农杆菌介导的水稻双载体共转化法中部分影响因素的研究  

A study on Several Factors Influencing Co-transformation of Rice with Two Agrobacterium Binary Vectors

作　　者：陆美芳[1] 刘巧泉[1] 于恒秀[1] 顾铭洪[1] 

机构地区：[1]扬州大学江苏省植物功能基因组学重点实验室

出　　处：《生物技术通报》2005年第5期55-62,共8页Biotechnology Bulletin

基　　金：国家"863"高技术研究计划(2004AA212092);国家自然科学基金(30470992和30300226资助项目)

摘　　要：以T-DNA区分别只含有潮霉素选择标记基因(HPT)和GUS报告基因的双元载体pCAMBIA1300和pCAMBIA0301用于农杆菌介导的水稻共转化试验。根据经农杆菌浸染并共培养3d后水稻愈伤组织中的GUS瞬间表达情况及其稳定共转化率,测定了不同农杆菌菌株搭配及其不同浓度配比对共转化效率的影响。结果表明,在两种农杆菌菌液浓度比为1:1的情况下,农杆菌EHA105/pCAMBIA1300与EHA105/pCAMBIA0301组合共转化水稻的效率要高于其他菌株的组合;在以农杆菌EHA105/pCAMBIA1300与EHA105/pCAMBIA0301进行共转化时,两种菌液浓度比为1:2时共转化效率最高。Two binary vectors, pCAMBIA1300 and pCAMBIA0301 containing hybromycin resistance gene （HPT） and GUS reproter gene in their T-DNA region, respectively, were used to investigate the efficiency of cotransformation of rice. The effects of either the combinations or the densities of Agrobacteriurn strains on co-transformation efficiency were carefully tested according to both the GUS transient expression in infected calli and the stable co-transformation efficiency. The results showed that the combination with Agrobacterium strains EHA105/ pCAMBIA1300 and EHA105/ pCAMBIA0301 achieved a higher co-transformation efficiency than that of the other bacterium combinations when the same density of these two bacteriums was used to infect the rice callus. Additionally, when the density of two Agrobacteriurn strains EHA105/ pCAMBIA1300 and EHA105/ pCAMBIA0301 was adjusted to 1：2, the co-transformation efficiency was higher than that of the other density ratio.

关 键 词：水稻 共转化 T-DNA区 GUS瞬间表达 水稻愈伤组织 农杆菌介导 共转化法 双元载体 GUS报告基因 中部 选择标记基因 转化效率 T-DNA 

分 类 号：S511[农业科学—作物学]