TCP15 and PIF4 Target the Sulfopeptide-Encoding Gene GOLVEN1/CLE-LIKE6 to Repress Anthocyanin Accumulation

Anthocyanins are protective pigments synthesised by plants to cope with several stressful situations. Anthocyanin synthesis is tightly controlled by multiple transcriptional mechanisms involving the action of activators and repressors. In this work, we report that the class I TCP transcription factor TCP15 and the Phytochrome interacting factor PIF4 negatively affect anthocyanin synthesis by directly inducing the expression of the gene encoding the sulfopeptide GOLVEN1/CLE-LIKE6 (GLV1/CLEL6), a recently identified repressor of anthocyanin synthesis in Arabidopsis thaliana. These transcription factors bind to a region of the GLV1/CLEL6 promoter containing nearby TCP-box and G-box motifs and are able to activate the expression of a reporter gene located under the control of the GLV1/CLEL6 promoter, supporting a direct regulation. Moreover, PIF4 binding to the GLV1/CLEL6 promoter is compromised in a double mutant of TCP15 and the related TCP gene TCP14, whereas TCP15 activation of the GLV1/CLEL6 promoter depends on PIF function, revealing a functional interdependence between these transcription factors. Furthermore, we found that TCP15 and PIF4 also participate in gibberellin-dependent repression of anthocyanin biosynthesis, acting at different levels but independently of GLV1/CLEL6. Altogether, our results add new information on the molecular mechanisms involved in the regulation of anthocyanin accumulation in plants.