Application of hydrogen-rich water maintains red pitaya fruit quality through regulation of ROS and energy metabolism

Hydrogen-rich water (HRW) treatment has been shown to enhance the shelf life of fruit and vegetables. This study aimed to investigate the effects of HRW on improving the quality of red pitaya and its underlying regulatory mechanisms. The results revealed that applying HRW at a 70% concentration reduced decay and weight loss rates, slowed the respiratory rate, and delayed the decline in titratable acidity and total soluble solids. Additionally, HRW treatment mitigated the increase in superoxide anion production and hydrogen peroxide levels, while enhancing antioxidant enzyme activities and gene expression, including those of peroxidase, superoxide dismutase, glutathione peroxidase, catalase, ascorbate peroxidase, monodehydroascorbate reductase, and glutathione reductase. Non-enzymatic antioxidant compounds such as ascorbate and glutathione also increased. Moreover, RT-qPCR analysis showed that HRW treatment upregulated HcMYB6 and downregulated HcR2R3MYB. Dual-luciferase reporter assays indicated that HcMYB6 activated the expression of HcSOD and HcCAT, whereas HcR2R3MYB repressed their expression. These findings suggest that HcMYB6 and HcR2R3MYB may be involved in reactive oxygen species metabolism, with HcMYB6 being induced and HcR2R3MYB inhibited by HRW treatment. Furthermore, HRW treatment led to increased adenosine triphosphate and adenosine diphosphate levels, a decrease in adenosine monophosphate content, and elevated gene expression and activity of Ca²⁺-ATPase, succinate hydrogenase, H⁺-ATPase, and cytochrome C. Collectively, these results demonstrate that HRW treatment effectively reduces oxidative damage and improves energy status, thereby preserving the quality of red pitaya fruit.