THE EXTENT OF INTROGRESSION OUTSIDE THE CONTACT ZONE BETWEEN NOTROPIS CORNUTUS AND NOTROPIS CHRYSOCEPHALUS (TELEOSTEI: CYPRINIDAE)

The cyprinid fishes, Notropis cornutus and N. chrysocephalus, hybridize in a long, narrow zone in the midwestern United States. To quantify the extent of introgression of genetic markers outside of this zone, samples were collected along transects starting near the region of contact (as defined by morphological characters), followed by samples progressively more distant. Diagnostic allozymic and mitochondrial DNA (mtDNA) restriction site markers were used to estimate the extent of introgression outside of the zone, while polymorphic allozyme and mtDNA markers were used to evaluate the potential for gene flow among populations within transects. Analysis of populations from the northern transect provided evidence for differentiation of populations for some of the markers; however, on average, enough gene flow has occurred to overcome substantial differentiation. Introgressed mtDNA and allozyme haplotypes were rare and found only in the population closest to the contact zone. The rarity of introgressed alleles in the more northern populations is consistent with the recent origin of these populations after the Wisconsin glaciation (less than 12,000 years bp) and/or selection maintaining the northern boundary of the contact zone. Analysis of populations from the southern transect revealed evidence for population subdivision but no evidence for introgression at the diagnostic allozyme loci; however, nearly all individuals from this transect possessed introgressed mtDNA haplotypes, with samples furthest from the contact zone exhibiting the highest frequencies of introgression. Patterns of variation for one of the polymorphic allozyme markers (Est-A) and introgressed mtDNAs were highly correlated, suggesting that allozymic heterogeneity at this locus is also the result of introgression. The most likely explanation for these data is that these introgressed haplotypes are indicators of a more southern position of the contact zone during the Pleistocene, with the contact zone shifting northward with the recession of the glacial front. Such movement implicates selection in the maintenance of distributional limits of these species, and hence, the width and position of the contact zone.     

Mitochondrial DNA diversity in an apomictic Daphnia complex from the Canadian High Arctic

Cyclic parthenogenesis is the ancestral mode of reproduction in the cladoceran crustacean, Daphnia pulex, but some populations have made the transition to obligate parthenogenesis and this is the only mode of reproduction known to occur in arctic populations. Melanism and polyploidy are also common in arctic populations of this species. Prior allozyme studies of arctic D. pulex revealed substantial levels of clonal diversity on a regional scale. Clonal groupings based on cluster analysis of allozyme genotypes do not conform to groupings based on the presence/absence of melanin or on ploidy level. In order to further elucidate genetic relationships among arctic D. pulex clones, mitochondrial DNA (mtDNA) variation was examined in 31 populations from two Canadian high-arctic sites. The data were also compared to a previous study of mtDNA variation in populations from a Canadian low-arctic site. Cladistic analysis of restriction site variation of the entire mitochondrial genome and nucleotide sequence variation of the mitochondrial control region was used to construct genetic relationships among mitochondrial genotypes. Three distinct mitochondrial lineages were detected. One lineage was associated with diploid, nonmelanic clones and is the same as the lineage that is found in temperate populations of D. pulex. The other two lineages (A & B) were associated with polyploid, melanic clones. Sequence divergence between the A and B lineages was 2.4%. Sequence divergence between D. pulex and either of these two lineages exceeded 3%. It is suggested that the melanic, polyploid clones are hybrids between males of D. pulex (and/or a closely related congener, D. pulicaria) and females of either of two ancestral melanic species that have mitochondrial lineages A and B. Geographic patterns of mitochondrial diversity in ‘melanic’ lineage B support the hypothesis of an high-arctic refuge for the ancestral species during the last glacial period.     

Origins of clonal diversity in the hypervariable asexual ostracode Cypridopsis vidua

Prior allozyme studies have indicated that populations of the asexual ostracode, Cypridopsis vidua (Müller), show extraordinary clonal diversity. Based on a joint examination of allozyme variation and sequence divergence at the mitochondrial cytochrome c oxidase I (COI) gene, the present analysis provides new insights concerning the origins of this variation. The results establish that populations of C. vidua in one recently deglaciated region of North America are not only allozymically diverse, but also include several divergent mitochondrial DNA (mtDNA) lineages. The extent of sequence divergence among these lineages is so large as to suggest their diversification over the past 7–8 million years. The patterning of genetic divergence among co-occurring clones makes it apparent that much of the mtDNA and allozyme diversity in local populations owes its origins to recurrent colonization events. However, in situ mutational diversification also appears to explain some variation. The mechanisms enabling the sustained coexistence of such a large array of closely allied genotypes remain unclear, but there is an apparent difference in equilibrium diversity between benthic and planktonic asexual organisms.     

Differentiation in morphology and habitat partitioning of genetically characterized Gammarus fossarum forms (Amphipoda) across a contact zone

Evolutionary processes do not affect all character types equally. Depending on environmental effects, morphological and ecological evolution may be uncoupled from molecular evolution. A natural contact zone between two forms of the amphipod Gammarus fossarum was used to assess the levels of differentiation in genetic, morphological and habitat characters. About 50 individuals per population were analysed for six enzyme loci in 72 populations and four size-standardized morphometric traits in a subset of 32 populations. Nine environmental parameters per population were used to characterize the habitat. All three character sets revealed significant divergences between G. fossarum forms. The eastern form A of G. fossarum differs from the western form B by longer setae at the second antennae (males) and longer exopodits at die third uropods (males and females). The variance partitioning within populations, among populations and between forms, was compared within the three character sets. The relative variance component between forms in genetic characters appeared to be 10-fold greater in comparison to morphological traits, indicating morphological stasis during speciation. The inter-form environmental differentiation was shown to be low, but consistent between pure populations. The eastern form A seems to prefer higher altitudes, a substrate type of gravel, stones and leaves, shading, and water with higher conductivity and pH man the western form B.     

EVIDENCE FOR A MOSAIC HYBRID ZONE IN THE GRASS SHRIMP PALAEMONETES KADIAKENSIS (PALAEMONIDAE) AS REVEALED BY MULTIPLE GENETIC MARKERS

Molecular techniques provide powerful tools for studying the geographic structure of hybrid zones and the dynamics of gene exchange between incipient species. We examined allozyme variation at five loci (PGM, GPI, MDH-1, MDH-2, and LDH) for 27 populations of Palaemonetes kadiakensis from the central, coastal, and eastern regions of Texas. Central Texas populations of P. kadiakensis exhibited highly significant linkage disequilibrium and departures from Hardy-Weinberg genotype proportions. In populations with linkage disequilibrium, allelic differences at GPI defined two types of P. kadiakensis, designated A and B. Both types existed in central Texas with little or no evidence of interbreeding, whereas the populations from all other localities showed complete introgression of type B alleles into the type A gene pool. We also examined ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) variation in a subset of populations, chosen to cover a range of geographic locations and levels of linkage disequilibrium. Two groups of mtDNA haplotypes and two restriction fragment patterns for the rDNA corresponded to allozyme type A and B individuals in populations exhibiting linkage disequilibrium. In populations with ongoing hybridization, all hybrid animals (N= 15) exhibited type A mtDNA. Exhibition of type A mtDNA indicated that type A females had mated successfully with type B males, but type B females had not mated successfully with type A males. Genotype distributions suggest reduced reproduction by hybrid offspring in central Texas populations. These patterns are consistent with a mosaic model of hybrid zone dynamics.     

Individual performance in relation to cytonuclear discordance in a northern contact zone between long‐toed salamander (Ambystoma macrodactylum) lineages

Cytonuclear discordance in contact zones between related lineages is common, with mitochondrial clines often being displaced from clines in nuclear allele frequency. Proposed explanations for such a pattern include adaptive introgression of mtDNA or a neutral wake of mtDNA being left behind following hybrid zone movement. However, studies investigating these hypotheses are rare. Our previous survey of genetic variation in the long‐toed salamander (Ambystoma macrodactylum) highlighted a potential case of cytonuclear discordance between two lineages in western Canada. Here, we use additional markers and samples to clarify the extent of this discordance. We simultaneously assess the feeding performance of individuals in a common environment to test for an association between mitotype and individual performance. The genetic results confirm a general pattern of cytonuclear discordance in the focal region. However, we also observed more limited introgression of a diagnostic nuclear marker. Intriguingly, although there were differences in individual performance associated with the transition between mitotypes, these differences were not fully explained by mitotype. Instead, the lowest performance was observed in individuals demonstrating the greatest mismatch between mtDNA and all nuclear markers, suggesting the potential for cytonuclear incompatibilities to be acting. These results highlight the complexity of understanding the causes and consequences of mtDNA introgression and cytonuclear discordance in contact zones.     

Spatial distribution of cryptic species diversity in european freshwater amphipods (Gammarus fossarum) as revealed by pyrosequencing

In order to understand and protect ecosystems, local gene pools need to be evaluated with respect to their uniqueness. Cryptic species present a challenge in this context because their presence, if unrecognized, may lead to serious misjudgement of the distribution of evolutionarily distinct genetic entities. In this study, we describe the current geographical distribution of cryptic species of the ecologically important stream amphipod Gammarus fossarum (types A, B and C). We use a novel pyrosequencing assay for molecular species identification and survey 62 populations in Switzerland, plus several populations in Germany and eastern France. In addition, we compile data from previous publications (mainly Germany). A clear transition is observed from type A in the east (Danube and Po drainages) to types B and, more rarely, C in the west (Meuse, Rhone, and four smaller French river systems). Within the Rhine drainage, the cryptic species meet in a contact zone which spans the entire G. fossarum distribution range from north to south. This large-scale geographical sorting indicates that types A and B persisted in separate refugia during Pleistocene glaciations. Within the contact zone, the species rarely co-occur at the same site, suggesting that ecological processes may preclude long-term coexistence. The clear phylogeographical signal observed in this study implies that, in many parts of Europe, only one of the cryptic species is present.     

Genetic differentiation and history of populations of the Italian treefrog Hyla intermedia: lack of concordance between mitochondrial and nuclear markers

The genetic differentiation among 33 populations of the Italian treefrog, Hyla intermedia (Anura: Hylidae), was investigated using both biparentally (23 allozyme loci) and maternally (partial mitochondrial cytochrome b gene) inherited markers. Two main population groups were evidenced by both markers, located north and south of the northern Apennines. However, the pattern of differentiation between these two groups was much less pronounced at allozymes than at mtDNA, leading to gene flow estimates that were 25 times lower at mitochondrial than at nuclear level. Also, the mtDNA divergence between the two groups was particularly marked for two cospecific lineages of anuran amphibians (the P-distance being on average 9.04%), while their average genetic distance at allozymes was comparatively low (D _NEI = 0.07). This contrasting pattern of nuclear versus mitochondrial genetic variation is discussed in the context of: (1) marker specific selection, (2) secondary contact and sex-biased gene flow and (3) ancestral polymorphism and colonization from north to south. Finally we emphasize how, for population genetic studies, the use of multiple markers having distinct evolutionary properties can help unravel the existence of more complex evolutionary histories.     

Behavioural Reproductive Isolation in a Rotifer Hybrid Zone

A hybrid zone between two Brachionus plicatilis rotifer mitochondrial DNA (mtDNA) lineages was recently described in the Iberian Peninsula between a pond (Santed 2) and a lake (Gallocanta). The patterns of mitochondrial and nuclear genetic variation observed suggested that gene flow is mainly male-mediated from the lake to the pond. Here we test two hypotheses: (a) that male-mediated gene flow occurs through assortative mating between individuals from these ponds, (b) that behavioural isolation occurs between the two mtDNA lineages. We isolated, reared and genotyped rotifer clones from resting eggs collected in the sediments of these and two other distant ponds. We devised a quick, inexpensive RFLP method to discriminate between B. plicatilis and its sibling species B. ‘Manjavaeas’ and between both mtDNA B. plicatilis lineages. Behavioural no-choice tests using new-born, virgin males and females were performed between five clones. B. ‘Manjavacas’ and B. plicatilis were reproductively isolated. B. plicatilis clones did not show evidence of reproductive isolation, regardless of their mtDNA lineage, except Santed 2 males, which discriminated strongly against Gallocanta females. These results could help to explain the discrepancies between mitochondrial and nuclear genetic variation reported in the two populations.     

Phylogeography of Bufo fowleri at its northern range limit

Many of the species that recolonized previously glaciated areas in the Great Lakes basin of North America over the past 10-12,000 years exhibit genetic evidence of multiple invasion routes and present-day secondary contact between deeply divergent lineages. With this in mind, we investigated the phylogeographical structure of genetic variability in Fowler's toads (Bufo fowleri) at the northern edge of its distribution where its range encircles the Lake Erie basin. Because B. fowleri is so closely tied to habitats along the Lake Erie shoreline, we would expect to find clear evidence of the number of invasions leading to the species' colonization of the northern shore. A 540 bp sequence from the mitochondrial control region was amplified and analysed for 158 individuals from 21 populations. Interpopulation sequence variation ranged from 0% to 6%. Phylogenetic analysis of p-distance using the neighbor-joining method revealed two deeply divergent (6% sequence divergence) mtDNA lineages (Phylogroup 1 and 2), possibly arising as a result of secondary contact of populations that entered the region from two separate glacial refugia. However, the phylogeographical pattern was not simple. The populations at Long Point, on the north shore of Lake Erie, clustered with the population from Indiana Dunes on Lake Michigan to form Phylogroup 2 whereas all other B. fowleri populations examined from both sides of Lake Erie constituted Phylogroup 1. Furthermore, mtDNA sequences from the related species Bufo americanus, obtained from populations outside the range of B. fowleri, clustered with mtDNA haplotypes of B. fowleri Phylogroup 1, indicating the possibility of partial introgression of mitochondria from one species to the other.     

Genealogy of the nuclear beta-fibrinogen locus in a highly structured lizard species: comparison with mtDNA and evidence for intragenic recombination in the hybrid zone

The study of nuclear genealogies in natural populations of nonmodel organisms is expected to provide novel insights into the evolutionary history of populations, especially when developed in the framework of well-established mtDNA phylogeographical scenarios. In the Iberian Peninsula, the endemic Schreiber's green lizard Lacerta schreiberi exhibits two highly divergent and allopatric mtDNA lineages that started to split during the late Pliocene. In this work, we performed a fine-scale analysis of the putative mtDNA contact zone together with a global analysis of the patterns of variation observed at the nuclear beta-fibrinogen intron 7 (beta-fibint7). Using a combination of DNA sequencing with single-strand conformational polymorphism (SSCP) analysis, we show that the observed genealogy at the beta-fibint7 locus reveals extensive admixture between two formerly isolated lizard populations while the two mtDNA lineages remain essentially allopatric. In addition, a private beta-fibint7 haplotype detected in the single population where both mtDNA lineages were found in sympatry is probably the result of intragenic recombination between the two more common and divergent beta-fibint7 haplotypes. Our results suggest that the progressive incorporation of nuclear genealogies in investigating the ancient demography and admixture dynamics of divergent genomes will be necessary to obtain a more comprehensive picture of the evolutionary history of organisms.     

Low, but Strongly Structured Mitochondrial DNA Diversity in Root Knot Nematodes (Meloidogyne)

Root-knot nematodes (genus Meloidogyne) have been the subject of recent and numerous studies of genetic variation because of the need to develop molecular diagnostics for the four globally distributed, parthenogenetic species that are significant agricultural pests. Our analysis of Meloidogyne mtDNA improves on previous studies: (i) by examining restriction site polymorphism among a large number of isolates also characterized for standard morphological, host range and allozyme phenotypes; (ii) by using higher resolution electrophoretic techniques; and (iii) by mapping variable restriction sites with reference to the complete nucleotide sequence. This revealed fivefold less sequence divergence (<0.6%) between variants than estimated in previous restriction fragment length polymorphism (RFLP) studies, but perfect correspondence between mtDNA haplotype and allozyme (esterase) phenotypes. The mtDNA variation, although limited, is strongly structured with as much divergence between two lineages of Meloidogyne arenaria as between either of these and Meloidogyne javanica. The low diversity of mtDNAs suggests that these parthenogenetic lineages arose from distinct but closely related sexual females, a pattern seen in other parthenogenetic complexes. In contrast to the concordance between mtDNA and allozyme markers, there were several discrepancies between the traditional methods of identification. We suggest that further studies of these nematodes should focus on well defined genetic groups, whether or not these coincide with existing taxonomic units.     

A single mitochondrial lineage is shared by morphologically and allozymatically distinct freshwater Corbicula clones

Despite that the exotic invasion and rapid range expansion of Asian freshwater Corbicula into new environments have been of intensive research topic in freshwater ecology, the genetic structures of freshwater Corbicula in its native range remain poorly understood. In this study, the genetic structures of two Korean freshwater Corbicula clonal lineages were characterized by cross-referencing the nuclear genomic structures with mtDNA sequence analysis. In spite of substantial genetic differences (Nei's D = 0.363-0.372) and a pronounced level of fixed allelic distinctions (in six of 20 allozyme loci) between Corbicula lineages, no lineage-specific mtDNA differentiation was observed. The evident disjunction between mtDNA sequences and nuclear genomes is a compelling evidence for the existence of interspecific nuclear hybrid genome structures, comprising different combinations of paternal and maternal contributions. This unusual novel finding is the first case demonstrating that morphologically and allozymatically distinct, yet mitochondrially identical clonal lineages exist in the genus Corbicula. However, we could not find the ancestral species for these two clonal lineages in the present study, and the answer for this question must wait until the genetic structure of Asian Corbicula taxa is fully characterized.     

Mitochondrial DNA variability in viviparous and ovoviviparous populations of the urodele Salamandra salamandra

Mitochondrial DNA analysis of 13 populations of S. salamandra along a transect across the North of the Iberian Peninsula showed values of divergence between haplotypes ranging from d = 0.41% to 5.91%. Phenetic and cladistic analysis grouped the isofemale lineages into two main clusters with contrasting phylogeographic patterns. The first group encompasses populations located at each extreme of the Iberian Peninsula. Despite the large geographic distance separating these populations, they exhibit only a minor degree of divergence among haplotypes. In contrast, much higher diversification, in both number of distinct haplotypes, and overall genetic divergence, was observed in the second group of phylogenetically related populations. Surprisingly, this process of radiation and divergence in mtDNA haplotypes occurred in populations in close geographic proximity. All populations sampled in this group are located within a 300 km range, in the central part of our transect across the Northern edge of the Peninsula. Most populations in the central range of our transect exhibit viviparous reproduction — which is derived and highly unusual among urodeles. The genetic distances measured among Asturian (central portion of our transect), viviparous populations are higher than the distances measured between the two main taxonomic clusters. A viviparous population showing an unusual level of mtDNA heterogenetiy is reported and the potential implications of this focus of localized variability are discussed. The dynamics of isofemale lineages among the two reproductive modes was further explored in combination with the previous allozyme data. Several nuclear markers suggest that major mtDNA divergences could be explained by long-term extrinsic barriers to gene flow. Isofemale lineages indicate a narrow secondary contact zone among populations with different reproductive patterns. The existence of viviparous and ovovivparous populations sharing a common haplotype suggests that reproductive transition in S. salamandra could have arisen in absence of genetic mtDNA differentiation. We finally outline a genetic model system where the acquisition of water independence from a primitively aquatic dependent amphibian life cycle can be analyzed from a microevolutionary perspective.     

Biogeography of the túngara frog, Physalaemus pustulosus: a molecular perspective

Physalaemus pustulosus, a small leptodactylid frog with South American affinities, ranges across northern South America through Middle America to southern Mexico. To investigate its geographic variation and evolutionary origins, we analysed the presumptive gene products of 14 allozyme loci and sequenced a portion of the mitochondrial COI gene from individuals sampled throughout the distribution. Generally, allozyme dissimilarities and sequence divergences are correlated with each other and with geographic proximity. The greatest discontinuity in genetic variation was found between populations in Middle America vs. South America + Panama. Based on two Bayesian MCMC (Markov chain Monte Carlo) divergence time estimates involving two independent temporal constraints, the timing of the separation of northern and southern túngara frog lineages is significantly older than the time since completion of the current Panama land bridge. P. pustulosus first invaded Middle America from South America about 6-10 million years ago giving rise to the northern lineage. The southern lineage then invaded Panama independently after land bridge completion. Despite millions of years of independent evolution, the multilocus allozyme data revealed that western Panama populations represent a contact zone containing individuals with alleles from both groups present.     

Contrasting patterns of mitochondrial DNA and microsatellite introgressive hybridization between lineages of lake whitefish (Coregonus clupeaformis); relevance for speciation

We performed a combined analysis of mitochondrial DNA (mtDNA) and microsatellite loci among lake whitefish (Coregonus clupeaformis) populations in order to assess the levels of congruence between both types of markers in defining patterns of genetic structuring, introgressive hybridization and inferring population origins in the hybrid zone of the St. John River basin. A second objective was to test the hypothesis that secondary contact between glacial lineages always resulted in the occurrence of sympatric dwarf and normal whitefish ecotypes. Fish were sampled from 35 populations and polymorphism was screened at mtDNA and six microsatellite loci for a total of 688 and 763 whitefish, respectively. Four lakes harbouring a single whitefish population of normal ecotype admixed with mtDNA haplotypes of different lineages were found. This confirmed that secondary contact between whitefish evolutionary lineages did not always result in the persistence of reproductively isolated ecotypes. Microsatellites further supported the definition of distinct glacial lineages by identifying lineage-specific allelic size groups. They also further supported the hypothesis that ecotypes originated from either a single founding lineage (sympatric divergence) or following secondary contacts between lineages (allopatric divergence), depending on the lake. In general, however, the pattern of population differentiation and introgressive hybridization observed at microsatellites was in sharp contrast with that depicted by mtDNA variation. Both factorial correspondence analysis and analysis of admixture proportion revealed a much more pronounced pattern of introgressive hybridization than depicted by mtDNA analyses. Variable levels of introgression indicated that environmental differences may be as important as the historical contingency of secondary contact in explaining the persistence of sympatric ecotypes and the differential pattern of introgressive hybridization among lakes. Whitefish populations from the St. John River basin hybrid zone represent a rare illustration of a continuum of both morphological and genetic differentiation within a given taxon, spanning from complete introgression to possibly complete reproductive isolation, depending on lakes. Thus, each lake may be viewed as a different temporal snapshot taken throughout the gradual process of speciation.     

Complete mitochondrial DNA analysis of eastern Eurasian haplogroups rarely found in populations of northern Asia and eastern Europe

With the aim of uncovering all of the most basal variation in the northern Asian mitochondrial DNA (mtDNA) haplogroups, we have analyzed mtDNA control region and coding region sequence variation in 98 Altaian Kazakhs from southern Siberia and 149 Barghuts from Inner Mongolia, China. Both populations exhibit the prevalence of eastern Eurasian lineages accounting for 91.9% in Barghuts and 60.2% in Altaian Kazakhs. The strong affinity of Altaian Kazakhs and populations of northern and central Asia has been revealed, reflecting both influences of central Asian inhabitants and essential genetic interaction with the Altai region indigenous populations. Statistical analyses data demonstrate a close positioning of all Mongolic-speaking populations (Mongolians, Buryats, Khamnigans, Kalmyks as well as Barghuts studied here) and Turkic-speaking Sojots, thus suggesting their origin from a common maternal ancestral gene pool. In order to achieve a thorough coverage of DNA lineages revealed in the northern Asian matrilineal gene pool, we have completely sequenced the mtDNA of 55 samples representing haplogroups R11b, B4, B5, F2, M9, M10, M11, M13, N9a and R9c1, which were pinpointed from a massive collection (over 5000 individuals) of northern and eastern Asian, as well as European control region mtDNA sequences. Applying the newly updated mtDNA tree to the previously reported northern Asian and eastern Asian mtDNA data sets has resolved the status of the poorly classified mtDNA types and allowed us to obtain the coalescence age estimates of the nodes of interest using different calibrated rates. Our findings confirm our previous conclusion that northern Asian maternal gene pool consists of predominantly post-LGM components of eastern Asian ancestry, though some genetic lineages may have a pre-LGM/LGM origin.     

Phylogenetic Relationships Among Fragmented Asian Black Bear (Ursus Thibetanus) Populations in Western Japan

The number of Asian black bears (Ursus thibetanus) in Japan has been reduced and their habitats fragmented and isolated because of human activities. Our previous study examining microsatellite DNA loci revealed significant genetic differentiation among four local populations in the western part of Honshu. Here, an approximate 700-bp nucleotide sequence of mitochondrial DNA (mtDNA) control region was analysed in 119 bears to infer the evolutionary history of these populations. Thirteen variable sites and variation in the number of Ts at a T-repeat site were observed among the analysed sequences, which defined 20 mtDNA haplotypes with the average sequence divergence of 0.0051 (SD = 0.00001). The observed haplotype frequencies differed significantly among the four populations. Phylogeographic analysis of the haplotypes suggested that black bears in this region have gone through two different colonisation histories, since the observed haplotypes belonged to two major monophyletic lineages and the lineages were distributed with an apparent border. The spatial genetic structure revealed by using mtDNA was different from that observed using microsatellite DNA markers, probably due to female philopatry and male-biased dispersal. Since nuclear genetic diversity will be lost in the three western populations because of the small population size and genetic isolation, their habitats need to be preserved, and these four populations should be linked to each other by corridors to promote gene flow from the easternmost population with higher nuclear genetic diversity.     

Simple identification of mitochondrial lineages in contact zones based on lineage-selective primers

A variety of research projects focus on genetic variation among and within maternal lineages as encompassed by mitochondrial DNA (mtDNA). While mtDNA often differs substantially between species, large differences may also be found within species. The evaluation of such divergent lineages, for example in intraspecific contact zones (hybrid zones), commonly involves sequencing numerous individuals. Large‐scale sequencing is both expensive and labour‐intensive. Based on sequences from 15 individuals, we devised a simple and quick polymerase chain reaction assay for identification of divergent mtDNA lineages in a secondary contact zone of the side‐blotched lizard (Uta stansburiana). The application uses lineage‐selective primers to amplify a lineage‐diagnostic product, and is based on each group of mtDNA haplotypes being a monophyletic assemblage of haplotypes sharing the same maternal ancestry, deeply divergent from the other group. The assay was tested on a larger sample (n = 147) of specimens from the contact zone, confirming its usefulness in quick and reliable identification of mtDNA lineages. This approach can be modified for other species, provided diagnostic lineage variation is available, and may also be performed in simple laboratory settings while conducting fieldwork.     

Genetic structure and colonization processes in European populations of the common vole, Microtus arvalis

The level of genetic differentiation within and between evolutionary lineages of the common vole (Microtus arvalis) in Europe was examined by analyzing mitochondrial sequences from the control region (mtDNA) and 12 nuclear microsatellite loci (nucDNA) for 338 voles from 18 populations. The distribution of evolutionary lineages and the affinity of populations to lineages were determined with additional sequence data from the mitochondrial cytochrome b gene. Our analyses demonstrated very high levels of differentiation between populations (overall FST: mtDNA 70%; nucDNA 17%). The affinity of populations to evolutionary lineages was strongly reflected in mtDNA but not in nucDNA variation. Patterns of genetic structure for both markers visualized in synthetic genetic maps suggest a postglacial range expansion of the species into the Alps, as well as a potentially more ancient colonization from the northeast to the southwest of Europe. This expansion is supported by estimates for the divergence times between evolutionary lineages and within the western European lineage, which predate the last glacial maximum (LGM). Furthermore, all measures of genetic diversity within populations increased significantly with longitude and showed a trend toward increase with latitude. We conclude that the detected patterns are difficult to explain only by range expansions from separate LGM refugia close to the Mediterranean. This suggests that some M. arvalis populations persisted during the LGM in suitable habitat further north and that the gradients in genetic diversity may represent traces of a more ancient colonization of Europe by the species.