Abstract: Chilli veinal mottle virus (ChiVMV) is one of the main virus species in tobacco-growing areas of Southwest China, and its harm to tobacco has risen rapidly in recent years. In this study, the virus strain, disease phenotype and varieties resistance were identified by molecular biological detection and friction inoculation on tobacco seedlings. Results showed that the ChiVMV isolated from Liangshan and Baoshan tobacco were both severe strains causing yellow round spot and necrosis on leaves. The typical symtoms of ChiVMV-infected leaves appeared many round bright spots on leaves, with manifestation of chlorosis and yellowing. Moreover, the leaves shrank with droopy and folded leaf margin, especially for the lower leaves. Flue-cured tobacco varieties including K326, NC89, G28, Yunyan85, Yunyan87, Yunyan105, Yunyan116, Honghuadajinyuan and Zhongyan100, were susceptibility to ChiVMV, while N-genotype tobacco Samsun NN, CV91, and va-genotype tobacco VAM showed no obvious resistance to ChiVMV. Meitan sun-cured tobacco variety X45 exhibited moderate resistance to ChiVMV, with the virus concentration in its inoculated and systemic leaves significantly lower than that in susceptible varieties of N. benthamina and K326, thereby is an important resource for disease resistance breeding.