Formica truncorum is an ant species that maintains populations dominated by either single-queen (monogyne) or multiple-queen (polygyne) colonies. New queens (gynes) from monogyne colonies disperse after mating whereas gynes from polygyne colonies are philopatric. Gyne physical condition frequently reflects the overall dispersal propensity of the maternal colony, with dispersers heavier and with larger wings than philopatric morphs. Little, however, is known about the musculature that powers flight dispersal. Our histological examination of alary muscle tissue revealed that mitochondrion and sarcomere numbers did not differ in newly emerged gynes from monogyne and polygyne populations. Significantly greater numbers of mitochondria and sarcomeres were, however, detected in mature virgin gynes from monogyne nests. Within both populations, gynes that had shed their wings had significantly fewer mitochondria than winged gynes. Wingless mated gynes from monogyne nests also had greater numbers of mitochondria. As the sites of ATP production, mitochondria are critical to any biochemical process and reduced numbers are likely to influence the success of flight dispersal. Differential provisioning of new gynes post-emergence in the two populations may account for the variation in flight musculature, which may provide a reliable mechanism by which current dispersal conditions can be assessed. As flight capability generally represents a trade-off with ovarian growth for many insect species, gynes able to forgo flight dispersal and redirect resources from flight muscle development into oocyte development may gain an early reproductive edge compared to their dispersing counterparts.