Estimating gene flow across hybrid zones: how reliable are microsatellites?

Due to their high polymorphism, microsatellites have become one of the most valued genetic markers in population biology. We review the first two published studies on hybrid zones of the common shrew based on microsatellites. Both reveal surprisingly high interracial gene flow. It can be shown that these are overestimates. Indeed, in classical population genetics models as F-statistics, mutation is neglected. This constitutes an acceptable assumption as long as migration is higher than mutation. However, in hybrid zones where genetic exchanges can be rare, neglecting mutation will lead to strong overestimates of migration when working with microsatellites which display mutation rates up to 10(-3). As there seems to be no straightforward way to correct for this bias, interracial gene flow estimates based on microsatellites should be taken with caution. This problem should however not conceal the enormous potential of microsatellites to unravel the genetics of hybrid zones.

Multilocus phylogeography of the common lizard Zootoca vivipara at the Ibero-Pyrenean suture zone reveals lowland barriers and high-elevation introgression.

BACKGROUND: The geographic distribution of evolutionary lineages and the patterns of gene flow upon secondary contact provide insight into the process of divergence and speciation. We explore the evolutionary history of the common lizard Zootoca vivipara (= Lacerta vivipara) in the Iberian Peninsula and test the role of the Pyrenees and the Cantabrian Mountains in restricting gene flow and driving lineage isolation and divergence. We also assess patterns of introgression among lineages upon secondary contact, and test for the role of high-elevation trans-mountain colonisations in explaining spatial patterns of genetic diversity. We use mtDNA sequence data and genome-wide AFLP loci to reconstruct phylogenetic relationships among lineages, and measure genetic structure RESULTS: The main genetic split in mtDNA corresponds generally to the French and Spanish sides of the Pyrenees as previously reported, in contrast to genome-wide AFLP data, which show a major division between NW Spain and the rest. Both types of markers support the existence of four distinct and geographically congruent genetic groups, which are consistent with major topographic barriers. Both datasets reveal the presence of three independent contact zones between lineages in the Pyrenean region, one in the Basque lowlands, one in the low-elevation mountains of the western Pyrenees, and one in the French side of the central Pyrenees. The latter shows genetic evidence of a recent, high-altitude trans-Pyrenean incursion from Spain into France. CONCLUSIONS: The distribution and age of major lineages is consistent with a Pleistocene origin and a role for both the Pyrenees and the Cantabrian Mountains in driving isolation and differentiation of Z. vivipara lineages at large geographic scales. However, mountain ranges are not always effective barriers to dispersal, and have not prevented a recent high-elevation trans-Pyrenean incursion that has led to asymmetrical introgression among divergent lineages. Cytonuclear discordance in patterns of genetic structure and introgression at contact zones suggests selection may be involved at various scales. Suture zones are important areas for the study of lineage formation and speciation, and our results show that biogeographic barriers can yield markedly different phylogeographic patterns in different vertebrate and invertebrate taxa.

Avifauna do estado do Acre: composição, distribuição geográfica e conservação

O estado do Acre faz fronteiras internacionais com o Peru e a Bolívia e nacionais com os estados do Amazonas e Rondônia. O Acre está localizado nas terras baixas da Amazônia sul‐ocidental, próximo ao sopé dos Andes, dentro de uma região considerada megadiversa da Amazônia brasileira. Apesar disso, a região ainda é pouco conhecida e considerada prioritária para a realização de novos levantamentos biológicos. Com o intuito de contribuir para o conhecimento da avifauna do sudoeste amazônico, este estudo teve como principais objetivos responder as seguintes questões: (a) Quantas e quais são as espécies de aves do estado do Acre? (b) Como as espécies estão distribuídas dentro do estado do Acre? e (c) Qual o estado de conservação das espécies residentes no estado do Acre? A metodologia para responder a estas questões contemplou: (a) uma ampla revisão bibliográfica; (b) dois anos de levantamento em campo, incluindo registros e a coletas de espécimes testemunhos; (c) a confecção do mapa de distribuição de cada táxon (incluindo espécies e subespécies); (d) a distribuição dos táxons pelas três grandes regiões interfluviais do Estado (leste, central e oeste); (e) a identificação de zonas de contato e hibridização, baseada na distribuição dos táxons parapátridos dentro do Estado; (f) o cálculo da distribuição potencial dos táxons dentro do Acre, baseado na extrapolação da área ocupada por cada unidade ecológica (fitofisionomia) onde eles foram registrados; (g) o cálculo da meta de conservação de cada táxon residente no Estado e (h) uma análise de lacunas, baseada na sobreposição dos mapas de distribuição potencial de cada táxon com o das Áreas Protegidas do Estado. A análise de lacuna foi realizada tendo três diferentes cenários como referência: (a) primeiro cenário ‐ levou em consideração todas as Áreas Protegidas; (b) o segundo cenário – levou em consideração apenas as Unidades de Conservação ‐ UCs de Proteção Integral e (c) terceiro cenário – levou em consideração apenas as UCs de Proteção Integral + as de Uso Sustentável (exclusas as Terras Indígenas). A revisão bibliográfica e os levantamentos (históricos e de campo) tiveram início em agosto de 2005 e se estenderam até dezembro de 2007. Após o término da revisão bibliográfica e das expedições em campo, foram compilados 7.141 registros de aves para o todo o estado do Acre. Destes, 4. 623 são de espécimes coletados, dos quais, 2.295 (49,6%) são oriundos de coletas feitas durante a realização deste estudo. Confirmou‐se para o Acre a presença de 655 espécies biológicas, distribuídas em 73 Famílias e 23 Ordens. Como consequência direta deste estudo, cinco novas espécies foram acrescentadas à lista de aves brasileiras. Registrou‐se também, 59 espécies migratórias, das quais, 30 (50,8%) são migrantes neárticas, 11 (18,6%) foram consideradas como migrantes intratropicais e 18 (30,5%) como migrantes austrais. De todas as espécies registradas no Estado, 44 são endêmicas do centro de endemismo Inambari. Dos 556 táxons de aves florestais residentes no Acre, 72,8% (405) distribui‐se nas três sub‐regiões do Estado; 10,0% (56) foi registrado apenas na sub‐região oeste; 5,3% (30) apenas na sub‐região leste e 0,5% (03) apenas na sub‐região central. Ao menos seis pares de táxons irmãos apresentaram padrão de distribuição alopátrida e 15 conjuntos de táxons apresentaram distribuição parapátrida dentro do Estado. Foram identificadas duas zonas de contato secundário (leste/oeste) e duas possíveis zonas de hibridização (leste/oeste) dentro do Estado. As análises de lacunas mostraram que no primeiro cenário, 87,1% dos táxons atingiram 100% da meta de conservação; 12% ficaram em classes de conservação intermediárias, ou seja, em lacuna parcial de proteção e apenas 0,8% ficaram em lacuna total de proteção. No segundo cenário, apenas 0,6% dos táxons atingiram 100% da meta de conservação; 97,6% ficaram em lacuna parcial e 1,8% ficaram em lacuna total de proteção. No terceiro cenário, 73,5% dos táxons atingiram 100% da meta de conservação; 25,5% ficaram em lacuna parcial e apenas 0,8% ficaram em lacuna total de proteção. As principais conclusões obtidas a partir deste estudo foram: (a) que a riqueza avifaunística do estado do Acre é bastante expressiva, porém, o número de espécies detectadas deverá aumentar à medida que novos levantamentos forem realizados; (b) que os rios Purus e Juruá não são as barreiras físicas que determinam o padrão de distribuição da maioria das aves residentes no estado do Acre; (c) que a presença de zonas de contato secundário, não coincidentes com o curso dos dois principais rios do Estado, dá suporte a ideia de que fatores não ligados a uma barreira física devem estar atuando na manutenção do padrão de distribuição atual de alguns táxons de aves residentes no Acre; (d) que o número de espécies “desprotegidas” ou em “lacuna parcial de proteção” entre a avifauna do Acre é muito baixo quando todo o sistema de Áreas Protegidas é levado em consideração, porém este número aumenta com a exclusão das Unidades de Conservação de Uso Sustentável e das Terras Indígenas; (e) que as aves restritas às campinas e campinaranas do oeste do Acre são as únicas que se encontram em lacuna do sistema de áreas protegidas do Acre, indicando a necessidade de se criar uma ou mais Unidades de Conservação para proteger este habitat específico.

Variação geográfica, zonas de intergradação e especiação no complexo Icterus Cayanensis-Chrysocephalus (Aves : Icteridae)

O complexo Icterus cayanensis-chtysocephalus apresenta um intrincado padrão de variação em plumagem e tamanho corpóreo. São reconhecidos, tradicionalmente, para o grupo seis táxons: Icterus chtysocephalus, I. cayanensis cayanensís. I. cayanensis tibialis, I. cayanensis tibialis, I. cayanensis valenciobuenoi, I. cayanensis periporphyrus e I. cayanensis pyrrhopterus, que se substituem geograficamente ao longo de grande parte da América do Sul. Neste estudo foi feita a descrição dos padrões de variação geográfica. Foram diagnosticadas quatro espécies, à luz do conceito filogenético de espécie: Icterus cayanensis (Amazônia Meridional), Icterus chrysocephalus (Amazônia Setentrional), Icterus tibialis (Caatinga) e Icterus pyrrhopterus (Chaco); os táxons I. cayanensis valenciobuenoi e I. cayanensis periporphyrus foram sinonimizados. Entre as formas amazonicas (chrysocephalus e cayanensis) foi detectada a presença de uma zona híbrida mais extensa do que aquela reportada na literatura. No Brasil Central foi diagnosticada a maior zona de intergradação conhecida para aves, com aproximadamente 2.300 km de extensão, produto do intercruzamento entre Icterus tibialis e Icterus pyrrhopterus, formas distribuídas pela Caatinga e Chaco, respectivamente. Postula-se que as zonas de intergradação diagnosticadas neste estudo são produto do intercruzamento de populações previamente diferenciadas em isolamento geográfico.

Tracing reinforcement through asymmetrical partner preference in the European common vole Microtus arvalis

Background The mechanistic basis of speciation and in particular the contribution of behaviour to the completion of the speciation process is often contentious. Contact zones between related taxa provide a situation where selection against hybridization might reinforce separation by behavioural mechanisms, which could ultimately fully isolate the taxa. One of the most abundant European mammals, the common vole Microtus arvalis, forms multiple natural hybrid zones where rapidly diverging evolutionary lineages meet in secondary contact. Very narrow zones of hybridization spanning only a few kilometres and sex-specific gene flow patterns indicate reduced fitness of natural hybrids and incipient speciation between some of the evolutionary lineages. In this study, we examined the contribution of behavioural mechanisms to the speciation process in these rodents by fine-mapping allopatric and parapatric populations in the hybrid zone between the Western and Central lineages and experimental testing of the partner preferences of wild, pure-bred and hybrid female common voles. Results Genetic analysis based on microsatellite markers revealed the presence of multiple parapatric and largely non-admixed populations at distances of about 10 km at the edge of the area of natural hybridization between the Western and Central lineages. Wild females from Western parapatric populations and lab-born F1 hybrids preferred males from the Western lineage whereas wild females of Central parapatric origin showed no measurable preference. Furthermore, wild and lab-born females from allopatric populations of the Western or Central lineages showed no detectable preference for males from either lineage. Conclusions The detected partner preferences are consistent with asymmetrical reinforcement of pre-mating reproductive isolation mechanisms in the European common vole and with earlier results suggesting that hybridization is more detrimental to the Western lineage. As a consequence, these differences in behaviour might contribute to a further geographical stabilization of this moving hybrid zone. Such behavioural processes could also provide a mechanistic perspective for frequently-detected asymmetrical introgression patterns in the largely allopatrically diversifying Microtus genus and other rapidly speciating rodents.

Incipient species formation in salamanders of the Ensatina complex

The Ensatina eschscholtzii complex of plethodontid salamanders, a well-known “ring species,” is thought to illustrate stages in the speciation process. Early research, based on morphology and coloration, has been extended by the incorporation of studies of protein variation and mitochondrial DNA sequences. The new data show that the complex includes a number of geographically and genetically distinct components that are at or near the species level. The complex is old and apparently has undergone instances of range contraction, isolation, differentiation, and then expansion and secondary contact. While the hypothesis that speciation is retarded by gene flow around the ring is not supported by molecular data, the general biogeographical hypothesis is supported. There is evidence of a north to south range expansion along two axes, with secondary contact and completion of the ring in southern California. Current research targets regions once thought to show primary intergradation, but which molecular markers reveal to be zones of secondary contact. Here emphasis is on the subspecies E. e. xanthoptica, which is involved in four distinct secondary contacts in central California. There is evidence of renewed genetic interactions upon recontact, with greater genetic differentiation within xanthoptica than between it and some of the interacting populations. The complex presents a full array of intermediate conditions between well-marked species and geographically variable populations. Geographically differentiated segments represent a diversity of depths of time of isolation and admixture, reflecting the complicated geomorphological history of California. Ensatina illustrates the continuing difficulty in making taxonomic assignments in complexes studied during species formation.

Are there two species of Pygmy Shrews (Sorex)? Revisiting the question using DNA sequence data

Pygmy Shrews in North America have variously been considered to be one species (Sorex hoyi) or two species (S. hoyi and S. thompsoni). Currently, only S. hoyi is recognized. In this study, we examine mitochondrial DNA sequence data for the cytochrome b gene to evaluate the level of differentiation and phylogeographic relationships among eleven samples of Pygmy Shrews from across Canada. Pygmy Shrews from eastern Canada (i.e., Ontario, Quebec, New Brunswick, Nova Scotia, and Prince Edward Island) are distinct from Pygmy Shrews from western Canada (Alberta, Yukon) and Alaska. The average level of sequence divergence between these clades (3.3%) falls within the range of values for other recognized pairs of sister species of shrews. A molecular clock based on third position transversion substitutions suggests that these two lineages diverged between 0.44 and 1.67 million years ago. These molecular phylogenetic data. combined with a reinterpretation of previously published morphological data, are suggestive of separate species status for S. hoyi and S. thompsoni as has been previously argued by others. Further analysis of specimens from geographically intermediate areas (e.g., Manitoba. northern Ontario) is required to determine if there is secondary contact and/or introgression between these two putative species.

Phenotypic divergence but not genetic distance predicts assortative mating among species of a cichlid fish radiation.

The hypothesis of ecological divergence giving rise to premating isolation in the face of gene flow is controversial. However, this may be an important mechanism to explain the rapid multiplication of species during adaptive radiation following the colonization of a new environment when geographical barriers to gene flow are largely absent but underutilized niche space is abundant. Using cichlid fish, we tested the prediction of ecological speciation that the strength of premating isolation among species is predicted by phenotypic rather than genetic distance. We conducted mate choice experiments between three closely related, sympatric species of a recent radiation in Lake Mweru (Zambia/DRC) that differ in habitat use and phenotype, and a distantly related population from Lake Bangweulu that resembles one of the species in Lake Mweru. We found significant assortative mating among all closely related, sympatric species that differed phenotypically, but none between the distantly related allopatric populations of more similar phenotype. Phenotypic distance between species was a good predictor of the strength of premating isolation, suggesting that assortative mating can evolve rapidly in association with ecological divergence during adaptive radiation. Our data also reveals that distantly related allopatric populations that have not diverged phenotypically, may hybridize when coming into secondary contact, e.g. upon river capture because of diversion of drainage systems.

Distinct population structure for co-occurring Anopheles goeldii and Anopheles triannulatus in Amazonian Brazil

To evaluate whether environmental heterogeneity contributes to the genetic heterogeneity in Anopheles triannulatus, larval habitat characteristics across the Brazilian states of Roraima and Pará and genetic sequences were examined. A comparison with Anopheles goeldii was utilised to determine whether high genetic diversity was unique to An. triannulatus. Student t test and analysis of variance found no differences in habitat characteristics between the species. Analysis of population structure of An. triannulatus and An. goeldii revealed distinct demographic histories in a largely overlapping geographic range. Cytochrome oxidase I sequence parsimony networks found geographic clustering for both species; however nuclear marker networks depicted An. triannulatus with a more complex history of fragmentation, secondary contact and recent divergence. Evidence of Pleistocene expansions suggests both species are more likely to be genetically structured by geographic and ecological barriers than demography. We hypothesise that niche partitioning is a driving force for diversity, particularly in An. triannulatus.

Turnover and accumulation of genetic diversity across large time-scale cycles of isolation and connection of populations.

Major climatic and geological events but also population history (secondary contacts) have generated cycles of population isolation and connection of long and short periods. Recent empirical and theoretical studies suggest that fast evolutionary processes might be triggered by such events, as commonly illustrated in ecology by the adaptive radiation of cichlid fishes (isolation and reconnection of lakes and watersheds) and in epidemiology by the fast adaptation of the influenza virus (isolation and reconnection in hosts). We test whether cyclic population isolation and connection provide the raw material (standing genetic variation) for species evolution and diversification. Our analytical results demonstrate that population isolation and connection can provide, to populations, a high excess of genetic diversity compared with what is expected at equilibrium. This excess is either cyclic (high allele turnover) or cumulates with time depending on the duration of the isolation and the connection periods and the mutation rate. We show that diversification rates of animal clades are associated with specific periods of climatic cycles in the Quaternary. We finally discuss the importance of our results for macroevolutionary patterns and for the inference of population history from genomic data.

Forecasting changes in population genetic structure of alpine plants in response to global warming.

Species range shifts in response to climate and land use change are commonly forecasted with species distribution models based on species occurrence or abundance data. Although appealing, these models ignore the genetic structure of species, and the fact that different populations might respond in different ways because of adaptation to their environment. Here, we introduced ancestry distribution models, that is, statistical models of the spatial distribution of ancestry proportions, for forecasting intra-specific changes based on genetic admixture instead of species occurrence data. Using multi-locus genotypes and extensive geographic coverage of distribution data across the European Alps, we applied this approach to 20 alpine plant species considering a global increase in temperature from 0.25 to 4 °C. We forecasted the magnitudes of displacement of contact zones between plant populations potentially adapted to warmer environments and other populations. While a global trend of movement in a north-east direction was predicted, the magnitude of displacement was species-specific. For a temperature increase of 2 °C, contact zones were predicted to move by 92 km on average (minimum of 5 km, maximum of 212 km) and by 188 km for an increase of 4 °C (minimum of 11 km, maximum of 393 km). Intra-specific turnover-measuring the extent of change in global population genetic structure-was generally found to be moderate for 2 °C of temperature warming. For 4 °C of warming, however, the models indicated substantial intra-specific turnover for ten species. These results illustrate that, in spite of unavoidable simplifications, ancestry distribution models open new perspectives to forecast population genetic changes within species and complement more traditional distribution-based approaches.

Fine-scale genetic structure and marginal processes in an expanding population of Biscutella laevigata L. (Brassicaceae).

Evolutionary processes acting at the expanding margins of a species' range are still poorly understood. Genetic drift is considered prevalent in marginal populations, and the maintenance of genetic diversity during recolonization might seem puzzling. To investigate such processes, a fine-scale investigation of 219 individuals was performed within a population of Biscutella laevigata (Brassicaceae), located at the leading edge of its range. The survey used amplified fragment length polymorphisms (AFLPs). As commonly reported across the whole species distribution range, individual density and genetic diversity decreased along the local axis of recolonization of this expanding population, highlighting the enduring effect of the historical colonization on present-day diversity. The self-incompatibility system of the plant may have prevented local inbreeding in newly found patches and sustained genetic diversity by ensuring gene flow from established populations. Within the more continuously populated region, spatial analysis of genetic structure revealed restricted gene flow among individuals. The distribution of genotypes formed a mosaic of relatively homogenous patches within the continuous population. This pattern could be explained by a history of expansion by long-distance dispersal followed by fine-scale diffusion (that is, a stratified dispersal combination). The secondary contact among expanding patches apparently led to admixture among differentiated genotypes where they met (that is, a reshuffling effect). This type of dynamics could explain the maintenance of genetic diversity during recolonization.

Can we continue to neglect genomic variation in introgression rates when inferring the history of speciation? A case study in a Mytilus hybrid zone.

The use of molecular data to reconstruct the history of divergence and gene flow between populations of closely related taxa represents a challenging problem. It has been proposed that the long-standing debate about the geography of speciation can be resolved by comparing the likelihoods of a model of isolation with migration and a model of secondary contact. However, data are commonly only fit to a model of isolation with migration and rarely tested against the secondary contact alternative. Furthermore, most demographic inference methods have neglected variation in introgression rates and assume that the gene flow parameter (Nm) is similar among loci. Here, we show that neglecting this source of variation can give misleading results. We analysed DNA sequences sampled from populations of the marine mussels, Mytilus edulis and M. galloprovincialis, across a well-studied mosaic hybrid zone in Europe and evaluated various scenarios of speciation, with or without variation in introgression rates, using an Approximate Bayesian Computation (ABC) approach. Models with heterogeneous gene flow across loci always outperformed models assuming equal migration rates irrespective of the history of gene flow being considered. By incorporating this heterogeneity, the best-supported scenario was a long period of allopatric isolation during the first three-quarters of the time since divergence followed by secondary contact and introgression during the last quarter. By contrast, constraining migration to be homogeneous failed to discriminate among any of the different models of gene flow tested. Our simulations thus provide statistical support for the secondary contact scenario in the European Mytilus hybrid zone that the standard coalescent approach failed to confirm. Our results demonstrate that genomic variation in introgression rates can have profound impacts on the biological conclusions drawn from inference methods and needs to be incorporated in future studies.

Thirteen polymorphic microsatellite markers for the European green toad Bufo viridis viridis, a declining amphibian species.

We report 13 new polymorphic microsatellite markers for the European green toad Bufo viridis viridis (B. viridis subgroup), a declining amphibian from Central, Southeastern and Eastern Europe. Diversity at these loci estimated for 19 individuals ranged from two to ten alleles. Most of these primers also cross-amplify in related West-Mediterranean green toad species (Bufo balearicus, B. siculus and B. boulengeri). These microsatellites will be useful for conservation genetics of threatened Bufo viridis viridis populations and evolutionary studies of green toad taxa in secondary contact to examine hybridization.

Natural selection in a postglacial range expansion: the case of the colour cline in the European barn owl.

Gradients of variation-or clines-have always intrigued biologists. Classically, they have been interpreted as the outcomes of antagonistic interactions between selection and gene flow. Alternatively, clines may also establish neutrally with isolation by distance (IBD) or secondary contact between previously isolated populations. The relative importance of natural selection and these two neutral processes in the establishment of clinal variation can be tested by comparing genetic differentiation at neutral genetic markers and at the studied trait. A third neutral process, surfing of a newly arisen mutation during the colonization of a new habitat, is more difficult to test. Here, we designed a spatially explicit approximate Bayesian computation (ABC) simulation framework to evaluate whether the strong cline in the genetically based reddish coloration observed in the European barn owl (Tyto alba) arose as a by-product of a range expansion or whether selection has to be invoked to explain this colour cline, for which we have previously ruled out the actions of IBD or secondary contact. Using ABC simulations and genetic data on 390 individuals from 20 locations genotyped at 22 microsatellites loci, we first determined how barn owls colonized Europe after the last glaciation. Using these results in new simulations on the evolution of the colour phenotype, and assuming various genetic architectures for the colour trait, we demonstrate that the observed colour cline cannot be due to the surfing of a neutral mutation. Taking advantage of spatially explicit ABC, which proved to be a powerful method to disentangle the respective roles of selection and drift in range expansions, we conclude that the formation of the colour cline observed in the barn owl must be due to natural selection.