Yeast One-Hybrid Screening for Transcription Factors of IbbHLH2 in Purple-Fleshed Sweet Potato

The transcription factor IbbHLH2 has been identified as involved in the biosynthesis of anthocyanins in purple-flesh sweet potatoes. However, little is known about the upstream transcription regulators of the promoter of IbbHLH2 in terms of their involvement in anthocyanin biosynthesis. For this study, the transcription regulators of the promoter of IbbHLH2 were screened via yeast one-hybrid assays in purple-fleshed sweet potato storage roots. Seven proteins, namely IbERF1, IbERF10, IbEBF2, IbPDC, IbPGP19, IbUR5GT, and IbDRM, were screened as upstream binding proteins of the promoter of IbbHLH2. The interactions between the promoter and these upstream binding proteins were verified using dual-luciferase reporter and yeast two-hybrid assays. Furthermore, the gene expression levels of transcription regulators, transcription factors, and structural genes involved in the anthocyanin biosynthesis of different root stages of purple and white-fleshed sweet potatoes were analyzed via real-time PCR. The obtained results indicate that IbERF1 and IbERF10 are key transcription regulators of the promoter of IbbHLH2 and are involved in anthocyanin biosynthesis in purple-fleshed sweet potatoes.