长岭发垫刃线虫(Trichotylenchus changlingensis)双重PCR检测方法研究  

Development of duplex PCR assay for detection of Trichotylenchus changlingensis

作　　者：郭宁[1] 石洁[1] 马红霞[1] 刘树森 张海剑[1] 赵宝广 GUO Ning;SHI Jie;MA Hong-xia;LIU Shu-sen;ZHANG Hai-jian;ZHAO Bao-guang(Plant Protection Institute of Hebei Academy of Agricultural and Forestry Science;Key Laboratory of IPM on Crops in Northern Region of North China,Ministry of Agricultur;IPM Centre of Hebei Province,Baoding 071000,Chin)

机构地区：[1]河北省农林科学院植物保护研究所农业部华北北部作物有害生物综合治理重点实验室河北省农业有害生物综合防治工程技术研究中心,保定071000

出　　处：《植物病理学报》2018年第4期572-576,共5页Acta Phytopathologica Sinica

基　　金：现代农业产业技术体系(CARS-02)

摘　　要：长岭发垫刃线虫（Trichotylenchus changlingensis）是一种迁移性植物外寄生线虫,该线虫于2011年首次在我国吉林省长岭县发现,并根据其形态特征及最新分类系统将其归为发垫刃属[1,2]。该线虫能引起玉米叶片黄化,植株矮化,茎基部开裂,茎节缩短等症状,该病发生率普遍在21%～67%,给玉米生产造成了严重影响[3]。In order to simply,rapidly and accurately identify Trichotylenchus changlingensis,the ITS region of 8 different populations of T.changlingensis was amplified by PCR using a pair of universal primer F194/5368r of nematode.Subsequently,a pair of specific primer,TC-F1/TC-R1,was designed according to the ITS sequence of T.changlingensis and used to establish a duplex PCR system with the combining primer pair D2A/D3B which was a universal primer to detect plant parasitic nematodes and used as internal reference in this study.The results of this study showed that a fragment of 1057 bp was amplified from 8 T.changlingensis populations using the primer pair F194/5368r,and a specific band of 520 bp was amplified by TC-F1/TC-R1 which displayed a high specificity.Under the duplex PCR system,the anticipated PCR products obtained from T.changlingensis were 520 bp and 780 bp,respectively,and the detection sensitivity was 0.125 ng·μL-1 of DNA.This system showed the advantages of high sensitivity,strong stability,time-saving and simple operation,and is suitable for the rapid detection of T.changlingensis.

关 键 词：寄生线虫 PCR检测方法 长岭县 玉米生产 新分类系统 形态特征 叶片黄化 植株矮化 

分 类 号：S432.45[农业科学—植物病理学]