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通过对已有的油菜农杆菌遗传转化体系的改进 ,建立了一种利用农杆菌感染油菜子叶柄的高效遗传转化方法 ,经过农杆菌感染的子叶柄在含有 6 BA 2 .0mg/L ,IBA 0 .15mg/L ,AgNO33 .5mg/L的MS培养基上由卡那霉素抗性愈伤组织分化出抗性幼苗的频率达到 12 .2 8%。利用已经构建的含有油菜Napin启动子加上反义ACP脱饱和酶第一外显子、内含子的基因片段的植物双元表达载体 pNACP ,将该反义 ACP基因片段转入油菜“沪油 15” ;PCR和GUS染色结果表明抗性植株中有 60 %的植株含有导入的目的基因。这为油菜籽脂肪酸遗传改良提供了一种新的方法
Through the improvement of the existing Agrobacterium tumefaciens genetic transformation system, a highly efficient genetic transformation method using Agrobacterium tumefaciens to infect petiole of rapeseed was established. After the Agrobacterium tumefaciens infected cotyledonary petiole containing 6 BA 2 .0 mg / L, IBA 0 The frequency of resistant seedlings differentiated from kanamycin-resistant callus on MS medium containing 15 mg / L and AgNO33.5 mg / L reached 12.28%. The constructed plant binary expression vector pNACP containing the napin promoter of rapeseed plus the first exon and intron of antisense ACP desaturase was used to transfer the antisense ACP gene fragment into oilseed rape 15 "; PCR and GUS staining results showed that 60% of the plants in the resistant plants contain the introduced gene of interest. This provides a new method for genetic improvement of rapeseed fatty acids