利用小RNA深度测序技术鉴定海南南瓜病毒种类  

Identification of Viruses from Pumpkin in Hainan by Small RNA Deep Sequencing Technology

作　　者：车海彦[1] 曹学仁[1] 刘勇 罗大全[1] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所农业部热带作物有害生物综合治理重点实验室海南省热带农业有害生物监测与控制重点实验,海南海口571101 [2]湖南省植物保护研究所,湖南长沙410125

出　　处：《热带作物学报》2017年第11期2106-2111,共6页Chinese Journal of Tropical Crops

基　　金：国家公益性行业(农业)科研专项(No.201303028)

摘　　要：2016年在海南乐东、昌江、澄迈、海口、文昌、儋州等地采集疑似被病毒病感染的南瓜叶片样品6个,将样品合并,采用小RNA深度测序技术对上述混合样本的病毒种类进行鉴定,发现样本中含有黄瓜花叶病毒(Cucumber mosaic virus,CMV)、番木瓜环斑病毒(Papara ringspot virus,PRSV)、黄瓜绿斑驳病毒(Cucumber green mottle mosaic virus,CGMMV)和中国南瓜曲叶病毒(Squash leaf curl China virus,SLCCNV),通过PCR方法进一步验证了深度测序结果的准确性。SLCCNV在4个样品中存在,P-CJ分离物的DNA-A和DNA-B组分全长分别为2 735 bp(登录号为MF062251)和2 722 bp(登录号为MF062252)。DNA-A和DNA-B组分均与SLCCNV分离物的同源性最高,同源性分别为89.2%~99.3%和79.1%~98.8%;P-CJ分离物与从中国植物样品中分离到的SLCCNV聚在同一进化分支上。上述研究结果表明,P-CJ是SLCCNV的一个分离物。In 2016, 6 suspected virus-infected pumpkin leaf samples were collected from Ledong county, Changjiang county, Chengmai county, Haikou city, Wenchang county and Danzhou city, Hainan province. The above samples were pooled and analyze by small RNA deep sequencing. Cucumber mosaic virus（ CMV）, Papaya ringspot virus（ PRSV）, Cucumber green mottle mosaic virus（ CGMMV） and Squash leaf curl China virus（ SLCCNV） were detected from the pooled sample. Viruses identified by small RNA sequencing were validated by virus-specific PCR from above 6 samples. SLCCNV was detected from 4 samples, the virus isolate P-CJ was selected for further sequence analysis. The complete sequence of the DNA-A and DNA-B was determined to be 2 735 bp（MF062251）and 2 722 bp（MF062252） respectively. The DNA-A was most closely related to isolates of SLCCNV with 89.2%-99.3% nucleotide sequence identities. The DNA-B was most closely related to isolates of SLCCNV with 79.1%-98.8%nucleotide sequence identities. Phylogenetic analysis revealed that P-CJ isolate clustered with isolates of SLCCNV from China. These results confirmed this virus was an isolate of SLCCNV.

关 键 词：小RNA深度测序 海南 南瓜病毒病 

分 类 号：S432.1[农业科学—植物病理学]