Abstract: Taking the kiwifruit leaf spot disease as test materials,the pathogens were separated and purified by tissue culture method and verified to be the pathogen through an artificial infection test.Pathogens were identified by morphological and molecular biological methods,the biological characteristic of different conditions pathogen were studied,in order to provide a reference for the investigation of the disease occurrence and the formulation of comprehensive preventive measures.The results showed that Lasiodiplodia theobromae was the pathogen of kiwifruit leaf spot disease.The mycelia could grow at 10-35℃,and 28℃ was the optimum temperature,optimum pH was 5,lighting conditions were continuous light,the best carbon source for mycelial rowth was sluble starch,the best nitrogen source was yast extract.