Abstract: The paper aims to test effects of medium with different hormone combinations on explants callus culture, buds proliferation and rooting culture of rare and endangered plants Ammopiptanthus with aseptic cotyledons and stem segments as explants materials, and screen the optimum medium formula for each culture stage. The results showed that cotyledon was the best explants to induct callus of Ammopiptanthus. The callus induction rate reached its maximum 81.8% at 6-BA 0.5 mg/L, 2,4-D 0.5 mg/L. The clustered buds proliferation rate reached the maximum of 90.2% at 6-BA 2.0 mg/L, NAA 0.5 mg/L. The phenomenon of multiple clustered buds, robust stem and vigorous growth was observed, indicating that higher concentrations of cell division 6-BA could significantly promote cluster bud proliferation. The rooting rate reached the maximum of 70.4% in 1/2MS basic medium added with NAA 1.0 mg/L, the rooting time was the earliest, with obvious main root, which was stout and hairy. The key point in psammophytes tissue culture research will be adjusting inorganic salt concentration in the medium properly to improve the rooting rate.