Biostimulant impacts of Glutacetine® and derived formulations (VNT1 and VNT4) on the bread wheat grain proteome.

Nitrogen (N) fertilizer is essential to ensure grain yield and quality in bread wheat. Improving N use efficiency is therefore crucial for wheat grain protein quality. In the present work, we analysed the effects on the winter wheat grain proteome of biostimulants containing Glutacetine® or two derived formulations (VNT1 and 4) when mixed with urea-ammonium-nitrate fertilizer. A large-scale quantitative proteomics analysis of two wheat flour fractions produced a dataset of 4369 identified proteins. Quantitative analysis revealed 9, 39 and 96 proteins with a significant change in abundance after Glutacetine®, VNT1 and VNT4 treatments, respectively, with a common set of 11 proteins that were affected by two different biostimulants. The major effects impacted proteins involved in (i) protein synthesis regulation (mainly ribosomal and binding proteins), (ii) defence and responses to stresses (including chitin-binding protein, heat shock 70 kDa protein 1 and glutathione S-transferase proteins), (iii) storage functions related to gluten protein alpha-gliadins and starch synthase and (iv) seed development with proteins implicated in protease activity, energy machinery, and the C and N metabolism pathways. Altogether, our study showed that Glutacetine®, VNT1 and VNT4 biostimulants positively affected protein composition related to grain quality. Data are available via ProteomeXchange with identifier PXD021513. SIGNIFICANCE: We performed a large-scale quantitative proteomics study of the total protein extracts from flour samples to determine the effect of Glutacetine®-based biostimulants treatment on the protein composition of bread wheat grain. To our knowledge, only a few studies in the literature have applied proteomic approaches to study bread wheat grains and in particular to investigate the effect of biostimulants on the grain proteome of this cereal crop. In addition, most approaches used fractional extraction of proteins to target reserve proteins followed electrophoresis which leads to low identification rate of proteins. We identified and quantified a large protein dataset of 4369 proteins and determined ontological class of proteins affected by biostimulants treatments. Our proteomics investigation revealed the important role of these new biostimulants in achieving significant changes in protein synthesis regulation, storage functions, protease activity, energy machinery, C and N metabolism pathways and responses to biotic and abiotic stresses in grain.