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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.
Agrobacterium tumefaciens with activating marker and Basta screening marker plasmid pSKI015 was transformed by high yielding and good yielding rapeseed variety 1244W6 and the mutant library of rapeseed activating marker was successfully constructed.After Basta screening, 000 transgenic lines (T1 generation) .Several transgenic lines (T2 generation) were cultured in nutrient solution containing no hormones or plant hormones with different concentrations. As a result, 63 hormone- The hypocotyls and short hypocotyl mutants were 15 and 13, respectively. The TAIL-PCR technique was used to clone the adjacent sequence of the genomic DNA of the W1016 hormone-reactive mutant T-DNA insertion site.