[Objective] The aim was to protect wild resource of Atractylodes lances and to provide technical guidance for industrialization of Atractylodes lances. [Method] The test-tube seedlings were produced from seeds of Atractylodis Lances. At the same time, the effects of 6-BA, NAA and KT on the proliferation of aseptic seedling was studied through the orthogonal test. Effect of NAA concentration on the rooting of aseptic seedling was studied through nine types of light substrate groups, which consist of peat soil, fine sand and cotton seed hull, and their effects on Atractylodes lances growth were studied on with orthogonal design. [Result] The culture medium suitable for the proliferation of the Atractylodes lances tube seedling was MS＋6-BA 1.0 mg/L＋NAA 0.4 mg/L＋KT 0,4 mg/L. And the medium suitable for rooting was 1/2MS＋NAA 0.5 mg/L. The optimal ratio of soil, riversand and cotton seed hull was 3：3：2. [Conclusion] The research selected optimum culture medium, rooting medium and ratio of substrate, which provided technical guidance for laboratorial preservation of germplasm resource of Atractylodes lances and industrialization of its seedling production.
