Indirect somatic embryogenesis and plant regeneration of Hippophae rhamnoides sub. sinensis.

Using the shoot tips of seabuckthorn as explants,the callus,embryogenic callus and somatic embryos were induced on 1/4 MS basal medium and the effects of different plant growth regulator combinations on callus,embryogenic callus induction and somatic embryogenesis were studied.Results showed that the best combination was KT 1.0 mg/L + NAA 0.3 mg/L,on which the induction frequency of callus and embryogenic callus reached 90.10% and 77.67% after 20 and 30 days culturing in darkness,respectively.When the embryogenic callus were transferred to fresh induction medium and cultured under the light,the average number of somatic embryogenesis was 15.48 after 20 days.In order to get the regeneration plantlet, the embryoids were transferred to the medium for somatic embryo development and plant regeneration.The effects of KT combinations and different concentrations of granulated sugaron plant regeneration were analyzed.The frequency of plant regeneration was 60.30% on the medium of WPM + KT 0.05 mg/L + granulated sugar 20 g/L + agar 6.5 g/L + CH 0.5 g/L.In this study,a complete system for plant regeneration via indirect somatic embryogenesis from shoot tips of seabuckthorn was achieved.