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Reducing Nitrogen Rate and Increasing Plant Density Accomplished High Yields with Satisfied Grain Quality of Soft Wheat via Modifying the Free Amino Acid Supply and Storage Protein Gene Expression.

In a 2 yr field experiment, we investigated the combined effects of reduced nitrogen (N) rate and increased plant density on the trade-off between the grain protein content (GPC) and the grain yield (GY) in soft wheat cultivars. Reducing N application significantly decreased both GPC and GY; however, to some extent, increasing the top-dressed N ratio and plant density compensated for the GY loss. Optimizing the combination of these three factors (150 kg N ha-1 with 50% top-dressed N and 360 × 104 plants ha-1) achieved both the required lower GPC for soft wheat and relatively higher GY compared with the conventional cultivation strategy. In addition, this optimized combination downregulated 11 high-molecular-weight glutenin subunits, 8 low-molecular-weight glutenin subunits, 5 α/β-gliadins, and 2 γ-gliadins in mature grains as identified by data-independent acquisition mass spectrometry. Further analysis indicated that the relatively lower free amino acid content and downregulated expressions of the seed storage protein (SSP) synthesis-related genes in filling grains contributed to the reduction of SSP and GPC. Furthermore, the dilution effect induced by a relatively higher accumulation of starch than proteins also partially explained the reduced GPC. Unlike proteins, grain starch accumulation and content depended more on the soluble sugar availability, rather than on the starch synthesis capacity. These findings provide novel insights on simultaneous improvement in the grain quality and yield of soft wheat through synchronized manipulations of N fertilization and plant density.