Abstract
BACKGROUNDS: Paphiopedilum orchids, particularly the Chinese endemic Paphiopedilum armeniacum, are prized for their commercial and ornamental value, with the latter serving as a vital breeding resource owing to its distinctive yellow petals. However, the molecular mechanisms underlying yellow petal formation remain unclear. RESULTS: This work employed an integrated transcriptomic and metabolomic comparative analysis of P. armeniacum and two lighter-colored Paphiopedilum species (The sepals and petals are white.) to identify carotenoid-related differentially expressed genes and metabolites before and after blooming in all three species. Metabolomic analysis revealed a marked increase in six differential metabolites, including zeaxanthin, precorrin 2, and β-D-gentiobiosyl crocetin, in P. armeniacum, highlighting their critical role in yellow petal formation. Transcriptomic comparison identified 40 DEGs (including D27, GDSL-like, CYP97B3, LUT1, and PSY) linked to yellow pigmentation, most of which were consistently upregulated in P. armeniacum before and after blooming Integrative metabolomic and transcriptomic analyses demonstrated significant correlations between these genes and metabolites, suggesting their role in regulating carotenoid synthesis and accumulation in yellow petal formation. Furthermore, qRT-PCR elucidates the expression levels of candidate genes, identifying RPL13AD as the optimal reference gene across these three orchid species. CONCLUSIONS: These works elucidate the expression patterns and regulatory roles of carotenoid-related genes in metabolic pathways during P. armeniacum blooming, providing new insights into the molecular mechanisms of carotenoid-mediated plant coloration.