Kiwifruits are often exposed to various temperature fluctuation (TF) during postharvest storage. To evaluate the effect of TF on the qualities of kiwifruits, fruits were stored at 2 ºC, 2 ºC or 5 ºC (TF2ºC-5ºC), 2 ºC or 7 ºC (TF2ºC-7ºC), alternating every 12 h. Observations revealed that kiwifruits stored at constant 2 °C showed the lowest loss of weight and vitamin C due to the minimum ethylene production and respiratory rate compared to that of TF2ºC-5ºC and TF2ºC-7ºC. Moreover, the results of RT-qPCR verifed that the expression of genes encoding polygalacturonase, β-galacturonidase, and pectin methyl esterase was significantly increased by the treatment of TF. Hence, TF accelerated the degradation of the cell walls, softening, translucency and relative conductivity of the flesh of kiwifruits. Besides, the impact of TF2ºC-7ºC on kiwifruits was more significant relative to TF2ºC-5ºC. The present study provides a theoretical basis for kiwifruit during cold storage.