Updated project metadata. Drought is one of the most serious abiotic stresses, under which the crop yield is significantly reduced. Elite winter wheat cultivar Henong 341 (Triticum aestivum L.) was grown in experimental fields of China Hebei province (116°37′23″E, 37°41′02″N) Two different treatment conditions were designed: rain-fed (no water irrigation) and well-watered (water irrigation) conditions at jointing and flowering stages. Each treatment included three biological replicates and each plot was 25m2. In this study, a comprehensive phosphoproteomic analysis of developing grains of 26th day after flowering (DAF) in Chinese bread wheat cultivar Henong 341 under well-watered and water-deficit conditions was performed by means of TiO2 enrichment, LC/MS/MS analysis, and Label-free peptide-intensity quantification. Under well-watered conditions, a total of 590 unique phosphopeptides corresponding to 603 nonredundant phosphorylation sites, representing 471 phosphoproteins, were identified. While under water deficit, 63 unique phosphopeptides, corresponding to 61 phosphoproteins, were found to be differently regulated in phosphorylation status compared to well-watered conditions (≥2 fold intensities).