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本研究以优良玉米自交系交51为材料,通过农杆菌介导的茎尖转化将海藻糖-6-磷酸合成酶基因TPS1和葡萄花青素调控基因VlmybA2转入玉米中。确立了最佳的除草剂Basta筛选浓度,研究了不同真空渗透压、农杆菌液浓度、侵染时间等因素在转化过程中对玉米植株存活率的影响。结果表明80 mg/L除草剂Basta为最佳筛选浓度,真空渗透压为60 kPa、农杆菌液浓度OD600为0.8、侵染时间为8 min时是最佳转化条件,经过GUS组织化学染色和PCR检测,证明TPS1基因和VlmybA2基因已经整合到玉米基因组中。
In this study, the elite maize inbred lines to pay 51 as materials, through Agrobacterium-mediated shoot tip transformation trehalose-6-phosphate synthase gene TPS1 and grape anthocyanin regulation gene VlmybA2 transferred to maize. The best herbicide Basta screening concentration was established. The effects of different vacuum osmotic pressure, Agrobacterium concentration and infection time on the survival rate of corn plants were studied. The results showed that 80 mg / L herbicide Basta was the optimal screening concentration, the vacuum osmotic pressure was 60 kPa, the Agrobacterium concentration OD600 was 0.8, and the infection time was 8 min. After GUS histochemical staining and PCR TPS1 and VlmybA2 genes have been integrated into the maize genome.