农杆菌介导的小麦遗传转化条件的研究  

Optimization of the Conditions for Wheat Genetic Transfermation Mediated by Agrobacterium Tumefaciens

作　　者：刘庆法[1] 唐克轩[1] 叶建明 郝峥嵘[1] 何艺园[1] 沈大棱[1] 

机构地区：[1]复旦大学遗传学研究所

出　　处：《复旦学报（自然科学版）》1998年第4期569-572,共4页Journal of Fudan University：Natural Science

摘　　要：利用来自小麦花药、幼胚愈伤组织和带有GUS基因的Ti质粒，对农杆菌转化小麦的条件进行了初步研究，证实适当浓度的乙酰丁香酮是诱发T－DNA转移的必需条件，其最低有效浓度为50μmol／L．在20～35℃，pH5．2～6．8条件下，均可有效诱导T－DNA的转移．农杆菌菌液密度以108ml－1细胞为宜，添加适当的表面活性剂有促进T－DNA转化的作用．对4种小麦材料的研究证明，T－DNA的转移效率常因受体材料基因型的不同而有很大差异；愈伤组织的活力与T－DNA转移的效率有密切关系．显微观察发现，胚状体细胞比非胚状体细胞群更容易发生转化．The conditions for wheat genetic transformation mediated by Agrobacterium tumefaciens were optimized using calli derived from anthers and immature embryos and a Ti plasmid containing the GUS gene. The presence of acetosyringon in co-culture medium was essential for the efficient transfer of T-DNA into callus cells,with the lowest effective concentration of 50 μmol/L. A high incidence of T-DNA transfer was closely associated to the physiological state of callus. The coculture temperature from 20℃ to 35℃ and the medium pH from 5. 2 to 6. 8 have the same T-DNA transfer efficiency. The inoculated bacterium cell intensity was optimized as 108 cells/ml. In addition, it was found that surfactant could enhance T-DNA transfer.

关 键 词：小麦 农杆菌介导 基因转化 遗传转化条件 

分 类 号：S511.032[农业科学—作物学] Q78[生物学—分子生物学]