农杆菌介导水稻幼胚转化获转基因植株

AGROBACTRIUM-MEDIATE TRANSFORMATION OF RICE IMMATURE EMBRYOS AND RENGERATION OF TRANGENIC PLANTS

本研究以根癌农杆菌LBA4404(pTOK233)为供试菌株,以水稻幼胚为供试材料,对影响农杆菌介导基因的转化效率的因素进行了研究,确立了农杆菌介导的水稻高效转化体系。采用所确立的转化体系转化了Radon、中国91、024283个水稻品种(系)幼胚,结果表明,gusA基因的瞬时表达率均在70%左右,最高为76%;gusA基因稳定表达率均在20%以上,平均为23%。转基因植株的GUS活性检测及Southern杂交分析表明,外源基因已整合于水稻基因组中,进行了有效地表达,且能稳定地遗传给后代,外源基因在后代遗传符合孟德尔遗传规律。

We used Agrobactriumstrain LBA4404 harboring PTOK233 which contains gusA gene and hpt gene to infect rice immature embryos After the factors influencing transformation carefully had been investigated,a highly efficient Agrobacterium-mediated gene transfer system was developed Using this protocol,immature embryos of Radon,Zhongguo 91 and 02428 were infected and transformed The results showed that as high as 76% of transient transformation frequency and 23% of stable transformation frequency were obtained Southern blot analysis and GUS activity assay of transgenic plants showed the gusA gene had been integrated into the rice genome,and had expressed effectivly The results of genetic analysis showed that the transgeneswere segregated in a Mendelian fashion in the T 1 generation