Populations of American beech in Virginia and the Great Smoky Mountain National Park in Tennessee and North Carolina were investigated for demographic genetic substructurings. Two Virginia populations, one on the Blue Ridge (WG1) and the other on the Piedmont (WG2) occur over an elevational gradient of several hundreds meters. One of the Great Smoky Mountain populations (GS1) was in a ‘beech gap’ and the other (GS2) in a ‘cove forest’ along a creek. The populations in the Great Smoky Mountain National Park were only separated by a few hundred meters in elevation, but both on the same physiographic province. The populations had two growth forms. Trees produced extensive root suckers at WG1, GS1 and GS2, but WG2 had no root suckers and all individuals had obviously been established from seeds. A total of 1335 shoots were mapped at the four sites, their size measured [diameter at breast height (DBH) or diameter at ground height (DGH)], and genotypes were determined for each locus using allozyme analysis. FIS among five different size-classes revealed an excess of homozygotes in WG1, GS1 and GS2, and an excess of heterozygotes in WG2. The offshoot formation from root suckers obviously contributed to the abundance of intermediate size-classes in WG1, GS1 and GS2. Exceedingly localized patchiness of different multilocus genotypes reveals genetic clustering of shoots that have obviously originated from root suckers in WG1, GS1 and GS2. The Piedmont population (WG2), on the other hand, showed loose localization of genetically related trees at a scale of 35–40 m in area, suggesting broader ranges of pollen and seed dispersal. The data are discussed in the light of the differences in growth form and mode of reproduction, and also in relation to the post-glacial migration and the current geographic distribution of the species.