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915 results for “metagenomics”
Data for "Analysis of metagenome-assembled viral genomes from the human gut reveals diverse putative CrAss-like phages with unique genomic features"
<p>Data for "Analysis of metagenome-assembled viral genomes from the human gut reveals diverse putative CrAss-like phages with unique genomic features" (submitted to Nature Communications)</p>
Estimation of the relative abundance of species in artificial mixtures of insects using low-coverage shotgun metagenomics
<p>Amplicon metabarcoding is an established technique to analyse the taxonomic composition of communities of organisms using high-throughput DNA sequencing, but there are doubts about its ability to quantify the relative proportions of the species, as opposed to the species list. Here, we bypass the enrichment step and avoid the PCR-bias, by directly sequencing the extracted DNA using shotgun metagenomics. This approach is common practice in prokaryotes, but not in eukaryotes, because of the low number of sequenced genomes of eukaryotic species. We tested the metagenomics approach using insect species whose genome is already sequenced and assembled to an advanced degree. We shotgun-sequenced, at low-coverage DNA, 18 species of insects in 22 single-species and 6 mixed-species libraries and mapped the reads against 110 reference genomes of insects. We used the single-species libraries to calibrate the process of assignation of reads to species and the libraries created from species mixtures to evaluate the ability of the method to quantify the relative species abundance. Our results showed that the shotgun metagenomic method is easily able to set apart closely-related insect species, like four species of <i>Drosophila</i> included in the artificial libraries. However, to avoid the counting of rare misclassified reads in samples, it was necessary to use a rather stringent detection limit of 0.001, so species with a lower relative abundance are ignored. We also identified that approximately half the raw reads were informative for taxonomic purposes. Finally, using the mixed-species libraries, we showed that it was feasible to quantify with confidence the relative abundance of individual species in the mixtures.</p>
Data from: Mining for NRPS and PKS genes revealed a high diversity in the Sphagnum bog metagenome
Sphagnum bog ecosystems are among the oldest vegetation forms harboring a specific microbial community and are known to produce an exceptionally wide variety of bioactive substances. Although the Sphagnum metagenome shows a rich secondary metabolism, the genes have not yet been explored. To analyze nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs), the diversity of NRPS and PKS genes in Sphagnum-associated metagenomes was investigated by in silico data mining and sequence-based screening (PCR amplification of 9,500 fosmid clones). The in silico Illumina-based metagenomic approach resulted in the identification of 279 NRPSs and 346 PKSs, as well as 40 PKS-NRPS hybrid gene sequences. The occurrence of NRPS sequences was strongly dominated by the members of the Protebacteria phylum, especially by species of the Burkholderia genus, while PKS sequences were mainly affiliated with Actinobacteria. Thirteen novel NRPS-related sequences were identified by PCR amplification screening, displaying amino acid identities of 48% to 91% to annotated sequences of members of the phyla Proteobacteria, Actinobacteria, and Cyanobacteria. Some of the identified metagenomic clones showed the closest similarity to peptide synthases from Burkholderia or Lysobacter, which are emerging bacterial sources of as-yet-undescribed bioactive metabolites. This report highlights the role of the extreme natural ecosystems as a promising source for detection of secondary compounds and enzymes, serving as a source for biotechnological applications.
Data from: Stalking the fourth domain in metagenomic data: searching for, discovering, and interpreting novel, deep branches in phylogenetic trees of phylogenetic marker genes
BACKGROUND: Most of our knowledge about the ancient evolutionary history of organisms has been derived from data associated with specific known organisms (i.e., organisms that we can study directly such as plants, metazoans, and culturable microbes). Recently, however, a new source of data for such studies has arrived: DNA sequence data generated directly from environmental samples. Such metagenomic data has enormous potential in a variety of areas including, as we argue here, in studies of very early events in the evolution of gene families and of species. METHODOLOGY/PRINCIPAL FINDINGS: We designed and implemented new methods for analyzing metagenomic data and used them to search the Global Ocean Sampling (GOS) Expedition data set for novel lineages in three gene families commonly used in phylogenetic studies of known and unknown organisms: small subunit rRNA and the recA and rpoB superfamilies. Though the methods available could not accurately identify very deeply branched ss-rRNAs (largely due to difficulties in making robust sequence alignments for novel rRNA fragments), our analysis revealed the existence of multiple novel branches in the recA and rpoB gene families. Analysis of available sequence data likely from the same genomes as these novel recA and rpoB homologs was then used to further characterize the possible organismal source of the novel sequences. CONCLUSIONS/SIGNIFICANCE: Of the novel recA and rpoB homologs identified in the metagenomic data, some likely come from uncharacterized viruses while others may represent ancient paralogs not yet seen in any cultured organism. A third possibility is that some come from novel cellular lineages that are only distantly related to any organisms for which sequence data is currently available. If there exist any major, but so-far-undiscovered, deeply branching lineages in the tree of life, we suggest that methods such as those described herein currently offer the best way to search for them.
Data from: Using metagenomics to show the efficacy of forest restoration in the New Jersey Pine Barrens
The Franklin Parker Preserve within the New Jersey Pine Barrens contains 5,000 acres of wetlands habitat, including old-growth Atlantic White Cedar (or AWC; Chamaecyparis thyoides) swamps, cranberry bogs, and former cranberry bogs undergoing restoration into AWC forests. This study showed that the C-use efficiency was greater in the old-growth AWC soils than in soils from 8-year old mid-stage restored AWC stands, which were greater than found in soil from 4-year old AWC stands—the latter two stands being restored from long-term cranberry bogs. A metagenomic analysis of eDNA extracted from these soils showed that the C-cycle trends were associated with increases in the relative numbers of DNA sequences from several copiotrophic bacterial groups (Bacteroidetes, and Proteobacteria), complex C decomposing fungal groups ( Sordiomycetes, Mortierellales, and Thelephorales), and collembolan and formicid invertebrates. All groups are indicators of successionally more advanced soils, and critical for soil C-cycle activities. These data suggest that the restoration activities studied are enhancing critical guilds of soil biota, and increasing C-use efficiency in the soils of restored habitats, and that the use of metagenomic analysis of soil eDNA can be used in the development of assessment models for soil recovery of wetlands following restoration.
Data from: Metabarcoding and mitochondrial metagenomics of endogean arthropods to unveil the mesofauna of the soil
Biological communities inhabiting the soil are among the most diversified, complex and yet most poorly studied terrestrial ecosystems. The greatest knowledge gaps apply to the arthropod mesofauna (0·1–2 mm body size) because conventional morphological and molecular approaches are in many cases insufficient for the characterisation of these complex communities. The development of high-throughput sequencing (HTS) methodologies is required to solve current impediments and to further advance our understanding of below-ground biodiversity. We propose a flotation–Berlese–flotation (FBF) protocol for sampling and specimen processing to obtain 'clean' DNA extractions of arthropod mesofauna from the soil. In addition, we developed and tested HTS protocols for the characterisation of arthropod communities from these bulk DNA extractions using cox1 metabarcoding and shotgun metagenomic sequencing on the MiSeq Illumina platform. The FBF protocol provided DNA of soil arthropods from sufficiently large volumes of soil and free from contaminating bacteria and inhibitors. Metabarcoding and metagenomic sequencing on two deep soil samples from Iberian grasslands revealed >100 species of Acari and Collembola from 28 families. Genome assembly straight from shotgun sequencing of bulk specimens produced partial and full mitogenomes for 54 species with average length of >6000 bp. Metabarcoding and metagenomic sequencing resulted in closely congruent OTUs, but species numbers were highest with metabarcoding, while ∼73% of species were confirmed by matching shotgun sequence reads and ∼48% by contig assembly from those shotgun reads. In combination, the FBF protocol together with the PCR-based and shotgun sequencing pipelines addressed most of the challenges of studying soil arthropod mesofauna on the MiSeq Illumina platform. They are powerful, cost-efficient tools for characterising soil diversity in a phylogenetic and community ecology context. These methodological developments of HTS approaches for the study of mesofauna will accelerate ecological and evolutionary studies, biomonitoring of soil arthropods, and progress in both theoretical and applied soil science.
Data from: Metagenomic chromosome conformation capture (meta3C) unveils the diversity of chromosome organization in microorganisms
Genomic analyses of microbial populations in their natural environment remain limited by the difficulty to assemble full genomes of individual species. Consequently, the chromosome organization of microorganisms has been investigated in a few model species, but the extent to which the features described can be generalized to other taxa remains unknown. Using controlled mixes of bacterial and yeast species, we developed meta3C, a metagenomic chromosome conformation capture approach that allows characterizing individual genomes and their average organization within a mix of organisms. Not only can meta3C be applied to species already sequenced, but a single meta3C library can be used for assembling, scaffolding and characterizing the tridimensional organization of unknown genomes. By applying meta3C to a semi-complex environmental sample, we confirmed its promising potential. Overall, this first meta3C study highlights the remarkable diversity of microorganisms chromosome organization, while providing an elegant and integrated approach to metagenomic analysis. - See more at: http://elifesciences.org/content/3/e03318#sthash.Yx6nSY4J.dpuf
Data from: Validating the power of mitochondrial metagenomics for community ecology and phylogenetics of complex assemblages
1. The biodiversity of mixed-species samples of arthropods can be characterized by shotgun sequencing of bulk genomic DNA and subsequent bioinformatics assembly of mitochondrial genomes. Here, we tested the power of mitochondrial metagenomics by conducting Illumina sequencing on mixtures of >2600 individuals of leaf beetles (Chrysomelidae) from 10 communities. 2. Patterns of species richness, community dissimilarity and biomass were assessed from matches of reads against three reference databases, including (i) a custom set of mitogenomes generated for 156 species (89% of species in the study); (ii) mitogenomes obtained by the de novo assembly of sequence reads from the real-world communities; and (iii) a custom set of DNA barcode (cox1-5′) sequences. 3. Species detection against the custom-built reference genomes was very high (>90%). False presences were rare against mitogenomes but slightly higher against the barcode references. False absences were mainly due to the incompleteness of the reference databases and, thus, more prevalent in the de novo data set. Biomass (abundance × body length) and read numbers were strongly correlated, demonstrating the potential of mitochondrial metagenomics for studies of species abundance. 4. A phylogenetic tree from the mitogenomes showed high congruence with known relationships in Chrysomelidae. Patterns of taxonomic and phylogenetic dissimilarity between sites were highly consistent with data from morphological identifications. 5. The power of mitochondrial metagenomics results from the possibility of rapid assembly of mitogenomes from mixtures of specimens and the use of read counts for accurate estimates of key parameters of biodiversity directly from community samples.
Metagenomic data for Bathymodiolus symbionts deposited in IMG
<p>Metagenomic data for the sulfur- and methane-oxidizing symbionts of <em>Bathymodiolus</em> mussels deposited in the Integrated Microbial Genomes (IMG) database of the DOE Joint Genome Institute (http://img.jgi.doe.gov/)</p>
Metagenomic data for Bathymodiolus symbionts deposited in IMG (2016)
<p>Metagenomic data for the sulfur- and methane-oxidizing symbionts of <em>Bathymodiolus</em> mussels deposited in the Integrated Microbial Genomes (IMG) database of the DOE Joint Genome Institute (http://img.jgi.doe.gov/) until October 2016.</p>
Benchmarking datasets used in the manuscript "Strain-level metagenomic profiling using pangenome graphs with PanTax"
Open the record for dataset details and reuse information.
Updated Metagenomic Species Pan-genomes (MSPs) of the human gastrointestinal microbiota
<p></p><h1>Gene catalog construction</h1><br>The methodology for creating the IGC2 catalog is described in the original papers: Li et al., 2014 and Wen et al., 2017<br><h1>MSP creation</h1><br>Reads from publicly available human gut metagenomes were aligned against the IGC2 catalog with the Meteor to produce a raw gene abundance table (10.4M genes quantified in >2000 samples). Then, co-abundant genes were binned in 1,989 Metagenomic Species Pan-genomes (MSPs, i.e. clusters of co-abundant genes that likely belong to the same microbial species) using MSPminer.<br><h1>MSPs taxonomic annotation</h1><br>MSPs taxonomic annotation was performed by aligning MSP core and accessory genes against representative genomes of the Genome Taxonomy Database (GTDB r207) using blastn (task = megablast, word_size = 16). The 20 best hits for each gene were kept (--max-target-seq 20). Using an in-house pipeline, a species-level assignment was given if > 50% of the genes matched the representative genome of a given species, with a mean identity ≥ 95% and mean gene length coverage ≥ 90%. The remaining MSPs were assigned to a higher taxonomic level (genus to superkingdom), if more than 50% of their genes had the same annotation.<br><h1>Construction of the phylogenetic tree</h1><br>39 universal phylogenetic markers genes were extracted from the MSPs with fetchMGs. Then, the markers were separately aligned with MUSCLE. The alignments were merged and trimmed with trimAl (parameters: -automated1). Finally, the phylogenetic tree was computed with FastTreeMP (parameters: -gamma -pseudo -spr -mlacc 3 -slownni). <h1>Mapping rate distribution across public cohorts</h1>We generated mapping rate distribution plots using Meteor2 (default parameters), comparing performance between: PRJEB1786, PRJEB5224, PRJEB6337, PRJNA422434 (cohort used in catalogue assembly) and PRJEB11532, PRJEB33500, PRJEB37249, PRNJNA834801 (independent cohort not used in assembly).<p></p>
MetaChick: characterization of the chicken caecal metagenome by deep shotgun sequencing
<p></p><h1>Data sources</h1><br>This dataset was constructed using the samples of the MetaChick project (phase 1) corresponding to the cecal content of 340 animals. Sequencing data and associated metadata have been submitted to INSDC (bioproject: PRJEB38174).<br><h1>Sequencing data QC and metagenomic assembly</h1><br>First, sequencing adapters removal and read trimming was performed with fastxtend. Reads mapped on the host genome (GRCg7b GCA_016699485.1) with bowtie2 were removed with samtools. Finally, metagenomic assembly was performed with metaSPAdes v3.14.1. Contigs of less than 1500 bp were removed.<br><h1>MAGs recovery</h1><br>MAGs were generated with MetaBAT 2 (multi-coverage mode) and MAGs quality was assessed with CheckM. MAGs with completeness < 70% or contamination > 5% or N50 < 8Kb were discarded. Pairwise Average Nucleotide Identity (ANI) was computed for all recovered MAGs with fastANI and dereplication at species level (ANI cutoff = 95%).<br><h1>Non-redundant gene catalog</h1><br>Genes were predicted on all contigs from metagenomic assemblies with Prodigal (parameters : -m -p meta). Genes were pooled and clustered with cd-hit-est (parameters -c 0.95 -aS 0.90 -G 0 -d 0 -M 0 -T 0) by choosing those from the longest contigs as representatives.<br><h1>MSPs recovery</h1><br>A raw gene abundance table (13,6M genes quantified in 340 samples) was generated with meteorMeteor. Then, co-abundant genes were binned in Metagenomic Species Pan-genomes (MSPs, i.e. gene clusters that likely belong to the same microbial species) using MSPminer.<br><h1>MAGs and MSPs taxonomic annotation</h1><br>Dereplicated MAGs were annotated with GTDB-Tk based on GTDB r214. Then, MAGs taxonomic annotation was propagated to the corresponding MSPs.<br><h1>Construction of the phylogenetic tree</h1><br>39 universal phylogenetic markers genes were extracted from the dereplicated MAGs with fetchMGs. Then, the markers were separately aligned with MUSCLE. The 40 alignments were merged and trimmed with trimAl (parameters: -automated1). Finally, the phylogenetic tree was computed with FastTreeMP (parameters: -gamma -pseudo -spr -mlacc 3 -slownni).<h1>Mapping rate distribution across public cohorts</h1>We generated mapping rate distribution plots using Meteor2 (default parameters), comparing performance between: PRJEB38174 (cohort used in catalogue assembly) and PRJEB29033, PRJEB33338, PRJEB53667 (independent cohort not used in assembly).<p></p>
MicroReset: characterization of the rabbit (Oryctolagus cuniculus) fecal metagenome and resistome by deep shotgun sequencing
<p></p><h1>Data source</h1><br>The dataset was generated from 30 rabbit fecal samples subjected to deep shotgun metagenomic sequencing. The sequencing data is available under the BioProject PRJEB50625.<br>Metagenomic Assembly<br>Raw sequencing reads were first pre-processed using fastp for adapter removal and quality trimming. Host-derived reads were filtered out by mapping to the rabbit reference genome (GCF_000001635.27) using Bowtie2 and removing mapped reads with Samtools. Each sample was individually assembled using metaSPAdes. Contigs shorter than 1,500 bp were excluded from downstream analysis.<br><h1>MAG Recovery</h1><br>Reads from each sample were mapped to all 30 assemblies (30×30 mappings) using Bowtie2. The resulting alignments were sorted and indexed with Samtools. Contig coverage across all samples was computed using `jgi_summarize_bam_contig_depths`. Binning was performed with MetaBAT 2 and SemiBin v1.3. MAG quality was assessed with CheckM. Only high-quality MAGs (≥70% completeness, ≤5% contamination, N50 ≥ 8 kb) were retained.<br>Non-Redundant Gene Catalog<br>Gene prediction was carried out using Prodigal on all contigs from the current study (with `-m -p meta`). Genes shorter than 90 bp or lacking start/stop codons were discarded. The remaining genes from both sources were pooled and clustered using CD-HIT-EST (parameters: `-c 0.95 -aS 0.90 -G 0 -d 0 -M 0 -T 0`). The longest contigs were used to select representative genes.<br><h1>MSP Recovery</h1><br>Shotgun reads from the 30 samples were aligned to the non-redundant gene catalog using the Meteor suite, generating a gene abundance matrix (5.7 million genes × 30 samples). Co-abundant genes were grouped into 1,053 Metagenomic Species Pan-genomes (MSPs) using MSPminer.<br><h1>Taxonomic Annotation of MSPs</h1><br>MAGs representing each species were taxonomically annotated using GTDB-Tk with GTDB release r214. The resulting taxonomy was propagated to the corresponding MSPs.<br><h1>Phylogenetic Tree Construction</h1><br>A set of 39 universal phylogenetic marker genes was extracted from the 1,053 MSPs (or their corresponding MAGs, when available) using fetchMGs. Each marker was independently aligned using MUSCLE, and the alignments were concatenated and trimmed using trimAl (parameter: `-automated1`). A maximum-likelihood phylogenetic tree was constructed with FastTreeMP (parameters: `-gamma -pseudo -spr -mlacc 3 -slownni`).<h1>Mapping rate distribution across public cohorts</h1>We generated mapping rate distribution plots using Meteor2 (default parameters) for PRJEB50625 (cohort used in catalogue assembly).<p></p>
Required data to regenerate simulated metagenomes and reproduce KO-identification results in these simulated metagenomes
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Supplementary material 12 from: Paez-Triana L, Herrera G, Vega L, Garcia-Corredor D, Pulido Medellín MO, Paniz-Mondolfi A, Muñoz M, Ramírez JD (2023) Metagenomic exploration of endosymbionts and pathogens in the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.) ticks in Colombia. Metabarcoding and Metagenomics 7: e109085. https://doi.org/10.3897/mbmg.7.109085
p-values obtained between departments and sexes within each department using the Wilcoxon test on the relative abundances of pathogens and endosymbionts
Supplementary material 3 from: Paez-Triana L, Herrera G, Vega L, Garcia-Corredor D, Pulido Medellín MO, Paniz-Mondolfi A, Muñoz M, Ramírez JD (2023) Metagenomic exploration of endosymbionts and pathogens in the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.) ticks in Colombia. Metabarcoding and Metagenomics 7: e109085. https://doi.org/10.3897/mbmg.7.109085
Significant differences in the relative abundances of assigned reads in Bacteria and Archaea between departments and sexes of the various genera found in the samples
Supplementary material 7 from: Paez-Triana L, Herrera G, Vega L, Garcia-Corredor D, Pulido Medellín MO, Paniz-Mondolfi A, Muñoz M, Ramírez JD (2023) Metagenomic exploration of endosymbionts and pathogens in the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.) ticks in Colombia. Metabarcoding and Metagenomics 7: e109085. https://doi.org/10.3897/mbmg.7.109085
Metadata and information regarding the genomes included in the analysis for endosymbiont and pathogen identification
Supplementary material 5 from: Paez-Triana L, Herrera G, Vega L, Garcia-Corredor D, Pulido Medellín MO, Paniz-Mondolfi A, Muñoz M, Ramírez JD (2023) Metagenomic exploration of endosymbionts and pathogens in the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.) ticks in Colombia. Metabarcoding and Metagenomics 7: e109085. https://doi.org/10.3897/mbmg.7.109085
Results of dereplication, as well as phylogenomics analysis and pangenome exploration of all samples (including longer MAGs)
Supplementary material 10 from: Paez-Triana L, Herrera G, Vega L, Garcia-Corredor D, Pulido Medellín MO, Paniz-Mondolfi A, Muñoz M, Ramírez JD (2023) Metagenomic exploration of endosymbionts and pathogens in the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.) ticks in Colombia. Metabarcoding and Metagenomics 7: e109085. https://doi.org/10.3897/mbmg.7.109085
p-values obtained between departments and sexes within each department using the Wilcoxon test on the relative abundances of each genus (Bacteria and Archaea)
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