Estimating hybridization as a consequence of climatic fluctuations: A case study of two geometridae species.

In some cases, the phylogenetic analysis based on the mitochondrial DNA (mtDNA) and the nuclear DNA (ncDNA) are discordant. There are three major causes of the discordance within insects, including hybridization, incomplete lineage sorting (ILS) and infection by Wolbachia. In this study, we used a combination of multilocus and coalescent analyses to explore these processes occurred during the evolutionary history of Limbatochlamys rosthorni Rothschild, 1894 and Limbatochlamys pararosthorni Han and Xue, 2005. The ncDNA phylogenetic tree supported two reciprocally monophyletic species, whereas the mtDNA results failed to reveal such a structure and revealed an extensive level of admixture between two species. Because of very low Wolbachia infection rates (<20%), we firstly excluded this reason for the discordance. The fixed nucleotide differences and large genetic distances (1.5-2.5%) at the ncDNA genes suggested that the lineage sorting process between these two species is nearly complete and two species have experienced a prolonged period of independent evolution. Thus, we secondly excluded ILS. Sharing haplotypes, mtDNA gene flow occurring and the transitional samples with morphological features supported hybridization. The distribution contraction during glaciations and postglacial distribution expansion might have facilitated hybridization. Taken together, our study indicates that the current genetic structure of L. rosthorni and L. pararosthorni is the results of contraction and fragmentation into separated refugia during glaciations, followed by postglacial expansion and admixture.