The Straits of Gibraltar: barrier or bridge to Ibero‐Moroccan bat diversity?

Genetic divergence in bat communities was assessed on both sides of the Straits of Gibraltar and cryptic diversity was examined. Screening was carried out using partial sequences of the mitochondrial (mt)DNA cytochrome b gene on 399 individual bats belonging to the 18 species found on both sides of the Straits of Gibraltar. For those bats that showed important genetic discontinuities, molecular markers (ND1 and nuclear RAG2 genes) were added to expand the sampling process. Phylogenetic reconstructions were obtained using maximum parsinomy, genetic distances, maximum likelihood, and Bayesian criteria. As an estimate of bats' flight performance, we measured for each species the wing aspect ratio and wing loading indexes, and correlated them with the maximum pairwise genetic distances obtained between southern Iberian and northern Moroccan populations. Genetic mtDNA distances between populations on both continents exceed 5% in seven out of 18 bat species analysed and unknown lineages were uncovered within the species complexes Myotis nattereri and Myotis mystacinus . We did not find a general pattern in the degree of permeability of the Straits of Gibraltar for bats. Genetic distances were not correlated with the ability to cross the Straits. Our study shows that the cryptic diversity uncovered among bats continues to increase as more regions are studied.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 434–450.     

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.     

An evaluation of the taxonomic distinctness of two Geonoma macrostachys (Arecaceae) varieties based on intersimple sequence repeat (ISSR) variation

The latest taxonomic account for the palms of the Amazon recognizes two common and widespread varieties of Geonoma macrostachys (Arecaceae) in western Amazonia: acaulis and macrostachys . These varieties are joined by intermediates, which obscure their taxonomic boundaries. An evaluation of the genetic distinctness between G. macrostachys varieties at a local and regional scale is presented. Thirty-one G. macrostachys individuals were randomly sampled from four Peruvian tropical moist forests. Twenty-seven intersimple sequence repeat (ISSR) primers were used, and all clear and reproducible bands were scored and analysed. Of the 99 ISSR bands produced, 51.52% were polymorphic. Nei's gene diversity value ( H ) was 0.2274 and Shannon's information index ( I ) was 0.3237. Clustering, ordination, and analyses of molecular variance (AMOVAs) suggested a lack of genetic distinctness between varieties at the regional level. Individuals clustered by geographical provenance, and two main groups were identified. A significant genetic differentiation between sympatric varieties occurred only in the northern locality (Φ_ST = 0.359, P  = 0.000). A hierarchical AMOVA revealed that the genetic diversity in G. macrostachys mainly lies between localities (76.58%). Mantel's test revealed no significant correlation between the geographical and genetic distances between individuals. We do not support the recognition of the two taxonomic varieties studied.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 381–392.     

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.     

Parmelina quercina (Parmeliaceae, Lecanorales) includes four phylogenetically supported morphospecies

Morphological and phylogenetic relationships of the worldwide Mediterranean lichen forming fungus, Parmelina quercina , have been studied. Specimens from western Europe, western North America and southern Australia were analysed using molecular data (nuITS rDNA, nuLSU rDNA and mtSSU rDNA) and selected morphological features (upper cortex maculae, scanning electron microscopy examination of the epicortex, ascospores and conidia shape and size, and amphithecial retrorse rhizines). The results conclusively reveal that: (1) there is not one single species but four separate species in the Mediterranean or sub Mediterranean areas of the world. Parmelina quercina and Parmelina carporrhizans (Euroasiatic species), Parmelina coleae sp. nov. (North America) and Parmelina elixia sp. nov. (Australia); (2) largely debated P. carporrhizans is not a synonym of P. quercina but supported as a valid species circumscribed to Macaronesic relict sites; (3) the geographical isolation of the Australian population is correlated with a large genetic distance; (4) morphological characters (ascospores and conidial variability and thallus epicortex) correlate with the phylogenetic hypothesis; (5) the new or revalidated species within Parmelina quercina are not cryptic species but morphologically recognizable taxa.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 455–467.     

Microscale variation in alpine grasslands: AFLPs reveal a high level of genotypic diversity in Primula minima (Primulaceae)

The microscale variation and spatial genetic structure of the alpine plant species Primula minima L. was analysed using AFLPs. AFLP analysis based on three primer combinations and 123 fragments revealed no identical genotypes among the 86 studied samples from a 300 × 300 cm plot. Variation within the study plot was high: Nei's gene diversity was 0.22, Shannon's information index 0.33 and the percentage of polymorphic bands was 60.9. Cluster analysis revealed four main groups of genetically similar individuals and mapping these individuals resulted in a clear spatial pattern, with samples from the same group often located close together. The observed microscale structure was corroborated using a Mantel test, which revealed significant correlation of genetic and spatial distances, and by the results of a spatial autocorrelation analysis that indicated a high level of similarity between adjacent samples. An analysis of molecular variance revealed clear differentiation (18%) between the spatial groups. Overall gene flow within the plot was 1.11 and ranged from 0.33 between the spatially most distant groups to 2.33 between directly neighbouring groups. The extraordinary level of diversity detected in this study indicates an unexpectedly strong relevance of reproduction by seed for the species P. minima in alpine grasslands. The strong microscale variation suggests, however, that there is limited dispersal of seeds. Clonal reproduction is of subsidiary importance to sexual reproduction and seems to occur only over very small distances.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 549–556.     

Morphological, vocal and genetic divergence in the Cettia acanthizoides complex (Aves: Cettiidae)

We used morphological, vocal and molecular (one mitochondrial and two nuclear loci) data to re-evaluate the taxonomic status of the taxa acanthizoides , concolor , and brunnescens in the Cettia acanthizoides (J. Verreaux, 1871) complex. We conclude that all three are valid taxa, and that acanthizoides of China and concolor of Taiwan are best treated as conspecific, whereas brunnescens of the Himalayas is better considered as a separate species. The degree of morphological, vocal, and genetic differentiation is variably congruent among all taxa; the recently separated acanthizoides and concolor differ slightly in plumage and structure but are indistinguishable in vocalizations, whereas the earlier diverged brunnescens and acanthizoides/concolor differ only slightly more in morphology but to a much greater degree in vocalizations. We stress the essential nature of taxonomic revisions as a prerequisite for the biodiversity estimates required for conservation planning.  © 2007 The Linnean Society of London, Zoological Journal of the Linnean Society , 2007, 149 , 437–452.     

Systematics and biogeography of the widespread Neotropical gekkonid genus Thecadactylus (Squamata), with the description of a new cryptic species

The systematics of the widespread neotropical gekkonid genus Thecadactylus is investigated using a combination of morphological, morphometric and mtDNA (cytochrome b ) sequence data, forming a total evidence dataset. The analysis tackles the common yet complex problem of a widespread taxon consisting of one or more cryptic species that are difficult to diagnose using morphology alone. The data are analysed using both maximum parsimony and Bayesian inference. Virtually all analyses resolve a well-supported south-western Amazonian clade distinct from the remainder of the recognized T. rapicauda . The south-western Amazonian clade is not only robustly supported, but also exclusive, geographically coherent and sufficiently distinct to warrant specific recognition. The new species is diagnosable on the basis of molecular sequences that are 23.0–26.9% divergent from those of T. rapicauda , and morphological evidence. Bayesian inference analysis robustly resolves meaningful and repeatable patterns of relationship. The biogeography of Thecadactylus is interpreted in the context of its two constituent species, and difficulties of resolving systematic and biogeographical patterns in widespread, cosmopolitan taxa are discussed. © 2007 The Linnean Society of London, Zoological Journal of the Linnean Society , 2007, 149 , 339–370.     

Cryptic speciation in liverworts – a case study in the Aneura pinguis complex

Bryophytes are amongst the most ancestral terrestrial plants and often have large distribution ranges across continents. Recent biochemical and molecular studies have suggested that many worldwide morphological species of bryophyte may represent genetically divergent and reproductively isolated cryptic species. We tested the cryptic species hypothesis in the thalloid liverwort Aneura pinguis complex. We applied analyses of chloroplast DNA (cpDNA) sequence variation and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods to discriminate between specimens of A. pinguis derived from various habitats in five distant geographical regions in Poland. Of the 19 specimens sequence characterized for the cpDNA tRNA^Leu region, seven haplotypes were identified divided into three nonmonophyletic clusters. The application of developed PCR-RFLP markers confirmed the existence of three tRNA^Leu types of A. pinguis (A–C) within the specimens derived from 21 populations. Sympatric populations of different tRNA^Leu types were found in lowland and mountain regions. No clear correlation between stand type and the presence of two tRNA^Leu types (A, B) was observed, as both were growing on soil, humus, and rocks. The tRNA^Leu type C was found only on humus and its distribution was restricted to low-lying northern populations. The above results indicate that the A. pinguis complex is highly differentiated at the molecular level and may represent three cryptic species.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 273–282.     

Flies on thistles: support for synchronous speciation?

Synchronous speciation of hosts and herbivorous insects predicts a congruent topology of host and insect phylogenies and similar evolutionary ages of host and insect taxa. To test these predictions for the specialized herbivorous fly genus Urophora (Diptera: Tephritidae), we used three different approaches. (i) We generated a phylogenetic tree of 11 European Urophora species from allozyme data and constructed a phylogeny of their hosts from published sources. Superimposing the Urophora tree on the host-plant tree we found no evidence for general congruence. (ii) We correlated genetic distances (Nei distances) of the host plants vs. the genetic distances of associated Urophora species. Overall, the relationship was not positive. Nevertheless, for some pairs of Urophora species and host plants genetic distances were in the same order of magnitude. (iii) We collected allozyme data for pairs of thistle taxa and pairs of herbivores on thistles together with independent time estimates. With these data we calibrated a molecular clock. There was a non-linear relationship between phylogenetic age and genetic distance, rendering the dating of deep events in thistle–insect evolution difficult. Nevertheless the derived molecular clock showed that the split of insect taxa lagged behind the split of hosts.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 775–783.     

Temporal patterns of diversity: assessing the biotic and abiotic controls on ant assemblages

In this study, we use 12 months of data from 11 ant assemblages to test whether seasonal variation in ant diversity is governed by either the structuring influences of interspecific competition or environmental conditions. Because the importance of competition might vary along environmental gradients, we also test whether the signature of competition depends on elevation. We find little evidence that competition structures the seasonal patterns of activity in the ant assemblages considered, but find support for the effects of temperature on seasonal patterns of diversity, especially at low-elevation sites. Although, in general, both competition and the environment interact to structure ant assemblages, our results suggest that environmental conditions are the primary force structuring the seasonal activity of the ant assemblages studied here.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 191–201.     

Low population genetic structuring of two cryptic bat species suggests their migratory behaviour in continental Europe

Although two cryptic pipistrelle bat species, Pipistrellus pipistrellus and Pipistrellus pygmaeus , belong among the most common bat species in Europe, it is still unclear whether they can migrate over long distances between summer and winter roosts. Long-distance migratory species may be expected to show low levels of genetic structuring in large areas due to regular mixing of the gene pool by mating that occurs during migration and/or hibernation. Conversely, the dispersal of gametes in sedentary species is spatially restricted, populations are more genetically structured, and isolation by relatively short distance is visible. By analysing diversity of highly variable microsatellites within and among summer colonies of both studied species in central Europe, we found that differentiation between populations is very weak. Both classical F _ST and Bayesian clustering approach failed to detect genetic structure among colonies and there was no significant isolation-by-distance pattern. The analyses of relatedness, however, revealed that individuals within colonies are more related than random suggesting philopatry of at least one sex. The results were very similar for the two species. The high level of gene flow among central European populations, even on large geographic distances, is discussed in relation with migrations, dispersal, and mating behaviour.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 103–114.     

Conflicting patterns of genetic and morphological variation in European Gentianella section Gentianella

Genetics in European Gentianella Moench sect. Gentianella were investigated using internal transcribed spacer (ITS) nrDNA, trn L-F cpDNA and Amplified Fragment Length Polymorphisms (AFLP) fingerprint data to analyse relationships within the section. Both a cladistic and a phenetic approach, as well as nonhierarchical analysis (PCA) and partitioning of molecular variance (AMOVA), were used. Sequence data did not resolve groups within the section. They reflected, however, geographical isolation between central European and eastern Carpathian ( G. lutescens and G. bulgarica ) as well as Caucasian samples ( G. caucasea ). AFLP data revealed a strong differentiation within G. germanica between populations of the Alps and north-western middle Europe, although there is hardly any morphological differentiation of taxonomic significance. Morphological similarities between G. aspera and G. pilosa were not reflected in genetics, and no support was found for G. aspera . Genetic similarities were not in line with morphology in G. lutescens and G. bulgarica . AFLP data only partly resolve relationships among taxa. High genetic similarities, as well as partitioning of molecular variance among taxa/populations of eastern middle Europe ( G. austriaca , G. bohemica , G. germanica , G. stiriaca ) and intermediate morphological characters in G. bohemica and G. stiriaca , might indicate reticulate evolution. The overall low sequence divergence within the section points to a very young origin of all taxa.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 175–187.     

Ecomorphological and genetic divergence between lowland and montane forms of the Pieris napi species complex (Pieridae, Lepidoptera)

The present study aimed to investigate the morphological and genetic differentiation of lowland and montane populations of the Pieris napi species complex in Norway and to make inferences about the subspecific status of Pieris napi adalwinda and Pieris napi napi . We measured 22 morphological characters on 228 individuals from eight populations in Central Norway (20–1100 m a.s.l.). In addition, lowland and mountain animals were reared at a high altitude locality. Half the animals from either locality were reared on mountain plant, and the other half on a lowland plant. Finally, a fragment of the mitochondrial cytochrome b gene was sequenced for individuals from Central and South-eastern Norway and Germany. Principal component analysis of morphological characters showed a zone of abrupt change from lowland to mountain morphological character states between populations at 506 m and 730 m a.s.l., respectively. The transplant experiment showed that the morphological differences have a genetic basis and that food plants have no impact on morphology. Limited, but significant, molecular genetic differentiation was found between lowland and mountain animals, but the phylogenetic analysis, however, showed that the lowland form ( P. n. napi ) is paraphyletic and the montane form ( P. n. adalwinda ) is monophyletic. Further study is required before taxonomic recognition can be applied.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 727–745.     

Divergent walleye (Sander vitreus)-mediated inducible defenses in the centrarchid pumpkinseed sunfish (Lepomis gibbosus)

Predation has important ecological and evolutionary consequences. Evolutionary responses to diversifying selection include genetic differentiation, the evolution of adaptive phenotypic plasticity, and the genetic differentiation of plastic responses between populations. We tested if pumpkinseed sunfish ( Lepomis gibbosus ) respond to predation cues by changing their external body form in functionally sensible ways. We then asked whether predation has influenced the divergence of coexisting littoral and pelagic ecomorphs, by testing for divergent predator-induced responses. Juvenile L. gibbosus of both ecomorphs were reared with and without predation cues supplied by walleye ( Sander vitreus ) feeding on L. gibbosus . Predation cues stimulated increased body depth and dorsal spine length, but no increase in anal spine length or pectoral fin size. The dorsal spines of pelagic ecomorphs also grew longer than did those of littoral ecomorphs, while positive body depth responses were similar in both ecomorphs. This is the second fish taxa in which predator-induced morphological responses have been found, and the first in which divergent responses have been detected between ecomorphs. This suggests that the developmental systems of L. gibbosus ecomorphs have diverged under selection related to predation. We propose that other 'resource polymorphisms' in fishes have evolved under selection arising from a variety of factors, including predation, and not just selection related to resource use.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 25–36.     

Amplified fragment length polymorphism genetic fingerprinting challenges the taxonomic status of the near-endemic species Chara curta Nolte ex Kütz. (Characeae)

Amplified fragment length polymorphism (AFLP) genetic fingerprinting of 14 accessions of Chara curta and Chara aspera Willd., sampled across a range of habitats and morphologies in Britain, suggests that these taxa are part of the variation within a single species complex. Two primer combinations generating 397 fragments (97% of which were polymorphic), analysed by Jaccard's similarity coefficient and principal co-ordinate analysis, did not recover groups which reflect the current taxonomy. By contrast with the genetic study, a Gower general similarity coefficient and principal co-ordinate analysis of 52 morphological characters recovered the currently recognized species groups. A Mantel test showed no significant correlation between the genetic data and the morphological data, supporting the hypothesis that phenotypic variability in Chara L. is either to some extent environmentally induced or represents developmental stages. Implications for the conservation status of C. curta in Britain are discussed.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 467–476.     

The robustness of geometric morphometrics in testing the morphological equivalence hypothesis among cichlid species from East African Great Lakes

As part of an ongoing broader investigation into whether similar morphologies among cichlid species from East African Great Lakes are due to common ancestry or convergent evolution, we used geometric morphometrics to quantitatively test the hyphothesis that Lake Tanganyika's Tropheus spp. is morphologically similar to Lake Malawi's Pseudotropheus spp. and Labeotropheus fuelleborni . Landmark-based techniques were used to investigate body shape variation and the results revealed highly significant differences among these genera with the exception of L. fuelleborni vs. Pseudotropheus ( Tropheops ) spp. An ordination of all specimens along the first two canonical variate (CV) axes indicated clear interlake separation, especially along the CV I axis, whereby Tropheus moori showed a dorsoventral expansion of the mid body section with a steeper forehead relative to Lake Malawi species. Pseudotropheus ( Maylandia ) zebra was discriminated from the rest primarily on the basis of its terminally positioned gape, which differed from the inferior subterminal gape that was prevalent in the other species. Thus, the present study reveals a pattern of parallel evolution between L. fuelleborni and Pseudotropheus ( Tropheops ) spp., but does not support an interlake convergent evolution hypothesis. Contrary to previous researchers' assertions, these results emphasize the robustness of geometric morphometrics when testing morphological equivalence hypothesis among cichlid species.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 1–9.     

Hidden Mediterranean biodiversity: molecular evidence for a cryptic species complex within the reef building vermetid gastropod Dendropoma petraeum (Mollusca: Caenogastropoda)

The reef building vermetid gastropod Dendropoma petraeum inhabits the warmest waters of the Mediterranean Sea and is considered a threatened marine species. The aim of this study was to characterize its genetic structure throughout its whole distribution range using mitochondrial and nuclear sequence data. Because of its sessile adult lifestyle and lack of a pelagic larval stage, we expected a markedly subdivided population structure with limited levels of gene flow. Fragments of the mitochondrial genes cytochrome c oxidase subunit I (COI) and 16S rRNA (16S), were sequenced, along with the nuclear ribosomal cluster (internal transcribed spacer; ITS) in specimens from 18 localities. Our analyses identified four highly distinct phylogroups separated by a mean divergence of > 14% according to the COI sequence data or > 9% according to 16S, but differing only slightly in morphology. The nuclear data (ITS) indicated a lower substitution rate (divergence among groups of around 1%). These large genetic distances among the four lineages clearly point to the existence of a cryptic species complex within D. petraeum comprising at least four species. Differences in the characteristics of intracapsular larval development and protoconch were also detected among these lineages. The allopatric distribution of these cryptic species supports a predominantly vicariant-based cladogenetic pattern for the genus Dendropoma in the Mediterranean.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 898–912.