Abstract
Reference genomes are required to understand the diverse roles of microorganisms in ecology, evolution, human and animal health, but most species remain uncultured. Here we present a sequence composition–independent approach to recover high-quality microbial genomes from deeply sequenced metagenomes. Multiple metagenomes of the same community, which differ in relative population abundances, were used to assemble 31 bacterial genomes, including rare (<1% relative abundance) species, from an activated sludge bioreactor. Twelve genomes were assembled into complete or near-complete chromosomes. Four belong to the candidate bacterial phylum TM7 and represent the most complete genomes for this phylum to date (relative abundances, 0.06–1.58%). Reanalysis of published metagenomes reveals that differential coverage binning facilitates recovery of more complete and higher fidelity genome bins than other currently used methods, which are primarily based on sequence composition. This approach will be an important addition to the standard metagenome toolbox and greatly improve access to genomes of uncultured microorganisms.
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NCBI Reference Sequence
Sequence Read Archive
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Acknowledgements
This study was funded by Aalborg University and the Danish Research Council for Strategic Research via the Centre “EcoDesign-MBR” and the Obelske Family foundation. P.H. was supported by a Discovery Outstanding Researcher Award (DORA) from the Australian Research Council, grant DP120103498 and G.W.T. was supported by a QEII fellowship from the Australian Research Council, grant DP1093175. We thank S. McIlroy and P. Larsen for assistance with FISH analyses, A.M. Saunders for 16S rRNA data generation and J.P. Euzeby for suggesting the new genus name.
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M.A., experimental design, data analysis and manuscript; P.H., data analysis and manuscript; A.S., data analysis; K.L.N., sequencing; G.W.T., data analysis and manuscript; P.H.N., experimental design and manuscript.
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Supplementary Notes, Supplementary Figures 1–13 and Supplementary Tables 1–10 (PDF 4591 kb)
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All scripts used in the manuscript, including a detailed step by step guide and example datasets. (ZIP 7822 kb)
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Albertsen, M., Hugenholtz, P., Skarshewski, A. et al. Genome sequences of rare, uncultured bacteria obtained by differential coverage binning of multiple metagenomes. Nat Biotechnol 31, 533–538 (2013). https://doi.org/10.1038/nbt.2579
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DOI: https://doi.org/10.1038/nbt.2579
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