Abstract
Sugar content critically determines mango fruit quality and varies significantly among varieties. Preliminary studies indicate that fructokinases (MiFRKs) MiFRK1 and MiFRK2 likely regulate intervarietal sugar variation. We characterized these MiFRKs using heterologous expression in tomato. Both isoforms phosphorylate fructose, promoting downstream catabolism, with R-MiFRK2 (from low-sugar 'Renong No. 1') exhibiting higher activity than T-MiFRK2 (high-sugar 'Tainong No. 1') and MiFRK1. Transcriptomic and metabolic analyses reveal that MiFRK overexpression inhibits sugar accumulation by altering the expression of key metabolic genes, including sucrose degradation enzymes (invertases), starch breakdown genes (β-amylases), and glycolytic genes (enolases). Intriguingly, MiFRK1 and MiFRK2 exhibit distinct regulatory effects on these pathways, suggesting functional specialization between the two isoforms. These findings provide novel insights into the molecular mechanisms through which MiFRKs govern sugar metabolism in mango, highlighting their potential as key targets for metabolic engineering to enhance fruit quality.