Nitrogen (N) in soybean (Glycine max L.) plants derived from biological nitrogen fixation was shown to be a sustainable N resource to substitute for N fertilizer. However, the limited water supply in sandy soil is a critical factor for soybean nodulation and crop growth. This study investigated the potential mechanism of the effect of biochar and irrigation on the soybean-Rhizobium symbiotic performance and soil biological activity in a field trial. In the absence of N fertilizer, 10 t ha−1 of black cherry wood-derived biochar were applied under irrigated and rainfed conditions on an experimental, sandy field site. The plant biomass, plant nutrient concentrations, nodule number, nodule leghemoglobin content, soil enzyme activities, and soil-available nutrients were examined. Our results show that biochar application caused a significant increase in the nodule number by 35% in the irrigated condition. Shoot biomass and soil fluorescein diacetate hydrolytic activity were significantly increased by irrigation in comparison to the rainfed condition. The activity of soil protease reduced significantly, by 8%, with the biochar application in the irrigated condition. Further, a linear correlation analysis and redundancy analysis performed on the plant, nodule, and soil variables suggested that the biochar application may affect soybean N uptake in the sandy field. Nodulation was enhanced with biochar addition, however, the plant N concentration and nodule Lb content remained unaffected.