In recent years we have investigated the evolution of the Holarctic leaf-beetle genus Timarcha using molecular approaches, but to date several important questions remained unanswered, including its systematic arrangement in a temporal context, or the phylogenetic placement of the Nearctic taxa. Here I present a reanalysis of available genetic data together with newly generated data for key taxa (markers 16S rDNA, CO2, ITS-2, and 18S rDNA), including the Nearctic species (subgenus Americanotimarcha), using direct optimization-based phylogenetic reconstructions. Lineage ages are estimated using maximum likelihood branch-length estimates and the molecular clock calibration derived from several presumed vicariance events in the Mediterranean. Phylogenetic analyses and 18S rDNA divergences suggest the ancient divergence of the Nearctic and Palaearctic lineages, related to the North Atlantic opening in the middle Eocene. The diversification of the Palaearctic Timarcha seems closely related to the geological evolution of the Mediterranean area during the Tertiary, with Pleistocenic climate changes affecting species ranges and lineage extinction, but not resulting in extensive speciation.
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