Methyl-compound use and slow growth characterize microbial life in 2-km-deep subseafloor coal and shale beds

Proceedings of the National Academy of Sciences of the United States of America
E Trembath-ReichertVictoria J Orphan

Abstract

The past decade of scientific ocean drilling has revealed seemingly ubiquitous, slow-growing microbial life within a range of deep biosphere habitats. Integrated Ocean Drilling Program Expedition 337 expanded these studies by successfully coring Miocene-aged coal beds 2 km below the seafloor hypothesized to be "hot spots" for microbial life. To characterize the activity of coal-associated microorganisms from this site, a series of stable isotope probing (SIP) experiments were conducted using intact pieces of coal and overlying shale incubated at in situ temperatures (45 °C). The 30-month SIP incubations were amended with deuterated water as a passive tracer for growth and different combinations of 13C- or 15N-labeled methanol, methylamine, and ammonium added at low (micromolar) concentrations to investigate methylotrophy in the deep subseafloor biosphere. Although the cell densities were low (50-2,000 cells per cubic centimeter), bulk geochemical measurements and single-cell-targeted nanometer-scale secondary ion mass spectrometry demonstrated active metabolism of methylated substrates by the thermally adapted microbial assemblage, with differing substrate utilization profiles between coal and shale incubations. The conversion ...Continue Reading

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Methods Mentioned

BETA
overprinting
fluorescence-activated cell sorting
FACS
chips
flow cytometry
dissection
PCR
flow
electrophoresis

Software Mentioned

R
Look
Qiime
gridExtra
@ NanoSIMS
ggplot2
NanoSIMS
dplyr
RColorBrewer

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