This study investigated the possible mechanism of softening and senescence of blueberry after harvest using chitosan/thyme oil coating combined with UV-C (short wave ultraviolet irradiation) treatment. On the 56th day of storage, the CBP, cellulose, and hemicellulose contents in the chitosan/thyme oil coating +UV-C-treated group were 1.41, 1.65, and 1.20 times higher than those in the control group. Compared with the control group, the activities of polygalacturonase (PG), pectin methylesterase (PME), β-glucosidase (β-Gal), and cellulose (Cx) were significantly reduced (p < 0.05) after chitosan/thyme oil coating +UV-C, and their maximum values decreased by 5.41 μg/h g, 5.40 U/g, 12.41 U/g, and 3.85 μg/h g, respectively. Moreover, chitosan/thyme oil coating combined with UV-C treatment inhibited the gene expression of PG, PME, Cx, and β-Gal and then regulated the decrease in PG, PME, Cx, and β-Gal activities, inhibited the degradation of cell wall polysaccharides, and delayed the softening and senescence of postharvest blueberries. The results showed that chitosan/thyme oil coating, UV-C, and chitosan/thyme oil coating + UV-C could significantly inhibit postharvest softening of blueberry; chitosan/thyme oil coating +UV-C had the best effect.