A Real-time PCR Assay with Cycling Fluorescent Probe for Detection of Carbendazim Resistance in Fusa

• 作 者: HouYi-ping;LuoQing-quan;ChenChang-jun;WangJian-xin;ZhouMing-guo;
• 机 构: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095
• 出 处: 中国植物病理学会化学防治专业委员会第七届中国植物病害化学防治学术研讨会
• 发表日期: 2010年期

　　The resistance of F.graminearum to benzimidazole fungicide carbendazim (MBC) is caused by point mutations in the β2-tubulin gene (FGSG_ 06611.3).The point mutation at coden 167 (TTT→TAT, F167Y), a dominant single mutation (more than 90%) resulted in moderate resistance.Based on this point mutation, two molecular techniques were developed to detect the MBC-resistant genotypes.A pair of primers Codon167F/Codon167R was designed to amplify a frag ment containing the mutative site in the first technique, and the two Cycling probes labeled different fluorescent reporter were developed in the second technique to detect whether the site mutate.In the study, the result of one hundred isolates collected from Tongzhou, Jiangsu in 2006-2007, detected by real-time PCR (Cycling probes) were confirmed by the conventional method.The realtime PCR with cycling probes is rapid, high-throughput, quantitative to survey the frequency of the MBCMR mutation (F167Y) genotpye ofF.graminearum.