The sequence and expression analysis of anthocyanin synthase (<i>ANS</i>) genes in <i>Nymphaea colorata</i> with different flower colors

Anthocyanins, a subclass of flavonoids, are key pigments responsible for diverse flower colors in plants. While the role of anthocyanin synthase (<italic>ANS</italic>) genes in regulating anthocyanin accumulation is well established in model plants, their evolutionary diversification and expression patterns in <italic>Nymphaea colorata</italic> remain largely unexplored. This study systematically identified 32 <italic>ANS</italic> genes (<italic>NcANS</italic>) from the <italic>N. colorata</italic> genome and analyzed their physicochemical properties, gene structures, evolutionary relationships, promoter <italic>cis</italic>-acting elements, and expression patterns across blue, red, and white petals. Synteny analysis demonstrated conserved and species-specific expansion of <italic>ANS</italic> genes across plant species. Phylogenetic analysis grouped these genes into seven subfamilies, suggesting functional diversification. Expression profiling and RT-qPCR validation revealed specific <italic>NcANS</italic> genes with higher expression in blue petals, highlighting their potential roles in pigment accumulation. These findings provide critical insights into anthocyanin biosynthesis in <italic>N. colorata</italic> and lay a foundation for future molecular breeding aimed at flower color modification.