Tracing sources of atmospheric methane using clumped isotopes.

We apply a recently developed measurement technique for methane (CH4) isotopologues* (isotopic variants of CH4-13CH4, 12CH3D, 13CH3D, and 12CH2D2) to identify contributions to the atmospheric burden from fossil fuel and microbial sources. The aim of this study is to constrain factors that ultimately control the concentration of this potent greenhouse gas on global, regional, and local levels. While predictions of atmospheric methane isotopologues have been modeled, we present direct measurements that point to a different atmospheric methane composition and to a microbial flux with less clumping (greater deficits relative to stochastic) in both 13CH3D and 12CH2D2 than had been previously assigned. These differences make atmospheric isotopologue data sufficiently sensitive to variations in microbial to fossil fuel fluxes to distinguish between emissions scenarios such as those generated by different versions of EDGAR (the Emissions Database for Global Atmospheric Research), even when existing constraints on the atmospheric CH4 concentration profile as well as traditional isotopes are kept constant.

Well-hidden methanogenesis in deep, organic-rich sediments of Guaymas Basin.

Deep marine sediments (>1mbsf) harbor ~26% of microbial biomass and are the largest reservoir of methane on Earth. Yet, the deep subsurface biosphere and controls on its contribution to methane production remain underexplored. Here, we use a multidisciplinary approach to examine methanogenesis in sediments (down to 295 mbsf) from sites with varying degrees of thermal alteration (none, past, current) at Guaymas Basin (Gulf of California) for the first time. Traditional (13C/12C and D/H) and multiply substituted (13CH3D and 12CH2D2) methane isotope measurements reveal significant proportions of microbial methane at all sites, with the largest signal at the site with past alteration. With depth, relative microbial methane decreases at differing rates between sites. Gibbs energy calculations confirm methanogenesis is exergonic in Guaymas sediments, with methylotrophic pathways consistently yielding more energy than the canonical hydrogenotrophic and acetoclastic pathways. Yet, metagenomic sequencing and cultivation attempts indicate that methanogens are present in low abundance. We find only one methyl-coenzyme M (mcrA) sequence within the entire sequencing dataset. Also, we identify a wide diversity of methyltransferases (mtaB, mttB), but only a few sequences phylogenetically cluster with methylotrophic methanogens. Our results suggest that the microbial methane in the Guaymas subsurface was produced over geologic time by relatively small methanogen populations, which have been variably influenced by thermal sediment alteration. Higher resolution metagenomic sampling may clarify the modern methanogen community. This study highlights the importance of using a multidisciplinary approach to capture microbial influences in dynamic, deep subsurface settings like Guaymas Basin.