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以小麦绵阳19和洛阳8716幼胚愈伤组织为材料,利用gus基因的瞬时表达,探讨了农杆菌菌株和侵染条件对转化效率的影响。结果发现,对于农杆菌菌株EHA105,小麦绵阳19和洛阳8716幼胚愈伤组织侵染的最佳条件均为OD6000.8,侵染30min。对于农杆菌菌株LBA4404,绵阳19和洛阳8716幼胚愈伤组织侵染的最佳条件分别为OD6001.0,侵染30min和OD6000.8,侵染60min。利用建立的农杆菌转化体系对小麦高分子量麦谷蛋白亚基1Bx14基因进行了转化,经潮霉素抗性筛选、PCR和PCR-Southern杂交检测,平均转化率为0.61%。
The effects of Agrobacterium tumefaciens strain and infection conditions on the transformation efficiency were studied by using the immature embryos callus of Mianyang 19 and Luoyang 8716 as materials. The results showed that the optimal conditions for Agrobacterium tumefaciens strain EHA105, wheat Mianyang 19 and Luoyang 8716 immature embryo callus were OD6000.8, infection 30min. The optimal conditions for Agrobacterium strain LBA4404, Mianyang 19 and Luoyang 8716 immature embryo callus were OD6001.0, infection 30min and OD6000.8, infection 60min. The high molecular weight glutenin subunit 1Bx14 gene of wheat was transformed by Agrobacterium tumefaciens transformation system. The hygromycin resistance screening, PCR and PCR-Southern hybridization showed that the average transformation rate was 0.61%.