Abstract: Taking aseptic induced buds of Anoectochilus roxburghii stem as experimental material，callus and protocorm were induced form shoot tip and stem on germinal bud，respectively.Aseptic cultivate system Anoectochilus roxburghii of was established.And optimum medium for micropropagation was found.Results showed that optimum medium for bud germinate of Anoectochilus roxburghii stem was MS+BA 1.5 mgL+banana juice 100 gL+NAA 0.5 mgL.Optimum medium for protocorm induction of shoot tip was MS+BA 2.0 mgL+NAA 0.3 mgL+HyponcexⅠ3.0 gL.Optimum medium for subculture micropropagation of protocorm was 12MS+BA 2.0 mgL+NAA 0.3 mgL+banana juice 100 gL+AC 0.2%.Optimum medium for cluster buds induction from callus was 12MS+BA 2.0 mgL+NAA 0.2 mgL+banana juice 100 gL.Optimum medium for subculture micropropagation of cluster buds was 12MS+BA 3.0 mgL+NAA 0.5 mgL+KT 1.0 mgL+banana juice 100 gL+AC 0.2%.Optimum medium for rooting of plantlets was 12MS+IBA 1.0 mgL+NAA 0.3 mgL+AC 0.2%.