Introduction: Acerola (Malpighia emarginata) fruit is known as one of the richest natural sources of ascorbic acid (vitamin C), containing up to 100 times the amount found in oranges and lemons. This study investigates the seasonal and genotypic effects on fruit quality traits, bioactive compounds, and antioxidant activity in acerola. Methodology: Three pre-selected acerola cultivars (BRS Rubra, Cabocla and Costa Rica) and one commercial cultivar (Junko) were produced under irrigation in semi-arid climate conditions. The fruits were harvested at the red, ripe maturity stage during the spring (October 2022) and summer (February 2023) growing seasons. The fruits were assessed for skin color, soluble solid content (SSC), titratable acidity (TA), SSC/TA ratio, ascorbic acid, phenolic compounds, and antioxidant activity using both 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Results: Acerola quality traits were jointly regulated by the growing season and genotype. However, the genotypic effect was much greater than the seasonal effect for bioactive compounds and antioxidant activity. ‘BRS Rubra’ acerolas stood out in terms of SSC and the SSC/TA ratio, as well as in terms of having low TA, which is important for the perception of fruit sweetness. ‘Junko’ acerolas presented the highest content of bioactive compounds and antioxidant activity. Conclusion: In conclusion, this study provides valuable insights into the genotypic and seasonal variations in acerola fruit quality, emphasizing the importance of selecting appropriate cultivars and harvest season in maximizing desired quality traits. Breeding programs focused on developing advanced acerola cultivars rich in bioactive compounds should be used, considering the greater influence of genotype compared to seasonal conditions on these compounds.