyses of the viral derived small interfering RNAs (vsiRNAs) profile showed that the most abundant MSRV-YN vsiRNAs were 21, 22 and 24 nt long and biased for A and G at their 5' terminal residue. There was a slightly higher representation of MSRV-YN siRNAs derived from the virion-sense strand genome than the complementary-sense strand genome. Moreover, MSRV-YN vsiRNAs were not uniformly distributed along the genome, and hotspots were detected in the movement protein and coat protein-coding region.
A mastrevirus MSRV-YN collected in Yunnan Province, China, was identified by small RNA deep sequencing. This vsiRNAs profile derived from MSRV-YN was characterized, which might contribute to get an insight into the host RNA silencing defense induced by MSRV-YN, and provide guidelines on designing antiviral strategies using RNAi against MSRV-YN.
玉米条纹团聚病毒（MSRV）是双生病毒科玉米褪绿斑驳病毒属的成员。在迄今发现的种类多样且数量不断增加的玉米褪绿斑驳病毒物种中，中国仅发现了小麦矮缩病毒和甘薯无症状病毒1。最近，一种基于小干扰RNA深度测序并随后对siRNA进行从头组装的新型无偏方法，为植物病毒鉴定提供了极大的机会。
对从玉米叶片采集的样本进行深度测序以鉴定病毒。随后，使用PCR检测、滚环扩增和Southern杂交来确认玉米褪绿斑驳病毒的存在。
通过小RNA深度测序，从中国云南省采集的玉米中鉴定出玉米条纹团聚病毒云南分离株（MSRV-[中国：云南06:2014]，简称为MSRV-YN）。通过常规测序确定该病毒的完整基因组长度为2880个核苷酸。系统发育分析表明，它与来自留尼汪岛的玉米条纹团聚病毒分离株具有96.3%的核苷酸序列同一性。据我们所知，这是MSRV在中国的首次鉴定。对病毒衍生的小干扰RNA（vsiRNA）谱的分析表明，最丰富的MSRV-YN vsiRNA长度为21、22和24 nt，并且在其5'末端残基处偏向于A和G。来自病毒链基因组的MSRV-YN siRNA的代表性略高于互补链基因组。此外，MSRV-YN vsiRNA并非均匀分布在基因组中，在运动蛋白和外壳蛋白编码区域检测到热点。
通过小RNA深度测序鉴定了在中国云南省采集的一种玉米褪绿斑驳病毒MSRV-YN。对源自MSRV-YN的这种vsiRNA谱进行了表征，这可能有助于深入了解由MSRV-YN诱导的宿主RNA沉默防御，并为使用RNAi针对MSRV-YN设计抗病毒策略提供指导。
