Abstract: ObjectiveIn this paper, the optimal medium of in vitro rapid propagation was selected in order to estab-lish rapid culture system for Sapindus mukorossi Gaertn seedlings . MethodTaking S. mukorossi selective stems with axillary buds as explants,the concentration of HgCl2, action time of HgCl2 and alcohol in explants sterilization, the con-tents of 6-BA, NAA and sucrose in adventitious bud induction, the dosages of 6-BA and KT in shoot proliferation, and inorganic salt levels in MS, dosages of 6-BA and 2,4-D in seedling rooting was optimized by orthogonal experiment design L9(33). ResultThe optimum stem sterilization method for S. mukorossi was as follows:immersing the stem in 75%alcohol for 1 min, and sterilized with 0.2% HgCl2 for 7 min. After this treatment, survival rate of explants was 83.33%. MS+2.0 mg/L 6-BA+0.1 mg/L NAA+20.0 g/L sucrose was the most appropriate culture medium for axillary bud induction, and the induction rate was 96.67%. MS+0.7 mg/L 6-BA+0.1 mg/L KT was the best culture medium for axillary bud proliferation, and the proliferation multiple was 4.13 times after 30 days. 1/3MS+0.5 mg/L 6-BA+0.7 mg/L 2,4-D was the best culture medium for rooting,and the average rooting rate was 41.50%. ConclusionThe in vitro culture system of S. mukorossi es-tablished preliminarily is as follows: immersing the stem explants in 75% alcohol for 1 min, sterilizing them with 0.2%HgCl2 7 min, using medium of MS+2.0 mg/L 6-BA+0.1 mg/L NAA+20.0 g/L sucrose to induce axillary bud germination, putting them in MS+0.7 mg/L 6-BA+0.1 mg/L KT medium for multiplication culture, and using 1/3MS+0.5 mg/L 6-BA+0.7 mg/L 2,4-D medium for rooting.