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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 important quality traits of wheat, wheat milling and processing quality have a significant impact. The grain hardness was mainly affected by the regulation of the two major genes Puroindoline a (Pina) and Puroindoline b (Pinb) located on Hardness (Ha) on the short arm of chromosome 5D and the gene Grain Softness Protein-1 (Gsp-1). Pinam, Pinbm and Gsp-1m were cloned from Triticum monococcum DV92 and recombinant plasmids PC186-Pinam, PC186-Pinbm and PC186-Gsp-1m with Bar gene screening were constructed and gene-mediated France, they were transferred to the common wheat Section 199 agricultural. Two transgenic positive plants were obtained by PCR and RT-PCR. The results showed that the transgene could be expressed in wheat, but the grain hardness of transgenic wheat needs further verification. This study completed the genetic transformation of single gene in common wheat.