摘要
番茄溃疡病是一种严重危害番茄生产的细菌性病害,许多国家将其列为检疫性病害。利用ITS通用引物扩增了番茄溃疡病菌(Clavibacter michiganensis subsp.michiganensis)的ITS序列,并进行克隆测序。根据序列比较结果设计了引物BT1和BT2,该引物特异性好,能专一扩增出268bp电泳条带,而马铃薯环腐病菌等不同亚种、不同属的细菌及健康的番茄材料均无扩增条带。从接种但未显症番茄苗叶片及人工模拟染菌种子上提取总DNA,以此为模板均能稳定地扩增出特异性目的条带。该方法直接对种子或植株进行检测,不需进行病原菌分离培养,快速简便,适用于出入境检验检疫及种苗健康检测领域。
Bacterial canker and wilt of tomato, caused by Clavibacter michiganensis subsp, michiganensis （Cram）, is one of the most serious diseases of glasshouse and field tomatoes. EPPO has listed Cmm as an A2 quarantine pest, and CPPC and IAPSC also consider it of quarantine significance. Seed is the main long- distance vector and seed trade has facilitated the worldwide distribution of the disease. A PCR protocol was developed to specifically detected the pathogen based on the ITS sequence. The DNA directly extracted from seeds and seedling without symptom is used as template of PCR reaction, the pathogen can be detected and identified from other subspecies of C. michiganensis . This method provides a rapid and reliable tool for routine detection and identification of Cmm in plant quarantine fields.
出处
《植物检疫》
2007年第2期75-77,共3页
Plant Quarantine
关键词
番茄溃疡病菌
ITS
PCR
检测
Bacterial canker and wilt of tomato, ITS, PCR, detection
