Molecular evidence for sediment nitrogen fixation in a temperate New England estuary

PeerJ. 2016 Jan 25:4:e1615. doi: 10.7717/peerj.1615. eCollection 2016.

Abstract

Primary production in coastal waters is generally nitrogen (N) limited with denitrification outpacing nitrogen fixation (N2-fixation). However, recent work suggests that we have potentially underestimated the importance of heterotrophic sediment N2-fixation in marine ecosystems. We used clone libraries to examine transcript diversity of nifH (a gene associated with N2-fixation) in sediments at three sites in a temperate New England estuary (Waquoit Bay, Massachusetts, USA) and compared our results to net sediment N2 fluxes previously measured at these sites. We observed nifH expression at all sites, including a site heavily impacted by anthropogenic N. At this N impacted site, we also observed mean net sediment N2-fixation, linking the geochemical rate measurement with nifH expression. This same site also had the lowest diversity (non-parametric Shannon = 2.75). At the two other sites, we also detected nifH transcripts, however, the mean N2 flux indicated net denitrification. These results suggest that N2-fixation and denitrification co-occur in these sediments. Of the unique sequences in this study, 67% were most closely related to uncultured bacteria from various marine environments, 17% to Cluster III, 15% to Cluster I, and only 1% to Cluster II. These data add to the growing body of literature that sediment heterotrophic N2-fixation, even under high inorganic nitrogen concentrations, may be an important yet overlooked source of N in coastal systems.

Keywords: Denitrification; Heterotrophic nitrogen fixation; Sediments; Sulfate-reducing bacteria; nifH; nifH diversity.

Grants and funding

This work was supported by Woods Hole Sea Grant and MIT Sea Grant. Additionally, this work was supported by the a Sloan Fellowship to RWF. SEN was funded by an NSF Postdoctoral Fellowship in Biology (1103692) The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.