Behaviours of methane metabolism and community dynamics of methane anaerobic oxidation microbes on carbonate rocks with long-term cultivation in cold seep environment

The anaerobic oxidation of methane (AOM) and sulfate reduction processes in cold seep environments can control methane emission sources and thus mitigate the pressure of increasing global greenhouse gas concentrations. The surfaces of carbonate rocks in cold seep host an abundance of microorganisms that participate in AOM reactions. Investigating the metabolism and conversion of methane by these microbes is instrumental in advancing the exploration and utilization of deep-sea methane energy. Previous studies primarily focused on in-situ investigations of microbial communities on carbonate rocks in cold seep environments, while the dynamic balance of carbonate mineralization caused by microbial community changes and the characteristics of methane consumption in carbonate samples remains unclear. In this study, we used methane as the sole carbon source, enriched and cultured carbonate samples under high pressure, and monitored the community dynamics regularly. The results demonstrated that methane consumption and metabolic pathways played a crucial role in influencing community succession and carbonate mineralization. AOM processes, coupled with sulfate reduction and nitrate reduction, which are dominated by ANME-2c, facilitate the precipitation of calcium carbonate. Conversely, the acetate production process, dominated by ANME-1, may hinder the efficiency of calcium carbonate mineralization. Additionally, the impact of bacterial groups in the enrichment process, through the production of extracellular enzymes, organic acid pathways, and the expression of carbonic anhydrase pathways on carbonate mineralization, should not be disregarded. These findings highlight the significance of diverse pathway methane metabolism in the dynamics of methane consumption, deep-sea microbial communities, carbonate kinetics, and marine biological carbon sequestration processes.