Dataset Information

Dissecting biological "dark matter" with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth.

ABSTRACT: We have developed a microfluidic device that allows the isolation and genome amplification of individual microbial cells, thereby enabling organism-level genomic analysis of complex microbial ecosystems without the need for culture. This device was used to perform a directed survey of the human subgingival crevice and to isolate bacteria having rod-like morphology. Several isolated microbes had a 16S rRNA sequence that placed them in candidate phylum TM7, which has no cultivated or sequenced members. Genome amplification from individual TM7 cells allowed us to sequence and assemble >1,000 genes, providing insight into the physiology of members of this phylum. This approach enables single-cell genetic analysis of any uncultivated minority member of a microbial community.

SUBMITTER: Marcy Y

PROVIDER: S-EPMC1924555 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature

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Dissecting biological "dark matter" with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth.

Marcy Yann Y Ouverney Cleber C Bik Elisabeth M EM Lösekann Tina T Ivanova Natalia N Martin Hector Garcia HG Szeto Ernest E Platt Darren D Hugenholtz Philip P Relman David A DA Quake Stephen R SR

Proceedings of the National Academy of Sciences of the United States of America 20070709 29

We have developed a microfluidic device that allows the isolation and genome amplification of individual microbial cells, thereby enabling organism-level genomic analysis of complex microbial ecosystems without the need for culture. This device was used to perform a directed survey of the human subgingival crevice and to isolate bacteria having rod-like morphology. Several isolated microbes had a 16S rRNA sequence that placed them in candidate phylum TM7, which has no cultivated or sequenced mem ...[more]

PMID: 17620602

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Project description:In the late 20th century, cosmology became a precision science. Now, at the beginning of the next century, the parameters describing how our universe evolved from the Big Bang are generally known to a few percent. One key parameter is the total mass density of the universe. Normal matter constitutes only a small fraction of the total mass density. Observations suggest this additional mass, the dark matter, is cold (that is, moving nonrelativistically in the early universe) and interacts feebly if at all with normal matter and radiation. There's no known such elementary particle, so the strong presumption is the dark matter consists of particle relics of a new kind left over from the Big Bang. One of the most important questions in science is the nature of this dark matter. One attractive particle dark-matter candidate is the axion. The axion is a hypothetical elementary particle arising in a simple and elegant extension to the standard model of particle physics that nulls otherwise observable CP-violating effects (where CP is the product of charge reversal C and parity inversion P) in quantum chromo dynamics (QCD). A light axion of mass 10(-(6-3)) eV (the invisible axion) would couple extraordinarily weakly to normal matter and radiation and would therefore be extremely difficult to detect in the laboratory. However, such an axion is a compelling dark-matter candidate and is therefore a target of a number of searches. Compared with other particle dark-matter candidates, the plausible range of axion dark-matter couplings and masses is narrowly constrained. This focused search range allows for definitive searches, where a nonobservation would seriously impugn the dark-matter QCD-axion hypothesis. Axion searches use a wide range of technologies, and the experiment sensitivities are now reaching likely dark-matter axion couplings and masses. This article is a selective overview of the current generation of sensitive axion searches. Not all techniques and experiments are discussed, but I hope to give a sense of the current experimental landscape of the search for dark-matter axions.

Project description:Particles in aquatic environments host distinct communities of microbes, yet the evolution of particle-specialized taxa and the extent to which specialized microbial metabolism is associated with particles is largely unexplored. Here, we investigate the hypothesis that a widely distributed and uncultivated microbial group--the marine group II euryarchaea (MGII)--interacts with living and detrital particulate organic matter (POM) in the euphotic zone of the central California Current System. Using fluorescent in situ hybridization, we verified the association of euryarchaea with POM. We further quantified the abundance and distribution of MGII 16?S ribosomal RNA genes in size-fractionated seawater samples and compared MGII functional capacity in metagenomes from the same fractions. The abundance of MGII in free-living and >3??m fractions decreased with increasing distance from the coast, whereas MGII abundance in the 0.8-3??m fraction remained constant. At several offshore sites, MGII abundance was highest in particle fractions, indicating that particle-attached MGII can outnumber free-living MGII under oligotrophic conditions. Compared with free-living MGII, the genome content of MGII in particle-associated fractions exhibits an increased capacity for surface adhesion, transcriptional regulation and catabolism of high molecular weight substrates. Moreover, MGII populations in POM fractions are phylogenetically distinct from and more diverse than free-living MGII. Eukaryotic phytoplankton additions stimulated MGII growth in bottle incubations, providing the first MGII net growth rate measurements. These ranged from 0.47 to 0.54?d(-1). However, MGII were not recovered in whole-genome amplifications of flow-sorted picoeukaryotic phytoplankton and heterotrophic nanoflagellates, suggesting that MGII in particle fractions are not physically attached to living POM. Collectively, our results support a linkage between MGII ecophysiology and POM, implying that marine archaea have a role in elemental cycling through interactions with particles.

Dataset Information

Dissecting biological "dark matter" with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth.

Publications

Dissecting biological "dark matter" with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth.

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