Establishment Of Regeneration System And The Preliminary Study On Genetic Transformation System Of Purple Dioscorea Alata L.

Dioseorea alata Z.,belong to Dioscoreaceae Dioscorea vine,with the high yield potential.lt was an imporant food crop, a new chioce of biomass energy for its hight starch content, and the pueple Dioscorea alata L. is a good source of pigment.In this study,select the purple Dioscorea alata L.from the Dioscorea resource garden of Hainan University,college of Agriculture as the material to study the tissue culture techniques of purple Dioscorea alata L.,initially established the rapid propagation and regeneration system;study the Agrobacterium mediated purple Dioscorea alata L. genetic transformation system.The research results are as follows:1Rounding1-2internodes near the shoot tips of the pueple Dioscorea alata L.seeding,and the remaining internode in tap water rinse30min,75%alcohol disinfection10s,0.1%HgCl2disinfection8min, the survival rate of the internoderate can be as high as100%.2Purple Dioscorea alata L.seeding stem segments cultured on hormone-free MS medium supplemented with10days germination to budding,30days out a stem section.The nodal stem of subcluture can propagation rapidly in the medium of MS+80mg/L AdSO4,grow4internodes about60days.3The internodes cultured from the MS+80mg/L AdSO4can be induced adventitous buds at least7in the medium of MS+6-BA1.0mg/L+NAA0.1mg/L+80mg/L AdSO4about30days,but the adventitous buds grow very slowly in this medium.Transferred the adventitious buds to the medium of MS+80mg/L AdSO4, it can propagation4internodes about50days. In the medium of MS or MS+NAA0.1mg/L,the adventitous buds can take root well.4The leaves, stems and bulbil of purple Dioscorea alata L.seedlings only can induce the loose callus, the nodal stem of plantlet can induce the callus which can redifferentiation the adventitous buds.The nodal stems induce the rat of compact callus as49.17%in the medium of MS+6-BA1.0mg/L+NAA0.2mg/L.Compact callus differentiatious the rate of adventitous buds as95.00%in the medium of MS+6-BA0.3mg/L+NAA0.06mg/L. 5The callus of purple Dioscorea alata L. after genetic transformation of Agrobacterium OD value of0.5, the infection time30min,co-cultured4d, the survival rate of callus as42.50%in the selective medium of MS+6-BA0.3mg/L+NAA0.06mg/L+40mg/L hygromycin+300mg/Lcefotaxime.