Temporospatial pattern of flavonoid metabolites and potential regulatory pathway of PbMYB211-coordinated kaempferol-3-O-rhamnoside biosynthesis in Phoebe bournei

Phoebe is a well-known timber tree species that contains abundant metabolites characterized by flavonoids that are widely used in the pharmaceutical industry. Nevertheless, temporospatial flavonoid metabolism variations substantially impact the Phoebe industry. Thus, a metabolomics analysis was carried out and identified 465 metabolites (102 flavonoids) in P. bournei, revealing distinct distribution patterns among five studied organs, and most of the flavonoids were dominant in the leaves. Furthermore, three kaempferol glycoside derivatives were significantly accumulated in the leaves and showed higher contents in young leaves than in mature leaves and differences between spring and autumn. For instance, greater accumulation of kaempferol-7-O-rhamnoside was detected in spring, whereas higher contents of kaempferol-3-O-arabinofuranoside and kaempferol-3-O-rhamnoside were found in autumn. Integrated metabolomics and transcriptomics identified 20 transcription factors (TFs) and 12 structural genes that participate in kaempferol derivative synthesis and elucidated a potential regulatory mechanism in P. bournei. Of the identified genes, PbMYB211 might contribute significantly to the kaempferol-3-O-rhamnoside content by regulating the target structural gene PbUGT139, as revealed by transient overexpression analysis. Overall, this study illuminated the temporospatial accumulation of flavonoids among different organs, seasons, and developmental stages in P. bournei and elucidated a potential regulatory pathway of kaempferol-3-O-rhamnoside. The results provide important insights into harvest techniques and a theoretical basis for the comprehensive utilization of P. bournei.