Sex and recombination are central processes in life generating genetic diversity. Organisms that rely on asexual propagation risk extinction due to the loss of genetic diversity and the inability to adapt to changing environmental conditions. The fungus-growing ant species Mycocepurus smithii was thought to be obligately asexual because only parthenogenetic populations have been collected from widely separated geographic localities. Nonetheless, M. smithii is ecologically successful, with the most extensive distribution and the highest population densities of any fungus-growing ant. Here we report that M. smithii actually consists of a mosaic of asexual and sexual populations that are nonrandomly distributed geographically. The sexual populations cluster along the Rio Amazonas and the Rio Negro and appear to be the source of independently evolved and widely distributed asexual lineages, or clones. Either apomixis or automixis with central fusion and low recombination rates is inferred to be the cytogenetic mechanism underlying parthenogenesis in M. smithii. Males appear to be entirely absent from asexual populations, but their existence in sexual populations is indicated by the presence of sperm in the reproductive tracts of queens. A phylogenetic analysis of the genus suggests that M. smithii is monophyletic, rendering a hybrid origin of asexuality unlikely. Instead, a mitochondrial phylogeny of sexual and asexual populations suggests multiple independent origins of asexual reproduction, and a divergence-dating analysis indicates that M. smithii evolved 0.5-1.65 million years ago. Understanding the evolutionary origin and maintenance of asexual reproduction in this species contributes to a general understanding of the adaptive significance of sex.Attini | clonality | Formicidae | thelytoky | mutualism T he vast majority of metazoans reproduces sexually, enjoying the benefits of genetic recombination (1-3) such as rapid adaptability to novel ecological conditions (4, 5) and the purging of deleterious mutations from their genomes (6, 7). However, relative to sexually reproducing organisms, an asexual female doubles its fitness by transmitting its entire genetic material to the next generation (8). Despite such obvious short-term fitness advantages, asexual organisms occur only sporadically throughout the tree of life and are predicted to be evolutionarily short-lived and doomed to early extinction (9-11). In contrast to the short-term advantages of asexuality, the adaptive value of sexuality, that is, genetic recombination, is expected to be of long-term benefit (2, 12-14). There remain in evolutionary biology significant unexplored questions about whether sexual reproduction is favored by natural selection over short evolutionary time spans and, if not, why sexual reproduction persists as the prevalent mode of reproduction, given that the selective benefits are deferred. Studying the origin and evolution of parthenogenetic lineages, and understanding how genetic diversity is generated and preserved in such linea...