Wetlands constitute the main natural source of methane on Earth due to their high content of natural organic matter (NOM), but key drivers, such as electron acceptors, supporting methanotrophic activities in these habitats are poorly understood. We performed anoxic incubations using freshly collected sediment, along with water samples harvested from a tropical wetland, amended with 13 C-methane (0.67 atm) to test the capacity of its microbial community to perform anaerobic oxidation of methane (AOM) linked to the reduction of the humic fraction of its NOM. Collected evidence demonstrates that electron-accepting functional groups (e.g., quinones) present in NOM fueled AOM by serving as a terminal electron acceptor. Indeed, while sulfate reduction was the predominant process, accounting for up to 42.5% of the AOM activities, the microbial reduction of NOM concomitantly occurred. Furthermore, enrichment of wetland sediment with external NOM provided a complementary electron-accepting capacity, of which reduction accounted for ϳ100 nmol 13 CH 4 oxidized · cm Ϫ3 · day Ϫ1 . Spectroscopic evidence showed that quinone moieties were heterogeneously distributed in the wetland sediment, and their reduction occurred during the course of AOM. Moreover, an enrichment derived from wetland sediments performing AOM linked to NOM reduction stoichiometrically oxidized methane coupled to the reduction of the humic analogue anthraquinone-2,6-disulfonate. Microbial populations potentially involved in AOM coupled to microbial reduction of NOM were dominated by divergent biota from putative AOMassociated archaea. We estimate that this microbial process potentially contributes to the suppression of up to 114 teragrams (Tg) of CH 4 · year Ϫ1 in coastal wetlands and more than 1,300 Tg · year Ϫ1 , considering the global wetland area. IMPORTANCEThe identification of key processes governing methane emissions from natural systems is of major importance considering the global warming effects triggered by this greenhouse gas. Anaerobic oxidation of methane (AOM) coupled to the microbial reduction of distinct electron acceptors plays a pivotal role in mitigating methane emissions from ecosystems. Given their high organic content, wetlands constitute the largest natural source of atmospheric methane. Nevertheless, processes controlling methane emissions in these environments are poorly understood. Here, we provide tracer analysis with 13 CH 4 and spectroscopic evidence revealing that AOM linked to the microbial reduction of redox functional groups in natural organic matter (NOM) prevails in a tropical wetland. We suggest that microbial reduction of NOM may largely contribute to the suppression of methane emissions from

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Availability of fixed nitrogen is a pivotal driver on primary productivity in the oceans, thus the identification of key processes triggering nitrogen losses from these ecosystems is of major importance as they affect ecosystems function and consequently global biogeochemical cycles. Denitrification and anaerobic ammonium oxidation coupled to nitrite reduction (Anammox) are the only identified marine sinks for fixed nitrogen. The present study provides evidence indicating that anaerobic ammonium oxidation coupled to the reduction of sulfate, the most abundant electron acceptor present in the oceans, prevails in marine sediments. Tracer analysis with N-ammonium revealed that this microbial process, here introduced as Sulfammox, accounts for up to 5 μgN produced g day in sediments collected from the eastern tropical North Pacific coast. Raman and X-ray diffraction spectroscopies revealed that elemental sulfur and sphalerite (ZnFeS) were produced, besides free sulfide, during the course of Sulfammox. Anaerobic ammonium oxidation linked to Fe(III) reduction (Feammox) was also observed in the same marine sediments accounting for up to 2 μg N produced g day. Taxonomic characterization, based on 16S rRNA gene sequencing, of marine sediments performing the Sulfammox and Feammox processes revealed the microbial members potentially involved. These novel nitrogen sinks may significantly fuel nitrogen loss in marine environments. These findings suggest that the interconnections among the oceanic biogeochemical cycles of N, S and Fe are much more complex than previously considered.

Identification of microbial processes driving the loss of nitrogen from the oceans is of paramount relevance as these processes affect primary productivity in these ecosystems, which ultimately affects global biogeochemical cycles. Denitrification and anammox (anaerobic ammonium oxidation coupled to nitrite reduction) are the only identified processes so far that lead to nitrogen loss in marine environments. Here we provide stoichiometric and spectroscopic evidence, as well as tracer analysis with [15N]­ammonium, revealing that anaerobic ammonium oxidation linked to the microbial reduction of natural organic matter (NOM) fuels nitrogen loss in marine sediments from the eastern tropical North Pacific coast. Tracer analysis revealed that the NOM-dependent anammox process was responsible for producing ∼1.5 μg of 15N2 (g of sediment)−1 day–1 after incubation for 27 days in sediment incubations amended with Pahokee peat, while intrinsic NOM present in the sediment promoted the production of ∼0.4 μg of 15N2 (g of sediment)−1 day–1. Taxonomic characterization, based on 16S rRNA gene sequencing, of the biota present in marine sediments performing the NOM-dependent anammox process revealed several microbial members are potentially involved. The most predominant bacterial phylotypes detected were associated with Phycisphaeraceae, Actinomarinales, Acidiferrobacteraceae, and Rhodobacteraceae, while Nitrosopumilaceae was the only archaeal family whose level clearly increased during the course of NOM-dependent anammox. This is a novel pathway interconnecting the oceanic biogeochemical cycles of N and C, which may significantly propel nitrogen fluxes in organic-rich, coastal marine sediments.

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