Genetic cohesion of Eresus walckenaeri (Araneae, Eresidae) in the eastern Mediterranean

The eresid spider genus Eresus is morphologically and ecologically conservative. At least three species occur in Europe. However, deep genetic divergence among geographical samples within two species, E. cinnaberinus and E. sandaliatus , may suggest more cryptic species. In the present study we investigate the genetic cohesion of the third species, Eresus walckenaeri , throughout its eastern Mediterranean distribution range, relative to the E. cinnaberinus–E. sandaliatus species complex. Eresus walckenaeri specimens were monophyletic. Genetic discreteness of E. walckenaeri in a region of sympatry with its sister species in Greece provides evidence for species integrity of E. walckenaeri within the European Eresus species complex. Eresus walckenaeri exhibited high concordance between geographical location and mtDNA genealogy. Two major phylogeographical clades were found in the Greek–Turkish and Syrian–Israel parts of the investigated area, respectively (∼6.5% sequence divergence). Concordance between geography and genetic divergence was further observed between Aegean island samples and their corresponding Greek and Turkish mainland samples, suggesting regional subdivision with gradual but potentially high dispersal propensity. Monophyly and limited regional distribution indicate Mediterranean endemic origin.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 86 , 1–9.     

Fine‐scale genetic structure overrides macro‐scale structure in a marine snail: nonrandom recruitment,demographic events or selection?

The planktotrophic littorinid species Littoraria flava occupies a continuous habitat on rocky shores close to brackish and freshwater sources. Previous studies of this species have shown a moderate genetic structure over a broad geographical scale, with high deviations from Hardy–Weinberg expectations in many allozymic loci. Local-scale subdivision in marine species with a long dispersal phase is unexpected, but occasionally found. Using a horizontal transect at three locations, we examined whether microscale and short-term subdivision also occurred in L. flava populations and, if so, whether this could explain the Hardy–Weinberg deviations. Littoraria flava showed even more structuring on a microgeographical scale (4–300 m) than on a large-scale (> 200 km). The Ewens–Watterson neutrality test showed that 18% of the tests deviated significantly from the neutrality model. A homogeneity test for each locus across samples within transects showed homogeneous and high F _IS values in many loci. These results and the apparent genetic patchiness within transects suggest that asynchronous spawning associated with recurrent colonizations in L. flava can explain the local differentiation without a recognizable pattern. In addition, there could be a balance between these factors and diversifying selection acting on different loci at different times and localities. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 23–36.     

Population genetic structure of New Zealand's endemic corophiid amphipods: evidence for allopatric speciation

Allozyme electrophoresis was used to examine population genetic structure at inter- and intraspecific levels for the New Zealand endemic corophiid amphipods, Paracorophium lucasi and P. excavatum. Individuals were collected from estuarine and freshwater habitats from North, South and Chatham Islands. Analyses of genetic structure among interspecific populations indicated clear allelic differentiation between the two Paracorophium species (Nei's genetic distance, D  = 1.62), as well as considerable intraspecific substructuring ( D  = 0.15–0.65). These levels of divergence are similar to interspecific levels for other amphipods and it is proposed that at least two groups from the P. lucasi complex and three from the P. excavatum complex correspond to sibling species. In most cases allopatry can account for the differentiation among the putative sibling species. For populations that share a common coastline we found low levels of differentiation and little or no correlation with geographical distance, suggesting that gene flow is adequate to maintain homogeneous population genetic structure. By contrast, populations on separate coastlines (i.e. isolated by land) showed moderate levels of geographical differentiation indicating restricted gene flow. The juxtaposition of population genetic and biogeographical data for Paracorophium in conjunction with the geological record infers past histories of glacial extirpation, and possible isolating effects of sea-level and landmass changes that have occurred throughout the Plio-Pleistocene.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 119–133.     

Genetic divergence of peripherally disjunct populations of the gastropod Batillariella estuarina in the Houtman Abrolhos Islands, Western Australia

Geographically disjunct populations are unusual in marine species, but the Houtman Abrolhos Islands, Western Australia, provide opportunities to study highly disjunct peripheral isolates of several species. The intertidal snail Batillariella estuarina occurs in isolated tidal ponds in the Abrolhos Islands, where it is at its northern limit, disjunct from mainland populations by 600-900 km. The species is thus disjunct both geographically and among the peripherally isolated populations in the Abrolhos Islands. Comparisons of allozymes at 11 polymorphic loci were made among populations from 10 ponds in the Abrolhos Islands and six sites from relatively continuous tidal flats at Albany, 900 km away, the nearest major set of populations. Among all 16 populations, subdivision was high (FST = 0.455). Although there were subtle differences between the geographical regions, the large majority of divergence occurred among the isolated ponds in the Abrolhos (FST = 0.441), and divergence on the tidal flats at Albany was only moderate (FST = 0.085). Characteristic of peripheral isolates, the pond populations have less polymorphism and fewer alleles than the more connected populations at Albany. Combined with evidence of genetic divergence in the gastropods Bembicium vittatum and Austrocochlea constricta, which have very similar geographical distributions to that of B. estuarina, these results indicate the potential evolutionary significance of peripherally isolated marine populations in the unusual habitats of the Abrolhos Islands.     

Phylogeography of the freshwater crayfish Cherax destructor Clark (Parastacidae) in inland Australia: historical fragmentation and recent range expansion

The yabby, Cherax destructor Clark, is the most widespread species in the most widespread genus of Australian freshwater crayfish. It has a distribution that spans several distinct drainage basins and biogeographical regions within semiarid and arid inland Australia. Here we report a study designed to investigate patterns of genetic variation within the species and hypotheses put forward to account for its extensive distribution using DNA sequences from the mitochondrial 16S rRNA gene region. Results of phylogenetic analyses contradicted previous allozyme data and revealed relatively deep phylogenetic structure in the form of three geographically correlated clades. The degree of genetic divergences between clades (8–15 bp) contrasted with the relatively limited haplotype diversity within clades (1–3 bp). Network-based analyses confirmed these results and revealed genetic structure on both larger and more restricted geographical scales. Nevertheless some haplotypes and 1-step clades had large distributions, some of which crossed boundaries between river basins and aquatic biogeographical regions. Thus both older and more recent historical processes, including fragmentation on a larger geographical scale and more recent range expansion on a local scale, appear to be responsible for the observed pattern of genetic variation within C. destructor . These results support elements of alternative hypotheses previously put forward to account for the evolutionary history of C. destructor and the origin of its large distribution.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 539–550.     

High genetic variability in Neotropical myophilous orchids

The patterns of genetic variability of seven Bulbophyllum Thouars species were investigated using 14 enzymatic loci, and compared with those of other Neotropical myophilous orchid species. The genetic variability estimated was very high ( H _e = 0.39–0.61, P  = 86–100%, A  = 2.6–3.8), despite the small population sizes, which may be explained by the vegetative reproduction. Of the three species with multiple populations, only B. epiphytum showed moderate values of genetic structuring. Environmental characteristics and seed dispersal mechanisms are important in understanding the differences in the population structure observed. Almost all pairs of species showed low genetic similarity, indicating a long period of divergence. Our results suggest that B. adiamantinum and B. insectiferum , species currently classified in different sections of the genus, are more closely related than previously thought.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 33–40.     

Development, phylogeny, and taxonomy of Bostrycapulus (Caenogastropoda: Calyptraeidae), an ancient cryptic radiation

Calyptraeid gastropods are well know for the taxonomic difficulties caused by their simple, phenotypically variable shells. In this paper I demonstrate that what was previously considered to be a single species, Crepidula aculeata , is an ancient (3–15 Myr) cryptic species complex made up of at least eight species, and that this group should be placed in the genus Bostrycapulus . Despite the difficulty in finding diagnostic adult shell and anatomical features upon which species can be unambiguously identified, DNA sequences, protoconch morphology, embryonic morphology and developmental characters clearly differentiate these eight species. A single species with direct development and nurse eggs is present in the South Atlantic, and a species with planktotrophic development occurs in the equatorial Pacific. The species from Japan, Australia, Florida, the Pacific coasts of Mexico and Central America, and the Cape Verde Islands all have direct development. Most of these species are separated by > 15% divergence in COI sequence data. The fossil record of Bostrycapulus goes back to the Miocene, which agrees with genetic estimates of divergences within the genus ranging from 3 to 15 Mya. Surprisingly, these ancient species differ only slightly in morphology from each other and genetic differentiation does not correlate with geographical distance. I revise the genus Bostrycapulus on the basis of differences in adult morphology, embryonic morphology, mode of development, protoconch morphology, and DNA sequence data. I also describe four new species ( B. pritzkeri sp. nov., B. odites sp. nov., B. latebrus sp. nov. and B. urraca sp. nov. ) and remove three others ( B. gravispinosus , B. calyptraeformis , and B.  cf. tegulicius ) from synonymy with B. aculeatus .  © 2005 The Linnean Society of London, Zoological Journal of the Linnean Society , 2005, 144 , 75−101.     

Genetic differentiation on multiple spatial scales in an ecotype-forming marine snail with limited dispersal: Littorina saxatilis

The population genetic structure of marine species lacking free-swimming larvae is expected to be strongly affected by random genetic drift among populations, resulting in genetic isolation by geographical distance. At the same time, ecological separation over microhabitats followed by direct selection on those parts of the genome that affect adaptation might also be strong. Here, we address the question of how the relative importance of stochastic vs. selective structuring forces varies at different geographical scales. We use microsatellite DNA and allozyme data from samples of the marine rocky shore snail Littorina saxatilis over distance scales ranging from metres to 1000 km, and we show that genetic drift is the most important structuring evolutionary force at distances > 1 km. On smaller geographical scales (< 1 km), divergent selection between contrasting habitats affects population genetic structure by impeding gene flow over microhabitat borders (microsatellite structure), or by directly favouring specific alleles of selected loci (allozyme structure). The results suggest that evolutionary drivers of population genetic structure cannot a priori be assumed to be equally important at different geographical scales. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 31–40.     

The molecular systematics of Leiopotherapon unicolor (Günther, 1859): testing for cryptic speciation in Australia's most widespread freshwater fish

Leiopotherapon unicolor is the most widespread freshwater fish species in Australia. A comprehensive allozyme and mitochondrial DNA 16S rRNA data set was assembled from 141 specimens of L. unicolor collected Australia-wide in order to test for cryptic speciation in this far-ranging species. Surprisingly, little genetic diversity was observed within L. unicolor and provided no evidence for the existence of cryptic species within this lineage. In contrast, a small sample set of L. aheneus used as the outgroup showed two highly divergent haplotypes strongly suggestive of cryptic speciation. L. unicolor has a number of ecological and life history attributes that may explain the lack of significant genetic divergence over substantial geographical distances. The occurrence of other widespread fish and crustacean species that also display only limited genetic diversity indicate that climate conditions more favourable to dispersal across central and northern Australia than is suggested by the extent of present-day aridity have occurred in the relatively recent geological past.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 537–552.     

Crosses between frog populations reveal genetic divergence in larval life history at short geographical distance

A number of studies have documented interpopulation divergence in amphibian larval life-history traits across latitudes. Because many frogs are philopatric and have a patchy habitat distribution, genetic divergence could also exist on a much smaller geographical scale, revealed by recent estimates of population divergence using molecular markers. Whether this divergence is reflected in phenotypic traits is virtually unknown. Using artificial fertilization, individuals of the common frog, Rana temporaria , were crossed from two populations situated 130 km apart and differing in population size. The pattern of size at metamorphosis showed evidence of non-additive effects, as demonstrated by a significant interaction between male and female population of origin. Outbreeding resulted in an increase in metamorph size when eggs from the small population were fertilized with sperm from the large population. In the reciprocal cross, however, the pattern was in the opposite direction, with no significant effect of male population of origin. Genetic divergence of populations separated by a relatively short geographical distance may be more common in frogs than previously acknowledged, with potential implications for conservation of declining amphibian species.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 189–195.     

Genetic and morphological divergence reveals local subdivision of perch (Perca fluviatilis L.)

The level of gene flow is an important factor influencing genetic differentiation between populations. Typically, geographic distance is considered to be the major factor limiting dispersal and should thus only influence the degree of genetic divergence at larger spatial scales. However, recent studies have revealed the possibility for small-scale genetic differentiation, suggesting that the spatial scale considered is pivotal for finding patterns of isolation by distance. To address this question, genetic and morphological differentiation were studied at two spatial scales (range 2–13 km and range 300 m to 2 km) in the perch ( Perca fluviatilis L.) from the east coast archipelago of Sweden, using seven microsatellite loci and geometric morphometrics. We found highly significant genetic differentiation between sampled locations at both scales. At the larger spatial scale, the distance per se was not affecting the level of divergence. At the small scale, however, we found subtle patterns of isolation by distance. In addition, we also found morphological divergence between locations, congruent with a spatial separation at a microgeographic scale, most likely due to phenotypic plasticity. The present study highlights the importance of geographical scale and indicates that there might be a disparity between the dispersal capacity of a species and the actual movement of genes. Thus, how we view the environment and possible barriers to dispersal might have great implications for our ability to fully understand the evolution of genetic differentiation, local adaptation, and, in the end, speciation.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 746–758.     

Genetic variation and structure in native populations of the fire ant Solenopsis invicta: evolutionary and demographic implications

We studied population genetic variation and structure in the fire ant Solenopsis invicta using nuclear genotypic and mitochondrial DNA (mtDNA) sequence data obtained from samples collected throughout its native range. Geographic populations are strongly differentiated at both genomes, with such structure more pronounced in Brazil than in Argentina. Higher-level regional structure is evident from the occurrence of isolation-by-distance patterns among populations, the recognition of clusters of genetically similar, geographically adjacent populations by ordination analysis, and the detection of an mtDNA discontinuity between Argentina and Brazil coinciding with a previously identified landform of biogeographical relevance. Multiple lines of evidence from both genomes suggest that the ancestors of the ants we studied resembled extant northern Argentine S. invicta , and that existing Brazilian populations were established more recently by serial long-distance colonizations and/or range expansions. The most compelling evidence for this is the corresponding increase in F _K (a measure of divergence from a hypothetical ancestor) and decrease in genetic diversity with distance from the Corrientes population in northern Argentina. Relatively deep sequence divergence among several mtDNA clades, coupled with geographical partitioning of many of them, suggests prolonged occupation of South America by S. invicta in more-or-less isolated regional populations. Such populations appear, in some cases, to have come into secondary contact without regaining the capacity to freely interbreed. We conclude that nominal S. invicta in its native range comprises multiple entities that are sufficiently genetically isolated and diverged to have embarked on independent evolutionary paths.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 541–560.     

Relationships in Patagonian species of Berberis (Berberidaceae) based on the characterization of rDNA internal transcribed spacer sequences

Sequence analysis of the internal transcribed spacer (ITS) of the 18S(ITS1)-5.8S-26S(ITS2) rDNA region was performed in order to analyse the phylogenetic relationships between 13 Patagonian species of the genus Berberis (Berberidaceae). The divergence values between the pairwise sequence in the studied Patagonian species were in the range 2.9–22.9%. The lengths of the ITS1 and ITS2 sequences were in the range 227–231 bp and 220–224 bp, respectively, and the 5.8S sequence was 159 bp throughout all species . B. microphylla sensu Landrum does not appear to be monophyletic based on current sampling. Indeed, we suggest that B. microphylla should be distinguished from B. buxifolia , B. parodii , and B. heterophylla . ITS sequences, together with data obtained from morphological, biochemical, amplified fragment length polymorphism, and cytological characterizations, support the existence of diploid and polyploid hybrid speciation in the genus.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 321–328.     

Population genetics, phylogeography and speciation of Cystodytes (Ascidiacea) in the western Mediterranean Sea

Several morphotypes that so far have been attributed to the allegedly cosmopolitan ascidian Cystodytes dellechiajei occur in the Mediterranean Sea. Colour variation is the difference most frequently reported. In this study, we addressed the genetic structure of this ascidian in relation to geographical location and colour morph. Partial sequences of the gene cytochrome  c oxidase subunit 1 (COI) were obtained from seven populations of the western Mediterranean, encompassing eight colour varieties. All population genetic analyses (exact test, pairwise F _ST, hierarchical analysis of molecular variance, multidimensional scaling, nested clade analysis) indicated clearly that differences between colour morphs are large enough to obscure any geographical differentiation when colours are combined within localities. When variance due to colour divergence was removed, however, a significant geographical variability between localities remained. The genetic divergence between the colour morphs analysed was significant in comparisons of the brown and purple forms with the others, but not among the green, blue, and white morphs. Phylogeographic analyses suggest that population fragmentation and range expansions have shaped the present-day distribution of the haplotypes. Taken together with existing chemotype information, our results indicate that several species are present in the area, and that a thorough revision of the genus is necessary.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 203–214.     

Phylogeography of the Iberian populations of Mioscirtus wagneri (Orthoptera: Acrididae), a specialized grasshopper inhabiting highly fragmented hypersaline environments

Phylogeographic studies from western Palaearctic have generally focused on species able to disperse and track their emerging suitable habitats after the last ice age. However, data on species whose biogeographical histories differ from this bulk of Palaearctic fauna are scarce. This is clearly the case of some specialized organisms inhabiting inland hypersaline environments, which are likely to have had a wider distribution range during the late Tertiary and may have persisted through the Pleistocene to the present day only constituting relict populations. In this study, we use partial sequences from two mitochondrial genes [16S rRNA (16S) and cytochrome oxidase subunit II (COII)] to investigate the phylogeography of the Iberian populations of Mioscirtus wagneri (Orthoptera: Acrididae), a specialized grasshopper exclusively inhabiting hypersaline low grounds. Our results show that M. wagneri exhibits a marked phylogeographical structure, forming three main clades which correspond with populations located in north-east, central–south-east and south-west Iberia. These geographical areas did not share any haplotype, indicating that gene flow between them is absent. Nested clade analyses revealed that these lineages have probably evolved in allopatry and data on sequence divergence suggest population fragmentation from the Early Pleistocene. Overall, these results provide a broader perspective on the contribution of historical climate/geological events to biogeographical patterns of organisms currently forming relict populations of great conservation concern.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 623–633.     

Genetic structure and differentiation of Gentiana atuntsiensis W. W. Smith and G. striolata T. N. Ho (Gentianaceae) as revealed by ISSR markers

The Hengduan Mountains are the core region of the Himalaya hotspot, and are renowned for their high levels of endemism. Gentiana atuntsiensis and G. striolata are two closely related and morphologically similar species endemic to this region. In this study, inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure within these two species, as well as the differentiation between them. An analysis of molecular variance-derived estimate demonstrated only 13.5% of genetic differentiation between the two species. Considering their adjacent distribution patterns, low genetic divergence, and clear clustering into two groups, it is hypothesized that the two species arose from a rapid and recent speciation event induced mainly by geographical isolation. A relatively high level of genetic diversity was revealed in each species (Shannon's index of diversity: H _sp = 0.324 and 0.391; H _pop = 0.225 and 0.274; for G. atuntsiensis and G. striolata , respectively). Most of the genetic variation was partitioned within populations ( Ø _ST = 0.232 and 0.226 in G. atuntsiensis and G. striolata , respectively). The large population sizes, outcrossing breeding system, and small, light seeds that disperse widely may explain the genetic structure in both species.  © 2007 The Linnean Society of London. Botanical Journal of the Linnean Society , 2007, 154 , 225–232.     

Microsatellite DNA polymorphism confirms reproductive isolation and reveals differences in population genetic structure of cryptic pipistrelle bat species

Previous studies have indicated that the common European pipistrelle bat ( Pipistrellus pipistrellus ) comprises two cryptic species, P. pipistrellus and Pipistrellus pygmaeus , which differ in echolocation call frequency and mitochondrial DNA sequence. However, levels of divergence based on nuclear markers have not been examined, and hence the potential for male-mediated gene flow between the species cannot be discounted. Moreover, little is known about population structure and migration patterns in either species. Here, we describe the use of microsatellites to investigate nuclear DNA differentiation between, and the pattern of population genetic structure within, the two cryptic pipistrelle species. In total, 1300 individuals from 82 maternity colonies were sampled across the British Isles and Continental Europe. We show, using multivariate analyses, that colonies of the same species are generally genetically more similar to each other than to those from the other species regardless of geographical location. Our findings support the hypothesis that the species are reproductively isolated. Significant patterns of genetic isolation by distance were identified in both species, indicating that mating may occur before any long-distance autumnal migration. The presence of a sea channel does not confer higher levels of genetic differentiation among colonies over and above distance alone in either species. Differences in genetic population structure were identified between the species, with P. pipistrellus showing a wider range of levels of genetic differentiation among colonies and a stronger relationship between genetic and geographical distance than P. pygmaeus . Differences in dispersal, mating behaviour, colony size and/or postglacial colonization patterns could contribute to the differences observed.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 539–550.     

Population genetics,gene flow,and biogeographical boundaries of Carcinus aestuarii (Crustacea: Brachyura: Carcinidae) along the European Mediterranean coast

Carcinus aestuarii Nardo, 1847 is a widespread coastal crab species throughout the Mediterranean Sea with a pelagic larval phase. This species tolerates a wide range of environmental conditions and typically inhabits fragmented habitats, such as embayments, lagoons and estuaries. It is therefore a good candidate species for studying and testing different phylogeographical hypotheses in the Mediterranean Sea. By contrast to its Atlantic sister species, Carcinus maenas, studies on the population genetic structure of C. aestuarii in its native range are still scarce. In the present study, specimens from along the European Mediterranean Sea were collected and DNA‐sequenced and analyses were applied to discriminate between present day and historical factors influencing the population genetic structure of this species. The results obtained demonstrate the existence of two genetically distinct geographical groups, corresponding to the eastern and western Mediterranean, with further subdivision within the East Mediterranean Basin. A strong asymmetric gene flow was recorded toward the Eastern Basin, which may play a crucial role in shaping the present day biogeographical patterns of this species and potentially other sympatric ones with pelagic larvae. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 771–790.     

Delimiting species in the hoplonemertean genus Tetrastemma (phylum Nemertea): morphology is not concordant with phylogeny as evidenced from mtDNA sequences

The marine genus Tetrastemma contains monostiliferous hoplonemertean (phylum Nemertea) species which mostly undergo direct development with no free-swimming stages of larvae. The lack of a pelagic phase, and the fact that many benthic species lay eggs attached to the bottom substrate, are obvious restrictions on dispersal and gene flow. Nevertheless, some of the species, for example T. candidum and T. melanocephalum , are described as ubiquitous and are reported from waters all over the world. We studied genetic variation and evolutionary relationships in order to assess whether they are concordant with external characters in samples of nine morphologically distinct forms formally named as different Tetrastemma species, from different geographical localities. We estimated the phylogeny and species network based on 539 base pairs of the mitochondrial protein-coding gene cytochrome oxidase-1 (CO1) for 30 ingroup specimens. From this, we assessed the evolutionary history and phylogenetic relationships between these forms. We conclude that in most cases there was no correspondence between evolutionary lineage and morphotype. Our results thus indicate that morphological species delimitation in nemerteans may be questionable, and that this in turn may have a profound effect upon estimates of species diversity within the phylum.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 86 , 201–212.