The thrifty genotype hypothesis initiated speculation that feast and famine cycling throughout history may have led to group-specific alterations of the human genome, thereby augmenting the capacity for excessive fat mass accrual when immersed in the modern-day obesogenic environment. Contemporary work, however, suggests alternative mechanisms influencing fuel utilization and subsequent tissue partitioning to be more relevant in the etiology of population-based variation in adipose storage. The objective of this study was to evaluate the independent and interactive contribution of ancestral admixture as a proxy for population-based genetic variation and diet on adipose tissue deposition and distribution in peripubertal children and to identify differences in racial/ethnic and sex groups. Two-hundred seventy-eight children (53% male) aged 7-12 years, categorized by parental self-report as African- (n = 91), European- (n = 110), or Hispanic American (n = 77), participated. Ancestral genetic admixture was estimated using 140 ancestry informative markers. Body composition was evaluated by dual-energy X-ray absorptiometry; energy expenditure by indirect calorimetry and accelerometry; and diet by 24-h-recall. Admixture independently contributed to all adiposity parameters; i.e., estimates of European and Amerindian ancestries were positively associated with all adiposity parameters, whereas African genetic admixture was inversely associated with adiposity. In boys, energy intake was associated with adiposity, irrespective of macronutrient profile, whereas in girls, the relationship was mediated by carbohydrate. We also observed moderating effects of energy balance/fuel utilization of the interaction between ancestral genetic admixture and diet. Interactive effects of genetic and non-genetic factors alter metabolic pathways and underlie some of the present population-based differences in fat storage. © 2011 Wiley-Liss, Inc.