应用分子生物学方法快速检测番茄溃疡病菌的研究

Study on Rapid Identification of Clavibacter michiganensis subsp. michiganensis by Molecular Biological Method

根据番茄溃疡病菌的tomA基因序列,设计并合成了特异性PCR检测引物,对番茄溃疡病菌和非番茄溃疡病菌的标准菌株进行了PCR扩增反应。结果,番茄溃疡病菌的PCR产物出现301bp的特异性扩增条带,而非番茄溃疡病菌均未出现扩增条带,证明这对引物具有番茄溃疡病菌鉴定特异性。将分离的番茄溃疡病菌做梯度稀释,测定该检测体系的敏感度。结果表明,此体系可检出106CFU/mL番茄溃疡病菌菌液提取的模板。

A rapid molecular biological identification method of Clavibacter michiganensis subsp, michiganensis is established by using one pair of primers unique to tomA gene segments. A specific DNA amplicon was present in the standard strains, but absent in the non - standard strains. It showed that this pairs of primers was specific for rapid identification of C. michiganensis subsp, michiganensis. The sensitivity of this method was confirmed by testing the serial dilutions of C. michiganensis subsp, michiganensis, which could be determined at the concentration of 10^6CFU/mL.