The subtropical peach cultivar UFSun grafted on five different rootstocks ('Flordaguard', 'Barton', 'MP‐29', 'P‐22', and 'Okinawa') was investigated in terms of the pomological and biochemical parameters of the fruit. Significant differences in fruit weight and size, soluble solids content, titratable acidity, and firmness were found among some rootstocks. The fruit length and diameter were different between the 'MP‐29' and other rootstocks. It was also found that firmness of 'UFSun' fruit was affected by the rootstock. The highest firmness value was found when 'UFSun' was grafted on 'Flordaguard'.' 'MP‐29' fruit had the highest soluble solids content, but there were no differences among the other rootstocks. 'UFSun' fruit from trees grafted on 'MP‐29' were smaller and had the most intense color compared to 'UFSun' fruit from other rootstocks. Rootstock had a significant influence on total fruit phenolic compounds, anthocyanin content, and total antioxidant activity, with fruit from 'UFSun' on 'MP‐29' having the highest values in all of them. A high correlation between fruit total antioxidant activity and total phenolic content was found. Overall, the results showed that 'MP‐29' seems to induce the highest fruit quality, showing higher contents of total soluble solids, total titratable acidity, total phenolic compounds, total antioxidant activity, and total anthocyanin content. Selecting the right combination of the rootstock and cultivar is important for optimizing fruit quality parameters.