In Ohio and elsewhere, recent grassland plantings in the federal Conservation Reserve Program (CRP) have become much more extensive than native prairie remnants. The seed source for CRP grasslands in Ohio often comes from as far away as Missouri or Texas, which may be undesirable from the standpoint of conservation genetics. The goal of this study was to examine the potential for gene flow from large, recently introduced populations of Big bluestem (Andropogon gerardii, Poaceae) to small local populations of this outcrossing perennial species. We examined the potential for cross-pollination between three local populations and three introduced CRP populations by comparing flowering phenologies. Flowering times overlapped extensively, indicating that cross-pollination is possible where local and introduced genotypes co-occur. To compare genetic variation in local and CRP populations, we analyzed variation at 68 RAPD loci in six populations of each type. Somewhat surprisingly, we found no significant differences in the genetic diversity or composition between the two groups (local vs. CRP). In summary, we found that local and introduced populations of Big bluestem have the potential to interbreed, based on their flowering periods, but further research is needed to determine whether local genotypes harbor unique genetic variation that could be jeopardized by hybridization with introduced genotypes.