The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions. This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis, gluten secondary structure, and breadmaking quality. Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments. Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation, and reduced flour moisture. In particular, high N-fertilizer application increased β-sheet content by 10.4% and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%. These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality, including superior loaf internal structure, volume, and score. Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation, gluten secondary structure formation, and baking quality.

Nitrogen (N), a macronutrient essential for plant growth and development, is needed for biosynthesis of protein and starch, which affect grain yield and quality. Application of high-N fertilizer increases plant growth, grain yield, and flour quality. In this study, we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and two-dimensional difference gel electrophoresis (2D-DIGE). Application of high-N fertilizer led to significant increases in gluten macropolymer content, total gliadin and glutenin content, and the accumulation of individual storage protein components. Of 126 differentially accumulated proteins (DAPs) induced by high-N conditions, 24 gliadins, 12 high-molecular-weight glutenins, and 27 low-molecular-weight glutenins were significantly upregulated. DAPs during five kernel developmental stages displayed multiple patterns of accumulation. In particular, gliadins and glutenins showed respectively five and six accumulation patterns. The accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.

Wheat seed development is an important physiological process of seed maturation and directly affects wheat yield and quality. In this study, we performed dynamic transcriptome microarray analysis of an elite Chinese bread wheat cultivar (Jimai 20) during grain development using the GeneChip Wheat Genome Array. Grain morphology and scanning electron microscope observations showed that the period of 11–15 days post-anthesis (DPA) was a key stage for the synthesis and accumulation of seed starch. Genome-wide transcriptional profiling and significance analysis of microarrays revealed that the period from 11 to 15 DPA was more important than the 15–20 DPA stage for the synthesis and accumulation of nutritive reserves. Series test of cluster analysis of differential genes revealed five statistically significant gene expression profiles. Gene ontology annotation and enrichment analysis gave further information about differentially expressed genes, and MapMan analysis revealed expression changes within functional groups during seed development. Metabolic pathway network analysis showed that major and minor metabolic pathways regulate one another to ensure regular seed development and nutritive reserve accumulation. We performed gene co-expression network analysis to identify genes that play vital roles in seed development and identified several key genes involved in important metabolic pathways. The transcriptional expression of eight key genes involved in starch and protein synthesis and stress defense was further validated by qRT-PCR. Our results provide new insight into the molecular mechanisms of wheat seed development and the determinants of yield and quality.