The effects of different antibiotics (kanamycin, hygromycin B and carbenidillin) on induction and differentiation of callus (Saussurea involucrate), was discussed in this study and Saussurea involucrate leaves were used as explants. A doses strategy of selection and bacterium inhibition during agrobacter mediated genetic transformation was established. The result showed that the kanamycin of 40 mg/L was significantly influenced on callus growth, when it was 50 mg/L, the growth of callus was inhibited completely; the hygromycin of 8 mg/L was the best concentration for inhibiting the callus growth, the callus were geatly damaged and it’s characters were small, browning and dying with the concentration of hygromycin up to 20 mg/L. Meanwhile, the low concentration (0.5-2.0 mg/L) of hygromycin can enhance the rate of callus differentiation. Carbenidillin is an antibacterial agent of Agrobacterium, carbenidillin had apparently influnced on growth of callus at different concentrations, and when the concentration of carbenidillin was greater than 400 mg/L, the growth of callus was effected observably. This paper may provided a theoretical for S. involucrate genetic transformation system. The effects of different antibiotics (kanamycin, hygromycin B and carbenidillin) on induction and differentiation of callus (Saussurea involucrate), was discussed in this study and Saussurea involucrate leaves were used as explants. A doses strategy of selection and bacterium inhibition during agrobacter mediated genetic transformation was established. The result showed that the kanamycin of 40 mg / L was significantly influenced on callus growth, when it was 50 mg / L, the growth of callus was inhibited completely; the hygromycin of 8 mg / L was the best concentration for inhibiting the callus growth, the callus were geatly damaged and it’s characters were small, browning and dying with the concentration of hygromycin up to 20 mg / L. Meanwhile, the low concentration (0.5-2.0 mg / L) of hygromycin can enhance the rate of callus differentiation. Carbenidillin is an antibacterial agent of Agrobacterium, carbenidillin had apparently influnced on growth of callus at different concentrations, and wh en the concentration of carbenidillin was greater than 400 mg / L, the growth of callus was observed as observably. This paper may provide a theoretical for S. involucrate genetic transformation system.