Sulfur is an indispensable nutrient for plant growth and development, and is important in the synthesis of sulfurcontaining amino acids. Although several studies on the effects of some macronutrients, including nitrogen and phosphorus, have been conducted on the performance of several crops at the genomic level, studies on the effect of sulfur on crop performance are limited. Therefore, this study aimed to examine the effect of different sulfur concentration on the transcriptome of soybean. Additionally, soybean yield parameters were also examined. Two soybean varieties, DND252 and HN84, were exposed to low and high concentrations of sulfur, and differentially expressed genes (DEGs) were identified using transcriptome analysis. The study results showed that the DEGs identified in the DND252 variety were involved in stimuli response, DNA binding and cell periphery under low sulfur concentrations. Also, the DEGs identified under high sulfur concentration were involved in membrane and membrane parts. Additionally, DEGs identified in the HN84 variety under low sulfur concentrations had similar functions as those identified in DND252 under high sulfur concentrations, indicating that HN84 was more sensitive to sulfur concentration changes than DND252. However, under higher sulfur concentrations, the DEGs identified in HN84 were primarily involved in membrane and membrane parts, indicating that high sulfur can cause cell membrane damage. Furthermore, soybean grown using 2.0 mmol/L sulfur had the best yield. The findings of this study identified candidate genes for the breeding and development of sulfur-efficient soybean varieties.