Abstract: In order to establish the technology system for the tissue culture and rapid propagation of Acacia podalyriifolia, the response surface method (RSM) was used to optimize the adventitious bud induction, proliferation and the rooting medium formula. Based on the previous single factor pre-test, the induction rate, proliferation coefficient and rooting rate were taken as the response values. The response surface methods of Central Composite Design (CCD) and Box-Behnken (BBD) test design were used to optimize the optimal hormone ratio for the adventitious bud induction, proliferation and rooting culture of Acacia podalyriifolia. The results showed that the optimum induction medium was MS+3.0 mg·L^-1 6-BA+0.35 mg·L^-1 IBA+0.2 mg·L^-1 GA₃, and the induction rate was 62.89%. The optimal proliferation medium was 1/2MS+0.66 mg/L 6-BA+0.45 mg·L^-1 IBA+0.11 mg·L^-1 GA₃, and the proliferation coefficient was 3.78. The optimal rooting medium was 1/2MS+0.64 mg·L^-1 IBA+0.14 mg·L^-1 NAA, and the rooting rate was 79.91%. At the same time, the actual value was found to be in line with the expectation according to the contour line map and response surface analysis of the interaction between the factors of response surface method. In conclusion, the prediction of the regression equation established by using the response surface method had a good fitting degree with the actual experiment, which could be used to optimize the medium conditions for tissue culture of Acacia podalyriifolia. The transplanting substrate was yellow subsoil+sand (the volume ratio of yellow subsoil and sand was 2∶1), and the transplanting survival rate could reach more than 90%.