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Background The skilful handling of N fertilizer, including N source type and its timing, is necessary to obtain maximum profitability in wheat crops in terms of production and quality. Studies on grain yield and quality with ammonium as sole N source have not yet been conducted. The aim of this study was to evaluate the effect of N source management (nitrate vs. ammonium), and splitting it into two or three amendments during the wheat life cycle, on grain yield and quality under irrigated conditions. Results This experiment demonstrates that Cezanne wheat plants growing with ammonium as exclusive N source are able to achieve the same yield as plants growing with nitrate and that individual wheat plants grown in irrigated pots can efficiently use late N applied in GS37. Ammonium nutrition increased both types of grain reserve proteins (gliadins and glutenins) and also increased the ratio gli/glu with respect to nitrate nutrition. The splitting of the N rate enhanced the ammonium effect on grain protein composition. Conclusions The application of ammonium N source, especially when split into three amendments, has an analogous effect on grain protein content and composition to applications at a higher N rate, leading to higher N use efficiency. © 2012 Society of Chemical Industry [ABSTRACT FROM AUTHOR]