双重DPO-RT-PCR检测南方菜豆花叶病毒及烟草环斑病毒  

Dual Priming Oligonucleotide System for Duplex Detection of Southern Bean Mosaic Virus and Tobacco Ring Spot Virus

作　　者：魏霜 袁俊杰 杨卓瑜 马新华 卢乃会 陈文 渭婷玉 龙阳 WEI Shuang;YUAN Junjie;YANG Zhuo-yu;MA Xin-hua;LU Nai-hui;CHEN Wen;WEI Ting-yu;LONG Yang(Guangzhou Customs Technology Center,Guangzhou 510000,China;Zhanjiang Customs Technology Center,Zhanjiang 524000,China;Zhanjiang Customs,Zhanjang 524000,China;Xining Custonns Technology Center,Xining 810000,China)

机构地区：[1]广州海关技术中心,广东广州510000 [2]湛江海关技术中心,广东湛江524000 [3]湛江海关,广东湛江524000 [4]西宁海关技术中心,青海西宁810000

出　　处：《大豆科学》2021年第2期265-269,共5页Soybean Science

基　　金：国家重点研发计划(2018YFC0809200);海关总署科研项目(2019HK052,2020HK147)。

摘　　要：南方菜豆花叶病毒及烟草环斑病毒是中国进境植物检疫性有害生物,属于大豆种传病害,为提高在进口大豆中两种病毒的检出率,本研究针对SBMV及TRSV基因组中的保守序列,分别设计特异性DPO引物,建立了一种快速检测这两种植物病毒的双重DPO-RT-PCR方法,并对该方法的特异性、灵敏度及退火温度敏感性等性能进行评价。结果表明:该方法能准确地从10种植物病毒中鉴定出SBMV及TRSV,表现出良好的特异性,且检测灵敏度可达0.02 ng·μL^(-1),退火温度为在45~65℃时该方法检测结果无明显差异,表现出对退火温度不敏感的特性。研究结果为进出境植物病毒的高通量检测提供了新的思路。Southern bean mosaic virus(SBMV) and tobacco ring spot virus(TRSV) are quarantine pests in China, belonging to soybean seed-borne diseases. In order to improve the detection rate of these two viruses in imported soybeans, we designed two pairs of DPO(dual priming oligonucleotide) primers based on specific sequence of SBMV and TRSV in this research, and developed a duplex DPO-RT-PCR method for the detection of SBMV and TRSV. The method has been evaluated for its specificity, sensitivity and annealing temperature sensitivity. The result showed that this method could accurately identify SBMV and TRSV from 10 kinds of plant viruses, showing good specificity and detection sensitivity up to 0.02 ng·L^(-1). The detection results of this method had no significant difference at the annealing temperature of 45-65 ℃, showing the insensitivity characteristics to annealing temperature. The study provides a new method for high-throughput detection of plant pathogens.

关 键 词：南方菜豆花叶病毒 烟草环斑病毒 进口大豆 种传病毒 检测 DPO-RT-PCR 

分 类 号：S436.43[农业科学—农业昆虫与害虫防治]