The induction and differentiation were studied on the callus of cotyledon of Cynanchum paniculatum and rooting and transplanting of test-tube plantlets as well as establishment of asexual system of Cynanchum paniculatum were successfully carried out in the experiment.The results suggested:MS+6-BA 0.3 mg/L+NAA 0.2 mg/L+2,4-D 0.4 mg/L was the optimum medium for callus induction and subculture of cotyledon.The ideal medium for callus differentiation was MS+6-BA 0.8 mg/L+NAA 0.2 mg/L.The medium of MS+6-BA 0.6 mg/L+NAA 0.2 mg/L was optimal for differentiation and subculture of adventitious buds.And the cider was used as ideal matrix for plantlets culture with vigorous and variant characters.Based on the result of this experiment,induction and regeneration system of Cynanchum paniculatum could be set up.