Tobacco is widely planted across the world especially in China, which means that a large amount of tobacco waste needs to be treated. This study investigated the biogas fermentation of tobacco stalks co-digested with different biomass feedstocks and the inactivation of Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV) by anaerobic digestion. Results showed that the maximum methane yield of tobacco stalks at 35 degrees C was 0.163 m(3) CH4.kg VS-1, which was from the co-digestion of tobacco stalks, wheat stalks and pig manure. The largest VS removal rate of tobacco stalks was 59.10%. Proven by indicator paper stripe, half-leaf lesion and RT-PCR, CMV could be inactivated by mesophilic and thermophilic anaerobic digestion, whereas TMV could be only inactivated by thermophilic anaerobic digestion over 20 days. These results suggested that using tobacco stalks as feedstock for anaerobic digestion and applying the digested residue and slurry to Solanaceae crop land are feasible. (C) 2015 Published by Elsevier Ltd.