猕猴桃溃疡病是猕猴桃生产上的主要病害,为建立该病的快速诊断技术,本实验通过RAPD分析获得一条1 300 bp左右的致病菌的特异片段,对该片段进行克隆测序,在测序的基础上设计并合成一对特异引物F7/R7,优化特异引物扩增条件,并验证引物的特异性和灵敏性。利用该特异引物对包括猕猴桃溃疡病菌在内的14个菌株基因组DNA进行PCR扩增表明,只有猕猴桃溃疡病菌能扩增出1条约为950 bp的特异条带,其他菌株及对照均未扩增出特异条带。对采自果园的染病枝干组织和接种致病菌的枝干组织的检测表明,该特异引物能特异性地检测到猕猴桃溃疡病菌的存在,其在组织中的检测灵敏度为100 fg/μL。因此,利用设计合成的特异引物F7/R7,参考优化的体系和程序,结合简单的试剂盒法提取猕猴桃溃疡病菌或植物组织DNA,可以在短时间内完成对该病原菌的分子检测。 Kiwifruit and canker disease is the main disease in the production of kiwifruit. In order to establish a rapid diagnostic technique for this disease, this experiment obtained a specific fragment of pathogenic bacteria with a length of about 1 300 bp by RAPD analysis. The fragment was cloned and sequenced, On the design and synthesis of a pair of specific primers F7 / R7, optimize the specific primer amplification conditions, and verify the specificity and sensitivity of the primer. PCR amplification of genomic DNA of 14 strains including Actinidia ulmoides showed that only one specific band with a length of 950 bp was amplified from Actinidia ulmoides and no other strains and controls were amplified Specific bands. The detection of diseased stem tissues and pathogen-inoculated branch tissues collected from orchards showed that this specific primer could specifically detect the presence of Actinidia kawachii and its detection sensitivity was 100 fg / μL in tissues. Therefore, the molecular detection of this pathogen can be completed in a short period of time by utilizing the specific primers F7 / R7 designed and synthesized, and referring to the optimized system and procedure, and using simple kit method to extract kiwifruit canopies or plant tissue DNA.