Abstract: In order to obtain more antagonistic fungi for prevention and control of tobacco black shank, 20 fungal strains were isolated from tobacco rhizosphere soil, among which ZP2-1, ZP2-3, ZP2-4, GZ3, and GZ7 had antagonistic effects on Phytophthora nicotianae. Molecular and morphological analysis showed that these strains were identified as Trichoderma reesei ZP2-1, Trichoderma koningiopsis ZP2-3, Aspergillus niger ZP2-4, Aspergillus fumigatus GZ3 and Hypocre rufa GZ7. ZP2-4 has a strong ability to secrete cellulase and protease and the ability of sporulation in solid fermentation was higher than that of the other 4 strains (p<0.01). The cellulase activities of the solid fermentation products of ZP2-1, GZ3 and ZP2-4 were fairly high and significantly higher than that of ZP2-3 and GZ7 (p<0.01). ZP2-1, GZ3 and ZP2-4 with high spore producing ability and cellulase activity in solid fermentation were selected for field trial. The results showed that the control efficiency against tobacco black shank of ZP2-1, GZ3 and ZP2-4 were 59.50%, 61.82% and 85.00%, respectively.