Metabolite profiling in ten bread wheat (Triticum aestivum L.) genotypes in response to drought stress

■ Drought-adapted wheat genotypes can be selected by integrating yield components and metabolite profiles. ■ Phenolic acids, carbohydrates, and fatty acids were substantially altered under drought stress. ■ Metabolites such as 1-O-caffeoyl-3-O-p-coumaroylglycerol, 3-beta-3-lupanol and epigallocatechin gallate can be used as biomarkers for drought tolerance. Wheat is frequently constrained by extreme environmental conditions such as drought. Improving drought tolerance in wheat genotypes is crucial for ensuring food security, especially considering the challenges posed by climate change. To reveal the involvement of metabolites in drought response, ten diverse wheat genotypes were investigated under control and water scarcity conditions. The field experiments were set-up, using a 5 × 2 alpha lattice design, with two replicates per treatment, in the 2022 and 2023 growing seasons. Metabolites associated with drought tolerance were analysed using ultra-high performance liquid chromatography, coupled with a quadruple time of flight mass spectrometry (UHPLC-qTOF-MS). Multivariate statistical analysis (MVDA) tools, viz. principal component analysis (PCA) and the orthogonal projection to latent structures-discriminant analysis (OPLS-DA) loading scatter plot were used to identify the metabolites that are positively and negatively correlated to drought stress. Significant variation ( p < 0.05) among genotypes was observed, with 58 metabolites annotated, including phenolic acids, carbohydrates, and fatty acids. The annotated compounds were linked to thirteen most significant pathways, with one carbon metabolism, cutin, suberin and wax synthesis and starch and sucrose metabolism being significantly affected by water stress, based on the KEGG pathway analysis. The two high-yielding wheat genotypes (LM48 and BW140) under drought stress displayed significant upregulation of key metabolites such as sinapoyl hydroxyagmatine, 7-oxostigmasterol, 1-O-caffeoyl-3-O-p-coumaroylglycerol, and 3-beta-3-lupanol, when compared to the non-stressed conditions. This study demonstrates the prospects of applied metabolomics for chemotaxonomic classification, phenotyping and selection in plant breeding, as well as potential use in crop improvement.