To identify the pathogenic bacteria of kiwifruit bacterial canker, the infected kiwifruit branches and barks from Anhui (Yuexi County) Province,Shaan’xi (Huxian County) Province and Chongqing (Qianjiang County) Municipality were collected as the experiment materials. Plate streaking, gradient dilution and beef extract peptone agar (BPA) media cultivation were used for bacteria isolation, and tobacco hypersensitive responses detection, kiwifruit pathogenicity detection and 16S-23S rDNA-ITS sequential analysis were carried out as follows for bacteria identification. From the bacterial canker infected kiwifruit tissues, 10 pathogenic strains were isolated, and all isolated strains have pathogenicity and hypersensitivity to tested kiwifruit and tobacco leaves. However, different isolated strains have different pathogenicity for kiwifruit. 10 purified pathogenic strains belonged to the same pathogen through 16S-23S rDNA-ITS and Blast sequential analysis. The amplified 280 bp sequences were the same as the previously reported 16S-23S rDNA-ITS of Pseudomonas syringae pv.actinidae. It is concluded that the isolates from the canker infected kiwifruit branches and barks were Pseudomonas syringae pv.actinidae.