PCR-based specific detection of Clavibacter michiganensis subsp.michiganensis

• 作 者: XUE Qingyun ZHANG Xiaomei WU Xinhua Neil C.Gudmestad GUO Jianhua
• 机 构: Department of Plant Pathology,College of Plant Protection,Nanjing Agricultural University;Engineerin
• 出 处: 第五届中国植物细菌病害、第七届中国植物病害生物防治暨国际细菌学及植物病害生物防治学术研讨会
• 发表日期: 2010年10期
• 关键词: Clavibater michiganensis subsp.michiganensis PCR detection ITS primers CmmF1/Cmm

　　Internal transcribed spacer(ITS)regions of 16 strains of 16 species representing 20 strains of Clavibater michiganensis subsp.michiganensis(Cmm)and 38 other related bacterial strains were amplified by using eubacteria universal primers U1/U2.After these fragments were analyzed,a pair of primers,CmmF1/CmmR2 specific to Cl.michiganensis subsp.michiganensis was designed and used in this experiment.By this pair of PCR primers,an expected 393 bp DNA fragment was amplified from 20 Cmm strains,while no PCR product was obtained from other tested 38 bacterial strains.Primers CmmF1/CmmR2 was sensitive enough to detect 10 pg templates DNA or 103 CFU/mL Cmm,and to catch one infected tomato seed from one hundred seeds.Up to the present,this new PCR-based specific system with these new primers is the most sensitive one for detection of Cl.michiganensis subsp.michiganensis from bacterial suspension and infected tomato seeds.