根癌农杆菌介导的糖苷转移酶基因转化烟草的条件研究

Research of Conditions on Tobacco Transformation Mediated by Agrobacterium Tumefaciens Expressing Glycosyltransferase Gene

采用根癌农杆菌介导的烟草叶盘转化法,研究了外源糖苷转移酶基因导入烟草、建立糖苷转移酶基因的转基因烟草体系过程中,影响根癌农杆菌转化频率的各种因素.结果表明,植物材料需预培养3～5 d,共培养约3～5 d较合适;农杆菌需稀释至最适浊度O.D.600约为 0.5;抑制农杆菌生长的氨苄青霉素终浓度为600 mg·L-1.

To transfer a glycosyltransferase gene,whose expression is induced by methyl jasmonate, into tobacco leaf discs of W38 with the help of Ti-plasmid of Agrobacterium tumefaciens, the transformation conditions were optimized. It was demonstrated that 3~5 d pre-culture and co-culture of tobacco leaf discs were needed for the procedure of this gene transformation. In addition, the optimal concentrations of Agrobacterium tumefaciens solution and ampeicin for inhibiting the growth of Agrobacterium tumefaciens  should be approximately 0-5 of O.D.600 and the 600 mg·L -1 for the processes of affection and selection, respectively.