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Mango (Mangifera indica) is a highly valuable horticultural crop known for its quality and productivity. This study investigates the dynamic changes in physicochemical properties and glutathione and ascorbic acid metabolic pathways in mango pulp across various growth and development stages over two consecutive years (2021–2022 and 2022–2023) by transcriptomic analysis. Overall, the results demonstrate that during different ripening periods, the pulp shows increased levels of total soluble solids, relative conductivity, glutathione, and enzymes, while titratable acidity, malondialdehyde, reactive oxygen species, and ascorbic acid contents decreased. Moreover, transcriptomic analysis identified key differentially expressed genes from the glutathione and ascorbic acid metabolic pathways and validated them with qRT-PCR. In different comparisons, a total of 1776, 2513, and 828 DEGs were identified in 30 vs. 60, 30 vs. 90, and 60 vs. 90 days after flowering, respectively. Among them, seven DEGs were primarily enriched in relevant pathways, which included ascorbate peroxidase, ascorbate oxidase, glutathione peroxidase, gamma-glutamyl transferase, glutathione transferases, and glucose-6-phosphate dehydrogenase. The upregulation of these genes indicates that glutathione and AsA respond well to scavenging reactive oxygen species and maintain normal functioning in plants. This research sheds light on the molecular mechanisms of glutathione and ascorbic acid dynamic changes in mango pulp, providing valuable insights into the regulation of antioxidant and metabolic pathways during fruit growth and development. [ABSTRACT FROM AUTHOR]