Fresh peaches and nectarines are very popular for their high nutritional and therapeutic value. Unfortunately, they are prone to rapid deterioration after harvest, especially if the cold chain is not well maintained. The objective of this work is to study the environmental fluctuation and the quality change of fresh peaches and nectarines in cold chain. The temperature, relative humidity, and CO2 level were real-time monitored by sensor nodes with a wireless sensor network (WSN). The cold chain lasted for 16.8 h and consisted of six segments. The dynamic change of temperature, relative humidity, and CO2 level were real-time monitored and analyzed in detail in each of the six stages. The fruit quality index (fruit weight, fruit firmness, and soluble solids concentration (SSC)) were detected and analyzed immediately before the first stage (S1) and at the beginning of the last stage (S6). The results show that without good temperature control fruit softening is the most significant problem, even in a short chain; the WSN node can provide complete and accurate temperature, humidity, and gas monitoring information for cold chains, and can be used to further improve quality and safety assurance for peach fruit cold chains.