不同转化方法培育无抗性选择标记转基因水稻效率的比较

Comparison on Efficiency of Generating Selectable Marker-free Transgenic Rice by Different Transformation Methods

为研究不同的转化方法培育无抗性选择标记转基因水稻的效果,以4个籼粳稻品种为受体材料,比较了用农杆菌介导的双菌双载体和双T-DNA单载体两种共转化技术的共转化频率及随后获得的无抗性选择标记基因转化子的效率。结果表明,在双菌双载体介导的转化中,4个水稻品种的平均共转化频率为10.1%,其中55.6%的共转化植株自交后代中可筛选出无抗性选择标记的转基因水稻植株。在单载体双T-DNA介导的共转化中,平均共转化频率为45.0%,从其中60.0%的共转化植株自交后代中可获得无抗性选择标记的转基因植株,即双T-DNA单载体法的去选择标记的效率(27.0%)要较双菌双载体法(5.6%)高。

The efficiencies of generating selectable marker-free （SMF） transgenic rice through two transformation methods were compared using four indica and japonica varieties as materials. One is by using a single twin T-DNA binary vector in one Agrobacteriurn strain. The twin T-DNA binary vector pYH592 is composed of two separate T-DNA regions, one carrying the antisense Wx gene while the other the HPT gene. The other one, known as two-vector system, is by using two separate binary vectors in two separate Agrobacteriurn cultures. The results indicated that the average co-transformation frequencies of the antisense Wx gene and the HPT gene were 10.1% and 45.0% for the four rice varieties, the SMF transgenic plants could be found in the offsprings of 55. 6% and 60. 0% co-transformants, and the effectivenesses of generating marker-free transformants were 5.6% and 27.0%, respectively, in the two-vector system and the twin T-DNA binary vector system.