Group F1 and F2 were constructed through hybridization and selfing by using Ti245 as the resistant parents and K326 as the susceptible parent. The inheritance of resistance to TMV from tobacco variety Ti245 was analyzed, and then SSR markers linked to the resistant gene were identified. The results showed the resistance of Ti245 to TMV was controlled by one pair of recessive genes. The region of the resistance of Ti245 to TMV was mapped on LG9; The distance between the *Tp217200 and the objective gene was the nearest, and the genetic distance was 3.3cM from the region of resistance to TMV.