油菜突变体库构建与激素反应基因克隆分析

Construction of the Mutant Library of Brassica Napus and Cloning of Phytohormone Response Genes

以优质高产的油菜品种1244W6为材料,采用浸花蕾方法,将含有激活标记和Basta筛选标记质粒pSKI015的农杆菌进行转化,成功构建了油菜激活标记突变体库.通过Basta筛选,结果共获得约30 000个转基因株系(T1代).将部分转基因株系种子(T2代)培养在不含任何激素、或含有不同浓度植物激素的营养液中,结果筛选出激素反应突变体63个,长下胚轴和短下胚轴突变体分别为15和13个.采用TAIL-PCR技术,克隆了W1016激素反应突变体T-DNA插入位点基因组旁邻序列.

An activation tagging mutant library of Brassica napus 1244w6 was constructed in the inflorescence dipping method, and pSKI015 plasmid with Basta screening marker was transformed in the Agrobacterial tumefaciens containing method. 30 000 independent T1 transformants were obtained through basra screening. 63 phytohormone response mutants were screened by sowing seeds of T2 transformants in nutrient solution con- taining different content of phytohorraone. 15 long hypocotyl and 13 short hypocotyl mutants were screened by sowing seeds of 72 transformants in nutrient solution without phytohormone. The T-DNA flanking sequence of phytohormone response mutant W1016 was cloned by using TAIL-PCR method.