The Study On Genetic Transformation Of Anti-stresses Genes In Poplar

SnRK2 relatively small protein family which only plant have is subfamily of SNF1-related protein kinase,that is activated by osmotic stress and activate expression of downstream genes to improve stress resistence of plants.In this study,based on Genes of PtSnRK-1 and PtSnRK-2 that is cloned from Poplar trichocarpa and constructed into overexpression vectors,PtSnRK-1 and PtSnRK-2 are transferred into Populus×euramericana cv.'Nanlin895' to obtain new poplar that can stand stress tolerance.The mian conclusion of this experiment as following:1.Through Hygromycin sensitivity of screening test on differentiation of leaf and rooting of adventitious buds,and Cefradine sensitivity test on plants,determine that the optimal concentration for selection on differentiation of the leaves is Hygromycin 3mg/L,and the optimal concentration for screening of rooting is Hygromycin 1 mg/L,and the concentration of Cefradine which resitrain the growth of Agrobacterium tumefaciens EHA105 is 200mg/L,another Agrobacterium tumefaciens LBA4404 is 100mg/L.2.Genes of PtSnRK-1 and PtSnRK-2are transferred into Populus×euramericana cv.'Nanlin895' by Agrobacterium tumefaciens EHA105 and LBA4404,and got suitable condition for transformation.The suitable conditions of Agrobacterium tumefaciens EHA105 are as follow:pre-culture time is 2～3days,concentration of agrobecterium OD600 is 0.5～0.6,infection time is 15 min,con-culture time is 4 days.The suitable conditions of Agrobacterium tumefaciens LBA4404 are as follow:pre-culture time is2～3days,concentration of agrobecterium OD600 is 0.6～0.8,infection time is 20 min,con-culture time is 5 days.3.Transgenic poplars were selected to survived on the culture medium with antibiotics,and got got 9 positive plants transformed gene of PtSnRK-1 and 8 plants that transformed gene of PtSnRK-2 which tested by PCR anaylysis.Because of little time, advanced molecular detection and physiological analysis didn't make.