小麦隐性核不育保持系表达载体的构建及拟南芥遗传转化

利用小麦杂种优势能够创制稳定的雄性不育系。相比于细胞质雄性不育系和光温敏雄性不育系，细胞核雄性不育具有育性稳定、易恢复、不受环境影响等特点。小麦隐性核不育基因 ms1的克隆以及杂交制种技术（hybrid seed production technology，SPT）的开发，为雄性不育的保持和杂种优势利用奠定了基础。本研究将小麦花粉育性恢复基因 Ms1、花粉致死基因 ZmAA1、红色荧光蛋白基因 DsRed2及其特异性启动子和终止子连接到表达载体pGEII上，转化野生型拟南芥，得到6株转基因拟南芥。通过荧光显微镜观察拟南芥种子，发现野生型拟南芥种子表面平滑，不能发出红色荧光；而转基因种子表面呈现颗粒状，能够发出红色荧光。这说明表达载体构建成功，并能够在拟南芥中成功表达。这为创制人工小麦隐性核不育系奠定了理论和材料基础。

Expression Vector Construction of Recessive Genic Male Sterile Maintainer Line in Wheat(Triticum aestivum L.) and Genetic Transformation in Arabidopsis

The key to the utilization of heterosis in wheat is to breed stable male sterile lines. Compared with cytoplasmic male sterile lines and photothermal-sensitive male sterile lines,nuclear male sterile lines have the characteristics of stable fertility,easy recovery,and no environmental influences. The cloning of the wheat recessive genic male sterile gene ms1,combined with hybrid seed production technology (hybrid seed production technology,SPT) has laid a foundation for the maintenance of male sterility and the utilization of heterosis. In this study,wheat fertility restoration gene Ms1,pollen lethality gene ZmAA1,red fluorescent protein gene DsRed2 and its specific promoter and terminator were constructed to expression vector pGEII to transform wild-type Arabidopsis thaliana,and 6 transgenic plants were obtained. It was found that the surface of wild-type Arabidopsis seeds was smooth,which can not emit red fluorescence,while the surface of transgenic seeds is granular,which can emit red fluorescence. It showed that the expression vector is successfully constructed and can be expressed in Arabidopsis. This study laid a theoretical and material foundation for the creation of artificial wheat recessive genic male sterile lines.