Scholars have proposed the practice of split N fertilizer application (SNFA), which has proven to be an effective approach for enhancing N use efficiency. However, the effect of SNFA on NH3 volatilization, nitrification and denitration in soil, remain largely unknown. As such, the current study assessed soil NH3 volatilization, nitrification and denitrification intensities, abundance of nitrogen cycle-related funetional genes, and invertase activity for different treatments. We applied a rate of 240 kg·ha-1 of N, and the following fertilizer ratios of the percent base to that of topdressing under water-saving irrigation: N1 (basal/dressing, 100%/0%), N2 (basal/dressing, 70%/30%), N3 (basal/dressing, 50%/50%), N4 (basal/dressing, 30%/70%), and N5 (basal/dressing, 0%/100%). N3 treatment resulted in a significant decrease in rate of NH3 volatilization. This treatment also significantly reduced nitrification and denitrification intensities, primarily owing to the reduced functional genes abundance involved in the nitrogen cycle (Amoa-AOB, nirK and nirS) and reduced invertase activity (urease, nitrate reductase, nitrite reductase) in wheat-land soil. 15N tracer studies further demonstrated that N3 treatments significantly increased the grain nitrogen accumulation by 9.50-28.27% compared with that under other treatments. This increase was primarily due to an increase in the amount of N absorbed by wheat from soil and fertilizers, which was caused by an enhancement in total N uptake (7.2-21.81%). Collectively, these results suggest that the N3 treatment (basal/dressing, 50%/50%) improves N uptake by wheat, reduces the soil NH3 volatilization rate, and has the potential to reduce the amount of N2O generated by nitrification and denitrification.