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以玉米自交系天塔五母、7922的芽尖为受体,用农杆菌介导法将psy基因转入玉米中。以植株的转化率为指标,研究了真空处理方式,真空处理时间及共培养时间对转化率的影响。结果表明,当在真空条件下侵染20min,共培养3天时转化率最高。用200mg/L的草胺膦溶液筛选,获得抗性植株经PCR检测有16株表现阳性,其中天塔五母有12株,7922有4株。RT-PCR及高效液相色谱(HPLC)检测结果表明psy基因已经整合进玉米基因组并能正常转录,转基因玉米中总类胡萝卜素含量比野生型玉米提高了25%。
The psy gene was transferred into maize by agrobacterium-mediated method using the bud tips of 7922 as the top five parents of maize inbred lines. The conversion rate of plant was used as an index to study the effects of vacuum treatment, vacuum treatment time and co-culture time on the conversion rate. The results showed that the transformation rate was the highest when co-cultured for 3 days under the condition of 20 min inoculation under vacuum. Screening with 200mg / L sodium phosphinothricin solution, the resistant plants obtained by PCR detected 16 positive, of which there are 12 towers of the tower, 7922 have 4. The results of RT-PCR and HPLC showed that psy gene was integrated into maize genome and transcribed normally. The content of total carotenoids in transgenic maize increased by 25% compared with that of wild-type maize.