Abstract: In order to study the tobacco brown spot disease resistance inheritance, and genetic mapping, F₂ and BC₁ populations from a cross between Tobacco Brown Spot resistant variety Jingyehuang and susceptible variety NC82 were developed. Values of disease index under artificial inoculations showed that the resistant trait of Jing-yehuang was fitted additive-dominance model. Meanwhile, a further molecular study was carried out using SSR markers based on a former genetic mapping, and a SSR marker which was tightly linked to the tobacco brown spot resistant gene located on M linkage groups with distance of 4 cM was obtained. This marker provides useful information for molecular marker assistant selection of brown spot resistance in breeding program of tobacco.