Climate, specifically moisture and temperature, influences plant growth and nutrient cycling; thus, climate also influences prairie development. Relatively little research has been conducted in the ecological transition zone between the Great Plains and the more humid forests. Since moisture and temperature affect many ecosystem processes, the wetter climate of the southern forest region should influence soil biogeochemical cycling quite differently from that in grassland soils. The objective of this study was to evaluate soil quality and the relationships between selected soil properties across a climosequence in a transition zone between tallgrass prairie in humid-temperate (Ozark Highlands) and humid-subtropical (Grand Prairie) climate regimes in Arkansas. Soil physical, chemical and biological properties of the upper 10 cm differed significantly between physiographic regions. Linear relationships between total soil nitrogen (N) and carbon (C), extractable phosphorus and manganese, electrical conductivity and soil organic matter concentration differed significantly between physiographic regions. Total soil N and C decreased with increasing soil bulk density in both physiographic regions. The relationship between total C and bulk density differed by physiographic region, while the relationship between total N and bulk density did not. Soil organic matter concentration, C:N ratio, and many extractable nutrients, were higher and relationships between selected soil properties differed under native tallgrass prairie in a relatively warm and wet climate than that in a relatively cooler and drier climate. The results of this study suggest that prairie preservation, restoration and management practices should differ depending on climate regimes.