Abstract:Sporophytes of Alsophila spinulosa were obtained from spore propagation agamospory and adventitious bud produced from callus by tissue culture. The morphology of prothallus is plastic. In the tissue culture, the prothallus had three forms; normal long cordate prothallium, abnormal ligulate prothallium and cylindrical prothallium, and then they could produce normal plant. When BM was a improved MS, the spore germination, filamentous growth and transition from one to two-dimensional growth, prothallium development and sporophyte differentiation etc. were influenced by sugar concentration, inorganic salts concentration, composition and proportion of hormone.