Abstract: The resistance to the black shank of 15 flue-cured tobacco lines including 6614, 8504, 9601, 9717, 8902-42,CF209, CF964, CF986, RGH12, YH01, YH02, YH05, YNH09, Yunyan97 and Yunyan98 was identified from 2001 to 2006 intermittently .To confirm the reproducibility of the identification of resistance to tobacco black shank,the artificial inoculation was used. The incubation and preservation of the black shank, and the preparation of inoculation medium were provided in this identification method. The time and quantity of black shank disease inoculation were also advised in this paper.The result showed that the resistance indexes of the lines were all below 25.The analysis of SSR showed that it was non-remarkable difference between the lines and the resistant CK,K326 and Gexin3.And the resistance was consistent among the years which showed that the way of the identification of resistance was reliable.