The antagonistic bacteria strains By33 and By88 were isolated from top soils of tobacco planting fields. The results showed that the invitro inhibition of sterile fermented broths of By33 and By88 to TMV was 81.81% and 83.29% respectively, and 73.01% and 66.98% respectively inoculating TMV 24h after smearing fermented broths. The antagonistic proteins obtained by ammonium sulfate deposition had the control of 85.38% and 87.47% to TMV, with absorption apexes in 274 nm. The study also showed that the combination of antagonistic proteins and TMV was inreversible, and could restrain the copy of TMV in tobacco.