Abstract

Denitrification is an important pathway for nitrogen removal from aquatic systems and this could benefit water quality. However, little is known about the denitrifier community composition and key steps of denitrification in the freshwater environments, and whether different bacteria have a role in multiple processes of denitrification reduction. In this study, quantitative PCR, quantitative RT-PCR, clone library and 454 pyrosequencing were used together to investigate the bacterial and denitrifier community in a subtropical deep reservoir during the strongly stratified period. Our results indicated that the narG gene recorded the highest abundance among the denitrifying genes (2.76×109 copies L−1 for DNA and 4.19×108 copies L−1 for RNA), and the lowest value was nosZ gene (7.56×105 copies L−1 for DNA and undetected for RNA). The RNA: DNA ratios indicated that narG gene was the most active denitrifying gene in the oxygen minimum zone of Dongzhen Reservoir. Further, α-, β- and γ- Proteobacteria were the overwhelmingly dominant classes of denitrifier communities. Each functional gene had its own dominant groups which were different at the genus level: the narG gene was dominated by Albidiferax, while nirS gene was dominated by Dechloromonas. The main OTU of nirK gene was Rhodopseudomonas palustris, but for norB and nosZ genes, they were Bacillus and Bradyrhizobium, respectively. These results contribute to the understanding of linkages between denitrifier community, function and how they work together to complete the denitrification process. Studies on denitrifier community and activity may be useful in managing stratified reservoirs for the ecosystem services and aiding in constructing nitrogen budgets.

Highlights

  • IntroductionNitrogen (N), the fourth most abundant element (after oxygen, carbon and hydrogen) in microorganisms, is essential for the synthesis of nucleic acids and proteins [1]

  • Nitrogen (N), the fourth most abundant element in microorganisms, is essential for the synthesis of nucleic acids and proteins [1]

  • Based on the 99 (16 S rRNA), 64, 124, 69, 43 and 53 sequenced clones, 71, 31, 93, 24, 17 and 22 operational taxonomic unit (OTU) were obtained for each gene at the 97% similarity level, respectively (Figure 2A)

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Summary

Introduction

Nitrogen (N), the fourth most abundant element (after oxygen, carbon and hydrogen) in microorganisms, is essential for the synthesis of nucleic acids and proteins [1]. It is commonly found as amine or amide groups in organic matter but is readily oxidized or reduced and has an additional significance in aquatic systems as both an electron acceptor and a donor for energy metabolism [2]. Anthropogenic influences on the biogeochemistry of nitrogen for instance, combustion of fossil fuels, production of nitrogen fertilizers, cultivation of nitrogen-fixing legumes, and other actions have resulted in major changes in the earth’s nitrogen cycle [5]. Whilst the production and industrial use of artificial nitrogen fertilizers worldwide has enabled humankind to greatly increase food production, it has led to a host of environmental problems, especially potential eutrophication of freshwater and coastal ecosystems [6]

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