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1.
Nature ; 541(7638): 536-540, 2017 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-28092920

RESUMO

The Southern Ocean houses a diverse and productive community of organisms. Unicellular eukaryotic diatoms are the main primary producers in this environment, where photosynthesis is limited by low concentrations of dissolved iron and large seasonal fluctuations in light, temperature and the extent of sea ice. How diatoms have adapted to this extreme environment is largely unknown. Here we present insights into the genome evolution of a cold-adapted diatom from the Southern Ocean, Fragilariopsis cylindrus, based on a comparison with temperate diatoms. We find that approximately 24.7 per cent of the diploid F. cylindrus genome consists of genetic loci with alleles that are highly divergent (15.1 megabases of the total genome size of 61.1 megabases). These divergent alleles were differentially expressed across environmental conditions, including darkness, low iron, freezing, elevated temperature and increased CO2. Alleles with the largest ratio of non-synonymous to synonymous nucleotide substitutions also show the most pronounced condition-dependent expression, suggesting a correlation between diversifying selection and allelic differentiation. Divergent alleles may be involved in adaptation to environmental fluctuations in the Southern Ocean.


Assuntos
Aclimatação/genética , Temperatura Baixa , Diatomáceas/genética , Evolução Molecular , Genoma/genética , Genômica , Alelos , Dióxido de Carbono/metabolismo , Escuridão , Diatomáceas/metabolismo , Congelamento , Perfilação da Expressão Gênica , Deriva Genética , Camada de Gelo , Ferro/metabolismo , Taxa de Mutação , Oceanos e Mares , Filogenia , Recombinação Genética , Transcriptoma/genética
2.
J Phycol ; 56(3): 747-760, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32068264

RESUMO

The haptophyte Phaeocystis antarctica is endemic to the Southern Ocean, where iron supply is sporadic and its availability limits primary production. In iron fertilization experiments, P. antarctica showed a prompt and steady increase in cell abundance compared to heavily silicified diatoms along with enhanced colony formation. Here we utilized a transcriptomic approach to investigate molecular responses to alleviation of iron limitation in P. antarctica. We analyzed the transcriptomic response before and after (14 h, 24 h and 72 h) iron addition to a low-iron acclimated culture. After iron addition, we observed indicators of a quick reorganization of cellular energetics, from carbohydrate catabolism and mitochondrial energy production to anabolism. In addition to typical substitution responses from an iron-economic toward an iron-sufficient state for flavodoxin (ferredoxin) and plastocyanin (cytochrome c6 ), we found other genes utilizing the same strategy involved in nitrogen assimilation and fatty acid desaturation. Our results shed light on a number of adaptive mechanisms that P. antarctica uses under low iron, including the utilization of a Cu-dependent ferric reductase system and indication of mixotrophic growth. The gene expression patterns underpin P. antarctica as a quick responder to iron addition.


Assuntos
Diatomáceas , Haptófitas , Aclimatação , Diatomáceas/genética , Ferro , Fitoplâncton , Transcriptoma
3.
Rapid Commun Mass Spectrom ; 33(2): 193-202, 2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30366355

RESUMO

RATIONALE: High-resolution mass spectrometry (HRMS) with high sample throughput has become an important analytical tool for the analysis of highly complex samples and data processing has become a major challenge for the user community. Evaluating direct-infusion HRMS data without automated tools for batch processing can be a time-consuming step in the analytical pipeline. Therefore, we developed a new browser-based software tool for processing HRMS data. METHODS: The software, named UltraMassExplorer (UME), was written in the R programming language using the shiny library to build the graphical user interface. The performance of the integrated formula library search algorithm was tested using HRMS data derived from analyses of up to 50 extracts of marine dissolved organic matter. RESULTS: The software supports the processing of lists of calibrated masses of neutral, protonated or deprotonated molecules, with masses of up to 700 Da and a mass accuracy <3 ppm. In the performance test, the number of assigned peaks per second increased with the number of submitted peaks and reached a maximum rate of 4745 assigned peaks per second. CONCLUSIONS: UME offers a complete data evaluation pipeline comprising a fast molecular formula assignment algorithm allowing for the swift reanalysis of complete datasets, advanced filter functions and the export of data, metadata and publication-quality graphics. Unique to UME is a fast and interactive connection between data and their visual representation. UME provides a new platform enabling an increased transparency, customization, documentation and comparability of datasets.

4.
Nature ; 499(7457): 209-13, 2013 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-23760476

RESUMO

Coccolithophores have influenced the global climate for over 200 million years. These marine phytoplankton can account for 20 per cent of total carbon fixation in some systems. They form blooms that can occupy hundreds of thousands of square kilometres and are distinguished by their elegantly sculpted calcium carbonate exoskeletons (coccoliths), rendering them visible from space. Although coccolithophores export carbon in the form of organic matter and calcite to the sea floor, they also release CO2 in the calcification process. Hence, they have a complex influence on the carbon cycle, driving either CO2 production or uptake, sequestration and export to the deep ocean. Here we report the first haptophyte reference genome, from the coccolithophore Emiliania huxleyi strain CCMP1516, and sequences from 13 additional isolates. Our analyses reveal a pan genome (core genes plus genes distributed variably between strains) probably supported by an atypical complement of repetitive sequence in the genome. Comparisons across strains demonstrate that E. huxleyi, which has long been considered a single species, harbours extensive genome variability reflected in different metabolic repertoires. Genome variability within this species complex seems to underpin its capacity both to thrive in habitats ranging from the equator to the subarctic and to form large-scale episodic blooms under a wide variety of environmental conditions.


Assuntos
Genoma/genética , Haptófitas/genética , Haptófitas/isolamento & purificação , Fitoplâncton/genética , Calcificação Fisiológica , Cálcio/metabolismo , Anidrases Carbônicas/genética , Anidrases Carbônicas/metabolismo , Ecossistema , Haptófitas/classificação , Haptófitas/metabolismo , Oceanos e Mares , Filogenia , Proteoma/genética , Água do Mar
5.
Glob Chang Biol ; 23(4): 1499-1510, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27718513

RESUMO

Thermal tolerance windows serve as a powerful tool for estimating the vulnerability of marine species and their life stages to increasing temperature means and extremes. However, it remains uncertain to which extent additional drivers, such as ocean acidification, modify organismal responses to temperature. This study investigated the effects of CO2 -driven ocean acidification on embryonic thermal sensitivity and performance in Atlantic cod, Gadus morhua, from the Kattegat. Fertilized eggs were exposed to factorial combinations of two PCO2 conditions (400 µatm vs. 1100 µatm) and five temperature treatments (0, 3, 6, 9 and 12 °C), which allow identifying both lower and upper thermal tolerance thresholds. We quantified hatching success, oxygen consumption (MO2 ) and mitochondrial functioning of embryos as well as larval morphometrics at hatch and the abundance of acid-base-relevant ionocytes on the yolk sac epithelium of newly hatched larvae. Hatching success was high under ambient spawning conditions (3-6 °C), but decreased towards both cold and warm temperature extremes. Elevated PCO2 caused a significant decrease in hatching success, particularly at cold (3 and 0 °C) and warm (12 °C) temperatures. Warming imposed limitations to MO2 and mitochondrial capacities. Elevated PCO2 stimulated MO2 at cold and intermediate temperatures, but exacerbated warming-induced constraints on MO2 , indicating a synergistic interaction with temperature. Mitochondrial functioning was not affected by PCO2 . Increased MO2 in response to elevated PCO2 was paralleled by reduced larval size at hatch. Finally, ionocyte abundance decreased with increasing temperature, but did not differ between PCO2 treatments. Our results demonstrate increased thermal sensitivity of cod embryos under future PCO2 conditions and suggest that acclimation to elevated PCO2 requires reallocation of limited resources at the expense of embryonic growth. We conclude that ocean acidification constrains the thermal performance window of embryos, which has important implication for the susceptibility of cod to projected climate change.


Assuntos
Mudança Climática , Gadus morhua , Aclimatação , Animais , Concentração de Íons de Hidrogênio , Oceanos e Mares , Temperatura
6.
J Phycol ; 51(1): 93-108, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26986261

RESUMO

Macroalgae of the order Laminariales (kelp) are important components of cold-temperate coastal ecosystems. Major factors influencing their distribution are light (including UV radiation) and temperature. Therefore, future global environmental changes potentially will impact their zonation, distribution patterns, and primary productivity. Many physiological studies were performed on UV radiation and temperature stress in kelp but combinatory effects have not been analyzed and so far no study is available on the molecular processes involved in acclimation to these stresses. Therefore, sporophytes of Saccharina latissima were exposed for 2 weeks to 12 combinations of photosynthetically active radiation (PAR), UV radiation and temperature. Subsequently, microarray hybridizations were performed to determine changes in gene expression patterns. Several effects on the transcriptome were observed after exposure experiments. The strongest effect of temperature on gene expression was observed at 2°C. Furthermore, UV radiation had stronger effects on gene expression than high PAR, and caused stronger induction genes correlated with categories such as photosynthetic components and vitamin B6 biosynthesis. Higher temperatures ameliorated the negative effects of UV radiation in S. latissima. Regulation of reactive oxygen species (ROS) scavenging seems to work in a compartment specific way. Gene expression profiles of ROS scavengers indicated a high amount of oxidative stress in response to the 2°C condition as well as to excessive light at 12°C. Interestingly, stress levels that did not lead to physiological alterations already caused by a transcriptomic response.

7.
BMC Genomics ; 15: 789, 2014 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-25216596

RESUMO

BACKGROUND: Hypercapnia and elevated temperatures resulting from climate change may have adverse consequences for many marine organisms. While diverse physiological and ecological effects have been identified, changes in those molecular mechanisms, which shape the physiological phenotype of a species and limit its capacity to compensate, remain poorly understood. Here, we use global gene expression profiling through RNA-Sequencing to study the transcriptional responses to ocean acidification and warming in gills of the boreal spider crab Hyas araneus exposed medium-term (10 weeks) to intermediate (1,120 µatm) and high (1,960 µatm) PCO2 at different temperatures (5°C and 10°C). RESULTS: The analyses reveal shifts in steady state gene expression from control to intermediate and from intermediate to high CO2 exposures. At 5°C acid-base, energy metabolism and stress response related genes were upregulated at intermediate PCO2, whereas high PCO2 induced a relative reduction in expression to levels closer to controls. A similar pattern was found at elevated temperature (10°C). There was a strong coordination between acid-base, metabolic and stress-related processes. Hemolymph parameters at intermediate PCO2 indicate enhanced capacity in acid-base compensation potentially supported by upregulation of a V-ATPase. The likely enhanced energy demand might be met by the upregulation of the electron transport system (ETS), but may lead to increased oxidative stress reflected in upregulated antioxidant defense transcripts. These mechanisms were attenuated by high PCO2, possibly as a result of limited acid-base compensation and metabolic down-regulation. CONCLUSION: Our findings indicate a PCO2 dependent threshold beyond which compensation by acclimation fails progressively. They also indicate a limited ability of this stenoecious crustacean to compensate for the effects of ocean acidification with and without concomitant warming.


Assuntos
Braquiúros/genética , Dióxido de Carbono/toxicidade , Perfilação da Expressão Gênica/métodos , Brânquias/metabolismo , Água do Mar/química , Análise de Sequência de RNA/métodos , Animais , Braquiúros/anatomia & histologia , Transporte de Elétrons , Metabolismo Energético , Regulação da Expressão Gênica , Concentração de Íons de Hidrogênio , Masculino , Estresse Oxidativo , Temperatura
8.
Glob Chang Biol ; 20(10): 3059-67, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24890266

RESUMO

Organisms in all domains, Archaea, Bacteria, and Eukarya will respond to climate change with differential vulnerabilities resulting in shifts in species distribution, coexistence, and interactions. The identification of unifying principles of organism functioning across all domains would facilitate a cause and effect understanding of such changes and their implications for ecosystem shifts. For example, the functional specialization of all organisms in limited temperature ranges leads us to ask for unifying functional reasons. Organisms also specialize in either anoxic or various oxygen ranges, with animals and plants depending on high oxygen levels. Here, we identify thermal ranges, heat limits of growth, and critically low (hypoxic) oxygen concentrations as proxies of tolerance in a meta-analysis of data available for marine organisms, with special reference to domain-specific limits. For an explanation of the patterns and differences observed, we define and quantify a proxy for organismic complexity across species from all domains. Rising complexity causes heat (and hypoxia) tolerances to decrease from Archaea to Bacteria to uni- and then multicellular Eukarya. Within and across domains, taxon-specific tolerance limits likely reflect ultimate evolutionary limits of its species to acclimatization and adaptation. We hypothesize that rising taxon-specific complexities in structure and function constrain organisms to narrower environmental ranges. Low complexity as in Archaea and some Bacteria provide life options in extreme environments. In the warmest oceans, temperature maxima reach and will surpass the permanent limits to the existence of multicellular animals, plants and unicellular phytoplankter. Smaller, less complex unicellular Eukarya, Bacteria, and Archaea will thus benefit and predominate even more in a future, warmer, and hypoxic ocean.


Assuntos
Adaptação Fisiológica , Organismos Aquáticos , Evolução Biológica , Mudança Climática , Ecossistema , Aclimatação/fisiologia , Archaea , Bactérias , Eucariotos , Temperatura Alta , Oceanos e Mares , Oxigênio/metabolismo
9.
BMC Genomics ; 13: 549, 2012 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-23051706

RESUMO

BACKGROUND: Studies of temperature-induced adaptation on the basis of genomic sequence data were mainly done in extremophiles. Although the general hypothesis of an increased molecular flexibility in the cold is widely accepted, the results of thermal adaptation are still difficult to detect at proteomic down to the genomic sequence level. Approaches towards a more detailed picture emerge with the advent of new sequencing technologies. Only small changes in primary protein structure have been shown to modify kinetic and thermal properties of enzymes, but likewise for interspecies comparisons a high genetic identity is still essential to specify common principles. The present study uses comprehensive transcriptomic sequence information to uncover general patterns of thermal adaptation on the RNA as well as protein primary structure. RESULTS: By comparing orthologous sequences of two closely related zoarcid fish inhabiting different latitudinal zones (Antarctica: Pachycara brachycephalum, temperate zone: Zoarces viviparus) we were able to detect significant differences in the codon usage. In the cold-adapted species a lower GC content in the wobble position prevailed for preserved amino acids. We were able to estimate 40-60% coverage of the functions represented within the two compared zoarcid cDNA-libraries on the basis of a reference genome of the phylogenetically closely related fish Gasterosteus aculeatus. A distinct pattern of amino acid substitutions could be identified for the non-synonymous codon exchanges, with a remarkable surplus of serine and reduction of glutamic acid and asparagine for the Antarctic species. CONCLUSION: Based on the differences between orthologous sequences from confamiliar species, distinguished mainly by the temperature regimes of their habitats, we hypothesize that temperature leaves a signature on the composition of biological macromolecules (RNA, proteins) with implications for the transcription and translation level. As the observed pattern of amino acid substitutions only partly support the flexibility hypothesis further evolutionary forces may be effective at the global transcriptome level.


Assuntos
Adaptação Fisiológica/genética , Evolução Molecular , Perciformes/genética , Perciformes/fisiologia , Temperatura , Transcriptoma , Animais , Regiões Antárticas , Códon/genética , Ecossistema , Biblioteca Gênica , Fígado/metabolismo , Miocárdio/metabolismo , Filogenia , Especificidade da Espécie
10.
Am J Physiol Regul Integr Comp Physiol ; 301(5): R1453-66, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21865546

RESUMO

It is widely accepted that adaptation to the extreme cold has evolved at the expense of high thermal sensitivity. However, recent studies have demonstrated significant capacities for warm acclimation in Antarctic fishes. Here, we report on hepatic metabolic reorganization and its putative molecular background in the Antarctic eelpout (Pachycara brachycephalum) during warm acclimation to 5°C over 6 wk. Elevated capacities of cytochrome c oxidase suggest the use of warm acclimation pathways different from those in temperate fish. The capacity of this enzyme rose by 90%, while citrate synthase (CS) activity fell by 20% from the very beginning. The capacity of lipid oxidation by hydroxyacyl-CoA dehydrogenase remained constant, whereas phosphoenolpyruvate carboxykinase as a marker for gluconeogenesis displayed 40% higher activities. These capacities in relation to CS indicate a metabolic shift from lipid to carbohydrate metabolism. The finding was supported by large rearrangements of the related transcriptome, both functional genes and potential transcription factors. A multivariate analysis (canonical correspondence analyses) of various transcripts subdivided the incubated animals in three groups, one control group and two responding on short and long timescales, respectively. A strong dichotomy in the expression of peroxisome proliferator-activated receptors-1α and -ß receptors was most striking and has not previously been reported. Altogether, we identified a molecular network, which responds sensitively to warming beyond the realized ecological niche. The shift from lipid to carbohydrate stores and usage may support warm hardiness, as the latter sustain anaerobic metabolism and may prepare for hypoxemic conditions that would develop upon warming beyond the present acclimation temperature.


Assuntos
Aclimatação/genética , Regulação da Temperatura Corporal/genética , Temperatura Baixa , Metabolismo Energético/genética , Perfilação da Expressão Gênica , Temperatura Alta , Fígado/metabolismo , Perciformes/genética , Animais , Regiões Antárticas , Metabolismo dos Carboidratos/genética , Perfilação da Expressão Gênica/métodos , Regulação Enzimológica da Expressão Gênica , Metabolismo dos Lipídeos/genética , Fígado/enzimologia , Perciformes/metabolismo , Fatores de Tempo
11.
Harmful Algae ; 92: 101697, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32113604

RESUMO

Enrichment of the oceans with CO2 may be beneficial for some marine phytoplankton, including harmful algae. Numerous laboratory experiments provided valuable insights into the effects of elevated pCO2 on the growth and physiology of harmful algal species, including the production of phycotoxins. Experiments close to natural conditions are the next step to improve predictions, as they consider the complex interplay between biotic and abiotic factors that can confound the direct effects of ocean acidification. We therefore investigated the effect of ocean acidification on the occurrence and abundance of phycotoxins in bulk plankton samples during a long-term mesocosm experiment in the Gullmar Fjord, Sweden, an area frequently experiencing harmful algal blooms. During the experimental period, a total of seven phycotoxin-producing harmful algal genera were identified in the fjord, and in accordance, six toxin classes were detected. However, within the mesocosms, only domoic acid and the corresponding producer Pseudo-nitzschia spp. was observed. Despite high variation within treatments, significantly higher particulate domoic acid contents were measured in the mesocosms with elevated pCO2. Higher particulate domoic acid contents were additionally associated with macronutrient limitation. The risks associated with potentially higher phycotoxin levels in the future ocean warrants attention and should be considered in prospective monitoring strategies for coastal marine waters.


Assuntos
Fitoplâncton , Água do Mar , Concentração de Íons de Hidrogênio , Ácido Caínico/análogos & derivados , Estudos Prospectivos , Suécia
12.
BMC Bioinformatics ; 10: 41, 2009 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-19183437

RESUMO

BACKGROUND: Microsatellites (MSs) are DNA markers with high analytical power, which are widely used in population genetics, genetic mapping, and forensic studies. Currently available software solutions for high-throughput MS design (i) have shortcomings in detecting and distinguishing imperfect and perfect MSs, (ii) lack often necessary interactive design steps, and (iii) do not allow for the development of primers for multiplex amplifications. We present a set of new tools implemented as extensions to the STADEN package, which provides the backbone functionality for flexible sequence analysis workflows. The possibility to assemble overlapping reads into unique contigs (provided by the base functionality of the STADEN package) is important to avoid developing redundant markers, a feature missing from most other similar tools. RESULTS: Our extensions to the STADEN package provide the following functionality to facilitate microsatellite (and also minisatellite) marker design: The new modules (i) integrate the state-of-the-art tandem repeat detection and analysis software PHOBOS into workflows, (ii) provide two separate repeat detection steps - with different search criteria - one for masking repetitive regions during assembly of sequencing reads and the other for designing repeat-flanking primers for MS candidate loci, (iii) incorporate the widely used primer design program PRIMER3 into STADEN workflows, enabling the interactive design and visualization of flanking primers for microsatellites, and (iv) provide the functionality to find optimal locus- and primer pair combinations for multiplex primer design. Furthermore, our extensions include a module for storing analysis results in an SQLite database, providing a transparent solution for data access from within as well as from outside of the STADEN Package. CONCLUSION: The STADEN package is enhanced by our modules into a highly flexible, high-throughput, interactive tool for conventional and multiplex microsatellite marker design. It gives the user detailed control over the workflow, enabling flexible combinations of manual and automated analysis steps. The software is available under the OpenBSD License 12. The high efficiency of our automated marker design workflow has been confirmed in three microsatellite development projects.


Assuntos
Repetições de Microssatélites/genética , Repetições Minissatélites/genética , Análise de Sequência de DNA/métodos , Software , Animais , Primers do DNA , Bases de Dados Genéticas , Drosophila melanogaster/genética , Gastrópodes/genética , Marcadores Genéticos , Armazenamento e Recuperação da Informação , Internet , Camundongos , Interface Usuário-Computador
13.
J Comput Chem ; 30(3): 479-92, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18680215

RESUMO

A direct conformational clustering and mapping approach for peptide conformations based on backbone dihedral angles has been developed and applied to compare conformational sampling of Met-enkephalin using two molecular dynamics (MD) methods. Efficient clustering in dihedrals has been achieved by evaluating all combinations resulting from independent clustering of each dihedral angle distribution, thus resolving all conformational substates. In contrast, Cartesian clustering was unable to accurately distinguish between all substates. Projection of clusters on dihedral principal component (PCA) subspaces did not result in efficient separation of highly populated clusters. However, representation in a nonlinear metric by Sammon mapping was able to separate well the 48 highest populated clusters in just two dimensions. In addition, this approach also allowed us to visualize the transition frequencies between clusters efficiently. Significantly, higher transition frequencies between more distinct conformational substates were found for a recently developed biasing-potential replica exchange MD simulation method allowing faster sampling of possible substates compared to conventional MD simulations. Although the number of theoretically possible clusters grows exponentially with peptide length, in practice, the number of clusters is only limited by the sampling size (typically much smaller), and therefore the method is well suited also for large systems. The approach could be useful to rapidly and accurately evaluate conformational sampling during MD simulations, to compare different sampling strategies and eventually to detect kinetic bottlenecks in folding pathways.


Assuntos
Simulação por Computador , Encefalina Metionina/química , Modelos Químicos , Algoritmos , Conformação Proteica
14.
Sci Adv ; 5(4): eaav1110, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31032404

RESUMO

Dinoflagellates are microbial eukaryotes that have exceptionally large nuclear genomes; however, their organelle genomes are small and fragmented and contain fewer genes than those of other eukaryotes. The genus Amoebophrya (Syndiniales) comprises endoparasites with high genetic diversity that can infect other dinoflagellates, such as those forming harmful algal blooms (e.g., Alexandrium). We sequenced the genome (~100 Mb) of Amoebophrya ceratii to investigate the early evolution of genomic characters in dinoflagellates. The A. ceratii genome encodes almost all essential biosynthetic pathways for self-sustaining cellular metabolism, suggesting a limited dependency on its host. Although dinoflagellates are thought to have descended from a photosynthetic ancestor, A. ceratii appears to have completely lost its plastid and nearly all genes of plastid origin. Functional mitochondria persist in all life stages of A. ceratii, but we found no evidence for the presence of a mitochondrial genome. Instead, all mitochondrial proteins appear to be lost or encoded in the A. ceratii nucleus.


Assuntos
Dinoflagellida/genética , Dinoflagellida/metabolismo , Genoma Mitocondrial , Mitocôndrias/fisiologia , Filogenia , Aerobiose , Núcleo Celular/genética , Análise por Conglomerados , DNA Complementar/metabolismo , Evolução Molecular , Biblioteca Gênica , Genoma , Funções Verossimilhança , Microscopia Confocal , Análise de Sequência de DNA
15.
Appl Environ Microbiol ; 74(17): 5305-16, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18621863

RESUMO

The microalgal class Prasinophyceae (Chlorophyta) contains several picoeukaryotic species, which are known to be common in temperate and cold waters and have been observed to constitute major fractions of marine picoplankton. However, reliable detection and classification of prasinophytes are mainly hampered by their small size and few morphological markers. Consequently, very little is known about the abundance and ecology of the members of this class. In order to facilitate the assessment of the abundance of the Prasinophyceae, we have designed and evaluated an 18S rRNA gene-targeted oligonucleotide microarray consisting of 21 probes targeting different taxonomic levels of prasinophytes. The microarray contains both previously published probes from other hybridization methods and new probes, which were designed for novel prasinophyte groups. The evaluation of the probe set was done under stringent conditions with 18S PCR fragments from 20 unialgal reference cultures used as positive targets. This microarray has been applied to assess the community composition of prasinophytes at Helgoland, an island in the North Sea where time series data are collected and analyzed daily but only for the nano- and microplankton-size fractions. There is no identification of prasinophytes other than to record them numerically in the flagellate fraction. The samples were collected every 2 weeks between February 2004 and December 2006. The study here demonstrates the potential of DNA microarrays to be applied as a tool for quick general monitoring of this important picoplanktonic algal group.


Assuntos
Clorófitas/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Fitoplâncton/genética , Clorófitas/classificação , Sondas de DNA , DNA de Algas/genética , Mar do Norte , Filogenia , Fitoplâncton/classificação , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Sensibilidade e Especificidade
16.
Mol Ecol Resour ; 18(2): 204-216, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29024430

RESUMO

Next-generation sequencing is a common method for analysing microbial community diversity and composition. Configuring an appropriate sequence processing strategy within the variety of tools and methods is a nontrivial task and can considerably influence the resulting community characteristics. We analysed the V4 region of 18S rRNA gene sequences of marine samples by 454-pyrosequencing. Along this process, we generated several data sets with QIIME, mothur, and a custom-made pipeline based on DNAStar and the phylogenetic tree-based PhyloAssigner. For all processing strategies, default parameter settings and punctual variations were used. Our results revealed strong differences in total number of operational taxonomic units (OTUs), indicating that sequence preprocessing and clustering had a major impact on protist diversity estimates. However, diversity estimates of the abundant biosphere (abundance of ≥1%) were reproducible for all conducted processing pipeline versions. A qualitative comparison of diatom genera emphasized strong differences between the pipelines in which phylogenetic placement of sequences came closest to light microscopy-based diatom identification. We conclude that diversity studies using different sequence processing strategies are comparable if the focus is on higher taxonomic levels, and if abundance thresholds are used to filter out OTUs of the rare biosphere.


Assuntos
Biota , Biologia Computacional/métodos , Diatomáceas/classificação , Diatomáceas/genética , Análise de Sequência de DNA/métodos , Análise por Conglomerados , DNA Ribossômico/química , DNA Ribossômico/genética , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , RNA Ribossômico 18S/genética
18.
Sci Total Environ ; 605-606: 26-37, 2017 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-28662428

RESUMO

A diesel spill occurring at Carlini Station (King George Island (Isla 25 de Mayo), South Shetland Islands) in 2009 started the study of the fate of the hydrocarbons and their effect on the bacterial communities of the Potter Cove ecosystem. Soils and sediments were sampled across the 200-meter long diesel plume towards Potter Cove four and 15months after the spill. The sampling revealed a second fuel leakage from an underground pipeline at the spill site. The hydrocarbon fraction spilt over frozen and snow-covered ground reached the sea and dispersed with the currents. Contrary, diesel that infiltrated unfrozen soil remained detectable for years, and was seeping with ground water towards coastal marine sediments. Structural changes of the bacterial communities as well as hydrocarbon, carbon and nitrogen contents were investigated in sediments in front of the station, two affected terrestrial sites, and a terrestrial non-contaminated reference site. Bacterial communities (16S rRNA gene clone libraries) changed over time in contaminated soils and sediments. At the underground seepage site of highest contamination (5812 to 366µgg-1dw hydrocarbons from surface to 90-cm depth), communities were dominated by Actinobacteria (18%) and a betaproteobacterium closely related to Polaromonas naphthalenivorans (40%). At one of the spill sites, affected exclusively at the surface, contamination disappeared within one year. The same bacterial groups were enriched at both contaminated sites. This response at community level suggests that the cold-adapted indigenous microbiota in soils of the West Antarctic Peninsula have a high potential for bioremediation and can support soil cleaning actions in the ecosystem. Intensive monitoring of pollution and site assessment after episodic fuel spills is required for decision-making towards remediation strategies.


Assuntos
Bactérias/classificação , Sedimentos Geológicos/microbiologia , Poluição por Petróleo , Microbiologia do Solo , Solo/química , Regiões Antárticas , Bactérias/metabolismo , Biodegradação Ambiental , Hidrocarbonetos/química , Ilhas , RNA Ribossômico 16S/genética
19.
Sci Total Environ ; 574: 191-203, 2017 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-27636004

RESUMO

Cross-system studies on the response of different ecosystems to global change will support our understanding of ecological changes. Synoptic views on the planet's two main realms, the marine and terrestrial, however, are rare, owing to the development of rather disparate research communities. We combined questionnaires and a literature review to investigate how the importance of anthropogenic drivers of biodiversity change differs among marine and terrestrial systems and whether differences perceived by marine vs. terrestrial researchers are reflected by the scientific literature. This included asking marine and terrestrial researchers to rate the relevance of different drivers of global change for either marine or terrestrial biodiversity. Land use and the associated loss of natural habitats were rated as most important in the terrestrial realm, while the exploitation of the sea by fishing was rated as most important in the marine realm. The relevance of chemicals, climate change and the increasing atmospheric concentration of CO2 were rated differently for marine and terrestrial biodiversity respectively. Yet, our literature review provided less evidence for such differences leading to the conclusion that while the history of the use of land and sea differs, impacts of global change are likely to become increasingly similar.


Assuntos
Biodiversidade , Mudança Climática , Ecologia , Dióxido de Carbono/análise , Biologia Marinha
20.
ISME J ; 9(11): 2537-40, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25885562

RESUMO

Ice-binding proteins (IBPs) have been isolated from various sea-ice organisms. Their characterisation points to a crucial role in protecting the organisms in sub-zero environments. However, their in situ abundance and diversity in natural sea-ice microbial communities is largely unknown. In this study, we analysed the expression and phylogenetic diversity of eukaryotic IBP transcripts from microbial communities of Arctic and Antarctic sea ice. IBP transcripts were found in abundances similar to those of proteins involved in core cellular processes such as photosynthesis. Eighty-nine percent of the IBP transcripts grouped with known IBP sequences from diatoms, haptophytes and crustaceans, but the majority represented novel sequences not previously characterized in cultured organisms. The observed high eukaryotic IBP expression in natural eukaryotic sea ice communities underlines the essential role of IBPs for survival of many microorganisms in communities living under the extreme conditions of polar sea ice.


Assuntos
Camada de Gelo/microbiologia , Regiões Antárticas , Regiões Árticas , Diatomáceas/genética , Ecossistema , Geografia , Haptófitas , Camada de Gelo/química , Fotossíntese , Filogenia , Transcriptoma
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