Eugenol is a volatile phenylpropanoid that contributes to flower and ripe fruit scent. In ripe strawberry (Fragaria Χ ananassa) fruit receptacles, eugenol is biosynthesized by eugenol synthase (FaEGS2). However, the transcriptional regulation of this process is still unknown. We have identified and functionally characterized a R2R3 MYB transcription factor (FaEOBII) that seems to be the orthologous gene of PhEOBII from Petunia hybrida that contributes to the regulation of eugenol biosynthesis in petals. The expression of FaEOBII was ripe-related and fruit receptacle-specific, although high expression values were also found in petals. This expression pattern of FaEOBII correlated with eugenol content in both fruit receptacle and petals. The expression of FaEOBII was repressed by auxins and activated by abscisic acid (ABA), in parallel to the ripening process. In ripe strawberry receptacles, where the expression of FaEOBII was silenced, FaCAD1 and FaEGS2 expression, two structural genes involved in eugenol production, were down-regulated. A subsequent decrease in eugenol content in ripe receptacles was also observed, confirming the involvement of FaEOBII in eugenol metabolism. Additionally, the expression of FaEOBII was under control of FaMYB10, another R2R3 MYB transcription factor that regulates the early and late biosynthetic genes from the flavonoid/phenylpropanoid pathway. In parallel, the amount of eugenol in FaMYB10-silenced receptacles was also diminished. Taken together, all these data indicate that FaEOBII plays a regulating role of the volatile phenylpropanoid pathway gene expression that gives rise to eugenol production in ripe strawberry receptacles.