Assortative mating between two sympatric closely-related specialists: inferred from molecular phylogenetic analysis and behavioral data.

Host plant shifting of phytophagous insects can lead to the formation of host associated differentiation and ultimately speciation. In some cases, host plant specificity alone acts as a nearly complete pre-mating isolating barrier among insect populations. We here test whether effective pre-mating isolation and host-independent behavioral isolation have evolved under the condition of extreme host specilization using two sympatric flea beetles with incomplete post-mating isolation under laboratory conditions. Phylogenetic analysis and coalescent simulation results showed that there is a limited interspecific gene flow, indicating effctive isolation between these species. Three types of mating tests in the absence of host plant cues showed that strong host-independent behavioral isolation has evolved between them. We conclude that almost perfect assortative mating between these two extreme host specialists results from a combination of reduced encounter rates due to differential host preference and strong sexual isolation.