Red light irradiation can effectively prolong the shelf-life of many fruit. However, little is known about red light-induced metabolite and enzyme activities. In this study, pitaya fruit was treated with 100 Lux red light for 24 h. Red light irradiation significantly attenuated the variation trend of senescence traits, such as the decrease of total soluble solid (TSS) and TSS/acidity (titratable acidity, TA) ratio, the increase of TA, and respiratory rate. In addition, the reactive oxygen species (ROS) related characters, primary metabolites profiling, and volatile compounds profiling were determined. A total of 71 primary metabolites and 67 volatile compounds were detected and successfully identified by using gas chromatography mass spectrometry (GC-MS). Red light irradiation enhanced glycolysis, tricarboxylic acid (TCA) cycle, aldehydes metabolism, and antioxidant enzymes activities at early stage of postharvest storage, leading to the reduction of H2O2, soluble sugars, organic acids, and C-6 and C-7 aldehydes. At a later stage of postharvest storage, a larger number of resistance-related metabolites and enzyme activities were induced in red light-treated pitaya peel, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging, reducing power, fatty acids, and volatile aroma.