A phylogeny of the damselfly genus calopteryx (Odonata) using mitochondrial 16S rDNA markers

We seek to reconstruct the phylogenetic relationships of the damselfly genus Calopteryx, for which extensive behavioral and morphological knowledge already exists. To date, analyses of the evolutionary pathways of different life history traits have been hampered by the absence of a robust phylogeny based on morphological data. In this study, we concentrate on establishing phylogenetic information from parts of the 16S rDNA gene, which we sequenced for nine Calopteryx species and five outgroup species. The mt 16S rDNA data set did not show signs of saturated variation for ingroup taxa, and phylogenetic reconstructions were insensitive to variation of outgroup taxa. Parsimony, neighbor-joining, and maximum-likelihood reconstructions agreed on parts of the tree. A consensus tree summarizes the significant results and indicates problematic nodes. The 16S rDNA sequences support monophyly of the genera Mnais, Matrona, and Calopteryx. However, the genus Calopteryx may not be monophyletic, since Matrona basilaris and Calopteryx atrata are sister taxa under every parameter setting. The North American and European taxa each appear as monophyletic clades, while the Asian Calopteryx atrata and Calopteryx cornelia are not monophyletic. Our data implies a different paleobiogeographic history of the Eurasian and North American species, with extant Eurasian species complexes shaped by glacial periods, in contrast to extant North American species groups.     

A molecular phylogeny of the genus Ichthyocotylurus (Digenea, Strigeidae)

Three nucleotide data sets, two nuclear (ribosomal internal transcribed spacer regions 1 and 2, ITS1 and ITS2) and one mitochondrial (cytochrome c oxidase subunit 1, CO1), were analysed using distance matrix and maximum likelihood methods to determine the inter-relationships amongst the four species attributed to the genus Ichthyocotylurus Odening, 1969. Sequence data obtained from all gene loci investigated supported the position of Ichthyocotylurus variegatus as a species discrete from Ichthyocotylurus platycephalus. Phylogenetic analyses yielded congruent trees, with I. variegatus isolates comprising a common clade to which I. platycephalus constitutes a sister taxon. Ichthyocotylurus erraticus and Ichthyocotylurus pileatus were found to demonstrate a similarly close inter-specific relationship. The greatest intra-generic divergence occurred in the CO1 region (16% variability), with resultant disparities in three to eight encoded amino acids. PCR amplification yielded multiple ITS1 products for all Ichthyocotylurus spp. Analyses of equivalent-sized amplicons showed 5.4% intra-generic variation and several point mutations between I. variegatus isolates from different geographical localities and from different piscine hosts. The ITS2 locus was extremely conserved, with less than 1% variation between species. No intra-specific variation was recorded for any CO1 or ITS2 sequences.     

Cryptic diversity in an endemic rainforest skink (Gnypetoscincus queenslandiae)

Gynpetoscincus is a monotypic genus of skink endemic to the ancient and biogeographically complex wet tropical rainforests of northern Queensland. Surveys of variation in mitochondrial DNA and allozymes inG. queenslandiae revealed a major genetic break within the skink's 275 km range, clearly dividing it into northern and southern populations. The genetic differences between the two groups of populations are several-fold greater than differences between similarly separated localities within the southern group. The presence of two strongly differentiated evolutionary lineages within this narrowly distributed species highlights the value of genetic surveys to estimate evolutionary diversity in old, biogeographically complex biomes such as these rainforests.     

A phylogeny of Chinese species in the genus Phrynocephalus (Agamidae) inferred from mitochondrial DNA sequences

We investigated the phylogenetic relationships among most Chinese species of lizards in the genus Phrynocephalus (118 individuals collected from 56 populations of 14 well-defined species and several unidentified specimens) using four mitochondrial gene fragments (12S rRNA, 16S rRNA, cytochrome b, and ND4-tRNA(LEU)). The partition-homogeneity tests indicated that the combined dataset was homogeneous, and maximum-parsimony (MP), neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian (BI) analyses were performed on this combined dataset (49 haplotypes including outgroups for 2058bp in total). The maximum-parsimony analysis resulted in 24 equally parsimonious trees, and their strict consensus tree shows that there are two major clades representing the Chinese Phrynocephalus species: the viviparous group (Clade A) and the oviparous group (Clade B). The trees derived from Bayesian, ML, and NJ analyses were topologically identical to the MP analysis except for the position of P. mystaceus. All analyses left the nodes for the oviparous group, the most basal clade within the oviparous group, and P. mystaceus unresolved. The phylogenies further suggest that the monophyly of the viviparous species may have resulted from vicariance, while recent dispersal may have been important in generating the pattern of variation among the oviparous species.     

A molecular phylogeny of the genus Gammarus (Crustacea: Amphipoda) based on mitochondrial and nuclear gene sequences

A phylogeny of the genus Gammarus Fabricius, 1775 was constructed using DNA sequence data from the mitochondrial genes COI and 16S, and the nuclear genes 18S and 28S. Both parsimony and Bayesian analyses were conducted on separate and combined data partitions. The Bayesian phylogeny from the combined analysis was selected as the preferred phylogenetic hypothesis. The hypothesis supports monophyly of the genus Gammarus, paraphyly of the European-North American Gammarus, and monophyly of the Asian Gammarus. The Asian clade was further split into a southeastern group and a northwestern group. The dramatic climate change following the uplift of the Tibetan Plateau was probably the most important factor in triggering the diversification of southeastern and northwestern groups. The genus Sinogammarus is invalid and should be part of the genus Gammarus.     

Molecular phylogeny of the speciose vole genus Microtus (Arvicolinae, Rodentia) inferred from mitochondrial DNA sequences

Voles of the genus Microtus represent one of the most speciose mammalian genera in the Holarctic. We established a molecular phylogeny for Microtus to resolve contentious issues of systematic relationships and evolutionary history in this genus. A total of 81 specimens representing ten Microtus species endemic to Europe as well as eight Eurasian, six Asian and one Holarctic species were sequenced for the entire cytochrome b gene (1140 bp). A further 25 sequences were retrieved from GenBank, providing data on an additional 23, mainly Nearctic, Microtus species. Phylogenetic analysis of these 48 species generated four well-supported monophyletic lineages. The genus Chionomys, snow voles, formed a distinct and well-supported lineage separate from the genus Microtus. The subgenus Microtus formed the strongest supported lineage with two sublineages displaying a close relationship between the arvalis species group (common voles) and the socialis species group (social voles). Monophyly of the Palearctic pitymyid voles, subgenus Terricola, was supported, and this subgenus was also subdivided into two monophyletic species groups. Together, these groupings clarify long-standing taxonomic uncertainties in Microtus. In addition, the "Asian" and the Nearctic lineages reported previously were identified although the latter group was not supported. However, relationships among the main Microtus branches were not resolved, suggesting a rapid and potentially simultaneous radiation of a widespread ancestor early in the history of the genus. This and subsequent radiations discernible in the cytochrome b phylogeny, show the considerable potential of Microtus for analysis of historical and ecological determinants of speciation in small mammals. It is evident that speciation is an ongoing process in the genus and that the molecular data provides a vital insight into current species limits as well as cladogenic events of the past.     

Molecular phylogeny of the Oriental butterfly genus Arhopala (Lycaenidae,Theclinae) inferred from mitochondrial and nuclear genes

Abstract. We present a phylogeny for a selection of species of the butterfly genus Arhopala Boisduval, 1832 based on molecular characters. We sequenced 1778 bases of the mitochondrial genes Cytochrome Oxidase 1 and 2 including tRNA^Leu, and a 393‐bp fragment of the nuclear wingless gene for a total of 42 specimens of 33 species, representing all major species groups. Analyses of mtDNA and wingless genes show congruent phylogenetic signal. The phylogeny presented here confirms the monophyly of the centaurus, eumolphus, camdeo and epimuta groups and the amphimuta subgroup. It confirms close relationships between species within the agelastus group, that together with the amphimuta subgroup, centaurus and camdeo groups form a monophyletic group. However, incongruencies with previous taxonomic studies also occur; the amphimuta and silhetensis groups are not monophyletic, as is the genus Arhopala itself. One enigmatic species, A. kinabala, was evaluated further for topology and the support for basal placement of this species is due mainly to the wingless gene. However, in the Parsimony analysis, and subsequent Maximum Likelihood evaluations, certain nodes could not be resolved due to insufficient support. The mtDNA shows extreme AT bias with compositional heterogeneity at 3rd codon positions, which may result in saturation. By contrast, the wingless gene does not show compositional bias, suggesting that poor support is not due solely to saturation. The evaluation of morphological characters used in previous studies on Arhopala systematics on the molecular tree indicates that the macular pattern and the absence of tails at the hind wings show extensive homoplasy. A significant phylogenetic signal (as indicated by T‐PTP tests) is present in several of these morphological characters, which are nevertheless of limited use in phylogenetic studies due to their labile nature.     

A plastid DNA phylogeny of the genus Acacia Miller (Acacieae, Leguminoseae)

Past classifications of the tribe Acacieae Rchb. are outlined and the confusion concerning the relationships of the three subgenera of Acacia Mill. are highlighted. A plastid DNA analysis of Acacieae shows that the genus Acacia is not monophyletic. Furthermore subgenera Acacia Vassal and Aculeiferum Vassal are sister taxa and neither appear closely related to subgenus Phyllodineae (DC.) Ser. Subgenera Acacia and Aculeiferum form a clade that is basal to a well-supported clade consisting of tribe Ingeae Benth. taxa, Faidherbia albida (Del.) A. Chev. and subgenus Phyllodineae. The series of relationships suggested by the cpDNA data contradicts previous investigations of the tribe. Possible explanations of this conflict are explored, and the taxonomic implications of the plastid DNA data set are considered.     

Molecular phylogeny of the genus Mus (Rodentia: Murinae) based on mitochondrial and nuclear data

The genus Mus encompasses at least 38 species divided into four subgenera: Mus , Pyromys , Nannomys and Coelomys . The subgenus Mus , which comprises the house mouse and related species, is by far the most extensively studied, although the subgenus Nannomys is the most speciose. Although the relationships within the subgenus Mus are rather well characterized, those between subgenera are still unclear. In the present study, phylogenetic analyses of the whole genus were performed using a larger species sample of Nannomys than in previous studies, and a nuclear gene (IRBP) in addition to mitochondrial data (cytochrome b and 12S rRNA). Members of the Acomyinae and Murinae were used as outgroups. Separate and combined analyses were performed with maximum parsimony, maximum likelihood and Bayesian methods, and divergence times were estimated. The results showed that the monophyly of the genus Mus and of each subgenus was strongly supported by the three genes and the combined analysis. The phylogenies derived from the three genes were on the whole congruent; however, several conflicting topologies were observed such as the relationships between the three Asian species of the subgenus Mus ( caroli , cervicolor and cookii ). Increasing the taxonomic sampling of Nannomys did not satisfactorily improve the resolution of relationships between the four subgenera. In addition, molecular calibrations indicate that the Mus and Nannomys radiation coincided with major environmental changes.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 417–427.     

Molecular phylogeny of the genus Buteo (Aves: Accipitridae) based on mitochondrial marker sequences

DNA sequences of the mitochondrial nd6 gene and the non-repetitive part of the pseudo-control region (PsiCR) were isolated from 101 individuals to analyze the phylogenetic relationships among all buzzards of the genus Buteo and other buteonine genera. Comparisons of the two marker sequences indicate that the PsiCR evolved two times faster than the nd6 gene. The PsiCR proved to be an efficient, neutral genetic marker sequence for phylogenetic analyses at the intrageneric level, especially suitable for analyses based on old tissues, where only short fragments can be obtained. The molecular data set implies a neotropical origin of the genus Buteo. Monophyly of the genus Buteo as currently defined is contradicted due to the positions of Asturina nitida, Geranoaetus melanoleucus, Buteo magnirostris, and Buteo leucorrhous. These findings suggest several taxonomic consequences. A. nitida and G. melanoleucus should be included into the genus Buteo. Moreover, B. leucorrhous should be transferred into the genus Percnohierax (which clusters with Parabuteo), and B. magnirostris into the genus Rupornis. According to this classification of the genus Buteo, the basal lineage of the genus is formed by a clade containing Buteo polyosoma, Buteo poecilochrous, and Buteo melanoleucus. The "woodland buteos" form a paraphyletic assemblage with B. magnirostris as a clearly separated lineage basal to the genus Buteo.     

A phylogeny for grasshoppers of the genus Chitaura (Orthoptera: Acrididae) from Sulawesi, Indonesia, based on mitochondrial DNA sequence data

The Indonesian island of Sulawesi occupies a central position within the biogeographically complex region known as Wallacea. Its fauna is characterised by a high rate of endemism and a patchwork distribution of taxa within the island. The grasshopper genus Chitaura is a good example having at least ten endemic species with predominantly parapatric distributions. It can be used as a model for determining the origins of Sulawesi taxa and the within-island evolution that has led to the present patterns of distribution. Here we present a phylogenetic hypothesis for 28 individuals within the genus, including individuals of one species from Java and two from the Moluccas, based on DNA sequences of the mitochondrial cytochrome oxidase 1 gene. Frequent sequence heteroplasmy was observed. The phylogenetic hypothesis is consistent with recent interpretations of the geological history of Sulawesi suggesting separate evolution on the island for 7–14 Myr, possibly since South Sulawesi was connected to Borneo. Within the island, the pattern of genetic divergence is dominated by a strong correlation with geographic distance, with exceptions indicating past or present barriers to dispersal. Colonisation of the Moluccas from North or Central Sulawesi is implied. Levels of genetic divergence are compared with distribution patterns of colour morphs and with possible effects of tectonic movements in the Cenozoic, or Pleistocene climatic, vegetational and sea-level changes.     

Morphology,phylogeny and taxonomy of the genus Sectonema (Nematoda,Aporcelaimidae)

This contribution provides new insights in the phylogeny and taxonomy of the nematode genus Sectonema with an integrative approach. A brief historical outline of the matter is presented. Then, the morphological pattern of the genus is revised and illustrated, the nature of its stomatal protruding structure, either a reduced odontostyle or a mural tooth, being its most relevant diagnostic feature. The existence of cilia‐ or seta‐like structures in the perioral area and/or at the anterior part of cheilostom of some species is evidenced by SEM observations for the first time. Available molecular data (D2–D3 LSU‐rRNA gene) are analysed, including two new sequences of S. septentrionale from Spain. The monophyly of the genus is confirmed, and two species groups with geographical projection are tentatively identified. A close relationship with Metaporcelaimus is also demonstrated as both taxa constitute a highly supported clade. A likely polyphyly of the family Aporcelaimidae is once more demonstrated. Finally, an updated taxonomy of the genus is proposed, including revised diagnosis, list of species, identification key and a compendium of their main morphometrics.     

A global plastid phylogeny of the fern genus Asplenium (Aspleniaceae)

The infrageneric relationships and taxonomy of the largest fern genus, Asplenium (Aspleniaceae), have remained poorly understood. Previous studies have focused mainly on specific species complexes involving a few or dozens of species only, or have achieved a large taxon sampling but only one plastid marker was used. In the present study, DNA sequences from six plastid markers (atpB, rbcL, rps4, rps4-trnS, trnL and trnL-F) of 1030 accessions (616 of them newly sequenced here) representing c. 420 species of Asplenium (60% of estimated species diversity), 16 species of Hymenasplenium, three Diplaziopsidaceae, and four Rhachidosoraceae were used to produce the largest genus-level phylogeny yet for ferns. Our major results include: (i) Asplenium as broadly circumscribed is monophyletic based on our inclusion of representatives of 32 of 38 named segregate genera; (ii) 11 major clades in Asplenium are identified, and their relationships are mostly well-resolved and strongly supported; (iii) numerous species, unsampled in previous studies, suggest new relationships and numerous cryptic species and species complexes in Asplenium; and (iv) the accrued molecular evidence provides an essential foundation for further investigations of complex patterns of geographical diversification, speciation and reticulate evolution in this family.     

A molecular phylogeny of the hawkmoth genus Hyles (Lepidoptera: Sphingidae, Macroglossinae)

The hawkmoth genus Hyles is one of 15 genera in the subtribe Choerocampina of the subfamily Macroglossinae. Due to a remarkable uniformity, morphological characters usually used to identify and classify Lepidoptera at the species level cannot be used in this genus. Instead, we used DNA sequences comprising about 2300 bp derived from the mitochondrial genes COX I, COX II, and tRNA-leucine to elucidate the phylogeny of Hyles. The results corroborate the monophyly of Hyles but conflict with previous internal classifications of the genus based on morphology. Hyles seems to have evolved in the Neotropics during the Oligocene/Eocene epochs and the molecular data (which evolved clock-like) confirm the hypothesis that it is a very young genus that radiated on a global scale rather quickly. We hypothesize its sister group to be one of the genera Deilephila, Theretra or Xylophanes. The Nearctic may have been colonized rapidly by Hyles once the land bridge formed during the Pliocene, since within this same Epoch, the invasion of the Palaearctic appears to have proceeded from the East, via the Bering route. The colonization of Australia appears to have occurred rather early in Hyles radiation, although the route is not clear. We propose that the radiation of the Hyles euphorbiae-complex s. str. (HEC) occurred as recently as the Pliocene/Pleistocene boundary and that its roots can still be reconstructed in Asia. Hyles dahlii is closely related to the HEC, but a sister group relationship to the HEC s. str. cannot be corroborated unequivocally. HEC population ranges appear to have tracked climate oscillations during the Pleistocene Ice Ages, resulting in hybridization around the Mediterranean Sea as they repeatedly intermingled. Comparison of the phylogeny with food plant affiliations leads us to hypothesize that Euphorbia monophagy evolved at least two times independently within Hyles.     

A phylogeny of the northern temperate leafy liverwort genus Scapania (Scapaniaceae, Jungermanniales)

Scapania is a northern temperate genus with a few disjunctions in the south. Despite receiving considerable attention, the supraspecific classification of this genus remains unsatisfactorily solved. We use three molecular markers (nrITS, cpDNA trnL-F region, atpB-rbcL spacer) and 175 accessions belonging to 50 species (plus eight outgroup taxa) to estimate the phylogeny and to test current classification systems. Our data support the classification of Scapania into six rather than three subgenera, rearrangements within numerous sections, and inclusion of Macrodiplophyllum microdontum. Scapania species with a plicate perianth form three early diverging lineages; the most speciose subgenus, Scapania s.str., represents a derived clade. Most morphological species concepts are supported by the molecular topologies but classification of sect. Curtae requires further study. Southern lineages are nested in northern hemispheric clades. Palearctic-Nearctic distribution ranges are supported for several species.     

A molecular phylogeny of the cosmopolitan hyperdiverse genus Hydraena Kugelann (Coleoptera,Hydraenidae)

With almost 900 described species, Hydraena Kugelann (Hydraenidae) is one of the largest genera among Coleoptera. The subgeneric classification of Hydraena has been controversial, with 11 subgeneric names having so far been attributed to it. Some of these, Haenydra Rey and Spanglerina Perkins, have been treated as valid genera, as subgenera or as species groups. The most recent complete treatment of the genus, based on a cladistic analysis of morphological characters, recognized two major lineages, and only these were classified as subgenera: Hydraenopsis (mainly Gondwanan distribution), and Hydraena s.str. (mainly Laurasian). Here, we reconstruct the phylogeny of Hydraena using 212 species plus several outgroups and approximately 4 kb of sequence data from two nuclear (SSU and LSU) and four mitochondrial genes (cox1, rrnL, trnL and nad1). Data were aligned with two different strategies of multiple alignment (implemented in mafft and prank ), and the phylogenies reconstructed using maximum likelihood and Bayesian methods. We estimated approximate ages of the main nodes using a relaxed molecular clock with Bayesian methods, and an a priori evolutionary rate of 0.01 substitutions/site/million years (Ma) plus a calibration point based on a biogeographical split. We found strong support for the monophyly of Hydraena and many of the clades recognized with morphological data. The following clades are considered as subgenera: Phothydraena Kuwert, Spanglerina Perkins, Holcohydraena Kuwert, Hydraenopsis Janssens and Hydraena s.str. The placement of three species groups, two Neotropical (H. multispina group, H. paeminosa group) and one South African/Madagascan (H. monikae group), is uncertain, and they are considered incertae sedis within Hydraena. The origin of the genus was estimated to be in the Lower Eocene, with many species complexes diversifying in the Pleistocene. Dispersal events seem to have played a key role in order to determine the current distribution of the species groups in the southern hemisphere (mainly in Hydraenopsis).     

Molecular phylogeny and phylogeography of the Greek populations of the genus Orthometopon (Isopoda, Oniscidea) based on mitochondrial DNA sequences

We infer phylogenetic relationships among isopod species of the genus Orthometopon distributed in the Greek area, comparing partial mitochondrial DNA sequences for cytochrome oxidase I (COI). All phylogenetic analyses produced topologically identical trees that revealed a well-resolved phylogeny. These trees support the monophyly of the genus Orthometopon , and suggest two clades that correspond to separate geographical regions (west and east of the mid-Aegean trench). However, the phylogenetic relationships among Greek populations of Orthometopon spp. are different from the presumed pattern on the basis of morphological evidence. The distinct geographical distribution of the major clades of the phylogenetic tree and its topology suggest a spatial and temporal sequence of phylogenetic separations, which coincide with some major palaeogeographical separations during the geological history of the Aegean Sea. The results stress the need for a reconsideration of the evolutionary history of Orthometopon species, which will help overcome difficulties encountered in classical taxonomy at the species level. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 152 , 707–715.     

A test of a mitochondrial gene-based phylogeny of woodpeckers (genus Picoides) using an independent nuclear gene, beta-fibrinogen intron 7

A conservative estimate of the species tree for the woodpecker genus Picoides based on two mitochondrial protein-coding genes is tested using sequences of an independently evolving nuclear intron, beta-fibrinogen intron 7. The mitochondrial gene-based topology and the intron-based topology are concordant, and a partition-homogeneity statistical test did not detect phylogenetic heterogeneity. The intron evolves more slowly than the mitochondrial sequences and tends not to resolve relationships among recently evolved species. However, the intron is superior over mitochondrial genes in resolving older bifurcations in the phylogeny. The two data sets were combined resulting in a robust estimate of the Picoides species tree in which most every node is statistically supported by bootstrap proportions. The Picoides species tree clearly shows that many morphological and behavioral characters used to lump species into this single genus have evolved by convergent evolution. Picoides is considered the largest genus of woodpeckers, but the molecular-based species tree suggests that Picoides is actually a conglomerate of several smaller groups.     

A new genus and species of termites (Isoptera, Termitidae, Nasutitermitinae) from the rainforest of northern Peru

A new genus and species of nasutitermitine termites are described and illustrated, based on soldier and worker characters. Sinqasapatermesgen. n., can be distinguished from all other nasutitermitine genera by its singular worker gut coiling and enteric valve characters: distal margin of the enteric valve not everted into the paunch but bending towards the ileum, that is, directed against the flow of food; enteric valve armature with one ring of six equal subtriangularly-shaped ridges, each ridge with short spines on the entire surface; enteric valve armature situatedonexternal face of cone, facing the internal ileum wall; enteric valve seating tri-lobed and separated from remaining portionof the paunch; paunch subdivided. Sinqasapatermes sachaesp. n., was collected on a tree in a very narrow flattened tunnel that was well concealed beneath lichens in a northern Peru rainforest (Arcadia, Loreto Province).     

Charrs of the genus Salvelinus (Salmonidae): hybridization,phylogeny and evolution

Hydrobiologia - Evolutionary history, systematics and taxonomy of charrs of the genus Salvelinus and especially of the representatives of the S. alpinus–S. malma species complex remain...