| Summary: | Herbaceous peony (<i>Paeonia lactiflora</i> Pall.) is a famous ornamental plant, and the study of its flower color is of great significance for cultivating new flower varieties. To explore the factors driving the formation and change of herbaceous peony flower color, we selected five herbaceous peony varieties at four flowering stages to determine the change in flower color, petal area, and microstructure. We also examined the composition and content of petal pigments, soluble sugar and soluble protein content, pH value of cell fluid, and water content. Finally, we analyzed the correlations between each factor. We found that Pn3G5G, Pg3G5G, and Cy3G5G were the main anthocyanin components in red and purple petals. Qu3G, Qu7G, Is3G, and lutein play important roles in yellow petal formation. The change in herbaceous peony flower color during the flowering process is directly caused by changes in the anthocyanin and carotenoid content in petals. In addition, changes in other physiological indices also influence the change in flower color. This study explored the physiological and biochemical factors affecting the color of herbaceous peony petals, which has an important practical significance for studying the physiological mechanism of herbaceous peony flower color formation and color breeding of new flowers.
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