提高小麦基因枪法转化频率的研究  

Studies of Improving the Frequency of Wheat Transformation by Biolistic Bombardment

作　　者：梁辉[1] 唐顺学[1] 张弛[1] 赵铁汉[1] 李良材[1] 欧阳俊闻[1] 田文忠[1] 王道文[1] 贾旭[1] 

机构地区：[1]中国科学院遗传研究所,北京100101

出　　处：《Acta Genetica Sinica》1999年第6期643-648,共6页

摘　　要：用基因枪法将带Bar－GUS双标记基因的质粒（pAHC25）转入普通春小麦品种中─60634的幼胚盾片，并获转基因植株。在轰击的经预培养3～4天的342块幼胚盾片再经筛选再生的植株中，经PCR和Southern分析表明，外源基因已稳定整合到小麦基因组中，转化率为0．88％。此外发现，幼胚培养基中用2mg／Ldicamba代替2mg／L2，4－D，可提高愈伤组织的再生能力。同时，为了保持小麦幼胚愈伤组织的分化能力，在诱导愈伤组织阶段5mg／Lbialaphos的筛选时间以不超过1个月为宜，及早转入3mg／Lbialaphos分化及壮苗培养基筛选转基因植株。In the present study immature embryos of spring wheat Zhong-60634 were bombarded with gold particles coated with pAHC25 containing Bar and GUS genes. A total of 17 bialaphos-tolerant plants were obtained from 342 immature embryos which were first cultured for 3-4 days and then bombarded and selected. Integration of the introduced genes into the genome of bosgenic wheat plants was shown by PCR and Southern analysis and transformation frequency was 0.88%. Furthermore, the present study showed that calli regeneration ability was improved while the embryo culture medium was supplemented with 2mg/L dicamba instead of 2mg/L 2,4-D. Meanwhile,in order to keep the calli differentiation ability calli should be transfered to differentiation meditum supplemented with 3mg / L bialaphos from calli selection medium with 5mg/L bialaphos within a month.

关 键 词：小麦 转化频率 DICAMBA 基因枪法 转基因工程 

分 类 号：S512.103[农业科学—作物学] Q789[生物学—分子生物学]