Sugar and acid metabolism are critical for fruit ripening and quality formation, but the underlying regulatory mechanisms are largely unknown. Here, we identified a transcriptional repressor, FaMYB44.2, that regulates sugar and acid accumulation in strawberry (Fragaria × ananassa 'Benihoppe') receptacles. We transiently expressed FaMYB44.2 in strawberry fruit and conducted metabolic and molecular analyses to explore the role of FaMYB44.2 in sugar and acid accumulation in strawberry. We found that FaMYB44.2 negatively regulates soluble sugar accumulation and malic acid content and represses the expression of numerous structural genes, including FaSPS3, a key gene in sucrose accumulation. From the white fruit stage onwards, the repressive effect of FaMYB44.2 on FaSPS3 is reversed by FaMYB10, which positively regulates anthocyanin accumulation. Our results indicate that FaMYB10 suppresses FaMYB44.2 expression; weakens the interaction between FaMYB44.2 and its co-repressor, FabHLH3; and cooperates with FabHLH3 to activate the expression of FaSPS3. The interplay between FaMYB10 and FaMYB44.2 results in sucrose accumulation in ripe strawberry fruits. In addition, the repressive effect of FaMYB44.2 on sucrose accumulation is enhanced by jasmonic acid. This study provides new insights into the regulatory mechanisms of sucrose accumulation and sheds light on the interplay between regulatory proteins during strawberry fruit ripening and quality formation.