利用PCR技术专化性检测瓜类细菌性果斑病菌  

Specific Detection of Acidovorax avenae subsp.citrulli by PCR

作　　者：田艳丽[1] 胥婧[1] 赵玉强[1] 李新辉 胡白石[1] 刘凤权[1] 

机构地区：[1]南京农业大学植物保护学院植物病理学系,农业部作物病虫害监测与防控重点开放实验室,江苏南京210095 [2]新疆维吾尔自治区福海县农业技术推广站,新疆福海836400

出　　处：《江苏农业学报》2010年第3期512-516,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点基础研究发展计划项目(2009CB119200);农业部“948”项目(2008-Z30);全国科技支疆计划项目(200840102-03)

摘　　要：将hrpB2基因作为检测靶标,根据hrpB2基因设计了1对特异性引物HB2F2/HB2R2,用此特异引物从瓜类细菌性果斑病菌菌株中扩增出290 bp的特异性片段,而其余参试菌株和甜瓜组织的PCR反应结果为阴性,灵敏度试验证明可以检测到103CFU/ml靶标菌体。对市售的17个品种的甜瓜种子进行PCR检测,其中4个品种检测出携带有瓜类细菌性果斑病菌。将hrp基因作为靶标,为快速检测瓜类细菌性果斑病菌提供了新的方法。A subspecies-specific polymerase chain reaction assay was developed for detecting Acidovorax avenae subsp.citrulli（Aac） in melon seed.Novel subspecies-specific primers were designed based on the sequences of the hrpB2 gene from Aac genomic DNA and tested for specific DNA amplification by PCR.The primers amplified target band from all Aac strains and yielded no amplicons with other tested bacteria.The PCR protocol was also tested for their ability to detect Aac in naturally diseased plant tissues,including leaves,fruits and seeds.It successfully detected the target bacteria as few as 10^3 cfu/ml.In addition,4 of 17 melon seedlots collected from the markets were found positive of carrying Aac by this assay.The PCR protocol provides a rapid and reliable tool for routine detection and identification of Aac strain.

关 键 词：瓜类细菌性果斑病菌 hrpB2基因 特异性引物 PCR检测 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治]