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

根据小皮Pin a，Pin b，GSP基㈥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Aegilopstriumialis的基因组DNA和胚乳RNA进行Pin a，Pin b，CSP基因扩增、克降、序列测定和表达分析，发现了两个新型Pin a等位堆因、一个新刑Pin b等化基因和一个新型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，该材料巾含有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．研究结果显示由羊草可以为小麦分子育种提供有用的遗传资源。

Cloning and Expression Analysis of Grain Hardness Controlling Genes in Aegilops triuncialis

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.