云南农业大学学报（自然科学）2017，Vol.32Issue(2)：197-203,7.DOI:10.16211/j.issn.1004-390X(n).2017.02.002
以TMV为模式建立基于宏基因组学的植物病毒检测方法
Metagenomics-based Plant Virus Detection Method with TMV Model
摘要
Abstract
The tobacco leaves infected with Tobacco mosaic virus (TMV) was positive control,and the dsRNAs were extracted and the sequences analysis were performed by random primers amplification and high throughput sequencing.The PCR products of metagenomic sequencing results showed that the detection rate of TMV was 99.84％,and statistical results of classification and abundance of genus level display Tobamovirus accounted for 99.913％.The experiments proved that the method of dsRNA extraction method of metagenomic sequencing to plant virus were established successfully.According to the method of extracting virus like particles from tobacco infected by to TMV,then extracted RNA from the virus like particles,RT-PCR was conducted using TMV specific primers,and the results showed that the TMV sequence homology was 99％.The results proved that the method of nucleic acid of virus like particles extraction was established successfully and met the requirements of metagenomic analysis.The dsRNA and virus DNA of the like particles of 40 viral samples were constructed by this study.The nucleic acid of the all the samples had good quality.20 kinds of plant viruses were detected from those samples.This study lays a foundation for the detection of plant virus populations and ecological adaptation in agricultural cultivation and non farming areas.关键词
宏基因组学/植物病毒/TMV/双链RNA/病毒类似颗粒/检测/方法Key words
metagenomic/plant viruses/TMV/dsRNA/virus-like particles/detection/method分类
生物科学引用本文复制引用
刘悦,孔宝华,王连春,王崇德,吴德喜,谢琼,甘晓帆..以TMV为模式建立基于宏基因组学的植物病毒检测方法[J].云南农业大学学报（自然科学）,2017,32(2):197-203,7.基金项目
国家自然科学基金项目(31470018). （31470018）
