Flavonols are important ultraviolet light protectants in many plants and contribute substantially to the quality and healthpromoting effects of fruits and derived plant products. To study the regulation of flavonol synthesis in fruit, we isolated and characterized the grapevine (Vitis vinifera 'Shiraz') R2R3-MYB transcription factor VvMYBF1. Transient reporter assays established VvMYBF1 to be a specific activator of flavonol synthase1 (VvFLS1) and several other promoters of grapevine and Arabidopsis (Arabidopsis thaliana) genes involved in flavonol synthesis. Expression of VvMYBF1 in the Arabidopsis mutant myb12 resulted in complementation of its flavonol-deficient phenotype and confirmed the function of VvMYBF1 as a transcriptional regulator of flavonol synthesis. Transcript analysis of VvMYBF1 throughout grape berry development revealed its expression during flowering and in skins of ripening berries, which correlates with the accumulation of flavonols and expression of VvFLS1. In addition to its developmental regulation, VvMYBF1 expression was light inducible, implicating VvMYBF1 in the control of VvFLS1 transcription. Sequence analysis of VvMYBF1 and VvFLS1 indicated conserved putative light regulatory units in promoters of both genes from different cultivars. By analysis of the VvMYBF1 amino acid sequence, we identified the previously described SG7 domain and an additional sequence motif conserved in several plant MYB factors. The described motifs have been used to identify MYB transcription factors from other plant species putatively involved in the regulation of flavonol biosynthesis. To our knowledge, this is the first functional characterization of a light-inducible MYB transcription factor controlling flavonol synthesis in fruit.

Previous results indicated that in grapevine (Vitis vinifera), regulation of the flavonoid pathway genes by MYB transcription factors depends on their interaction with basic helix-loop-helix proteins (bHLHs). The present study describes the first functional characterization of a bHLH factor from grapevine named VvMYC1. This transcription factor is phylogenetically related to Arabidopsis bHLH proteins, which participate in the control of flavonoid biosynthesis and epidermal cell fate. Transient promoter and yeast two-hybrid assays demonstrated that VvMYC1 physically interacts with MYB5a, MYB5b, MYBA1/A2, and MYBPA1 to induce promoters of flavonoid pathway genes involved in anthocyanin and/or proanthocyanidin (PA) synthesis. Additionally, transient promoter assays revealed that VvMYC1 is involved in feedback regulation of its own expression. Transcript levels of VvMYC1 during berry development correlate with the synthesis of anthocyanins and PAs in skins and seeds of berries, suggesting that VvMYC1 is involved in the regulation of anthocyanins and PA synthesis in these organs. Likewise, transient expression of VvMYC1 and VvMYBA1 induces anthocyanin synthesis in grapevine suspension cells. These results suggest that VvMYC1 is part of the transcriptional cascade controlling anthocyanin and PA biosynthesis in grapevine.