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为了诱导重金属镉超富集植物油菜毛状根形成并获得较高的转化率,利用正交试验(L8(27))优化油菜毛状根诱导的最佳条件,正交实验结果表明:利用发根农杆菌ATCC11325对预培养48 h的油菜子叶进行转化,转化时间10 min,加入0.3 mg/L的2,4-D激素时,油菜毛状根的诱导率可达到73%。PCR扩增显示农杆菌Ri质粒中的rol C基因已经插入并整合到油菜核基因组中,成功建立了油菜毛状根体系。
In order to induce the formation of hairy roots and to obtain higher conversion rate of rapeseed (Cd), the optimal conditions for hairy root induction were optimized by orthogonal experiment (L8 (27)). The results of orthogonal experiment Agrobacterium tumefaciens ATCC11325 transformed the rape cotyledons pre-cultured for 48 h and the transformation time was 10 min. The induction rate of hairy roots of rapeseed reached 73% when 0.3 mg / L 2,4-D was added. PCR amplification showed that the rol C gene in Agrobacterium Ri plasmid had been inserted into the nuclear genome of rape and successfully established the hairy root system of rapeseed.