Using the young segments of Menyanthes trifoliata L.as explants transferred to MS medium with different concentration hormone(6-BA/NAA) can introduce the calli,buds and root growth.The study found that explants cultured on MS medium with 1.0 mg/L 6-BA and 0.1 mg/L NAA after 10 days can be observed in light green calli tissue.Calli transferred to MS added 4.0 mg/L 6-BA and 0.3 mg/L NAA for 2 weeks produce buds.Only the calli with buds could induce roots to form the complete regeneration plants.The most suitable root inducing medium is MS medium added 0.3 mg/L 6-BA and 1.0 mg/L NAA.Plant micropropagation technology successfully established regeneration system of M.trifoliata L.in this experiment,and it provides a new way for the large-scale production of M.trifoliata L.strains.