Microbial community synergy drives metal-dependent anaerobic methane oxidation under simulated deep-sea high-pressure conditions

Abstract

The anaerobic oxidation of methane linked to metal oxide reduction (metal-dependent AOM) plays a crucial role in regulating methane fluxes within deep-sea sedimentary environments. While metal-dependent AOM has been reported, current understanding is primarily derived from in situ investigations and enrichment experiments conducted under atmospheric pressure, leaving its applicability to the high-pressure, low-temperature deep-sea environment uncertain. In this study, we established long-term enrichment cultures of microbial communities from South China Sea cold seep sediments (1392 m) with manganese/iron oxides as electron acceptors under simulated in situ pressure-temperature conditions (12 MPa, 4 °C). Our results demonstrate significant methane consumption (approximately 30%) accompanied by the accumulation of Mn (II) and Fe (II) over 150 d. Microbial community analysis revealed that anaerobic methanotrophic archaea (ANME-1) dominated the archaeal community. Their predicted extracellularly secreted multi-heme cytochromes (MHCs) may facilitate direct electron transfer under high pressure. Metagenomic analysis further revealed a diversity of methane metabolic pathways within the community. Horizontal gene transfer may have facilitated microbial electron exchange, while ANME archaea engaged in cross-feeding with bacterial groups via metabolites such as acetate and amino acids. This study underscores the critical role of microbial community synergy and adaptive evolution in regulating methane oxidation under simulated deep-sea conditions, providing new insights into the marine carbon cycle.

Graphical abstract

Keywords

Anaerobic methane oxidation / Cold seep / methanotrophs / Mn/Fe coupling / Microbial interactions / Horizontal gene transfer / Carbon sequestration

Highlight

	● Simulated deep-sea pressure/temperature enables sustained methane-metal AOM.
	● Metagenomics reveals diverse methane pathways and horizontal gene transfer.
	● ANME-1 archaea dominate and possess putative extracellular multi-heme cytochromes.
	● Microbial synergy and cross-feeding drive methane oxidation in deep-sea simulation.

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