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以杂交育种中广泛使用的优良玉米自交系 34 0、4112为材料 ,用带有质粒pGBIL0 4(Pactin -Bt-Tnos)的根癌农杆菌LBA44 0 4转化其幼胚及其初始愈伤组织 ,共培养 3天后 ,在含PPT的培养基上连续筛选培养 3代 ,然后分化获得再生植株。PCR检测证明目的基因已整合到再生植株的基因组中。实验结果表明幼胚预培养后形成的新鲜的初始愈伤组织是比较适宜的转化受体 ,结果还发现将共培养温度降到 2 2℃可以提高农杆菌介导的玉米遗传转化的筛选频率。
The immature embryos and their initial callus were transformed with Agrobacterium tumefaciens LBA44 0 4 with the plasmid pGBIL0 4 (Pactin -Bt-Tnos) using the maize inbred line 34 0,4112 widely used in crossbreeding. After three days of co-culture, the cells were continuously screened and cultured on the medium containing PPT for three generations and then differentiated to obtain regenerated plants. PCR test proved that the target gene has been integrated into the genome of the regenerated plant. The results showed that fresh initial callus formed after pre-culture of embryos was a suitable transformation acceptor. The results showed that reducing the co-culture temperature to 22 ℃ could improve the frequency of Agrobacterium-mediated maize genetic transformation.