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Background: Nymphaea atrans exhibits a gradual flower color transition from nearly white to rose-red during anthesis, yet the molecular mechanisms of this phenomenon remain unclear. In the present study, transcriptomic and metabolomic analyses were performed to systematically investigate anthocyanin accumulation patterns and regulatory mechanisms during the color transition of N. atrans. Methods: Petals were collected at three flowering stages: day 1 (D1), day 3 (D3), and day 5 (D5). Targeted metabolomics was performed using UPLC-ESI-MS/MS to profile anthocyanin and other flavonoid metabolites. Transcriptome analysis was conducted via RNA-seq. Differentially accumulated metabolites (DAMs) and differentially expressed genes (DEGs) were identified, followed by functional enrichment and integration analysis. Results: The results revealed significant accumulation of seven anthocyanins, including cyanidin-3-O-arabinoside, cyanidin-3-O-glucoside, cyanidin-3-O-galactoside, cyanidin-3-O-(6″-O-acetyl)-glucoside, at stages D3 (day 3 after flowering, light pink petals) and D5 (day 5 after flowering, deep pink petals), accompanied by the upregulation of key enzyme-encoding genes, chalcone synthase, chalcone isomerase, flavanone 3-hydroxylase, di-hydroflavonol 4-reductase, and anthocyanidin synthase in the anthocyanin biosynthetic pathway. Genes involved in JA biosynthesis and key regulatory genes in the JA signaling pathway were significantly up-regulated, indicating that the JA signaling pathway may play an important regulatory role in the synthesis of anthocyanins in N. atrans. Conclusions: This study unravels the metabolic and molecular underpinnings of flower color transition in N. atrans, thereby establishing a theoretical basis for the targeted regulation of floral pigmentation and molecular breeding of ornamental water lilies. Full article