An integrative multidisciplinary approach to understanding cryptic divergence in Brazilian species of the Anastrepha fraterculus complex (Diptera: Tephritidae)

An integrative multidisciplinary approach was used to delimit boundaries among cryptic species within the Anastrepha fraterculus complex in Brazil. Sexual compatibility, courtship and sexual acoustic behaviour, female morphometric variability, variation for the mitochondrial gene COI, and the presence of Wolbachia were compared among A. fraterculus populations from the Southern (Vacaria, Pelotas, Bento Gonçalves, São Joaquim) and Southeastern (Piracicaba) regions of Brazil. Our results suggest full mating compatibility among A. fraterculus populations from the Southern region and partial pre‐zygotic reproductive isolation of these populations when compared with the population from the Southeastern region. A. fraterculus populations from both regions differed in the frequency of courtship displays and aspects of the calling phase and mounting acoustic signal. Morphometric analysis showed differences between Southern region and Southeastern region samples. All populations analyzed were infected with Wolbachia. The trees generated from the COI sequencing data are broadly congruent with the behavioural and morphometric data with the exception of one Southern population. The likely mechanisms by which A. fraterculus populations might have diverged are discussed in detail based on behavioural, morphometric, molecular genetics, and biogeographical studies.     

An integrative taxonomic review of the Natal mountain catfish,Amphilius natalensis Boulenger 1917 (Siluriformes,Amphiliidae), with description of four new species

Species delimitation is a permanent issue in systematics. The increasing recognition of geographically isolated populations as independent lineages allowed by new methods of analysis has inflated the species-populations dilemma, which involves deciding whether to consider separate lineages as different species or structured genetic populations. This is commonly observed between fishes of adjacent river basins, with some lineages being considered allopatric sister species and others considered isolated populations or variants of the same species. Pseudocorynopoma doriae is a characid diagnosed from its single congener by the number of anal-fin rays and sexually dimorphic characters of males, including distinct fin colouration. The authors found variation in the colour pattern between isolated populations previously identified as P. doriae but no variation in scale or fin-ray counts. They analysed molecular evidence at the population level and morphological differences related to life history (e.g., colour dimorphism related to inseminating behaviour). The results provide compelling evidence for the recognition of a new species of Pseudocorynopoma despite the lack of discrete differences in meristic data. The recognition of the new species is consistent with biogeographical evidence for the long-term isolation of the respective river drainages and with differences between the ichthyofaunal communities of these rivers.     

Untangling the Spurilla neapolitana (Delle Chiaje, 1841) species complex: a review of the genus Spurilla Bergh, 1864 (Mollusca: Nudibranchia: Aeolidiidae)

Spurilla neapolitana (Delle Chiaje, 1823) was considered to be a species with a broad geographic range and substantial colour variability; however, analyses of mitochondrial and nuclear gene data revealed that it is a complex of five distinct species. Further anatomical and morphological examinations determined that coloration is one of the main diagnostic traits for all five species, although some display substantial colour pattern variation. As a result of this study, S. neapolitana is determined to be restricted to the Mediterranean and eastern Atlantic. Spurilla sargassicola Bergh, 1871 from the Caribbean is redescribed and confirmed as a valid species. The name Spurilla braziliana MacFarland, 1909 is retained for western Atlantic and Pacific populations. Two new species are described herein. S purilla onubensis sp. nov. occurs in Europe, with a range overlapping that of S. neapolitana. Finally, S purilla dupontae sp. nov. is found in the Bahamas. © 2014 The Linnean Society of London     

Taking the discovery approach in integrative taxonomy: decrypting a complex of narrow‐endemic Alpine harvestmen (Opiliones: Phalangiidae: Megabunus)

Species delimitation is fundamental for biological studies, yet precise delimitation is not an easy task, and every involved approach has an inherent failure rate. Integrative taxonomy, a method that merges multiple lines of evidence, can profoundly contribute to reliable alpha‐taxonomy and shed light on the processes behind speciation. In this study, we explored and validated species limits in a group of closely related Megabunus harvestmen (Eupnoi, Phalangiidae) endemic to the European Alps. Without a priori species hypotheses, we used multiple sources of inference, including mitochondrial and multilocus nuclear DNA, morphometrics and chemistry. The results of these discovery approaches revealed morphological crypsis and multiple new species within two of the five hitherto known species. Based on our analyses, we discussed the most plausible evolutionary scenarios, invoked the most reasonable species hypotheses and validated the new species limits. Building upon the achieved rigour, three new species, Megabunus cryptobergomas Muster and Wachter sp. nov., Megabunus coelodonta Muster and Steiner sp. nov., and Megabunus lentipes Muster and Komposch sp. nov., are formally described. In addition, we provide a dichotomous morphological key to the Megabunus species of the Alps. Our work demonstrates the suitability of integrative, discovery‐based approaches in combination with validation approaches to precisely characterize species and enabled us to implement nomenclatural consequences for this genus.     

Assessing the diversity and distribution of Cephalothrix species (Nemertea: Palaeonemertea) in European waters by comparing different species delimitation methods

Soft‐bodied marine taxa, like ribbon worms (Nemertea), often lack clear diagnostic morphological characters impeding traditional species delimitation. Therefore, recent studies concentrated on molecular genetic methods to solve taxonomic issues. Different delimitation methods were employed to explore species boundaries and the presence of cryptic species. However, the performance of the different delimitation methods needs to be tested. A particularly promising nemertean genus in this regard is the palaeonemertean genus Cephalothrix that is commonly found in European waters. In order to gain information on the number and distribution of European cephalotrichids and to test different tree‐based and non‐tree‐based delimitation methods, we analyzed a dataset comprising the barcoding region of the mitochondrial cytochrome c oxidase subunit I (COI) of 215 European Cephalothrix specimens, of which 78 were collected for this study. Our results show the presence of 12–13 European lineages of which several can be assigned to known European species. Analyzing a second dataset comprising 74 additional sequences from the Pacific and the Atlantic Oceans helped identify some of the unassigned European specimens. One resulting clade seems to represent a non‐native introduced Cephalothrix species, while another has never been recorded from Europe before. In our analysis, especially the tree‐based methods and the phylogenetic analysis proved to be a useful tool when delimiting species. It remains unclear whether the different identified clades result from cryptic speciation or from a high genetic variability of the COI gene.     

Phylogenetics and biogeography of the two‐wing flyingfish (Exocoetidae: Exocoetus)

Two‐wing flyingfish (Exocoetus spp.) are widely distributed, epipelagic, mid‐trophic organisms that feed on zooplankton and are preyed upon by numerous predators (e.g., tunas, dolphinfish, tropical seabirds), yet an understanding of their speciation and systematics is lacking. As a model of epipelagic fish speciation and to investigate mechanisms that increase biodiversity, we studied the phylogeny and biogeography of Exocoetus, a highly abundant holoepipelagic fish taxon of the tropical open ocean. Morphological and molecular data were used to evaluate the phylogenetic relationships, species boundaries, and biogeographic patterns of the five putative Exocoetus species. We show that the most widespread species (E. volitans) is sister to all other species, and we find no evidence for cryptic species in this taxon. Sister relationship between E. monocirrhus (Indo‐Pacific) and E. obtusirostris (Atlantic) indicates the Isthmus of Panama and/or Benguela Barrier may have played a role in their divergence via allopatric speciation. The sister species E. peruvianus and E. gibbosus are found in different regions of the Pacific Ocean; however, our molecular results do not show a clear distinction between these species, indicating recent divergence or ongoing gene flow. Overall, our phylogeny reveals that the most spatially restricted species are more recently derived, suggesting that allopatric barriers may drive speciation, but subsequent dispersal and range expansion may affect the distributions of species.     

Molecular phylogeography of the microturbellarian Monocelis lineata (Platyhelminthes: Proseriata) in the North‐East Atlantic

Monocelis lineata is a complex of cryptic species (three in the Mediterranean and one in the Atlantic) widespread in midlittoral habitats. Throughout the range, populations with or without an ocular pigmented shield are found. We investigated the genetic structure of the North‐East Atlantic populations with the aim of shedding light on their phylogeography and reconstructing possible patterns of recolonization after the Würmian glaciation. Fourteen samples were investigated using cytochrome c oxidase subunit I (COI) and 13 by inter‐simple sequence repeats (ISSRs). COI did not exhibit a clear pattern of decreased genetic diversity along a latitudinal gradient. Populations from Ferrol (Spain), Doolin (Ireland), and Helsingør (Denmark) showed a higher genetic variability, whereas a reduction in the number of haplotypes was found at the northernmost edge of the distribution and in northern Ireland and Scotland. Two genetically differentiated areas (southern Europe and south‐western Ireland versus northern Atlantic) were revealed by ISSR data. The results obtained provided evidence of three refugia (Iberian Peninsula, south‐western Ireland, and North Sea), and the occurrence of secondary contacts that shaped the genetic variability of some of the populations examined. Two different recolonization pathways of north‐western Europe during the post‐Würmian glaciations have been detected. Furthermore, ISSR analysis provided evidence of genetic divergence among populations with and without pigmented eyespot, suggesting the action of ecological differentiation. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 117–135.     

A molecular and morphological approach on the taxonomic status of the Brazilian species of Palaemon (Decapoda,Palaemonidae)

Previous studies have indicated the occurrence of nine species of Palaemon in Brazil. However, the extensive variability in their diagnostic characters and the geographic distribution of some species raises doubts about the validity of some taxa, potentially indicating the occurrence of species not yet reported from Brazil or even not described. To solve taxonomic issues with Palaemon species in Brazil, we carried out a multigene analysis by using partial fragments of 16S and COI mitochondrial genes and the nuclear H3. Phylogenetic relationships were inferred by maximum likelihood and Bayesian inference. The external morphology corresponding to the terminal clades obtained in the molecular phylogram was analysed to identify diagnostic characters that allow the recognition of the groups and the correct taxa assignment. The combined analysis of molecular and morphological data confirms the existence of nine species of Palaemon in Brazil, five from the “Palaemon” lineage (P. mercedae and four Palaemon stensu stricto—P. argentinus, P. octaviae—first record in Brazil, P. northropi and P. paivai) and four of the “Alaocaris” lineage (P. carteri, P. ivonicus, P. pandaliformis and P. yuna). The possibility of further cryptic speciation is also pointed out. In addition to confirming the validity of the nine species, the results indicate the need for taxonomic adjustments and corresponding changes in the geographic distribution of Palaemon species occurring in Brazil.     

Host‐associated genetic differentiation in a seed parasitic weevil Rhinusa antirrhini (Coleptera: Curculionidae) revealed by mitochondrial and nuclear sequence data

Plant feeding insects and the plants they feed upon represent an ecological association that is thought to be a key factor for the diversification of many plant feeding insects, through differential adaptation to different plant selective pressures. While a number of studies have investigated diversification of plant feeding insects above the species level, relatively less attention has been given to patterns of diversification within species, particularly those that also require plants for oviposition and subsequent larval development. In the case of plant feeding insects that also require plant tissues for the completion of their reproductive cycle through larval development, the divergent selective pressure not only acts on adults, but on the full life history of the insect. Here we focus attention on Rhinusa antirrhini (Curculionidae), a species of weevil broadly distributed across Europe that both feeds on, and oviposits and develops within, species of the plant genus Linaria (Plantaginaceae). Using a combination of mtDNA (COII) and nuclear DNA (EF1‐α) sequencing and copulation experiments we assess evidence for host associated genetic differentiation within R. antirrhini. We find substantial genetic variation within this species that is best explained by ecological specialisation on different host plant taxa. This genetic differentiation is most pronounced in the mtDNA marker, with patterns of genetic variation at the nuclear marker suggesting incomplete lineage sorting and/or gene flow between different host plant forms of R. antirrhini, whose origin is estimated to date to the mid‐Pliocene (3.77 Mya; 2.91–4.80 Mya).     

Barcoding of Central European Cryptops centipedes reveals large interspecific distances with ghost lineages and new species records from Germany and Austria (Chilopoda,Scolopendromorpha)

In order to evaluate the diversity of Central European Myriapoda species in the course of the German Barcode of Life project, 61 cytochrome c oxidase I sequences of the genus Cryptops Leach, 1815, a centipede genus of the order Scolopendromorpha, were successfully sequenced and analyzed. One sequence of Scolopendracingulata Latreille, 1829 and one of Theatopserythrocephalus Koch, 1847 were utilized as outgroups. Instead of the expected three species (Cryptopsparisi Brolemann, 1920; Cryptopsanomalans Newport, 1844; Cryptopshortensis (Donovan, 1810)), analyzed samples included eight to ten species. Of the eight clearly distinguishable morphospecies of Cryptops, five (Cryptopsparisi; Cryptopscroaticus Verhoeff, 1931; Cryptopsanomalans; Cryptopsumbricus Verhoeff, 1931; Cryptopshortensis) could be tentatively determined to species level, while a further three remain undetermined (one each from Germany, Austria and Croatia, and Slovenia). Cryptopscroaticus is recorded for the first time from Austria. A single specimen (previously suspected as being Cryptopsanomalans), was redetermined as Cryptopsumbricus Verhoeff, 1931, a first record for Germany. All analyzed Cryptops species are monophyletic and show large genetic distances from one another (p-distances of 13.7–22.2%). Clear barcoding gaps are present in lineages represented by >10 specimens, highlighting the usefulness of the barcoding method for evaluating species diversity in centipedes. German specimens formally assigned to Cryptopsparisi are divided into three clades differing by 8.4–11.3% from one another; their intra-lineage genetic distance is much lower at 0–1.1%. The three clades are geographically separate, indicating that they might represent distinct species. Aside from Cryptopsparisi, intraspecific distances of Cryptops spp. in Central Europe are low (<3.3%).     

Three new species of Stiphrornis (Aves: Muscicapidae) from the Afro-tropics,with a molecular phylogenetic assessment of the genus

We describe three new species of forest robin in the genus Stiphrornis; two from West Africa and one from the Congo Basin. Each species represents a distinct phylogenetic lineage based on genetic analysis. In addition to genetic differentiation, each new species is diagnosable from other Stiphrornis lineages by morphology, and by plumage. One of the new species appears to be restricted to the Central and Brong-Ahafo Regions of Ghana, and another is restricted to Benin and the Central Region of Ghana. In Ghana, these two new species presumably come into contact with Stiphrornis erythrothorax (Western Region of Ghana and westward), and there is evidence that one of the new species has a distinguishably different song from erythrothorax. The distribution of the third new species is primarily on the south bank of the Congo River, near the city of Kisangani. Recognition of these species provides additional evidence that Afrotropical forests are harbouring substantial cryptic diversity, and that our knowledge of the drivers of this diversity remains poorly documented across the region.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:BF2A0BE6-1140-4EFF-9035-380D61AB03AE     

Molecular data illuminate cryptic nudibranch species: the evolution of the Scyllaeidae (Nudibranchia: Dendronotina) with a revision of Notobryon

Scyllaeidae represents a small clade of dendronotoid nudibranchs. Notobryon wardi Odhner, 1936, has been reported to occur in tropical oceans from the Indo‐Pacific and eastern Pacific to temperate South Africa. The systematics of Notobryon has not been reviewed using modern systematic tools. Here, specimens of Notobryon were examined from the eastern Pacific, the Indo‐Pacific, and from temperate South Africa. Additionally, representatives of Scyllaea and Crosslandia were studied. Scyllaeidae was found to be monophyletic. Notobryon was also found to be monophyletic and is the sister group to Crosslandia plus Scyllaea. The molecular data also clearly indicate that within Notobryon, at least three distinct species are present, two of which are here described. Genetic distance data indicate that eastern Pacific and South African exemplars are 10–23% divergent from Indo‐Pacific exemplars of Notobryon wardi. Scyllaea pelagica has been regarded as a single, circumtropical species. Our molecular studies clearly indicate that the Atlantic and Indo‐Pacific populations are distinct and we resurrect Scyllaea fulva Quoy & Gaimard, 1824 for the Indo‐Pacific species. Our morphological studies clearly corroborate our molecular findings and differences in morphology distinguish closely related species. Different species clearly have distinct penial morphology. These studies clearly reinforce the view that eastern Pacific, Indo‐Pacific, and temperate biotas consist largely of distinct faunas, with only a minor degree of faunal overlap. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 165 , 311–336.     

Taxonomy and phylogeny of Lineus torquatus and allies (Nemertea,Lineidae) with descriptions of a new genus and a new cryptic species

The heteronemertean genus Lineus Sowerby, 1806 has been badly in need of revision because of its apparent non-monophyly. In this paper, we focus on Lineus torquatus Coe, 1901, one of the heteronemertean species that occur commonly in waters around the North Pacific, as well as a few other allied species distributed in the western North Pacific, including Lineus alborostratus Takakura, 1898 and Cerebratulus montgomeryi Coe, 1901. Based on phylogenetic analyses using 16S, COI, 28S, 18S, and H3 gene and ITS sequences, we detected a well-supported clade comprised of heteronemerteans with a frontal white band on the head, to which we add Kulikovia gen. nov. This genus is nested within a more comprehensive, highly supported clade, here named the Siphonenteron-clade, which contains Tenuilineus bicolour (Verrill, 1892), Lineus flavescens Coe, 1905, Siphonenteron bilineatum (Renier, 1804), S. cf. bilineatum, Lineus cf. caputornatus, and Lineus sp. from Guam. Our analyses confirmed the presence of a cryptic species of what was formerly known as the cherry-red and reddish forms of Lineus torquatus, herein described as Kulikovia manchenkoi sp. nov. based on some external characters, internal morphology, and the four genetic markers (COI, 16S, H3, and ITS). In contrast to the species pair K. torquata–K. manchenkoi, the reddish form of K. alborostrata does not differ genetically from the typical form of this species. The significance of the external and internal characters for distinguishing cryptic species is discussed.

http://zoobank.org/urn:lsid:zoobank.org:pub:9BECBC60-9C82-48EC-AD36-FC564D82A5BChttp://zoobank.org/urn:lsid:zoobank.org:act:D02B2339-4F65-4517-9B13-DD4AAB0C55C5http://zoobank.org/urn:lsid:zoobank.org:act:8D239B62-E655-4721-90F0-A4944DD8A3C7     

An integrative anatomical,morphological, micromorphological and molecular approach to Turkish epidendroid and orchidoid species (Orchidaceae)

We aimed to describe and analyse the morphological, anatomical and micromorphological traits of 36 Turkish orchids representing 12 genera (e.g. Anacamptis, Cephalanthera, Dactylorhiza, Orchis, Serapias) in detail and analyse their usability for solving phylogenetic and taxonomic issues. We applied UPGMA cluster analysis to anatomical, morphological and micromorphological characters such as root tubers, leaf and flower structures, pit, sclerenchymatic sheath, vascular bundle shape, crystal and starch, exodermis and endodermis structure, stomata type, bulliform cells in roots, shoots and leaves, surface structures like papillae, hairs and ornamentation on flower parts, leaves, fruits and seeds. Furthermore, in a phylogenetic framework, we analysed nuclear ribosomal ITS diversity in the same orchid species belonging, and in a combined Bayesian phylogenetic analysis based on anatomical, morphological, micromorphogical and ITS data we confirmed the usefulness of multiple data sets for effectively assessing taxonomically critical orchids. In the combined analysis, all genera were resolved as monophyletic with topologies congruent with recently published more thorough molecular phylogenetic reconstructions, while the trees obtained by seprately analysing the ITS and the anatomical, morphological, micromorphogical data were less resolved and partly inconclusive.