Fine fescues (Festuca spp.) are cool-season grasses used in low-maintenance turf areas. Mesotrione is a PRE and early-POST herbicide used during establishment of most cool-season turfgrasses, excluding fine fescues. Currently, efforts are being made to breed for increased tolerance to mesotrione in fine fescues to enhance weed control during establishment. This study was conducted to evaluate the association of foliar and root uptake of [14C]mesotrione with the tolerance of three lines each of Chewings fescue [Festuca rubra ssp. commutata Gaudin; syn. F. rubra ssp. fallax (Thuill.) Nyman], hard fescue [Festuca trachyphylla (Hack.) Hack.], and strong creeping red fescue (Festuca rubra L. ssp. rubra) lines. From a rate-titration experiment, the hierarchical rank of species for mesotrione tolerance from highest to lowest was: hard > Chewings > strong creeping red fescue. The hierarchical rank of species for foliar uptake from highest to lowest was: Chewings > strong creeping red > hard fescue. Translocation of foliar-absorbed 14C was not associated with differential tolerance levels of the three species. Root absorption was comparable among species, but differences between lines were detected within the species. The most susceptible lines of Chewings and strong creeping red fescue exhibited greater root uptake than lines with greater tolerance. Hard fescue translocated the least amount of root-absorbed radioactivity to shoots, while Chewings and strong creeping red fescues were comparable.