Reduced tillage systems have become more common than conventionally tilled systems in much of the northern Great Plains, raising questions on potential differences in nutrient distribution and availability between tillage systems. Specifically, vertical stratification of phosphorus (P) has been documented in both no-till and minimum till systems, yet very few studies have determined if this stratification has affected P uptake. In this study, stratification of P was compared in 3 cm layers in a small plot study composed of four tillage systems: long-term conventional (sweep) till (T), 10-yr no-till (NT), 1-yr NT and 1-yr T. Olsen P was measured in each layer in the upper 30 cm, and a sequential extraction was performed on all layers in the upper 15 cm. Fractions measured in this extraction included resin-P, NaHCO3-P (bic-P), NaOH-P, and HCl-P. Olsen P concentrations in all tillage systems were approximately 6 fold higher in the surface 3 cm than below 12 cm, demonstrating a substantial amount of vertical P stratification. There were similar trends in concentrations of resin-P, bic-P and NaOH-P in the upper 15 cm. There were very few significant differences in the concentrations of P fractions among tillage systems, possibly because there was only one tillage pass per year. Aboveground uptake of P by a winter wheat crop in 2006 did not differ significantly between tillage systems. The best correlation between wheat P uptake and available P concentrations (Olsen P and resin-P) occurred in the 6 to 9 cm depth, suggesting that this may be the best depth to band fertilizer P. Based on this study, combined with other recent studies that found similar results, fertilizer P rates for small grains likely do not need to be adjusted for tillage system.