Abstract: Two resistant × susceptible hybrid combinations were made between the line resistant to black shank (Beinhart1000-1) and two susceptible materials, SNN (Samsun NN) and XHJ (Xiaohuangjin1025). All the three parents and the two crosses were performed black shank artificial inoculation and resistance evaluation at mature stage. The genetic characteristics of Beinhart1000-1 resistance were studied using the major gene plus poly gene model. The results showed that: The black shank resistance genetic model of the cross Beinhart1000-1 × SNN was identified to fit model E2 and the heritability of major genes and poly-genes were estimated to be 99.14% and 0.48%, respectively. For cross Beinhart1000-1 × XHJ1025, it fitted B1 model very well, and the heritability of major genes was estimated to be 99.52%. Because of the higher heritability of the major genes, the selection of black shank resistant plants could be done in early generations. These results could provide information on MAS for improvement of resistance to black shank disease in tobacco.