Integration of widely targeted and targeted metabolomics reveals flavonoid accumulation profiles across different cultivars of Ludisia discolor

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Introduction: , a rare orchid species, possesses significant medicinal value; however, the metabolic heterogeneity among its cultivars and the specific accumulation patterns of bioactive flavonoids remain largely uncharacterized. Methods: cultivars (Funongdanxia [DX], Funongruyi [RY], Xigonghong [XGH], Minreyinlong [YL], and Minreyuanshuai [YS]). Results: A total of 2,286 metabolites were identified, from which 802 differential metabolites were identified, with the majority belonging to flavonoids, lipids, and terpenoids. Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) revealed that the metabolic profiles of the XGH and YL cultivars were significantly distinct from the others. KEGG pathway enrichment analysis indicated that phenylpropanoid biosynthesis, flavonoid biosynthesis, and flavone and flavonol biosynthesis are the core pathways driving cultivar differentiation. Additionally, three common differential Kaempferol-4'-O-glucoside*, 1-O-(3,4,5-Trimethoxybenzoyl)-β-D-Glucopyranoside, and sec-o-Glucosylhamaudol were identified as potential key chemical markers for cultivar discrimination. Targeted quantitative analysis further validated that XGH and YL possess a significant advantage in the accumulation of 11 key flavonoid components, with flavonol glycosides exhibiting the most prominent accumulation characteristics. Discussion: cultivars, uncovering the unique advantages of XGH and YL in flavonoid accumulation and their specific pathway regulatory modes. These findings provide a solid scientific foundation and data support for the metabolic characterization of medicinal germplasm resources, the breeding of superior high-flavonoid cultivars, and the precise quality evaluation of medicinal materials.

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