The effects of different kinds of reagents and disinfection time,hormone combination and hormone concentration gradient on the tissue culture of Siraitia grosvenorii were studied by using tender stems as experimental materials.The best disinfection way for Siraitia grosvenorii is 0.1% HgCl₂ solution disinfecting 6 min,efficiency of sterilization can reach 85%.The tender stems were cultured on the medium of 0.5 mg/L 6-BA + 0.1 mg/L NAA,and the seedlings with large shoots were obtained,and the stem segments of the seedlings were removed as explants for further cultivation.It was found that the 0.4 mg/L IBA + 1 mg/L 6-BA was the best subculture medium,and the multiplication coefficient was up to 6.3.The best culture medium of shoots development was 0.05 mg/L 6-BA + 0.1mg/L GA₃,the average elongation of 35 d was 5.1 cm.The best rooting medium was 1/2MS + 0.5 mg/L IBA + 0.5mg/L NAA,and the number of rooting was 3-8.Rooting rate was up to 86%.