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The complete mitochondrial genome of Anisakis simplex (Ascaridida: Nematoda) and phylogenetic implications.

We determined the nucleotide sequence of the complete mitochondrial genome of the nematode species Anisakis simplex. The genome is circular, 13,916 bp in size and conforms to the general characteristics of nematode mitochondrial DNAs. The gene arrangement of A. simplex is the same as that of Ascaris suum and almost identical to those of rhabditid species with a minor exception concerning the relative position of the AT-rich and non-coding regions and radically different from those of spirurid species. Along with comparisons of gene arrangement, phylogenetic analyses (maximum parsimony, neighbour joining and maximum likelihood methods) based on concatenated amino acid sequences of 12 protein-coding genes from 13 nematode species provided strong support for the sister-group relationship between Ascaridida and Rhabditida. The Shimodaira-Hasegawa and Templeton's tests both rejected the alternative hypothesis of a closer relationship between Ascaridida and Spirurida. These results contradicted the traditional view of nematode classification and a recent molecular phylogenetic study of 18S rDNA data that assigned Ascaridida and Spirurida as being a sister-group. Mapping of gene arrangement across the phylogenetic tree lead to the assumption that the conserved gene arrangement found in Ascaridida-Rhabditida members might have been acquired after the most recent common ancestor of ascaridid/rhabditid members branched off from the basal stock of the rhabditid lineage.

Amino Acid Sequence↗

Molecular systematics of the gonorynchiform fishes (Teleostei) based on whole mitogenome sequences: implications for higher-level relationships within the Otocephala.

Although the order Gonorynchiformes includes only 31 species assigned to seven genera and four families, it exhibits a large variety of anatomical structures, making difficult the reconstruction of phylogenetic relationships among its representatives. Within the basal teleosts, the Gonorynchiformes belong to the Otocephala where they have been alternatively placed as the sister group of the Otophysi and of the Clupeiformes. In this context, we investigated the phylogeny of the Gonorynchiformes using whole mitogenome sequences from 40 species (six being newly determined for this study). Our taxonomic sampling included at least one species of each gonorynchiform genus and of each other major otocephalan lineage. Unambiguously aligned, concatenated mitogenomic sequences (excluding the ND6 gene and control region) were divided into five partitions (1st, 2nd, and 3rd codon positions, tRNA genes, and rRNA genes) and partitioned Bayesian analyses were conducted. The resultant phylogenetic trees were fully resolved, with most of the nodes well supported by the high posterior probabilities. As expected, the Otocephala were recovered as monophyletic. Within this group, the mitogenome data supported the monophyly of Alepocephaloidei, Gonorynchiformes, Otophysi, and Clupeiformes. The Gonorynchiformes and the Otophysi formed a sister group, rending the Ostariophysi monophyletic. This result conflicts with previous mitogenomic phylogenetic studies, in which a sister relationship was found between Clupeiformes and Gonorynchiformes. We discussed the possible causes of this incongruence. Within the Gonorynchiformes, the following original topology was found: (Gonorynchus (Chanos (Phractolaemus (Cromeria (Grasseichthys (Kneria, Parakneria)))))). We confirmed that the paedomorphic species Cromeria nilotica and Grasseichthys gabonensis belong to the family Kneriidae; however, the two species together did not form a monophyletic group. This result challenges the value of reductive or absent characters as synapomorphies in this group.

Animals↗

Assessment of allelic variation in the ccrAB locus in methicillin-resistant Staphylococcus aureus clones.

OBJECTIVES: To evaluate the allelic variability of ccrA and ccrB among methicillin-resistant Staphylococcus aureus (MRSA) strains belonging to different genetic lineages and carrying different SCCmec types. METHODS: Internal fragments from both genes were obtained by PCR amplification with pairs of degenerate primers and the nucleotide sequence was determined for both strands. Nucleotide sequences were aligned and the phylogenetic trees were reconstructed. RESULTS: The homology scores obtained confirmed the very close relationships among ccrAB alleles associated with each SCCmec type, which further validate this locus as a good target for SCCmec typing strategies. Using this strategy, particularly for the ccrB gene, SCCmec types with the same ccrAB allotype, as detected by PCR, could be discriminated and some strains with non-typeable ccrAB loci were resolved. CONCLUSIONS: We propose that sequencing an internal fragment of ccrB is a strategy for SCCmec typing that may be easily incorporated into other sequence-based MRSA typing strategies, such as multi-locus sequence typing and spa typing.

Alleles↗

Molecular phylogeny and evolution of unique mud-using territorial behavior in ocypodid crabs (Crustacea: Brachyura: Ocypodidae).

Among crabs of the family Ocypodidae, Ilyoplax has been known to exhibit unique mud-using territorial behavior against neighbors, including neighbor burrow plugging, barricade building, and fence building. To assess the evolution of current behavioral forms observed in Ilyoplax, 1,416-bp nucleotide sequences from the mitochondrial 12S rRNA to 16S rRNA genes of 20 species, representing four recognized subfamilies of Ocypodidae, were analyzed. The resultant phylogenetic tree revealed the subfamily Dotillinae, including Ilyoplax, to be monophyletic, with a sister group relationship with subfamily Camptandriinae. These two subfamilies were branched after Ocypodinae, with Macrophthalminae being most basal. Species of Ilyoplax fell into three different Dotillinae lineages, indicating the genus to be polyphyletic. Crabs in two of the three lineages showed differential geographic distribution and body size. Phylogenetic analyses of behavioral characters demonstrated that mud-using techniques had evolved multiple times and sequentially. From their behavioral similarity and evolutionary occurrence, fence building is hypothesized to have evolved from barricade building, and the latter, from burrow plugging. This scenario also appeared reasonable with respect to behavioral trends observed in the field. The evolution of such territorial behavior is considered to be associated with ecological conditions such as burrow fidelity and substrate condition.

Animals↗

The cpcB-cpcA locus as a tool for the genetic characterization of the genus Arthrospira (Cyanobacteria): evidence for horizontal transfer.

To investigate the genetic diversity of the genus Arthrospira and to compare it with other cyanobacteria, sequences of 670 nt from the phycocyanin operon were determined for 23 natural, cultivated or commercial strains of Arthrospira and compared with sequences from 20 other non-Arthrospira cyanobacterial strains. The sequenced DNA fragment comprises the last 255 nt of cpcB, the cpcB-cpcA spacer and the first 304 nt of cpcA. The resulting phylogenetic tree confirms that the genus Arthrospira is not related to Spirulina. So far, cpcB-cpcA data suggest that the closest relative of Arthrospira is Planktothrix. Based on this locus, the genus Arthrospira consists of three genetically clustered lineages. However, the distribution of nucleotide substitutions indicates that these three lineages are not the result of a simple cladogenesis characterized by the accumulation of independent substitutions. Instead, the observed clustering is the result of horizontal transfers of blocks of sequences. Analysis of the distribution of substitutions in the sequenced fragment indicates a point of intragenic recombination close to the stop codon of cpcB. The capacity of exchange of genetic material among strains probably explains why morphology and geographical origin do not correlate with the cpcB-cpcA clusters. Nevertheless, this study shows for the first time that the genus Arthrospira, represented here by cultivated and wild specimens, is clearly monophyletic. Moreover, the cpcB-cpcA DNA fragment, comprising both highly and moderately variable regions, allows (1) a strict differentiation of the taxon Arthrospira from other cyanobacteria (using the coding regions only) and (2) the study of relationships inside Arthrospira (using both the coding and non-coding regions).

Bacterial Proteins↗

Mitochondrial D-loop sequence variation among Italian horse breeds.

The genetic variability of the mitochondrial D-loop DNA sequence in seven horse breeds bred in Italy (Giara, Haflinger, Italian trotter, Lipizzan, Maremmano, Thoroughbred and Sarcidano) was analysed. Five unrelated horses were chosen in each breed and twenty-two haplotypes were identified. The sequences obtained were aligned and compared with a reference sequence and with 27 mtDNA D-loop sequences selected in the GenBank database, representing Spanish, Portuguese, North African, wild horses and an Equus asinus sequence as the outgroup. Kimura two-parameter distances were calculated and a cluster analysis using the Neighbour-joining method was performed to obtain phylogenetic trees among breeds bred in Italy and among Italian and foreign breeds. The cluster analysis indicates that all the breeds but Giara are divided in the two trees, and no clear relationships were revealed between Italian populations and the other breeds. These results could be interpreted as showing the mixed origin of breeds bred in Italy and probably indicate the presence of many ancient maternal lineages with high diversity in mtDNA sequences.

Animals↗

Genetic structure and gene flow in Gran Chaco populations of Argentina: evidence from Y-chromosome markers.

The Gran Chaco region of central South America has been settled by humans for only the last 4,000-5,000 years. To investigate population structure and variation in this region's indigenous population, we scored males from tribes of the Argentinean part of the Gran Chaco (Pilagá, Wichí, and Toba, representing two major language groups, the Mataco and Guaycurú) for a number of Y-chromosome polymorphisms. The markers included eight microsatellites (DYS19, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, and DYS439) and the unique native American single nucleotide polymorphism, DYS199. Sixty males (77%) from the total sample carried the DYS199T chromosome, and these were the focus of the present analysis. Unlike most other native Americans, Gran Chaco males show a moderate level of diversity at the DYS19 locus but still less than that seen in non-native Americans. The FST value for Y-chromosome markers in Gran Chaco was 0.107, a value that is more than double that found for mtDNA haplogroups in the same tribes but is not particularly high compared with other Y-chromosome studies. Phylogenetic trees based on all eight microsatellites showed relatively poor correlation of the tribes with either geography or language, and this is possibly explained by their ecology. They are seasonal hunters living in small bands, and under such circumstances drift will be a powerful evolutionary force. An UPGMA tree based on five microsatellites (DYS19, DYS390, DYS391, DYS392, and DYS393), however, showed a more positive relationship, suggesting that DYS437, DYS438, and DYS439 may behave differently from the other microsatellites. No association was found between maternal and paternal lineage distributions. The time to the most recent common ancestor of the DYS199T chromosome is calculated to lie between 13,000 and 26,000 years. This range is consistent with estimates from other Y-chromosome studies as well as with estimates from mtDNA and the archeology of the colonization of South America. We conclude that the male lineages present in the contemporary Gran Chaco population reflect the level of diversity found in South America and that the region's male founders did not carry a restricted gene pool.

Argentina↗

Methicillin-resistant Staphylococcus aureus (MRSA) infection in hospitalized patients is dominated by community-acquired strains: genomic epidemiological evidence.

OBJECTIVE: This study aimed to systematically investigate the molecular epidemiological characteristics of methicillin-resistant Staphylococcus aureus (MRSA) in Ningxia hospitals, to elucidate their genetic evolutionary relationships, and to delineate the genomic and phenotypic profiles of the dominant lineages. METHODS: Clinical isolates of MRSA strains collected between 01/01/2024 and 30/06/2024 were analyzed, employing second-generation gene sequencing technology, combined with MLST and SCCmec typing, along with evaluation of drug resistance and virulence genes. A phylogenetic tree was constructed to analyze strain homology. RESULTS: A total of 74 non-duplicate Staphylococcus aureus strains (67 MRSA and 7 MSSA) were collected. The most common clonal strain was ST59-IVa, accounting for 46.27%. This strain exhibited a high prevalence of resistance genes mecA and blaZ, at 91.04%. All five ST22-IVa strains were found to lack mecA and erm genes but showed β-lactam resistance, while possessing both lukS/F-PV (PVL) and tsst-1 virulence genes, indicating a significant toxicity risk. Genetic evolution analysis revealed that ST3355, ST4513, and ST59 were closely related, all belonging to SCCmec types IVa; the other ST types exhibited mutations at various loci, with ST5 as the central node, resulting in a wider array of ST and SCCmec typing. CONCLUSION: The ST59-IVa clone is the predominant MRSA strain in Ningxia hospitals, exhibiting multidrug resistance and virulence gene profiles consistent with national trends. However, the emergence of hypervirulent ST22-IVa strains with atypical resistance mechanisms warrants increased vigilance. We recommend enhancing the rational use of antibiotics in hospitals and implementing molecular surveillance for these highly virulent strains.

Methicillin-Resistant Staphylococcus aureus↗

Phylogenetic performance of mitochondrial protein-coding genes in resolving relationships among vertebrates.

A large number of studies in evolutionary biology utilize phylogenetic information obtained from mitochondrial DNA. Researchers place trust in this molecule and expect it generally to be a reliable marker for addressing questions ranging from population genetics to phylogenies among distantly related lineages. Yet, regardless of the phylogenetic method and weighting treatment, individual mitochondrial genes might potentially produce misleading evolutionary inferences and hence might not constitute an adequate representation neither of the entire mitochondrial genome nor of the evolutionary history of the organisms from which they are derived. We investigated the performance of all mitochondrial protein-coding genes to recover two expected phylogenies of tetrapods and mammals. According to these tests, mitochondrial protein-coding genes can be roughly classified into three groups of good (ND4, ND5, ND2, cytb, and COI), medium (COII, COIII, ND1, and ND6), and poor (ATPase 6, ND3, ATPase 8, and ND4L) phylogenetic performers in recovering these expected trees among phylogenetically distant relatives. How general our findings are is unclear. Simple length differences and rate differences between these genes cannot account for their different phylogenetic performance. The phylogenetic performance of these mitochondrial genes might depend on various factors that play a role in determining the probability of discovering the correct phylogeny such as the density of lineage creation events in time, the phylogenetic "depth" of the question, lineage-specific rate heterogeneity, and the completeness of taxa representation.

Adenosine Triphosphatases↗

Heterogeneity of nucleotide frequencies among evolutionary lineages and phylogenetic inference.

A major assumption of many molecular phylogenetic methods is the homogeneity of nucleotide frequencies among taxa, which refers to the equality of the nucleotide frequency bias among species. Changes in nucleotide frequency among different lineages in a data set are thought to lead to erroneous phylogenetic inference because unrelated clades may appear similar because of evolutionarily unrelated similarities in nucleotide frequencies. We tested the effects of the heterogeneity of nucleotide frequency bias on phylogenetic inference, along with the interaction between this heterogeneity and stratified taxon sampling, by means of computer simulations using evolutionary parameters derived from genomic databases. We found that the phylogenetic trees inferred from data sets simulated under realistic, observed levels of heterogeneity for mammalian genes were reconstructed with accuracy comparable to those simulated with homogeneous nucleotide frequencies; the results hold for Neighbor-Joining, minimum evolution, maximum parsimony, and maximum-likelihood methods. The LogDet distance method, specifically designed to deal with heterogeneous nucleotide frequencies, does not perform better than distance methods that assume substitution pattern homogeneity among sequences. In these specific simulation conditions, we did not find a significant interaction between phylogenetic accuracy and substitution pattern heterogeneity among lineages, even when the taxon sampling is increased.

Algorithms↗

Phylogeny of Cryptocercus species (Blattodea: Cryptocercidae) inferred from nuclear ribosomal DNA.

The wood-feeding cockroaches of the genus Cryptocercus occur in temperate forests. Of the seven known species, five occur in the United States and two in Eurasia. Until 1997, all populations in the United States were considered a single species. Populations in the western United States were elevated to a species status based on variation in DNA sequence and morphology. In 1999, three new species were described from the eastern United States based on variation in chromosome number and mitochondrial DNA, bringing the number of species in the United States to five. The objective of this study was to determine if the DNA sequence of nuclear rRNA also signals the existence of four species in the eastern United States and to compare the inferred relationships with those proposed based on mitochondrial sequences. We obtained the DNA sequence from a portion of the 5.8S and 28S rRNA genes and the entire ITS2 region from 38 individuals and 30 additional clones to assess intraindividual, intraspecific, and interspecific variation. We found extensive sequence variation among the various species and little or no intraindividual and intraspecific variation. Phylogenetic analysis indicated the existence of monophyletic lineages among the eastern United States samples, which largely corresponded to the four species previously described. The inferred relationships were well-supported by bootstrap analysis and decay indices. Although the nuclear rRNA sequences resulted in a coherent phylogenetic tree, the ITS2 region contained many insertions and deletions, which may introduce homoplasy and ambiguity in alignment as more taxa are added to the data set.

Animals↗

Mitogenomic evidence for the monophyly of elopomorph fishes (Teleostei) and the evolutionary origin of the leptocephalus larva.

The monophyly of Elopomorpha (eels and their relatives) has long been one of the most problematic issues in systematic ichthyology. Since established the Elopomorpha based on the existence of the leaf-like larval form, termed a leptocephalus, no one has corroborated their monophyly using character matrices derived from both morphological and molecular data during the last 30 years. We investigated their monophyly and interrelationships at the ordinal level using complete mitochondrial genomic (mitogenomic) data from 33 purposefully chosen species (data for nine species being newly determined during the study) that fully represent the major teleostean and elopomorph lineages. Partitioned Bayesian analyses were conducted with the two data sets that comprised concatenated nucleotide sequences from 12 protein-coding genes (with and without third codon positions), 22 transfer RNA genes, and two ribosomal RNA genes. The resultant trees were well resolved and largely congruent, with most internal branches being supported by high statistical values. Mitogenomic data strongly supported the monophyly of Elopomorpha, indicating the validity of the leptocephalus as an elopomorph synapomorphy. The order Elopiformes occupied the most basal position in the elopomorph phylogeny, with the Albuliformes and a clade comprising the Anguilliformes and the Saccopharyngiformes forming a sister group. The most parsimonious reconstruction of the three previously recognized, distinct larval types of elopomorphs onto the molecular phylogeny revealed that one of the types (fork-tailed type) had originated as the common ancestor of the Elopomorpha, the other two (filament-tailed and round-tailed types) having diversified separately in two more derived major clades.

Animals↗

Mitochondrial DNA, morphology, and the phylogenetic relationships of Antarctic icefishes (Notothenioidei: Channichthyidae).

The Channichthyidae is a lineage of 16 species in the Notothenioidei, a clade of fishes that dominate Antarctic near-shore marine ecosystems with respect to both diversity and biomass. Among four published studies investigating channichthyid phylogeny, no two have produced the same tree topology, and no published study has investigated the degree of phylogenetic incongruence between existing molecular and morphological datasets. In this investigation we present an analysis of channichthyid phylogeny using complete gene sequences from two mitochondrial genes (ND2 and 16S) sampled from all recognized species in the clade. In addition, we have scored all 58 unique morphological characters used in three previous analyses of channichthyid phylogenetic relationships. Data partitions were analyzed separately to assess the amount of phylogenetic resolution provided by each dataset, and phylogenetic incongruence among data partitions was investigated using incongruence length difference (ILD) tests. We utilized a parsimony-based version of the Shimodaira-Hasegawa test to determine if alternative tree topologies are significantly different from trees resulting from maximum parsimony analysis of the combined partition dataset. Our results demonstrate that the greatest phylogenetic resolution is achieved when all molecular and morphological data partitions are combined into a single maximum parsimony analysis. Also, marginal to insignificant incongruence was detected among data partitions using the ILD. Maximum parsimony analysis of all data partitions combined results in a single tree, and is a unique hypothesis of phylogenetic relationships in the Channichthyidae. In particular, this hypothesis resolves the phylogenetic relationships of at least two species (Channichthys rhinoceratus and Chaenocephalus aceratus), for which there was no consensus among the previous phylogenetic hypotheses. The combined data partition dataset provides substantial statistical power to discriminate among alternative hypotheses of channichthyid relationships. These findings suggest the optimal strategy for investigating the phylogenetic relationships of channichthyids is one that uses all available phylogenetic data in analyses of combined data partitions.

Animals↗

Evolutionary instability of the major histocompatibility complex class I loci in New World primates.

Homologues of the human major histocompatibility complex (MHC) HLA-A, -B, -E, -F, and -G loci are present in all the Catarrhini (Old World primates, apes, and humans), and some of their allelic lineages have survived several speciation events. Analysis of 26 MHC class I cDNAs from seven different genera of New World primates revealed that the Callitrichinae (tamarins and marmosets) are an exception to these rules of MHC stability. In gene trees of primate MHC class I genes, sequences from the Callitrichinae cluster in a genus-specific fashion, whereas in the other genera of New World primates, as in the Catarrhini, they cluster in a transgeneric way. The genus-specific clustering of the Callitrichinae cDNAs indicates that there is no orthology between MHC class I loci in genera of this phyletic group. Additionally, the Callitrichinae genera exhibit limited variability of their MHC class I genes, in contrast to the high variability displayed by all other primates. Each Callitrichinae genus, therefore, expresses its own set of MHC class I genes, suggesting that an unusually high rate of turnover of loci occurs in this subfamily. The limited variability of MHC class I genes in the Callitrichinae is likely the result of the recent origin of these loci.

Animals↗

Evidence from beta-tubulin phylogeny that microsporidia evolved from within the fungi.

Microsporidia are obligate intracellular parasites that were thought to be an ancient eukaryotic lineage based on molecular phylogenies using ribosomal RNA and translation elongation factors. However, this ancient origin of microsporidia has been contested recently, as several other molecular phylogenies suggest that microsporidia are closely related to fungi. Most of the protein trees that place microsporidia with fungi are not well sampled, however, and it is impossible to resolve whether microsporidia evolved from a fungus or from a protistan relative of fungi. We have sequenced beta-tubulins from 3 microsporidia, 4 chytrid fungi, and 12 zygomycete fungi, expanding the representation of beta-tubulin to include all four fungal divisions and a wide diversity of microsporidia. In phylogenetic trees including these new sequences, the overall topology of the fungal beta-tubulins generally matched the expected relationships among the four fungal divisions, although the zygomycetes were polyphyletic in some analyses. The microsporidia consistently fell within this fungal diversification, and not as a sister group to fungi. Overall, beta-tubulin phylogeny suggests that microsporidia evolved from a fungus sometime after the divergence of chytrids. We also found that chytrid alpha- and beta-tubulins are much less divergent than are tubulins from other fungi or microsporidia. In trees in which the only fungal representatives were the chytrids, microsporidia still branched with fungi (i.e., with chytrids), suggesting that the affiliation between microsporidian and fungal tubulins is not an artifact of long-branch attraction.

Animals↗

Alpha-proteobacterial relationship of apicomplexan lactate and malate dehydrogenases.

We have cloned and sequenced a lactate dehydrogenase (LDH) gene from Cryptosporidium parvum (CpLDH1). With this addition, and that of four recently deposited alpha-proteobacterial malate dehydrogenase (MDH) genes, the phylogenetic relationships among apicomplexan LDH and bacterial MDH were re-examined. Consistent with previous studies, our maximum likelihood (ML) analysis using the quartet-puzzling method divided 105 LDH/MDH enzymes into five clades, and confirmed that mitochondrial MDH is a sister clade to those of y-proteobacteria, rather than to alpha-proteobacteria. In addition, a Cryptosporidium parvum MDH (CpMDH1) was identified from the ongoing Cryptosporidium genome project that appears to belong to a distinct clade (III) comprised of 22 sequences from one archaebacterium, numerous eubacteria, and several apicomplexans. Using the ML puzzling test and bootstrapping analysis with protein distance and parsimony methods, the resulting trees not only robustly confirmed the alpha-proteobacterial relationship of apicomplexan LDH/MDH, but also supported a monophyletic relationship of CpLDH1 with CpMDHI. These data suggest that, unlike most other eukaryotes, the Apicomplexa may be one of the few lineages retaining an alpha-proteobacterial-type MDH that could have been acquired from an ancestral alpha-proteobacterium through primary endosymbiosis giving rise to the mitochondria, or through an unknown lateral gene transfer (LGT) event.

Alphaproteobacteria↗

On the mtDNA restriction patterns variation of the Iberian wood mouse (Apodemus sylvaticus). Comparison with other west Mediterranean populations.

In previous studies, the presence of three main lineages of wood mice mtDNA was described in western Europe: a first one distributed from the Pyrenees to Scandinavia. a Thyrrenian one occurring in peninsular Italy, Elba, Corsica and Sardinia and a third one restricted to Sicily and Marettimo. Do the Iberian wood mice belong to one of these lineages? In order to answer this question, animals were trapped all over the Iberian peninsula (11 sites) as well as in three of the Balearic islands. Comparisons with specimens from the above mentioned lineages were made. From 158 animals trapped in 30 sites, 78 different mtDNA restriction patterns were obtained and compared using the NEI and LI index ((Nei M and Li WH, (1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA 76: 5269-5273) of nucleotide divergence. A neighbour-joining tree, with a specimen of Apodemus flavicollis as outgroup, was then computed from the similarity matrix. Apodemus flavicollis is well separated from all the A. sylvaticus and all the Iberian restriction patterns are clustered with those of continental France, showing a great similarity level between the Iberian and north-west European animals. This group is well separated from the Sicilian and from the Tyrrhenian ones. These results suggest that the Pyrenees are not a biogeographic barrier for the wood mouse and that the postglacial recolonisation of western Europe by that species has its origin in populations which, during the latest Ice age, were living in refuges situated in southern France or in the Iberian peninsula. Because the Balearic restriction patterns are clustered in a lineage joining Iberian pattern at a low divergence level, we suggest that their origin is continental Spain or southern France.

Animals↗

NifH and NifD sequences of heliobacteria: a new lineage in the nitrogenase phylogeny.

We determined almost complete nifH and nifD genes from representatives of all recognized genera of heliobacteria, the strictly anaerobic phototrophs belonging to the low GC gram-positive bacteria. The heliobacterial sequences formed a highly supported monophyletic group that is clearly distinct from any known diazotrophs, in both NifH and NifD trees. According to the classification of nitrogenase genes in four major clusters, the clade of heliobacterial sequences belonged to cluster I and did not cluster with any of the Clostridium (cluster III) or Paenibacillus (cluster I) species, the close neighbors of heliobacteria based on the 16S rRNA phylogeny. One partial anfH or alternative nitrogenase sequence was detected from Heliobacterium gestii. Although Heliophilum fasciatum is known to fix nitrogen based on the acetylene reduction test, nifH and/or nifD genes were not detected by either the PCR amplification or Southern hybridization methods.

Bacillus↗