Study On The Establishment Of Regeneration System Of Salix Linearistipularis

Salix linearistipularisy belongs to the Salix,Salicaceae,is adaptively grown in the saline-alkali area and can be characterized by the high salt resistant and the improvement of high pH soil.However,as the plant is distributed in the limited area and is not grown well in the drought and rainless area,it is not easy to propagate them in large amounts naturally.In addition,for the more in-depth and efficient study on the salt resistant genes of S.linearistipularis,it is necessary to establish the effective plant regeneration and genetic transformation system of S.linearistipularis.In this study,the regeneration system of S.linearistipularis plants from the internodes was established,and the results were as follows:1.The propagation system of aseptic S.linearistipularis plants was successfully developed.The terminal and lateral buds were cultivated in different cultural conditions for the propagation.The best response was obtained from the cultivation of terminal buds on the 0.1 mg/L IBA adding MS basal medium.On the other hand,the lateral buds were also grown well on the MS basal medium containing little amounts of 6-BA,IBA and TDZ.2.The effects of different kinds of plant hormones on the callus induction were analyzed.The combinational effects of NAA+6-BA,2,4-D+KT and NAA+6-BA+TDZ were mainly considered in this study.In the combination of NAA and 6-BA,the MS media supplemented with 0.1 mg/L NAA and 0.9 or 1 mg/L 6-BA resulted in 100%callus induction and proliferation rates,and the induced callus was relatively in good condition.The general induction rates were higher in the combination of 2,4-D and KT than that of NAA+6-BA.Especially,the media containing 0.5 mg/L 2,4-D+0.8 mg/L KT and 1 mg/L 2,4-D+2 mg/L KT showed best induction results.In addition,the addition of different concentrations of TDZ to the combination of NAA and 6-BA resulted in 100%induction rates,indicating the promoting effect of TDZ on the callus induction.3.Zebularine known as a kind of DNA methylation inhibitor has a positive effect on the callus induction,therefore,20 different kinds of media which recorded high induction rates were specifically selected and 1 mg/L zebularine was added for the bud induction.The results showed that the adventitious buds were successfully induced in 6 kinds of media containing 1 mg/L zebularine.Especially,under the conditions of MS+0.2 mg/L NAA+0.2 mg/L 6-BA and MS+0.5 mg/L 2,4-D+0.8 mg/L KT the induction states were relatively good.The adventitious buds induced were successfully elongated on the condition of 0.1 mg/L NAA+0.1 mg/L 6-BA,and the root was finally induced on the 0.1 mg/L IBA-containing MS medium.Through the overall results,the preliminary study on the establishment of regeneration system of S.linearistipularis was successfully completed,and it can provide the fundamental data for the further research on the efficient genetic transformation system of this plant.