Slave-making (dulotic) ants have long fascinated biologists because of their intriguing behavior and highly specialized lifestyle. Dulosis evolved convergently several times within the two ant subfamilies Myrmicinae and Formicinae. Here, we demonstrate that it originated at least six times independently within the small myrmicine tribe Formicoxenini alone. Our phylogenetic trees, based on 1386 base pairs of the mitochondrial cytochrome oxidase gene, document different degrees of genetic divergence between different monophyla of slave-makers and their host species, which suggests that they evolved from non-parasitic Formicoxenini at different times. Two nearctic slave-makers, Temnothorax duloticus and a new species still to be formally described, appear to be of particularly recent origin. In contrast, the other parasitic monophyla clearly diverged much earlier from their nonparasitic ancestors and have a much longer evolutionary history.

We analysed the phylogenetic and phylogeographic relationships of four Mediterranean species of the rare slave-making ant genus Chalepoxenus and eleven of its about 20 Temnothorax host species by sequencing the mitochondrial Cytochrome Oxidase I and II genes. Neighbour-Joining, Maximum Parsimony and Bayesian analyses based on 1320 bp indicate that the genus Chalepoxenus constitutes a monophylum. In all three analyses, C. kutteri from Southwest Europe and the workerless, "degenerate slavemaker" C. brunneus from North Africa form a monophyletic group. C. muellerianus and C. tauricus, distributed in Southern Europe and Ukraine, respectively, form a monophylum in the Neighbour-Joining and the Maximum Parsimony analysis. In our limited set of only 11 of several hundred Temnothorax species, T. flavicornis forms the sister group of Chalepo-A C H T U N G T R E N N U N G xenus. Our study further indicates paraphyly of the genus Temnothorax with respect to Chalepoxenus. Moreover, the results suggest that speciation in this slave-making genus is possibly caused by the formation of host races as different Chalepoxenus species use different hosts, and some samples seem to cluster by host species rather than by geographical distance.

The remarkable ability of slave-making ants to integrate chemically in the colonies of their host species makes them useful model systems for investigating the role of cuticular hydrocarbons in chemical recognition. The purpose of our study was to examine the influence of the rearing host species on the cuticular hydrocarbon profile and on the mating behaviour of sexuals of the slave-making ant Chalepoxenus muellerianus. Sexuals from a population parasitizing exclusively the host species Temnothorax unifasciatus were reared in the laboratory either with their natural host or another potential host species, Temnothorax recedens. C. muellerianus males reared with T. unifasciatus investigated and mounted female sexuals reared with the same host significantly more often than female sexuals reared with T. recedens. Similarly, C. muellerianus males reared with T. unifasciatus discriminated against female sexuals from natural T. recedens colonies. Males experimentally or naturally reared with T. recedens did not clearly discriminate between female sexuals reared by the two host species and only rarely engaged in mating attempts with either type of female sexuals. Chemical analyses showed that host species affect the chemical profile of C. muellerianus sexuals and vice versa. Our findings indicate that cuticular hydrocarbons might be important in the mating success of this ant species.