Premating barriers to gene exchange and their implications for the structure of a mosaic hybrid zone between Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae)

Many hybrid zones contain a deficit of hybrid genotypes relative to expectations from tension zone models. This is often associated with separation of parental genotypes into distinct habitats (mosaicism), but sometimes parentals can be found co-occuring in the same local population (bimodality). In both cases, prezygotic isolation may play an important role in determining the genotypic composition of the zone. Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae) meet and form a complex hybrid zone in northern Spain. Analysis of stridulatory peg numbers reveals partial spatial and seasonal isolation in a 25 km2 area of the zone: C. jacobsi phenotypes predominate in June and July and are present in both valley and mountain habitats; C. brunneus phenotypes predominate in August and are restricted to valley habitats, always in sympatry with C. jacobsi. Strong assortative mating was observed in laboratory mating experiments. Spatial, seasonal and behavioural isolation combine to produce strong premating isolation in the study area. These results suggest a role for premating isolation in maintaining both the mosaic structure and bimodality of this hybrid zone.     

Assortative preferences and discrimination by females against hybrid male song in the grasshoppers Chorthippus brunneus and Chorthippus jacobsi (Orthoptera: Acrididae)

The grasshoppers Chorthippus brunneus and Chorthippus jacobsi are highly differentiated for male mating signals, and form a mosaic hybrid zone in northern Spain. At some sites within this zone, many hybrids are observed. At others, few hybrids are observed. Such bimodal sites may reflect recent contacts between parental genotypes, or local variation in levels of assortative mating or selection against hybrids. Playback of 12 parental and F1 male songs to 296 parental and hybrid females revealed positive assortative preferences in C. brunneus and C. jacobsi females, supporting a direct role of male mating signals in female choice. However, all female genotypic classes showed reduced responsiveness to F1 male songs. Such sexual selection against hybrids is consistent with the narrow cline width observed in the field for song characters relative to morphology. These results have implications for the genetic structure of the hybrid zone and for models of speciation by reinforcement.     

Hybrids between Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae) do not show endogenous postzygotic isolation

Studies of prezygotic and postzygotic isolation in Drosophila have shown in general that species in sympatry tend to evolve prezygotic barriers earlier than do species in allopatry. However, postzygotic barriers tend to evolve at the same evolutionary rate in both sympatric and allopatric species. In contrast to these observations, the grasshoppers Chorthippus parallelus parallelus and C. p. erythropus show complete hybrid male sterility but only limited prezygotic isolation after an estimated 0.5 millions years of divergence. Like their congeners, C. brunneus and C. jacobsi form a hybrid zone where their ranges meet in northern Spain. However, the hybrid zone is mosaic and bimodal and, in contrast to the high levels of postzygotic isolation between C. parallelus subspecies, these two species showed no significant reduction in hybrid fitness in F₁ or backcross generations relative to the parental generations. The level of prezygotic isolation in laboratory tests was comparable to that between C. parallelus subspecies. These results suggest that endogenous postzygotic isolation does not play an important role in the reproductive isolation between C. brunneus and C. jacobsi , or in determining the structure of the hybrid zone. Exogenous postzygotic isolation may be present and should be tested in future studies.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 195–203.     

SPATIAL STRUCTURE AND HABITAT VARIATION IN A GRASSHOPPER HYBRID ZONE

Abstract A hybrid zone between the grasshoppers Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae) in northern Spain has been analyzed for variation in morphology and ecology. These species are readily distinguished by the number of stridulatory pegs on the hind femur. Both sexes are fully winged and inhabit disturbed habitats throughout the study area. We develop a maximum-likelihood approach to fitting a two-dimensional cline to geographical variation in quantitative traits and for estimating associations of population mean with local habitat. This method reveals a cline in peg number approximately 30 km south of the Picos de Europa Mountains that shows substantial deviations in population mean compared with the expectations of simple tension zone models. The inclusion of variation in local vegetation in the model explains a significant proportion of the residual variation in peg number, indicating that habitat-genotype associations contribute to the observed spatial pattern. However, this association is weak, and a number of populations continue to show strong deviations in mean even after habitat is included in the final model. These outliers may be the result of long-distance colonization of sites distant from the cline center or may be due to a patchy pattern of initial contact during postglacial expansion. As well as contrasting with the smooth hybrid zones described for Chorthippus parallelus , this situation also contrasts with the mosaic hybrid zones observed in Gryllus crickets and in parts of the hybrid zone between Bombina toad species, where habitat-genotype associations account for substantial amounts of among-site variation.     

Bimodal hybrid zones and speciation

Contact zones exemplify a series of stages in speciation. In unimodal hybrid zones intermediates predominate; in bimodal zones hybrids are rare and parental forms predominate; and finally, species might overlap, but never hybridize. Recent studies show bimodality to be associated strongly with assortative mating or fertilization, and only weakly with overall levels of genetic divergence or intrinsic genomic incompatibility. Ecological divergence across most bimodal hybrid zones suggests that ecology contributes more to speciation than genomic incompatibility. This continuum of stable contact zones provides empirical evidence for a route to speciation, which does not require allopatry.     

Temperature cycles induce a bimodal activity pattern in ruin lizards: masking or clock-controlled event? A seasonal problem.

The daily locomotor activity pattern of Ruin lizards in the field is mainly unimodal, except for summer months when soil temperatures exceed 40 degrees C to 42 degrees C around midday. In such a situation, lizards reduce their locomotor activity around midday to avoid overheating, and thus their daily activity pattern becomes bimodal. The bimodal pattern expressed in the field is usually retained in the free-running rhythm under constant temperature and DD for a couple of weeks, after which the bimodal pattern changes into a unimodal pattern. In the present study, the authors examined whether 24-h temperature cycles (TCs) would change lizard activity from a unimodal to a bimodal pattern. Administration of TCs to unimodal lizards free-running in DD is able to entrain locomotor rhythms and to induce a bimodal pattern both in summer and autumn-winter. There are, however, striking seasonal differences in the effectiveness with which TCs achieve bimodality: (a) Numbers of lizards rendered bimodal are significantly higher in summer than in autumn-winter; (b) TCs require less time to achieve bimodality in summer than autumn-winter; (c) bimodality is retained as an aftereffect in the postentrainment free-run in summer, but not in autumn-winter; (d) TCs change activity duration in summer, but not in autumn-winter. All this demonstrates the existence of seasonal changes in responsiveness of the circadian oscillators controlling activity to the external factors inducing bimodality. Oscillators' responsiveness is high in summer, when bimodality is the survival strategy of Ruin lizards to avoid overheating around midday in open fields, and low in autumn-winter, when bimodality has no recognizable adaptive significance.     

Mating signal variation and bimodality in a mosaic hybrid zone between Chorthippus grasshopper species

The grasshoppers Chorthippus brunneus and Chorthippus jacobsi are easily distinguished by male calling song and the number of stridulatory pegs on the hind femur, and form a mosaic hybrid zone in northern Spain. In this paper, we fit a two-dimensional cline to variation in male calling song characters, which are of particular interest as they are likely to be involved in mate choice by females. As with variation in peg number, local habitat makes only a small contribution in explaining deviations in mean song score from clinal expectations. However, the fitted width of the cline for song characters is significantly narrower than for peg number, suggesting that mating signals may be associated with reduced hybrid fitness in the field and that recombination rates are sufficient to allow clines for different characters to diverge in width. Despite this, estimates for the overall elevation in linkage disequilibrium at the zone center, based on covariance between peg and song characters, reveal a substantial overrepresentation of parental genotypes at the cline center relative to the expectations of a tension zone of similar width. Examination of covariance at individual sites reveals that this inflated estimate of linkage disequilibrium is caused by several sites where the distribution of phenotypes is effectively bimodal. This substantial variation in linkage disequilibrium at the cline center could result from local variation in the strength of assortative mating or selection against hybrids, or may reflect the long-distance colonization of empty habitat from outside the hybrid zone, which would continually create new contacts between parental genotypes at the cline center. Hybrid zones like this, in which strong linkage disequilibrium occurs in some situations but not in others, are of particular relevance to speciation research and allow investigation of the spread of combinations of alleles through different genetic and ecological backgrounds.     

Genetic differentiation and habitat connectivity across towhee hybrid zones in Mexico

Hybridization can either reinforce or erode species boundaries; therefore, hybrid zones offer a natural experimental setting in which to assess the dynamics of reproductive isolation. Secondary contact zones, in particular, present a partial separation of the original divergence mechanisms and the subsequent genomic architecture of reproductive isolation (or lack thereof). The spatial context of secondary contact and its consequent effect on dispersal play vital roles on the contact’s outcome. In a hybrid complex between two towhee species in Mexico, Pipilo maculatus and Pipilo ocai, two major hybrid gradients provide natural replicates for comparison. However, genetic analyses demonstrate significant divergence between geographically separate parental populations of each species and divergence of populations within each hybrid zone. The two hybrid transects (Teziutlán and Transvolcanic) are distinct and evidence suggests allelic introgression both across the species boundary and between the two transects. Habitat corridors for dispersal represent functional connectivity hotspots where the two transects meet. Both habitat connectivity and genetic differentiation between geographically disparate parental populations appear to influence the dynamics of gene flow across the hybrid gradients. In southern sympatric populations (Mt. Orizaba and Oaxaca) where morphological evidence for hybridization is scarce, opposing species’ alleles appear to traverse through the hybrid zones rather than arising from cryptic local hybridization. These results illustrate the importance of environmentally mediated gene flow in the context of secondary contact as an important force influencing evolutionary trajectory.     

The modality of nine Triturus newt hybrid zones assessed with nuclear,mitochondrial and morphological data

The shape of hybrid zones provides insight into genetic isolation between the evolutionary lineages involved: the greater the limitation to gene flow, the farther along the unimodal to bimodal continuum. We study hybrid zone modality in a group of closely related species at a variety of levels of genetic divergence. We explore the degree and the geography of hybridization in 12 transects of Triturus newts, for nine of the ten secondary contact zones in Europe, using nuclear and mitochondrial encoded genetic data and morphological characteristics. Species status is verified by examining the correlation between the three marker systems in a geographical context. At 40 allozyme loci, two transects classify as unimodal, three as bimodal and two as intermediate, while the signal for hybridization is weak or absent in five transects. One zone studied in duplicate was classified as intermediate in one region and as bimodal in another region. mtDNA introgression is frequent and extends beyond nuclear introgression in two transects. Morphology provides additional evidence for hybridity, including one transect for which the signal of nuclear gene flow is weak. Compared to simulations allowing panmixia, the observed allozyme transitions at contact zones show a deficit of backcrossing to various degrees. Over all transects, there is a weak negative relationship between the level of hybridization and allozyme genetic distance for species pairs, consistent with Bateson–Dobzhansky–Muller effects. This observation, based upon highly comparable data for a single genus, supports conclusions derived from analyses over a wide variety of other taxa. © 2014 The Authors. Biological Journal of the Linnean Society published by John Wiley & Sons Ltd on behalf of The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 604–622.     

Geographic variation in the structure of oak hybrid zones provides insights into the dynamics of speciation

Studying geographic variation in the rate of hybridization between closely related species could provide a useful window on the evolution of reproductive isolation. Reinforcement theory predicts greater prezygotic isolation in areas of prolonged contact between recently diverged species than in areas of recent contact, which implies that old contact zones would be dominated by parental phenotypes with few hybrids (bimodal hybrid zones), whereas recent contact zones would be characterized by hybrid swarms (unimodal hybrid zones). Here, we investigate how the hybrid zones of two closely related Chinese oaks, Quercus mongolica and Q. liaotungensis, are structured geographically using both nuclear and chloroplast markers. We found that populations of Q. liaotungensis located around the Changbai Mountains in Northeast China, an inferred glacial refugium, were introgressed by genes from Q. mongolica, suggesting historical contact between the two species in this region. However, these introgressed populations form sharp bimodal hybrid zones with Q. mongolica. In contrast, populations of Q. liaotungensis located in North China, which show no sign of ancient introgression with Q. mongolica, form unimodal hybrid zones with Q. mongolica. These results are consistent with the hypothesis that selection against hybrids has had sufficient time to reinforce the reproductive barriers between Q. liaotungensis and Q. mongolica in Northeast China but not in North China.     

Distinctiveness in the face of gene flow: hybridization between specialist and generalist gartersnakes

Patterns of divergence and polymorphism across hybrid zones can provide important clues as to their origin and maintenance. Unimodal hybrid zones or hybrid swarms are composed predominantly of recombinant individuals whose genomes are patchworks of alleles derived from each parental lineage. In contrast, bimodal hybrid zones contain few identifiable hybrids; most individuals fall within distinct genetic clusters. Distinguishing between hybrid swarms and bimodal hybrid zones can be important for taxonomic and conservation decisions regarding the status and value of hybrid populations. In addition, the causes of bimodality are important in understanding the generation and maintenance of biological diversity. For example, are distinct clusters mostly reproductively isolated and co‐adapted gene complexes, or can distinctiveness be maintained by a few ‘genomic islands’ despite rampant gene flow across much of the genome? Here we focus on three patterns of distinctiveness in the face of gene flow between gartersnake taxa in the Great Lakes region of North America. Bimodality, the persistence of distinct clusters of genotypes, requires strong barriers to gene flow and supports recognition of distinct specialist (Thamnophis butleri) and generalist (Thamnophis radix) taxa. Concordance of DNA‐based clusters with morphometrics supports the hypothesis that trophic morphology is a key component of divergence. Finally, disparity in the level of differentiation across molecular markers (amplified fragment length polymorphisms) indicates that distinctiveness is maintained by strong selection on a few traits despite high gene flow currently or in the recent past.     

A dispersal-dependent zone of introgressive hybridization between weakfish, Cynoscion regalis, and sand seatrout, C. arenarius, (Sciaenidae) in the Florida Atlantic

Five diagnostic codominant nuclear DNA markers and a diagnostic mitochondrial DNA marker were used to survey weakfish (Cynoscion regalis) and sand seatrout (C. arenarius), with particular focus on heretofore uncharacterized juvenile populations along the Florida (FL) Atlantic coast. Geographic and reproductive ranges of weakfish and sand seatrout were shown to overlap on the Atlantic coast along north and central FL. An active bidirectional zone of introgressive hybridization exists between these taxa, centered in the St Johns River, FL. Strong patterns of Hardy-Weinberg, linkage, and cytonuclear disequilibrium and a bimodal hybrid index distribution were observed for juvenile cohorts in the zone center, coupled with narrow (～240 km) concordant clines. Parental forms had disparate habitat preferences; hybrid forms occurred predominantly in intermediate habitats. All genetic data were consistent with the hypothesis that the C. arenarius-C. regalis hybrid zone is maintained by a dynamic equilibrium between continued interspecific gene flow and one or more opposing forces. Cytonuclear analyses indicated that parental forms mate assortatively in the zone but that mate recognition was imperfect. Ethological mating dynamics are likely stabilized by some form of endogenous or exogenous postfertilization selection against hybrids such that parental taxa will likely continue to evolve independently.     

Measuring dispersal and detecting departures from a random walk model in a grasshopper hybrid zone

Abstract. 1. The grasshopper species Chorthippus brunneus and C. jacobsi form a complex mosaic hybrid zone in northern Spain. Two mark–release–recapture studies were carried out near the centre of the zone in order to make direct estimates of lifetime dispersal.
2. A model framework based on a simple random walk in homogeneous habitat was extended to include the estimation of philopatry and flying propensity. Each model was compared with the real data, correcting for spatial and temporal biases in the data sets.
3. All four data sets (males and females at each site) deviated significantly from a random walk. Three of the data sets showed strong philopatry and three had a long dispersal tail, indicating a low propensity to move further than predicted by the random walk model.
4. Neighbourhood size estimates were 76 and 227 for the two sites. These estimates may underestimate effective population size, which could be increased by the long tail to the dispersal function. The random walk model overestimates lifetime dispersal and hence the minimum spatial scale of adaptation.
5. Best estimates of lifetime dispersal distance of 7–33 m per generation were considerably lower than a previous indirect estimate of 1344 m per generation. This discrepancy could be influenced by prezygotic isolation, an inherent by-product of mosaic hybrid zone structure.     

The size structure of lacustrine Arctic charr (Pisces: Salmonidae) populations

The size structure of Arctic charr (Salvelinus alpinus) populations was analysed using published information; 44% of populations were bimodal, with both large (normal) and small (dwarf) morphs occurring within a cohort. The remaining populations were unimodal, consisting of normal-sized or of stunted adults. Bimodal populations increased in frequency with latitude and were characteristic of large, deep lakes with few fish species. The age and size at which bimodality developed, the size difference between the morphs, and the frequency of cannibalism in charr populations increased with latitude. A variety of genotypic and phenotypic explanations are examined. The evidence for specific differences between the morphs is unconvincing. Various competition and predation hypotheses fail to explain the occurrence of bimodality. Cannibalism does not cause bimodality despite being strongly associated with it since bimodality develops before charr become cannibalistic. Much of the variation in charr size structure is suggested to be a consequence of increased seasonality in food supply in more northerly environments, coupled with feeding size thresholds. The latter result in larger members of a cohort being able to maintain growth rates on seasonally abundant prey while smaller individuals which cannot catch these items form a second mode of more slowly growing fish. Bimodality is documented in a number of other, predominantly northern, fish species.     

Seasonal precipitation and continentality drive bimodal growth in Mediterranean forests

Tree phenology is sensitive to climate warming and changes in seasonal precipitation. Long xylogenesis records are scarce, thus limiting our ability to analyse how radial growth responds to climate variability. Alternatively, process-based growth models can be used to simulate intra-annual growth dynamics and to better understand why growth bimodality varies along temperature and precipitation gradients. We used the Vaganov-Shashkin (VS) growth model to analyse the main climatic drivers of growth bimodality in eight trees and shrubs conifers (four pines and four junipers) across Spain. We selected eleven sites with different continentality degree and spring/autumn precipitation ratios since we expected to find pronounced bimodal growth in less continental sites with spring and autumn precipitation peaks. The VS model successfully simulated annual growth rates at all sites as a function of daily temperature and soil moisture data. Bimodal growth patterns clustered into less continental sites showing low spring/autumn precipitation ratios. This finding agrees with observed climate-growth associations showing that growth was enhanced by wet-cool winter-to-spring conditions, but also by wet autumn conditions in the most bimodal sites. We observed a stronger growth bimodality in pines compared to junipers. We discuss the spatial variability of climate drivers in bimodality growth pattern and how increasing continentality and shifts in seasonal precipitation could affect growth patterns. Bimodality could be an advantageous response to overcome summer drought in Mediterranean forests. The ability of some species to reactivate growth during autumn might determine their capacity to withstand increasing summer aridity.     

Species pools and the "hump-back" model of plant species diversity: an empirical analysis at a relevant spatial scale

We investigated the relative roles of productivity, the species pool, and spatial habitat structure in determining local species richness (alpha diversity) of plant communities within a single, well-defined landscape unit, at spatial and ecological scales where the relationship between community productivity and species diversity often assumes a unimodal or "hump-back" form. At high levels of productivity, the decrease-phase of the unimodal model of the diversity-productivity relationship is typically explained as the dynamic outcome of increased competitive exclusion, but it may also be the passive consequence of a small pool of species possessing attributes necessary to competitively survive in high-fertility environments. We conducted statistical analyses of previously collected data to determine whether variations in local richness in the herbaceous vegetation of a Slovakian mountain valley were best explained by habitat productivity itself (which presumably leads to more intense competition) or by the sizes of the relevant community species pools. We also used measures of spatial habitat structure to investigate the extent to which habitat patchiness influenced patterns of species diversity. In the study system, both community biomass and size of the species pools contributed significantly to local species richness, but the positive effect of the species pools was about twice as important as the negative effect of biomass. The combined area of related associations (alliance area), association perimeter, and habitat patch geometry were all closely related to species pool size.     

Possible human impacts on adaptive radiation: beak size bimodality in Darwin's finches

Adaptive radiation is facilitated by a rugged adaptive landscape, where fitness peaks correspond to trait values that enhance the use of distinct resources. Different species are thought to occupy the different peaks, with hybrids falling into low-fitness valleys between them. We hypothesize that human activities can smooth adaptive landscapes, increase hybrid fitness and hamper evolutionary diversification. We investigated this possibility by analysing beak size data for 1755 Geospiza fortis measured between 1964 and 2005 on the island of Santa Cruz, Galápagos. Some populations of this species can display a resource-based bimodality in beak size, which mirrors the greater beak size differences among species. We first show that an historically bimodal population at one site, Academy Bay, has lost this property in concert with a marked increase in local human population density. We next show that a nearby site with lower human impacts, El Garrapatero, currently manifests strong bimodality. This comparison suggests that bimodality can persist when human densities are low (Academy Bay in the past, El Garrapatero in the present), but not when they are high (Academy Bay in the present). Human activities may negatively impact diversification in 'young' adaptive radiations, perhaps by altering adaptive landscapes.     

Differential introgression across newt hybrid zones: Evidence from replicated transects

Genomic heterogeneity of divergence between hybridizing species may reflect heterogeneity of introgression, but also processes unrelated to hybridization. Heterogeneous introgression and its repeatability can be directly tested in natural hybrid zones by examining multiple transects. Here, we studied hybrid zones between the European newts Lissotriton montandoni and two lineages of Lissotriton vulgaris, with replicate transects within each zone. Over 1,000 nuclear genes located on a linkage map and mitochondrial DNA were investigated using geographical and genomic clines. Overall, the five transects were all similar, showing hallmarks of strong reproductive isolation: bimodal distribution of genotypes in central populations and narrow allele frequency clines. However, the extent of introgression differed between the zones, possibly as a consequence of their different ages, as suggested by the analysis of heterozygosity runs in diagnostic markers. In three transects genomic signatures of small‐scale (~2 km) zone movements were detected. We found limited overlap of cline outliers between transects, and only weak evidence of stronger differentiation of introgression between zones than between transects within zones. Introgression was heterogeneous across linkage groups, with patterns of heterogeneity similar between transects and zones. Predefined candidates for increased or reduced introgression exhibited only a subtle tendency in the expected direction, suggesting that interspecific differentiation is not a reliable indicator for the strength of introgression. These hierarchically sampled hybrid zones of apparently different ages show how introgression unfolds with time and offer an excellent opportunity to dissect the dynamics of hybridization and architecture of reproductive isolation at advanced stages of speciation.     

ELECTROPHORETIC CONFIRMATION OF INTERSPECIFIC HYBRIDIZATION IN AESCULUS (HIPPOCASTANACEAE) AND THE GENETIC STRUCTURE OF A BROAD HYBRID ZONE

Within a broad (>200 km wide) hybrid zone involving three parapatric species of Aesculus, we observed coincident clines in allele frequency for 6 of 14 electrophoretic loci. The cooccurrence of alleles characteristic of A. pavia, A. sylvatica, and A. flava was used to estimate genetic admixtures in 48 populations involving various hybrids between these taxa in the southeastern United States. High levels of allelic polymorphism (up to 40% greater than the parental taxa) were observed in hybrid populations and also in some populations bordering the hybrid zone. A detailed analysis of a portion of the hybrid zone involving A. pavia and A. sylvatica revealed a highly asymmetrical pattern of gene flow, predominantly from Coastal Plain populations of A. pavia into Piedmont populations of A. sylvatica. Computer simulations were used to generate expected genotypic arrays for parental, F₁; and backcross individuals, which were compared with natural populations using a character index scoring system. In these comparisons, hybrid individuals could be distinguished from either parent, but F₁ and backcross progeny could not be distinguished from each other. Most hybrid populations were found to include hybrids and one of the parental taxa, but never both parents. Three populations appeared to be predominantly hybrids with no identifiable parental individuals. Hybrids occurred commonly at least 150 km beyond the range of A. pavia, but usually not more than 25 km beyond the range of A. sylvatica. Introgression, suggested by genetically hybrid individuals and significant gene admixtures of two or more species in populations lacking morphological evidence of hybridization, may extend the hybrid zone further in both directions. The absence of one or both parental species from hybrid populations implies a selective disadvantage to parentals in the hybrid zone and/or that hybridization has occurred through long-distance gene flow via pollen, primarily from A. pavia into A. sylvatica. Long-distance pollen movement in plants may generate hybrid zones of qualitatively different structure than those observed in animals, where gene flow involves dispersal of individuals.     

Morphological variation in intergrade pupfish populations from the Pecos River, Texas, U.S.A.

In the early 1980s, sheepshead minnow Cyprinodon variegates was introduced into the Pecos River, Texas, U.S.A. where it hybridized with the endemic Pecos pupfish C. pecosensis . By 1985, pupfish populations throughout approximately 300 km of the river consisted exclusively of individuals of hybrid origin (intergrades). There was significant ( P <0·05) geographic variation in most morphological characters; the general pattern of variation was of a bidirectional cline centred near Pecos, Texas. At that site, morphology of intergrade populations resembled mostly that of the introduced species. Upstream and downstream from Pecos, morphology shifted progressively toward that typical of the native form. Intergrade populations were morphologically intermediate to the parental forms, showed a rapid approach to random assortment of characters, and generally exhibited greater morphological variability than occurred in either parent species. These observations and the consistent lack of bimodality in frequency distributions of a morphological hybrid index support the contention that intergrade populations comprise panmictic admixtures of C. variegates and C. pecosensis .