Planting density is important factor which influence yield and quality of wheat (Triticum aestivum L.) For this reason, in scientific investigations is constantly investigated optimization of plant number per unit area. The objective of this study was to determine the influence of seeding rate in grain quality of winter wheat cultivars. The experiment was conducted with four winter wheat genotypes (’Ana Morava’, ’Vizija’, ’L-3027’, and ’Perla’) at the Small Grains Research Centre of Kragujevac, Serbia, in 3 yr at two seeding rates (SR1 = 500 and SR2 = 650 germinating seeds m-2). The 1000-kernel weight, Zeleny sedimentation, and wet gluten content in divergent wheat genotypes were investigated depending on the seeding rate and ecological factors. Significant differences in quality components were established between investigated seeding rates. The highest values of all investigated quality traits were established in SR2 variant when applied 650 seeds m-2. Genotypes reacted differently to seeding rate. ’Perla’ in average had the highest mean sedimentation value (42.2 mL) and wet gluten content (33.76%) in SR2 variant and this cultivar responded the best to seeding rate. Significant differences for sedimentation value and wet gluten content were found among cultivars, years, seeding rate, and for all their interactions. Also, ANOVA for 1000-kernel weight showed highly significant differences among investigated varieties, seeding rate and growing seasons, but all their interactions were not significant. In all investigated genotypes, better quality was established in SR2 variant when applied 650 seeds m-2.