梨轮纹病和炭疽病病原菌PCR检测

摘要

为建立梨轮纹病原菌和炭疽病病原菌的分了检测方法,根据GenBank上炭疽病原菌和轮纹病原菌不同种的ITS序列差异设计2对引物TJ1/T J2和LW1/LW2,对2种病原菌菌株进行扩增,比较了常规PCR和巢式PCR的检测灵敏度,并对果园中接种梨轮纹病原菌的梨果实进行检测.结果表明,建立的PCR体系可分别从炭疽病原菌和轮纹病原菌菌株扩增出317 bp和325 bp的特异性条带,对其他相似或相近的病原真菌则无扩增条带.巢式PCR技术的灵敏度比常规PCR提高了约105倍,且可以在接种较低浓度(1ml10个)轮纹病原菌孢子液7d后的梨果实组织中检测到病原菌.说明使用巢式PCR技术,可以准确、灵敏地监测梨轮纹病原菌在果园的种群消长动态.%Based on the differences in internal transcribed space (ITS) sequences of Colletotrichum genus and Bot-ryosphaeriu genus, two pairs of species-specific primers TJ1/TJ2 and LW1/LW2 were synthesized. Two specific bands of 317 bp and 325 bp were amplified from C. gloensporioides and B. berengeriana, respectively, while other strains displayed no band. The detection sensitivity of nested-PCR was 105-fold higher than that of regular PCR. DNA extracted from pears sprayed with low concentraion (1 ml 10 U) of conidial suspension of B. Berengeriana for 7 d could be detected by nested-PCR. It indicated that nested-PCR could stably and quickly monitor the population growth dynamic of pathogenic bacteria in orchard.