Abstract: Tobacco mosaic virus (TMV) is one of the major diseases on tobacco, spreading fast and popular early. Mechanical contact infection in farm work and transmission of virus residues in soils are the major culprits for the rampant outbreak of TMV. For application of bio-control bacteria in soil and instruments disinfection, the suppression in vitro of the five single bacteria cultures and the twain bacteria cultures on TMV was investigated on local lesion host of Nicotiana tabacum var. samsum NN challenged by mechanical inoculation of mixture of cultures with TMV. The results showed that both of the five single bacteria cultures and the twain bacteria cultures had good disinfection effect on TMV, all of 5-fold dilution cultures suppressed TMV from 94% to 99% in vitro compared to the mixture of NB with TMV. And the twain bacteria cultures of CZ and 2A2 had the best disinfection effect, with 74.90% of suppression in vitro by 50-fold dilution cultures. These results indicated that CZ and 2A2 has a potential to be an instruments and soils disinfector against TMV.