农杆菌介导的小麦转化体系的优化  

Optimization of genetic transformation of wheat cultivars mediated by Agrobacterium tumefaciens

作　　者：张月婷[1,2] 廖玉才[1,2] 黄涛[1,3] 刘正位[1] 高春生[1] 李和平[1,3] 

机构地区：[1]华中农业大学麦类作物分子生物技术实验室,武汉430070 [2]华中农业大学植物科学技术学院,武汉430070 [3]华中农业大学生命科学技术学院,武汉430070

出　　处：《华中农业大学学报》2012年第1期23-27,共5页Journal of Huazhong Agricultural University

基　　金：教育部高等学校博士学科点专项科研基金(20070504010)和转基因专项(2009ZX08002-001B)

摘　　要：为建立农杆菌介导的高效小麦遗传转化体系,分析比较了不同小麦栽培品种、不同培养处理条件下的愈伤诱导、分化及转化效率。结果表明:2个长江中下游主推小麦品种扬麦158和华麦13的幼胚愈伤诱导没有差异,但扬麦158形成的胚性愈伤多、分化率及转化率高于华麦13,是优良的农杆菌介导的遗传转化受体基因型。预培养20d的愈伤转化率最高,高渗处理可进一步提高转化率。获得的T0、T1代转基因植株,经PCR分析鉴定,表明外源基因已经整合到小麦基因组中。To develop an efficient transformation system for wheat mediated by Agrobacterium tu- mefaciens, comparative analyses of callus induction, regeneration and transformation efficiency were car- ried out by using different elite wheat eultivars and different culture and induction conditions. The re- sults indicated that two wheat elite cultivars,Yangmai 158 and Huamai13, showed no difference in callus induction,but Yangmai 158 formed more embryogenic calli and had a higher efficiency of regeneration and transformation. Yangmai 158 is a good wheat genotype used for Agrobacterium-mediated transfor- mation. Furthermore/different pre-culture duration times were compared and the results showed that 20 days for the pre-culture gave rise to the highest transformation efficiency. With the method established in this study, transgenic To and T1 wheat lines were generated and identified hy PCR analysis. These results provide useful information for Agrobacterium-mediated transformation of elite wheat cultivars.

关 键 词：小麦 农杆菌 幼胚 除草剂 转化体系 

分 类 号：S512.103.53[农业科学—作物学]