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This study of genetic diversity in a wild ancestor of the cultivated strawberry was undertaken to describe patterns of variation in nature, assess worth of existing germplasm collections, and identify promising locations for future collection. Previous work reported a similar study of octaploid strawberry ranging east to west across North America. This complementary study focused on variation from north to south in the Rocky Mountains. The morphological diversity of 16 populations of Fragaria virgininia were characterized for morphological and molecular traits. Two clones of each of 133 genotypes from these populations were grown in a common environment in a greenhouse. Eighteen morphological traits, such as leaf area, runner color, and days to flowering, were measured and analyzed with principal components and canonical discriminant analyses. Molecular diversity data were obtained using seven randomly amplified polymorphic DNA primers. Resulting population marker frequencies were also subjected the previously describe anlayses. Differences due to latitude, longitude, and altitude were observed. Implications of the results will be discussed.