Mitochondrial cytochrome b sequence yields new insight into the speciation of social voles in south-west Asia

We established a cytochrome b (cyt b ) phylogeny for six species of social voles. A Bayesian approach to phylogenetic reconstruction (BI) and a maximum likelihood (ML) tree revealed a dichotomy into two major clusters, namely a Microtus guentheri cluster and a M. socialis cluster. The three main lineages that emerged within each of these two clusters were separated by the K2P divergences which are above the intraspecific variation in Microtus . All six species were also retrieved in the minimum spanning network. Within its present taxonomic scope, M. guentheri is paraphyletic and consists of two allopatric sibling species: M. guentheri (Syria, Israel) and M. hartingi (Anatolia and the Balkans). The closest relative to these two species is M. dogramacii , which is possibly a sister species to M. hartingi . The two geographic samples were identified as M. irani , one from Shiraz (Iran) and the other from Balkusan (Turkey). The cyt b sequence confirmed the specific status of M. anatolicus within the M. socialis cluster. Although five species of social voles occur within a radius of < 500 km in the north-eastern corner of the Mediterranean, small-scale sympatry is exceptional. Species richness in this region possibly originates from past fragmentation with subsequent allopatric speciation in refugial areas.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 121–128.     

Holarctic phylogeography of the root vole (Microtus oeconomus): implications for late Quaternary biogeography of high latitudes

A species-wide phylogeographical study of the root vole (Microtus oeconomus) was performed using the whole 1140 base pair mitochondrial (mt) cytochrome b gene. We examined 83 specimens from 52 localities resulting in 65 unique haplotypes. Our results demonstrate that the root vole is divided into four main mtDNA phylogenetic lineages that seem to have largely allopatric distributions. Net divergence estimates (2.0-3.5%) between phylogroups, as well as relatively high nucleotide diversity estimates within phylogroups, indicate that the distinct phylogeographical structure was initiated by historical events that predated the latest glaciation. European root voles are divided into a Northern and a Central mtDNA phylogroup. The mtDNA data in concert with fossil records imply that root voles remained north of the classical refugial areas in southern Europe during the last glacial period. The currently fragmented populations in central Europe belong to a single mtDNA phylogroup. The Central Asian and the North European lineages are separated by the Ural Mountains, a phylogeographical split also found in collared lemmings (Dicrostonyx) and the common vole (M. arvalis). The Beringian lineage occurs from eastern Russia through Alaska to northwestern Canada. This distribution is congruent with the traditional boundaries of the Beringian refugium and with phylogeographical work on other organisms. In conclusion, similarities between the phylogeographical patterns in the root vole and other rodents, such as Arctic and subarctic lemmings, as well as more temperate vole species, indicate that late Quaternary geological and climatic events played a strong role in structuring northern biotic communities.     

Phylogeography of the common vole (Microtus arvalis) with particular emphasis on the colonization of the Orkney archipelago

To investigate the human introduction of the common vole Microtus arvalis onto the Orkney islands, the complete cytochrome b gene was sequenced in 41 specimens from both Orkney (four localities) and elsewhere in their range (26 localities). Orkney voles belonged to the same phylogenetic lineage, 'Western', as individuals from France and Spain indicating southwestern Europe as the most likely source area for the islands. This result is of interest with respect to the movement and trading links of the Neolithic people who likely transported the voles. As well as the Western lineage, our phylogenetic trees revealed three other purely European lineages: the 'Italian' (single specimen from N. Italy), the 'Central' (Germany, Netherlands, Denmark) and the 'Eastern' (Hungary, Slovakia, Poland, Ukraine, Finland, European Russia). Individuals from European Russia, W. Siberia, Georgia, Ukraine and Armenia formed a fifth distinct lineage coinciding with the distribution of the 'obscurus' chromosomal form of M. arvalis. These phylogeographical data suggest that M. arvalis occupied multiple refugia during the last glaciation.     

Low mitochondrial divergence indicates a rapid expansion across Europe in the weather loach, Misgurnus fossilis (L.)

Several phylogeographic studies using mtDNA sequence data have revealed an expressed geographic structure in nearly every European freshwater fish species studied. The authors present a phylogeographic study of Misgurnus fossilis on the base of 43 specimens from 17 localities across a major part of the known distribution area of M. fossilis . Despite the large geographic distance between the sampling points and their origin from different major European river systems, only eight closely related haplotypes in the sequences of the whole mitochondrial cytochrome b were detected. The most common haplotype I included more than 60% of specimens and occurred in the North Sea basin in northern Germany, in the Danube and Elbe basins in the Czech Republic, in the Nieman basin in Poland and in the Dniester and Vistula basins in the Ukraine. Since the highest number of haplotypes (six out of eight) and the most divergent haplotypes were found in the Danube, the authors tentatively consider the Danube to have acted as a refuge area for Misgurnus during the glaciation maxima in the Pleistocene. From this refuge, the species presumably recolonized Central and Eastern Europe but failed to stretch to Western Europe.     

The complete mitochondrial genome of Microtus fortis calamorum (Arvicolinae, Rodentia) and its phylogenetic analysis

Microtus fortis is a special resource of rodent in China. It is a promising experimental animal model for the study on the mechanism of Schistosome japonicum resistance. The first complete mitochondrial genome sequence for Microtus fortis calamorum, a subspecies of M. fortis (Arvicolinae, Rodentia), was reported in this study. The mitochondrial genome sequence of M. f. calamorum (Genbank: JF261175) showed a typical vertebrate pattern with 13 protein coding genes, 2 ribosomal RNAs, 22 transfer RNAs and one major noncoding region (CR region).The extended termination associated sequences (ETAS-1 and ETAS-2) and conserved sequence block 1 (CSB-1) were found in the CR region. The putative origin of replication for the light strand (O(L)) of M. f. calamorum was 35bp long and showed high conservation in stem and adjacent sequences, but the difference existed in the loop region among three species of genus Microtus. In order to investigate the phylogenetic position of M. f. calamorum, the phylogenetic trees (Maximum likelihood and Bayesian methods) were constructed based on 12 protein-coding genes (except for ND6 gene) on H strand from 16 rodent species. M. f. calamorum was classified into genus Microtus, Arvcicolinae for the highly phylogenetic relationship with Microtus kikuchii (Taiwan vole). Further phylogenetic analysis results based on the cytochrome b gene ranged from M. f. calamorum to one of the subspecies of M. fortis, which formed a sister group of Microtus middendorfii in the genus Microtus.     

Historical DNA reveals the phylogenetic position of the extinct Alpine lynx

During the last two centuries, lynx populations have undergone severe declines and extinctions in Europe. The Alpine lynx, once distributed across the whole Alpine arc, became extinct due to direct human prosecution and deprivation of its main prey in the 1930s. Similar to the Iberian lynx Lynx pardinus , its taxonomy has been subject to several controversies. Moreover, knowing the taxonomic status of the Alpine lynx will help to define conservation units of extant lynx populations in Europe. In this study, we investigated two mitochondrial DNA regions in museum specimens ( n =15) representing the autochthonous Alpine population and in samples from extant Eurasian lynx Lynx lynx populations in Europe and Asia ( n =17). Phylogenetic analysis (cytochrome b , 345 bp) placed the Alpine lynx within the Eurasian lynx lineage. Among all individuals examined, seven different haplotypes (control region, 300 bp) were observed but no unique Alpine haplotype was discovered. Haplotypes of the extinct Alpine population were identical to previously described haplotypes in Scandinavian lynx signifying a recent genetic ancestry with current European populations. Moreover, our genetic data suggest two distinct glacial refugia for the Carpathian and Balkan population. Overall this study demonstrates that historical DNA from extinct populations can help to disentangle the phylogenetic relationships and historical biogeography of taxa with only a limited number of extant populations remaining.     

Out of Africa? Phylogenetic relationships between Falco biarmicus and the other hierofalcons (Aves: Falconidae)

The phylogeographic history of the lanner falcon ( Falco biarmicus ) and the phylogenetic relationships among hierofalcons ( F. biarmicus , Falco cherrug , Falco jugger and Falco rusticolus ) were investigated using mitochondrial (mt) DNA sequences. Of the two non-coding mt sections tested, the control region (CR) appeared more suitable as phylogenetic marker sequence compared with the pseudo control region (ΨCR). For the comprehensive analysis samples from a broad geographic range representing all four hierofalcon species and their currently recognized subspecies were included. Moreover, samples of Falco mexicanus were analysed to elucidate its phylogenetic relationships to the hierofalcons. The sequence data indicate that this species is more closely related to Falco peregrinus than to the hierofalcons. In the DNA-based trees and in the maximum parsimony network all hierofalcons appear closely related and none of the species represents a monophyletic group. The close relationships among haplotypes suggest that the hierofalcon complex is an assemblage of morphospecies not yet differentiated in the genetic markers used in the present study and that the radiation of the four hierofalcon species took place rather recently. Based on the high intraspecific diversity found within F. biarmicus we assume an African origin of the hierofalcon complex. The observed pattern of haplotype distribution in the extant species may be due to incomplete lineage sorting of ancestral polymorphisms, and interspecific gene flow through hybridization.     

中国地鼠线粒体DNA 16S rRNA基因序列分析及分子系统发育研究

目的通过克隆分析中国地鼠16S基因的部分序列,对中国地鼠16S基因的结构和功能进行初步探索和揭示。方法从GenBank中已报道的啮齿动物16S基因保守区设计一对引物,进行PCR扩增,测序。用Blastn与GenBank中七种啮齿类动物的16S基因进行序列比较,分析其碱基组成及变异情况,并用邻接法(NJ)、非加权组平均法(UPGMA)构建分子系统树,在分子水平上探讨中国地鼠和其他啮齿类动物的进化关系,对中国地鼠的种属地位进行了进一步验证。结果获得了中国地鼠线粒体16S基因的部分序列,其碱基组成A、T、C、G分别为40.5%、24.5%、18.7%、16.3%,与其他七种啮齿类动物的碱基含量相比,各碱基含量基本相似。NJ进化树表明,中国地鼠、金黄地鼠与欧洲仓鼠先聚为一支,小鼠与大鼠先聚为一支,东方田鼠、台湾田鼠与东欧田鼠先聚为一支。结论中国地鼠和金黄地鼠的亲缘关系最近,与传统的分类地位基本吻合。     

BIODIVERSITY RESEARCH: Genetic diversity in two introduced biofouling amphipods (Ampithoe valida & Jassa marmorata) along the Pacific North American coast: investigation into molecular identification and cryptic diversity

Aim We investigated patterns of genetic diversity among invasive populations of Ampithoe valida and Jassa marmorata from the Pacific North American coast to assess the accuracy of morphological identification and determine whether or not cryptic diversity and multiple introductions contribute to the contemporary distribution of these species in the region. Location Native range: Atlantic North American coast; Invaded range: Pacific North American coast. Methods We assessed indices of genetic diversity based on DNA sequence data from the mitochondrial cytochrome c oxidase subunit I (COI) gene, determined the distribution of COI haplotypes among populations in both the invasive and putative native ranges of A. valida and J. marmorata and reconstructed phylogenetic relationships among COI haplotypes using both maximum parsimony and Bayesian approaches. Results Phylogenetic inference indicates that inaccurate species‐level identifications by morphological criteria are common among Jassa specimens. In addition, our data reveal the presence of three well supported but previously unrecognized clades of A. valida among specimens in the north‐eastern Pacific. Different species of Jassa and different genetic lineages of Ampithoe exhibit striking disparity in geographic distribution across the region as well as substantial differences in genetic diversity indices. Main conclusions Molecular genetic methods greatly improve the accuracy and resolution of identifications for invasive benthic marine amphipods at the species level and below. Our data suggest that multiple cryptic introductions of Ampithoe have occurred in the north‐eastern Pacific and highlight uncertainty regarding the origin and invasion histories of both Jassa and Ampithoe species. Additional morphological and genetic analyses are necessary to clarify the taxonomy and native biogeography of both amphipod genera.     

Eastern Beringian biogeography: historical and spatial genetic structure of singing voles in Alaska

Aim  Pleistocene climatic cycles have left marked signatures in the spatial and historical genetic structure of high‐latitude organisms. We examine the mitochondrial (cytochrome b) genetic structure of the singing vole, Microtus miurus (Rodentia: Cricetidae: Arvicolinae), a member of the Pleistocene Beringian fauna, and of the insular vole, Microtus abbreviatus, its putative sister species found only on the St Matthew Archipelago. We reconstruct the phylogenetic and phylogeographical structure of these taxa, characterize their geographical partitioning and date coalescent and cladogenetic events in these species. Finally, we compare the recovered results with the phylogenetic, coalescent and spatial genetic patterns of other eastern Beringian mammals and high‐latitude arvicoline rodents. Location  Continental Alaska (alpine and arctic tundra) and the St Matthew Archipelago (Bering Sea). Methods  We generated and analysed cytochrome b sequences of 97 singing and insular voles (M. miurus and M. abbreviatus) from Alaska. Deep evolutionary structure was inferred by phylogenetic analysis using parsimony, maximum likelihood and Bayesian approaches; the geographical structure of genetic diversity was assessed using analysis of molecular variance and network analysis; ages of cladogenetic and coalescent events were estimated using a relaxed molecular clock model with Bayesian approximation. Results  Regional nucleotide diversity in singing voles is higher than in other high‐latitude arvicoline species, but intra‐population diversity is within the observed range of values for arvicolines. Microtus abbreviatus specimens are phylogenetically nested within M. miurus. Molecular divergence date estimates indicate that current genetic diversity was formed in the last glacial (Wisconsinan) and previous interglacial (Sangamonian) periods, with the exception of a Middle Pleistocene split found between samples collected in the Wrangell Mountains region and all other singing vole samples. Main conclusions  High levels of phylogenetic and spatial structure are observed among analysed populations. This pattern is consistent with that expected for a taxon with a long history in Beringia. The spatial genetic structure of continental singing voles differs in its northern and southern ranges, possibly reflecting differences in habitat distribution between arctic and alpine tundra. Our phylogenetic results support the taxonomic inclusion of M. miurus in its senior synonym, M. abbreviatus.     

Integrative taxonomy of the Italian pine voles,Microtus savii group (Cricetidae,Arvicolinae)

The Italian endemic voles belonging to the Microtus savii group (de Sélys‐Longchamps, 1838) include five controversial taxa, namely M. savii savii, M. s. niethammericus Contoli 2003 ; M. s. tolfetanus Contoli 2003 , M. s. nebrodensis (Minà‐Palumbo, 1868) and M. brachycercus (Lehamann, 1961). A previous mtDNA phylogenetic analysis revealed three main mtDNA lineages that do not correspond to the recognized taxa. Here, we perform a comprehensive univariate and multivariate analysis of craniodental measurements in 151 specimens from 54 geographic samples belonging to the three main phylogroups. Qualitative data on the skull of 126 specimens were also obtained from 54 localities. Moreover, we sequenced the nuclear DNA β‐fibrinogen intron 7 (β‐fibint7) of 23 specimens from 12 localities and Cyt b sequences of 11 new specimens from seven localities. The pattern of morphological and cytogenomic divergence does not corroborate the current taxonomic framework and indicates that M. nebrodensis is a Sicilian endemic species.     

Positioning the Red Deer (Cervus elaphus) Hunted by the Tyrolean Iceman into a Mitochondrial DNA Phylogeny

In the last years several phylogeographic studies of both extant and extinct red deer populations have been conducted. Three distinct mitochondrial lineages (western, eastern and North-African/Sardinian) have been identified reflecting different glacial refugia and postglacial recolonisation processes. However, little is known about the genetics of the Alpine populations and no mitochondrial DNA sequences from Alpine archaeological specimens are available. Here we provide the first mitochondrial sequences of an Alpine Copper Age Cervus elaphus. DNA was extracted from hair shafts which were part of the remains of the clothes of the glacier mummy known as the Tyrolean Iceman or Ötzi (5,350–5,100 years before present). A 2,297 base pairs long fragment was sequenced using a mixed sequencing procedure based on PCR amplifications and 454 sequencing of pooled amplification products. We analyzed the phylogenetic relationships of the Alpine Copper Age red deer''s haplotype with haplotypes of modern and ancient European red deer. The phylogenetic analyses showed that the haplotype of the Alpine Copper Age red deer falls within the western European mitochondrial lineage in contrast with the current populations from the Italian Alps belonging to the eastern lineage. We also discussed the phylogenetic relationships of the Alpine Copper Age red deer with the populations from Mesola Wood (northern Italy) and Sardinia.     

Phylogeny of the Mexican coastal leopard frogs of the Rana berlandieri group based on mtDNA sequences

Phylogenetic relationships among specimens from 25 different locations for the six Mexican coastal leopard frog species of the Rana berlandieri species group were investigated using 797 bp of the mitochondrial 12S rDNA gene. Relationships among the haplotypes obtained were recovered using maximum parsimony and Bayesian analyses. Most of the clades recovered by both tree building methods are strongly supported, but conflicting clades recovered by each analysis are generally poorly supported. Both analyses reject the previously proposed subgroupings of the R. berlandieri species group. Based on the strongly supported relationships, genetic differentiation, and geographic distribution of the haplotypes examined, nine independent lineages appear to comprise the group of study. However, confirmation of the new proposed lineages will require further analyses based on other genetic markers and additional samples that cover their entire geographic distribution. Concordance was noted between Miocene-Pliocene geological and climatic events in Mexico and the relationships recovered among the lineages proposed and their geographic distribution.     

A highly divergent mitochondrial DNA lineage of Microtus agrestis in southern Europe

The Mediterranean peninsulas constitute important areas for endemism and intraspecific variation, and are likely places for cryptic biodiversity. We assessed the phylogeographic pattern of field voles (Microtus agrestis) in southern and central Europe by sequence analysis of a 385-bp fragment of the mitochondrial cytochrome b gene in 74 specimens from 44 localities. The majority of samples consisted of skulls collected from owl pellets. The data revealed a highly distinct cytochrome b lineage in an area ranging from Portugal to Hungary. This southern field vole phylogroup differed by a sequence divergence of 5.6-7.1% from the remaining haplotypes, a level of divergence comparable to that found between known Microtus sibling species. However, this ancient phylogeographic break that dates back many glacial cycles has not been recognised previously by either morphology or karyotype. The southern cytochrome b lineage was further divided into two well-defined sublineages that appear to have derived from different glacial refugia in the Iberian Peninsula.     

Testing species boundaries in an ancient species complex with deep phylogeographic history: genus Xantusia (Squamata: Xantusiidae)

Identification of species in natural populations has recently received increased attention with a number of investigators proposing rigorous methods for species delimitation. Morphologically conservative species (or species complexes) with deep phylogenetic histories (and limited gene flow) are likely to pose particular problems when attempting to delimit species, yet this is crucial to comparative studies of the geography of speciation. We apply two methods of species delimitation to an ancient group of lizards (genus Xantusia) that occur throughout southwestern North America. Mitochondrial cytochrome b and nicotinamide adenine dinucleotide dehydrogenase subunit 4 gene sequences were generated from samples taken throughout the geographic range of Xantusia. Maximum likelihood, Bayesian, and nested cladogram analyses were used to estimate relationships among haplotypes and to infer evolutionary processes. We found multiple well-supported independent lineages within Xantusia, for which there is considerable discordance with the currently recognized taxonomy. High levels of sequence divergence (21.3%) suggest that the pattern in Xantusia may predate the vicariant events usually hypothesized for the fauna of the Baja California peninsula, and the existence of deeply divergent clades (18.8%-26.9%) elsewhere in the complex indicates the occurrence of ancient sundering events whose genetic signatures were not erased by the late Wisconsin vegetation changes. We present a revised taxonomic arrangement for this genus consistent with the distinct mtDNA lineages and discuss the phylogeographic history of this genus as a model system for studies of speciation in North American deserts.     

Phylogeography and ecological niche modelling of the New Zealand stick insect Clitarchus hookeri (White) support survival in multiple coastal refugia

Aim Increasing our understanding of the effects of the Last Glacial Maximum (LGM) and determining the location of refugia requires studies on widely distributed species with dense sampling of populations. We have reconstructed the biogeographic history of Clitarchus hookeri (White), a widespread species of New Zealand stick insect that exhibits geographic parthenogenesis, using phylogeographic analysis and ecological niche modelling. Location New Zealand. Methods We used DNA sequence data from the mitochondrial cytochrome c oxidase subunit I gene to reconstruct phylogenetic relationships among haplotypes from C. hookeri and two undescribed Clitarchus species. We also used distribution data from our own field surveys and museum records to reconstruct the geographic distribution of C. hookeri during the present and the LGM, using ecological niche modelling. Results The ecological niche models showed that the geographic distribution of C. hookeri has expanded dramatically since the LGM. Our model predicted large areas of suitable LGM habitat in upper North Island, and small patches along the east coast of South Island. The phylogeographic analysis shows that populations in the northern half of North Island contain much higher levels of genetic variation than those from southern North Island and South Island, and is congruent with the ecological niche model. The distribution of bisexual populations is also non-random, with males completely absent from South Island and very rare in southern North Island. Main conclusions During the LGM C. hookeri was most likely restricted to several refugia in upper North Island and one or more smaller refugia along the east coast of South Island. The unisexual populations predominate in post-glacial landscapes and are clearly favoured in the recolonization of such areas. Our study exemplifies the utility of integrating ecological niche modelling and phylogeographic analysis.     

Taxonomy, phylogeny, and distribution of Puccinia graminis, the black stem rust: new insights based on rDNA sequence data

Puccinia graminis (Uredinales) is an economically important and common host-alternating rust species on Berberidaceae/Poaceae (subfamilies Pooideae and Panicoideae) that has been spread globally by human activities from an unknown center of origin. To evaluate the taxonomic implications, phylogenetic relationships, and distribution/spread of this complex species, we sequenced and cladistically analyzed the ITS1, 5.8S, and ITS2 regions from herbarium specimens on various host plants from Iran (17), Europe (1), and North America (4). The ITS region plus the 5.8S gene ranged from 686 to 701 bp, including the flanking partial sequences of the 18S and 28S rDNA. Our phylogenetic analysis included 54 bp of the 18S sequence, the entire ITS1 + 5.8S + ITS2, and 58 bp of the 28S sequence. A second analysis used only the last 42 bp of ITS1, and all the 5.8S and ITS2, to incorporate data from additional sequences downloaded from GenBank. In addition to variation in sequence length, there was variation in sequence content. The analysis does not support classical morphology-based taxonomic concepts of the P. graminis complex. Also, host range, host taxonomy, and geographic origin provide minor information on taxonomic relationships. Puccinia graminis is most probably monophyletic. Coevolutionary aspects can hardly be discussed because of lack of sequence data from alternate host specimens. The occurrence of unrelated fungal taxa on the same host species suggests that, besides coevolution with the host, host jumps and hybridization may have played an important role in the evolution of P. graminis. From rDNA data we conclude that the pathogen was introduced to North America at least twice independently. For a new taxonomic concept, we think the complex has to be split into at least two species. New morphological features and further features other than sequence data, however, must be checked for taxonomic value first and, if necessary, be considered.     

New insights into the taxonomy and phylogeny of social voles inferred from mitochondrial cytochrome b sequences

We sequenced the entire cytochrome b gene in Microtus paradoxus from Turkmenistan and Microtus socialis from Crimea and Kalmykia. Phylogenetic relationships among social voles were reconstructed by the inclusion into analyses of a further 23 published haplotypes belonging to six species. The two probabilistic methods which were used in phylogenetic analyses, the Bayesian inference and Maximum Likelihood, yielded very similar results. Both trees showed two highly divergent lineages which were further subdivided into seven species. The socialis lineage encompassed four species (M. socialis, M. irani, M. anatolicus, and M. paradoxus), and the remaining three species clustered into the guentheri lineage (M. guentheri, M. hartingi, M. dogramacii). The ranges for nucleotide divergences between seven species of social voles (4.95–9.28% and 4.18–8.81% for mean and net divergences, respectively) mainly exceeded 4.3%, which is frequently regarded as the conservative cut-off between sibling species in the specious genus Microtus.     

Phylogeography of Japanese population of Phelotrupes auratus (Coleoptera, Geotrupidae) inferred from mitochondrial DNA sequences

A partial sequence of the mitochondrial cytochrome c oxidase subunit I (COI) gene (745 bp) was determined for 57 specimens of a geotrupid beetle (Phelotrupes auratus) from throughout the Japanese archipelago. Of the 57 beetles examined, 42 haplotypes were identified. Phylogenetic trees inferred using maximum parsimony, neighbor joining, and Bayesian inference methods were highly congruent. Reconstructed phylogenetic relationships indicated that P. auratus from the Japanese archipelago was separated into two distinct lineages: Group A, which consisted of 35 haplotypes from Honshu, Shikoku, and Hokkaido Islands, and Group B, which consisted of seven haplotypes from Kyushu and Yakushima Islands. In addition, two sublineages were also recognized within Group A: Subgroup A-1, which consisted of 11 haplotypes from eastern Honshu and Hokkaido, and Subgroup A-2, which consisted of 10 haplotypes from western Honshu and Shikoku. Average genetic distances within Group A were positively correlated to geographic distance between sampling localities. Phylogenetic relationships among haplotypes did not correspond to subspecies classification.     

Ecogeographic correlates of morphometric variation in the Red-billed Chough Pyrrhocorax pyrrhocorax and the Alpine Chough Pyrrhocorax graculus

We estimated the magnitude of intraspecific geographic variation in the Red-billed Chough Pyrrhocorax pyrrhocorax and the Alpine Chough Pyrrhocorax graculus from the analysis of external measurements of museum specimens collected throughout the Palearctic ranges of the species. By means of univariate and multivariate techniques we tested the importance of climatic and geographic factors as potential agents that might have shaped Red-billed and Alpine Chough morphology, discussing the dynamics of the interplay between natural selection, gene flow and phylogenetic constraints. Both species exhibited concordance in character differentiation. Bergmann's rule offered a valid explanation of Red-billed and Alpine Chough body size variation with the largest birds being found at higher elevation, or in colder and more arid regions. Shape was also important. The extremities of the body (bill and tarsus) were longer in warmer areas, in line with Allen's rule. In the Alpine Chough, temperature seemed to be the most important cause of body trait variation, whilst in the Red-billed Chough geographic distance among populations (a measure of geographic isolation) and altitude were also significant. In this paper we critically evaluate the validity of subspecific categories quoted in the literature, given that most variation appeared to be clinal.