Pina^m、Pinb^m和Gsp-1~m基因表达载体构建及遗传转化

Construction of Overexpression Vector of Pinb^m、Pina^m、Gsp-1~m and Genetic Transformation

子粒硬度是小麦重要的品质性状之一,对小麦的磨粉及加工品质有重要影响。子粒硬度主要受位于染色体5D短臂上Hardness(Ha)的两个主效基因Puroindoline a(Pina)和Puroindoline b(Pinb)的调控及基因Grain Softness Protein-1(Gsp-1)的影响。从一粒小麦(Triticum monococcum)DV92中克隆基因Pinam、Pinbm和Gsp-1m,并构建以Bar基因为筛选标记的重组质粒PC186-Pinam、PC186-Pinbm、PC186-Gsp-1m,通过基因枪介导法将它们分别转入普通小麦科农199中。经过PCR和RT-PCR鉴定,Pinam、Pinbm和Gsp-1m基因分别获得2株转基因阳性苗,结果表明转入基因能在小麦中表达,但有关转基因小麦的子粒硬度还需进一步验证。本研究完成了单基因在普通小麦中的遗传转化。

Grain hardness is one of the most important characteristics of bread wheat quality and has an important influence on wheat milling and processing quality. Hardness is mainly controlled by Puroindoline genes （Puroindo- line a, Pina and Puroindoline b, Pinb ） with influence from Grain Softness Protein-1 （Gspd）. All three genes are lo- cated on the Ha locus of 5DS. In this study,three genes （Pina^m ,Pinb^m ,Gsp-1^m） were cloned from Triticum mono- coccum and were constructed to PC186 overexpression vector with Bar gene as selection marker. Gene gun method was used to transform the three genes into common wheat cultivar Kenong 199. Finally, two transgenic plants carry- ing with Pina^m ,Pinb^m and Gsp-1^m genes were obtained after PCR and RT-PCR identification. The hardness of the transgenic wheat lines requires further validation. The study successfully transformed the single gene into common wheat.