Hydrological Processes published by John Wiley & Sons Ltd Perennial cellulosic crops are promoted for their potential contributions to a sustainable energy future. However, a large-scale perennial bioenergy production requires extensive land use changes through diversion of croplands or conversion of uncultivated lands, with potential implications for local and regional hydrology. To assess the impact of such land use conversions on ecosystem water use, we converted three 22 year-old Conservation Reserve Program (CRP) grasslands and three 50+ year-old conventionally tilled corn-soybean crop fields (AGR) to either no-till continuous maize (corn) or perennial (switchgrass or restored prairie) bioenergy crops. We also maintained one CRP grassland without conversion. We measured evapotranspiration (ET) rates on all fields for 9 years using eddy covariance methods. Results show that: (a) mean growing-season ET rates for perennial crops were similar to the ET rate of the corn they replaced at the previously cultivated (AGR) field but ET rates for perennial crops at CRP fields were 5–9% higher than ET rate for corn on former CRP fields; and (b) mean nongrowing season ET rates for perennial fields were 11–15% lower than those for corn fields, regardless of land use history. On an annual basis, mean ET rates for perennial crops tended to be lower (4–7%) than ET rate of the corn that they replaced at AGR fields but ET rates for perennial crops and corn at CRP fields were similar. Over 9 years, mean ET rates for the same crop across land use histories were remarkably similar for corn, whereas for the perennial crops they were 4–10% higher at former CRP than at former AGR fields, mainly due to differences in growing season ET. Over the 9 years and across all fields, ET returned ~60% of the precipitation back to the atmosphere. These findings suggest that large-scale substitution of perennial bioenergy crops for rainfed corn in mesic landscapes would have little if any (0 to −3%) impact on terrestrial water balances.