Identification of Key Enzymes and Genes Modulating L-Ascorbic Acid Metabolism During Fruit Development of by Integrating Metabolome, Transcriptome, and Physiological Analysis

is acknowledged for its substantial nutritional benefits, particularly attributed to the high levels of ascorbic acid (AsA) found in its fruits. The "Mengqi No.1" variety of , which is cultivated in Qinghai, is known for its high yield and exceptional quality. We utilized the "Mengqi No.1" variety as experimental materials and combined metabolomic, transcriptomic, and physiological analyses to investigate the metabolites, genes, and enzymes related to AsA metabolism in fruits. The results revealed nine differential metabolites associated with AsA metabolism in fruits across three stages, including 1D-Myo-Inositol-1,4-Bisphosphate, D-Fructose, L-(+)-Arabinose, I-Inositol, L-Arabinitol, D-Galactose-1-P, lactose, α-D-Glucose, and D-Glucose-6-P. Notably, the contents of D-Glucose-6-P, D-Galactose-1-P, and D-Fructose were increased as the fruit developed. Additionally, fresh weight, longitudinal length, and radial width were increased, while the contents of AsA and DHA were decreased. GalDH and DHAR are critical enzymes for the accumulation of AsA and DHA, exhibiting positive correlation coefficient. Furthermore, , , , and were identified as key regulatory genes in the L-Galactose pathway of AsA synthesis, influencing D-Galactose-1-P, D-Glucose-6-P, α-D-Glucose, and D-Fructose. and are considered key positive regulator genes of AsA and DHA in the AsA-GSH cycle. However, the majority of genes (nine) act as negative regulators of AsA and DHA. These findings provide a foundation for the understanding of the regulatory mechanism of AsA metabolism in fruits and offer insights into the utilization of AsA from .