Establishment Of Regeneration System In Vitro And Preliminary Exploration Of Agrobactirium-mediated Transformation System Of P.Simonii Carr.×P.Deltoides Cv.’shanhaiguanensis’

In recently years,’GM gene deletion’system had becoming a prospectively popular tool for solving the biological safety of transgenic plants. The objective of this study was to establish a highly efficiency in vitro regeneration system and Agrobactirium-mediated transformation system and adventitious buds’rooting system using leaf and petiole as explants. The results will support the later study of Populus simonii’s genetic breeding. In order to develop new technology for Populus simonii gene preservation and enhancing its resistance to disease and pests, leaves of hybrid clone off. simonii Carr.×P. deltoides cv.’Shanhaiguanensis’were used as explants to study culture and leaf regeneration system in vitro and to explore the effect of different prescriptions of hormone on induction, differentiation of adventitious bud and rooting. The main results that we had gained were as follows:1. The optimum medium for adventitious buds regeneration of P. simonii Carr.×P. deltoides cv.’Shanhaiguanensis’was MS+0.1mg/L6-BA+0.08mg/L NAA+0.005mg/L TDZ, with the maximum adventitious bud frequency (90%), number of shoots (6.8), shoot length (2.14cm). The optimum medium for rooting of shoots was1/2MS+0.1mg/L IBA, with a rooting efficiency of90%. Thus the obtained plantlets were successfully hardened and transplanted to greenhouse.2. We established a regeneration system using petiole as explant and analyzed the influence of differentiation and adventitious bud’rooting in different hormone combinations. The results showed that:the optimum medium for adventitious buds regeneration of the Populus simonii Carr.×P. deltoides cv.’Shanhaiguanensis’was MS+0.1mg/L6-BA+0.02mg/L NAA, with the maximum adventitious bud frequency (90%), number of shoots (6), shoot length (2.25cm).3. Put foliages’face up using leaves as explant led to the best regeneration efficiency. The best leaves and petioles for adventitious buds regeneration all came from the upper stems.4.20mg/L kanamycin could inhibit the bud regeneration of "P. simonii Carr.×P. deltoides cv.’Shanhaiguanensis’" when using leaf as explants,10mg/L kanamycin could inhibit the bud regeneration of "P. simonii Carr. XP. deltoides cv.’Shanhaiguanensis"’ when using petiole as explants,20mg/L kanamycin could inhibit the root regeneration of P. simonii Carr. XP. deltoides cv.’Shanhaiguanensis’from the shoots.5.The results of bacteriostasises effect of Cef research showed that:When the concentration of Cef reached to200mg/L, this concentration could restrain the growth of agrobacterium and had no obvious side-effects on adventitious bud regenerations and rooting of adventitious buds of the P. simonii Carr.×P. deltoides cv.’Shanhaiguanensis’. So, we had chosen the200mg/L Cef as the best antibacterial concentration.