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Biochar was recently identified as an effective soil amendment for CH4 capture. Corresponding mechanisms are currently recognized to be from physical properties of biochar, providing a favorable growth environment for aerobic methanotrophs which perform aerobic methane (CH4) oxidation. However, our study shows that the chemical reactivity of biochar can also stimulate anaerobic oxidation of CH4 (AOM) by anaerobic methanotrophic archaea (ANME) of ANME-2d, which proposes another plausible mechanism for CH4 mitigation by biochar amendment in anaerobic environments. It was found that, by adding biochar as the sole electron acceptor in an anaerobic environment, CH4 was biologically oxidized, with CO2 production of 106.3 ± 5.1 μmol g–1 biochar. In contrast, limited CO2 production was observed with chemically reduced biochar amendment. This biological nature of the process was confirmed by mcr gene transcript abundance as well as sustained dominance of ANME-2d in the microbial community during microbial incubations with active biochar amendment. Combined FTIR and XPS analyses demonstrated that the redox activity of biochar is related to its oxygen-based functional groups. On the basis of microbial community evolution as well as intermediate production during incubation, different pathways in terms of direct or indirect interactions between ANME-2d and biochar were proposed for biochar-mediated AOM.

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