Institute of Food Sciences, National Research Council (CNR) , 83100 Avellino, Italy.
Department of Agriculture, University of Naples "Federico II" , 80100 Portici, Italy.
J Agric Food Chem. 2017 Apr 19;65(15):3242-3250. doi: 10.1021/acs.jafc.7b00571. Epub 2017 Apr 5.
The time course of biosynthesis and accumulation of storage proteins in developing grains of durum wheat (Triticum turgidum ssp. durum (Desf.) Husn.) pasta-quality reference cv. Svevo was investigated at the protein level for the first time. Seeds were harvested at key kernel developmental stages, namely, 3 (seed increase 3-fold in size), 5 (kernel development, water-ripe stage), 11 (kernel development, water-ripe stage), 16 (kernel full development, water-ripe stage), 21 (milk-ripe stage), and 30 (dough stage) days postanthesis (dpa). Gliadins and glutenins were fractionated according to their different solubility and individually analyzed after fractionation by reversed-phase high performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were identified by liquid chromatography-tandem mass spectrometry of proteolytic peptides. The α- and γ-gliadin were already detected at 3 dpa. The biosynthesis of high molecular mass glutenin Bx7 was slightly delayed (11 dpa). Most of the gluten proteins accumulated rapidly between 11 and 21 dpa, with a minor further increase up to 30 dpa. The expression pattern of gluten proteins in Triticum durum at the early stages of synthesis provides reference data sets for future applications in crop breeding and growth monitoring.
首次在蛋白质水平上研究了硬质小麦（Triticum turgidum ssp. durum (Desf.) Husn.）面食品质参考品种 Svevo 发育籽粒中贮藏蛋白的生物合成和积累的时间进程。种子在关键的籽粒发育阶段收获，即：3 天（种子大小增加 3 倍）、5 天（籽粒发育，水熟期）、11 天（籽粒发育，水熟期）、16 天（籽粒完全发育，水熟期）、21 天（乳熟期）和 30 天（面团期）授粉后天数（dpa）。醇溶蛋白和麦谷蛋白根据其不同的溶解度进行分级，然后通过反相高效液相色谱和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行分级后单独分析。通过肽的液相色谱-串联质谱鉴定蛋白质。在 3 dpa 时已经检测到 α-和 γ-醇溶蛋白。高分子质量谷蛋白 Bx7 的生物合成略有延迟（11 dpa）。大多数谷蛋白在 11 至 21 dpa 之间迅速积累，在 30 dpa 时略有增加。硬质小麦在合成早期谷蛋白的表达模式为作物育种和生长监测的未来应用提供了参考数据集。
