番茄细菌性溃疡病菌的实时荧光PCR检测

Rapid detection of Tomato bacterial canker (Clavibacter michiganensis subsp. michiganensis) with real-time fluorescent PCR

由C lavibacter m ichiganensissubsp.m ichiganensis(Cmm)引起的番茄细菌性溃疡病是一种严重危害番茄生产的种传细菌性病害。根据ITS序列多态性设计引物及TaqM an探针进行实时荧光PCR检测的结果表明,这组引物-探针能检测出所有供试的Cmm菌,对照菌均未检测到荧光信号。用接种但未显示症状的番茄苗叶片及人工处理的带菌种子提取的核酸作为模板,均能检测到病菌,其检测灵敏度比常规PCR高约100倍。实验中不需病原菌的分离培养及PCR的后续处理。该方法快速、简便、安全、准确,适用于出入境检验检疫及种子、种苗健康检测领域。

Clavibacter michiganensis subsp, michiganensis is the most relevant tomato bacterial pathogen. The specific primers sets and TaqMan probe were designed based on the region of ITS to detect Cmm with real-time fluorescent PCR （RTF PCR）. The RTF PCR was approximately 100 times more sensitive and more specific than normal PCR. Strong fluorescent signal could be collected during the reaction that the DNA extracted directly from the latent infected seed and seedling were used as template. This method provides a specific, sensitive, rapid detection of Cmm for the seed and the seedling industry.