Phenotypic plasticity of larval retreat design in a net-spinning caddisfly

Larval Macrostemum carolina caddisflies construct silken catchnetswithin protective retreats, often on submerged trees and branches(i.e., snags). In the Savannah River, M. carolina larvae constructthree distinct retreats that differ in the configuration ofthe water entrance hole: (1) at the end of a silken tube, (2)flush with the top of the retreat, and (3) backed by a 180-degreesilken backstop. To identify the proximate mechanism mediatingthis retreat polymorphism, we removed larvae of known phenotypefrom their original retreats and brought them into the laboratory,allowing them to construct new retreats. We found that (1) larvaecan construct more than one type of retreat, so variation inthis behavior is not under strict genetic control; (2) larvaedo not preferentially reconstruct their original retreat design,so these alternative behaviors apparently exhibit little heritability;and (3) larvae primarily construct each phenotype in a particularmicrohabitat (i.e., "tube" and "backstop" retreats are principallyconstructed on the downstream half of the snag, and "flush"retreats on the upstream–bottom quadrant). Therefore,the retreat polymorphism in M. carolina is phenotypically plasticand is controlled by microhabitat location or a correlated environmentalvariable. Most net-spinning caddisflies construct their netsin fairly specific microhabitats. However, behavioral plasticityallows M. carolina larvae to construct retreats all around asnag, thereby reducing potentially intense competition for spacewith other net-spinning caddisflies. Consequently, this maybe the ultimate reason this polymorphism evolved.     

Postglacial genetic differentiation of reproductive ecotypes of kokanee Oncorhynchus nerka in Okanagan Lake, British Columbia

Okanagan Lake, south-central interior of BC, contains two reproductive ecotypes of kokanee Oncorhynchus nerka ; individuals spawn in tributary streams ('stream-spawners') as well as on shoreline gravel areas ('beach-spawners'). We tested the hypothesis that these sympatric ecotypes comprise a single panmictic population by assaying variation in morphological traits and at allozyme, mitochondrial and minisatellite DNA loci in fish collected from three stream-spawning and two beach-spawning sites. No morphological traits consistently distinguished the reproductive ecotypes with the exception of the number of anal fin rays which was greater in stream-spawning kokanee. Four of 18 allozyme loci screened were polymorphic, but no significant allele frequency differences were detected among populations within ecotypes or between ecotypes. Similarly, allele frequencies at two minisatellite DNA loci were not significantly different among populations or between ecotypes. By contrast, significant differences in the frequencies of mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) haplotypes were detected between stream- and beach-spawners, but not among populations within ecotypes. Further, two RFLPs that distinguished stream- and beach-spawning adults were found in juvenile kokanee sampled from the limnetic zone of Okanagan Lake. The two mtDNA RFLPs and a d-loop sequence variant appear to be unique to Okanagan Lake kokanee because we did not observe these haplotypes in sockeye salmon and kokanee sampled outside of Okanagan Lake. Our data suggest that: (i) there is restricted female-mediated gene flow between stream- and beach-spawning kokanee in Okanagan Lake, (ii) the forms have diverged within the lake basin since the retreat of the Wisconsinian glaciers (< ≊ 11 000 years ago), and (iii) distinct reproductive niches may promote divergence in north temperate freshwater fish faunas.     

Genetic and morphological differentiation in Tephritis bardanae (Diptera: Tephritidae): evidence for host-race formation

The fruit fly Tephritis bardanae infests flower heads of two burdock hosts, Arctium tomentosum and A. minus. Observations suggest host-associated mating and behavioural differences at oviposition indicating host-race status. Previously, flies from each host plant were found to differ slightly in allozyme allele frequencies, but these differences could as well be explained by geographical separation of host plants. In the present study, we explicitly test whether genetic and morphological variance among T. bardanae are explained best by host-plant association or by geographical location, and if this pattern is stable over a 10-year period. Populations of A. tomentosum flies differed significantly from those of A. minus flies in (i) allozyme allele frequencies at the loci Pep-A and Pgd, (ii) mtDNA haplotype frequencies and (iii) wing size. In contrast, geographical location had no significant influence on the variance estimates. While it remains uncertain whether morphometric differentiation reflects genotypic variability or phenotypic plasticity, allozyme and mtDNA differentiation is genetically determined. This provides strong evidence for host-race formation in T. bardanae. However, the levels of differentiation are relatively low indicating that the system is in an early stage of divergence. This might be due to a lack of time (i.e. the host shift occurred recently) or due to relatively high gene flow preventing much differentiation at loci not experiencing selection.     

TEMPORAL CHANGE OF MITOCHONDRIAL DNA HAPLOTYPE FREQUENCIES AND FEMALE EFFECTIVE SIZE IN A BROWN TROUT (SALMO TRUTTA) POPULATION

We report data on genetic drift of mitochondrial DNA (mtDNA) haplotypes in a natural brown trout (Salmo trutta) population in Sweden. Large temporal frequency shifts were observed over the 14 consecutive year classes studied. The observed rate of drift was used to estimate the effective size of the population. This effective size applies to the female segment of the population as mtDNA is maternally inherited. The magnitude of mtDNA haplotype frequency change is compared with the corresponding allele frequency changes at 14 allozyme loci in the same population. The female effective size is estimated as 58, which is approximately half the effective size of 97 for the total population (both sexes) previously obtained from the shifts of allozyme allele frequencies.     

Spatial heterogeneity of mitochondrial DNA and allozymes among populations of white-tailed deer and mule deer.

A white-tailed deer (Odocoileus virginianus) population in northeastern Minnesota and a mule deer (O. hemionus) population in the Bridger Mountains Montana, have previously been shown to be spatially subdivided into contiguous subpopulations. We assessed the degree of genetic differentiation among subpopulations and tested the hypothesis that differentiation will be greater for mitochondrial DNA (mtDNA) than for nuclear-encoded allozymes. Differentiation of the white-tailed deer subpopulations was significant for two allozyme loci but not for mtDNA, and the overall degree of differentiation was low. Gene flow, recent founding of the subpopulations, and polygamous breeding structure may all have contributed to this pattern. Greater differentiation was evident among disjunct populations than between the contiguous subpopulations of white-tailed deer. The contiguous mule deer subpopulations were significantly differentiated for mtDNA and one allozyme locus. Differentiation was greater for mtDNA than for allozymes. These results are consistent with demographic data that indicate mule deer males disperse more than do females. Disjunct mule deer populations may be similar or dramatically different in mtDNA haplotype frequencies that do not necessarily vary with geographic distance. Current and historical gene flow and breeding structure will influence population genetic patterns.     

DEVELOPMENT OF THE HYBRID SWARM BETWEEN PECOS PUPFISH (CYPRINODONTIDAE: CYPRINODON PECOSENSIS) AND SHEEPSHEAD MINNOW (CYPRINODON VARIEGATUS): A PERSPECTIVE FROM ALLOZYMES AND mtDNA

A comparison of allozyme and mtDNA frequencies was used for insight into a situation in the Pecos River, Texas where contact between the endemic pupfish (Cyprinodon pecosensis) and an introduced congener (C. variegatus) has resulted in rapid, geographically extensive genetic introgression. Temporal changes in mean frequencies of diagnostic allozyme markers indicate that the clinal pattern of introduced genetic material (Echelle and Connor 1989) is slowly decreasing in amplitude. Significant rank concordance in diagnostic allele frequencies among sites and across sampling years indicates directional influences upon temporal allele frequency change. These observations are consistent with the theory of gene flow in neutral clines. Levels of introgression indicated by each of four allozyme loci and mtDNA were roughly equivalent. The early history of the hybrid swarm is explained by genetic swamping, possibly mediated by selection for C. variegatus or C. variegatus × C. pecosensis, at a time when the normally abundant endemic species had been catastrophically depleted. High frequencies of an introduced GPI-A allele in all samples of intergrades suggests that the introduced genome originated with a single founding event.     

Divergent mitochondrial lineages arose within a large,panmictic population of the Savannah sparrow (Passerculus sandwichensis)

Unusual patterns of mtDNA diversity can reveal interesting aspects of a species’ biology. However, making such inferences requires discerning among the many alternative scenarios that could underlie any given mtDNA pattern. Next‐generation sequencing methods provide large, multilocus data sets with increased power to resolve unusual mtDNA patterns. A mtDNA‐based phylogeography of the Savannah sparrow (Passerculus sandwichensis) previously identified two sympatric, but divergent (~2%) clades within the nominate subspecies group and a third clade that consisted of birds sampled from northwest Mexico. We revisited the phylogeography of this species using a population genomic data set to resolve the processes leading to the evolution of sympatric and divergent mtDNA lineages. We identified two genetic clusters in the genomic data set corresponding to (a) the nominate subspecies group and (b) northwestern Mexico birds. Following divergence, the nominate clade maintained a large, stable population, indicating that divergent mitochondrial lineages arose within a panmictic population. Simulations based on parameter estimates from this model further confirmed that this demographic history could produce observed levels of mtDNA diversity. Patterns of divergent, sympatric mtDNA lineages are frequently interpreted as admixture of historically isolated lineages. Our analyses reject this interpretation for Savannah sparrows and underscore the need for genomic data sets to resolve the evolutionary mechanisms behind anomalous, locus‐specific patterns.     

AFLP genome scans suggest divergent selection on colour patterning in allopatric colour morphs of a cichlid fish

Genome scan-based tests for selection are directly applicable to natural populations to study the genetic and evolutionary mechanisms behind phenotypic differentiation. We conducted AFLP genome scans in three distinct geographic colour morphs of the cichlid fish Tropheus moorii to assess whether the extant, allopatric colour pattern differentiation can be explained by drift and to identify markers mapping to genomic regions possibly involved in colour patterning. The tested morphs occupy adjacent shore sections in southern Lake Tanganyika and are separated from each other by major habitat barriers. The genome scans revealed significant genetic structure between morphs, but a very low proportion of loci fixed for alternative AFLP alleles in different morphs. This high level of polymorphism within morphs suggested that colour pattern differentiation did not result exclusively from neutral processes. Outlier detection methods identified six loci with excess differentiation in the comparison between a bluish and a yellow-blotch morph and five different outlier loci in comparisons of each of these morphs with a red morph. As population expansions and the genetic structure of Tropheus make the outlier approach prone to false-positive signals of selection, we examined the correlation between outlier locus alleles and colour phenotypes in a genetic and phenotypic cline between two morphs. Distributions of allele frequencies at one outlier locus were indeed consistent with linkage to a colour locus. Despite the challenges posed by population structure and demography, our results encourage the cautious application of genome scans to studies of divergent selection in subdivided and recently expanded populations.     

Incongruent estimates of population differentiation among brook charr, Salvelinus fontinalis, from Cape Race, Newfoundland, Canada, based upon allozyme and mitochondrial DNA variation

We determined the amount and temporal stability of genetic differentiation among brook cham sampled from five rivers on Cape Race, Newfoundland, with an electrophoretic analysis of 42 protein coding loci. Fish from four of these rivers were analysed for restriction fragment length polymorphisms in mitochondrial DNA (mtDNA). A single mtDNA clone was observed in all rivers sampled, except one, where 47% offish were from a different and relatively divergent clone (0.31 % sequence divergence). In contrast, Cape Race brook charr show large amounts of genetic differentiation at six enzyme coding loci; Nei's genetic distance ranged between 0,020 and 0.048. This differentiation is relatively stable as no significant differences in allele frequencies were detected between fish sampled from two rivers over two consecutive years. The most divergent population based on protein polymorphism is not that with two mtDNA clonal lineages. In contrast to the commonly held view, mtDNA analyses do not necessarily provide greater resolution of population structure than allozyme analyses.     

Molecular and morphological divergence in the butterfly genus Lycaeides (Lepidoptera: Lycaenidae) in North America: evidence of recent speciation

Male genital morphology, allozyme allele frequencies and mtDNA sequence variation were surveyed in the butterfly species Lycaeides idas and L. melissa from across much of their range in North America. Despite clear differences in male genital morphology, wing colour patterns and habitat characteristics, genetic variation was not taxonomically or geographically structured and the species were not identifiable by either genetic data set. Genetic distances (Nei's D=0.002–0.078, calculated from allozyme data) between all populations of both species were within the range commonly observed for conspecific populations of other butterflies. The most frequent mtDNA haplotype was present in individuals of both species in populations from southern California to Wisconsin. We conclude that speciation has probably happened recently and the lack of genetic differentiation between the species is the product of either (1) recent or ongoing gene flow at neutral loci, and/or (2) an insufficiency of time for lineage sorting. The evolution of male genital morphology, wing colour patterns and ecological characteristics has proceeded more rapidly than allozyme or mtDNA evolution.     

Differential patterns of spatial divergence in microsatellite and allozyme alleles: further evidence for locus‐specific selection in the acorn barnacle,Semibalanus balanoides?

We compared patterns of genetic structure at potentially selected (two allozyme loci) and neutral molecular markers (six microsatellite loci) in the acorn barnacle, Semibalanus balanoides from the Gulf of St. Lawrence. Our results confirmed the presence of a geographical shift in alleles MPI and GPI near the Miramichi River. In contrast, no significant patterns of population differentiation among samples located north and south of the river mouth were detected for four of six microsatellite loci. However, analysis of molecular variance (amova) at individual loci revealed that a significant proportion of the total variance in allele frequencies was partitioned among samples located north and south of the river for both the allozyme and the other two microsatellite loci. The two most common alleles at these microsatellites showed frequencies that were highly correlated (r = 0.65-0.74, P < 0.05) with those of the MPI*2 allele, perhaps because of either physical linkage or epistasis. The two allozyme loci were significantly correlated in barnacles located north of the Miramichi River (r = 0.86, P < 0.05). Overall, our results supported the hypothesis that the broad scale pattern of allozyme allelic shifts is maintained by selection. They also indicated that microsatellites may not always behave in a neutral way and must be used cautiously, especially when evidence for genetic structuring relies on only a few assayed loci.     

Unequal allelic frequencies at the self‐incompatibility locus within local populations of Prunus avium L.: an effect of population structure?

In this paper, we investigated the genetic structure and distribution of allelic frequencies at the gametophytic self-incompatibility locus in three populations of Prunus avium L. In line with theoretical predictions under balancing selection, genetic structure at the self-incompatibility locus was almost three times lower than at seven unlinked microsatellites. Furthermore, we found that S-allele frequencies in wild cherry populations departed significantly from the expected isoplethic distribution towards which balancing selection is expected to drive allelic frequencies (i.e. identical frequency equal to the inverse of the number of alleles in the population). To assess whether this departure could be caused either by drift alone or by population structure, we used numerical simulations to compare our observations with allelic frequency distributions expected : (1) within a single deme from a subdivided population with various levels of differentiation; and (2) within a finite panmictic population with identical allelic diversity. We also investigated the effects of sample size and degree of population structure on tests of departure from isoplethic equilibrium. Overall, our results showed that the observed allele frequency distributions were consistent with a model of subdivided population with demes linked by moderate migration rate.     

Genetic structure and dispersal of Macrobrachium australiense (Decapoda: Palaemonidae) in western Queensland, Australia

SUMMARY 1. The freshwater prawn, Macrobrachium australiense (Decapoda: Palaemonidae), is an abundant species throughout the rivers of western Queensland, and it is thought to have effective dispersal capabilities. Given the very low topographical relief of the western Queensland landscape and the vast nature of episodic flooding, it was predicted that widespread dispersal in this species would occur within, and possibly between the four major catchments of the region: the Darling, Bulloo, Cooper and Diamantina. We analysed eight polymorphic allozyme systems and a fragment of the cytochrome c oxidase subunit 1 (COI) mitochondrial DNA (mtDNA) gene to determine the extent of recent and historical patterns of dispersal at nested spatial scales, within and between catchments.
2. Large and significant levels of allozyme and mtDNA differentiation were revealed among all catchments, indicating that dispersal of M. australiense does not occur across catchment boundaries, although this species is reportedly capable of overland movement. In contrast, no significant patterns of genetic differentiation were resolved between major subcatchments of the Darling and Cooper, or between sites within these subcatchments, indicating that populations of M. australiense are panmictic within catchments.
3. The MtDNA data resolved two divergent and reciprocally monophyletic clades, with the first representing the Darling catchment, and the second corresponding to the Bulloo, Cooper and Diamantina catchments. We postulate that extreme variation in historical climatic patterns and palaeohydrologic conditions played an important role in shaping the population structure of M. australiense throughout western Queensland during the Quaternary.     

SELECTION ON THE COLOR POLYMORPHISM IN HAWAIIAN HAPPY-FACE SPIDERS: EVIDENCE FROM GENETIC STRUCTURE AND TEMPORAL FLUCTUATIONS

Throughout this century genetic polymorphisms for color have been widely used as a research tool to allow insights into key evolutionary processes. Although color variants can often be diverse within populations, frequencies of different morphs may be similar across populations, either as a result of balancing selection or gene flow. Under these circumstances selection can be extremely difficult to demonstrate. Here we test for balancing selection on the naturally occurring color forms of the Hawaiian happy-face spider, Theridion grallator with two approaches. First, allozyme loci are used to generate a null model against which to test selection. Frequencies of alleles involved in the color polymorphism of T. grallator are used to generate another estimate for comparison. The results suggest that statistically similar frequencies of color morphs among populations of T. grallator may be maintained by some form of balancing selection. Second, we make use of an unusual event in which the normally stable frequencies of unpatterned and patterned morphs within a population were found to have shifted toward an excess of unpatterned morphs. We scored offspring of all fertilized, unpatterned (bottom-recessive) females found during this period of skewed morph frequencies and also in a year when morph frequencies were normal to deduce paternal color phenotypes. Mating was found to be random in the normal year, but in the perturbed year females had mated with rare (patterned) males twice as frequently as expected on the basis of the frequency of this morph type in the population. Both of these results are consistent with selection operating on the color polymorphism, and we speculate that apostatic selection, perhaps mediated by bird predators, may provide the mechanism.     

GEOGRAPHIC DISTRIBUTION OF MOLECULAR VARIANCE WITHIN THE BLUE MARLIN (MAKAIRA NIGRICANS): A HIERARCHICAL ANALYSIS OF ALLOZYME,SINGLE-COPY NUCLEAR DNA,AND MITOCHONDRIAL DNA MARKERS

This study presents a comparative hierarchical analysis of variance applied to three classes of molecular markers within the blue marlin (Makaira nigricans). Results are reported from analyses of four polymorphic allozyme loci, four polymorphic anonymously chosen single-copy nuclear DNA (scnDNA) loci, and previously reported restriction fragment length polymorphisms (RFLPs) of mitochondrial DNA (mtDNA). Samples were collected within and among the Atlantic and Pacific Oceans over a period of several years. Although moderate levels of genetic variation were detected at both polymorphic allozyme (H = 0.30) and scnDNA loci (H = 0.37), mtDNA markers were much more diverse (h = 0.85). Allele frequencies were significantly different between Atlantic and Pacific Ocean samples at three of four allozyme loci and three of four scnDNA loci. Estimates of allozyme genetic differentiation (θ_O) ranged from 0.00 to 0.15, with a mean of 0.08. The θ_O values for scnDNA loci were similar to those of allozymes, ranging from 0.00 to 0.12 with a mean of 0.09. MtDNA RFLP divergence between oceans (θ_O = 0.39) was significantly greater than divergence detected at nuclear loci (95% nuclear confidence interval = 0.04–0.11). The fourfold smaller effective population size of mtDNA and male-mediated gene flow may account for the difference observed between nuclear and mitochondrial divergence estimates.     

Analyses of Nuclear ldhA Gene and mtDNA Control Region Sequences of Atlantic Northern Bluefin Tuna Populations

There has been considerable debate about whether the Atlantic northern bluefin tuna exist as a single panmictic unit. We have addressed this issue by examining both mitochondrial DNA control region nucleotide sequences and nuclear gene ldhA allele frequencies in replicate size or year class samples of northern bluefin tuna from the Mediterranean Sea and the northwestern Atlantic Ocean. Pairwise comparisons of multiple year class samples from the 2 regions provided no evidence for population subdivision. Similarly, analyses of molecular variance of both mitochondrial and ldhA data revealed no significant differences among or between samples from the 2 regions. These results demonstrate the importance of analyzing multiple year classes and large sample sizes to obtain accurate estimates when using allele frequencies to characterize a population. It is important to note that the absence of genetic evidence for population substructure does not unilaterally constitute evidence of a single panmictic population, as genetic differentiation can be prevented by large population sizes and by migration.     

Evaluating Genetic Diversity for the Conservation of the Threatened Galapagos Endemic Calandrinia galapagosa (Portulacaceae)

Calandrinia galapagosa (Portulacaceae) is a threatened plant endemic to San Cristobal Island, Galapagos. The species is threatened by goat herbivory: less than 1300 individuals remain in nine populations, three of which have been fenced for protection. It is not known whether these three populations are genetically representative of the species at large or whether the three distinct color morphs exhibited by the species represent different taxonomic units. We measured the frequency of 113 polymorphic amplified fragment length polymorphism markers in 189 specimens representing three different phenotypes from all nine populations to relate the genetic structure of the species to both its geological history and morphological variation. The analysis revealed high diversity within populations and low, but significant, differentiation between populations. In addition, no genetic diversification was observed across color morphs. We conclude that (1) regular gene flow has historically occurred between populations, and (2) individual color morphs are not separate taxa. We identify another population that should be fenced to ensure maximal protection of genetic variation. Our research serves as a case study of using population molecular genetics for optimizing protection strategies for rare plants in tropical archipelagoes affected by introduced herbivores.     

Cryptic species within the freshwater isopod Mesamphisopus capensis (Phreatoicidea: Amphisopodidae) in the Western Cape, South Africa: allozyme and 12S rRNA sequence data and morphometric evidence

The freshwater phreatoicidean isopod Mesamphisopus capensis has been regarded as the most widespread of the four Mesamphisopus species occurring in the Western Cape, South Africa. To determine whether this species was monotypic across its distribution over two mountainous regions, separated by a low-lying coastal plain remnant, genetic differentiation among populations from 11 localities was studied through allozyme electrophoresis of 12 loci and sequencing of a 338-bp 12S rRNA mtDNA fragment from representative individuals. Populations of the two regions were separated by a mean identity value of 0.477. Fixed allele differences at two loci distinguished these regions. Estimates of θ indicated substantial differentiation among populations across the entire sample, as well as within each of the regions. Topologies derived through parsimony and neighbour joining supported the monophyly of the two regions. On the basis of these topologies, allele frequencies and an allozyme dendrogram, five groups were identified. Discriminant function analyses, performed on body and pereopod variables independently, revealed these groups to be well differentiated with a high rate of correct a posteriori reclassification. Using genetic distance criteria these five distinct forms may be considered to be putative species. From a conservation perspective, the two regions can be seen to represent two evolutionarily significant units, while the five groups should be regarded as management units.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 235–253.     

Genetic differentiation among North Atlantic killer whale populations

Population genetic structure of North Atlantic killer whale samples was resolved from differences in allele frequencies of 17 microsatellite loci, mtDNA control region haplotype frequencies and for a subset of samples, using complete mitogenome sequences. Three significantly differentiated populations were identified. Differentiation based on microsatellite allele frequencies was greater between the two allopatric populations than between the two pairs of partially sympatric populations. Spatial clustering of individuals within each of these populations overlaps with the distribution of particular prey resources: herring, mackerel and tuna, which each population has been seen predating. Phylogenetic analyses using complete mitogenomes suggested two populations could have resulted from single founding events and subsequent matrilineal expansion. The third population, which was sampled at lower latitudes and lower density, consisted of maternal lineages from three highly divergent clades. Pairwise population differentiation was greater for estimates based on mtDNA control region haplotype frequencies than for estimates based on microsatellite allele frequencies, and there were no mitogenome haplotypes shared among populations. This suggests low or no female migration and that gene flow was primarily male mediated when populations spatially and temporally overlap. These results demonstrate that genetic differentiation can arise through resource specialization in the absence of physical barriers to gene flow.     

Systematics and phylogeography of the Taiwanese endemic minnow Candidia barbatus (Pisces: Cyprinidae) based on DNA sequence,allozymic, and morphological analyses

The phylogeographical patterns of the endemic minnow, Candidia barbatus, were examined using mitochondrial (mt) cytochrome b sequences, allozyme electrophoresis, and morphology. The 23 mtDNA haplotypes recovered were grouped into six lineages (A–F). According to a molecular clock of 1.7% per million years, we estimated that the colonization of C. barbatus to Taiwan took place three million years ago with several subsequent vicariance events. In the allozyme analysis, the Bayesian approach partitioned different populations into three clusters, with clusters I, II, and III mainly found in northern, central, and southern Taiwan, respectively. Morphologically, fish from north and south of the Tropic of Cancer had distinct numbers of lateral line scales and were separated into two morphs. The cohabitation of different mtDNA lineages with high allozymic and morphological similarities suggests that interbreeding amongst them is possible. Therefore, in spite of highly differentiated mtDNA, samples from different collecting sites should be considered a single species. Based on the current population distribution and levels of divergence, we propose that both orogenic activities and marine regression during glacial epoch were the two main forces shaping population differentiation of this species. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 613–632.