Through the ‘Real gifted one’ of Hippophae rhamnoides varieties with 1~2 years old and dormant buds stems (axillary bud of unerupted) as explants，different culture，different types of basic culture medium and different concentration of hormone combination on Hippophae rhamnoides tissue culture influence in the process of systematic were studied.The results showed that the stem segment culture medium for differentiation was 1/3MS+6-BA 0.5 mg/L+IAA 0.3 mg/L；and the induction rate could reach more than 85%；the best of proliferation and induction of callus culture medium was 1/3MS+6-BA 0.75 mg/L+IAA 0.5 mg/L；callus induction rate in 90% above，and proliferation coefficient reached 4~6；the optimum rooting medium was 1/4MS+NAA 0.3 mg/L+IBA 0.5 mg/L；average rooting rate above 85%.After inoculation dark culture was added into the culture medium 7 d，6.5 g/L agar，1.5 g/L activated carbon，more training method could effectively prevent the browning.