[ Objective] The pathogen of a new disease found in floating seedlings of tobacco was isolated and identified to provide the basis for the control of the disease. [ Method] The pathogenicity of the isolated strain was determined according to Koch's rules, and the pathogen was identified through Biolog system, 16S rDNA sequence analysis and physiological and biochemical methods. [ Result] Through Koch's test, the isolated 3 -3 strain was verified to be the pathogen causing floating seedling disease, which was consistent with the characteristics of Pectobacterium carotovo- rum subsp. Carotovorum through Biolog determination and the other physiological and biochemical methods. 16S rDNA sequence analysis showed that 3 -3 strain had the highest similarity with P. carotovorum subsp. Carotovorum strains Kun28213 (accession number GU936996), reaching 99. 9%. [Conclusion] Base on the identification results of several methods, the pathogen causing floating seedling disease was P. carotovorum subsp. Carotovorum, and the disease was first reported in China. According to the English name of the disease, the disease was called as tobacco blackleg disease.