Rice (Oryza sativa) produces phenolamides and diterpenoids as major phytoalexins. Although the biosynthetic pathways of phenolamides and diterpenoids in plants have been revealed, knowledge of their accumulation regulatory mechanisms remains limited, and, in particular, no co-regulatory factor has been identified to date. Here, using a combined co-expression and evolutionary analysis, we identified the basic helix–loop–helix (bHLH) transcription factor OsbHLH5 as a positive bifunctional regulator of phenolamide and diterpenoid biosynthesis in rice. Metabolomic analysis revealed that OsbHLH5 significantly increased the content of phenolamides (such as feruloyl tryptamine (Fer-Trm) and p-coumaroyl tyramine (Cou-Tyr)) and diterpenoid phytoalexins (such as momilactones A, momilactones B) in the overexpression lines, while their content was reduced in the OsbHLH5 knockout lines. Gene expression and dual-luciferase assays revealed that OsbHLH5 activates phenolamide biosynthetic genes (including putrescine hydroxycinnamoyltransferase 3 (OsPHT3), tyramine hydroxycinnamoyltransferases 1/2 (OsTHT1/2), and tryptamine benzoyltransferase 2 (OsTBT2)) as well as diterpenoid biosynthetic genes (including copalyl diphosphate synthase 4 (OsCPS4) and kaurene synthase-like 4/7/10/11 (OsKSL4/7/10/11)). Furthermore, we have demonstrated that OsbHLH5 is induced by jasmonic acid (JA), while pathogen inoculation assays indicated that the overexpression of OsbHLH5 in transgenic rice plants leads to enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo). Overall, we have identified a positive regulator of phenolamide and diterpenoid biosynthesis and have demonstrated that biotic stress induces phytoalexin accumulation partly in an OsbHLH5-dependent manner, providing new insights into the metabolic interactions involved in pathogen response and offering valuable gene resources for the development, through genetic improvement, of new rice varieties that are resistant to diseases.

Wampee (Clausena lansium (Lour.) Skeels) has natural bioactive components with diverse health benefits, but its detailed metabolism and tissue distribution are not fully understood. Here, widely targeted metabolomics analysis methods were employed to analyze the wampee fruit (peel, pulp, and seed) of 17 different varieties. A total of 1286 metabolites were annotated, including lipids, flavonoids, polyphenols, carbazole alkaloids, coumarins, and organic acids, among others. The quantitative analysis and matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI-MSI) analysis indicated remarkable variations in metabolite categories and content in the peel, pulp, and seed of wampee fruit. Additionally, the difference analysis found that the metabolic components of peel contributed dominantly to the differences among varieties, and 7 potential biomarkers were identified. In this study, a comprehensive metabolome landscape of wampee fruit was established, which provided important information for the isolation and identification of functional components, food industry application, and nutritional improvement breeding.

To characterize the nutrients and bioactive compounds in grape berries and to explore the real cause of the “French paradox” phenomenon, we performed metabolomic analysis of 66 grape varieties worldwide using liquid chromatography–tandem mass spectrometry (LC-MS). A nontargeted metabolomics approach detected a total of 4889 metabolite signals. From these, 964 bioactive and nutrient compounds were identified and quantified, including modified flavonoids, medicinal pentacyclic triterpenoids, vitamins, amino acids, lipids, etc. Interestingly, metabolic variations between varieties are not explained by geography or subspecies but can be significantly distinguished by grapes’ color, even after excluding flavonoids and anthocyanins. In our analysis, we found that purple grape varieties had the highest levels of key bioactive components such as flavonoids, pentacyclic triterpenes, and polyphenols, which are thought to have a variety of health benefits such as antioxidant, anti-inflammatory, and antitumor properties, when compared to grapes of other colors. In addition, we found higher levels of vitamins in red and pink grapes, possibly explaining their role in preventing anemia and scurvy and protecting the skin. These findings may be a major factor in the greater health benefits of wines made from purple grapes. Our study provides comprehensive metabolic profiling data of grape berries that may contribute to future research on the French paradox.

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.