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The uneven maturity and severe internal rot of kiwifruit were rapidly accelerated when transferred to room temperature after long‐term controlled atmosphere storage. In this study, the effects of ethrel combined with different flow microcirculation of antiseptic mediums on the shelf‐life quality of “Qinmei” kiwifruit stored at 20°C were evaluated. Results indicated that the combined aerosolization application of 500 mg/L natamycin, 7.5% w/w H2O2 and 0.5 mg/L atomized ozone could control the decay incidence and respiratory rate of ethrel‐treated kiwifruit. Further studies indicated that ethrel and O3 treatment greatly increased the activities of chitinase, β‐1,3‐glucanase, catalase, and superoxide dismutase and produced higher ascorbic acid levels (151.16 mg·100 g−1/FW) to enhance disease‐resistant of kiwifruit. This treatment could inhibit the activity of polyphenol oxidase and delay the process of membrane lipid peroxidation, contributing to preserving the integrity of cell structures well. Moreover, ethrel + O3 treatment could effectively reduce ethylene production (3.12 μl/kg/h) to delay the ripening and senescence of fruit stored at 20°C. More importantly, this treatment exhibited the lowest contents of decay incidence (9.75%), total bacterial count (4.07 log cfu/g), and yeasts and molds (4.18 log cfu/g), and thus extended the shelf life for up to 16 days. Therefore, a combination of ethrel and ozone is a potentially effective method for the maintenance of the shelf‐life properties of kiwifruit after long‐term cold storage. Novelty impact statement In this study, the treatment of ethrel‐treated with aerosolized ozone of flow microcirculation of 0.5 mg/L O3 could effectively suppress ripening and senescence, enhance antioxidant capacity and disease resistance, reduce the degree of membrane lipid peroxidation, and considerably preserve the integrity of cell structures, which contributed to preventing the penetration of pathogens, retarding the decay incidence, and delaying the shelf‐life quality deterioration of kiwifruit transferred to room temperature after long‐term controlled atmosphere storage.

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