PATTERNS OF VARIATION AND LINKAGE DISEQUILIBRIUM IN A FIELD CRICKET HYBRID ZONE

The distribution of multilocus genotypes found within a natural hybrid zone is determined by the sample of genotypes present when the hybrid zone first formed, by subsequent patterns of genetic exchange between the hybridizing taxa, and by drift and selection within each of the hybrid zone populations. We have used anonymous nuclear DNA restriction fragment polymorphisms (RFLPs) to characterize the array of multilocus genotypes present within a well-studied hybrid zone between two eastern North American field crickets, Gryllus pennsylvanicus and Gryllus firmus. These crickets hybridize along a zone of contact that extends from New England to Virginia. Previous studies have shown that both premating and postmating barriers exist between the two cricket species, but the absence of diagnostic morphological and allozyme markers has made it difficult to assess the consequences of these barriers for genetic exchange. Analyses based on four diagnostic anonymous nuclear markers indicate that hybrid zone populations in Connecticut contain few F₁ hybrids, and that nonrandom associations persist among nuclear gene markers, between nuclear and cytoplasmic markers, and between molecular markers and morphology. Field cricket populations within the hybrid zone are not “hybrid swarms” but consist primarily of crickets that are very much like one or the other of the parental species. Despite ample opportunity for genetic exchange and evidence for introgression at some loci, the two species remain quite distinct. Such a pattern appears to be characteristic of many natural hybrid zones.     

Concordance of genetic and phenotypic characters across a sapsucker hybrid zone

Hybridization has presented a challenge for taxonomists and conservation biologists, since hybridizing forms could be stable evolutionary entities or ephemeral forms that are blending together. However, hybrid zones also provide a unique opportunity for evolutionary biologists who study the interaction between gene flow and reproductive isolation in speciation. Three forms of woodpeckers (sapsuckers; genus Sphyrapicus) in North America that are mostly geographically separated but hybridize with each other where they come into contact present a remarkable system for the study of hybridization. We provide the first comprehensive analysis of phenotypic and genetic variation across a hybrid zone between two of these forms, the red‐breasted Sphyrapicus ruber and yellow‐bellied S. varius sapsuckers. The objective was to infer whether selection maintains the differences between forms. Our analysis of eight morphometric and 20 plumage traits, and two molecular markers showed clear differences between the forms and roughly concordant clinal variation across a narrow hybrid zone. Thirty percent of sampled birds in the hybrid zone had mixed west/east genotypes at the genetic markers examined. The center of the genetic cline was located 20 km west of the crest of the Rocky Mountains. The width of the zone was 122 km, narrower than would be expected under neutral blending given reasonable estimates of the age of the zone and individual dispersal distances. Heterozygote deficit and cytonuclear disequilibrium at the centre of the hybrid zone suggested nonrandom mating or limited hybridization. Given these patterns and lack of evidence for habitat segregation we conclude that this hybrid zone is maintained by selection, most likely in the form of hybrid inferiority. This study provides an illustrative example of extensive hybridization between stable entities, providing additional evidence against the historical practice of treating hybridizing forms as members of the same species.     

Introgressive hybridization and morphological transgression in the contact zone between two Mediterranean Solea species

Hybrid zones provide natural experiments where new combinations of genotypes and phenotypes are produced. Studying the reshuffling of genotypes and remodeling of phenotypes in these zones is of particular interest to document the building of reproductive isolation and the possible emergence of transgressive phenotypes that can be a source of evolutionary novelties. Here, we specifically investigate the morphological variation patterns associated with introgressive hybridization between two species of sole, Solea senegalensis and Solea aegyptiaca. The relationship between genetic composition at nuclear loci and individual body shape variation was studied in four populations sampled across the hybrid zone located in northern Tunisia. A strong correlation between genetic and phenotypic variation was observed among all individuals but not within populations, including the two most admixed ones. Morphological convergence between parental species was observed close to the contact zone. Nevertheless, the samples taken closest to the hybrid zone also displayed deviant segregation of genotypes and phenotypes, as well as transgressive phenotypes. In these samples, deviant body shape variation could be partly attributed to a reduced condition index, and the distorted genetic composition was most likely due to missing allelic combinations. These results were interpreted as an indication of hybrid breakdown, which likely contributes to postmating reproductive isolation between the two species.     

Development of RAPD markers in two introduced fire ants, Solenopsis invicta and S. richteri, and their application to the study of a hybrid zone

We developed RAPD DNA markers useful in distinguishing the fire ants Solenopsis invicta and S. richteri. An initial survey of 200 primers revealed seven informative markers; family studies allowed us to determine expression patterns and to confirm Mendelian inheritance of these markers. The seven RAPD markers, one of which is inherited as a codominant marker, were employed along with three allozyme markers to describe the structure of a hybrid zone that has formed between the two species in the USA, where they have been introduced. The data suggest minor introgression of alleles from one parental species ( S. richteri ) into the other ( S. invicta ), which most likely reflects the documented recent movement of this hybrid zone. This pattern is interpreted as interspecific introgression rather than shared intraspecific polymorphism on the basis of comparisons with samples from native, non-hybridizing populations in South America that lack such polymorphism. The data further reveal that the structure of the hybrid zone in the USA varies geographically. One parental species ( S. invicta ) and the hybrids exhibit a mosaic distribution in the east; a gradual transition between the parental species occurs in the centre, with a large intervening area of hybrid genotypes only; and there is apparent contact between parental populations with a small or no intervening zone of hybridization in the west. These differing patterns in the structure of the hybrid zone presumably reflect the unique histories of colonization in different parts of the range of introduced fire ants.     

PATTERNS OF HYBRIDIZATION IN THE PIRIQUETA CAROLINIANA COMPLEX IN CENTRAL FLORIDA: EVIDENCE FOR AN EXPANDING HYBRID ZONE

We have identified a broad zone of hybridization between two morphologically and ecologically distinct herbaceous perennial taxa (morphotypes) within the Piriqueta caroliniana complex, which extends more than 300 km across the central Florida peninsula. Phylogeographic analyses indicate that the caroliniana morphotype has been present in north and central Florida since the early Pleistocene and that the viridis morphotype has immigrated into southern Florida much more recently. We examine the distribution of diagnostic morphological characters and nuclear genetic markers to assess the extent and patterns of introgression in this system. The results are consistent with the hypothesis that this hybrid zone has expanded north in recent history as viridis alleles have introgressed into regions that were previously occupied by populations of caroliniana. Genetic markers diagnostic for caroliniana have consistently high frequencies across the hybrid zone, whereas markers for viridis are extremely variable among populations with frequency reversals in adjacent populations. The latter pattern is probably the result of the combined stochastic effects of dispersal and drift on viridis alleles as they introgressed northward. Additional evidence for the recent expansion of this hybrid zone comes from patterns of variation for morphological and genetic markers. As expected for an expanding hybrid zone, within-population morphological variation was greatest toward the advancing front of introgression and levels of genetic variation for neutral diagnostic markers were greatest in the region of initial contact and lower in areas of recent expansion. The observed patterns of variation suggest that at least some hybrid genotypes have high fitnesses, which has led to the expansion of the hybrid zone via the displacement of parental genotypes in central Florida.     

Low levels of hybridization across two contact zones among three species of woodpeckers (Sphyrapicus sapsuckers)

Three species of closely related woodpeckers (sapsuckers; Sphyrapicus) hybridize where they come into contact, presenting a rare ‘λ‐shape’ meeting of hybrid zones. Two of the three arms of this hybrid zone are located on either side of the Interior Plateau of British Columbia, Canada bordering the foothills of the Coast Mountains and the Rocky Mountains. The third arm is located in the eastern foothills of the Rocky Mountains. The zones of hybridization present high variability of phenotypes and alleles in relatively small areas and provide an opportunity to examine levels of reproductive isolation between the taxa involved. We examined phenotypes (morphometric traits and plumage) and genotypes of 175 live birds across the two hybrid zones. We used the Genotyping By Sequencing (GBS) method to identify 180 partially diagnostic single nucleotide polymorphisms (SNPs) to generate a genetic hybrid index (GHI) for each bird. Phenotypically diverged S. ruber and S. nuchalis are genetically closely related, while S. nuchalis and S. varius have similar plumage but are well separated at the genetic markers studied. The width of both hybrid zones is narrower than expected under neutrality, and analyses of both genotypes and phenotypes indicate that hybrids are rare in the hybrid zone. Rarity of hybrids indicates assortative mating and/or some form of fitness reduction in hybrids, which might maintain the species complex despite close genetic distance and introgression. These findings further support the treatment of the three taxa as distinct species.     

Comparative Assessment of Genetic and Morphological Variation at an Extensive Hybrid Zone between Two Wild Cats in Southern Brazil

Increased attention towards the Neotropical cats Leopardus guttulus and L. geoffroyi was prompted after genetic studies identified the occurrence of extensive hybridization between them at their geographic contact zone in southern Brazil. This is a region where two biomes intersect, each of which is associated with one of the hybridizing species (Atlantic Forest with L. guttulus and Pampas with L. geoffroyi). In this study, we conducted in-depth analyses of multiple molecular markers aiming to characterize the magnitude and spatial structure of this hybrid zone. We also performed a morphological assessment of these species, aiming to test their phenotypic differentiation at the contact zone, as well as the correlation between morphological features and the admixture status of the individuals. We found strong evidence for extensive and complex hybridization, with at least 40% of the individuals sampled in Rio Grande do Sul state (southernmost Brazil) identified as hybrids resulting from post-F1 generations. Despite such a high level of hybridization, samples collected in this state still comprised two recognizable clusters (genetically and morphologically). Genetically pure individuals were sampled mainly in regions farther from the contact zone, while hybrids concentrated in a central region (exactly at the interface between the two biomes). The morphological data set also revealed a strong spatial structure, which was correlated with the molecular results but displayed an even more marked separation between the clusters. Hybrids often did not present intermediate body sizes and could not be clearly distinguished morphologically from the parental forms. This observation suggests that some selective pressure may be acting on the hybrids, limiting their dispersal away from the hybrid zone and perhaps favoring genomic combinations that maintain adaptive phenotypic features of one or the other parental species.     

GENETIC STRUCTURE AND EVOLUTION OF A FIRE ANT HYBRID ZONE

Two introduced fire ants, Solenopsis invicta and S. richteri, hybridize over an extensive area in the United States spanning central Mississippi, Alabama, and western Georgia. We studied a portion of this hybrid zone in northwestern Mississippi in detail by sampling ants at many sites along two transects extending across the zone and examining gene frequency and size distributions at a large number of genetic and morphological markers. The distributional patterns at these markers are most consistent with the mosaic hybrid zone model, whereby the distribution of various fire ant genotypes is determined initially by the historical patterns of colonization of newly available habitats. However, these distributional patterns probably do not reflect the equilibrium state of interactions because of the very recent secondary contact of the species (< 60 yr) and the dynamic nature of available nesting habitats in this area. Our data suggest that, with prolonged contact and interaction, differential fitness of various hybrid genotypes due to intrinsic and extrinsic selective factors is important in structuring the hybrid zone. For instance, consistent differential introgression of morphological and genetic markers, combined with previous evidence of differences in developmental stability among genotypes, suggest reduced fitness of hybrids relative to parentals due to intrinsic selection (as may be caused by breakup of parental gene complexes). Furthermore, marked reductions in the occurrence of parental-like hybrids in areas where the similar parental species is common suggest reduced fitness of these parental-like hybrids in competition with the parentals (i.e., extrinsic selection). Because the relative roles of such deterministic as well as stochastic forces apparently vary both spatially and temporally, the eventual distribution of the various fire ant genotypes and the fate of the hybrid zone in the United States is difficult to predict.     

Hybridization and population structure of the Culex pipiens complex in the islands of Macaronesia

The Culex pipiens complex includes two widespread mosquito vector species, Cx. pipiens and Cx. quinquefasciatus. The distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. However, their distribution range overlaps in certain areas and interspecific hybridization has been documented. Genetic introgression between these species may have epidemiological repercussions for West Nile virus (WNV) transmission. Bayesian clustering analysis based on multilocus genotypes of 12 microsatellites was used to determine levels of hybridization between these two species in Macaronesian islands, the only contact zone described in West Africa. The distribution of the two species reflects both the islands' biogeography and historical aspects of human colonization. Madeira Island displayed a homogenous population of Cx. pipiens, whereas Cape Verde showed a more intriguing scenario with extensive hybridization. In the islands of Brava and Santiago, only Cx. quinquefasciatus was found, while in Fogo and Maio high hybrid rates (～40%) between the two species were detected. Within the admixed populations, second-generation hybrids (～50%) were identified suggesting a lack of isolation mechanisms. The observed levels of hybridization may locally potentiate the transmission to humans of zoonotic arboviruses such as WNV.     

Cytonuclear genetic architecture in mosquitofish populations and the possible roles of introgressive hybridization

Spatial genetic structure in populations of mosquitofish ( Gambusia ) sampled throughout the south-eastern United States was characterized using mitochondrial (mt) DNA and allozyme markers. Both sets of data revealed a pronounced genetic discontinuity (along a broad path extending from south-eastern Mississippi to north-eastern Georgia) that corresponds to a recently recognized distinction between the nominal forms G. affinis to the west and G. holbrooki to the east. However, several populations from the general contact region exhibited unusual allelic associations in high frequency, suggestive of evolutionary processes within a zone of introgressive hybridization. These involve: (i) cytonuclear profiles representing combinations of nuclear and mitochondrial genotypes that tended to be more nearly species-specific and concordant elsewhere; and (ii) significant nuclear gametic disequilibria, perhaps attributable to positive assortative mating and/or differential fitnesses of homospecific vs. recombinant genotypes. However, outside this suspected hybrid region, 'heterospecific' genetic markers also appeared in low frequency, thus complicating interpretations. These discordant alleles on a broader geographic scale may reflect: (a) the retention of polymorphisms from an ancestral gene pool; (b) occasional evolutionary convergence (especially with respect to electrophoretic mobility of allozyme alleles); (c) the 'footprints' of a moving hybrid zone; or (d) differential introgressive penetrance across the current hybrid region.     

Microgeographic population genetic structure in the northern water snake, Nerodia sipedon sipedon detected using microsatellite DNA loci

We describe the isolation and genetic characterization of eight microsatellite DNA loci from the northern water snake, Nerodia sipedon sipedon and use these loci to analyse levels of genetic differentiation between local (< 2 km apart) populations of these snakes in Ontario. These loci are variable, with expected heterozygosities ranging from 0.28 to 0.91, and can correctly exclude nonsires in parentage analyses with a high probability (0.998). Population analyses reveal significant deviation from expected heterozygosity levels for one population, probably a result of a null allele(s) at a single locus and small but significant levels of genetic differentiation among all three populations. This demonstrates that microgeographic genetic structure exists in this species, possibly due to limited dispersal.     

Effects of hybridization on the immunity of collared Ficedula albicollis and pied flycatchers F. hypoleuca, and their infection by haemosporidians

Because they are ubiquitous and typically reduce the fitness of hosts, parasites may play important roles in hybrid zone dynamics. Despite much work on herbivores and hybrid plants, the effect of parasites on the fitness of animal hybrids is poorly known. In an attempt to partly fill this gap, we examined the prevalence of avian haemosporidians Haemoproteus in a hybrid zone between collared Ficedula albicollis and pied flycatchers F. hypoleuca . 40 species-informative genetic markers allowed us to identify F1 hybrids, thus avoiding problems inherent in many studies that group hybrid genotypes. Furthermore, naturally occurring extra-pair paternity allowed us to test the immune responses of pure and hybrid nestlings to a novel antigen (phytohaemagglutinin) in a shared environment. In contrast to previous suggestions that animal hybrids may more often display resistance against parasites than plant hybrids, F1 hybrids exhibited prevalence of parasitism and immune responses that were intermediate between the two parental species. We also detected differences between the two parental species in their prevalence of infection, with the competitively dominant species (collared flycatcher) being less often infected by Haemoproteus . Overall, our results contribute to other recent data supporting the idea that the resistance of animals to parasites is variously and unpredictably affected by hybridization, and that there is a concordance in the general patterns observed in plants and animals. Haemosporidians in avian hybrids provide a useful system for investigating the interactions between hosts and parasites that characterize host contact zones.     

Assessing the genetic landscape of a contact zone: the case of European hare in northeastern Greece

The European hare populations of the Balkan Peninsula comprise two divergent phylogenetic lineages with discrete geographical distribution slightly overlapping in the area of northeastern Greece and Bulgaria. Here we elucidate their contact zone, by defining the spatial distributional pattern of the two highly divergent groups, detecting individuals of hybrid origin, and identifying genetic barriers present in the area of their co-existence. Specimens from northeastern Greece were assayed for lineage assignment and population genetic inference based on a 511 bp fragment of mitochondrial DNA control region and allelic data from 10 microsatellite loci. Bayesian analyses on original and simulated genotypes were performed allowing for the contact zone delineation. Our results indicate high genetic diversity in both nuclear and mitochondrial DNA, strong population structure and non random spatial distribution of the differentiated gene pools. The information provided by the two types of molecular markers yielded consistent results. This study comprises a fine scale analysis of the contact zone between the two evolutionary lineages of European brown hares in northeastern Greece. Specific questions on the spatial patterns where addressed for the first time. Furthermore, hypotheses regarding the presence of hybrids were also tested. As a result, interpretive power to the diversity patterns observed today in the Balkans was added and previously overlooked aspects of the species biology were highlighted.     

Interspecific gene flow in a multispecies oak hybrid zone in the Sierra Tarahumara of Mexico

Background and Aims

Interspecific gene flow can occur in many combinations among species within the genus Quercus, but simultaneous hybridization among more than two species has been rarely analysed. The present study addresses the genetic structure and morphological variation in a triple hybrid zone formed by Q. hypoleucoides, Q. scytophylla and Q. sideroxyla in north-western Mexico.

Methods

A total of 247 trees from ten reference and 13 presumed intermediate populations were characterized using leaf shape variation and geometric morphometrics, and seven nuclear microsatellites as genetic markers. Discriminant function analysis was performed for leaf shape variation, and estimates of genetic diversity and structure, and individual Bayesian genetic assignments were obtained.

Key Results

Reference populations formed three completely distinct groups according to discriminant function analysis based on the morphological data, and showed low, but significant, genetic differentiation. Populations from the zone of contact contained individuals morphologically intermediate between pairs of species in different combinations, or even among the three species. The Bayesian admixture analysis found that three main genetic clusters best fitted the data, with good correspondence of reference populations of each species to one of the genetic clusters, but various degrees of admixture evidenced in populations from the contact area.

Conclusions

The three oak species have formed a complex hybrid zone that is geographically structured as a mosaic, and comprising a wide range of genotypes, including hybrids between different species pairs, backcrosses and probable triple hybrids.     

Unexpected hybridization patterns in Near Eastern terrapins (Mauremys caspica,M. rivulata) indicate ancient gene flow across the Fertile Crescent

Recent studies indicate that hybridization in animals occurs more frequently than previously thought and that it may play an important evolutionary role. Chelonians are capable of extensive hybridization, raising the question how chelonian species evolve and maintain genetic integrity despite hybridization. Here, we use two sister species with parapatric distribution, Mauremys caspica and M. rivulata, as our model. These taxa are estimated to have diverged some 5.3–7.0 million years ago. Using rangewide sampling and 13 unlinked polymorphic microsatellite markers, five nuclear loci and one mitochondrial gene, we show that hybridization is rare along the contact zone of the two species in Turkey. However, we discovered an unexpected hybrid swarm in the southern Levant that has been hitherto identified with M. rivulata. This hybrid swarm is separated from the inland species M. caspica by a 700‐km‐wide distribution gap corresponding to the Syrian Desert. Ecological palaeomodelling suggests that during more humid climatic episodes in the Last Glacial Maximum and mid‐Holocene, the current contact zone extended into the southern Levant, facilitating the establishment of the now isolated hybrid swarm. Our results support that there is not necessarily a general hybridization pattern in a given species couple and that the extent of gene flow may differ considerably in different parts of the distribution range. Moreover, our results highlight that studies on hybridization should not focus only on extant contact and hybrid zones, but should use rangewide sampling to detect signals of ancient hybridization that might otherwise be missed.     

BIOCHEMICAL PHENOTYPIC AND GENETIC STUDIES OF TWO INTRODUCED FIRE ANTS AND THEIR HYBRID (HYMENOPTERA: FORMICIDAE)

Two introduced fire ants, Solenopsis invicta and S. richteri, and their hybrid were studied using phenotypic markers from gas chromatographic analysis of hydrocarbons and venom alkaloids, as well as genetic markers from enzyme electrophoresis. Both methods show that extensive gene introgression is occurring over a distance of at least 120 km at the contact zone between the two forms in eastern Mississippi. Genetic analyses suggest that the hybrid population does not depart dramatically from panmixia. Also, recombinant genotypes predominate in the hybrid zone, indicating that F₁ hybrids are viable. Allele frequency clines through the hybrid zone are apparent for four polymorphic loci. Data sets generated by the chromatographic and electrophoretic methods are highly concordant in that they differentiate completely between the two forms and agree in designating colonies from the contact zone as hybrid or parental in a high proportion (90%) of cases. The two methods can serve as complementary tools for studying closely related but genetically distinct populations in this, and perhaps other, groups of insects.     

Extensive hybridization in a contact zone between MacGillivray's warblers Oporornistolmiei and mourning warblers O. philadelphia detected using molecular and morphological analyses

There are many pairs of related western and eastern avian taxa in North America, and for many of these, little is known about their interactions in sympatry. One example is provided by MacGillivray's warblers Oporornis tolmiei and mourning warblers Oporornis philadelphia . There have been occasional reports of range contact and hybridization between these forms, but recent authors have doubted these reports. We show that these two species do in fact come into extensive range contact in the southern Peace Region of British Columbia, just east of the Rocky Mountains. We analyze whether patterns of variation in morphometric traits, eye-arcs, a mitochondrial DNA marker (COI), and a Z-chromosome marker (CHD1Z) are consistent with reproductive isolation or hybridization in this contact zone. Each trait shows strong differences between allopatric MacGillivray's warblers and allopatric mourning warblers, yet in the contact zone there are many birds with a combination of traits typical of both species. This is clearly seen in the molecular markers, for which 18 of 50 birds genotyped in the contact zone have both western and eastern alleles. These patterns strongly indicate the presence of an extensive hybrid zone between MacGillivray's and mourning warblers. Variation in each of the four traits is explained well by a single sigmoidal cline, with a width of roughly 150  km (or 130  km based only on the molecular markers). This is only the fourth hybrid zone known among North American wood-warblers (Parulidae).     

Hybridization between two gartersnake species (<Emphasis Type="Italic">Thamnophis</Emphasis>) of conservation concern: a threat or an important natural interaction?

Distinguishing between hybrid zones formed by secondary contact versus parapatric divergence-with-gene-flow is an important challenge for understanding the interplay of geographic isolation and local adaptation in the origin of species. Similarly, distinguishing between natural hybrid zones and those that formed as a consequence of recent human activities has important conservation implications. Recent work has demonstrated the existence of a narrow hybrid zone between the plains gartersnake (Thamnophis radix) and Butler’s gartersnake (T. butleri) in the Great Lakes region of North America, raising questions about the history and conservation value of genetically admixed populations. Both taxa are of conservation concern, and it is not clear whether to regard hybridization as a threat or a natural interaction. Here we use phylogeographic and population genetic methods to assess the timescales of divergence and hybridization, and test for evidence that the hybrid zone is of recent origin. We assayed AFLP markers and ND2 mitochondrial DNA (mtDNA) sequences from T. radix, T. butleri, and the closely related short-headed gartersnake (T. brachystoma) throughout their North American ranges. We find shallow mtDNA divergence overall and high levels of variation within the contact zone. These patterns are inconsistent with recent contact of long-diverged taxa. It is not possible to distinguish true divergence-with-gene-flow from a long-term secondary contact zone, but we infer that the hybrid zone is a long-standing, natural interaction.     

THE VARIABLE GENOMIC ARCHITECTURE OF ISOLATION BETWEEN HYBRIDIZING SPECIES OF HOUSE MICE

Studies of the genetics of hybrid zones can provide insight into the genomic architecture of species boundaries. By examining patterns of introgression of multiple loci across a hybrid zone, it may be possible to identify regions of the genome that have experienced selection. Here, we present a comparison of introgression in two replicate transects through the house mouse hybrid zone through central Europe, using data from 41 single nucleotide markers. Using both genomic and geographic clines, we found many differences in patterns of introgression between the two transects, as well as some similarities. We found that many loci may have experienced the effects of selection at linked sites, including selection against hybrid genotypes, as well as positive selection in the form of genotypes introgressed into a foreign genetic background. We also found many positive associations of conspecific alleles among unlinked markers, which could be caused by epistatic interactions. Different patterns of introgression in the two transects highlight the challenge of using hybrid zones to identify genes underlying isolation and raise the possibility that the genetic basis of isolation between these species may be dependent on the local population genetic make-up or the local ecological setting.     

Inter-species hybridization among Neotropical cats of the genus Leopardus, and evidence for an introgressive hybrid zone between L. geoffroyi and L. tigrinus in southern Brazil

Natural hybrid zones between distinct species have been reported for many taxa, but so far, few examples involve carnivores or Neotropical mammals in general. In this study, we employed mitochondrial DNA (mtDNA) sequences and nine microsatellite loci to identify and characterize a hybrid zone between two Neotropical felids, Leopardus geoffroyi and L. tigrinus, both of which are well-established species having diverged from each other c. 1 million years ago. These two felids are mostly allopatric throughout their ranges in South America, with a narrow contact zone that includes southern Brazil. We present strong evidence for the occurrence of hybridization between these species and identify at least 14 individuals (most of them originating from the geographical contact zone) exhibiting signs of interspecific genomic introgression. The genetic structure of Brazilian L. tigrinus populations seems to be affected by this introgression process, showing a gradient of differentiation from L. geoffroyi correlated with distance from the contact zone. We also corroborate and extend previous findings of hybridization between L. tigrinus and a third related felid, L. colocolo, leading to an unusual situation for a mammal, in which the former species contains introgressed mtDNA lineages from two distinct taxa in addition to its own.