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Summary  Anthocyanins have crucial biological functions and affect quality of horticultural produce. Anthocyanins accumulate in ripe peach fruit; differential accumulation is observed in deep coloured cultivar ‘Hujingmilu’ and lightly pigmented cultivar ‘Yulu’. The difference was not fully explained by accumulation of total flavonoids and expression of anthocyanin biosynthetic genes. Expression analysis was conducted on a  glutathione S‐transferase  gene ( PpGST1 ), and it was found that the expression correlated well with anthocyanin accumulation in peach fruit tissues. Functional complementation of the  Arabidopsis tt19  mutant indicated that PpGST1 was responsible for transport of anthocyanins but not proanthocyanidins. PpGST1 was localized in nuclei and the tonoplast, including the sites at which anthocyanin vacuolar sequestration occurred. Transient overexpression of  PpGST1  together with  PpMYB10.1  in tobacco leaves and peach fruit significantly increased anthocyanin accumulation as compared with  PpMYB10.1  alone. Furthermore, virus‐induced gene silencing of  PpGST1  in a blood‐fleshed peach not only resulted in a reduction in anthocyanin accumulation but also a decline in expression of anthocyanin biosynthetic and regulatory genes.  Cis ‐element analysis of the  PpGST1  promoter revealed the presence of four MYB binding sites (MBSs). Dual‐luciferase assays indicated that PpMYB10.1 bound to the promoter and activated the transcription of  PpGST1  by recognizing MBS1, the one closest to the ATG start codon, with this trans‐activation being stronger against the promoter of deep coloured ‘Hujingmilu’ compared with lightly coloured cultivar ‘Yulu’. Altogether, our data provided molecular evidence supporting coordinative regulatory roles of PpGST1 and PpMYB10.1 in anthocyanin accumulation in peach.

PpGST1 , an anthocyanin‐related glutathione S‐transferase gene, is essential for fruit coloration in peach