To explore the regulation of MADS-box family transcription factors on carotenoid metabolism in yellow fleshed peach, two MADS genes named as PpMADS2(PRUPE_5G208500)and PpMADS3(PRUPE_5G208400)were cloned. The open reading frames of PpMADS2 and PpMADS3 were 768 bp and 756 bp,encoding 255 and 251 amino acids, respectively. Bioinformatics analysis revealed that the N-terminus of both proteins contained typical characteristic domains of MADS-box family. Subcellular localization showed that both PpMADS2 and PpMADS3 were located at the nucleus. The results showed the gene expression levels of PpMADS2,PpMADS3 as well as carotenoid biosynthetic genes such as PpPSY and PpCHYB increased during fruit ripening,which was consistent with the carotenoid accumulation in yellow fleshed peach. Furthermore, the results demonstrated that PpMADS2 could interact with PpMADS3 then synergistically activate the promoters of PpPSY and PpCHYB genes. Therefore,it is speculated that PpMADS2 and PpMADS3 promoted the accumulation of carotenoids in yellow fleshed peach fruit cooperatively through transcriptional activation of PpPSY and PpCHYB. These research provided a theoretical basis for further exploring the regulatory mechanism of MADS-box transcription factors on carotenoid accumulation in peach fruit.