Intermediate wheatgrass (Thinopyrum intermedium; IWG) is a perennial grass under development as a grain and forage crop. Although IWG is known for its ability to take up nitrate and improve water quality, seasonal nitrogen (N) demand and uptake by IWG is not well known. We measured IWG shoot, root, and grain production, tissue N concentration, and soil mineral N at multiple plant growth stages in 1- and 2-year-old IWG stands fertilized with various rates of N: (1) 80 kg N ha−1 applied at spring regrowth (spring), (2) 40–40 kg N ha−1 applied at spring regrowth and anthesis (split), and (3) an unfertilized control. We also calculated nitrogen use efficiency and biomass N yield. Soil mineral N, N-mineralization rates, and plant N concentration increased with fertilization, and lodging increased with spring fertilization, while root physiological N use efficiency (PNUE) declined with fertilization. Seasonally, shoot and root N concentration declined at physiological maturity, while shoot PNUE was highest at maturity, suggesting either that surplus N was allocated to grain or that more biomass was being produced per unit N taken up. In the 1-year-old stand, during fall regrowth, soil mineral N levels were among the lowest; however, the total soil N was highest compared with other sampling times, suggesting a large influx of organic N between physiological maturity and fall regrowth. Based on our results, IWG is well suited to use nitrogen inputs and avoid excess N leaching into groundwater, but it is also clear that IWG has strong seasonal N allocation patterns that should be taken into consideration with N recommendations and best practices.