Metabolomic Analysis Reveals the Diversity of Defense Metabolites in Nine Cereal Crops.

Cereal crops are important staple foods, and their defense metabolites hold significant research importance. In this study, we employed LC-MS-based untargeted and widely-targeted metabolomics to profile the leaf metabolome of nine cereal species, including rice, wheat, maize, barley, sorghum, common oat, foxtail millet, broomcorn millet, and adlay. A total of 9869 features were detected, among them, 1131 were annotated, encompassing 18 classes such as flavonoids, lipids, and alkaloids. Results revealed that 531 metabolites were detected in all species, while each cereal crop possessed 4 to 12 unique metabolites. Focusing on defense metabolites, we identified eight benzoxazinoids uniquely present in maize, wheat, and adlay. Hierarchical clustering based on metabolite abundance divided all metabolites into nine clusters, and subsequent pathway enrichment analysis revealed that the stress-related flavonoid biosynthesis pathway was enriched in multiple clusters. Further analysis showed that four downstream compounds of HBOA (2-hydroxy-1,4-benzoxazin-3-one) in the benzoxazinoid biosynthesis pathway were enriched in maize. Wheat uniquely accumulated the 4'-methylated product of tricin, trimethoxytricetin, whereas adlay accumulated the tricin precursor tricetin in the flavonoid biosynthesis pathway. In summary, this study elucidates the metabolic diversity in defense metabolites among various cereal crops, providing valuable background information for the improvement of stress resistance in cereal crops.