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 avenaesubsp.cattleyae is an important quarantine pest on orchids that can be transmitted by plants and seeds over long distances. It is highly contagious and harmful to orchids. The TaqMan-MGB probe was designed based on the ribosomal ITS sequence and a real-time fluorescence PCR detection method was established. The method can detect the brown spot pathogen specifically, all the tested target strains tested positive, while the other 28 control strains (including other species in the same gene and other subsp. Avenae species) were negative. The method was used to detect this pathogen from the total DNA of diseased orchid plants. The method has high sensitivity, detection limit of 9 × 10-9μg DNA, convenient and fast operation, reliable results and is suitable for the entry and exit quarantine of port orchid and orchid seedling health quality control.