西瓜细菌性果斑病病菌活细胞PMA-PCR检测方法的建立  

Establishment of detection method for viable cells of Acidovorax avenae subsp. citrulli with PMA-PCR

作　　者：赵璟源 胡佳续[1] 张莹[1] 刘鹏[1] 唐芳[1] 廖芳[1] 

机构地区：[1]天津出入境检验检疫局,天津300461

出　　处：《中国植保导刊》2015年第9期12-15,共4页China Plant Protection

基　　金：天津出入境检验检疫局科研课题(TK071-2013;Tk073-2013)

摘　　要：利用叠氮溴化丙锭与聚合酶链式反应结合(PMA-PCR),建立了西瓜细菌性果斑病病菌(Acidovorax avenae subsp.citrulli)活细胞检测方法。实验结果表明,当PMA浓度为0.5 mg/m L时,对活细胞DNA的PCR扩增无影响,最低检测浓度为1×104cfu/m L。当样品中死亡细胞浓度低于107 cfu/m L时,PMA可抑制死亡细胞DNA的PCR扩增;当死亡细胞浓度为1×107cfu/m L时,PMA失去效果。该方法适用于低浓度西瓜细菌性果斑病病菌活细胞的检测。Detection method for viable cells of Acidovorax cwenae subsp, citrulli （Aac） was established with propidiummonoazide （PMA） and PCR. The results showed that PCR amplification of DNA from viable cells was not inhibited by PMA with concentration of 0.5 mg/mL, and minimum detectable concentration of Aac was 104 cfu/mL. PCR amplification of DNA from dead cells was inhibited by PMA with suspensions of dead Aac containing less than 107 cfu/mL. PMA treatment was ineffective with suspensions of dead Aac containing at 1 ×10^7 cfu/mL. The method was applicable for detection of low concentration of living cells of Aac.

关 键 词：西瓜细菌性果斑病病菌 叠氮溴化丙锭 聚合酶链式反应 活细胞检测 

分 类 号：S436.42[农业科学—农业昆虫与害虫防治]