Molecular systematics and evolution of Regina and the thamnophiine snakes.

Snakes of the tribe Thamnophiini represent an ecologically important component of the herpetofauna in a range of habitats across North America. Thamnophiines are the best-studied colubrids, yet little is known of their systematic relationships. A molecular phylogenetic study of 32 thamnophiine species using three complete mitochondrial genes (cytochrome b, NADH dehydrogenase subunit 2, and 12S ribosomal DNA) recovered a well-supported phylogeny with three major clades: a garter snake group, a water snake group, and a novel semifossorial group. The historically contentious genus Regina, which contains the crayfish-eating snakes, is polyphyletic. The phylogeographic pattern of Thamnophis is consistent with an hypothesis of at least one invasion of northern North America from Mexico.     

Phylogeographic analyses reveal distinct lineages of the liverworts Metzgeria furcata (L.) Dumort. and Metzgeria conjugata Lindb. (Metzgeriaceae) in Europe and North America

Seed plant genera often exhibit intercontinental disjunctions where different species are found on different continents. Many morphologically circumscribed bryophyte species exhibit similar disjunctions. We used nucleotide sequences from the plastid and nuclear genomes to test hypotheses of phylogeography within representatives of the genus Metzgeria: Metzgeria furcata, Metzgeria conjugata, and Metzgeria myriopoda. The first two species have sexual and asexual populations, exhibit disjunctions between North America and Europe, and have been split into separate species, numerous subspecies or varieties. The third species occurs in eastern North America but is not reported from Europe. Phylogenetic analyses resolved three distinct lineages within the morphologically defined species, M. furcata: one in North America, and two in Europe. Similarly, three morphologically cryptic clades of M. conjugata were resolved by the molecular data: northern North America, Europe, and south‐eastern North America. For both species, molecular divergence among taxa occurred in the absence of morphological change. In the case of M. myriopoda, all plants from eastern North America were both morphologically uniform and genetically homogeneous (although not identical). The present study provides significant insight into a plant group with complex taxonomy, and indicates that these liverwort taxa with wide distributions, extreme sex ratios, and continental disjunctions harbor cryptic lineages. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 745–756.     

Phylogenetics and biogeography of eastern Asian–North American disjunct genus Pachysandra (Buxaceae) inferred from nucleotide sequences

Abstract Pachysandra is an eastern Asian–North American disjunct genus with three species, two in eastern Asia (Pachysandra axillaris and Pachysandra terminalis) and one in eastern North America (Pachysandra procumbens). Although morphological and cytological studies suggest a close affinity of P. procumbens with P. axillaris, molecular data from nuclear and chloroplast DNA regions have provided conflicting signals. In this study, we tested previous phylogenetic hypotheses using sequences of nuclear ribosomal DNA internal transcribed spacers and chloroplast ndhF gene from multiple individuals of each of the three species. We also estimated the time of divergence between eastern Asia and eastern North America. Our results support the morphological and cytological conclusion that P. procumbens is more closely related to P. axillaris than to P. terminalis. The estimated time of divergence of P. axillaris and P. procumbens was 14.6±5.5 mya, consistent with estimates from many other eastern Asian–North American disjunct genera. The migration of Pachysandra populations from eastern Asia to North America might have occurred by way of the North Atlantic land bridge.     

Patterns of genetic differentiation in Thamnophis and Taricha from the Pacific Northwest

Aim The co‐evolutionary interaction between the common garter snake, Thamnophis sirtalis, and the rough‐skinned newt, Taricha granulosa, takes place throughout much of the Pacific Northwest (North America). The biogeography of the Pacific Northwest has been heavily influenced by the last Pleistocene glaciation, which reached a maximum as late as 14,000 yr bp . We researched: (1) what type of population structure is present for garter snakes and newts, (2) whether the population structure of these species is consistent with a Pleistocene glaciation hypothesis, and (3) how population structure and migration possibly affect co‐evolution between these species. Location The Pacific Northwest of North America, specifically northern California, Oregon and Washington in the USA. Methods We sampled approximately 20 populations for each species from three different transects. Using microsatellite markers and tissue samples from both species, we quantified the population structure for both species. Individual‐based assignment tests were used to estimate contemporary migration rates. Results Both Th. sirtalis and Ta. granulosa exhibited little genetic differentiation among our study sites, even among those separated by large distances. Significant population structure was detected on multiple geographic scales. Differences in population structure were observed among transects and between garter snake and newt transects. Contemporary migration rate estimates indicate high levels of genetic exchange between populations. Main conclusions Prior to this study, little was known about the fine‐scale population structure of either species in this region. Patterns of population structure for garter snakes and newts reflect a shared biogeographical history affected by the Pleistocene glaciation in the Pacific Northwest. Both species apparently migrate frequently between populations, thus potentially retarding the process of adaptive co‐evolution. We find that populations from a northern coastal transect (Washington) are most likely to be locally adapted.     

One haploid parent contributes 100% of the gene pool for a widespread species in northwest North America

The monoicous peatmoss Sphagnum subnitens has a tripartite distribution that includes disjunct population systems in Europe (including the Azores), northwestern North America and New Zealand. Regional genetic diversity was highest in European S. subnitens but in northwestern North America, a single microsatellite‐based multilocus haploid genotype was detected across 16 sites ranging from Coos County, Oregon, to Kavalga Island in the Western Aleutians (a distance of some 4115 km). Two multilocus haploid genotypes were detected across 14 sites on South Island, New Zealand. The microsatellite‐based regional genetic diversity detected in New Zealand and North American S. subnitens is the lowest reported for any Sphagnum. The low genetic diversity detected in both of these regions most likely resulted from a founder event associated with vegetative propagation and complete selfing, with one founding haploid plant in northwest North America and two in New Zealand. Thus, one plant appears to have contributed 100% of the gene pool for the population systems of S. subnitens occurring in northwest North America, and this is arguably the most genetically uniform group of plants having a widespread distribution yet detected. Although having a distribution spanning 12.5° of latitude and 56° of longitude, there was no evidence of any genetic diversification in S. subnitens in northwest North America. No genetic structure was detected among the three regions, and it appears that European plants of S. subnitens provided the source for New Zealand and northwest North American populations.     

The historical ecology of yellow perch (Perca flavescens[Mitchill]) and their parasites

Aim To determine the origins of the host–parasite association between among yellow perch (Perca flavescens[Mitchill]) and the parasites Crepidostomum cooperi Hopkins, Proteocephalus pearsei La Rue and Urocleidus adspectus Beverly Burton. Of secondary interest are the parasites Bunodera luciopercae (Muller) and Proteocephalus percae (Muller) predictably associated with the Eurasian perch. Location The areas considered are the Holarctic, since the upper‐Cretaceous, and contemporary North America. Methods Published and new information from host and parasite phylogenies, palaeontology, palaeogeography and plate tectonics and host biology is incorporated to assess the origins of yellow perch and several of its parasites. This information is used to determine the origins for these host–parasite associations. Results Cladistic analysis suggests a Laurasian origin for Percidae and Perca, and that Perca is sister to the other genera in the family. Parasite phylogenies support a North American origin for the three species associated with yellow perch and a Laurasian origin for B. luciopercae. Proteocephalus pearsei and P. percae are not sister taxa. The fossil record for Perca dates to the Miocene in Europe and the Pleistocene in North America. North America and Europe were connected across the North Atlantic since at least the upper Cretaceous with separation complete by the Miocene. Europe was separated from Asia by the Obik Sea from the late Cretaceous until the Oligocene. Western cordillera orogeny and its accompanying high rates of water flow and Pleistocene glaciation represent barriers to Perca dispersal. Main conclusions The origin of Perca in North America dates at least to the late Oligocene when North America and Europe were connected across the North Atlantic and Europe and Asia were separate landmasses, and does not result from Pleistocene dispersal across Beringia from Asia. The present disjunction of Perca species in North America and Europe is due to the vicariant separation of North America and Europe. Based on the available information, yellow perch and its parasites have a North America origin. The association between yellow perch and the parasites in all cases is a consequence of host switching from other sympatric host species in North America and is not explained by co‐speciation. Even the association between the host‐specific Urocleidus adspectus and yellow perch originated with a host switch and is not due to co‐speciation. The basis for this host switching is geographical and ecological sympatry, especially shared feeding habits, with other North American fish hosts.     

Isolation and characterization of microsatellite loci in the black rat snake (Elaphe obsoleta)

We obtained molecular markers useful for population level studies of the black rat snake (Elaphe obsoleta) by screening genomic DNA libraries enriched for dinucleotide, tetranucleotide, and pentanucleotide microsatellite repeats. Following sequencing of the positive clones, 11 pairs of primers were designed for polymorphic loci and their variability assessed in > 350 individuals from four populations in North America. The loci had between 9 and 40 alleles and observed heterozygosities ranged from 0.071 to 0.87. Some of these pairs of primers also successfully amplified DNA from two other snake species.     

From Africa via Europe to South America: migrational route of a species‐rich genus of Neotropical lowland rain forest trees (Guatteria,Annonaceae)

Aim Several recent studies have suggested that a substantial portion of today’s plant diversity in the Neotropics has resulted from the dispersal of taxa into that region rather than by vicariance. In general, three routes have been documented for the dispersal of taxa onto the South American continent: (1) via the North Atlantic Land Bridge, (2) via the Bering Land Bridge, or (3) from Africa directly onto the continent. Here a species‐rich genus of Neotropical lowland rain forest trees (Guatteria, Annonaceae) is used as a model to investigate these three hypotheses. Location The Neotropics. Methods The phylogenetic relationships within the long‐branch clade of Annonaceae were reconstructed (using maximum parsimony, maximum likelihood and Bayesian inference) in order to gain insight in the phylogenetic position of Guatteria. Furthermore, Bayesian molecular dating and Bayesian dispersal–vicariance (Bayes‐DIVA) analyses were undertaken. Results Most of the relationships within the long‐branch clade of Annonaceae were reconstructed and had high support. However, the relationship between the Duguetia clade, the Xylopia–Artabotrys clade and Guatteria remained unclear. The stem node age estimate of Guatteria ranged between 49.2 and 51.3 Ma, whereas the crown node age estimate ranged between 11.4 and 17.8 Ma. For the ancestral area of Guatteria and its sister group, the area North America–Africa was reconstructed in 99% of 10,000 DIVA analyses, while South America–North America was found just 1% of the time. Main conclusions The estimated stem to crown node ages of Guatteria in combination with the Bayes‐DIVA analyses imply a scenario congruent with an African origin followed by dispersal across the North Atlantic Land Bridge in the early to middle Eocene and further dispersal into North and Central America (and ultimately South America) in the Miocene. The phylogenetically and morphologically isolated position of the genus is probably due to extinction of the North American and European stem lineages in the Tertiary.     

Phylogenetic and biogeographical relationships of the Sander pikeperches (Percidae: Perciformes): patterns across North America and Eurasia

North America and Eurasia share several closely related taxa that diverged either from the breakup of the Laurasian supercontinent or later closures of land bridges. Their modern population structures were shaped in Pleistocene glacial refugia and via later expansion patterns, which are continuing. The pikeperch genus Sander contains five species – two in North America (S. canadensis and S. vitreus) and three in Eurasia (S. lucioperca, S. marinus, and S. volgensis) – whose evolutionary relationships and relative genetic diversities were previously unresolved, despite their fishery importance. This is the first analysis to include the enigmatic and rare sea pikeperch S. marinus, nuclear DNA sequences, and multiple mitochondrial DNA regions. Bayesian and maximum‐likelihood trees from three mitochondrial and three nuclear gene regions support the hypothesis that Sander diverged from its sister group Romanichthys/Zingel ～24.6 Mya. North American and Eurasian Sander then differentiated ～20.8 Mya, with the former diverging ～15.4 Mya, congruent with North American fossils dating to ～16.3–13.6 Mya. Modern Eurasian species date to ～13.8 Mya, with S. volgensis being basal and comprising the sister group to S. lucioperca and S. marinus, which diverged ～9.1 Mya. Genetic diversities of the North American species are higher than those in Eurasia, suggesting fewer Pleistocene glaciation bottlenecks. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 156–179.     

Beneath the bark: associations among Sirex noctilio development,bluestain fungi,and pine host species in North America

1. Invasive species with global distributions encounter unique environmental and biotic variables that can greatly affect the magnitude of their impact. The European woodwasp, Sirex noctilio Fabricius, is a prime example that has invaded climatically and ecologically distinct ecosystems across the Southern Hemisphere and, more recently, North America. 2. Northeastern North America presents a unique set of conditions, including pine host species, native parasitoids, a diverse assemblage of native co‐colonising insects, and fungal associates of these co‐colonisers. In North America, S. noctilio attacks both a naturalised ancestral host (Pinus sylvestris L.) from Europe and a naïve native host (P. resinosa). A large assemblage of insects and their associated bluestaining ophiostomatoid fungi colonise these pines. Competition between S. noctilio and this group is a hypothesised mechanism of biotic resistance in the invaded region of North America, possibly via superior resource capture abilities or alteration of host tissue by bluestain fungi. 3. Investigating these ecological interactions is challenging because they manifest deep in the xylem tissue. To overcome this, 30 experimentally stressed trees were systematically dissected with an electric log splitter to investigate the effects of bluestain fungi and tree factors on S. noctilio development and parasitism by native hymenopterans. 4. Body size and colonisation density were affected by pine species, with S. noctilio being 25% larger and densities three‐fold greater in P. sylvestris than P. resinosa. Survivorship was slightly negatively correlated with the proportional volume of bluestain infection. Interestingly, rhyssine (Rhyssa and Megarhyssa spp.) parasitism responded positively to greater S. noctilio density, but there was no density relationship with Ibalia leucospoides ensiger parasitism. Pine host species appears to play a strong role in S. noctilio development, which is important considering uninvaded regions of North America have a diversity of pine species that likely vary in their susceptibility to this woodborer.     

Phylogenetic assessment and biogeographic analyses of tribe <Emphasis Type="Italic">Peracarpeae</Emphasis> (Campanulaceae)

Peracarpeae is a small tribe consisting of three genera: Homocodon, Heterocodon and Peracarpa, with a disjunct distribution between eastern Asia and western North America. Homocodon is endemic to southwestern China and was previously placed in the western North American genus Heterocodon. Our phylogenetic analysis using four plastid markers (matK, atpB, rbcL and trnL-F) suggests the polyphyly of Peracarpeae. Homocodon is sister to a clade consisting of the eastern Asian Adenophora, Hanabusaya and species of Campanula from the Mediterranean region and North America, rather than forming a clade with Heterocodon. Homocodon and its Eurasia relatives are estimated to have diverged in the early Miocene (16.84 mya, 95% HPD 13.35–21.45 mya). The eastern Asian Peracarpa constitutes a clade with the North American Heterocodon, Githopsis and three species of Campanula, supporting a disjunction between eastern Asia and North America in Campanulaceae. The Asian-North American disjunct lineages diverged in the early Miocene (16.17 mya, 95% HPD 13.12–20.9 mya). The biogeographic analyses suggest that Homocodon may be a relict of an early radiation in eastern Asia, and that Peracarpa and its closest North American relatives most likely originated from a Eurasian ancestor.     

Demographic expansion and genetic load of the halophyte model plant Eutrema salsugineum

The halophyte model plant Eutrema salsugineum (Brassicaceae) disjunctly occurs in temperate to subarctic Asia and North America. This vast, yet extremely discontinuous distribution constitutes an ideal system to examine long‐distance dispersal and the ensuing accumulation of deleterious mutations as expected in expanding populations of selfing plants. In this study, we resequenced individuals from 23 populations across the range of E. salsugineum. Our population genomic data indicate that E. salsugineum migrated “out of the Altai region” at least three times to colonize northern China, northeast Russia and western China. It then expanded its distribution into North America independently from northeast Russia and northern China, respectively. The species colonized northern China around 33.7 thousand years ago (kya) and underwent a considerable expansion in range size approximately 7–8 kya. The western China lineage is likely a hybrid derivative of the northern China and Altai lineages, originating approximately 25–30 kya. Deleterious alleles accumulated in a stepwise manner from (a) Altai to northern China and North America and (b) Altai to northeast Russia and North America. In summary, E. salsugineum dispersed from Asia to North America and deleterious mutations accumulated in a stepwise manner during the expansion of the species’ distribution.     

Phylogeny and Phytogeography of Eupatorium (Eupatorieae, Asteraceae): Insights from Sequence Data of the nrDNA ITS Regions and cpDNA RFLP

Eupatorium were examined by sequencing the internal transcribed spacers (ITS) of nuclear ribosomal DNA and restriction site analysis of chloroplast DNA. Molecular data provided strong evidence that (1) this genus originated in North America, (2) the genus diverged into three morphological species groups, Eutrochium, Traganthes and Uncasia in North America, and (3) one of the North American Uncasia lineages migrated into temperate Europe and eastern Asia over the Bering land bridge. The estimated divergence times support a late Miocene to early Pliocene migration from North America to Eurasia via the Bering land bridge. A European species was sister to all of the eastern Asian species examined. The disjunct distribution pattern of the genus Eupatorium is incongruent with the classical Arcto-Tertiary geoflora concept. Received 13 September 1999/ Accepted in revised form 4 January 2000     

A molecular phylogeny of the <Emphasis Type="Italic">Lecanora varia</Emphasis> group,including a new species from western North America

The phylogeny of the Lecanora varia group is inferred from ITS sequences using Bayesian and ML phylogenetic analysis methods. According to our data, usnic acid-containing, corticolous Lecanora species do not form a monophyletic group but occur in three major clades together with other groups of Lecanora and Protoparmeliopsis. The new combination Lecanora filamentosa (Stirt.) Elix & Palice is proposed. The new species Lecanora schizochromatica is described from northwestern North America. It produces atranorin as a major substance and is closely related to the morphologically and chemically similar L. filamentosa. The American Biatora pullula Tuck. is synonymised with Lecanora anopta Nyl., which is hereby reported for continental North America for the first time. The phylogenetic relationships between the major clades of Lecanora are still largely unresolved and require more intensive taxon and character sampling.     

CHROMOSOME NUMBERS AND EVOLUTION IN NORTH AMERICAN SPECIES OF LINUM

Among the North American species of Linum there are three basic chromosome numbers representing invasions from the Old World of three distinct evolutionary lines. N = 9 is found only in the blue-flowered group represented in North America by two species, L. lewisii and L. pratense which are closely related to and may be conspecific with the Old World L. perenne. The basic number for the yellow-flowered species is n = 18 which is characteristic of the Scabrella and Virginiana subgroups. The loss of chromosomes in the Neo-mexicana (n = 13) and Sulcata-Rigida (n = 15) subgroups suggests that the basic haploid number of 18 might be a polyploid derivative of an Old World ancestor with n = 9. The incidence of n = 9 among Old World species of Linum may indicate that this represents an ancestral condition. Linum catharticum has n = 8; this number and features of morphology and distribution suggest that it is not directly related to either the blue-flowered or yellow-flowered groups in North America but represents a separate introduction on this continent.     

The population genetics of the causative agent of snake fungal disease indicate recent introductions to the USA

Snake fungal disease (SFD; ophidiomycosis), caused by the pathogen Ophidiomyces ophiodiicola (Oo), has been documented in wild snakes in North America and Eurasia, and is considered an emerging disease in the eastern United States of America. However, a lack of historical disease data has made it challenging to determine whether Oo is a recent arrival to the USA or whether SFD emergence is due to other factors. Here, we examined the genomes of 82 Oo strains to determine the pathogen’s history in the eastern USA. Oo strains from the USA formed a clade (Clade II) distinct from European strains (Clade I), and molecular dating indicated that these clades diverged too recently (approximately 2,000 years ago) for transcontinental dispersal of Oo to have occurred via natural snake movements across Beringia. A lack of nonrecombinant intermediates between clonal lineages in Clade II indicates that Oo has actually been introduced multiple times to North America from an unsampled source population, and molecular dating indicates that several of these introductions occurred within the last few hundred years. Molecular dating also indicated that the most common Clade II clonal lineages have expanded recently in the USA, with time of most recent common ancestor mean estimates ranging from 1985 to 2007 CE. The presence of Clade II in captive snakes worldwide demonstrates a potential mechanism of introduction and highlights that additional incursions are likely unless action is taken to reduce the risk of pathogen translocation and spillover into wild snake populations.

Snake fungal disease is an emerging disease in eastern North America, but the origins of the disease have been unclear. This study uses population genetic data to show that the fungus that causes the disease was introduced multiple times to North America over the last few hundred years.     

Comparative phylogeography of Amphicarpaea legumes and their root-nodule symbionts in Japan and North America

Aim Relationships of eastern Asian and eastern North American populations of legumes in the genus Amphicarpaea Elliot ex. Nuttall (Phaseoleae–Glycininae) and their root nodule bacteria (Bradyrhizobium Jordan) were analysed to test whether both organisms share an identical biogeographic history. Location Japan and eastern North America (New York and Illinois). Methods Sequences of three plant genes (chloroplast trnL region, nuclear ribosomal ITS, and histone H3‐D) and a segment of the bacterial ribosomal region (partial 16S rRNA and 23S rRNA genes, and the 16S rRNA–23S rRNA ITS) were used to analyse phylogenetic relationships. Results For plants, Japanese populations formed a sister group to a well‐supported clade of all North American genotypes. For nodule bacteria associated with Amphicarpaea, isolates from North America did not form a single clade relative to Asian genotypes. Japanese Bradyrhizobium isolates were closely related to particular sub‐groups of North American bacteria (lineages ‘B’ and ‘C’), with other American bacteria branching earlier. Main conclusions Plants and bacteria showed clear deviations from a pattern of parallel cladogenesis. The most basal Amphicarpaea lineage was associated with a recently‐diverged bacterial group, while one recently‐diverged plant lineage had symbionts that branched in a basal position relative to the other Amphicarpaea bacteria. When analysed with data on symbiotic compatibility from inoculation experiments, the molecular phylogenies suggested that for plants, at least one transition has occurred toward more promiscuous nodulation behaviour. Among bacteria, strains with narrow host range on Amphicarpaea appear to be ancestral to symbiotic generalists.     

Des serpents (Reptilia,Squamata) de type nord-américain dans le Miocène Français. Évolution parallèle ou dispersion?

The French Miocene (Orleanian and Astaracian) yielded five new species of snakes referable to four genera from the Neogene or Recent of North America: Texasophis meini nov. sp., Paleoheterodon arcuatus nov. sp., Neonatrix europaea nov. sp., Neonatrix crassa nov. sp. (Colibridae) and Micrurus gallicus nov. sp. («Elapidae). It is suggested that Micrurus and the previously described boid snake Albaneryx from the French Miocene probably represent lineages that originated in North America and reached Europe by way of Asia. Paleoheterodon and Neonatrix could have originated in Asia and subsequently reached Europe on one hand and North America on the other. Texasophis may have originated in North America and followed the same route as Micrurus and Albaneryx, but an alternative hypothesis is that it originated in Asia and spread toward Europe and North America in the same manner as Paleoheterodon and Neonatrix.     

Systematics and macroevolution of extant and fossil scalopine moles (Mammalia,Talpidae)

Scalopini is one of the two fully fossorial mole tribes in the family Talpidae, with remarkable adaptations to subterranean lifestyles. Most living Scalopini species are distributed in North America while a sole species occurs in China. On the other hand, scalopine fossils are found in both Eurasia and North America from upper Oligocene strata onwards, implying a complex biogeographical history. The systematic relationships of both extant and fossil Scalopini across North America and Eurasia are revised by conducting phylogenetic analyses using a comprehensive morphological character matrix together with 2D geometric–morphometric analyses of the humeral shape, with a specific emphasis on Mioscalops, a genus commonly found in North America and formerly known as Scalopoides. Our phylogenetic analyses support the monophyly of the tribe Scalopini as well as a proposed two‐subtribe‐division scenario of Scalopini (i.e. Scalopina and Parascalopina), although Proscapanus could not be assigned to either subgenus. Our geometric–morphometric analyses indicate that the European Mioscalops from southern Germany should be allocated to Leptoscaptor, which in turn implies that Mioscalops may be endemic to North America and never arrived in Europe. Examination of biogeographical patterns does not unambiguously determine the geographical origin of Scalopini. Nevertheless, it does support multiple transcontinental colonization events across Asia, Europe and North America. Scapanulus oweni, distributed in central China, is the only remaining representative of one of those out‐of‐North‐America migrations, whereas scalopine moles are common in North America nowadays with up to five species.     

Extinct Beringian wolf morphotype found in the continental U.S. has implications for wolf migration and evolution

Pleistocene diversity was much higher than today, for example there were three distinct wolf morphotypes (dire, gray, Beringian) in North America versus one today (gray). Previous fossil evidence suggested that these three groups overlapped ecologically, but split the landscape geographically. The Natural Trap Cave (NTC) fossil site in Wyoming, USA is an ideally placed late Pleistocene site to study the geographical movement of species from northern to middle North America before, during, and after the last glacial maximum. Until now, it has been unclear what type of wolf was present at NTC. We analyzed morphometrics of three wolf groups (dire, extant North American gray, Alaskan Beringian) to determine which wolves were present at NTC and what this indicates about wolf diversity and migration in Pleistocene North America. Results show NTC wolves group with Alaskan Beringian wolves. This provides the first morphological evidence for Beringian wolves in mid‐continental North America. Their location at NTC and their radiocarbon ages suggest that they followed a temporary channel through the glaciers. Results suggest high levels of competition and diversity in Pleistocene North American wolves. The presence of mid‐continental Beringian morphotypes adds important data for untangling the history of immigration and evolution of Canis in North America.