Abstract: The paper aims to confirm the pathogens of grape crown gall in southern mountain area of Jinan, and provide a scientific basis for disease control. The pathogens were isolated and identified from crown gall of ‘Cabernet Sauvignon’grape with morphological and molecular methods. The results showed that twelve isolates were purified from the grape crown galls and were characterized as Agrobacterium tumefaciens strains according to the colony morphology on the selective MW medium. On the basis of physiological and biochemical characteristics, and the sequence analysis of 16S rDNA, the twelve strains were confirm to be Agrobacterium sp. The number of strains of each biotype was: biotype Ⅰ, 5, biotype Ⅱ, 4, and biotype Ⅲ, 3, Two highly virulence strains JC02 and JC05 were determined by inoculating carrot disc, sunflower and grape seedlings. The plasmid biologic identification showed that the JC02 and JC05 strains contained Ti plasmids, which were Octopus type. The results indicate that Agrobacterium vitis is the main pathogen of rape crown gall in southern mountain area of Jinan.