Abstract:
:Growing appreciation for the biogeochemical significance of uncultured microorganisms is changing the focus of environmental microbiology. Techniques designed to investigate microbial metabolism in situ are increasingly popular, from mRNA-targeted fluorescence in situ hybridization (FISH) to the "-omics" revolution, including metagenomics, transcriptomics, and proteomics. Recently, the coupling of FISH with nanometer-scale secondary ion mass spectrometry (NanoSIMS) has taken this movement in a new direction, allowing single-cell metabolic analysis of uncultured microbial phylogenic groups. The main advantage of FISH-NanoSIMS over previous noncultivation-based techniques to probe metabolism is its ability to directly link 16S rRNA phylogenetic identity to metabolic function. In the following chapter, we describe the procedures necessary to identify nitrogen-fixing microbes within marine sediment via FISH-NanoSIMS, using our work on nitrogen fixation by uncultured deep-sea methane-consuming archaea as a case study.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Dekas AE,Orphan VJdoi
10.1016/B978-0-12-381294-0.00012-2subject
Has Abstractpub_date
2011-01-01 00:00:00pages
281-305eissn
0076-6879issn
1557-7988pii
B978-0-12-381294-0.00012-2journal_volume
486pub_type
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