R2R3-MYB transcription factor MYB113 specifically regulates anthocyanin accumulation in Lycium ruthenicum

Lycium ruthenicum Murray (black goji berry), a Solanaceae medicinal plant, is valued for its high flavonoid content. However, the transcriptional regulation of flavonoid biosynthesis in L. ruthenicum remains unclear, hindering its pharmaceutical development. Here, we identified and characterized LrMYB113 , an R2R3-MYB transcription factor, as a key regulator of anthocyanin biosynthesis in L. ruthenicum . Phylogenetic analysis grouped LrMYB113 into the anthocyanin-associated S6 subgroup of MYBs. Heterologous expression of LrMYB113 in tobacco induced pigment accumulation and upregulated anthocyanin pathway genes. LrMYB113 overexpression in L. ruthenicum hairy roots enhanced accumulation of four acylated anthocyanins and activated anthocyanin pathway genes. Yeast two-hybrid and bimolecular fluorescence complementation assays showed LrMYB113 interacts with bHLHs (LrJAF13/LrAN1b) and WD40 (LrAN11) to form an MBW complex. Promoter binding and transactivation assays demonstrated LrMYB113 directly binds to and activates LrDFR and LrANS promoters. Dual-luciferase assays showed LrMYB113 alone strongly activates LrDFR and LrANS promoters; MBW complexes enhanced activity compared to individual bHLH/WD40 but not to LrMYB113 alone. Our findings identify LrMYB113 as a critical regulator of anthocyanin biosynthesis in L. ruthenicum , shedding light on flavonoid molecular mechanisms and supporting genetic improvement for pharmaceutical use.