蛋白质质量是小麦加工品质改良的关键目标。本项目拟采用双向电泳、高效液相色谱、差异荧光双向电泳和生物质谱等蛋白质组学技术，结合面筋强度和延展性等品质参数分析，对高低分子量亚基组成背景相同、面筋品质差异很大的近等基因系进行谷蛋白、醇溶蛋白、清蛋白和球蛋白等蛋白质组学分析，目的在于：1）鉴定与面筋品质密切相关的目标蛋白质，并克隆其编码基因；2）从转录与翻译水平研究目标蛋白编码基因在籽粒不同发育阶段的表达与调控特征，从蛋白质组成与表达量两个方面探讨面筋品质形成的分子机制；3）构建高效表达载体在E. coli中表达目标蛋白编码基因，回收并纯化表达蛋白，采用实验室体外配粉方法验证其编码基因的功能；4）根据所克隆目标蛋白编码基因的SNP、InDel等变异特征，开发功能标记，通过对现有优质强筋和品质较差品种的分析，验证其有效性；从而为高效改良普通小麦的面筋品质提供理论依据和分子标记。

Gluten quality plays a key role in the improvement of wheat processing quality. Proteomic techniques, including two-dimensional gel electrophoresis (2-DE),high performance liquid chromatography (HPLC), two-dimensional difference gel electrophoresis (2-D DIGE), and matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), will be used to characterize the protein profiles between near-isogenic lines, with the same genetic background in combination of high- and low- molecular weight glutenin subunits, with the objectives to: 1) identify the target proteins closely related with gluten quality, and clone their coding genes using the full length with N-terminal amino acid sequence, with the assistance of conventional polymerase chain reaction (PCR); 2) characterize the gene expression on the samples at five grain developmental stages during the post-anthesis periods, corresponding to the cumulative daily average temperatures, to provide insights into biochemical and mechanisms for key grain protein development from qualitative and quantitative characteristics; 3) purify the protein expressed in E. coli with the coding genes of the target protein introduced, to verify the function of the coding genes using flour blending method; and 4) develop the molecular markers based on the SNPs and InDels among allelic variants of the coding genes, and validate the markers using cultivars with good and poor gluten qualities. The applicability of this approach in proteomics will more widely provide important genes and molecular markers for improvement of wheat gluten quality in breeding programs.