Single PCR reaction with two pairs of PCR primers designed on the markers respectively tightly linked with Cf-9 and Tm-1 genes in tomato, which be resistant to leaf mold and tomato mosaic virus. For the genes the PCR products were almost completely correspond to the amplified bands produced by single PCR primer. Among them, 560bp fragment linked with Cf-9 gene was amplified in both resistant and susceptible tomato lines. The amplified bands from susceptible and resistant lines were distinguishable after cleavage with the restriction enzyme TaqI. Genotype with Cf-9 gene could produce respectively 450bp, 330bp and 290bp bands. Susceptible genotypes could produce respectively 450bp and 290bp fragments. 750bp fragment linked with Tm-1 gene was amplified in resistant tomato line. The amplified bands could not be cleavaged with the restriction enzyme TaqI. The replicated stable results proved that two resistant genes could be identified simultaneously by using corresponding PCR primer under adaptable condition. This system compared with single primer PCR would be time saving, less labor and low cost. It could bevery useful for marker-assisted selection during early stage in tomato and efficiently speed up breeding procedure.