农杆菌介导的獐茅NHX1基因导入水稻辽粳9号体系的建立及优化  

Establishment and Optimization of agrobacterium-mediated Transformation System in Rice Liaojing 9 Via Transforming NHX1

作　　者：丰明[1] 王旭达[1] 王鹤[1] 李怀梅[1] 张高华[1] 

机构地区：[1]辽宁省农业科学院大连生物技术研究所,辽宁大连116029

出　　处：《沈阳农业大学学报》2012年第3期326-330,共5页Journal of Shenyang Agricultural University

基　　金：辽宁省博士科研启动基金项目(20101054)

摘　　要：离子平衡调节是植物抵御盐分伤害的一种非常重要的机制。将已分离出的獐茅Na+/H+逆向转运蛋白基因AlNHX1构建到植物表达载体pCAMBIA1301中,通过农杆菌介导转化法将AlNHX1基因转入到辽粳9号水稻中,并对转化体系进行优化。结果表明:水稻愈伤组织诱导培养基中2,4-D的最适浓度为2mg.L-1,最佳侵染时间为90s;抗生素敏感性试验结果表明:筛选培养基中适宜的潮霉素选择压为50mg.L-1。在较优转化体系下,辽粳9号水稻的转化频率和再生频率分别达到43.9%和16.1%。Ion homeostasis plays an important role in plant salt torlerance.In this study,the vacuolar Na＋/H＋ antiporter gene AlNHX1 from a graminaceous halophyte Aeluropus littoralis（Gouan）Parl was constructed into plant expressed vector pCAMBIA1301.The AlNHX1 was transferred into Liaojing 9 via Agrobacterium-madiated method,and the transformation system was optimized.The results showed that the optimum 2,4-D concentration in the callus inducing medium was 2mg.L-1,with the optimum infection time 90s.The experiment on sensitivity of cotyledon to hygromycin B showed the suitable hygromycin B concentration for selecting transgenic tissue was 50mg.L-1.We established an efficient transformation system on Liaojing 9 with a relatively high transformation frequency of 43.9% and the regeneration frequency of 16.1%.

关 键 词：NA+/H+逆向转运蛋白 农杆菌介导转化 水稻 

分 类 号：S511[农业科学—作物学]