AbstractAnthocyanins are valuable compounds in red-fleshed apples. The MdMYB10 transcription factor is an important regulator of the anthocyanin synthesis pathway. However, other transcription factors are key components of the complex network controlling anthocyanin synthesis and should be more thoroughly characterized. In this study, we used a yeast-based screening technology to identify MdNAC1 as a transcription factor that positively regulates anthocyanin synthesis. The overexpression ofMdNAC1in apple fruits and calli significantly promoted the accumulation of anthocyanins. In binding experiments, we demonstrated that MdNAC1 combines with the bZIP-type transcription factorMdbZIP23to activate the transcription ofMdMYB10andMdUFGT. Our analyses also indicated that the expression ofMdNAC1is strongly induced by ABA because of the presence of an ABREcis-acting element in its promoter. Additionally, the accumulation of anthocyanins in apple calli co-transformed withMdNAC1andMdbZIP23increased in the presence of ABA. Therefore, we revealed a novel anthocyanin synthesis mechanism involving the ABA-induced transcription factor MdNAC1 in red-fleshed apples.