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In the present study, a plant regeneration system was established by using young leaf explants from aseptic seedlings of Cassia mimosoides. The results showed that when the explants were inoculated on the 1/2MS medium contained 2.0 mg/L 6-BA and 0.5 mg/L NAA, high rate of callus induction and good growth status of adventitious bud, and the highest number of buds were obtained. Further studies conclusively suggested that IBA could not promote root formation effectively, but the root regeneration could be induced by NAA. However, although the rooting of regeneration buds could be induced when adventitious buds were placed into 1/2MS medium supplemented with 0.5 mg/L NAA, the rate of rooting was reduced, and the roots became shorter. Therefore, the best medium for adventitious bud regeneration using young leaf as explants in C. mimosoides was 1/2MS medium contained 2.0 mg/L 6-BA and 0.5 mg/L NAA, and the best rooting medium was 1/2MS.