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Title: Accurate, multi-kb reads resolve complex populations and detect rare microorganisms

Journal Article · · Genome Research
ORCiD logo [1];  [2];  [1];  [2];  [3];  [1];  [4];  [1];  [1];  [4];  [5];  [6]
  1. Univ. of California, Berkeley, CA (United States)
  2. Stanford Univ., CA (United States)
  3. Illumina Inc. Technology Development, Hayward, CA (United States)
  4. USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  6. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Accurate evaluation of microbial communities is essential for understanding global biogeochemical processes and can guide bioremediation and medical treatments. Metagenomics is most commonly used to analyze microbial diversity and metabolic potential, but assemblies of the short reads generated by current sequencing platforms may fail to recover heterogeneous strain populations and rare organisms. Here we used short (150-bp) and long (multi-kb) synthetic reads to evaluate strain heterogeneity and study microorganisms at low abundance in complex microbial communities from terrestrial sediments. The long-read data revealed multiple (probably dozens of) closely related species and strains from previously undescribed Deltaproteobacteria and Aminicenantes (candidate phylum OP8). Notably, these are the most abundant organisms in the communities, yet short-read assemblies achieved only partial genome coverage, mostly in the form of short scaffolds (N50 = ~2200 bp). Genome architecture and metabolic potential for these lineages were reconstructed using a new synteny-based method. Analysis of long-read data also revealed thousands of species whose abundances were <0.1% in all samples. Most of the organisms in this "long tail" of rare organisms belong to phyla that are also represented by abundant organisms. Genes encoding glycosyl hydrolases are significantly more abundant than expected in rare genomes, suggesting that rare species may augment the capability for carbon turnover and confer resilience to changing environmental conditions. Overall, the study showed that a diversity of closely related strains and rare organisms account for a major portion of the communities. These are probably common features of many microbial communities and can be effectively studied using a combination of long and short reads.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC02-05CH11231; SC0004918
OSTI ID:
1512117
Journal Information:
Genome Research, Vol. 25, Issue 4; ISSN 1088-9051
Publisher:
Cold Spring Harbor Laboratory PressCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 76 works
Citation information provided by
Web of Science

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Capturing One of the Human Gut Microbiome’s Most Wanted: Reconstructing the Genome of a Novel Butyrate-Producing, Clostridial Scavenger from Metagenomic Sequence Data journal May 2016
Lipid analysis of CO2-rich subsurface aquifers suggests an autotrophy-based deep biosphere with lysolipids enriched in CPR bacteria journal March 2020
cloudSPAdes: assembly of synthetic long reads using de Bruijn graphs journal July 2019
Identifying contamination with advanced visualization and analysis practices: metagenomic approaches for eukaryotic genome assemblies journal January 2016
The trajectory of microbial single-cell sequencing journal October 2017
Benchmarking viromics: an in silico evaluation of metagenome-enabled estimates of viral community composition and diversity journal January 2017
TruSPAdes: barcode assembly of TruSeq synthetic long reads journal February 2016
Beyond 16S rRNA Community Profiling: Intra-Species Diversity in the Gut Microbiota journal September 2016
Ecology of Subseafloor Crustal Biofilms journal August 2019
Utilization of defined microbial communities enables effective evaluation of meta-genomic assemblies journal April 2017
Long-read metagenomic exploration of extrachromosomal mobile genetic elements in the human gut journal August 2019
A viability-linked metagenomic analysis of cleanroom environments: eukarya, prokaryotes, and viruses journal December 2015
The past, present and future of microbiome analyses journal September 2016
Genome-Centric Analysis of Microbial Populations Enriched by Hydraulic Fracture Fluid Additives in a Coal Bed Methane Production Well journal June 2016
Genome-Centric Analysis of a Thermophilic and Cellulolytic Bacterial Consortium Derived from Composting journal April 2017
MetaSort untangles metagenome assembly by reducing microbial community complexity journal January 2017
Genome of the candidate phylum Aminicenantes bacterium from a deep subsurface thermal aquifer revealed its fermentative saccharolytic lifestyle journal January 2019
Genome assembly from synthetic long read clouds journal June 2016
Resolving the Complexity of Human Skin Metagenomes Using Single-Molecule Sequencing journal February 2016
Thousands of microbial genomes shed light on interconnected biogeochemical processes in an aquifer system journal October 2016
Unlinked rRNA genes are widespread among bacteria and archaea journal November 2019
Linking soil biology and chemistry in biological soil crust using isolate exometabolomics journal January 2018
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Microbial community composition along a 50 000-year lacustrine sediment sequence journal February 2018
Mediterranean grassland soil C–N compound turnover is dependent on rainfall and depth, and is mediated by genomically divergent microorganisms journal May 2019
SPAligner: Alignment of Long Diverged Molecular Sequences to Assembly Graphs posted_content August 2019
Critical biogeochemical functions in the subsurface are associated with bacteria from new phyla and little studied lineages: N- and C-cycling organisms in the subsurface journal July 2015
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Interpreting Microbial Biosynthesis in the Genomic Age: Biological and Practical Considerations journal June 2017
Aquifer environment selects for microbial species cohorts in sediment and groundwater journal February 2015
Emerging spatial patterns in Antarctic prokaryotes journal September 2015
Long-read based de novo assembly of low-complexity metagenome samples results in finished genomes and reveals insights into strain diversity and an active phage system journal June 2019
metaSPAdes: a new versatile metagenomic assembler journal March 2017
Functional Metagenomics as a Tool for Identification of New Antibiotic Resistance Genes from Natural Environments journal October 2016
Mining prokaryotes for antimicrobial compounds: from diversity to function journal April 2017
Structure and function of the global topsoil microbiome journal August 2018
A Meta-proteogenomic Approach to Peptide Identification Incorporating Assembly Uncertainty and Genomic Variation journal May 2019
Proteogenomic analyses indicate bacterial methylotrophy and archaeal heterotrophy are prevalent below the grass root zone journal January 2016
Anvi’o: an advanced analysis and visualization platform for ‘omics data journal January 2015
Synthetic long-read sequencing reveals intraspecies diversity in the human microbiome journal January 2016
Identification of Major Rhizobacterial Taxa Affected by a Glyphosate-Tolerant Soybean Line via Shotgun Metagenomic Approach journal April 2018
Shotgun metagenomics, from sampling to analysis journal September 2017
Linking Associations of Rare Low-Abundance Species to Their Environments by Association Networks journal March 2018
Metatranscriptomic reconstruction reveals RNA viruses with the potential to shape carbon cycling in soil journal November 2019
Haplotype-Phased Synthetic Long Reads from Short-Read Sequencing journal January 2016
Tracking Strains in the Microbiome: Insights from Metagenomics and Models journal May 2016