Concerns about global pollinator declines have placed a growing focus on understanding the impact of agriculture practices on valuable native pollinators in these systems. Cultivation practices such as tillage disturb agroecosystems and can have negative impacts on ground-nesting pollinators. The squash bee, Peponapis pruinosa (Say), is a ground-nesting specialist pollinator of Cucurbita (Cucurbitaceae) crops (i.e., pumpkins and squash) that often nests in agricultural fields and thus may be vulnerable to these practices. We investigated the impact of tillage on nesting behavior of P. pruinosa in plasticulture and strip-tilled squash systems. We used choice experiments to test nesting substrate preference and nesting success of caged P. pruinosa in two soil tillage systems: strip tillage and plasticulture. The strip tillage system comprised two tillage zones (strip-tilled row with no-till edges), and the plasticulture system comprised two tillage zones (plastic bed and conventional tillage edge). The results of our study indicate that P. pruinosa nesting density did not significantly differ between the strip tillage and plasticulture systems. Within each system, P. pruinosa preferred excavating nests in the most disturbed soil zones (strip-tilled row and conventionally tilled edge). In the strip tillage system, the strip-tilled row had significantly more nests than the no-till edge. Results of these studies suggest that soil tillage practices can influence P. pruinosa nesting choice and production practices should be considered when developing a pollinator protection plan.