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Lara, Enrique

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Lara, Enrique
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https://libra.unine.ch/handle/123456789/360

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  • Publication
    Accès libre
    Assessing the responses of Sphagnum micro-eukaryotes to climate changes using high throughput sequencing
    (2020-9-18)
    Reczuga, Monika 
    ;
    Seppey, Christophe Victor William
    ;
    Mulot, Matthieu 
    ;
    Jassey, Vincent E.J.
    ;
    Buttler, Alexandre
    ;
    Slowinska, Sandra
    ;
    Slowinski, Michal
    ;
    Lara, Enrique 
    ;
    Lamentowicz, Mariusz
    ;
    Mitchell, Edward 
    Current projections suggest that climate warming will be accompanied by more frequent and severe drought events. Peatlands store ca. one third of the world’s soil organic carbon. Warming and drought may cause peatlands to become carbon sources through stimulation of microbial activity increasing ecosystem respiration, with positive feedback effect on global warming. Micro-eukaryotes play a key role in the carbon cycle through food web interactions and therefore, alterations in their community structure and diversity may affect ecosystem functioning and could reflect these changes. We assessed the diversity and community composition of Sphagnum-associated eukaryotic microorganisms inhabiting peatlands and their response to experimental drought and warming using high throughput sequencing of environmental DNA. Under drier conditions, micro-eukaryotic diversity decreased, the relative abundance of autotrophs increased and that of osmotrophs (including Fungi and Peronosporomycetes) decreased. Furthermore, we identified climate change indicators that could be used as early indicators of change in peatland microbial communities and ecosystem functioning. The changes we observed indicate a shift towards a more “terrestrial” community in response to drought, in line with observed changes in the functioning of the ecosystem.
  • Publication
    Accès libre
    rRNA Phylogeny of Arcellinida (Amoebozoa) Reveals that the Largest Arcellinid Genus, Difflugia Leclerc 1815, is not Monophyletic
    (2012)
    Gomaa, Fatma
    ;
    Todorov, Milcho
    ;
    Heger, Thierry J.
    ;
    Mitchell, Edward 
    ;
    Lara, Enrique 
    The systematics of lobose testate amoebae (Arcellinida), a diverse group of shelled free-living unicellular eukaryotes, is still mostly based on morphological criteria such as shell shape and composition. Few molecular phylogenetic studies have been performed on these organisms to date, and their phylogeny suffers from typical under-sampling artefacts, resulting in a still mostly unresolved tree. In order to clarify the phylogenetic relationships among arcellinid testate amoebae at the inter-generic and inter-specific level, and to evaluate the validity of the criteria used for taxonomy, we amplified and sequenced the SSU rRNA gene of nine taxa - Difflugia bacillariarum, D. hiraethogii, D. acuminata, D. lanceolata, D. achlora, Bullinularia gracilis, Netzelia oviformis, Physochila griseola and Cryptodifflugia oviformis. Our results, combined with existing data demonstrate the following: 1) Most arcellinids are divided into two major clades, 2) the genus Difflugia is not monophyletic, and the genera Netzelia and Arcella are closely related, and 3) Cryptodifflugia branches at the base of the Arcellinida clade. These results contradict the traditional taxonomy based on shell composition, and emphasize the importance of general shell shape in the taxonomy of arcellinid testate amoebae.
  • Publication
    Accès libre
    COI Barcoding of Nebelid Testate Amoebae (Amoebozoa: Arcellinida): Extensive Cryptic Diversity and Redefinition of the Hyalospheniidae Schultze
    (2012)
    Kosakyan, Anush 
    ;
    Heger, Thierry J.
    ;
    Leander, Brian S.
    ;
    Todorov, Milcho
    ;
    Mitchell, Edward 
    ;
    Lara, Enrique 
    We used Cytochrome Oxidase Subunit 1 (COI) to assess the phylogenetic relationships and taxonomy of Nebela sensu stricto and similar taxa (Nebela group, Arcellinida) in order to clarify the taxonomic validity of morphological characters. The COI data not only successfully separated all studied morphospecies but also revealed the existence of several potential cryptic species. The taxonomic implications of the results are: (1) Genus Nebela is paraphyletic and will need to be split into at least two monophyletic assemblages when taxon sampling is further expanded. (2) Genus Quadrulella, one of the few arcellinid genera building its shell from self-secreted siliceous elements, and the mixotrophic Hyalosphenia papilio branch within the Nebela group in agreement with the general morphology of their shell and the presence of an organic rim around the aperture (synapomorphy for Hyalospheniidae). We thus synonymise Hyalospheniidae and Nebelidae. Hyalospheniidae takes precedence and now includes Hyalosphenia, Quadrulella (previously in the Lesquereusiidae) and all Nebelidae with the exception of Argynnia and Physochila. Leptochlamys is Arcellinida incertae sedis. We describe a new genus Padaungiella Lara et Todorov and a new species Nebela meisterfeldi n. sp. Heger et Mitchell and revise the taxonomic position (and rank) of several taxa. These results show that the traditional morphology-based taxonomy underestimates the diversity within the Nebela group, and that phylogenetic relationships are best inferred from shell shape rather than from the material used to build the shell.
  • Publication
    Accès libre
    Comparing Potential COI and SSU rDNA Barcodes for Assessing the Diversity and Phylogenetic Relationships of Cyphoderiid Testate Amoebae (Rhizaria: Euglyphida)
    (2011)
    Heger, Thierry J.
    ;
    Pawlowski, Jan
    ;
    Lara, Enrique 
    ;
    Leander, Brian S.
    ;
    Todorov, Milcho
    ;
    Golemansky, Vassil
    ;
    Mitchell, Edward 
    The mitochondrial Cytochrome Oxidase Subunit 1 gene (COI) has been promoted as an ideal “DNA barcode” for animal species and other groups of eukaryotes. However, the utility of the COI marker for species level discrimination and for phylogenetic analyses has yet to be tested within the Rhizaria. Accordingly, we analysed mitochondrial COI gene sequences and nuclear small subunit rDNA (SSU) sequences from several morphospecies of euglyphid testate amoebae (Cercozoa, Rhizaria) in order to evaluate the utility of these DNA markers for species discrimination and phylogenetic reconstructions. Sequences were obtained from eleven populations belonging to six Cyphoderia morphospecies that were isolated from field samples in North America and Europe. Mean inter-population COI sequence dissimilarities were on average 2.9 times greater than in the SSU, while the intra-population sequence dissimilarities were higher in the SSU (0-0.95%) than in the COI (0%); this suggests that the COI fragment is valuable for discriminating Cyphoderiidae isolates. Our study also demonstrated that COI sequences are useful for inferring phylogenetic relationships among Cyphoderiidae isolates. COI and SSU tree topologies were very similar even though the COI fragment used in these analyses (500 bp) was much shorter than the SSU sequences (1600 bp). Altogether, these results demonstrate the utility of the COI as a potential taxonomic DNA barcode for assessing cyphoderiid species diversity and for inferring phylogenetic relationships within the group.
  • Publication
    Accès libre
    Highly Diverse and Seasonally Dynamic Protist Community in a Pristine Peat Bog
    (2011)
    Lara, Enrique 
    ;
    Mitchell, Edward 
    ;
    Moreira, David
    ;
    López-García, Purificación
    Culture-independent molecular methods based on the amplification, cloning and sequencing of small-subunit ribosomal RNA genes (SSU rDNAs) are powerful tools to study the diversity of microorganisms. Despite so, the eukaryotic microbial diversity of many ecosystems, including peatlands has not yet received much attention. We analysed the eukaryotic diversity by molecular surveys in water from the centre of a pristine Sphagnum-dominated peatland in the Jura Mountains of Switzerland during a complete seasonal cycle. The clone libraries constructed from five different temporal samplings revealed a high diversity of protists with representatives of all major eukaryotic phyla. In addition, four sequence types could not be assigned to any known high-level eukaryotic taxon but branched together with a rather good statistic support, raising the possibility of a novel, deep branching eukaryotic clade. The analysis of seasonal patterns of phylotypes showed a clear change in the eukaryotic communities between the warm period (late spring and summer) and the cold period (autumn and winter). Chrysophytes dominated the samples in the cold period while testate amoebae (Arcellinida and Euglyphida) and a few other groups peaked in summer. A few phylotypes (such as a cryptomonad and a perkinsid) were abundant at given sampling times and then almost disappeared, suggesting bloom-like dynamics.
  • Publication
    Accès libre
    Molecular phylogeny of euglyphid testate amoebae (Cercozoa: Euglyphida) suggests transitions between marine supralittoral and freshwater/terrestrial environments are infrequent
    (2010)
    Heger, Thierry J.
    ;
    Mitchell, Edward 
    ;
    Todorov, Milcho
    ;
    Golemansky, Vassil
    ;
    Lara, Enrique 
    ;
    Leander, Brian S.
    ;
    Pawlowski, Jan
    Marine and freshwater ecosystems are fundamentally different regarding many biotic and abiotic factors. The physiological adaptations required for an organism to pass the salinity barrier are considerable. Many eukaryotic lineages are restricted to either freshwater or marine environments. Molecular phylogenetic analyses generally demonstrate that freshwater species and marine species segregate into different sub-clades, indicating that transitions between these two environments occur only rarely in the course of evolution. It is, however, unclear if the transitions between freshwater and environments characterized by highly variable salinities, such as the marine supralittoral zone, are also infrequent. Here, we use testate amoebae within the Euglyphida to assess the phylogenetic interrelationships between marine supralittoral and freshwater taxa. Euglyphid testate amoebae are mainly present in freshwater habitats but also occur in marine supralittoral environments. Accordingly, we generated and analyzed partial SSU rRNA gene sequences from 49 new marine/supralittoral and freshwater Cyphoderiidae sequences, 20 sequences of the Paulinellidae, Trinematidae, Assulinidae, and Euglyphidae families as well as 21 GenBank sequences of unidentified taxa derived from environmental PCR surveys. Both the molecular and morphological data suggest that the diversity of Cyphoderiidae is strongly underestimated. The results of our phylogenetic analyses demonstrated that marine supralittoral and freshwater euglyphid testate amoeba species are segregated into distinct sub-clades, suggesting that transitions between these two habitats occurred only infrequently.
  • Publication
    Accès libre
    COI gene and ecological data suggest size-dependent high dispersal and low intra-specific diversity in free-living terrestrial protists (Euglyphida: Assulina)
    (2010)
    Lara, Enrique 
    ;
    Heger, Thierry J.
    ;
    Scheihing, Rodrigo
    ;
    Mitchell, Edward 
    Aim  Propagule size and ecological requirements are believed to be major factors influencing the passive dispersal of free-living terrestrial protists. We compared the colonization potential of three closely related testate amoeba species (Assulina muscorum, A. seminulum, A. scandinavica, ranging from 40 to 100 μm in length).
    Location  Europe.
    Methods  We collected individual Assulina species cells from Sphagnum peatlands across Europe. We sequenced a 550-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI) to estimate the within-species variability, as a proxy for gene flow. We reviewed existing ecological and palaeoecological data to assess the ecological tolerance of Assulina species and how rapidly they colonized developing peatlands.
    Results  We obtained COI sequences for 30 individuals of A. seminulum from eleven sites, for 39 of A. muscorum from six sites, and for six of A. scandinavica from two sites. We observed three haplotypes for A. seminulum and two for A. muscorum, often co-existing in the same sites. The sequences of A. scandinavica from the two sites were identical. Assulina muscorum and A. seminulum haplotypes varied by only 1–2 nucleotides, resulting in >99.5% similarity. Genetic diversity within A. seminulum was higher than that within A. muscorum. Ecological and palaeoecological records showed that A. muscorum was much more frequent and abundant than A. seminulum, and had a somewhat broader ecological tolerance for pH, moisture and water-table depth. Assulina muscorum always appeared early during the developmental history of peatlands, either before or simultaneously with A. seminulum.
    Main conclusions  The lack of genetic structure observed with a variable marker such as COI suggests high gene flow between the sites and thus rapid transport (at an evolutionary scale) over large distances, in agreement with the palaeoecological records. Thus, geographical distance alone does not seem to prevent the dispersal of testate amoebae, at least within Europe. Nevertheless, genetic diversity was significantly lower within A. muscorum than within A. seminulum, suggesting that its smaller size and abundance and/or broader ecological tolerance influence its effective dispersal capacity. These results are in agreement with the often earlier colonization of peatlands by A. muscorum and its broader ecological tolerance.
  • Publication
    Accès libre
    Ribosomal RNA Genes Challenge the Monophyly of the Hyalospheniidae (Amoebozoa: Arcellinida)
    (2008)
    Lara, Enrique 
    ;
    Heger, Thierry J.
    ;
    Ekelund, Flemming
    ;
    Lamentowicz, Mariucz
    ;
    Mitchell, Edward 
    To date only five partial and two complete SSU rRNA gene sequences are available for the lobose testate amoebae (Arcellinida). Consequently, the phylogenetic relationships among taxa and the definition of species are still largely dependant on morphological characters of uncertain value, which causes confusion in the phylogeny, taxonomy and the debate on cosmopolitanism of free-living protists. Here we present a SSU rRNA-based phylogeny of the Hyalospheniidae including the most common species. Similar to the filose testate amoebae of the order Euglyphida the most basal clades have a terminal aperture; the ventral position of the pseudostome appears to be a derived character. Family Hyalospheniidae appears paraphyletic and is separated into three clades: (1) Heleopera sphagni, (2) Heleopera rosea and Argynnia dentistoma and (3) the rest of the species from genera Apodera, Hyalosphenia, Porosia and Nebela. Our data support the validity of morphological characters used to define species among the Hyalospheniidae and even suggest that taxa described as varieties may deserve the rank of species (e.g. N. penardiana var. minor). Finally our results suggest that the genera Hyalosphenia and Nebela are paraphyletic, and that Porosia bigibbosa branches inside the main Nebela clade.
  • Publication
    Accès libre
    SSU rRNA reveals a sequential increase in shell complexity among the euglyphid testate amoebae (Rhizaria : Euglyphida)
    (2007)
    Lara, Enrique 
    ;
    Heger, Thierry J
    ;
    Mitchell, Edward 
    ;
    Meisterfeld, Ralf
    ;
    Ekelund, Flemming
    The existing data on the molecular phylogeny of filose testate amoebae from order Euglyphida has revealed contradictions between traditional morphological classification and SSU rRNA phylogeny and, moreover, the position of several important genera remained unknown. We therefore carried out a study aiming to fill several important gaps and better understand the relationships among the main euglyphid testate amoebae and the evolutionary steps that led to the present diversity at a higher level. We obtained new SSU rRNA sequences from five genera and seven species. This new phylogeny obtained shows that (1) the clade formed by species of genera Assulina and Placocista branches unambiguously at the base of the subclade of Euglyphida comprising all members of the family Trinematidae and genus Euglypha, (2) family Trinematidae (Trachelocorythion, Trinema, and Corythion) branches as a sister group to genus Euglypha, (3) three newly sequenced Euglypha species (E. cf. ciliata, E. penardi, and E. compressa) form a new clade within the genus. Since our results show that Assulina and Placocista do not belong to the Euglyphidae (unless the Trinematidae are also included in this family), we propose the creation of a new family named Assulinidae. Consequently, we give a family status to the genera Euglypha and (tentatively) Scutiglypha, which become the new family Euglyphidae. The evolutionary pattern suggested by SSU rRNA phylogeny shows a clear tendency towards increasing morphological complexity of the shell characterised by changes in the symmetry (migration of the aperture to a ventral position and/or compression of the shell) and the appearance of specialised scales at the aperture (in families Trinematidae and Euglyphidae). (C) 2007 Elsevier GmbH. All rights reserved.