梨轮纹病和炭疽病病原菌PCR检测

为建立梨轮纹病原菌和炭疽病病原菌的分子检测方法,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/TJ2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(1 ml 10个)轮纹病原菌孢子液7 d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。

Detection of Botryosphaeria berengeriana and Colletotrichum gloeosporioides by PCR technology

Based on the differences in internal transcribed space(ITS) sequences of Colletotrichum genus and Botryosphaeria 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.gloeosporioides 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.