Abstract: The bud and basal part of the pseudostem of Kaempferia rotunda L.were used as materials.The effects of mediums with different hormone concentrations on bud proliferation, callus induction, proliferation and differentiation were studied by tissue culture, in order to provide reference for the establishment of a direct and indirect rapid propagation system for K.rotunda L..The results showed that the optimum medium for rapid proliferation was MS+6-BA 2.0 mg·L^-1+NAA 0.1 mg·L^-1 which had large proliferation coefficient and strong buds.In the indirect plant propagation system, the best medium for callus induction was MS+6-BA 2.0 mg·L^-1+2,4-D 1.0 mg·L^-1.Callus induction rate was up to 78.43% after induced from basal part of pseudostem for 40 days.MS+6-BA 1.0 mg·L^-1+2,4-D 2.0 mg·L^-1 produced the highest callus proliferation rate.The most suitable callus differentiation was MS+6-BA 4.0 mg·L^-1+NAA 0.6 mg·L^-1,with the differentiation rate being the highest（66.67%）.The tissue cultured plants were transferred onto humus after hardening-seedling, and the survival rate was higher than 90%.Direct and indirect rapid propagation system of seedling for tissue culture of Kaempferia rotunda L.was established, which provide the technical support for factory seedling, genetic breeding and specific medicinal ingredient bioreactors.