Land‐use change can have a major impact on soil properties, leading to long‐term changes in soilnutrient cycling rates and carbon storage. While a substantial amount of research has been conducted onland‐use change in tropical regions, empirical evidence of long‐term conversion of forested land toagricultural land in North America is lacking. Pervasive deforestation for the sake of agriculturethroughout much of North America is likely to have modified soil properties, with implications for theglobal climate. Here, we examined the response of physical, chemical and biological soil properties toconversion of forest to agricultural land (100 years ago) on Roebuck Farm near Perth, Ontario, Canada.Soil samples were collected at three sites from under forest and agricultural vegetative cover on bothhigh‐ and low‐lying topographic positions (12 locations in total; soil profile sampled to a depth of 40cm).Our results revealed that bulk density, pH, and nitrate concentrations were all higher in soils collectedfrom cultivate sites. In contrast, samples from forested sites exhibited greater water‐holding capacity,porosity, organic matter content, ammonia concentrations and cation exchange capacity. Many of these characteristics are linked to greater organic matter abundance and diversity in soils under forestvegetation as compared with agricultural soils. Microbial activity and Q10 values were also higher in theforest soils. While soil properties in the forest were fairly similar across topographic gradients, low‐lyingpositions under agricultural regions had higher bulk density and organic matter content than upslopepositions, suggesting significant movement of material along topographic gradients. Differences in soilproperties are attributed largely to increased compaction and loss of organic matter inputs in theagricultural system. Our results suggest that the conversion of forested land cover to agriculture landcover reduces soil quality and carbon storage, alters long‐term site productivity, and contributes toincreased atmospheric carbon dioxide concentrations.