【Objective】In order to protect wild germplasm resources and fulfill seedling demands for artificial cultivation, the present experiment was conducted to develop a regeneration system for Atractylodes japnica using young stem. 【Method】The young stem of A. japnica was used as explants to investigate the effects of different culture media and hormone combinations on callus induction and differentiation, rooting of adventitious buds, multiplication of test-tube plantlets and transplantation. 【Result】Among different culture media supplemented with different hormone combinations， viz.， MS， 1/2MS， 8114 and B5，1/2MS+6-BA 0.8 mg/L mg/L+NAA 0.5 mg/L+2，4-D 1.5 mg/L and 1/2MS+6-BA 0.8 mg/L+NAA 0.5 mg/L+2，4-D 2.0 mg/L were optimum for callus induction, and 1/3MS and 1/3MS+ IAA 0.2 mg/L were ideal for callus differentiation. Of the 1/2MS， 1/4MS, N6 and B5, LS media supplemented with NAA （0.6 mg/L） and IAA （0.6 mg/L） and 1/4MS with IAA （0.3 mg/L） was optimum for rooting of adventitious buds and subculture multiplication of test-tube plantlets. The rooting rate was recorded as 92% and the average multiplication coefficient was 2.7. The test-tube plantlets were easy to be transplanted and survive， and showed 98.6% survival rate. 【Conclusion】The regeneration system for Atractylodes japnica was established and the transplanted plantlets grew vigorously, and flowered and fruited next year with all characteristics of wild plant.