利用同源转基因技术培育氮高效利用转基因水稻  

Breeding of high efficient nitrogen utilization transgenic rice through intragenesis technology

作　　者：王虹玲[1,2] 阚国仕[1] 李珊珊[2] 马小路[1] 贾士荣[2] 刘昱辉[2] 

机构地区：[1]沈阳农业大学生物科学技术学院,辽宁沈阳110161 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《浙江农业学报》2011年第5期862-869,共8页Acta Agriculturae Zhejiangensis

基　　金：转基因生物新品种培育科技重大专项(2008ZX08009003)

摘　　要：为了提高水稻对氮素的吸收利用率,培育不含任何标记基因、报告基因和载体骨架序列的转基因水稻,从硅藻中克隆了硝酸盐转运蛋白(Diatom Nitrate Transporter,NAT)基因,构建了3个植物表达载体:可使NAT在水稻中高效表达但无抗性筛选标记基因的植物表达载体pANW2300K-、带有负选择标记—大肠杆菌胞嘧啶脱氨酶CodA的正负筛选植物表达载体pCodA1301和植物表达载体pIpt121。明确Ipt基因可显著抑制转基因烟草的生长发育,但对水稻的影响较小;建立了以短暂选择和正负筛选为基础的水稻同源转基因技术;获得了18株有目标基因NAT,但不含任何筛选标记基因和载体的骨架序列的转基因水稻植株,转化率达27.7%;实现了同源转基因技术在水稻中的应用,这在国内尚属首例。In order to improve rice＇s nitrogen absorption rate, marker-free, reporter-free and backbone-free transgenic rice, （Nitrate Transporter, NAT） was cloned from diatom genome, and three plant expression vectors were constructed： marker-free plant expressing vector pANW2300K- which makes it possible for diatom nitrate transporter- NAT gene to be highly expressed in rice, positive-negative plant expressing vector pCodA1301 which carries a negative selectable marker-CodA and the plant expressing vector plptl21. Through these vectors we proved that lpt gene can inhibit the growth of tobacco, while having little effect on rice. We also established a transgenic system based on transient selection and positive-negative selection. Eighteen marker-free and backbone-free rice plants expressing the target gene NAT were identified to be transgenic, representing a transformation rate of 28%. It is the first time in China that intragenesis technology was fully used in rice.

关 键 词：水稻 同源转基因技术 短暂选择方法 正负筛选 NAT 

分 类 号：S511[农业科学—作物学]