Abstract: The root, leaf and hypocotyl of aseptic seedlings derived from Astragalus membranaceus seed were used as explants, and the growth-regulating hormones 6-BA,2,4-D,NAA were set at different concentrations, the effects of different explants on callus induction rate, in vitro plantlet increment and rooting were studied, in order to provide reference for callus induction, plant differentiation and proliferation of different tissue explants of Astragalus membranaceus.The results showed that root, hypocotyl and leaf were the best explants for callus induction, of which, hypocotyl was the best explants, with the highest callus induction rate of 97.92%,at 2,4-D 1.0 mg· L^-1+6-BA 0.5 mg·L^-1+NAA 0.5 mg·L^-1+sucrose 3.0%,the best medium for plantlet proliferation at 1/2MS+6-BA 1.0 mg·L^-1+KT 0.5 mg·L^-1+sucrose 3.0%,the highest average differentiation rate of regenerated plantlets at 92.67%,and the optimum medium at 1/2MS+NAA 0.5 mg·L^-1+sugar 3.0%.The system of plant regeneration and multiplication of Astragalus mongholicus was established in this experiment,which provided a powerful technical reference for its germplasm resources preservation,rapid seedling breeding and development,which has important popularizing value in the production practice of Astragalus mongholicus.