Tropical peatlands are hotspots of methane (CH4) production but present high variation and emission uncertainties in the Amazon region. This is because the controlling factors of methane production in tropical peats are not yet well documented. Although inhibitory effects of nitrogen oxides (NOx) on methanogenic activity are known from pure culture studies, the role of NOx in the methane cycling of peatlands remains unexplored. Here, we investigated the CH4 content, soil geochemistry and microbial communities along 1-m-soil profiles and assessed the effects of soil NOx and nitrous oxide (N2O) on methanogenic abundance and activity in three peatlands of the Pastaza-Marañón foreland basin. The peatlands were distinct in pH, DOC, nitrate pore water concentrations, C/N ratios of shallow soils, redox potential, and 13C enrichment in dissolved inorganic carbon and CH4 pools, which are primarily contingent on H2-dependent methanogenesis. Molecular 16S rRNA and mcrA gene data revealed diverse and novel methanogens varying across sites. Importantly, we also observed a strong stratification in relative abundances of microbial groups involved in NOx cycling, along with a concordant stratification of methanogens. The higher relative abundance of ammonia-oxidizing archaea (Thaumarchaeota) in acidic oligotrophic peat than ammonia-oxidizing bacteria (Nitrospira) is noteworthy as putative sources of NOx. Experiments testing the interaction of NOx species and methanogenesis found that the latter showed differential sensitivity to nitrite (up to 85% reduction) and N2O (complete inhibition), which would act as an unaccounted CH4 control in these ecosystems. Overall, we present evidence of diverse peatlands likely differently affected by inhibitory effects of nitrogen species on methanogens as another contributor to variable CH4 fluxes.

The search for life on exoplanets is one of the grand scientific challenges of our time. The strategy to date has been to find (e.g., through transit surveys like Kepler) Earthlike exoplanets in their stars habitable zone, then use transmission spectroscopy to measure biosignature gases, especially oxygen, in the planets atmospheres (e.g., using JWST, the James Webb Space Telescope). Already there are more such planets than can be observed by JWST, and missions like the Transiting Exoplanet Survey Satellite and others will find more. A better understanding of the geochemical cycles relevant to biosignature gases is needed, to prioritize targets for costly follow-up observations and to help design future missions. We define a Detectability Index to quantify the likelihood that a biosignature gas could be assigned a biological vs. non-biological origin. We apply this index to the case of oxygen gas, O 2 , on Earth-like planets with varying water contents. We demonstrate that on Earth-like exoplanets with 0.2 weight percent (wt%) water (i.e., no exposed continents) a reduced flux of bioessential phosphorus limits the export of photosynthetically produced atmospheric O 2 to levels indistinguishable from geophysical production by photolysis of water plus hydrogen escape. Higher water contents >1wt% that lead to high-pressure ice mantles further slow phosphorus cycling. Paradoxically, the maximum water content allowing use of O 2 as a biosignature, 0.2wt%, is consistent with no water based on mass and radius. Thus, the utility of an O 2 biosignature likely requires the direct detection of both water and land on a planet.

Trace metals have been an important ingredient for life throughout Earth’s history. Here, we describe the genome-guided cultivation of a member of the elusive archaeal lineage Caldarchaeales (syn. Aigarchaeota), Wolframiiraptor gerlachensis, and its growth dependence on tungsten. A metagenome-assembled genome (MAG) of W. gerlachensis encodes putative tungsten membrane transport systems, as well as pathways for anaerobic oxidation of sugars probably mediated by tungsten-dependent ferredoxin oxidoreductases that are expressed during growth. Catalyzed reporter deposition-fluorescence in-situ hybridization (CARD-FISH) and nanoscale secondary ion mass spectrometry (nanoSIMS) show that W. gerlachensis preferentially assimilates xylose. Phylogenetic analyses of 78 high-quality Wolframiiraptoraceae MAGs from terrestrial and marine hydrothermal systems suggest that tungsten-associated enzymes were present in the last common ancestor of extant Wolframiiraptoraceae. Our observations imply a crucial role for tungsten-dependent metabolism in the origin and evolution of this lineage, and hint at a relic metabolic dependence on this trace metal in early anaerobic thermophiles.

Abstract. Chemodenitrification – the non-enzymatic process of nitrite reduction – may be an important sink for fixed nitrogen in tropical peatlands. Rates and products of chemodenitrification are dependent on O2, pH, Fe2+ concentration, and organic matter composition, which are variable across peat soils. Assessing abiotic reaction pathways is difficult because sterilization and inhibition agents can alter the availability of reactants by changing iron speciation and organic matter composition. We compared six commonly used soil sterilization techniques – γ irradiation, chloroform, autoclaving, and the use of three different chemical inhibitors (mercury, zinc, and azide) – for their compatibility with chemodenitrification assays for tropical peatland soils (organic-rich, low-pH soil from the eastern Amazon). Out of the six techniques, γ irradiation resulted in soil treatments with the lowest cell viability and denitrification activity and the least effect on pH, iron speciation, and organic matter composition. Nitrite depletion rates in γ-irradiated soils were highly similar to untreated (live) soils, whereas other sterilization techniques showed deviations. Chemodenitrification was a dominant process of nitrite consumption in tropical peatland soils assayed in this study. Nitrous oxide (N2O) is one possible product of chemodenitrification reactions. Abiotic N2O production was low to moderate (3 %–16 % of converted nitrite), and different sterilization techniques lead to significant variations on production rates due to inherent processes or potential artifacts. Our work represents the first methodological basis for testing the abiotic denitrification and N2O production potential in tropical peatland soil.

The preeminent source of biological methane on Earth is methyl coenzyme M reductase (Mcr)-dependent archaeal methanogenesis. A growing body of evidence suggests a diversity of archaea possess Mcr, although experimental validation of hypothesized methane metabolisms has been missing. Here, we provide evidence of a functional Mcr-based methanogenesis pathway in a novel member of the family Archaeoglobaceae, designated Methanoglobus nevadensis, which we enriched from a terrestrial hot spring on the polysaccharide xyloglucan. Our incubation assays demonstrate methane production that is highly sensitive to the Mcr inhibitor bromoethanesulfonate, stimulated by xyloglucan and xyloglucan-derived sugars, concomitant with the consumption of molecular hydrogen, and causing a deuterium fractionation in methane characteristic of hydrogenotrophic and methylotrophic methanogens. Combined with the recovery and analysis of a high-quality M. nevadensis metagenome-assembled genome encoding a divergent Mcr and diverse potential electron and carbon transfer pathways, our observations suggest methanogenesis in M. nevadensis occurs via Mcr and is fueled by the consumption of cross-fed byproducts of xyloglucan fermentation mediated by other community members. Phylogenetic analysis shows close affiliation of the M. nevadensis Mcr with those from Korarchaeota, Nezhaarchaeota, Verstraetearchaeota, and other Archaeoglobales that are divergent from well-characterized Mcr. We propose these archaea likely also use functional Mcr complexes to generate methane on the basis of our experimental validation in M. nevadensis. Thus, divergent Mcr-encoding archaea may be underestimated sources of biological methane in terrestrial and marine hydrothermal environments.