Temperature is a primary determinant of insect and other ectotherm distribution and activity. Physiological and behavioral adaptations allow many insects to survive at subzero temperatures, yet the evolutionary influences on insect cold tolerance are unclear. Supercooling points, basal cold tolerance, cold-tolerance strategy, and inducible cold tolerance from rapid cold-hardening or acclimation were measured in a phylogenetically independent context in larvae of 27 phylogenetically diverse Drosophila species acquired from stock collections. Supercooling capacity is attributed primarily to physical factors, such as dry mass and water mass. Species of the obscura group were more resistant to acute cold tolerance than species of other groups within the genus, and plasticity in cold tolerance is constrained by phylogeny rather than by basal cold tolerance. The more cold-tolerant freeze-avoiding species appear to have arisen multiple times in Drosophila and are distinct from chill-susceptible species, which likely indicate the ancestral state. A phylogenetic influence is apparent on several measures of cold tolerance, which show considerable interspecific variation and indicate varying physiological mechanisms among Drosophila species when temperature limits are met.