Identification of Methanogens and Controls on Methane Production in Incubations of Natural Methane Seep Sediments

Methane, the most abundant hydrocarbon in Earth's atmosphere, is produced in large quantities in sediments underlying the world's oceans. Very little of this methane makes it to surface sediments as it is consumed by Anaerobic Methanotrophs (ANME's) in consortia with Sulfate Reducing Bacteria (SRB). Less is known about which organisms are responsible for methane production in marine sediments, and whether that production is under thermodynamic control based on hydrogen concentrations. Although ANMEs have been found to be active in methanogenic sediments and incubations, it is currently unknown whether they are able to grow in methanogenic conditions. We demonstrated with bottle incubations of methane seep sediment taken from Cape Lookout Bight, NC, that hydrogen controls methane production. While sulfate was present the hydrogen concentration was maintained at below 2 nM. Only after the depletion of sulfate allowed hydrogen concentrations to rise above 5 nM did we see production of methane. The same sediments when spiked with methane gas demonstrated its complete removal while sulfate reduction occurred. Quantitative PCR shows that ANME-2 and ANME-1 increase in 16S copy number as methane increases. Total direct cell counts demonstrate a decline in cells with the decrease of sulfate until a recovery corresponding with production of methane. Our results strongly suggest that hydrogen concentrations influence what metabolic processes can occur in marine sediments, and that ANME-1 and ANME-2 are able to grow on the energy provided from methane production.