Major mtDNA Lineages Research Articles

River Drainage Reorganization and Reticulate Evolution in the Two-Lined Salamander (Eurycea bislineata) Species Complex.

The origin and eventual loss of biogeographic barriers can create alternating periods of allopatry and secondary contact, facilitating gene flow among distinct metapopulations and generating reticulate evolutionary histories that are not adequately described by a bifurcating evolutionary tree. One such example may exist in the two-lined salamander (Eurycea bislineata) species complex, where discordance among morphological and molecular datasets has created a "vexing taxonomic challenge." Previous phylogeographic analyses of mitochondrial DNA (mtDNA) suggested that the reorganization of Miocene paleodrainages drove vicariance and dispersal, but the inherent limitations of a single-locus dataset precluded the evaluation of subsequent gene flow. Here, we generate triple-enzyme restriction site-associated DNA sequencing (3RAD) data for > 100 individuals representing all major mtDNA lineages and use a suite of complementary methods to demonstrate that discordance among earlier datasets is best explained by a reticulate evolutionary history influenced by river drainage reorganization. Systematics of such groups should acknowledge these complex histories and relationships that are not strictly hierarchical. [Amphibian; hybridization; introgression; Plethodontidae; stream capture.].

Phylogeography and Genetic Structure in the California Giant Salamander (Dicamptodon ensatus): Impacts of current and historic landscape features.

The California Floristic Province contains numerous ecological regions and a complex geological and geographical history that make it one of the worlds biodiversity hotspots. A number of wide-ranging taxa span across these regions and show complex patterns of dispersal, vicariance and lineage diversification, making localized small ranged species with lower levels of vagility essential to understanding the overall region. Here, we investigate the biogeography and population structure of the California Giant Salamander (Dicamptodon ensatus) (Eschscholtz 1833), an endemic species localized to a narrow coastal region between two areas of biological significance in the California Floristic Province, the North Coast Divide and Monterey Bay. We sequenced one mtDNA fragment (control region) for 133 individuals and a subset of 38 individuals for the anonymous nuclear locus E16C7. We analyzed these sequences with phylogenetic, coalescent, Bayesian clustering, and population genetic approaches in order to infer population structure, phylogenetic structure, and biogeographic history. Additionally, we examined occurrence data with species distribution modeling to generate a habitat suitability map to aid our interpretation of geographic structure. Our analyses recovered 4 major mtDNA lineages, two of which are combined into 3 major lineages when nuDNA is examined. These 3 major lineages are bounded by 4 major current or past geological features; the North Coast Divide, the former Wilson Grove Embayment/current Petaluma Gap, San Francisco Bay, and Monterey Bay. Other low-vagility species linked to moist microclimates and forest habitat do share similarities with the genetic patterns of D. ensatus hinting at a larger role for the past Wilson Grove embayment and modern Petaluma Gap in California biogeography.

Phylogeography of the Brittle Star Ophiura sarsii Lütken, 1855 (Echinodermata: Ophiuroidea) from the Barents Sea and East Atlantic

Ophiura sarsii is a common brittle star species across the Arctic and Sub-Arctic regions of the Atlantic and the Pacific oceans. Ophiurasarsii is among the dominant echinoderms in the Barents Sea. We studied the genetic diversity of O.sarsii by sequencing the 548 bp fragment of the mitochondrial COI gene. Ophiurasarsii demonstrated high genetic diversity in the Barents Sea. Both major Atlantic mtDNA lineages were present in the Barents Sea and were evenly distributed between the northern waters around Svalbard archipelago and the southern part near Murmansk coast of Kola Peninsula. Both regions, and other parts of the O.sarsii range, were characterized by high haplotype diversity with a significant number of private haplotypes being mostly satellites to the two dominant haplotypes, each belonging to a different mtDNA clade. Demographic analyses indicated that the demographic and spatial expansion of O.sarsii in the Barents Sea most plausibly has started in the Bølling–Allerød interstadial during the deglaciation of the western margin of the Barents Sea.

Regional‐scale aquifer hydrogeology as a driver of phylogeographic structure in the Neotropical catfish Rhamdia guatemalensis (Siluriformes: Heptapteridae) from cenotes of the Yucatán Peninsula, Mexico

Abstract The Yucatán Peninsula (YP) contains one of the most spectacular and developed karstic aquifers in the world, which connects to the surface through water‐filled sinkholes locally known as cenotes. Cenotes and their associated submerged caves are home to a remarkable diversity of aquatic fauna, yet very little is known about spatial patterns of intraspecific genetic and morphological variation across the aquifer. The strong affinity to cenotes (vs. submerged caves) exhibited by most fishes inhabiting the YP aquifer raises a fundamental question about the ecology and evolutionary history of its ichthyofauna: how genetically structured and morphologically divergent are fish populations from different cenotes across the YP? To shed light on this question, we investigated phylogeographic structure in the Neotropical catfish Rhamdia guatemalensis, arguably the most ubiquitous representative of the YP aquifer ichthyofauna. Geographic patterns of genetic variation were determined through phylogeographic and population genetics analyses of mitochondrial (mt)DNA sequence data. Body shape variation was investigated using both traditional and geometric morphometrics. Dense sampling across the previously overlooked YP region, coupled with the inclusion of samples from basins unrepresented in past studies, allowed for revision of previous estimates of phylogeographic structuring across the species’ range. Our results reveal a striking spatial correspondence between major mtDNA lineages and physiographic regions defined on the basis of broad‐scale patterns of groundwater flow. Morphological variation in populations from the YP, although considerable, neither reflects the observed spatial signal of genetic differentiation nor displays any other discernible geographic pattern. The resulting range‐wide phylogeographic pattern is largely consistent with a latitudinal, isolation‐by‐distance gradient, and the expectations from catchment hydrology, while revealing an unexpectedly close relationship between YP aquifer and epigean populations (both inside and outside the YP). Although the inferred range‐wide phylogeographic history is consistent with a single aquifer colonisation event followed by multiple instances of dispersal out of the aquifer and into neighbouring epigean basins, hypotheses regarding the colonisation and exodus of the YP by R. guatemalensis require further investigation. Within the YP aquifer, regional‐scale hydrogeological differences might impose limits to underground dispersal, potentially driving genetic divergence in populations of R. guatemalensis, and possibly in other cenote‐dwelling fishes as well. Future research focused on other YP aquifer species, however, is needed to test the generality of our findings. Phylogeographic research on the stygofauna of the YP aquifer is a relatively nascent field of inquiry. Besides shedding novel light into the patterns and drivers of genetic differentiation in fish populations from across the aquifer, this study sets the stage for future research aimed at unravelling the evolutionary history of the stygofauna that inhabits the cenotes and submerged caves of the YP. The remarkable and hitherto undocumented phylogeographic pattern revealed herein (i.e. correspondence of major mtDNA lineages with physiographic regions defined by regional‐scale fracture zones) adds to a growing body of evidence in support of the notion that the architecture of freshwater aquifers may include physical barriers that can act as regional‐scale drivers of groundwater faunal distributions and genetic structuring.

Distribution of genetic diversity reveals colonization patterns and philopatry of the loggerhead sea turtles across geographic scales

Understanding the processes that underlie the current distribution of genetic diversity in endangered species is a goal of modern conservation biology. Specifically, the role of colonization and dispersal events throughout a species’ evolutionary history often remains elusive. The loggerhead sea turtle (Caretta caretta) faces multiple conservation challenges due to its migratory nature and philopatric behaviour. Here, using 4207 mtDNA sequences, we analysed the colonisation patterns and distribution of genetic diversity within a major ocean basin (the Atlantic), a regional rookery (Cabo Verde Archipelago) and a local island (Island of Boa Vista, Cabo Verde). Data analysis using hypothesis-driven population genetic models suggests the colonization of the Atlantic has occurred in two distinct waves, each corresponding to a major mtDNA lineage. We propose the oldest lineage entered the basin via the isthmus of Panama and sequentially established aggregations in Brazil, Cabo Verde and in the area of USA and Mexico. The second lineage entered the Atlantic via the Cape of Good Hope, establishing colonies in the Mediterranean Sea, and from then on, re-colonized the already existing rookeries of the Atlantic. At the Cabo Verde level, we reveal an asymmetric gene flow maintaining links across island-specific nesting groups, despite significant genetic structure. This structure stems from female philopatric behaviours, which could further be detected by weak but significant differentiation amongst beaches separated by only a few kilometres on the island of Boa Vista. Exploring biogeographic processes at diverse geographic scales improves our understanding of the complex evolutionary history of highly migratory philopatric species. Unveiling the past facilitates the design of conservation programmes targeting the right management scale to maintain a species’ evolutionary potential.

Integrative taxonomic investigation ofPetaurus breviceps(Marsupialia: Petauridae) reveals three distinct species

AbstractThe Australian sugar glider, Petaurus breviceps s.l., is widely distributed across eastern and northern Australia. Examination of historical and contemporary collections of Petaurus specimens and phylogenetic analyses have revealed considerable taxonomic diversity within the genus. We aimed to utilize an integrative taxonomic approach, combining genetic and morphological evidence, to resolve the taxonomy of Australian gliders currently recognized as Petaurus breviceps. Herein, we confirm the existence of three distinct species: P. breviceps, P. notatus comb. nov. and P. ariel comb. nov.. Petaurus breviceps and P. notatus are each represented by major mtDNA lineages in P. breviceps, while P. ariel forms a sister-lineage to P. norfolcensis and P. gracilis. Subtle morphological differences distinguish P. breviceps from the closely related P. notatus, while the morphological distinctions between P. ariel and its genetically similar sister-taxa, P. norfolcensis and P. gracilis, are more obvious. Given the purported broad geographic distribution of the taxon, P. breviceps s.l. was not listed as threatened, but dividing this taxon into three species has important conservation implications for all taxa in the group, particularly given the lamentable record for mammal extinctions in Australia. Concerted and targeted conservation efforts are necessary to preserve these distinct, newly described species.

Global phylogeography reveals the origin and the evolutionary history of the gypsy moth (Lepidoptera, Erebidae)

We examined the global phylogeography of gypsy moth (Lymantria dispar L.) using molecular data based on mitochondrial and nuclear genes. Populations from all biogeographic regions of the native and introduced range of L. dispar, were sampled to fully document intraspecific and subspecies variation, identify potential cryptic species, and to clarify the relationships among major phylogeographic lineages. We recovered three major mtDNA lineages of L. dispar: Transcaucasia; East Asia + Japan; and Europe + Central Asia. The circumscription of these lineages is only partially consistent with the current taxonomic concept (i.e., L. dispar dispar; L. dispar asiatica; L. dispar japonica), with the following important discrepancies: (1) north-central Asian populations, including topotypical populations of L. dispar asiatica, may be more closely related to European rather than Asian segregates, which would require the synonymization of the taxon asiatica and establishment of a new name; (2) the Japanese populations (L. d. japonica) are not distinct from east Asian populations; (3) the presence of a distinct, unnamed mitogenomic lineage endemic to the Trancaucasus region. We demonstrated that the population from Transcaucasia contains the highest mitochondrial haplotype diversity among L. dispar, potentially indicative of an ancestral area for the entire dispar-group. Our study corroborates the endemic Hokkaido, Japan taxon Lymantria umbrosa (Butler) as the sister group to all other L. dispar populations, but the applicability of the names umbrosa versus hokkaidoensis Goldschmidt needs to be re-evaluated. The ancestral area analysis suggest that Japan was likely colonized via Sakhalin ∼1 Mya, in contrast to previous studies which have suggested colonization of the Japanese archipelago via the Korean Peninsula. Lastly, mitogenomic variation within L. dispar is incongruent with phylogenies based on nuclear DNA, as nDNA gene phylogenies did not recover the three major mtDNA lineages, and also failed to recover L. dispar and L. umbrosa as reciprocally monophyletic.

Plio-Pleistocene diversification and biogeographic barriers in southern Australia reflected in the phylogeography of a widespread and common lizard species

Palaeoclimatic events and biogeographical processes since the mid-Tertiary have played an important role in shaping the evolution and distribution of Australian fauna. However, their impacts on fauna in southern and arid zone regions of Australia are not well understood. Here we investigate the phylogeography of an Australian scincid lizard, Tiliqua rugosa, across southern Australia using mitochondrial DNA (mtDNA) and 11 nuclear DNA markers (nuDNA), including nine anonymous nuclear loci. Phylogenetic analyses revealed three major mtDNA lineages within T. rugosa, geographically localised north and south of the Murray River in southern Australia, and west of the Nullarbor Plain. Molecular variance and population analyses of both mtDNA and nuDNA haplotypes revealed significant variation among the three populations, although potential introgression of nuDNA markers was also detected for the Northern and Southern population. Coalescent times for major mtDNA lineages coincide with an aridification phase, which commenced after the early Pliocene and increased in intensity during the Late Pliocene-Pleistocene. Species distribution modelling and a phylogeographic diffusion model suggest that the range of T. rugosa may have contracted during the Last Glacial Maximum and the locations of optimal habitat appear to coincide with the geographic origin of several distinct mtDNA lineages. Overall, our analyses suggest that Plio-Pleistocene climatic changes and biogeographic barriers associated with the Nullarbor Plain and Murray River have played a key role in shaping the present-day distribution of genetic diversity in T. rugosa and many additional ground-dwelling animals distributed across southern Australia.

Differentiation underground: Range-wide multilocus genetic structure of the silvery mole-rat does not support current taxonomy based on mitochondrial sequences

The silvery mole-rat (Heliophobius argenteocinereus) is a solitary subterranean rodent with its distribution centred mainly in miombo woodlands of eastern Africa. This part of the continent was significantly influenced by the formation of the East African Rift System (EARS) during the last 25 Mya and by pronounced climatic changes in the Plio-Pleistocene that have caused genetic differentiation leading even to speciation events in many organisms. Recently, based on analysis of mitochondrial (mt) DNA sequences, it was suggested that H. argenteocinereus is a complex of six to eight species that diverged from the early to middle Miocene. In the present study, we significantly extended the sampling, re-analysed mtDNA datasets and analysed nuclear markers with the aim to assess the evolutionary history of Heliophobius. If we do not consider the old museum samples from south-eastern Democratic Republic of Congo (very divergent short mtDNA sequences obtained from ancient DNA, requiring further study from fresh material), the genus Heliophobius is composed of three major mtDNA lineages with parapatric distribution. The Rukwa Rift (+Mbeya triple junction) at the Zambia-Tanzania border, Lake Malawi, and the Eastern Arc Mountains form the biogeographical divides among these clades. The relatively shallow differences among the mitochondrial clades, divergence dating based on the use of the fossilised birth-death ratio model and a multi-locus dataset, and a very similar pattern of genetic structure to other rodents inhabiting the same area and habitat, suggest that the evolutionary history of the extant silvery mole-rat was predominantly influenced by the climatic fluctuations in the Plio-Pleistocene. Awaiting further studies employing genomic, ecological, morphological or behavioural data, we advocate for using the single name H. argenteocinereus for all evolutionary lineages within this taxon, because (1) comparison of the genetic structure observed in mtDNA and nuclear markers suggest hybridization between at least some mtDNA lineages in the contact zones; and (2) new samples close to the type localities suggest incorrect use of previous names.

Cryptic diversity in Amazonian frogs: Integrative taxonomy of the genus Anomaloglossus (Amphibia: Anura: Aromobatidae) reveals a unique case of diversification within the Guiana Shield

Lack of resolution on species boundaries and distribution can hamper inferences in many fields of biology, notably biogeography and conservation biology. This is particularly true in megadiverse and under-surveyed regions such as Amazonia, where species richness remains vastly underestimated. Integrative approaches using a combination of phenotypic and molecular evidence have proved extremely successful in reducing knowledge gaps in species boundaries, especially in animal groups displaying high levels of cryptic diversity like amphibians. Here we combine molecular data (mitochondrial 16S rRNA and nuclear TYR, POMC, and RAG1) from 522 specimens of Anomaloglossus, a frog genus endemic to the Guiana Shield, including 16 of the 26 nominal species, with morphometrics, bioacoustics, tadpole development mode, and habitat use to evaluate species delineation in two lowlands species groups. Molecular data reveal the existence of 18 major mtDNA lineages among which only six correspond to described species. Combined with other lines of evidence, we confirm the existence of at least 12 Anomaloglossus species in the Guiana Shield lowlands. Anomaloglossus appears to be the only amphibian genus to have largely diversified within the eastern part of the Guiana Shield. Our results also reveal strikingly different phenotypic evolution among lineages. Within the A. degranvillei group, one subclade displays acoustic and morphological conservatism, while the second subclade displays less molecular divergence but clear phenotypic divergence. In the A. stepheni species group, a complex evolutionary diversification in tadpole development is observed, notably with two closely related lineages each displaying exotrophic and endotrophic tadpoles.

Phylogeography of the arid-adapted Malagasy bullfrog, Laliostoma labrosum, influenced by past connectivity and habitat stability

The rainforest biome of eastern Madagascar is renowned for its extraordinary biodiversity and restricted distribution ranges of many species, whereas the arid western region of the island is relatively species poor. We provide insight into the biogeography of western Madagascar by analyzing a multilocus phylogeographic dataset assembled for an amphibian, the widespread Malagasy bullfrog, Laliostoma labrosum. We find no cryptic species in L. labrosum (maximum 1.1% pairwise genetic distance between individuals in the 16S rRNA gene) attributable to considerable gene flow at the regional level as shown by genetic admixture in both mtDNA and three nuclear loci, especially in central Madagascar. Low breeding site fidelity, viewed as an adaptation to the unreliability of standing pools of freshwater in dry and seasonal environments, and a ubiquitous distribution within its range may underlie overall low genetic differentiation. Moreover, reductions in population size associated with periods of high aridity in western Madagascar may have purged DNA variation in this species. The mtDNA gene tree revealed seven major phylogroups within this species, five of which show mostly non-overlapping distributions. The nested positions of the northern and central mtDNA phylogroups imply a southwestern origin for all extant mtDNA lineages in L. labrosum. The current phylogeography of this species and paleo-distributions of major mtDNA lineages suggest five potential refugia in northern, western and southwestern Madagascar, likely the result of Pleistocene range fragmentation during drier and cooler climates. Lineage sorting in mtDNA and nuclear loci highlighted a main phylogeographic break between populations north and south of the Sambirano region, suggesting a role of the coastal Sambirano rainforest as a barrier to gene flow. Paleo-species distribution models and dispersal networks suggest that the persistence of some refugial populations was mainly determined by high population connectivity through space and time.

Phylogeography of the narrow-headed vole Lasiopodomys (Stenocranius ) gregalis (Cricetidae, Rodentia) inferred from mitochondrial cytochrome b sequences: an echo of Pleistocene prosperity

A species-wide phylogeographic study of the narrow-headed vole Lasiopodomys (Stenocranius) gregalis was performed using the mitochondrial (mt) cytochrome b gene. We examined 164 specimens from 50 localities throughout the species distribution range. Phylogeographic pattern clearly demonstrates the division into four major mtDNA lineages with further subdivision. The level of genetic differentiation between them was found to be extremely high even for the species level: about 6–11%. The most striking result of our study is extremely high mutation rate of cytb in L. gregalis. Our estimates suggested its value of 3.1 × 10−5 that is an order of magnitude higher than previous estimates for Microtus species. The mean estimated time of basal differentiation of the narrow-headed vole is about 0.8 Mya. This time estimate is congruent with the known paleontological record. The greatest mitochondrial diversity is found in Southern Siberia where all four lineages occur; therewith, three of them are distributed exclusively in that area. The lineage that is distributed in south-eastern Transbaikalia is the earliest derivate and exhibits the highest genetic divergence from all the others (11%). It is quite probable that with further research, this lineage will turn out to represent a cryptic species. Spatial patterns of genetic variation in populations of the narrow-headed vole within the largest mt lineage indicate the normal or stepping stone model of dispersal to the north and south-west from the Altay region in Middle Pleistocene. Both paleontological data and genetic diversity estimates suggest that this species was very successful during most of the Pleistocene, and we propose that climate humidification and wide advance of tree vegetation at the Pleistocene–Holocene boundary promoted range decrease and fragmentation for this typical member of tundra-steppe faunistic complex. However, we still observe high genetic diversity within isolated fragments of the range.

Fine-scale phylogeographic contact zone in Austrian brown trout Salmo trutta reveals multiple waves of post-glacial colonization and a pre-dominance of natural versus anthropogenic admixture

Two major mtDNA lineages of brown trout come into contact in the upper Danube basin of Central Europe. The region is additionally confronted with stock management programs that often use domesticated foreign strains creating confusion as to what constitutes native and non-native populations as well as how to define major management units. In a unique system in north eastern Austria we provide additional support that the so-called Atlantic lineage of brown trout is native to the upper Danube and provide a high resolution microsatellite screening protocol that clearly distinguishes between natural and anthropogenic introgression, with the latter being minimal. Sequencing of 6,000 bp of mtDNA also shows that domesticated stocks primarily stem from a distinct albeit genetically diverse sub-clade of the Atlantic lineage, approximately 50,000–100,000 years divergent from the native Atlantic stocks, thought to have expanded into the region during the Holocene. The futility in defining large-scale management units based on mtDNA lineages for such broadly distributed and highly fragmented species is discussed.

Holarctic genetic structure and range dynamics in the woolly mammoth

Ancient DNA analyses have provided enhanced resolution of population histories in many Pleistocene taxa. However, most studies are spatially restricted, making inference of species-level biogeographic histories difficult. Here, we analyse mitochondrial DNA (mtDNA) variation in the woolly mammoth from across its Holarctic range to reconstruct its history over the last 200 thousand years (kyr). We identify a previously undocumented major mtDNA lineage in Europe, which was replaced by another major mtDNA lineage 32–34 kyr before present (BP). Coalescent simulations provide support for demographic expansions at approximately 121 kyr BP, suggesting that the previous interglacial was an important driver for demography and intraspecific genetic divergence. Furthermore, our results suggest an expansion into Eurasia from America around 66 kyr BP, coinciding with the first exposure of the Bering Land Bridge during the Late Pleistocene. Bayesian inference indicates Late Pleistocene demographic stability until 20–15 kyr BP, when a severe population size decline occurred.

Genetic population structure in Siniperca scherzeri (Perciformes: Siniperca) in China inferred from mitochondrial DNA sequences and microsatellite loci

This study aims to improve the estimates of fine-scale genetic diversity and to compare the population structure of Siniperca scherzeri with nuclear and mitochondrial DNA (mtDNA) markers. We first combined mtDNA sequences and seven microsatellite loci to examine the genetic diversity of S. scherzeri in China. Phylogenetic and nested clade analyses revealed two major mtDNA lineages and four subclades. Bayesian multilocus genotype clustering from the microsatellite loci revealed that regional divergence estimates were quantitatively congruent between marker classes. These lineages should be recognized as the basic evolutionary significant units for S. scherzeri in China. Contradict to the previous studies on Coreoperca whiteheadi and Siniperca chuatsi, neither the Nanling–Wuyi Mountain range nor the Qingling–Dabie Mountain range represented a major phylogeographic barrier for S. scherzeri. Our fine-scale analyses of the genetic population structure of S. scherzeri provided insights into the evolutionary processes that shaped the genetic heterogeneity of this fish. This study may serve as a basis for the protection, monitoring and breeding improvement of germplasm resources of this commercially important fish.

Genetic variation in brown trout Salmo truttaacross the Danube, Rhine, and Elbe headwaters: a failure of the phylogeographic paradigm?

BackgroundBrown trout Salmo trutta have been described in terms of five major mtDNA lineages, four of which correspond to major ocean basins, and one, according to some authors, to a distinct taxon, marbled trout Salmo marmoratus. The Atlantic and Danubian lineages of brown trout meet in a poorly documented contact zone in Central Europe. The natural versus human mediated origin of the Atlantic lineage in the upper Danube is a question of both theoretical and practical importance with respect to conservation management. We provide a comprehensive population genetic analysis of brown trout in the region with the aim of evaluating the geographic distribution and genetic integrity of these two lineages in and around their contact zone.ResultsGenetic screening of 114 populations of brown trout across the Danube/Rhine/Elbe catchments revealed a counter-intuitive phylogeographic structure with near fixation of the Atlantic lineage in the sampled portions of the Bavarian Danube. Along the Austrian Danube, phylogeographic informative markers revealed increasing percentages of Danube-specific alleles with downstream distance. Pure Danube lineage populations were restricted to peri-alpine isolates within previously glaciated regions. Both empirical data and simulated hybrid comparisons support that trout in non-glaciated regions north and northeast of the Alps have an admixed origin largely based on natural colonization. In contrast, the presence of Atlantic basin alleles south and southeast of the Alps stems from hatchery introductions and subsequent introgression. Despite extensive stocking of the Atlantic lineage, little evidence of first generation stocked fish or F1 hybrids were found implying that admixture has been established over time.ConclusionsA purely phylogeographic paradigm fails to describe the distribution of genetic lineages of Salmo in Central Europe. The distribution pattern of the Atlantic and Danube lineages is extremely difficult to explain without invoking very strong biological mechanisms.The peri-alpine distribution of relict populations of pure Danubian lineage brown trout implies that they colonized headwater river courses post-glacially ahead of the expansion of the Atlantic lineage. The recognition of natural as opposed to anthropogenic introgression of the Atlantic lineage into Danubian gene pools is of fundamental importance to management strategies.

Complete mitochondrial genomes resolve phylogenetic relationships within Bombina (Anura: Bombinatoridae)

A highly resolved and time-calibrated phylogeny based on nucleotide variation in 18 complete mitochondrial genomes is presented for all extant species and major lineages of fire-bellied toads of the genus Bombina (Bombinatoridae). Two sets of divergence time estimates are inferred by applying alternative fossil constraints as minima. Divergence time estimates from both analyses differed for the two oldest nodes. The earliest phylogenetic split occurred between small- and large-bodied Bombina (subgenera Bombina and Grobina, respectively) either in the Middle Oligocene or the Early Miocene. East Asian B. orientalis and European B. bombina+B. variegata diverged in the early or Middle Miocene. Divergence times inferred using the alternative fossil calibration strategies converged for the younger nodes, with broadly overlapping HPD intervals. The split between Bombina bombina and B. variegata occurred in the Late Miocene of Europe and somewhat preceded another deep mtDNA division between the Balkan B. v. scabra and B. v. variegata inhabiting the Carpathian Mts. Concurrently, the genetically distinct B. maxima diverged from other Grobina in southeast Asia in the Late Miocene or Pliocene. Our mtDNA phylogeny and a new species-tree analysis of published data (nuclear and mtDNA) suggest that B. fortinuptialis, B. lichuanensis and B. microdeladigitora may be conspecific geographic forms that separated due to Pleistocene climatic fluctuations in southeastern Asia. In the western Palearctic, the Late Pliocene to Pleistocene climatic vagaries most probably induced vicariant events in the evolutionary history of B. variegata that led to the formation of the two Balkan B. v. scabra lineages and the allopatric B. v. pachypus in the Apennine Peninsula. Divergence among B. bombina mtDNA lineages is low, with an Anatolian Turkey lineage as the sister group to the European mtDNA clades. In sum, Miocene diversification in the genus Bombina established six allopatrically distributed major mtDNA lineages that diversified during the Pliocene and Pleistocene and have survived until the present. The narrow habitat requirements of fire-bellied toads and extensive environmental changes throughout the Palearctic in the Neogene may have contributed to a putatively high extinction rate in these anurans resulting in the current east/west disjunction of their ranges.

From southern refugia to the northern range margin: genetic population structure of the common wall lizard, Podarcis muralis

AbstractAimThermophilic species persisted in southern refugia during the cold phases of the Pleistocene, and expanded northwards during warming. These processes caused genetic imprints, such as a differentiation of genetic lineages and a loss of genetic diversity in the wake of (re)colonization. We used molecular markers and species distribution models (SDMs) to study the impact of range dynamics on the common wall lizard, Podarcis muralis, from southern refugia to the northern range margin.LocationParts of the Western Palaearctic.MethodsWe genotyped 10 polymorphic microsatellites in 282 individuals of P. muralis and sequenced the mitochondrial DNA (mtDNA) cytochrome b gene to study the genetic structure, divergence times and ancestral distributions. Furthermore, we generated SDMs for climate scenarios for 6 and 21 ka derived from two different global circulation models.ResultsWe detected two major mtDNA lineages – a western France clade (Pyrenees to Brittany), and an eastern France clade (southern France to Germany, Belgium and Luxembourg). This split was dated to c. 1.23 Ma. The latter clade was divided into two subclades, which diverged c. 0.38 Ma. Genetic diversity of microsatellites within each clade was nested and showed a significant loss of genetic diversity from south to north, a strong pattern of allele surfing across nearly all loci, and an increase in genetic differentiation towards the northern range margin. Results from SDMs suggest that southward range retraction during the late glacial period split the distribution into geographically distinct refugia.Main conclusionsThe strong genetic differentiation mirrors the effects of long‐term isolation of P. muralis in multiple refugia. Post‐glacial recolonization of Northern Europe has taken place from two distinct refugia, most probably along river systems (Rhône, Rhine, Moselle) and along the Atlantic coastline, with subsequent nested elimination of genetic diversity and increasing genetic differentiation at the northern range margin.

Setting Conservation Priorities in a Widespread Species: Phylogeographic and Physiological Variation in the Lake Chub, Couesius plumbeus (Pisces: Cyprinidae)

Defining units of conservation below the species level is a widely accepted conservation priority, but is especially challenging for widespread taxa that have experienced diverse geographic histories and exist across heterogenous environments. The lake chub (Pisces: Couesius plumbeus) is a widespread freshwater fish in North America and occurs from the southcentral USA to northwestern Alaska and Canada. We used mtDNA sequence analysis to test for divergent lineages predicted to occur as a result of survival of lake chub in distinct glacial refugia. Lake chub consisted of two major mtDNA lineages separated by 3.8% sequence divergence which are probably late to pre-Pleistocene in origin. We combined these data with those consistent with thermal adaptation in fish living in thermal springs versus those living in a lake with wide seasonal temperature variation, and with data on distribution of lake chub in major watershed units. We assessed these data against objective criteria developed to identify conservation units under Canadian endangered species legislation. Our analysis identified twelve major units of conservation within C. plumbeus that could be assessed under Canada’s Species-at-Risk Act. Our study illustrates how different character traits manifested at very different spatial scales can be used to define conservation units within widely-distributed taxa.

Genetic structure and phylogeography of a European flagship species, the white‐tailed sea eagle Haliaeetus albicilla

We analysed 120 white‐tailed sea eagles Haliaeetus albicilla from eastern (Poland and Estonia) and southeastern (Serbian Danube population) Europe for genetic variability and structuring at the mitochondrial control region and seven nuclear microsatellite loci. We combined this new dataset with sequence and genotype data from previous analyses covering Greenland and Eurasia (total sample sizes of 420 and 186 individuals for mtDNA and microsatellites, respectively) to address the following questions: 1) does the large eastern population in Europe add significantly to the species‘ overall genetic diversity? 2) Do the new sequence data match the clinal distribution pattern (west to east) of the two major mtDNA lineages? 3) Does the preliminary hypothesis of two nuclear genetic clusters recently found in this species hold for the whole of Europe, and do these clusters show a geographic pattern? Our results confirmed Europe as a stronghold of genetic diversity in white‐tailed sea eagles, and the east of the continent contributed disproportionately to this, the reason being the admixture of eagles with different genetic background. As hypothesised, both mitochondrial lineages were recovered also in eastern Europe, but the globally more eastern lineage was dominant. The presence of two microsatellite clusters was also confirmed, and these groups, too, show a non‐random geographic distribution, with, except for Poland, a high proportion of ‘eastern‐type’ eagles in the populations of east–central and eastern Europe.