Deciphering cryptic methane cycling: Coupling of methylotrophic methanogenesis and anaerobic oxidation of methane in hypersaline coastal wetland sediment

Methanogenesis has recently been shown to fuel anaerobic oxidation of methane (AOM) within the sulfate-reducing zone of marine sediments, coining the term "cryptic methane cycle". Here we present research on the relationship between methanogenesis and AOM in a shallow hypersaline pool (∼130 PSU) within a southern California coastal wetland. Sediment (top 20 cm) was subjected to geochemical analyses, in-vitro slurry experiments, and radiotracer incubations using 35S-SO42−, 14C-mono-methylamine, and 14C-CH4, to study sulfate reduction, methylotrophic methanogenesis, and AOM. An adapted radioisotope method was used to follow cryptic methane cycling in 14C-mono-methylamine labeling incubations with increasing incubation times (1 hour to three weeks). Results showed peaks in AOM (max 13 nmol cm−3 d−1) and sulfate reduction activity (max 728 nmol cm−3 d−1) within the top 6 cm. Below 6 cm, AOM activity continued (max 15 nmol cm−3 d−1), while sulfate reduction was absent despite 67 mM sulfate, suggesting AOM was coupled to the reduction of iron. Methane concentrations were low (<50 nM) throughout the sediment. Batch sediment slurry incubations with methylated substrates (mono-methylamine and methanol) stimulated methanogenesis, pointing to the presence of methylotrophic methanogens. Incubations with 14C-mono-methylamine revealed the simultaneous activity of methanogenesis and coupled AOM through the stepwise transfer of 14C from mono-methylamine to CO2 via methane. Our results suggest that AOM is a crucial process in coastal wetland sediments to prevent the buildup of methane in the sulfate-reducing zone. We propose that cryptic methane cycling has been largely overlooked in coastal wetlands resulting in incomplete understanding of carbon cycling in this environment.