| Summary: | Wheat is one of the most important cereals for human nutrition, but nitrogen (N) losses during its cultivation cause economic problems and environmental risks. In order to improve N use efficiency (NUE), biostimulants are increasingly used. The present study aimed to evaluate the effects of Glutacetine<sup>®</sup>, a biostimulant sprayed at 5 L ha<sup>−1</sup> in combination with fertilizers (urea or urea ammonium nitrate (UAN)), on N-related traits, grain yield components, and the grain quality of winter bread wheat grown at three field sites in Normandy (France). Glutacetine<sup>®</sup> improved grain yield via a significant increase in the grain number per spike and per m<sup>2</sup>, which also enhanced the thousand grain weight, especially with urea. The total N in grains and the NUE tended to increase in response to Glutacetine<sup>®</sup>, irrespective of the site or the form of N fertilizer. Depending on the site, spraying Glutacetine<sup>®</sup> can also induce changes in the grain ionome (analyzed by X-ray fluorescence), with a reduction in P content observed (site 2 under urea nutrition) or an increase in Mn content (site 3 under UAN nutrition). These results provide a roadmap for utilizing Glutacetine<sup>®</sup> biostimulant to enhance wheat production and flour quality in a temperate climate.
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