兰花褐斑病菌实时荧光PCR检测  

Detection of Acidovorax avenae subsp. cattleyae by real-time fluorescent PCR

作　　者：丁翠珍[1] 赵文军[1] 寸东义[2] 陈红运[3] 朱水芳[1] 

机构地区：[1]中国检验检疫科学研究院动植物检疫研究所,北京100029 [2]云南出入境检验检疫局,昆明650228 [3]厦门出入境检验检疫局,厦门361012

出　　处：《植物病理学报》2010年第3期235-241,共7页Acta Phytopathologica Sinica

基　　金：质检公益性行业科研专项(200810632);"十一五"国家科技支撑计划(2006BAK10B06)

摘　　要：兰花褐斑病菌(Acidovorax avenaesubsp.cattleyae)是兰花上一种重要进境检疫性有害生物,可通过植株和种子远距离传播,其传染性强,对兰花危害性大。根据核糖体ITS序列设计了TaqMan-MGB探针并建立了实时荧光PCR检测方法。该方法能够特异性检测兰花褐斑病菌,所有供试的目标菌株检测结果均为阳性,而其它28个对照菌株(含同属内其它种及avenae种下其它亚种)均为阴性。利用该方法从发病兰花植株总DNA中检测到该病菌。本方法灵敏度高,检测极限达9×10-9μg DNA,操作方便快速,结果可靠,适合于口岸兰花的进出境检疫及兰花种苗健康质量控制。Acidovorax avenae subsp.cattleyae（AACa）,the pathogen causing bacterial brown spot of orchid,is a quarantine organism in China.However,with conventional methods it is difficult to distinguish this pathogen quickly and reliably from other closely related pathogen in the genus of Acidovorax.In this study,a TaqMan-MGB probe was designed based upon the ITS sequence of Acidovorax and a real time PCR method was developed.AACa can be distinguished from other 28 bacteria strains including other species of genus Acidovorax and subspecies in avenae with this method.The absolute sensitivity threshold was approximately 9×10-9 μg bacterial genomic DNA.By this method AACa could be detected easily from infected orchid plant directly without bacteria culture.This real time PCR method could assist in the implementation of quarantine measures for prevention and control of the disease caused by AACa.

关 键 词：兰花褐斑病菌 TAQMAN-MGB探针 实时荧光PCR 检测 

分 类 号：S432[农业科学—植物病理学]