Sweet cherry is one of the most popular fresh fruits in the world. Previously, we isolated a soilborne dark septate endophyte (DSE) strain, S16, which promoted the growth of Gisela 5 sweet cherry rootstock. However, little is known about the molecular mechanism of the effect of S16 on the growth of sweet cherry. In this study, the physiological parameters and transcript profiles of sweet cherry roots were analyzed under S16 treatment compared with a control to elucidate the molecular mechanisms of the effect of this strain on sweet cherry growth. After inoculation with S16, sweet cherry seedlings exhibited more vigorous growth. Moreover, we identified 4249 differentially expressed genes (DEGs) between S16-treated plants and the control. Many of the DEGs are involved in pathways related to plant growth, such as cellular metabolic and plant hormone pathways. Additionally, some genes involved in nitrate regulation were also enriched; and these genes may be involved in the regulation of nitrate uptake in plants. Physiological index detection demonstrated that S16 could improve the nitrate assimilation of sweet cherry via NO3– transporters. This RNA-seq dataset provides comprehensive insight into the transcriptomic landscape to reveal the molecular mechanisms whereby the DSE influences the growth of sweet cherry.