Search

The authors used museomics to reconstruct the mitochondrial genome from two individuals of the Moroccan, endemic and extinct trout, Salmo pallaryi. They further obtained partial data from 21 nuclear genes previously used for trout phylogenetic analyses. Phylogenetic analyses, including publicly available data from the mitochondrial control region and the cytochrome b gene, and the 21 nuclear genes, place S. pallaryi among other North African trouts. mtDNA places S. pallaryi close to Salmo macrostigma within a single North African clade. Although the nuclear coverage of the genome was low, both specimens were independently positioned as sisters to one of two distantly related North African clades, viz. the Atlas clade with the Dades trout, Salmo multipunctatus. Phylogenetic discordance between mtDNA and nuclear DNA phylogenies is briefly discussed. As several specimens that were extracted failed to produce DNA of sufficient quality, the authors discuss potential reasons for the failure. They suggest that museum specimens in poor physical condition may be better for DNA extraction compared to better-preserved ones, possibly related to the innovation of formalin as a fixative before ethanol storage in the early 20th century.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Despite its importance in intertidal and shallow-water marine ecosystems, Caprella penantis continues to be one of the most taxonomically challenging amphipods in the world. A recent molecular study focusing on C. penantis sensu stricto pointed out the existence of three highly divergent lineages, indicating the possible existence of a process of ongoing speciation and, thus, casting doubt on the taxonomic status of this species. In the present study, we used an integrative approach to continue to shed light on the taxonomy and distribution of this caprellid. To this end, we combined morphological and genetic data (COI and 18S) and included, for the first time, populations from its type locality. Our analyses provide strong evidence of the existence of potentially three distinct species, genetically and geographically restricted, within C. penantis sensu stricto, with the distribution of the true C. penantis sensu stricto restricted to the UK (type locality), the northern coast of the Iberian Peninsula, and the Azores. Results show the co-occurrence of two of these species in a locality of northern Portugal and indicate the existence of distinct evolutionary and diversification patterns along the eastern Atlantic region. Overall, our study highlights the use of an integrative approach to properly assess species boundaries and unravel hidden biodiversity in amphipods.

Rabbitfish (Siganidae) are coral reef fish that are distributed across diverse habitats that include estuaries, mangroves, reefs, and even seaweed mats. Given their ecological diversity and natural widespread distributions across the Indo-Pacific region, we were interested to investigate the evolutionary history of this group and patterns of divergence that have contributed to their present-day distributions. In the present study, samples were collected from the South China Sea to study taxonomic and phylogenetic relationships, and divergence times. We investigated the taxonomic relationships among modern rabbitfish species, reconstructed their molecular phylogeny, and estimated divergence times among selected lineages based on a fragment of the mtDNA cytochrome oxidase I (COI) and sequences of the nuclear rhodopsin retrogene (RHO). Our results indicate that modern rabbitfish likely originated in the Indo-West Pacific during the late Eocene [37.4 million years ago (mya)], following which they diverged into three major clades during the Pliocene/Pleistocene. Subsequent diversification and origins of the majority of siganids may likely be associated with episodes of paleo-oceanographic events, including greenhouse and glaciation events (Eocene-Miocene) as well as major plate tectonic events (Pliocene-Pleistocene). Some modern siganid species may naturally hybridize with congeneric species where their geographical ranges overlap. A comprehensive taxonomic analysis revealed that the phylogeny of Siganidae (cladogenesis of Clades I, II, and III) is characterized by divergence in several external morphological characters and morphometric parameters. Our study demonstrates that morphological characteristics, geographical heterogeneity, and environmental change have contributed to siganids' historical diversification.

Environmental DNA (eDNA) provides a promising supplement to traditional sampling methods for population genetic inferences, but current studies have almost entirely focused on short mitochondrial markers. Here, we develop one mitochondrial and one nuclear set of target capture probes for the whale shark (Rhincodon typus) and test them on seawater samples collected in Qatar to investigate the potential of target capture for eDNA-based population studies. The mitochondrial target capture successfully retrieved similar to 235x (90x - 352x per base position) coverage of the whale shark mitogenome. Using a minor allele frequency of 5%, we find 29 variable sites throughout the mitogenome, indicative of at least five contributing individuals. We also retrieved numerous mitochondrial reads from an abundant nontarget species, mackerel tuna (Euthynnus affinis), showing a clear relationship between sequence similarity to the capture probes and the number of captured reads. The nuclear target capture probes retrieved only a few reads and polymorphic variants from the whale shark, but we successfully obtained millions of reads and thousands of polymorphic variants with different allele frequencies from E. affinis. We demonstrate that target capture of complete mitochondrial genomes and thousands of nuclear loci is possible from aquatic eDNA samples. Our results highlight that careful probe design, taking into account the range of divergence between target and nontarget sequences as well as presence of nontarget species at the sampling site, is crucial to consider. eDNA sampling coupled with target capture approaches provide an efficient means with which to retrieve population genomic data from aggregating and spawning aquatic species.

Environmental DNA (eDNA) provides a promising supplement to traditional sampling methods for population genetic inferences, but current studies have almost entirely focused on short mitochondrial markers. Here, we develop one mitochondrial and one nuclear set of target capture probes for the whale shark (Rhincodon typus) and test them on seawater samples collected in Qatar to investigate the potential of target capture for eDNA-based population studies. The mitochondrial target capture successfully retrieved similar to 235x (90x - 352x per base position) coverage of the whale shark mitogenome. Using a minor allele frequency of 5%, we find 29 variable sites throughout the mitogenome, indicative of at least five contributing individuals. We also retrieved numerous mitochondrial reads from an abundant nontarget species, mackerel tuna (Euthynnus affinis), showing a clear relationship between sequence similarity to the capture probes and the number of captured reads. The nuclear target capture probes retrieved only a few reads and polymorphic variants from the whale shark, but we successfully obtained millions of reads and thousands of polymorphic variants with different allele frequencies from E. affinis. We demonstrate that target capture of complete mitochondrial genomes and thousands of nuclear loci is possible from aquatic eDNA samples. Our results highlight that careful probe design, taking into account the range of divergence between target and nontarget sequences as well as presence of nontarget species at the sampling site, is crucial to consider. eDNA sampling coupled with target capture approaches provide an efficient means with which to retrieve population genomic data from aggregating and spawning aquatic species.

A phylogenetic analysis on specimen level was made in possible support of a multivariate analysis of the Calamus javensis complex. Nine species, at some time recognized within the complex, and several recognisable forms were included. The phylogenetic markers used were the nuclear 5S spacer (5S nrDNA) and the chloroplast Maturase K (matK). The Bayesian analysis showed that only 5S provided some resolution. The 50 % majority rule consensus showed one major polytomy with a few supported groups, which were mainly morphologically unsupported pairs of specimens. However, one group, the form C. tenompokensis (the only distinct group in a multivariate analysis) is morphologically distinct and phylogenetically monophyletic and can be recognized as a species. Of all other recognizable forms, we only consider C. acuminatus to be regarded as a variety as it was not supported in the morphometric analysis.

Bithynids snails are a widespread group of molluscs in European freshwater systems. However, not much information is available on trematode communities from molluscs of this family. Here, we investigate the trematode diversity of Bithynia tentaculata, based on molecular and morphological data. A total of 682 snails from the rivers Lippe and Rhine in North Rhine-Westphalia, Germany, and 121 B. tentaculata from Curonian Lagoon, Lithuania were screened for infections with digeneans. In total, B. tentaculata showed a trematode prevalence of 12.9% and 14%, respectively. The phylogenetic analyses based on 55 novel sequences for 36 isolates demonstrated a high diversity of digeneans. Analyses of the molecular and morphological data revealed a species-rich trematode fauna, comprising 20 species, belonging to ten families. Interestingly, the larval trematode community of B. tentaculata shows little overlap with the well-studied trematode fauna of lymnaeids and planorbids, and some of the detected species (Echinochasmus beleocephalus and E. coaxatus) constitute first records for B. tentaculata in Central Europe. Our study revealed an abundant, diverse and distinct trematode fauna in B. tentaculata, which highlights the need for further research on this so far understudied host–parasite system. Therefore, we might currently be underestimating the ecological roles of several parasite communities of non-pulmonate snail host families in European fresh waters

Bithynids snails are a widespread group of molluscs in European freshwater systems. However, not much information is available on trematode communities from molluscs of this family. Here, we investigate the trematode diversity of Bithynia tentaculata, based on molecular and morphological data. A total of 682 snails from the rivers Lippe and Rhine in North Rhine-Westphalia, Germany, and 121 B. tentaculata from Curonian Lagoon, Lithuania were screened for infections with digeneans. In total, B. tentaculata showed a trematode prevalence of 12.9% and 14%, respectively. The phylogenetic analyses based on 55 novel sequences for 36 isolates demonstrated a high diversity of digeneans. Analyses of the molecular and morphological data revealed a species-rich trematode fauna, comprising 20 species, belonging to ten families. Interestingly, the larval trematode community of B. tentaculata shows little overlap with the well-studied trematode fauna of lymnaeids and planorbids, and some of the detected species (Echinochasmus beleocephalus and E. coaxatus) constitute first records for B. tentaculata in Central Europe. Our study revealed an abundant, diverse and distinct trematode fauna in B. tentaculata, which highlights the need for further research on this so far understudied host–parasite system. Therefore, we might currently be underestimating the ecological roles of several parasite communities of non-pulmonate snail host families in European fresh waters

A phylogenetic analysis on specimen level was made in possible support of a multivariate analysis of the Calamus javensis complex. Nine species, at some time recognized within the complex, and several recognisable forms were included. The phylogenetic markers used were the nuclear 5S spacer (5S nrDNA) and the chloroplast Maturase K (matK). The Bayesian analysis showed that only 5S provided some resolution. The 50 % majority rule consensus showed one major polytomy with a few supported groups, which were mainly morphologically unsupported pairs of specimens. However, one group, the form C. tenompokensis (the only distinct group in a multivariate analysis) is morphologically distinct and phylogenetically monophyletic and can be recognized as a species. Of all other recognizable forms, we only consider C. acuminatus to be regarded as a variety as it was not supported in the morphometric analysis.

The designation of taxonomic units has important implications for the understanding and conservation of biodiversity. Eurasian vipers are a monophyletic group of viperid snakes (Serpentes, Viperinae), currently comprising four genera (Daboia, Macrovipera, Montivipera and Vipera) and up to 40 species. Taxonomic units have been described using a wide variety of methods and criteria, and consequently, considerable controversy still surrounds the validity of some currently listed species. In order to promote a consensus- A nd evidence-based taxonomy of Eurasian vipers, we analysed published mitochondrial and nuclear DNA sequences for this group to reconstruct phylogenetic relationships among currently recognized viper species. We also compiled information on external morphology to assess their morphological distinctiveness. Phylogenetic inference based on mtDNA sequences shows contrasting levels of divergence across genera and species and identifies several instances of non-monophyly in described species. Nuclear DNA sequences show extremely low levels of genetic variation, with a widespread pattern of allele sharing among distant species, and even among genera. Revision of morphological data shows that most species designations rely on scalation traits that overlap extensively among species of the same genus. Based on our combined assessment, we recognize 15 taxa as valid species, three taxa which likely represent species complexes, 17 taxa of doubtful validity as species, and five taxa for which species status is maintained but further research is highly recommended to assess taxonomic arrangements. We stress the need to implement integrative taxonomic approaches for the recognition of evidence-based taxonomic units in Eurasian vipers.

In the past 20 years, the number of subterranean taxa discovered in Australia, especially in the Pilbara bioregion, has considerably increased due to incidental environmental surveys often associated with mining development. Bathynellidae are an important component of stygofauna and they occur in most Australian aquifers, but their collection and identification are difficult due to their habitat, and small and fragile bodies with conservative morphology. The study of Pilbaranella ethelensis in the upper Fortescue catchment contributed to a better understanding of the group at local scale, but knowledge at larger catchment scale is still limited. Abundant material collected by different environmental consultant companies on behalf of mining companies allowed an accurate analysis of the populations of Central Hamersley Range bathynellids. A new genus and one new species from the lower Fortescue catchment, Fortescuenella serenitatis gen. et sp. nov., is described using an approach that integrates morphological and molecular data. Three additional lineages are defined through morphology and DNA sequencing, using Automatic Barcode Gap Discovery and Poisson Tree Processes species delimitation methods. Based on previous knowledge on bathynellids and other stygofauna we expected to find multiple taxa, geographically restricted, possibly related to each other and to Pilbaranella genus described upstream of the Fortescue catchment. The phylogenetic reconstruction of the relationships among known lineages of the family in the Pilbara highlights a pattern of distribution characterized by a complex evolutionary history that does not reflect contemporary surface water catchments, suggesting a diversification that preceded the aridification processes started in the north of Western Australia during the Miocene. This study also clarifies the status of the 'cosmopolitan' Bathynella by excluding the Australian bathynellids from this genus.

Mitochondrial DNA testing is expanding the use of DNA as a forensic tool for human identification. The testing of mitochondrial DNA is a common practice for biological samples, which are compromised, degraded and contain limited STR information. In these cases, the amplification and sequencing of mitochondrial DNA becomes a valuable tool in determining the source of DNA samples. A review of mitochondrial DNA testing in forensic science reveals a number of improvements which can be made to this process. The project therefore aimed to improve the process of mitochondrial DNA testing. In particularly, it focuses on the developing and testing of mitochondrial DNA markers involved in the quantification and amplification of samples. The quantification of mitochondrial DNA is important to optimising the amplification of DNA samples. Methods for quantification of mitochondrial DNA commonly involve estimating mitochondrial DNA quantities from nuclear DNA. This is usually unreliable as a high variability of mitochondrial DNA copy number exists between human cells. Hence, in this study, the development of a specific DNA assay for mitochondrial DNA provides a reliable determination of mitochondrial copy number. The testing of this novel assay has shown it is specific, sensitive and reproducible in DNA samples of artificially degraded qualities. As such, the incorporating of the DNA assay into workflows of mitochondrial DNA testing will improve the overall amplification of samples for downstream processes such as mitochondrial DNA sequencing. Furthermore, the transition of mitochondrial DNA sequencing from capillary electrophoresis to massively parallel sequencing platforms is increasing the feasibility of typing multiple DNA fragments in a single reaction. This has led to the development of small PCR markers, which are capable of amplifying the entire mitochondrial genome even in challenging forensic samples. While the sequences of these PCR markers and panels are available for

Temperate terrestrial species in Europe were hypothesized to have been restricted to southern peninsular refugia (Iberia, Italy, Balkans) during the height of the last glacial period. However, recent analyses of fossil evidence indicate that some temperate species existed outside these areas during the last glacial maximum (LGM). Red foxes (Vulpes vulpes) in particular, could have been distributed across the southern half of the continent, potentially forming one continuous population. To investigate these hypotheses, we used 21 nuclear microsatellite loci and two fragments (768 bp) of mitochondrial DNA to characterize the population structure among a continent-wide sample of 288 European red foxes. We tested whether European red foxes clustered into discrete populations corresponding to the hypothetical peninsular refugia. Additionally, we sought to determine if distinct northern populations were formed after post-glacial recolonization. Our results indicated that only the foxes of Iberia appeared to have remained distinct over a considerable period of time (32–104 kya). Spanish red foxes formed their own genotypic cluster; all mtDNA haplotypes were endemic and closely related, and together both the mitochondrial and nuclear datasets indicated this population contributed little to postglacial recolonization of Northern Europe. In contrast, red foxes from Italy and the Balkans contributed significantly to, or were part of, a wider, admixed population stretching across mid-latitude Europe. In Northern Europe, we identified a Scandinavian population that had an ancestral relationship with red foxes to the south, and a more recent relationship with those to the east, in Russia. We also resolved two distinct populations on the islands of Ireland and Britain that had been separated from one another, and from those on the continent, since the late Pleistocene/mid Holocene (∼4–24 kya).

Mitochondrial DNA testing is expanding the use of DNA as a forensic tool for human identification. The testing of mitochondrial DNA is a common practice for biological samples, which are compromised, degraded and contain limited STR information. In these cases, the amplification and sequencing of mitochondrial DNA becomes a valuable tool in determining the source of DNA samples. A review of mitochondrial DNA testing in forensic science reveals a number of improvements which can be made to this process. The project therefore aimed to improve the process of mitochondrial DNA testing. In particularly, it focuses on the developing and testing of mitochondrial DNA markers involved in the quantification and amplification of samples. The quantification of mitochondrial DNA is important to optimising the amplification of DNA samples. Methods for quantification of mitochondrial DNA commonly involve estimating mitochondrial DNA quantities from nuclear DNA. This is usually unreliable as a high variability of mitochondrial DNA copy number exists between human cells. Hence, in this study, the development of a specific DNA assay for mitochondrial DNA provides a reliable determination of mitochondrial copy number. The testing of this novel assay has shown it is specific, sensitive and reproducible in DNA samples of artificially degraded qualities. As such, the incorporating of the DNA assay into workflows of mitochondrial DNA testing will improve the overall amplification of samples for downstream processes such as mitochondrial DNA sequencing. Furthermore, the transition of mitochondrial DNA sequencing from capillary electrophoresis to massively parallel sequencing platforms is increasing the feasibility of typing multiple DNA fragments in a single reaction. This has led to the development of small PCR markers, which are capable of amplifying the entire mitochondrial genome even in challenging forensic samples. While the sequences of these PCR markers and panels are available for

The European lantern fly, Dictyophara europaea, is an alternative vector of the Flavescence doree phytoplasma (FDp) disease of grapevine in European vineyards, enabling infection initiation from wild reservoir compartment (Clematis vitalba). Heretofore recorded rate of D. europaea FDp-infection has been very low (3%), making it less epidemiologically significant than would be expected based on reservoir plant infection rate (30%). In this study we present findings on a heavily FDp-infected D. europaea population ( gt 60%), on the natural Wolbachia infection of populations with low FDp-infection rates (DeWo+) and on Wolbachia absence in highly FDp-infected population (DeWo-). We examine several possible causes underlying the differences in vector infection rates: (a) population genetic characteristics of D. europaea and correlation with Wolbachia strain wEur natural infections, (b) Wolbachia effects on fitness components of DeWo+ laboratory colony and (c) rate of reservoir plant FDp-infection and differences in FDp genotypes harboured by low and highly infected vector populations. The vector genetic diversity level was found to be lower in DeWo+ than in uninfected individuals and to exhibit a different evolution of fixed haplotypes. All DeWo+ populations were infected with the same strain of wEur. The FDp was found to be genetically diversified (five genotypes) but had no relation to infection rates. We did not find evidence of fitness upgrades with regard to Wolbachia infection status. Although more experimentation is needed, it seems that Wolbachia confers protection against FDp or is in competition with FDp according to the observed correlations: low FDp-infected vector populations are infected with Wolbachia and vice versa.

Despite extensive investigation, a complete understanding of the evolutionary history of the Macropodidae (kangaroos and their kin) has remained elusive. This research has utilized DNA sequences and retrotransposons (genes that jump around within the genome) to shed light on the evolutionary timing and dynamics of these iconic marsupials over the past 20 million years, and draw correlations with past climate change events. The research shows that these marsupials underwent a rapid radiation, diversifying into a wide array of forms, coincident with a trend of climatic cooling and drying over the past ~8 million years.

Three species of the genus Asterina are known to inhabit the Mediterranean Sea and the north-eastern Atlantic Ocean: Asterina gibbosa (Pennant, 1777), A. pancerii (Gasco, 1870) and A. phylactica Emson & Crump, 1979. Differentiation of these species has primarily been based only on subtle characters (some highly debatable), such as colour or size. Therefore, this study aimed to review the morphological data characterising members of the genus, to incorporate new characters that may clarify morphological analyses and to couple morphological data with molecular evidence of differentiation based on the analysis of partial sequences of the cytochrome c oxidase subunit I (COI) and 18S rDNA (18S) genes and two anonymous nuclear loci (AgX2 and AgX5). The different lineages and cryptic species identified from the molecular analysis were then morphologically characterised, which was challenging given the limited number of diagnostic characters. Two of the five monophyletic lineages obtained molecularly (COI divergence >4%), further supported by differences in morphological characters and reproductive behaviour, are proposed as new species: Asterina martinbarriosi, sp. nov. from the Canary Islands, Spain (eastern central Atlantic Ocean) and Asterina vicentae, sp. nov. from Tarragona, north-eastern Spain (western Mediterranean Sea).

The European common lizard (Zootoca vivipara) is a widely distributed species across Europe and Asia exhibiting two reproductive modes (oviparity/viviparity), six major lineages and several sublineages. It has been used to tackle a large variety of research questions, nevertheless, few nuclear DNA sequence markers have been developed for this species. Here we developed 79 new nuclear DNA sequence markers using a clonation protocol. These markers were amplified in several oviparous and viviparous specimens including samples of all extant clades, to test the amplification success and their diversity. 49.4% of the markers were polymorphic and of those, 51.3% amplified in all and 94.9% amplified in 5–7 of the extant Z. vivipara clades. These new markers will be very useful for the study of the population structure, population dynamics, and micro/macro evolution of Z. vivipara. Cross-species amplification in four lizard species (Psammodromus edwardsianus, Podarcis muralis, Lacerta bilineata, and Takydromus sexlineatus) was positive in several of the markers, and six makers amplified in all five species. The large genetic distance between P. edwardsianus and Z. vivipara further suggests that these markers may as well be employed in many other species.

Three species of giant barrel sponge are currently recognized in two distinct geographic regions, the tropical Atlantic and the Indo-Pacific. In this study, we used molecular techniques to study populations of giant barrel sponges across the globe and assessed whether the genetic structure of these populations agreed with current taxonomic consensus or, in contrast, whether there was evidence of cryptic species. Using molecular data, we assessed whether giant barrel sponges in each oceanic realm represented separate monophyletic lineages. Giant barrel sponges from 17 coral reef systems across the globe were sequenced for mitochondrial (partial CO1 and ATP6 genes) and nuclear (ATPsβ intron) DNA markers. In total, we obtained 395 combined sequences of the mitochondrial CO1 and ATP6 markers, which resulted in 17 different haplotypes. We compared a phylogenetic tree constructed from 285 alleles of the nuclear intron ATPsβ to the 17 mitochondrial haplotypes. Congruent patterns between mitochondrial and nuclear gene trees of giant barrel sponges provided evidence for the existence of multiple reproductively isolated species, particularly where they occurred in sympatry. The species complexes in the tropical Atlantic and the Indo-Pacific, however, do not form separate monophyletic lineages. This rules out the scenario that one species of giant barrel sponge developed into separate species complexes following geographic separation and instead suggests that multiple species of giant barrel sponges already existed prior to the physical separation of the Indo-Pacific and tropical Atlantic.

Development of 34 new and multiplexing of seven existing microsatellite loci for Zootoca vivipara (Squamata: Lacertidae). Few microsatellite loci exist for the European common lizard Zootoca vivipara, a widely used model species for studying population dynamics, sexual selection, population genetics, parity evolution, and physiology. The existing primers did not amplify in all lineages, and multiplexes were not optimised. In this work, a total of 34 new polymorphic microsatellite markers have been developed for this species and tested in 64 specimens belonging to oviparous and viviparous clades (B and D). The microsatellites were combined into seven different multiplexes. Results showed that all but one loci successfully amplified in all samples and both clades. The number of alleles detected per locus ranged from seven to 22 alleles and the effective number from 1.58 to 7.82. The observed heterozygosity ranged from 0.312 to 0.930, showing that all loci were highly variable. Oviparous and viviparous clades exhibited significant genetic differences (in FST). In addition to these new markers, the 7 previously published and widely used microsatellite loci have been multiplexed and tested in oviparous clades. All these innovations will allow for time-saving and robust analyses in Zootoca vivipara, boosting evolutionary and population studies and easing paternity analyses.

Taxonomy of the live-bearing fish of the genus Ilyodon Eigenmann, 1907 (Goodeidae), in Mexico, is controversial, with morphology and mitochondrial genetic analyses in disagreement about the number of valid species. The present study accumulated a comprehensive DNA sequences dataset of 98 individuals of all Ilyodon species and mitochondrial and nuclear loci to reconstruct the evolutionary history of the genus. Phylogenetic inference produced five clades, one with two sub-clades, and one clade including three recognized species. Genetic distances in mitochondrial genes (cytb: 0.5%¿2.1%; coxI: 0.5%¿1.1% and d-loop: 2.3%¿10.2%) were relatively high among main clades, while, as expected, nuclear genes showed low variation (0.0%¿0.2%), with geographic concordance and few shared haplotypes among river basins. High genetic structure was observed among clades and within basins. Our genetic analyses, applying the priority principle, suggest the recognition only of Ilyodon whitei and Ilyodon furcidens, with I. cortesae relegated to an invalid species, the populations of which belong to I. whitei

The aim of this thesis is to investigate how discussions in archaeology and genetics influence the consensus on human origins and migrations in the South Pacific. By analyzing the genetic research on chicken- and sweet potato-DNA, I present a general overview of how genetics and archaeology shape the understanding of how humans have colonized the Pacific. By deconstructing a review on how the Pacific was settled based on aDNA, I analyze a geneticist’s perspective on archaeological problems. Through this analysis I suggest how archaeology should be approached on a theoretical level in order to be relevant in understanding human migrations in the Pacific. I propose that archaeology’s strength lie in interpreting material culture through an agency perspective in order to reach a dimension not obtainable by biological perspectives, Syftet med denna uppsats är att undersöka hur diskussioner i arkeologi och genetik påverkar hur vi ser på mänskliga migrationer i Oceanien. Genom att analysera den genetiska forskning som gjorts på kyckling och sötpotatis ges en övergripande bild av hur genetik och arkeologi formar den förståelse som finns för hur människan koloniserat Söderhavet. Genom att dekonstruera en sammanställning av den genetiska forskning som gjorts på mänskligt DNA i Oceanien analyseras en genetikers synsätt på arkeologiska problemställningar. Genom analysen i denna uppsats föreslår jag hur arkeologi borde arbeta på ett teoretiskt plan för att vara relevant i hur vi förstår Oceaniens migrationsmönster. Jag föreslår att arkeologins styrka ligger i att tolka den materiella kulturen genom ett agency-perspektiv för att komma åt en dimension av migrationsproblematiken som inte går att nås genom biologiska perspektiv.

The aim of this thesis is to investigate how discussions in archaeology and genetics influence the consensus on human origins and migrations in the South Pacific. By analyzing the genetic research on chicken- and sweet potato-DNA, I present a general overview of how genetics and archaeology shape the understanding of how humans have colonized the Pacific. By deconstructing a review on how the Pacific was settled based on aDNA, I analyze a geneticist’s perspective on archaeological problems. Through this analysis I suggest how archaeology should be approached on a theoretical level in order to be relevant in understanding human migrations in the Pacific. I propose that archaeology’s strength lie in interpreting material culture through an agency perspective in order to reach a dimension not obtainable by biological perspectives, Syftet med denna uppsats är att undersöka hur diskussioner i arkeologi och genetik påverkar hur vi ser på mänskliga migrationer i Oceanien. Genom att analysera den genetiska forskning som gjorts på kyckling och sötpotatis ges en övergripande bild av hur genetik och arkeologi formar den förståelse som finns för hur människan koloniserat Söderhavet. Genom att dekonstruera en sammanställning av den genetiska forskning som gjorts på mänskligt DNA i Oceanien analyseras en genetikers synsätt på arkeologiska problemställningar. Genom analysen i denna uppsats föreslår jag hur arkeologi borde arbeta på ett teoretiskt plan för att vara relevant i hur vi förstår Oceaniens migrationsmönster. Jag föreslår att arkeologins styrka ligger i att tolka den materiella kulturen genom ett agency-perspektiv för att komma åt en dimension av migrationsproblematiken som inte går att nås genom biologiska perspektiv.

Tardigrades represent one of the most abundant groups of Antarctic metazoans in terms of abundance and diversity, thanks to their ability to withstand desiccation and freezing; however, their biodiversity is underestimated. Antarctic tardigrades from Dronning Maud Land and Victoria Land were analysed from a morphological point of view with light microscopy and scanning electron microscopy, and from a molecular point of view using two genes (18S, 28S) analysed in Bayesian inference and maximum-likelihood frameworks. In addition, indel-coding datasets were used for the first time to infer tardigrade phylogenies. We also compared Antarctic specimens with those from Italy and Greenland. A combined morphological and molecular analysis led to the identification of two new evolutionary lineages, for which we here erect the new genera Acanthechiniscus, gen. nov. (Echiniscidae, Echiniscoidea) and Mesobiotus, gen. nov. (Macrobiotidae, Macrobiotoidea). Moreover, two species new to science were discovered: Pseudechiniscus titianae,sp. nov. (Echiniscidae : Echiniscoidea) and Mesobiotus hilariae, sp. nov. (Macrobiotidae : Macrobiotoidea). This study highlights the high tardigrade diversity in Antarctica and the importance of an integrated approach in faunal and taxonomic studies.

Integrating information from species occurrence data, environmental variables and molecular markers can provide valuable insights about the processes of population persistence and differentiation. In this study, we present the most comprehensive overview of the evolutionary history of the North African salamander Salamandra algira (Caudata, Salamandridae) to date, including analyses of climatic and topographical variables, and sequences of two mitochondrial and two nuclear DNA fragments, with a special focus on Algerian populations, under‐represented in previous studies. Coalescent‐based phylogenetic analyses of mtDNA data recover four well‐supported population groups corresponding to described subspecies, with a western clade including populations in north‐western Morocco (with two subclades corresponding to the subspecies tingitana and splendens), and an eastern clade including populations from north‐eastern Morocco (subspecies spelaea) and Algeria (subspecies algira). Inferred split times between major clades date back to the Miocene, with additional splits within each major clade in the Plio‐Pleistocene. Present climatic (aridity) and topographical factors account for geographical discontinuities across population groups and help identify potential areas of secondary contact between clades corresponding to the subspecies tingitana and splendens in the Rif mountains in Morocco. Niche analysis indicates the absence of phylogenetic signal in the use of environmental space in this species.

In this study, we used sequences of two mitochondrial genes, cytochrome c oxidase I (COI) and 16S rRNA, and one nuclear gene, 28S rRNA, to test the monophyly of the sea star genus Echinaster, and understand the phylogenetic relationships among species and subgenera within this genus. Phylogenetic analyses based on Bayesian inference and maximum likelihood methods revealed three clades with high values of genetic divergence among them (K2P distances for COI over 23%). One of the clades grouped all Echinaster (Othilia) species, and the other two clades included Echinaster (non-Othilia) species and Henricia species, respectively. Although the relationships among Henricia, Othilia, and Echinaster could not be completely clarified, the Othilia clade was a well-supported group with shared diagnostic morphological characters. Moreover, the approximately unbiased test applied to the phylogenetic reconstruction rejected the hypothesis of the genus Echinaster as a monophyletic group. According to these results, we suggest the revalidation of Othilia as a genus instead of a subgenus within Echinaster. Our study clarifies important points about the phylogenetic relationships among species of Echinaster. Other important systematic questions about the taxonomic classification of Echinaster and Henricia still remain open, but this molecular study provides bases for future research on the topic.

Niphanda fusca is an endangered myrmecophilous butterfly inhabiting environments at early stages of succession. Most of its habitats are places where succession is prevented by human activity. In some places, however, N. fusca lives in natural semi-bare areas, such as cliffs in mountains or grasslands around volcanos. We investigate the genetic structure of N. fusca in Japan and South Korea to address two questions. (1) Are populations in natural environments genetically different from those in secondary environments? and (2) Do populations in natural environments possess greater genetic diversity than those in secondary environments? The AMOVA results indicated that the populations in natural environments and those in secondary environments were differentiated to some extent; however, more than 80 % of genetic variation was found to occur within the same habitat type and within each population. We found no differences in genetic diversity between populations in the two environments. At present, we have not found a strong reason to consider populations in the two environments as different evolutionarily significant units. We think it is practical to conserve populations in natural environments at first, because in this case we need not manage habitats to protect N. fusca. We have only to inhibit habitat destruction. In contrast, in order to conserve populations in secondary environments, we would have to continue managing the habitats. This is far more difficult than inhibiting habitat destruction.

The power of phylogeographic inference resides in its ability to integrate information from multiple sources in an iterative hypothesis-testing framework. In this paper, we build upon previous mtDNA-based hypotheses about the evolutionary history of the Iberian newt Lissotriton boscai using sequences o

We interpreted the results of nuclear DNA sequencing to be inconsistent with the recognition of California Gnatcatcher (Polioptila californica) subspecies. McCormack and Maley (2015) suggested that our data did support 2 taxa, one of which was P. c. californica, listed as Threatened under the Endangered Species Act (ESA). We summarize here how 2 sets of researchers with access to the same data reached different conclusions by including different analyses. We included the southern subspecies’ boundary from the taxonomy of Atwood (1991), the taxonomic basis for the ESA listing, which resulted in an Analysis of Molecular Variance that provided no support for subspecies. In contrast, using a novel taxonomic hypothesis without precedent in the literature, McCormack and Maley (2015) found statistically significant FST values for 2 loci, which they suggested supports P. c. californica. We propose that our mitochondrial and nuclear data had sufficient power to capture geographical structure at either the phylogenetic (monophyly) or traditional ‘‘75% rule’’ level. McCormack and Maley (2015) suggested that finding an absence of population structure was a ‘‘negative result,’’ whereas we consider it to be the null hypothesis for a species with gene flow and no geographical barriers. We interpret the unstructured mtDNA and nuclear DNA trees, the STRUCTURE analysis supporting one group, the identification of just 26% (and not 75%) of individuals of P. c. californica with the most diagnostic nuclear locus, the overall GST that suggests that over 98% of the variation is explained by nontaxonomic sources, and the lack of evidence of ecological differentiation to indicate that P. c. californica is not a valid subspecies. McCormack and Maley (2015) suggest that statistically significant differences at 2 loci that explained ,6% of the genetic variation, and previous morphological data, support recognition of P. c. californica. If ornithology continues to recognize subspecies, these dif

The genus Rhynchocalamus comprises three species distributed in Southwest Asia. Little is known about them, most probably because of their secretive fossorial lifestyle. The poor knowledge of the genus is even underscored by the fact that its phylogenetic affinities remained unclear until very recently. The least known of the species, Rhynchocalamus arabicus, is known only from the holotype collected in Aden, Yemen, and it has not been observed since its description in 1933. Here we provide a second record for this species, which represents the first record of this genus for Oman. This extends its range in southern Arabia by more than 1000 km. The observed specimen was determined as R. arabicus on the basis of its similarity in size, color, and scalation with the holotype. Furthermore, we sequenced three mitochondrial (12S, 16S, cytb) and one nuclear (cmos) genes for R. arabicus and for two individuals of R. melanocephalus and one R. satunini and inferred the phylogenetic relationships of all currently recognized species of the genus for the first time. The results of our phylogenetic analyses indicate that Rhynchocalamus is a member of the Western Palearctic clade of Colubrinae and is sister to Lytorhynchus, with which it forms a very well supported clade and shares some morphological characters. As our results show, R. satunini is the basal lineage of the genus and R. melanocephalus is sister to R. arabicus.

Aim: Suture zones are areas where closely related species from different biogeographical regions come into contact and interbreed. This concept originated from the study of terrestrial ecosystems but it remains unclear whether a similar phenomenon occurs in the marine environment. Here we investigate a potential suture zone from a previously unknown hybrid hotspot at the Socotra Archipelago (Yemen), located in the Arabian Sea, where fauna from the Red Sea, Gulf of Aden, Arabian Sea, western Indian Ocean and greater Indo-Polynesian Province intersect. Location: Red Sea, Gulf of Aden, Arabian Sea and Indian Ocean. Methods: Putative hybrid reef fish were identified based on intermediate coloration and morphology. Underwater observations and collections were conducted to determine: (1) whether parent species form heterospecific social groups or breeding pairs; (2) the sex and reproductive status of morphologically intermediate individuals; and (3) whether parent species were forming mixed species associations owing to a dearth of conspecific partners. To support hybrid status, morphologically intermediate and parental individuals were genotyped using mitochondrial DNA cytochrome c oxidase subunit I (COI), nuclear recombination-activating gene 2 (RAG2) and the nuclear TMO-4C4 (TMO) gene.Results: We observed putative hybrids involving 14 species from four reef fish families at Socotra. Most cases involved a parental species with a restricted distribution (e.g. Red Sea or Arabian Sea) and a broadly distributed Indo-Pacific species. In most cases, at least one of the parent species was rare at Socotra. Hybrid gene flow was largely unidirectional, and although introgression was rare, we found evidence that some butterflyfish and surgeonfish hybrids were fertile and formed breeding groups with parental species. Main conclusions: The rate of hybrid discovery at Socotra is much greater than that recorded elsewhere in the marine environment and involved both allopatric and sympa

U ovom radu analizirana je fitohemijska i genetiţka varijabilnost i diferencijacija sedam autohtonih populacija Pinus nigra u Srbiji koje pripadaju razliţitim infraspecijskim taksonima (ssp. nigra, var. gocensis, ssp. pallasiana i var. banatica). U dvogodišnjim ţetinama 195 jedinki iz sedam populacija odreŤen je sadrţaj i varijabilnost n-alkana i terpena. Kutikularne voskove ţetina istraţivanih populacija crnog bora odlikuje serija nalkana od C16 do C33, osim u populacijama opisanim kao ssp. nigra u kojima je detektovana serija od C18 do C33. Najzastupljeniji su n-alkani sa neparnim brojem C atoma: C23, C25, C27 i C29 (proseţni sadrţaj 10.3, 16.5, 15.1 i 10.7%, respektivno). U etarskim uljima ţetina utvrŤeno je ukupno 75 komponenti, od koji je 58 identifikovano. Dominantne komponente su α-pinen (43.6%) i germakren D (29.8%), koje zajedno ţine 73.4% mase ulja. Multivariacione statistiţke analize (PCA, CDA i HCA) koriššenih fitohemijskih markera su, generalno, pokazale tendenciju formiranja tri grupe populacija (nigra, pallasiana i banatica). Na osnovu varijabilnosti terpena, populacije Priboj i Goţ (determinisane kao var. gocensis) pokazale su umerenu tendenciju odvajanja od prve grupe (ssp. nigra), dok je prema sastavu n-alkana populacija Priboj pokazala još vešu tendenciju odvajanja. Pretpostavljeno je da takson gocensis predstavlja tranzicionu formu izmeŤu taksona nigra na zapadu i pallasiana na jugoistoku Srbije. U cilju odreŤivanja genetiţkog diverziteta i diferencijacije sedam istraţivanih populacija P. nigra iz Srbije, testirano je 114 jedinki pomošu jedarnih (nuDNK), hloroplastnih (cpDNK) i mitohondrijalnih (mtDNK) markera koji imaju bi-parentalno, paternalno i maternalno nasleŤivanje, respektivno, kao i razliţitu brzinu evolucije. Zbog nedostatka jedarnih mikrosatelita (nuSSRs) specijalno razvijenih za P. nigra, uraŤena je inter-specijska PCR amplifikacija tri EST-SSRs razvijenih za P. taeda. Iako su dva ESTSSR lokusa uspešno PCR-amplifikovana u našim uzorci, In this work phytochemical and genetic variability and differentiation of seven native populations belonging to different infraspecific taxa of Pinus nigra (ssp. nigra, var. gocensis, ssp. pallasiana i var. banatica) in Serbia were analyzed. In the needles of 195 trees from seven populations, composition and variability of n-alkanes and terpenes were determined. The size of the n-alkanes found in the needle cuticular waxes of the black pine populations investigated ranged from C16 to C33, with the exception of that of the subspecies nigra, which ranged from C18 to C33. The most abundant n-alkanes were the four odd-numbered n-alkanes C23, C25, C27, and C29 (mean contents of 10.3, 16.5, 15.1, and 10.7%, respectively). In the needle essential oils a total of 58 individual components out of the 75 compounds detected could be identified. The major components of the oils were α-pinene (43.6%) i germakrene D (29.8%), comprising together in average 73.4% of the essential oil composition. Multivariate statistical analysis (PCA, CDA, and HCA) of used phytochemical markers showed, in general, a tendency to the formation of three populations groups (nigra, pallasiana, and banatica). Based on the terpene variability, populations Priboj and Goţ, determined as var. gocensis, showed a moderate tendency of separation from the first group (ssp. nigra), while based on n-alkane composition, population Priboj showed even a greater tendency of separation. It was assumed that taxon gocensis might be a transitional form between taxa nigra occurring in the west and pallasiana growing in the southeast of Serbia. We screened 114 trees from seven studied P. nigra populations from Serbia with nuclear (nrDNA), chloroplast (cpDNA) and mitochondrial (mtDNA) markers which have bi-parental, paternal and maternal inheritance, respectively, and different evolutionary rates to infer levels of genetic diversity and structuring within studied area. Due to the lack of nuclear microsatellites (nuSSRs) spec

We detail the evolutionary history of the desert woodrat complex (lepida group, genus Neotoma) of western North America. Our analyses include standard multivariate morphometrics of museum specimens coupled with mitochondrial and nuclear DNA sequences and microsatellite loci. We trace the spatial and temporal diversification of this group of desert dwelling rodents, revising species boundaries and delineate subspecies we consider valid.

The Hymenodicteae-Naudeeae clade is a predominantly Paleotropical group with 220 species in 28 genera. The phylogertetic relationships and generic limits within Naucleeae have previously been assessed using combined molecular-morphological data, however the status of some genera remains questionable. The evolutionary relationships within Hymenodictyeae have never been investigated before. We performed phylogenetic analyses of the Hymenodictyeae-Naucleeae clade using nuclear [nrETS; nrITS] and chloroplast [ndhF; rbcL; rps16; trnT-F] data and a large sampling of both tribes. Our study supports the monophyly of the tribes, all subtribes of Naucleeae (Adininae, Breoniinae, Cephalanthinae, Corynantheinae, Mitragyninae, Naucleinae, and Uncariinae), and the Hymenodictyeae genera Hymenodictyon and Paracorynanthe. In Naucleeae, the monotypic genera Adinauclea, Metadina, and Pertusadina are nested within Adina, Mitragyna within Fleroya, Ludekia, Myrmeconauclea, and Ochreinauclea within Neonauclea, and Burttdavya and Sarcocephalus within Nauclea. Corynanthe and Pausinystalia are mutually paraphyletic. We provisionally maintain the current generic status of Neonauclea and its allied genera, pending further study. In sum, we recognize 17 genera in Naucleeae: Adina s. l., Breonadia, Breonia, Cephalanthus, Corynanthe s. l., Diyaminauclea, Gyrostipula, Janotia, Khasiaclunea, Ludekia, Mitragyna s. l., Myrmeconauclea, Nauclea s. l., Neolamarckia, Neonauclea, Ochreinauclea, and Uncaria. Five new combinations were made: Adina euryncha, Adina malaccensis, Corynanthe lane-poolei subsp. iturense, Corynanthe talbotii, and Nauclea nyasica., AuthorCount:4

We detail the evolutionary history of the desert woodrat complex (lepida group, genus Neotoma) of western North America. Our analyses include standard multivariate morphometrics of museum specimens coupled with mitochondrial and nuclear DNA sequences and microsatellite loci. We trace the spatial and temporal diversification of this group of desert dwelling rodents, revising species boundaries and delineate subspecies we consider valid.

The genus Cheirogaleus, the dwarf lemurs, is a radiation of strepsirrhine primates endemic to the island of Madagascar. The dwarf lemurs are taxonomically grouped in the family Cheirogaleidae (Infraorder: Lemuriformes) along with the genera Microcebus, Mirza, Allocebus, and Phaner. The taxonomic history of the genus Cheirogaleus has been controversial since its inception due to a paucity of evidence in support of some proposed species. In this study, we addressed this issue by expanding the geographic breadth of samples by 91 individuals and built upon existing mitochondrial (cytb and COII) and nuclear (FIBA and vWF) DNA datasets to better resolve the phylogeny of Cheirogaleus. The mitochondrial gene fragments D-loop and PAST as well as the CFTR-PAIRB nuclear loci were also sequenced. In agreement with previous genetic studies, numerous deep divergences were resolved in the C. major, C. minor and C. medius lineages. Four of these lineages were segregated as new species, seven were identified as confirmed candidate species, and four were designated as unconfirmed candidate species based on comparative mitochondrial DNA sequence data gleaned from the literature or this study. Additionally, C. thomasi was resurrected. Given the widespread distribution of the genus Cheirogaleus throughout Madagascar, the methodology employed in this study combined all available lines of evidence to standardize investigative procedures in a genus with limited access to type material and a lack of comprehensive sampling across its total distribution. Our results highlighted lineages that likely represent new species and identified localities that may harbor an as-yet undescribed cryptic species diversity pending further field and laboratory work.

A molecular phylogeny of the sphaerodactylid geckos of the genus Pristurus is inferred based on an alignment of 1845 base pairs (bp) of concatenated mitochondrial (12S) and nuclear (acm4, cmos, rag1 and rag2) genes for 80 individuals, representing 18 of the 23-26 species, and the three subspecies of P. rupestris. The results indicate that P. rupestris is poly-phyletic and includes two highly divergent clades: the eastern clade, found in coastal Iran and throughout the Hajar Moun-tain range in northern Oman and eastern UAE; and the western clade, distributed from central coastal Oman, through Yemen, Saudi Arabia and north to southern Jordan. Inferred haplotype networks for the four nuclear genes show that the eastern and western clades of "P. rupestris" are highly differentiated and do not share any alleles. Moreover, although the two clades are differentiated by a morphological multivariate analysis, no one character or set of characters was found to be diagnostic. Based on the molecular analysis of specimens from the type locality of P. rupestris rupestris, the name P. rupestris is applied to the eastern clade. The name that should apply to the western clade cannot be clarified until morpho-logical and genetic data for >P. rupestris> is available from the vicinity of Bosaso, Somalia, and therefore we refer to it as Pristurus sp. 1. The phylogenetic tree of Pristurus supports the hypothesis that P. celerrimus is sister to all the other spe-cies in the analyses and that the Socotra Archipelago was independently colonized a minimum of two times. Copyright © 2014 Magnolia Press.

Two morphotypes, with a large and small body size, of a brown frog Rana t. tagoi occur sympatrically in the Kinki region, central Honshu of Japan. Previous mitochondrial (mt) DNA genealogical study recognized two main lineages (A and B) and several sublineages in R. tagoi, where the small type was placed in the group A-1b, and the large type in groups A-1a and B-2a. Using haplotype network and structure analysis of three nuclear genes, we examined the discrepancy between morphology and mitochondrial genealogy. The results showed that the small type is reproductively isolated from its co-occurring large type (A-1a or B-2a), and that unlimited gene flow occurred between parapatrically occurring two mtDNA lineages of large types (A-1a and B-2a). Discordant genetic relationships between mtDNA and nuclear DNA results may be caused by the past mitochondrial introgression, and possibly, the incomplete lineage sorting. These results also suggest a heterospecific relationship between the large (A-1a and B-2a) and small types (A-1b). The large type is identified as Rana t. tagoi as it is genetically very close to the topotypes of the nominal subspecies, while the small type remains unnamed.

The Common toad Bufo bufo sensu lato is a widespread, morphologically conserved taxon. Recent studies have uncovered deep genetic differentiation between population groups, highlighting the need to revise the current taxonomy of the group and recognize additional species. Here we investigate patterns of variation in molecular (a mitochondrial DNA restriction enzyme assay and sequence data for two nuclear DNA fragments totalling 979 bp) and 17 morphological variables in Northern France where two of these groups meet (B. bufo sensu stricto and B. spinosus), in order to delineate their contact zone and uncover characters that would allow discrimination of the two taxa. Mitochondrial DNA data show an abrupt transition from areas where B. bufo is present to those inhabited by B. spinosus, with a narrow area of overlap east of the city of Caen. Morphometric characters, particularly those related to the positioning of the parotoid glands and metatarsal tubercle shape and size, proved useful in discriminating between species (AUC ≥ 0.97, kappa ≥ 0.79). We then used the differentiating character states to allocate over 300 museum specimens from Western Europe to either species with consistent results, including comparable values of AUC and kappa of the identification models, indicating that models could successfully be applied across datasets. We summarize available evidence relevant to the delineation of the distribution of B. bufo and B. spinosus in France and discuss the characters differentiating both species in an evolutionary context. In view of the observed morphological and genetic differentiation and the absence of unequivocal evidence for widespread hybridization we support the view that B. bufo and B. spinosus are best considered different species. Finally, we propose that ‘parotoids in parallel position’ and a thin and smooth skin are derived character states for B. bufo over the northern part of its range.

The Common toad Bufo bufo sensu lato is a widespread, morphologically conserved taxon. Recent studies have uncovered deep genetic differentiation between population groups, highlighting the need to revise the current taxonomy of the group and recognize additional species. Here we investigate patterns of variation in molecular (a mitochondrial DNA restriction enzyme assay and sequence data for two nuclear DNA fragments totalling 979 bp) and 17 morphological variables in Northern France where two of these groups meet (B. bufo sensu stricto and B. spinosus), in order to delineate their contact zone and uncover characters that would allow discrimination of the two taxa. Mitochondrial DNA data show an abrupt transition from areas where B. bufo is present to those inhabited by B. spinosus, with a narrow area of overlap east of the city of Caen. Morphometric characters, particularly those related to the positioning of the parotoid glands and metatarsal tubercle shape and size, proved useful in discriminating between species (AUC ≥ 0.97, kappa ≥ 0.79). We then used the differentiating character states to allocate over 300 museum specimens from Western Europe to either species with consistent results, including comparable values of AUC and kappa of the identification models, indicating that models could successfully be applied across datasets. We summarize available evidence relevant to the delineation of the distribution of B. bufo and B. spinosus in France and discuss the characters differentiating both species in an evolutionary context. In view of the observed morphological and genetic differentiation and the absence of unequivocal evidence for widespread hybridization we support the view that B. bufo and B. spinosus are best considered different species. Finally, we propose that ‘parotoids in parallel position’ and a thin and smooth skin are derived character states for B. bufo over the northern part of its range.

An important step in conservation is to identify whether threatened populations are evolutionarily discrete and significant to the species. A prior mitochondrial DNA (mtDNA) phylogeographic study of the California Gnatcatcher (Polioptila californica) revealed no geographic structure and, thus, did not support the subspecies validity of the threatened coastal California Gnatcatcher (P. c. californica). The U.S. Fish and Wildlife Service concluded that mtDNA data alone were insufficient to test subspecies taxonomy. We sequenced eight nuclear loci to search for historically discrete groupings that might have been missed by the mtDNA study (which we confirmed with new ND2 sequences). Phylogenetic analyses of the nuclear loci revealed no historically significant groupings and a low level of divergence (GST = 0.013). Sequence data suggested an older population increase in southern populations, consistent with niche modeling that suggested a northward range expansion following the Last Glacial Maximum (LGM). The signal of population increase was most evident in the mtDNA data, revealing the importance of including loci with short coalescence times. The threatened subspecies inhabits the distinctive Coastal Sage Scrub ecosystem, which might indicate ecological differentiation, but a test of niche divergence was insignificant. The best available genetic, morphological, and ecological data indicate a southward population displacement during the LGM followed by northward range expansion, without the occurrence of significant isolating barriers having led to the existence of evolutionarily discrete subspecies or distinct population segments that would qualify as listable units under the Endangered Species Act.

A new benthic dinoflagellate species, Prorocentrum bimaculatum sp. nov., is studied from Kuwaits marine sediments, based on detailed morphological and molecular data. Cells are large, oblong oval in shape. They are 49.955.3 mu m long and 38.443.2 mu m wide. The ornamentation of this new species is peculiar, and characterized by smooth valves with large pores (0.320.50 mu m) scattered on their surface, except in two circular patches of similar to 15 mu m in diameter, devoid of ornamentation and located on both sides of the valve centers. The periflagellar area is widely triangular, located in a moderate excavation of the right valve, and comprises nine platelets. The intercalary band of P. bimaculatum is smooth. The molecular phylogenetic position of this new taxon was inferred from SSU and LSU rDNA genes. In both phylogenetic analyses, P. bimaculatum branched with high support with Prorocentrum consutum and formed a clade sister to the one including P. lima and related species such as P. arenarium, P. belizeanum, P. hoffmannianum, and P. maculosum. From the phylogenetic study, since most species related to P. bimaculatum are known for their toxic effects and production of okadaic acid, this new species can be considered as a potential toxin producer, but this has to be analyzed.

Little is known about Ziphiidae, elusive deep-diving beaked whales that spend little time at the surface and occur exclusively in deep waters off the continental shelf. Mass strandings of Blainville’s (Mesoplodon densirostris) and Cuvier’s (Ziphius cavirostris) beaked whales have been correlated with sonar exercises conducted by the United States Navy at their sonar-testing facilities in the northern Bahamas. It is not known how naval sonar may be affecting local populations of M. densirostris and Z. cavirostris that inhabit the waters surrounding the testing site. Current regulations and conservation measures established to protect the whales are limited due to an overall lack of understanding in the distribution, abundance, and population structure of beaked whales in the region and globally. To better understand the population structure of these animals, full mitochondrial genomes of both species were generated from 41 stranded and live animals across global ocean basins to analyze the mitogenomic diversity and phylogeographic patterns both around the Bahamas and globally. Within M. densirostris¸ haplotypes were found to be divided into two distinct geographic clades representing possible Atlantic and Pacific sub species or species. In Z. cavirostris, 3 major clades were discovered, however, in two clades both Atlantic and Pacific animals were found, indicating that inter-ocean migration events have occurred relatively recently, or that there is ongoing gene flow for this species between ocean basins. Further sampling of both species and additional data from nuclear DNA will be required to fully resolve this and to better understand and mitigate the effects of naval activities on beaked whale populations within the northern Bahamas and elsewhere.