转基因水稻中重组植酸酶的表达

Expression of Recombinant Phytase in Transgenic Rice

为通过在转基因植株中表达植酸酶降解植酸来提高水稻中无机磷利用率,构建了由玉米泛素基因Ubi启动子控制的植酸酶基因植物表达载体,并以来源于水稻未成熟胚的愈伤组织作为转化受体,经农杆菌介导法将植酸酶基因导入水稻中,共获得15个独立的转基因株系。对转基因水稻总DNA的PCR和Southern杂交分析证明目的基因已整合入转基因水稻植株基因组中,并能稳定遗传。对部分转基因水稻未成熟种子总RNA进行RT-PCR分析,表明导入的植酸酶基因能够在转基因水稻种子中正常表达。无机磷含量分析表明含目的基因的转基因水稻种子及其后代叶片中的无机磷含量较未转化植株均有了明显的提高。

For improving inorganic phosphorus use efficiency by expression of phytase in transgenic rice, a plant expression vector, containing the recombinant phytase gene driven by the maize ubiquitin （Ubi） promoter, was constructed, and was introduced into rice via Agrobacterium-mediated transformation, by using the rice immature embryo-derived callus as the explants. A total of 15 independent transgenic lines were regenerated, and the results of PCR and Southern blot analyses showed that the target gene had been integrated into the genome of transgenic rice. RT-PCR analysis of the total RNAs extracted from the immature seeds of several transgenic lines showed that the introduced phytase gene could be normally expressed. The result of measurement of the inorganic phosphorus level implied that the inorganic phosphorus content was significantly higher in the mature seeds of several primary transformants and the leaves of their progeny than that of the untransformed control.