Alabama ranks second in broiler production nationwide generating 1.5 million tons of poultry litter (PL) each year. Many row crop farmers in Alabama are using PL as a slow-release nutrient source in their row crop production system including soybean (Glycine max L.). However, little is known about the effect of PL on soybean grain yield and nutrient composition. The objective of this study was to evaluate the effect of PL on soybean grain yield and seed nutrient composition in response to three application rates (1, 2.5, and 5 tons/acre). A two-year replicated field plot study was conducted during the 2018 and 2019 growing season on a Compass loamy sand in central Alabama. Treatments included pre-plant PL application (2 weeks before planting) at three rates (1, 2.5, and 5 tons/acre) and a control plot (no litter). Soybean cultivar (AG74X8; Maturity group-7) was planted on June 14^th and harvested on 20^th November. Plot yields were determined by combine harvesting the middle two rows. Grain nutrient content of total nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), calcium (Ca), sulfur (S), boron (B), zinc (Zn), manganese (Mn), iron (Fe) and copper (Cu) were determined according to the routine procedure. Data analyses were performed using PROC GLIMMIX of SAS 9.4. Poultry litter significantly affected soybean grain yield in 2019 (P<0.05). The PL-treated plots had a 42% greater grain yield than the control plots. The PL application rate of 5 tons/acre resulted in the highest grain K and Cu concentrations and differed significantly (P<0.05) from the 2.5 ton/acre application rate and the control treatment.