Xu Beiming, Jia Yuku, Feng Jianchao, Yang Yang, Ma Geng, Zhang Yanfei, Xie Yingxin, Ma Dongyun
National Engineering Research Center for Wheat, College of Agronomy, Henan Agricultural University, Zhengzhou 450046, China.
National Wheat Technology Innovation Center, Henan Agricultural University, Zhengzhou 450046, China.
Foods. 2025 Apr 24;14(9):1481. doi: 10.3390/foods14091481.
This study uses proteomic technology to identify differentially expressed proteins (DEPs) under varying nitrogen fertilizer levels. Additionally, it utilizes weighted gene co-expression network analysis (WGCNA) based on expression data of DEP-coding genes to explore the mechanism by which nitrogen promotes grain protein accumulation. The results indicate that high-nitrogen treatment leads to an increased grain protein content, wet gluten content, stability time, and energy area. In addition, the β-sheet content of the protein secondary structure increased, while the irregular curl content decreased. A total of 285 DEPs were identified under different nitrogen levels, with 172 upregulated proteins in grains under high-nitrogen treatment including storage proteins (8.14%) and proteins involved in nitrogen metabolism (8.72%), defense/stress (11.04%), regulation (26.16%), and transport (5.23%). This suggests that both storage proteins and certain metabolic proteins contribute to dough network formation. WGCNA revealed a strong correlation between the blue module and grain samples, and Gene Ontology analysis indicated that most genes were enriched in response to abscisic acid (ABA) in the "biological process" category. Furthermore, 18 core genes were identified, with most containing ABA response elements, light response elements, and motifs related to storage protein regulation in their promoter regions. Expression analysis of 10 genes and their predicted transcription factors during the grain-filling stage demonstrated higher expression levels under high-nitrogen conditions. This study provides valuable insights into the promotion of grain protein accumulation and dough quality by nitrogen fertilizer application.
本研究采用蛋白质组学技术鉴定不同氮肥水平下差异表达的蛋白质（DEP）。此外，基于DEP编码基因的表达数据，利用加权基因共表达网络分析（WGCNA）探索氮促进籽粒蛋白质积累的机制。结果表明，高氮处理导致籽粒蛋白质含量、湿面筋含量、稳定时间和能量面积增加。此外，蛋白质二级结构的β-折叠含量增加，而无规则卷曲含量减少。在不同氮水平下共鉴定出285个DEP，高氮处理下籽粒中有172个上调蛋白，包括贮藏蛋白（8.14%）和参与氮代谢的蛋白（8.72%）、防御/应激（11.04%）、调控（26.16%）和转运（5.23%）。这表明贮藏蛋白和某些代谢蛋白都有助于面团网络的形成。WGCNA显示蓝色模块与籽粒样品之间存在强相关性，基因本体分析表明，在“生物学过程”类别中，大多数基因在脱落酸（ABA）响应中富集。此外，鉴定出18个核心基因，其启动子区域大多含有ABA响应元件、光响应元件和与贮藏蛋白调控相关的基序。对灌浆期10个基因及其预测转录因子的表达分析表明，在高氮条件下表达水平较高。本研究为氮肥施用促进籽粒蛋白质积累和面团品质提供了有价值的见解。
