An assessment of leaf anatomic traits of pecan [Carya illinoinensis (Wangenh.) C. Koch] cultivars (Pawnee, Mohawk, and Starking Hardy Giant) collected from three locations (Tifton, GA; Chetopa, KS; and Stillwater, OK) was conducted to provide an understanding of patterns of ecogeographical variation within the natural range. Acetate casts of representative leaves were prepared for microscopic characterization of epidermal traits (stomatal density, stomatal index, and epidermal cell density). There were differences among the three pecan cultivars at the same location, but there were no differences in stomatal density within the same cultivar grown at three distinct locations. The stomatal density of ‘Pawnee’ leaves (404 stomata/mm2) was intermediate between that of ‘Mohawk’ (363 stomata/mm2) and ‘Starking Hardy Giant’ (463 stomata/mm2). ‘Pawnee’ had the greatest epidermal cell density (2511 cells/mm2) whereas ‘Starking Hardy Giant’ showed the least (1414 cells/mm2). Within a location, stomatal index differed significantly among cultivars, with ‘Starking Hardy Giant’ having a greater stomatal index than the other two cultivars. There were no differences in stomatal index across locations. ‘Mohawk’ had the greatest trichome density (18.92 trichomes/mm2) whereas ‘Starking Hardy Giant’ had the lowest (9.6 trichomes/mm2). The study suggests that differences in stomatal density and epidermal cell density in pecans are cultivar specific rather than being determined by environmental factors. The stability of certain leaf anatomic characteristics, such as stomatal and epidermal cell density, for pecan cultivars grown at different locations confirms that these traits can be used for screening provenances with desirable leaf anatomic characteristics for breeding and cultivar development.