农杆菌介导将LycB基因转入优良玉米自交系的研究

Transfer of LycB from Lycium chinense Miller into Maize Excellent Inbred Line Mediated by Agrobacterium tumefaciens

以优良玉米自交天塔5的幼胚为材料,诱导出Ⅱ型愈伤组织,用带有质粒pCAMBIA2300-LmLycB-Bar根癌农杆菌C58侵染,对培养基和遗传转化因素进行优化;共培养3 d,延迟培养7 d,再转入含PPT的培养基上连续筛选3代,分化获得再生植株。经PCR和RT-PCR检测,确定获得13株阳性植株,HPLC检测转基因玉米叶片总类胡萝卜素含量为174.83μg/(g.FW),比野生型玉米提高了15.82%。

Use the excellent inbred - lines of maize Tianta 5 as the explant. The embryogenic type 11 callus was infected by Agrobacterium tumefaciens C58 containing plasmid pCAMBIA2300-LmLycB-Bar. We identified a set of factors to achieve an overall high efficiency of maize transformation and callus induction. After 3 days of co-cultivation and 7 days of resting, the immature embryos and callus were continuously selected on the medium containing phos- phinothricin（PPT） for 3 generations, and then plants were regenerated. It was proved by PCR and RT-PCR analysis that the target LycB gene had been integrated into the genome of regenerated plants. Ultimately, achieved and trans- genic maize leave showed 15.82% increase in total carotenoids and 54.38% increase in β-carotene.