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Background  Anthocyanins, which have important biological functions and have a beneficial effect on human health, notably account for pigmentation in purple-fleshed sweet potato tuberous roots. Individual regulatory factors of anthocyanin biosynthesis have been identified; however, the regulatory network of anthocyanin biosynthesis in purple-fleshed sweet potato is unclear.    Results  We functionally determined that  IbMYB340  cotransformed with  IbbHLH2  in tobacco and strawberry receptacles induced anthocyanin accumulation, and the addition of  IbNAC56a  or  IbNAC56b  caused increased pigmentation. Furthermore, we confirmed the interaction of IbMYB340 with IbbHLH2 and IbNAC56a or IbNAC56b via yeast two-hybrid and firefly luciferase complementation assays; these proteins could form a MYB340-bHLH2-NAC56a or MYB340-bHLH2-NAC56b transcriptional complex to regulate anthocyanin biosynthesis by binding to the  IbANS  promoter rather than the  IbUFGT  promoter. Furthermore, it was found by a transient expression system in tobacco leaves that  IbMYB44  could decrease anthocyanin accumulation. Moreover, the interaction of IbMYB44 with IbMYB340 and IbNAC56a or IbNAC56b was verified. This result suggested that  IbMYB44  acts as a repressor of anthocyanin in sweet potato.    Conclusions  The repressor  IbMYB44  affected anthocyanin biosynthesis by competitively inhibiting the  IbMYB340 - IbbHLH2 - IbNAC56a  or  IbMYB340 - IbbHLH2 - IbNAC56b  regulatory complex formation. Overall, the present study proposed a novel regulatory network whereby several vital TFs play key roles in regulating anthocyanin biosynthesis, and it provides strong insight into the potential mechanism underlying anthocyanin biosynthesis in sweet potato tuberous roots with purple color.

MYB44 competitively inhibits the formation of the MYB340-bHLH2-NAC56 complex to regulate anthocyanin biosynthesis in purple-fleshed sweet potato