Microbial contamination is one of the main reasons for the quality deterioration of postharvest blueberries during storage. In this study, we investigated the surface microbiota of blueberry fruits stored at different temperatures via high-throughput sequencing of the 16S and ITS rRNA genes. The results indicated that the α-diversity of the microbial communities in samples stored at 4 °C was much higher than that in samples stored at 25 °C. The composition of the bacterial and fungal communities on the surface of the blueberry fruits varied at different storage temperatures. Ascomycota, Basidiomycota, Anthophyta, Chlorophyta, Proteobacteria, and Cyanobacteria were the most abundant phyla in the bacterial community. Furthermore, five preservation quality indices were measured, and the influence on the α-diversity of the bacterial community was found to be significantly weaker than that of the fungal community. Based on the prediction of the bacterial flora function, the change in blueberry quality during storage was closely related to its surface microbial effect. This study provides a theoretical basis for an understanding of the microbiota on the surface of blueberry fruits to cause fruit spoilage, and the development of a targeted inhibition technology to preserve blueberry fruits under different storage and transportation environments.