Ascorbic acid (AsA) is an indicator of the nutritional value of freshly cut kiwifruit during storage at 4℃, and its degradation can be inhibited after ozone treatment (1 mg/L, 10 min). The aim of this study was to elucidate the regulatory mechanism affecting AsA metabolism in fresh-cut kiwifruit after ozone treatment. In this study, ozone treatment not only prevented the decrease in AsA/dehydroascorbic acid and delayed the accumulation of total soluble solids/titratable acidity, but also altered phytohormone levels differently. Transcriptomic profiling combined with cis-acting element and correlation analysis were performed to reveal that abscisic acid and salicylic acid synergistically delay AsA degradation under ozone-treatment conditions. Actinidia03760, encoding ascorbate peroxidase, could be specifically recognized by the bZIP transcription factor and is considered a key candidate gene for further research. Collectively, ozone treatment is a promising method for preserving AsA content and improving the nutrition of fresh-cut kiwifruit.