A new mountain lizard from Montes de León (NW Iberian Peninsula): Iberolacerta monticola astur ssp. nov. (Squamata: Lacertidae).
Iberolacerta populations from the Northern Montes de Leon (NML) were studied by means of external morphology (scalation and biometry), osteology and genetics (mtDNA and microsatellites), searching for their homogeneity ("intrazonalanalysis") and, once verified, comparing them with Iberolacerta monticola s. str. (from Central Cantabrian Mountains)and/. gal ani (from Southern Montes de Leon) ("extrazonal analysis") from neighboring areas.Our "intrazonal analysis" revealed discordances between the different approaches, especially the patterns of variation of nuclear microsatellites (congruent with external morphology) and mtDNA, namely a very low nuclear differentiation between relatively highly differentiated mtDNA lineages. The morphological approach was unable to discriminate any of the populations as significantly different from the others in the NML. Mitochondrial DNA revealed a haplotype lineage closely related to I. galani (MNL-II in our text) in some specimens of Sierra de Villabandfn and Suspiron, but these populations are morphologically indistinguishable from the main part of the other populations that belong to lineage NML-1,phylogenetically closer to/. monticola. After a separation from I. manti cola ca. 1.8 Mya, the populations in this geographic region must have suffered at least two different waves of gene flow from I. gal ani, the second one not much later than 0.5 Mya. Microsatellite results indicate that all the NML populations are genetically similar in terms of their nuclear genomes,independently of their mitochondrial differentiation (NML-I vs. NML-II haplotype groups). Since all the morphological and microsatellite evidences point towards the fact that, independently of the mitochondrial haplotypes that they bear (NML-1 or NML-II), there is only one taxon in the area, we describe it as: Iberolacerta monticola astur ssp. nov.Concerning the relationships of I. m. astur ssp. nov. with I. monticola s. str. and I. gal ani ("extra zonal analysis"), in the female analyses the new taxon centroid is closer to I. monticola s. str. than to I. gal ani (more similarity with I manticolas.str.), whereas in the male analyses the relationship is just the contrary (closer to I. gal ani, paralleling the direction of the hypothesized past hybridization). Moreover, in both sexes' ANOVA, I. m. astur ssp. nov. results more similar (lessP<0.05 differences) to I. galani than to I. monticola s. str. Osteologically, I. m. astur ssp. nov. is slightly more similar toI. monticola s. str. than to I. galani, especially in the squamosal bone, which is regularly arched (primitive shape). Genetically,as indicated above, the NML populations can be subdivided in two groups according to their mitochondrial DNA,namely NML-I (bearing clearly differentiated haplotypes, phylogenetically closer to I. monticola) and NML-II (whose haplotypes could have been mistaken for those of an I. gal ani population). This mitochondrial subdivision has at most a subtle nuclear correlate, however. According to the nuclear microsatellite markers, all the NML populations belong to a single group(/. m. astur ssp. nov.), which would be more similar to I. gal ani than to I monticola, with NML-II populations lying closer to I. galani than those from the NML-I group and, correspondingly, more distant from I. monticola. The discordant phylogenetic signal of mitochondrial and nuclear markers is discussed in terms of past introgression events and sex-biases in phylopatry and dispersion in these species. Iberolacerta manti cola astur ssp. nov., inhabits the Northern Montes de Leon (Sierra de Gistreo sensu latissimo ): Gistredo,Catoute, Tambaron, Nevadfn, Villabandfn (or Macizo del Alto de Ia Canada), Arcos del Agua (or Fernan Perez),Tiendas and Suspiron, mainly in quartzite and slate rock substrates. Its current distribution, cornered in the NW of theNorthern part of the Montes de Leon, suggests a possible competitive exclusion between this taxon and/. galani, as the galani haplotypes (NML-II) appear cornered in the most harsh and continental areas, speaking also about a, even in the past, very limited presence of this species in the area that probably was soon absorbed by I. m. astur ssp. nov. (with NMLI haplotypes). Variation in watershed limits (especially with l montico/a s. str. in the North) and Pleistocene climatic oscillations(with I. gal ani in the South) probably played a crucial role in isolation of the different Iberolacerta colonizationwaves in this zone. These changes in the boundaries among watersheds limited the contact between the NML and the main Cantabrian Mountains, restricting to narrow points (different along time) the contact between the two ranges, and thus,the areas for possible contact between I. m. astur ssp. nov. and I. monticola s. str. (see Fig. lB). The origin of this tax on dates back to the end of Pliocene or Lower Pleistocene (around 1.8 Mya), according to mtDNA divergence. On the other side, climatic oscillations allowed expansion and contact with the more continental harsh climate-dwelling I. gal ani.