钩刺山羊草Aegilops triuncialis籽粒硬度主效基因的克隆与表达

钩刺山羊草Aegilops triuncialis籽粒硬度主效基因的克隆与表达 Cloning and Expression Analysis of Grain Hardness Controlling Genes in Aegilops triuncialis 陈明洁 CHEN Ming-jie~1 1 first-author 方倜 FANG Ti~2 2 常俊丽 CHANG Jun-li~1 1 杨广笑 YANG Guang-xiao~1 1 何光源 HE Guang-yuan~1 1 华中科技大学生命科学与技术学院 华中科技大学生命科学与技术学院 College of Life Science and Technology,Huazhong University of Science and Technology College of Life Science and Technology,Huazhong University of Science and Technology 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences,Wuhan University College of Life Sciences,Wuhan University 根据小麦Pin a,Pin b,GSP基因的保守序列,设计合成了3对特异性引物,对四倍体钩刺山羊草Ae-gilops triuncialis的基因组DNA和胚乳RNA进行Pin a,Pin b,GSP基因扩增、克隆、序列测定和表达分析,发现了两个新型Pin a等位基因、一个新型Pin b等位基因和一个新型GSP等位基因,基因序列均与六倍体小麦的同源基因存在较大的差异.Southern Blot分析结果表明,该材料中含有2个拷贝Pin a基因,5个拷贝Pin b基因,2个拷贝GSP基因.RT-PCR和Western Blot都证实了Pin a,Pin b,GSP基因在籽粒胚乳中的表达.研究结果显示山羊草可以为小麦分子育种提供有用的遗传资源. According to the conserved regions of Pin a,Pin b and GSP genes,three pairs of specific primers were designed and used to amplify Pin a,Pin b and GSP from young leaves genomic DNA and developing endosperm RNA of tetraploid Aegilops triuncialis.Through gene cloning and sequencing,two new allelic forms of Pin a,one new allelic form of Pin b and GSP have been characterized.Large differences of sequences of Pin a,Pin b and GSP genes existed between Ae.triuncialis and Triticum aestivum.Southern Blot analysis showed that there were two copies of Pin a,five copies of Pin b and two copies of GSP genes in this species.The expression of the three genes in grain endosperm has been identified both by RT-PCR and Western Blot analysis.The results implied that Aegilops provides a useful genetic resource in wheat molecular breeding. 钩刺山羊草 小麦吲哚蛋白 籽粒软质蛋白 籽粒硬度 Aegilops triuncialis puroindolines grain softness protein grain hardness S512.1 国家重点基础研究发展计划（973）资助项目（2002CB111302） 2005-06-01 2021-04-01 6