Different responses of nitrite- and nitrate-dependent anaerobic methanotrophs to increasing nitrogen loading in a freshwater reservoir

Abstract

Nitrite (NO2−)- and nitrate (NO3−)-dependent anaerobic oxidation of methane (AOM) are two new additions in microbial methane cycle, which potentially act as important methane sinks in freshwater aquatic systems. Here, we investigated spatial variations of community composition, abundance and potential activity of NO2−- and NO3−-dependent anaerobic methanotrophs in the sediment of Jiulonghu Reservoir (Zhejiang Province, China), a freshwater reservoir having a gradient of increasing nitrogen loading from upstream to downstream regions. High-throughput sequencing of total bacterial and archaeal 16S rRNA genes showed the cooccurrence of Candidatus Methylomirabilis oxyfera (M. oxyfera)-like and Candidatus Methanoperedens nitroreducens (M. nitroreducens)-like anaerobic methanotrophs in the examined reservoir sediments. The community structures of these methanotrophs differed substantially between the sediments of upstream and downstream regions. Quantitative PCR suggested higher M. oxyfera-like bacterial abundance in the downstream (8.6 × 107 to 2.8 × 108 copies g−1 dry sediment) than upstream sediments (2.4 × 107 to 3.5 × 107 copies g−1 dry sediment), but there was no obvious difference in M. nitroreducens-like archaeal abundance between these sediments (3.7 × 105 to 4.8 × 105 copies g−1 dry sediment). The 13CH4 tracer experiments suggested the occurrence of NO2−- and NO3−-dependent AOM activities, and their rates were 4.7–14.1 and 0.8–2.6 nmol CO2 g−1 (dry sediment) d−1, respectively. Further, the rates of NO2−-dependent AOM in downstream sediment were significantly higher than those in upstream sediment. The NO3− concentration was the key factor affecting the spatial variations of abundance and activity of NO2−-dependent anaerobic methanotrophs. Overall, our results showed different responses of NO2−- and NO3−-dependent anaerobic methanotrophs to increasing nitrogen loading in a freshwater reservoir.