BackgroundRiboflavin, a vital water‐soluble vitamin with antioxidative activity, plays a critical role in maintaining overall bodily health and defense responses. However, its impact on fragrant rice yield and aroma remains unexplored.ResultsIn a 2022 pot experiment with Meixiangzhan and Yuxiangyouzhan fragrant rice cultivars, we applied riboflavin foliar treatments at concentrations of 0 (CK), 10 (R10), 20 (R20), and 40 (R40) mg·L‐1 during the initial heading stage. Riboflavin increased rice yield, 2‐AP content, and antioxidative properties. It boosted 2‐AP level by 13.1‐50.1% for Meixiangzhan and 22.3%‐35.3% for Yuxiangyouzhan, with the highest levels in R20 and R10 treatments. This increase is significantly correlated with elevated levels of proline, pyrroline‐5‐carboxylic acid, 1‐pyrroline, and methylglyoxal, as well as heightened enzyme activities, including those of proline dehydrogenase, ornithine aminotransferase, and pyrroline‐5‐carboxylic acid synthetase (P5CS). The R20 treatment resulted in the highest yield due to an improved seed‐setting rate. Importantly, a positive correlation emerged between 2‐AP content and yield, both significantly linked to superoxide dismutase, proline, hydrogen peroxide, P5CS, catalase, and 1‐pyrroline.ConclusionsRiboflavin maintained enzyme activities, regulated substance synthesis pathways, and increased 2‐AP and yield, especially in the R20 treatment. These insights advance fragrant rice production theory by uncovering riboflavin's role in the development of fragrant rice.This article is protected by copyright. All rights reserved.