Complete mitochondrial genome sequence and phylogenetic position of the Amu Darya sturgeon,Pseudoscaphirhynchus kaufmanni (Acipenseriformes: Acipenseridae)

In this study, we successfully assembled the complete mitochondrial genome of the Amu Darya sturgeon Pseudoscaphirhynchus kaufmanni. Based on this mitochondrial genome and previously published mitochondrial genomes of members of the Acipenseridae family, we assessed the phylogenetic position of P. kaufmanni using maximum likelihood and Bayesian inference for phylogeny reconstruction. The resultant phylogenetic trees were well-resolved, with congruence between different phylogenetic methods. This robust phylogenetic analysis elucidated the relationship among the four acipenserid genera and strongly supported the division of the family into three main clades. Evaluation of molecular phylogeny using maximum likelihood and Bayesian analysis led to the following conclusions: (a) the most basal position within the Acipenseridae remains in the clade containing Acipenser oxyrinchus and Acipenser sturio; (b) the genus Scaphirhynchus belongs to the Atlantic clade and is a sister group of the remaining species of the clade; and (c) the close relationship between P. kaufmanni and Acipenser stellatus is well supported.     

Phylogenetic analysis of the family Ariidae (Ostariophysi: Siluriformes), with a hypothesis on the monophyly and relationships of the genera

Ariid monophyly and intrafamilial relationships are investigated based on cladistic analysis of 230 morphological characters. Terminal taxa examined include whenever possible type‐species, or the most morphologically similar species to the type‐species of the nominal genera, and the largest possible number of species, including cleared and stained specimens, available in zoological collections. Previous hypotheses about monophyly of the Ariidae are strongly corroborated by new synapomorphies discovered in the present study. The subfamily Galeichthyinae and the remaining ariids are strongly supported by new morphological characters. The monotypic subfamily Bagreinae is recognized as the sister group to all nongaleichthyin ariids, supported by a large series of exclusive synapomorphies. A new concept of Ariinae is presented: the subfamily is found to be unequivocally monophyletic and includes all ariid genera, except Galeichthys and Bagre. New data supporting the monophyly of the genera included in the Ariinae are introduced and previous hypotheses of monophyly, species composition, morphological definition, and relationships are reviewed and discussed.     

Cranial morphology of the Stellate Sturgeon,Acipenser stellatus Pallas 1771 (Acipenseriformes,Acipenseridae), with notes on the skulls of other sturgeons

Extant members of Acipenseridae are generally classified in four genera: Scaphirhynchus, Pseudoscaphirhynchus, Huso and “Acipenser,” which is widely recognized to be paraphyletic. Advances have been made in understanding the systematic relationships among sturgeons based on both morphological and molecular data. Analysis of mitochondrial DNA data suggested that Pseudoscaphirhynchus should be regarded as nested within “Acipenser,” specifically as sister group to the Stellate Sturgeon, A. stellatus. Recent morphological analyses also recovered this relationship, supported by a number of osteological synapomorphies, although these results were based on few and relatively small individuals. Here we describe the anatomy of the skull of A. stellatus based on newly prepared specimens of adult individuals, as well as examination of a large number of preserved individuals representing a broad range of ontogenetic stages. We present new anatomical data from all regions of the skull (dermatocranium, neurocranium, viscerocranium) and offer interpretations of these and other characters. In particular, we describe the allometry in the snout of A. stellatus, which undergoes substantial elongation relative to other sturgeons. Aspects of the skull of A. stellatus are compared to other members of the family, specifically the course of the occipital sensory canal and the morphology and distribution of cranial spines.     

Genetic distinction of pallid, shovelnose, and Alabama sturgeon: emerging species and the US Endangered Species Act

The sturgeon genus Scaphirhynchus consists of threerecognized species. Pallid and shovelnose sturgeon (S. albusand S. platorynchus, respectively) are sympatric in theMissouri and lower Mississippi Rivers of the central United States. TheAlabama sturgeon (S. suttkusi) is endemic to the nearby MobileRiver drainage and is isolated geographically from the other twospecies. Pallid sturgeon and the extremely rare Alabama sturgeon arelisted as endangered under the US Endangered Species Act (ESA).In contrast, shovelnose sturgeon are relatively common and are notlisted. Despite these taxonomies and morphological evidence, somebiologists have questioned the genetic and taxonomic distinctions of thethree species, thus raising doubts concerning the validity of protectingpallid and Alabama sturgeon under the ESA. To investigate thesequestions, we compared a 436 base-pair sequence of the mitochondrial DNA(mtDNA) control region among the three species. We observed 16 mtDNAhaplotypes defined by 27 single base-pair substitutions (transitions)and one single base-pair insertion/deletion (indel) among 78individuals examined. The maximum sequence divergence among thosehaplotypes (2.06%) was less than values usually observed betweenfish species. However, Alabama sturgeon (n = 3) weredistinguished from the other two taxa (n = 75) by aunique base-pair substitution and haplotype, and pallid and shovelnosesturgeon at their northern range of natural sympatry (upper MissouriRiver) did not share any haplotypes. On the other hand, only frequencydifferences among shared haplotypes distinguished (P < 0.01)pallid and shovelnose sturgeon at their southern range of naturalsympatry (Atchafalaya River), and genetic distances between northern andsouthern localities for each species were nearly as large as thedistances between species. These latter results are consistent withseveral hypotheses, including reports (based on morphology) of putativenatural hybrids in the Atchafalaya River but not in the upper MissouriRiver. Overall, these mtDNA results indicate significant reproductiveisolation between pallid and shovelnose sturgeon in areas of naturalsympatry, and recent evolutionary divergence of Alabama sturgeon. ThesemtDNA results provide the first molecular genetic evidence fordistinguishing the three Scaphirhynchus species and, coupledwith morphological and biogeographic data, indicate that pallid andAlabama sturgeon should be evaluated as distinct species under theESA.     

Microsatellite variation among River Sturgeons of the genus Scaphirhynchus (Actinopterygii: Acipenseridae): a preliminary assessment of hybridization*

Microsatellite variation from 13 disomic loci is reported for a total of 208 individuals of the genus Scaphirhynchus. This includes 105 individuals of the pallid sturgeon (Scaphirhynchus albus) from the lower Mississippi River, 11 pallid sturgeon from the Upper Mississippi and Missouri rivers, 65 shovelnose sturgeon (S. platorynchus) from the lower Mississippi River, six Alabama sturgeon (S. suttkusi), and 21 individuals of sturgeon identified as intermediate between S. albus and S. platorynchus. Results indicate that all five of the above population/species units are significantly differentiated from one another based on pairwise F_ST estimates. Locus Spl‐7 was diagnostic for the Alabama sturgeon and serves to further differentiate this allopatric species from other Scaphirhynchus. Classification of genotypes with and without a priori designations failed to clearly delineate the species and intermediates in the latter case but was successful for the species but not in the intermediates in the former case. The presence of six unique alleles in five of the 21 morphologically ‘intermediate’ sturgeon examined requires additional evaluation but suggests that these individuals are possibly not the result of hybridization. We hope that raising these important issues will bring all stakeholders to the table to establish a concerted effort needed for both morphological and molecular analyses to adequately address the question of hybridization and the origin of the morphological variation in these fishes.     

The subfamily Myoxocephalinae of cottid fishes (Cottidae): Genetic divergence and phylogenetic relationships

A comprehensive study based on molecular marker and karyotype analyses has provided evidence for the monophyly of the subfamily Myoxocephalinae, which includes the genera Myoxocephalus, Megalocottus, Microcottus, Porocottus, Enophrys and Argyrocottus. In addition, the karyotype of the threadfoot sculpin Argyrocottus zanderi Herzenstein 1892 has been studied for the first time. Marker traits of karyotypes identified 13 species among six cottid genera. As the molecular genetic results confirmed, the subfamily is divided into two groups corresponding to Enophrys and Myoxocephalus. The molecular genetic data did not support the formation of tribes within the subfamily Myoxocephalinae, as proposed earlier based on morphological characters. Moreover, the genera Trichocottus and Taurocottus should be excluded from the Myoxocephalinae. The evolutionary transformations of karyotypes in cottid fish tended towards a reduction in the number of chromosomes and chromosome arms.     

Comparative neuromorphology of the telencephalon of sturgeon of the genera Acipenser, Huso and Scaphirhynchus (Actinopterygii; Acipenseridae)

Brain anatomy and structure, composition of the neurons, and cytoarchitectural maps were compared between five species of Acipenser, one species of Huso and two species of Scaphirhynchus. Morphological differences in the forebrain structure between species and genera were apparent. The evolutionary history of the Acipenseridae has been marked by an extensive mosaic radiation in the evolutionary development of the inner structure of their brains. Members of Scaphirhynchus have a very simple telencephalon structure differing significantly from that of Huso and Acipenser. It is clear that neuromorphological variation of sturgeon species will serve as an additional data useful in understanding the evolutionary processes and history associated with the speciation in Acipenseridae.     

Phylogenetic relationships,subdivision, and biogeography of the cyprinid tribe Labeonini (sensu ) (Teleostei: Cypriniformes), with comments on the implications of lips and associated structures in the labeonin classification

The Labeonini (sensu Rainboth, 1991) is a tribe of the subfamily Cyprininae, the largest subfamily of Cypriniformes. With around 400 species in 34 genera, this tribe is widely distributed in the freshwaters of tropical Africa and Asia. Most species are adapted to fast-flowing streams and rivers, and exhibit unique morphological modifications associated with their lips and other structures around the mouth. The monophyly of this tribe has been tested and generally accepted in previous morphological and molecular studies. The major objectives of this study were to reconstruct the phylogenetic relationships within the tribe Labeonini, test its monophyly and explore the taxonomic subdivisions, intrarelationships and biogeography of the group. The value of the morphological characters associated with the lips and other associated structures in the taxonomic classification of labeonins was also discussed. Nucleotide sequences (3867 bp) of four unlinked nuclear loci were obtained from 51 species in 18 Labeonini genera from throughout the range of the tribe. Maximum parsimony, partitioned maximum likelihood and partitioned Bayesian analyses were used for phylogenetic inference from combined and separate gene data sets. Based on our results, the monophyly of Labeonini was well supported. Two major clades could be recovered within the tribe. Three subclades could further be recognized from the first clade. These clades/subclades are not consistent with groupings of any of previous workers using either morphological or molecular characters for phylogenetic inference. Only five currently recognized genera in this analysis are monophyletic. The similarity between some lips and associated structures (e.g. suctorial discs) of labeonins may due to convergence or parallelism instead of common ancestry. Labeonins of Southeast Asia, India and China are closely related to each other; the multiple clades of African taxa do not form a single monophyletic group, indicating multiple, independent dispersal events of labeonins into Africa from Asia.     

Morphology agrees with molecular data: phylogenetic affinities of Euptychiina butterflies (Nymphalidae: Satyrinae)

Euptychiina is the most species‐rich subtribe of Neotropical Satyrinae, with over 450 known species in 47 genera (14 monotypic). Here, we use morphological characters to examine the phylogenetic relationships within Euptychiina. Taxonomic sampling included 105 species representing the majority of the genera, as well as five outgroups. A total of 103 characters were obtained: 45 from wing pattern, 48 from genitalia and 10 from wing venation. The data matrix was analysed using maximum parsimony under both equal and extended implied weights. Euptychiina was recovered as monophyletic with ten monophyletic genera, contrasting previous DNA sequence‐based phylogenies that did not recover the monophyly of the group. In agreement with sequence‐based hypotheses, however, three main clades were recognized: the ‘Megisto clade’ with six monophyletic and three polyphyletic genera, the ‘Taygetis clade’ with nine genera of which three were monophyletic, and the ‘Pareuptyhia clade’ with four monophyletic and two polyphyletic genera. This is the first morphology‐based phylogenetic hypothesis for Euptychiina and the results will be used to complement molecular data in a combined analysis and to provide critical synapomorphies for clades and genera in this taxonomically confused group.     

Systematics and evolution of syllids (Annelida,Syllidae)

A large, combined phylogenetic analysis (including morphological and molecular data from 18S rDNA, 16S rDNA and cytochrome c oxidase subunit I), with the highest number of species and genera of Syllidae studied to date (213 terminals), is examined. The data were explored with different parameters and optimality criteria (parsimony, likelihood, and bayesian inference). The monophyly of Syllidae and most of the traditional subfamilies is supported. The subfamily Eusyllinae is polyphyletic, as currently delineated, but it is herein reorganized and its diagnosis modified to be a valid group. Additional well supported clades arise. The phylogenetic relationships of the well known and established genera, as well as several enigmatic genera (e.g. Anguillosyllis, Paraopisthosyllis and Parahaplosyllis), the position of which in syllid taxonomy was uncertain or dubious to date, are clarified. The results corroborate previous hypotheses about the evolution of the reproductive and brooding modes. Within Syllinae, the nature of the stolon is phylogenetically informative. The classification of the whole family is revised and discussed on the basis of this phylogenetic hypothesis. © The Willi Hennig Society 2011.     

Phylogeny of the cricket subfamily Eneopterinae (Orthoptera, Grylloidea, Eneopteridae) based on four molecular loci and morphology

The phylogenetic relationships of 39 species of Eneopterinae crickets are reconstructed using four molecular markers (16S rRNA, 12S rRNA, cytochrome b, 18S rRNA) and a large morphological data set. Phylogenetic analysis via direct optimisation of DNA sequence data using parsimony as optimality criterion is done for six combinations of weighting parameter sets in a sensitivity analysis. The results are discussed in a twofold purpose: first, in term of significance of the molecular markers for phylogeny reconstruction in Ensifera, as our study represents the first molecular phylogeny performed for this insect suborder at this level of diversity; second, in term of corroboration of a previous phylogeny of Eneopterinae, built on morphological data alone. The four molecular markers all convey phylogenetic signal, although variously distributed on the tree. The monophyly of the subfamily, that of three over five tribes, and of 10 over 13 genera, are recovered. Finally, previous hypotheses on the evolution of acoustic devices and signals in the Eneopterinae clade are briefly tested, and supported, by our new data set.     

Mitochondrial and nuclear markers support the monophyly of Dolichopodidae and suggest a rapid origin of the subfamilies (Diptera: Empidoidea)

Abstract. The Dolichopodidae is a species‐rich dipteran group with almost 7000 described species. The monophyly of the subfamilies and their relationships remain largely unknown because the polarities of key morphological characters are unclear and molecular data are available only for 9 of the 19 proposed subfamilies. Here we test whether molecular data from two nuclear (18S, 28S) and four mitochondrial (12S, 16S, Cytb, COI) genes can resolve the higher‐level relationships within the family. Our study is based on 76 Oriental species from 12 dolichopodid subfamilies and uses eight species of Empididae and Hybotidae as outgroups. Parsimony and likelihood analyses confirm the monophyly of the Dolichopodidae, as well as the monophyly of five of the ten subfamilies represented by more than two species [Sympycninae, Sciapodinae, Dolichopodinae, Hydrophorinae (excluding tribe Aphrosylini), Neurigoninae]. There is strong support for restoring the tribe Aphrosylini as a separate subfamily Aphrosylinae. The monophyly of Medeterinae, Peloropeodinae and Diaphorinae is dependent on which tree reconstruction technique is used, how indels are coded, and whether the fast‐evolving sites are excluded. Overall, we find that our sample of Oriental species is largely compatible with the subfamily concepts that were developed for the northern temperate fauna. However, our data provide little support for relationships between the subfamilies. Branch lengths, saturation, and distance plots suggest that this is probably the result of the rapid origin of dolichopodid subfamilies over a relatively short time. We find that genera that are difficult to place into subfamilies based on morphological characters are generally also difficult to place using molecular data. We predict that a dense, balanced taxon sample and protein‐encoding nuclear genes will be needed to resolve the higher‐level relationships in the Dolichopodidae.     

Sound Production in Sturgeon Scaphirhynchus albus and S. platorynchus (Acipenseridae)

Sound production has been recently discovered in several species of Acipenser. Our work has focused on testing for sound production in species of sturgeon in the genus Scaphirhynchus. We discovered that pallid sturgeon Scaphirhynchus albus and shovelnose sturgeon, S. albus produce sounds during the breeding season. These signals may be used as part of efforts to localize populations of sturgeon in the field, including the Alabama sturgeon, S. suttkusi.     

Reconstructing the phylogeny of the Sipuncula

Sipunculans are marine spiralian worms with possible close affinities to the Mollusca or Annelida. Currently 147 species, 17 genera, 6 families, 4 orders and 2 classes are recognized. In this paper we review sipunculan morphology, anatomy, paleontological data and historical affiliations. We have conducted cladistic analyses for two data sets to elucidate the phylogenetic relationships among sipunculan species. We first analyzed the relationships among the 45 species of Phascolosomatidea with representatives of the Sipunculidea as outgroups, using 35 morphological characters. The resulting consensus tree has low resolution and branch support is low for most branches. The second analysis was based on DNA sequence data from two nuclear ribosomal genes (18S rRNA and 28S rRNA) and one nuclear protein-coding gene, histone H3. Outgroups were chosen among representative spiralians. In a third analysis, the molecular data were combined with the morphological data. Data were analyzed using parsimony as the optimality criterion and branch support evaluated with jackknifing and Bremer support values. Branch support for outgroup relationships is low but the monophyly of the Sipuncula is well supported. Within Sipuncula, the monophyly of the two major groups, Phascolosomatidea and Sipunculidea is not confirmed. Of the currently recognized families, only Themistidae appears monophyletic. The Aspidosiphonidae, Phascolosomatidae and Golfingiidae would be monophyletic with some adjustments in their definition. The Sipunculidae is clearly polyphyletic, with Sipunculus nudus as the sister group to the remaining Sipuncula, Siphonosoma cumanense the sister group to a clade containing Siphonosoma vastumand the Phascolosomatidea, and Phascolopsis gouldi grouping within the Golfingiiformes, as suggested previously by some authors. Of the genera with multiple representatives, only Phascolosoma and Themiste are monophyletic as currently defined. We are aiming to expand our current dataset with more species in our molecular database and more detailed morphological studies.     

The ant subfamily Pseudomyrmecinae (Hymenoptera: Formicidae): phylogeny and evolution of big-eyed arboreal ants

Abstract. The ant subfamily Pseudomyrmecinae comprises three genera of hyperoptic, arboreal ants, widely distributed in tropical and subtropical regions: Pseudomyrmex (∼200 species, New World), Myrcidris (two species, South America) and Tetraponera (∼100 species, Palaeotropics). The phylogenetic relationships among these ants were investigated using DNA sequence data (∼5.2 kb from 18S rDNA, 28S rDNA, wingless, abdominal-A, and long-wavelength rhodopsin genes) and 144 morphological characters, both separately and in combination. Data were gathered from a representative set of forty-nine pseudomyrmecine species, plus eighteen species from various outgroups. There was substantial agreement among the results obtained from different datasets, and from different methods of phylogenetic inference (parsimony, Bayesian inference). The monophyly of the following groups is strongly supported (100% bootstrap support and 1.00 posterior probability in the molecular dataset): Pseudomyrmecinae, Pseudomyrmex, and Pseudomyrmex + Myrcidris. The status of the genus Tetraponera is less clear: the DNA sequence data indicate that the genus is paraphyletic, but morphological features and a unique insertion in the 28S gene support the monophyly of this taxon. Seven of nine Pseudomyrmex species groups, established previously on the basis of morphology alone, are strongly upheld, but monophyly is rejected for the P. pallens group and the P. viduus group. In the latter case, molecular evidence indicates the existence of two independent clades, associated with the ant-plants Triplaris and Tachigali, respectively, whose convergent morphological features had caused them to be placed erroneously in the same species group. The present results confirm an earlier assertion that obligate associations with domatia-bearing plants have arisen at least twelve times in the subfamily. Molecular and morphological data support the hypothesis of a sister-group relationship between Pseudomyrmecinae and Myrmeciinae (84% parsimony bootstrap, combined dataset), which implies a Cretaceous origin of the stem-group pseudomyrmecines in the southern hemisphere. Pseudomyrmecines appear to have arisen in the Palaeotropics and later dispersed from Africa to South America, where they experienced a pronounced burst of diversification.     

Phylogenetic relationships of flesh flies in the subfamily Sarcophaginae based on three mtDNA fragments (Diptera: Sarcophagidae)

In an effort to improve our knowledge of the phylogenetic relationships among species and genera of the subfamily Sarcophaginae, we analysed data from three mitochondrial gene fragments. Sequence data for portions of the genes cytochrome oxidase I (COI), cytochrome oxidase II (COII) and dehydrogenase subunit 4 (ND4) were obtained from 43 species of Sarcophagidae representing 15 genera. We used a Bayesian approach to simultaneously choose how best to partition the data and which substitution model to apply to each partition. Phylogenetic relationships were inferred using Bayesian Inference and Maximum Likelihood methods. Our results are consistent with monophyly of the subfamily Sarcophaginae (posterior probability 1; bootstrap support 93%), as well as with monophyly of several genera within the Sarcophaginae (including Sarcophaga s.l.; posterior probability 1; bootstrap support 97%). We found support for a sister‐group relationship between Ravinia Robineau‐Desvoidy and Oxysarcodexia Townsend, which has been hypothesised by past authors on the basis of morphological similarities, although this was supported only in the Bayesian analyses (posterior probability 0. 81–0. 98), and for some novel supra‐generic clades. Contrary to a recent morphological hypothesis, we do not find Helicobia Coquillett to be nested within Sarcophaga Meigen; our data suggest, but do not strongly support, a hypothesis that Peckia Robineau‐Desvoidy is the sister group to Sarcophaga.     

Morphological evolution and ecological diversification of the forest-dwelling poppies (Papaveraceae: Chelidonioideae) as deduced from a molecular phylogeny of the ITS region

Sequences of the ITS region of nrDNA were analyzed for the seven genera of Papaveraceae subf. Chelidonioideae s.str. Three major clades can be recognized. These are 1.Chelidonium/Hylomecon/Stylophorum, 2.Eomecon/Sanguinaria, and 3.Bocconia/Macleaya. The monophyly of genera in the first of these three clades is doubtful, and clades two and three are sister to each other. Use of the ITS phylogeny of the subfamily to trace its morphological and ecological evolution shows that morphological change is concentrated in theBocconia/Macleaya clade, and probably related to the evolution of wind-pollination from insect-pollination in these two genera after habitat shift.     

Examining monophyly in a large radiation of Madagascan butterflies (Lepidoptera: Satyrinae: Mycalesina) based on mitochondrial DNA data

The satyrine butterfly subtribe Mycalesina has undergone one of the more spectacular evolutionary radiations of butterflies in the Old World tropics. Perhaps the most phenotypically pronounced diversification of the group has occurred in the Malagasy region, where 68 currently recognized species are divided among five genera. Here, we report the results of phylogenetic analyses of sequence data from the cytochrome c oxidase II and cytochrome b mitochondrial genes, for a total of 54 mycalesine taxa, mostly from Madagascar. These molecular data complement an existing data set based on male morphological characters. The molecular results support the suggestion from morphology that three of the five Malagasy genera are paraphyletic and support the monophyly of at least three major morphological clades. Novel hypotheses of terminal taxon pairs are generated by the molecular data. Dense taxon sampling appears to be crucial for elucidating phylogenetic relationships within this large radiation. A potentially complex scenario for the origin of Malagasy mycalesines is proposed.     

Phylogeny of the North American vaejovid scorpion subfamily Syntropinae Kraepelin, 1905, based on morphology,mitochondrial and nuclear DNA

The first rigorous analysis of the phylogeny of the North American vaejovid scorpion subfamily Syntropinae is presented. The analysis is based on 250 morphological characters and 4221 aligned DNA nucleotides from three mitochondrial and two nuclear gene markers, for 145 terminal taxa, representing 47 species in 11 ingroup genera, and 15 species in eight outgroup genera. The monophyly and composition of Syntropinae and its component genera, as proposed by Soleglad and Fet, are tested. The following taxa are demonstrated to be para‐ or polyphyletic: Smeringurinae; Syntropinae; Vaejovinae; Stahnkeini; Syntropini; Syntropina; Thorelliina; Hoffmannius; Kochius; and Thorellius. The spinose (hooked or toothed) margin of the distal barb of the sclerotized hemi‐mating plug is demonstrated to be a unique, unambiguous synapomorphy for Syntropinae, uniting taxa previously assigned to different subfamilies. Results of the analysis demonstrate a novel phylogenetic relationship for the subfamily, comprising six major clades and 11 genera, justify the establishment of six new genera, and they offer new insights about the systematics and historical biogeography of the subfamily, and the information content of morphological character systems.