Jiří Moravec

Phylogeography of the Middle Eastern tree frogs (Hyla, Hylidae, Amphibia) as inferred from nuclear and mitochondrial DNA variation, with a description of a new species

Evolutionary relationships of the tree frogs from the Middle East and the demographic histories of their populations were studied using a combination of mitochondrial and nuclear genes. Hyla savignyi and neighboring populations of H. orientalis (former eastern populations of H. arborea) were the main focus taxa. Within H. savignyi, a deep phylogenetic divergence dated about 8.4Ma was discovered. Southern populations from Yemen, Jordan, southern Syria and extreme north-eastern Israel are hereby described as a new species, H. felixarabica sp. nov. Our study points to a biogeographic connection of the south-western Arabian Peninsula and southern Levant and to the importance of the Dead Sea Rift as a historical barrier geographically separating the new species from H. savignyi. Major genetic breaks revealed within species (H. felixarabica: Yemen vs. Jordan-Syria; H. savignyi sensu stricto: Levant vs. Turkey-Iran) are probably connected to climate changes during the Plio-Pleistocene boundary, while the finer phylogeographic structuring probably resulted from the Quaternary climate oscillations. The Cypriote population of H. savignyi originated from southern Anatolia relatively recently. Hyla orientalis from the southern Black Sea region seems to be genetically quite uniform, although two phylogeographic units with western Turkish and Caucasus-Caspian affinities might be detected. Hyla savignyi and H. orientalis carry signals of population expansions dated to the middle to late Pleistocene, while populations of H. felixarabica seem to have rather been constant in size, which might indicate more stable climatic conditions in the southern regions during the Quaternary.

Slow worm, Anguis fragilis (Reptilia: Anguidae) as a species complex: Genetic structure reveals deep divergences

Phylogenetic relationships of the Western Palearctic legless lizard genus Anguis were inferred based on a fragment of mitochondrial DNA and two nuclear protein-coding loci, C-mos and PRLR. A. cephallonica from the Peloponnese was confirmed as a valid species. It is the sister taxon to a clade comprising all other evolutionary lineages, which were shown to represent three distinct species: (1) A. fragilis sensu stricto occurring in Western and Central Europe, the north-western Balkans, with possibly isolated populations in the eastern Balkans, and presumably also in western Scandinavia and Italy; (2) A. colchica distributed from the eastern Czech Republic and the Baltic region eastward to northern Iran, presumably also in eastern Scandinavia, and the north-eastern Balkans; (3) A. graeca restricted to the southern Balkans, and partially sympatric with A. cephallonica. According to the more variable mitochondrial marker, A. graeca appears to be the sister species to A. colchica, and these taxa together form a sister clade to A. fragilis, whereas the less variable nuclear markers show A. colchica to be closer to A. fragilis. The C-mos gene has not provided substantial variation within this species complex, while the PRLR gene, which was used for the first time in phylogeographic study in a reptile, distinguished all species successfully. Intra-specific differentiation of A. colchica is discussed, and subspecific status of the Caucasian and Caspian populations is proposed. The uncovered genetic differences should be taken into account in all future biogeographical, morphological and ecological studies, as well as in conservation.

Neogene climatic oscillations shape the biogeography and evolutionary history of the Eurasian blindsnake.

Typhlops vermicularis is the only extant scolecophidian representative occurring in Europe. Its main distribution area, the eastern Mediterranean, has a complicated geological and climatic history that has left an imprint on the phylogenies and biogeography of many taxa, especially amphibians and reptiles. Since reptiles are sensitive indicators of palaeogeographical and palaeoclimatic events, we investigated the intraspecific genealogy of T. vermicularis in a phylogeographical framework. A total of 130 specimens were analyzed, while the use of formalin and ethanol as preservatives called for a special treatment of the samples. Partial sequences of two mitochondrial (12S and ND2) and one nuclear (PRLR) marker were targeted and the results of the phylogenetic analyses (NJ, ML and BI) and the parsimony-network revealed the existence of 10 evolutionary significant units within this species. In combination with the results of the dispersal-vicariance analysis, we may conclude that the Eurasian blindsnake has encountered a sequence of extinction events, followed by secondary expansion from refugia. Estimation of divergence times showed that severe climatic changes between significantly wetter and drier conditions in the Late Neogene have played a key role on the evolutionary and biogeographical history of T. vermicularis. Additionally, both markers (mtDNA and nDNA) distinguished a largely-differentiated evolutionary lineage (Jordan and south Syria), which could even be reckoned as a full species. Our study reveals the existence of cryptic evolutionary lineages within T. vermicularis, which calls for further attention both on the protection of intraspecific varieties and the respective geographic areas that hold them.

An ancient lineage of slow worms, genus Anguis (Squamata: Anguidae), survived in the Italian Peninsula

Four species of legless anguid lizard genus Anguis have been currently recognized: A. fragilis from western and central Europe, A. colchica from eastern Europe and western Asia, A. graeca from southern Balkans, and A. cephallonica from the Peloponnese. Slow worms from the Italian Peninsula have been considered conspecific with A. fragilis, despite the fact that the region served as an important speciation center for European flora and fauna, and included some Pleistocene glacial refugia. We used mitochondrial and nuclear DNA sequences to investigate the systematic and phylogenetic position of the Italian slow-worm populations and morphological analyses to test for phenotypic differentiation from A. fragilis from other parts of Europe. Our phylogenetic analyses revealed that Italian slow worms form a distinct deeply differentiated mtDNA clade, which presumably diverged during or shortly after the basal radiation within the genus Anguis. In addition, the specimens assigned to this clade bear distinct haplotypes in nuclear PRLR gene and show morphological differentiation from A. fragilis. Based on the differentiation in all three independent markers, we propose to assign the Italian clade species level under the name Anguis veronensisPollini, 1818. The newly recognized species is distributed throughout the Italian Peninsula to the Southern Alps and south-eastern France. We hypothesize that the Tertiary Alpine orogeny with subsequent vicariance might have played a role in differentiation of this species. The current genetic variability was later presumably shaped in multiple glacial refugia within the Italian Peninsula, with the first splitting event separating populations from the region of the Dolomite Mountains.

Out of Africa: Phylogeny and biogeography of the widespread genus Acanthodactylus (Reptilia: Lacertidae)

Acanthodactylus lizards are among the most diverse and widespread diurnal reptiles in the arid regions spanning from North Africa across to western India. Acanthodactylus constitutes the most species-rich genus in the family Lacertidae, with over 40 recognized species inhabiting a wide variety of dry habitats. The genus has seldom undergone taxonomic revisions, and although there are a number of described species and species-groups, their boundaries, as well as their interspecific relationships, remain largely unresolved. We constructed a multilocus phylogeny, combining data from two mitochondrial (12S, cytb) and three nuclear (MC1R, ACM4, c-mos) markers for 302 individuals belonging to 36 known species, providing the first large-scale time-calibrated molecular phylogeny of the genus. We evaluated phylogenetic relationships between and within species-groups, and assessed Acanthodactylus biogeography across its known range. Acanthodactylus cladogenesis is estimated to have originated in Africa due to vicariance and dispersal events from the Oligocene onwards. Radiation started with the separation into three clades: the Western and scutellatus clades largely distributed in North Africa, and the Eastern clade occurring mostly in south-west Asia. Most Acanthodactylus species diverged during the Miocene, possibly as a result of regional geological instability and climatic changes. We support most of the current taxonomic classifications and phylogenetic relationships, and provide genetic validity for most species. We reveal a new distinct blanfordii species-group, suggest new phylogenetic positions (A. hardyi, A. masirae), and synonymize several species and subspecies (A. lineomaculatus, A. boskianus khattensis and A. b. nigeriensis) with their phylogenetically closely-related species. We recommend a thorough systematic revision of taxa, such as A. guineensis, A. grandis, A. dumerilii, A. senegalensis and the pardalis and erythrurus species-groups, which exhibit high levels of intraspecific variability, and clear evidence of phylogenetic complexity.

Contrasting evolutionary histories of the legless lizards slow worms (Anguis) shaped by the topography of the Balkan Peninsula

BackgroundGenetic architecture of a species is a result of historical changes in population size and extent of distribution related to climatic and environmental factors and contemporary processes of dispersal and gene flow. Population-size and range contractions, expansions and shifts have a substantial effect on genetic diversity and intraspecific divergence, which is further shaped by gene-flow limiting barriers. The Balkans, as one of the most important sources of European biodiversity, is a region where many temperate species persisted during the Pleistocene glaciations and where high topographic heterogeneity offers suitable conditions for local adaptations of populations. In this study, we investigated the phylogeographical patterns and demographic histories of four species of semifossorial slow-worm lizards (genus Anguis) present in the Balkan Peninsula, and tested the relationship between genetic diversity and topographic heterogeneity of the inhabited ranges.ResultsWe inferred phylogenetic relationships, compared genetic structure and historical demography of slow worms using nucleotide sequence variation of mitochondrial DNA. Four Anguis species with mostly parapatric distributions occur in the Balkan Peninsula. They show different levels of genetic diversity. A signature of population growth was detected in all four species but with various courses in particular populations. We found a strong correlation between genetic diversity of slow-worm populations and topographic ruggedness of the ranges (mountain systems) they inhabit. Areas with more rugged terrain harbour higher genetic diversity.ConclusionsPhylogeographical pattern of the genus Anguis in the Balkans is concordant with the refugia-within-refugia model previously proposed for both several other taxa in the region and other main European Peninsulas. While slow-worm populations from the southern refugia mostly have restricted distributions and have not dispersed much from their refugial areas, populations from the extra-Mediterranean refugia in northern parts of the Balkans have colonized vast areas of eastern, central, and western Europe. Besides climatic historical events, the heterogeneous topography of the Balkans has also played an important role in shaping genetic diversity of slow worms.

Hidden relationships and genetic diversity: Molecular phylogeny and phylogeography of the Levantine lizards of the genus Phoenicolacerta (Squamata: Lacertidae)

The Levant region witnessed dramatic tectonic events and climatic fluctuations that changed the historical landscape of the area and consequently influenced the cladogenesis and distribution of the local biota. In this study we use information from two mitochondrial and two nuclear genes and species delimitation methods in order to obtain the first robust time-calibrated molecular phylogeny of the Levantine rock lizards of the genus Phoenicolacerta. We sampled from across its distributional range with the aim to clarify its systematics, biogeography and evolution. Our results suggest that the genus includes two well-supported clades, one comprising solely the montane species Phoenicolacerta kulzeri, and the other including the three remaining species, the relatively widespread, P. laevis, the Syrian-Turkish P. cyanisparsa and the Cypriot endemic P. troodica. We found that both P. laevis and P. cyanisparsa are not monophyletic, as the Turkish populations of P. laevis branch within P. cyanisparsa. We found high levels of undescribed diversity within P. laevis which necessitate a thorough revision. We suggest that Phoenicolacerta started radiating during the mid-late Miocene, and that both vicariance and dispersal events shaped the diversification and distribution of the genus concomitantly with the formation of major geological structures and climatic fluctuations in the Levant. These results highlight the region as an important center of speciation, contributing to the species diversity of the eastern Mediterranean.

Pleistocene extinctions and recent expansions in an anguid lizard of the genus Pseudopus

Climatic changes during the Pleistocene played an instrumental role in the shaping recent distribution and diversity of the Western Palearctic biota. Range oscillations often lead to allopatric differentiation followed by the establishment of secondary contact zones. As a result, many species are composed of complex networks of phylogenetic lineages with different histories. Pseudopus apodus is the only surviving member of an ancient genus of Western Palearctic anguid lizards (Anguidae) distributed from the Balkans through Anatolia and Caucasus to central Asia. Here, we used mitochondrial and nuclear DNA sequences to reconstruct the evolutionary history of the species in phylogeographic and demographic frameworks. Our analyses revealed three main phylogenetic lineages that diverged during or shortly before the Pleistocene. Two of them more or less correspond to the known subspecies, and their low genetic variability suggests relatively recent dispersal and colonization of vast parts of the range. The third, southern, lineage is more geographically restricted and diversified than the other two. This pattern shows that the Quaternary climatic oscillations presumably caused repeated large‐scale population extinctions of the species, depleting most of its diversity. Only a few refugia located in Anatolia, Levant, and Transcaucasia served as sources for subsequent recolonization to the areas of the recent distribution. This is in contrast to many other Western Palearctic reptiles that survived unfavorable climatic conditions in numerous local refugia and sanctuaries, which resulted in more complex phylogenetic structure.

Cutting the Gordian Knot: Phylogenetic and ecological diversification of the Mesalina brevirostris species complex (Squamata, Lacertidae)

Mesalina are small lacertid lizards occurring in the Saharo‐Sindian deserts from North Africa to the east of the Iranian plateau. Earlier phylogenetic studies indicated that there are several species complexes within the genus and that thorough taxonomic revisions are needed. In this study, we aim at resolving the phylogeny and taxonomy of the M. brevirostris species complex distributed from the Middle East to the Arabian/Persian Gulf region and Pakistan. We sequenced three mitochondrial and three nuclear gene fragments, and in combination with species delimitation and species‐tree estimation, we infer a time‐calibrated phylogeny of the complex. The results of the genetic analyses support the presence of four clearly delimited species in the complex that diverged approximately between the middle Pliocene and the Pliocene/Pleistocene boundary. Species distribution models of the four species show that the areas of suitable habitat are geographically well delineated and nearly allopatric, and that most of the species have rather divergent environmental niches. Morphological characters also confirm the differences between the species, although sometimes minute. As a result of all these lines of evidence, we revise the taxonomy of the Mesalina brevirostris species complex. We designate a lectotype for Mesalina brevirostris Blanford, 1874; resurrect the available name Eremias bernoullii Schenkel, 1901 from the synonymy of M. brevirostris; elevate M. brevirostris microlepis (Angel, 1936) to species status; and describe Mesalina saudiarabica, a new species from Saudi Arabia.