For the baking quality of wheat flours, the composition and concentration of grain protein are crucial. It is common practice to use late nitrogen (N) application to increase grain protein concentration (GPC) and hence, improve baking quality. However, the use of N fertilizer—particularly shortly before harvest—involves environmental risks. With the suitability of GPC as a parameter for baking quality predictions being more and more questioned, there are further investigations needed considering not only the GPC but also the composition of grain protein. Gluten protein composition varies depending on genotype and environmental factors, such as weather conditions and fertilization rate. To examine whether the effect of a split N application varies under different amounts of total N supply, this study investigates the effects of split nitrogen application on grain protein concentration and composition of wheat (Triticum aestivum L. cv. JB Asano) at four different N fertilization levels (0.8; 1.0; 1.2, and 1.4 g N pot−1) in a pot experiment. The GPC was affected by both, N fertilization level and split N application. In this experiment, the minimum GPC of 13%, which is required for class A wheat varieties, was only achieved when N supply was moderate (at least 1.2 g N per pot). Considering the storage protein composition, the split N application influenced the proportion of α‐/β‐gliadins and γ‐gliadins, the alterations being inconsistent. The ratio of high molecular weight (HMW) to low molecular weight (LMW) glutenin subunits was increased by the split N application only at the lowest N fertilization level. It is concluded that splitting N fertilization into three doses and hence applying one dose of N late in the season can still be a useful approach to improve GPC as well as protein composition – especially when the total N supply is low.
