mtDNA Gene Sequences Research Articles

Phylogenetic Pattern, Evolutionary Processes and Species Delimitation in the Genus Echinococcus.

An accurate and stable alpha taxonomy requires a clear conception of what constitutes a species and agreed criteria for delimiting different species. An evolutionary or general lineage concept defines a species as a single lineage of organisms with a common evolutionary trajectory, distinguishable from other such lineages. Delimiting evolutionary species is a two-step process. In the first step, phylogenetic reconstruction identifies putative species as groups of organisms that are monophyletic (share a common ancestor) and exclusive (more closely related to each other than to organisms outside the group). The second step is to assess whether members of the group possess genetic exchangeability (where cohesion is maintained by gene flow among populations) or ecological exchangeability (where cohesion is maintained because populations occupy the same ecological niche). Recent taxonomic reviews have recognized nine species within the genus Echinococcus. Phylogenetic reconstructions of the relationships between these putative species using mtDNA and nuclear gene sequences show that for the most part these nine species are monophyletic, although there are important incongruences that need to be resolved. Applying the criteria of genetic and ecological exchangeability suggests that seven of the currently recognized species represent evolutionarily distinct lineages. The species status of Echinococcus canadensis and Echinococcus ortleppi could not be confirmed. Coalescent-based analyses represent a promising approach to species delimitation in these closely related taxa. It seems likely, from a comparison of sister species groups, that speciation in the genus has been driven by geographic isolation, but biogeographic scenarios are largely speculative and require further testing.

Genetic variation between and within Triodontophorus brevicauda and Triodontophorus nipponicus revealed by analyses of mtDNA and rDNA gene sequences

Triodontophorus spp. parasitizes the large intestine of equine, causing strongylid diseases. The present study assessed genetic variation in five gene regions within and between Triodontophorus brevicauda and Triodontophorus nipponicus from Heilongjiang Province and the Inner Mongolia Autonomous region. The five gene markers were three mitochondrial (mt) genes, cytochrome c oxidase subunit I (cox1), NADH dehydrogenase subunit 5 (nad5), cytochrome b (cytb); and two ribosomal RNA genes, the internal transcribed spacer 1 (ITS1) and the internal transcribed spacer 2 (ITS2). Partial (p) sequences of cox1, nad5, cytb and the complete ITS rDNA region were PCR amplified from individual nematodes, and the amplicons were subjected to sequencing in both directions. The size of the three mt genes is identical in both species: 761 bp (p cox1), 505 bp (pnad5) and 562 bp (pcytb); the length of the two ribosomal genes is different: 376 bp and 370 bp (ITS1), and 333 bp and 322 bp (ITS2), respectively. Intraspecific variation between T. brevicauda and T. nipponicus was 0–1.5% and 0–1.1% for pcox1, 0–2.0% and 0–2.0% for pnad5, 0–1.4% and 0–2.2% for pcytb, 0–0.8% and 0–1.1% for ITS1 and 0–0.9% and 0–2.2% for ITS2. Interspecific variation within the nematodes was 13.5–14.3% for pcox1, 15.5–18.7% for pnad5, 16.7–18.6% for pcytb, 11.5–13.1% for ITS1 and 16.0–18.4% for ITS2. Phylogenetic analyses based on the combined mt gene sequences, as well as with the ITS sequences, show each species forming a monophyletic group of individuals. However, samples of different species from the same geographical origin did not always cluster together. These results provide valuable information for further studies of systematics and population genetics of the genus Triodontophorus.

Sequencing of complete mitochondrial genome for Tsinling Tree Toad (Hyla tsinlingensis)

The complete mitochondrial genome sequence of Hyla tsinlingensis was determined in this research. The length of mitogenome is 17850 bp, including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, 1 OL and 1 control region. The phylogentic tree was reconstructed using the BI method based on concatenated nucleotide sequences of mtDNA genes (12S ribosomal small subunit gene/12S rRNA; NADH dehydrogenase subunit 1 gene/ND1, including adjacent transfer RNAs and the partial 16S ribosomal large subunit gene). The phylogenetic tree was split into two clades, Clade A and Clade B. The H. tsinlingensis which we determined clustered into Clade A.

Parasitism effects on white clover by root-knot and cyst nematodes and molecular separation of Heterodera daverti from H. trifolii

This research carried out an accurate identification of the root-knot and cyst-forming nematode species parasitizing white clover at the Laceno Lake area in Southern Italy. Two species, the root-knot nematode Meloidogyne hapla and the cyst nematode Heterodera daverti were identified by integrative taxonomic approaches (classical, isozyme pattern, and D2-D3 expansion segments of 28S rRNA, ITS rRNA and coxI of mtDNA gene sequences) and found parasitizing white clover roots. These nematodes were detected in stunted plants with a reduced number of rhizobium nodules and the host suitability was confirmed by the high nematode population densities ranging from 53 to 2350 eggs and J2s per g of fresh roots for M. hapla, and 1.36 eggs and J2s/cm3 of soil for H. daverti; and cyst nematode females were also detected on the roots of clover. The studies on the host-parasite relationships of nematode-feeding sites in white clover roots infected by these nematodes showed a high susceptible response. Meloidogyne hapla and H. daverti infections were also observed on nitrogen-fixing root nodules of white clover, where well established feeding sites allowed active nematode reproduction. Histological examination of nitrogen-fixing root nodule tissues revealed that the nematodes established their permanent feeding sites in the vascular bundles of nodules which appeared enlarged deformed and disorganised by the expansion of nematode feeding cells (giant cells and syncytium) and hyperplasia of the nodule cortex. Additionally, coxI of mtDNA gene is an efficient barcoding sequence for discriminating the identification of H. daverti from H. trifolii.

Probing the phylogenetic relationships of a few newly recorded intertidal zoanthids of Gujarat coast (India) with mtDNA COI sequences

The present study reports the phylogenetic relationship of six zoanthid species belonging to three genera, Isaurus, Palythoa, and Zoanthus identified using systematic computational analysis of mtDNA gene sequences. All six species are first recorded from the coasts of Kathiawar Peninsula, India. Genus: Isaurus is represented by Isaurus tuberculatus, genus Zoanthus is represented by Zoanthus kuroshio and Zoanthus sansibaricus, while genus Palythoa is represented by Palythoa tuberculosa, P. sp. JVK-2006 and Palythoa heliodiscus. Results of the present study revealed that among the various species observed along the coastline, a minimum of 99% sequence divergence and a maximum of 96% sequence divergence were seen. An interspecific divergence of 1–4% and negligible intraspecific divergence was observed. These results not only highlighted the efficiency of the COI gene region in species identification but also demonstrated the genetic variability of zoanthids along the Saurashtra coastline of the west coast of India.

Cryptic speciation in the white-shouldered antshrike (Thamnophilus aethiops, Aves – Thamnophilidae): The tale of a transcontinental radiation across rivers in lowland Amazonia and the northeastern Atlantic Forest

The growing knowledge on paleogeography and the recent applications of molecular biology and phylogeography to the study of the Amazonian biota have provided a framework for testing competing hypotheses of biotic diversification in this region. Here, we reconstruct the spatio-temporal context of diversification of a widespread understory polytypic Amazonian bird species (Thamnophilus aethiops) and contrast it with different hypotheses of diversification and the taxonomy currently practiced in the group. Sequences of mtDNA (cytochrome b and ND2) and nuclear (β-fibrinogen introns 5 and 7 and the Z-liked Musk4) genes, adding up to 4093bp of 89 individuals covering the Amazonian, Andean, and Atlantic Forest populations of T. aethiops were analyzed. Phylogenetic and population genetics analyses revealed ten reciprocally monophyletic and genetically isolated or nearly-isolated lineages in T. aethiops, highlighting several inconsistencies between taxonomy and evolutionary history in this group. Our data suggest that the diversification of T. aethiops started in the Andean highlands, and then proceeded into the Amazonian lowlands probably after the consolidation of the modern Amazonian drainage. The main cladogenetic events in T. aethiops may be related to the formation and structuring of large Amazonian rivers during the Late Miocene–Early Pleistocene, coinciding with the dates proposed for other lineages of Amazonian organisms. Population genetics data do not support climatic fluctuations as a major source of diversification in T. aethiops. Even though not entirely concordant with paleobiogeographic models derived from phylogenies of other vertebrate lineages, our results support a prominent role for rivers as major drivers of diversification in Amazonia, while underscoring that different diversification scenarios are probably related to the distinct evolutionary origins of groups being compared.

Phylogenetic relationships and timing of diversification in gonorynchiform fishes inferred using nuclear gene DNA sequences (Teleostei: Ostariophysi)

The Gonorynchiformes are the sister lineage of the species-rich Otophysi and provide important insights into the diversification of ostariophysan fishes. Phylogenies of gonorynchiforms inferred using morphological characters and mtDNA gene sequences provide differing resolutions with regard to the sister lineage of all other gonorynchiforms (Chanos vs. Gonorynchus) and support for monophyly of the two miniaturized lineages Cromeria and Grasseichthys. In this study the phylogeny and divergence times of gonorynchiforms are investigated with DNA sequences sampled from nine nuclear genes and a published morphological character matrix. Bayesian phylogenetic analyses reveal substantial congruence among individual gene trees with inferences from eight genes placing Gonorynchus as the sister lineage to all other gonorynchiforms. Seven gene trees resolve Cromeria and Grasseichthys as a clade, supporting previous inferences using morphological characters. Phylogenies resulting from either concatenating the nuclear genes, performing a multispecies coalescent species tree analysis, or combining the morphological and nuclear gene DNA sequences resolve Gonorynchus as the living sister lineage of all other gonorynchiforms, strongly support the monophyly of Cromeria and Grasseichthys, and resolve a clade containing Parakneria, Cromeria, and Grasseichthys. The morphological dataset, which includes 13 gonorynchiform fossil taxa that range in age from Early Cretaceous to Eocene, was analyzed in combination with DNA sequences from the nine nuclear genes and a relaxed molecular clock to estimate times of evolutionary divergence. This “tip dating” strategy accommodates uncertainty in the phylogenetic resolution of fossil taxa that provide calibration information in the relaxed molecular clock analysis. The estimated age of the most recent common ancestor (MRCA) of living gonorynchiforms is slightly older than estimates from previous node dating efforts, but the molecular tip dating estimated ages of Kneriinae (Kneria, Parakneria, Cromeria, and Grasseichthys) and the two paedomorphic lineages, Cromeria and Grasseichthys, are considerably younger.

European newts establish in Australia, marking the arrival of a new amphibian order

We document the successful establishment of a European newt (Lissotriton vulgaris) in south-eastern Australia, the first recorded case of a caudate species establishing beyond its native geographic range in the southern hemisphere. Field surveys in south-eastern Australia detected L. vulgaris at six sites, including four sites where the species had been detected 15 months earlier. Larvae were detected at three sites. Individuals had identical NADH dehydrogenase subunit 2 and cytb mtDNA gene sequences, and comparisons with genetic data from the species’ native range suggest that these individuals belong to the nominal subspecies L. v. vulgaris. Climatic conditions across much of southern Australia are similar to those experienced within the species’ native range, suggesting scope for substantial range expansion. Lissotriton vulgaris had been available in the Australian pet trade for decades before it was declared a ‘controlled pest animal’ in 1997, and thus the invasion documented here likely originated via the release or escape of captive animals. Lissotriton vulgaris is the sole member of an entire taxonomic order to have established in Australia, and given the potential toxicity of this species, further work is needed to delimit its current range and identify potential biodiversity impacts.

Morphological and molecular characterisation of the Saratov population of the European dagger nematode, Xiphinema diversicaudatum (Nematoda: Dorylaimida), with notes on phylogeography of the species

Plesiotype specimens of the European dagger nematode, Xiphinema diversicaudatum, were collected in Saratov, Russia, and morphologically, morphometrically and molecularly characterised. PCR with specific primer was developed for rapid diagnostics of this species. Phylogenetic relationships of X. diversicaudatum with other Xiphinema species, as inferred from the analyses of the D2-D3 expansion segments of 28S rRNA, ITS rRNA and coxI mtDNA gene sequences, are also provided. The study revealed that the clade X. diversicaudatum, together with X. bakeri (North America) and X. chengi (Asia), is related to Xiphinema species from the Mediterranean region and likely originated from a common ancestor inhabiting this area. The phylogenetic reconstructions with coxI mtDNA were used to describe the pattern of present genetic diversity of X. diversicaudatum and infer its biogeographical history in Europe. Haplotype diversity of X. diversicaudatum populations from Central Europe was significantly higher than those from south-eastern Europe, central, south-eastern and southern European Russia. It has been hypothesised that, during the Last Glacial Maximum, X. diversicaudatum populations probably persisted in refuge areas in the Carpathian mountains or the Alps, the species subsequently expanding from these areas and colonising other regions in Europe.

A Brave New World for an Old World Pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil

The highly polyphagous Old World cotton bollworm Helicoverpa armigera is a quarantine agricultural pest for the American continents. Historically H. armigera is thought to have colonised the American continents around 1.5 to 2 million years ago, leading to the current H. zea populations on the American continents. The relatively recent species divergence history is evident in mating compatibility between H. zea and H. armigera under laboratory conditions. Despite periodic interceptions of H. armigera into North America, this pest species is not believed to have successfully established significant populations on either continent. In this study, we provide molecular evidence via mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) partial gene sequences for the successful recent incursion of H. armigera into the New World, with individuals being detected at two sites (Primavera do Leste, Pedra Preta) within the State of Mato Grosso in Brazil. The mtDNA COI and Cyt b haplotypes detected in the Brazilian H. armigera individuals are common throughout the Old World, thus precluding identification of the founder populations. Combining the two partial mtDNA gene sequences showed that at least two matrilines are present in Brazil, while the inclusion of three nuclear DNA Exon-Primed Intron-Crossing (EPIC) markers identified a further two possible matrilines in our samples. The economic, biosecurity, resistance management, ecological and evolutionary implications of this incursion are discussed in relation to the current agricultural practices in the Americas.

A comprehensive molecular phylogeny for the hornbills (Aves: Bucerotidae)

The hornbills comprise a group of morphologically and behaviorally distinct Palaeotropical bird species that feature prominently in studies of ecology and conservation biology. Although the monophyly of hornbills is well established, previous phylogenetic hypotheses were based solely on mtDNA and limited sampling of species diversity. We used parsimony, maximum likelihood and Bayesian methods to reconstruct relationships among all 61 extant hornbill species, based on nuclear and mtDNA gene sequences extracted largely from historical samples. The resulting phylogenetic trees closely match vocal variation across the family but conflict with current taxonomic treatments. In particular, they highlight a new arrangement for the six major clades of hornbills and reveal that three groups traditionally treated as genera (Tockus, Aceros, Penelopides) are non-monophyletic. In addition, two other genera (Anthracoceros, Ocyceros) were non-monophyletic in the mtDNA gene tree. Our findings resolve some longstanding problems in hornbill systematics, including the placement of ‘Penelopides exharatus’ (embedded in Aceros) and ‘Tockus hartlaubi’ (sister to Tropicranus albocristatus). We also confirm that an Asiatic lineage (Berenicornis) is sister to a trio of Afrotropical genera (Tropicranus [including ‘Tockus hartlaubi’], Ceratogymna, Bycanistes). We present a summary phylogeny as a robust basis for further studies of hornbill ecology, evolution and historical biogeography.

Uncovering historical signature of mitochondrial DNA hidden in the nuclear genome: the biogeography of Schistocerca revisited.

Inadvertent coamplification of nuclear mitochondrial pseudogenes (numts) is a serious problem in mitochondrial systematics, but numts can also be a valuable source of information because they represent ancient forms of mtDNA. We present a conceptual framework of numt accumulation, which states that in a given species there can be two types of numts, synaponumts and autaponumts, resulting from integration occurring respectively before and after a speciation event. In a given clade, a species that diverged early can only have its own autaponumts as well as synaponumts that were already present in the genome of the last common ancestor. A species that diverged more recently may, however, have many different synaponumts integrated at each different divergence as well as its own autaponumts. Therefore it is possible to decipher the evolutionary history of a species based on the phylogenetic distribution of numts in a simultaneous analysis of numts and extant mtDNA. In this study, we test this idea empirically in the context of addressing a controversial question regarding the biogeography of the grasshopper genus Schistocerca Stål (Orthoptera: Acrididae), based on numts of the cytochrome c oxidase subunit I (COI) gene. We find that our empirical data can be explained adequately by our conceptual framework, and that the phylogenetic distribution of COI numts reveals intricate evolutionary histories about past speciation events that are otherwise difficult to detect using conventional markers. Our study strongly favours the Old World origin of the desert locust, Schistocerca gregaria and the New World Schistocerca species are descendants from an ancestral gregaria-like species that colonized the New World via westward transatlantic flight. However, the phylogenetic distribution of S. gregaria numts raises a distinct possibility that there might have been multiple founding events from Africa to America to give rise to the present-day diversity of the genus. This is a case study for a creative use of numts as molecular fossils, and we demonstrate that numts provide an interesting and powerful phylogenetic signal, much more than what extant mtDNA or nuclear gene sequences might be able to provide.

Population genetic structure of taimen, Hucho taimen (Pall.), in China

Hucho taimen (Pall.) is one of the largest salmonid species, and has been classified as an endangered species in China since 1998. In recent years, its wild stocks, habitats, and spawning grounds have been severely damaged by environmental degradation and human intervention. To protect and exploit this treasured species effectively and sustainably, the genetic diversity and structure of taimen populations in China was investigated using AFLP, microsatellite markers, and partial sequences of mtDNA genes. This paper summarizes these studies on the genetic diversity and structure of taimen populations in China.

Morphological and molecular characterisation and diagnostics of some species of Scutellonema Andrássy, 1958 (Tylenchida: Hoplolaimidae) with a molecular phylogeny of the genus

Scutellonema spp. are widely distributed across tropical and subtropical regions of the world and are associated with numerous agricultural and horticultural crops. Identification of many Scutellonema species is not always reliable, in part because many species share very similar diagnostic characters. In this study, we provide morphological and molecular characterisation of S. brachyurus from the USA and South Africa, S. bradys from Nigeria and three unidentified species from California, USA, New Zealand and Burkina Faso. Morphological descriptions, measurements, light and scanning electron microscopic photos and drawings are given for S. brachyurus. Females of S. brachyurus from the USA (type A) and South Africa (type B) showed a significant variation in the number of sectors and blocks on the lip annuli, ranging from about 4-12 and from 8-20, respectively. Molecular analysis using the D2-D3 of 28S rRNA, ITS rRNA and the COI mtDNA gene sequences revealed two distinct genotypes within S. brachyurus samples: type A (samples from USA, Italy, Korea, Taiwan) and type B (South Africa). Multivariate analyses determined that S. brachyurus from the USA and Taiwan (type A) differed from that from South Africa (type B) mainly in body, tail and DGO lengths, and ratios b′, c′, c and V. Phylogenetic relationships within Scutellonema are given as inferred from the analyses of the D2-D3 of 28S rRNA, ITS rRNA and the COI mtDNA gene sequences. PCR-RFLP diagnostic profiles and PCR with species-specific primers are developed for the studied Scutellonema species.

Free from mitochondrial DNA: Nuclear genes and the inference of species trees among closely related darter lineages (Teleostei: Percidae: Etheostomatinae)

Investigations into the phylogenetics of closely related animal species are dominated by the use of mitochondrial DNA (mtDNA) sequence data. However, the near-ubiquitous use of mtDNA to infer phylogeny among closely related animal lineages is tempered by an increasing number of studies that document high rates of transfer of mtDNA genomes among closely related species through hybridization, leading to substantial discordance between phylogenies inferred from mtDNA and nuclear gene sequences. In addition, the recent development of methods that simultaneously infer a species phylogeny and estimate divergence times, while accounting for incongruence among individual gene trees, has ushered in a new era in the investigation of phylogeny among closely related species. In this study we assess if DNA sequence data sampled from a modest number of nuclear genes can resolve relationships of a species-rich clade of North American freshwater teleost fishes, the darters. We articulate and expand on a recently introduced method to infer a time-calibrated multi-species coalescent phylogeny using the computer program *BEAST. Our analyses result in well-resolved and strongly supported time-calibrated darter species tree. Contrary to the expectation that mtDNA will provide greater phylogenetic resolution than nuclear gene data; the darter species tree inferred exclusively from nuclear genes exhibits a higher frequency of strongly supported nodes than the mtDNA time-calibrated gene tree.

Biobanked Amphibian Samples Confirmed To Species Level Using 16S rRNA DNA Barcodes.

The DNA barcoding technique is often used as a tool for validating species identity in biobanks. In the case of amphibians, the mitochondrial DNA (mtDNA) 16S ribosomal RNA (rRNA) gene is reported to fulfill the requirements of a universal DNA barcoding marker. The 16S primers are designed to specifically bind to the 16S rRNA gene, which is a very well-conserved mtDNA gene sequence in amphibians. DNA was extracted from thirteen known but different species of amphibians within the Zoological Society of London/Amphibian Ark's cryobank. After this, the DNA was amplified and analyzed by (1) the traditional DNA barcoding procedure that involves conventional polymerase chain reaction (PCR) and DNA sequencing and (2) a novel procedure, involving real-time PCR and melting temperatures. Both procedures used the same 16S primers. Successful DNA amplification and validation to the species or genus level was achieved in 10 out the 13 cases using the traditional approach. Nevertheless, after real-time PCR and melting temperature analysis, some variability was found between Common Frog samples but more concerning, the same melting temperature was recorded in unrelated species (Common Toad, Common Frog and Amazon Milk Frog), despite their 16S sequences exhibiting a high degree of variability. We conclude that traditional DNA barcoding using 16S rRNA sequences is suitable for validating the specific identity of amphibian samples within biobanks and that modification of the current 16S real-time PCR and melting temperature analysis is required before it can be employed as a cheaper and faster alternative.

Taxonomic Relationships among Turkish Water Frogs as Revealed by Phylogenetic Analyses using mtDNA Gene Sequences

We assessed taxonomic relationships among Turkish water frogs through estimation of phylogenetic relationships among 62 adult specimens from 44 distinct populations inhabiting seven main geographical regions of Turkey using 2897 bp sequences of the mitochondrial Cytb, 12S rRNA and 16S rRNA genes with equally-weighted parsimony, likelihood, and Bayesian methods of inference. Monophyletic clade (Clade A) of the northwesternmost (Thrace) samples is identified as Pelophylax ridibundus. The other clade (Clade B) consisted of two monophyletic subclades. One of these contains specimens from southernmost populations that are regarded as an unnamed species. The other subclade consists of two lineages, of which one corresponds to P. caralitanus and another to P. bedriagae. Taxonomic relationships of these two species are discussed and recognition of P. caralitanus as a subspecies of P. bedriagae is proposed.

Monophyly of clade III nematodes is not supported by phylogenetic analysis of complete mitochondrial genome sequences

BackgroundThe orders Ascaridida, Oxyurida, and Spirurida represent major components of zooparasitic nematode diversity, including many species of veterinary and medical importance. Phylum-wide nematode phylogenetic hypotheses have mainly been based on nuclear rDNA sequences, but more recently complete mitochondrial (mtDNA) gene sequences have provided another source of molecular information to evaluate relationships. Although there is much agreement between nuclear rDNA and mtDNA phylogenies, relationships among certain major clades are different. In this study we report that mtDNA sequences do not support the monophyly of Ascaridida, Oxyurida and Spirurida (clade III) in contrast to results for nuclear rDNA. Results from mtDNA genomes show promise as an additional independently evolving genome for developing phylogenetic hypotheses for nematodes, although substantially increased taxon sampling is needed for enhanced comparative value with nuclear rDNA. Ultimately, topological incongruence (and congruence) between nuclear rDNA and mtDNA phylogenetic hypotheses will need to be tested relative to additional independent loci that provide appropriate levels of resolution.ResultsFor this comparative phylogenetic study, we determined the complete mitochondrial genome sequences of three nematode species, Cucullanus robustus (13,972 bp) representing Ascaridida, Wellcomia siamensis (14,128 bp) representing Oxyurida, and Heliconema longissimum (13,610 bp) representing Spirurida. These new sequences were used along with 33 published nematode mitochondrial genomes to investigate phylogenetic relationships among chromadorean orders. Phylogenetic analyses of both nucleotide and amino acid sequence datasets support the hypothesis that Ascaridida is nested within Rhabditida. The position of Oxyurida within Chromadorea varies among analyses; in most analyses this order is sister to the Ascaridida plus Rhabditida clade, with representative Spirurida forming a distinct clade, however, in one case Oxyurida is sister to Spirurida. Ascaridida, Oxyurida, and Spirurida (the sampled clade III taxa) do not form a monophyletic group based on complete mitochondrial DNA sequences. Tree topology tests revealed that constraining clade III taxa to be monophyletic, given the mtDNA datasets analyzed, was a significantly worse result.ConclusionThe phylogenetic hypotheses from comparative analysis of the complete mitochondrial genome data (analysis of nucleotide and amino acid datasets, and nucleotide data excluding 3rd positions) indicates that nematodes representing Ascaridida, Oxyurida and Spirurida do not share an exclusive most recent common ancestor, in contrast to published results based on nuclear ribosomal DNA. Overall, mtDNA genome data provides reliable support for nematode relationships that often corroborates findings based on nuclear rDNA. It is anticipated that additional taxonomic sampling will provide a wealth of information on mitochondrial genome evolution and sequence data for developing phylogenetic hypotheses for the phylum Nematoda.

Complete sequence and gene organization of the mitochondrial genome of scaly-sided merganser (Mergus squamatus) and phylogeny of some Anatidae species

The scaly-sided merganser (Mergus squamatus) is an endangered bird species on the IUCN Red List with the estimated global population of less than 2,500 individuals at present. In the present study, we studied the complete mitochondrial genome (mtDNA) and the phylogenetic of M. squamatus by PCR amplification and GenBank data. The genome was 16,595 bp in length and contained 37 genes (13 protein coding genes, two rRNAs, and 22 tRNAs) and a non-coding control region (D-loop). All protein-coding genes of M. squamatus mtDNA start with a typical ATG codon, except ND1, COI, and COII uses GTG as their initial codon. TAA, T- and TAG as the terminate codon occurred very commonly in the sequence. All tRNA genes can be folded into canonical cloverleaf secondary structure except for tRNA(Ser) (AGY) and tRNA(Leu) (CUN), which lose ''DHU'' arm. The genome sequences had been deposited in GenBank under accession number HQ833701. Based on the concatenated nucleotide sequences of mtDNA genes (Cyt b and D-loop), we reconstructed phylogenetic trees and discussed the phylogenetic relationships among ten Anatidae species. The results are different from the present classification, and we support Lophodytes cucullatus and Mergullus albellus to be members of the genus Mergus.

Cytotaxonomy and DNA taxonomy of lizards (Squamata, Sauria) from a tropical dry forest in the Chamela-Cuixmala Biosphere Reserve on the coast of Jalisco, Mexico

Tropical dry forests contribute to a substantial proportion of the herpetological diversity of Mexico. The south-western coast of Jalisco is one of the more important areas by number of endemics and the high presence of endangered and restricted species. In this paper we used a combined karyological and molecular genetic (sequences of mtDNA genes for NDH2, cytb or 16S rDNA) approach to genetically characterize 13 lizard species belonging to seven families that inhabit the dry forests of the Chamela-Cuixmala Biosphere Reserve (Anguidae: Gerrhonotus cf. liocephalus; Eublepharidae: Coleonyx elegans; Phyllodactylidae: Phyllodactylus lanei; Gekkonidae: Hemidactylus frenatus; Phrynosomatidae: Sceloporus melanorhinus, S. utiformis, Urosaurus bicarinatus; Polychrotidae: Norops nebulosus; Scincidae: Mabuya unimarginata, Plestiodon parvulus; Teiidae: Ameiva undulata, Aspidoscelis communis, A. lineattissima). The karyotypes of six species were here described for the first time (G. liocephalus, 2n = 38, 14 macrochromosomes and 24 microcromosomes; C. elegans, 2n = 24 FN = 26; N. nebulosus 2n = 30, 13 macroand 17 microchromosomes; M. unimarginata 2n = 32, 18 macroand 14 microchromosomes; P. parvulus 2n = 26, 12 macroand 14 microchromosomes; A. undulata 2n = 50, 26 macroand 24 microchromosomes). Chromosomal heteromorphism was found in C. elegans, N. nebulosus, and S. melanorhinus. For P. lanei we found a karyotype different from that previously described in other localities. This variation matched with a high genetic divergence usually found in different species. The DNA typing of mtDNA genes allowed the identification of the taxonomic affinities of five Mexican endemic species, namely: U. bicarinatus, A. nebulosus, P. parvulus, A. lineattissima and A. communis. The specimen of Gerrhonotus from Chamela is very divergent by 16S rDNA and probably does not belong to the so far studied species of Gerrhonotus. High genetic divergence has been also observed between samples of A. undulata and U. bicarinatus from different regions. In these latter two cases, additional data are needed to understand the taxonomic status of these populations.