Seasonal patterns in xylem pressure potential (psi) and stomatal conductance to water vapor (g) were compared between seedlings (1-5 years old) invading tallgrass prairie and adult trees of Quercus macrocarpa (Bur oak) and Quercus muehlenbergii (Chinquapin oak) in adjacent gallery forest. Water is often limiting to productivity in the Flint Hills of NE Kansas, thus, we hypothesized that seedlings invading a grassland would have lower psi and g compared with adult trees, reflecting competitive interactions for water between seedlings and the dominant grasses. We found that psi was usually lower in adult trees than in older seedlings, but in the very smallest seedlings psi was significantly reduced compared with mature (greater than 50 years) individuals. Adult and 1- to 2-year-old seedlings had similar g during the season with highest g occurring in older (4- to 5-year-old) seedlings. In general, Q. muehlenbergii had higher psi and lower g than Q. macrocarpa consistent with reports that Q. muehlenbergii is more drought tolerant. Rapid and extensive root growth characteristic of Quercus seedlings reduced the period of time when seedlings invading prairie must compete with the grasses for water, although water stress may still be an important factor affecting first-year seedlings in this ecosystem. In contrast, strong intraspecific competition for water within mature gallery forests may have reduced psi and g in forest trees. Thus, the potential for water stress to impact Quercus survival and growth in this drought-prone ecosystem is greatest during establishment and then after forest development is complete. Older seedlings (greater than 4-5 years old), saplings and isolated trees within the prairie may experience the least water stress due to a reduction in both inter- and intraspecific competition for water. These data suggest that in the absence of severe fire or drought occurring during the growing season, gallery forests will continue to expand into tallgrass prairie.