Objective To establish the system of inducement, subculture, and plantlets regeneration of callus from Salvia miltiorrhiza. Methods Compared with the effect of different explants, basic media, and plant growth regulator on the inducement, subculture of callus, and the differentiation of buds. Results MS+6-BA 2 mg/L+NAA 0.2—2 mg/L was propitious to the inducement of callus and the ratio of induced callus, for both the leafstalk and lamina, was 100%, but quantity of induced callus was more of The ratio of buds differentiation was 55.6% on the medium of MS+6-BA 2 mg/L, and it could grown into buds directly. B5+6-BA 1 mg/L+2, 4-D 1 mg/L was the better subculture medium of callus. ConclusionSo for the callus inducement, the better explant and medium are leafstalk and MS+6-BA 2 mg/L+NAA 0.2—2 mg/L, respectively; For the buds differentiation, the better explant and medium are lamina and MS+6-BA 2 mg/L, respectively; For the subculture of callus, B5+6-BA 1 mg/L+2, 4-D 1 mg/L is better.