The PpERF3 promotes ethylene-induced anthocyanin biosynthesis in peach

Ethylene is a key hormone regulating fruit ripening; however, the mechanism by which it controls anthocyanin biosynthesis during peach fruit ripening remains a pivotal question in the field. In this study, physiological observations showed that exogenous ethylene treatment influences anthocyanin accumulation, but the specific molecular mechanism underlying this process remains unclear. Transcriptome analysis of peach mesocarp from red-fleshed and white-fleshed varieties at different developmental stages, combined with exogenous ethylene treatment and RT-qPCR, revealed that the ethylene response factor PpERF3 is likely involved in anthocyanin biosynthesis. Overexpression of PpERF3 in tobacco leaves and peach exocarp significantly induced coloration. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays confirmed that PpERF3 interacts with PpMYB10.1, and that the PpMYB10.1-bHLH3 module activates the transcription of anthocyanin biosynthesis genes PpCHI/F3H/DFR/UFGT. Furthermore, yeast one-hybrid (Y1H) and dual-luciferase (Dual-LUC) assays demonstrated that PpERF3 activates the transcription of PpCHI/F3H/DFR/UFGT/MYB10.1, thereby promoting anthocyanin biosynthesis. This study demonstrates that PpERF3 promotes anthocyanin accumulation during peach ripening through direct gene activation or its involvement in a transcription factor complex, uncovering a synergistic ERF-MYB regulatory mechanism for fruit coloration.