Role of an endodermis‐specific miR858b‐MYB1L module in the regulation of Taxol biosynthesis in Taxus mairei

Taxol, a chemotherapeutic agent widely used for treating various cancers, is extracted from the stems of Taxus mairei. However, current knowledge regarding the effects of stem tissue and age on Taxol accumulation is limited. We employed matrix-assisted laser desorption/ionization mass spectrometry to visualize taxoids in stem section sections of varying ages from T. mairei. Laser capture microdissection integrated with data-dependent acquisition-MS/MS analysis identified that several Taxol biosynthesis pathway-related enzymes were predominantly produced in the endodermis, elucidating the molecular mechanisms underlying endodermis-specific Taxol accumulation. We identified an endodermis-specific MYB1-like (MYB1L) protein and proposed a potential function for the miR858-MYB1L module in regulating secondary metabolic pathways. DNA affinity purification sequencing analysis produced 92 506 target peaks for MYB1L. Motif enrichment analysis identified several de novo motifs, providing new insights into MYB recognition sites. Four target peaks of MYB1L were identified within the promoter sequences of Taxol synthesis genes, including TBT, DBTNBT, T13OH, and BAPT, and were confirmed using electrophoretic mobility shift assays. Dual-luciferase assays showed that MYB1L significantly activated the expression of TBT and BAPT. Our data indicate that the miR858b-MYB1L module plays a crucial role in the transcriptional regulation of Taxol biosynthesis by up-regulating the expression of TBT and BAPT genes in the endodermis.