Abstract: The tissue culture system of high-yielding Sambucus williamsii was established using young stem segment of cutting seedlings as the material, aimed to speed up the breeding of improved varieties. The results showed that the proper method for surface sterilization was treating the stem segment of cutting seedlings with 75% alcohol and 0.1% HgCl2 for 30 s and 15 min, and the survival rate could reach 80%. The medium suitable for bud germination was MS+ NAA 0.05 mg/L, with the induction rate of 98.7%. The optimal culture medium for multiplication was MS+ 6-BA 1.0 mg/L+ TDZ 0.01 mg/L+ NAA 0.1 mg/L, with multiplication coefficient of 3.82. The rooting medium was 1/2MS+ IBA 0.1 mg/L, with the best growth and the rooting rate of 100%. When transplanted, a combination of perlite and humus in the ratio of 3:5 was adopted as the medium, and the survival rate could reach over 95%.