Skip to main content
SpringerLink
Log in

Diazotrophic bacterial community variability in a subtropical deep reservoir is correlated with seasonal changes in nitrogen

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Nitrogen-fixing microorganisms (diazotrophs) play important roles in aquatic biogeochemistry and ecosystem functioning. However, little is known about the spatiotemporal variation of diazotrophic microbial communities in deep subtropical reservoirs. In this study, denaturing gradient gel electrophoresis (DGGE), clone libraries, quantitative PCR, and quantitative reverse transcription (RT)-PCR were used together to examine the vertical and seasonal patterns of diazotrophic microbial communities based on nitrogenase (nifH) gene sequences in the Dongzhen Reservoir, China, across time (every 3 months for 1 year) and space (five different water depths). In general, the numbers of DGGE bands increased with water depth during the stratification seasons (spring, summer, and autumn), with the clone-library-based operational taxonomic unit (OTU) number and nifH gene diversity being highest in autumn (6 OTUs at depth 0 m; 15 OTUs at 33 m) and winter (12 OTUs at 0 m, 13 OTUs at 33 m) but decreasing drastically in spring (2 OTUs at 0 m, 3 OTUs at 33 m) and summer (3 OTUs at 0 m, 2 OTUs at 33 m). The nifH gene abundance was lowest in the water mixing season (winter average, 5.17 × 107 copies/L) but increased in the three other seasons (9.03 × 109 copies/L). Cyanobacteria (dominated by filamentous thermophilic cyanobacteria and Cylindrospermopsis raciborskii) were the most dominant diazotrophic group at all depths and seasons, while both alphaproteobacteria and gammaproteobacteria were co-dominant in the bottom waters in autumn and winter. The distinct seasonal and spatial patterns in diazotrophic communities were significantly related to total nitrogen (TN) and ammonium nitrogen (NH4-N) in the reservoir (P < 0.01). Further, TN showed a significant positive correlation with nifH RNA copy number (P < 0.05) and DGGE band number (P < 0.01), whereas the NH4-N was negatively correlated with nifH DNA copy number (P < 0.01) and positively with both RNA/DNA ratio (P < 0.01) and DGGE band number (P < 0.01). Our data indicated that water stratification, mixing, and nitrogen might drive the diazotrophic community structure and activity in complex ways, thereby influencing the aquatic nitrogen cycle. Therefore, adaptive reservoir management strategies should carefully consider the effects of water stratification for protecting drinking water quality and for controlling the potential for diazotrophic cyanobacteria blooms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bagwell CE, Lovell CR (2000) Microdiversity of culturable diazotrophs from the rhizoplanes of the salt marsh grasses Spartina alterniflora and Juncus roemerianus. Microb Ecol 39:128–136

    Article  CAS  Google Scholar 

  • Becker V, Huszar VLM, Crossetti LO (2009) Responses of phytoplankton functional groups to the mixing regime in a deep subtropical reservoir. Hydrobiologia 628:137–151

    Article  Google Scholar 

  • Becker V, Huszar VLM, Naselli-Flores L, Padiak J (2008) Phytoplankton equilibrium phases during thermal stratification in a deep subtropical reservoir. Freshw Biol 53:952–963

    Article  Google Scholar 

  • Braun ST, Proctor LM, Zani S, Mellon MT, Zehr JP (1999) Molecular evidence for zooplankton-associated nitrogen-fixing anaerobes based on amplification of the nifH gene. FEMS Microbiol Ecol 28:273–279

    Article  CAS  Google Scholar 

  • Bustin SA, Benes V, Garson JA, Hellemans J, Huqqett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622

    Article  CAS  Google Scholar 

  • Canfield DE, Glazer AN, Falkowski PG (2010) The evolution and future of Earth’s nitrogen cycle. Science 330:192–196

    Article  CAS  Google Scholar 

  • Capone DG, Zehr JP, Paerl HW, Bergman B, Carpenter EJ (1997) Trichodesmium, a globally significant marine cyanobacterium. Science 276:1221–1229

    Article  CAS  Google Scholar 

  • Chao A (1984) Nonparametric estimation of the number of classes in a population. Scand J Stat 11:265–270

    Google Scholar 

  • Chao A, Lee SM (1992) Estimating the number of classes via sample coverage. J Am Stat Assoc 87:210–217

    Article  Google Scholar 

  • Clark KR, Gorley RN (2001) PRIMER v5: user manual/tutorial. PRIMER-E, Plymouth, UK

    Google Scholar 

  • Dixon R, Kahn D (2004) Genetic regulation of biological nitrogen fixation. Nat Rev Microbiol 2:621–631

    Article  CAS  Google Scholar 

  • Falkowski PG (1983) Enzymology of nitrogen assimilation. In: Carpenter EJ, Capone DG (eds) Nitrogen in the marine environment. Academic press, New York, USA, pp 839–868

    Chapter  Google Scholar 

  • Farnelid H, Andersson AF, Bertilsson S, Al-Soud WA, Hansen LH, Sørensen S, Steward GF, Hagström Å, Riemann L (2011) Nitrogenase gene amplicons from global marine surface waters are dominated by genes of non-cyanobacteria. PLoS ONE 6:e19223

    Article  CAS  Google Scholar 

  • Farnelid H, Bentzon-Tilia M, Andersson AF, Bertilsson S, Labrebz M, Jürgens K, Riemann L (2013) Active nitrogen-fixing heterotrophic bacteria at and below the chemocline of the central Baltic Sea. ISME J 7:1413–1423

    Article  CAS  Google Scholar 

  • Flores E, Herrero A (1994) Assimilatory nitrogen metabolism and its regulation. In: Bryant DA (ed) The molecular biology of cyanobacteria. Kluwer Academic Publishers, the Netherlands, pp 487–517

    Chapter  Google Scholar 

  • Flores E, Muro-Pastor AM, Herrero A (1999) Cyanobacterial nitrogen assimilation genes and NtcA-dependent control of gene expression. In: Peschek GA, Löffelhardt W, Schmetterer G (eds) The phototrophic prokaryotes. Plenum Publishing Corporation, New York, USA, pp 463–477

    Chapter  Google Scholar 

  • Gaby JC, Buckley DH (2011) A global census of nitrogenase diversity. Environ Microbiol 13:1790–1799

    Article  CAS  Google Scholar 

  • Gaby JC, Buckley DH (2012) A comprehensive evaluation of PCR primers to amplify the nifH gene of nitrogenase. PLoS ONE 7:e42149

    Article  CAS  Google Scholar 

  • Haegeman B, Hamelin J, Moriarty J, Neal P, Dushoff J, Weitz JS (2013) Robust estimation of microbial diversity in theory and in practice. ISME J 7:1092–1101

    Article  Google Scholar 

  • Hamersley MR, Turk KA, Leinweber A, Gruber N, Zehr JP, Gunderson T, Capone DG (2011) Nitrogen fixation within the water column associated with two hypoxic basins within the Southern California Bight. Aquat Microb Ecol 63:193–205

    Article  Google Scholar 

  • Herrero A, Muro-Pastor AM, Flores E (2001) Nitrogen control in cyanobacteria. J Bacteriol 183:411–425

    Article  CAS  Google Scholar 

  • Hulot FD, Huisman J (2004) Allelopathic interactions between phytoplankton species: the roles of heterotrophic bacteria and mixing intensity. Limnol Oceanogr 49:1424–1434

    Article  Google Scholar 

  • Ionescu D, Oren A, Levitan O, Hindiyeh M, Malkami H, Berman-Frank L (2009) The cyanobacterial community of the Zerka Ma'in hot springs, Jordan: morphological and molecular diversity and nitrogen fixation. Algol Stud 130:109–124

    Article  CAS  Google Scholar 

  • Jayakumar A, Al-Rshaidat MMD, Ward BB, Mulholland MR (2012) Diversity, distribution, and expression of diazotroph nifH genes in oxygen-deficient waters of the Arabian Sea. FEMS Microbiol Ecol 82:597–606

    Article  CAS  Google Scholar 

  • Kanemoto RH, Ludden PW (1984) Effect of ammonia, darkness, and phenazine methosulfate on whole-cell nitrogenase activity and Fe protein modification in Rhodospirillum rubrum. J Bacteriol 158:713–720

    CAS  Google Scholar 

  • Kerr B, Riley MA, Feldman MW, Bohannan BJ (2002) Local dispersal promotes biodiversity in a real-life game of rock-paper-scissors. Nature 418:171–174

    Article  CAS  Google Scholar 

  • Klugkist J, Haaker H (1984) Inhibition of nitrogenase activity by ammonium chloride in Azotobacter vinelandii. J Bacteriol 157:148–151

    CAS  Google Scholar 

  • Knapp AN (2012) The sensitivity of marine N2 fixation to dissolved inorganic nitrogen. Front Microbiol 3:374

    CAS  Google Scholar 

  • Kumar K, Mella-Herrera RA, Golden JW (2010) Cyanobacterial heterocysts. Cold Spring Harb Perspect Biol 2:a000315

    Article  Google Scholar 

  • Langlois RJ, Hummer D, LaRoche J (2008) Abundances and distributions of the dominant nifH phylotypes in the Northern Atlantic Ocean. Appl Environ Microbiol 74:1922–1931

    Article  CAS  Google Scholar 

  • Liu LM, Yang J, Zhang YY (2011) Genetic diversity patterns of microbial communities in a subtropical riverine ecosystem (Jiulong River, southeast China). Hydrobiologia 678:113–125

    Article  CAS  Google Scholar 

  • Lv H, Yang J, Liu LM (2013) Temporal pattern prevails over spatial variability in phytoplankton communities from a subtropical water supply reservoir. Oceanol Hydrobiol Stud 42:420–430

    Article  CAS  Google Scholar 

  • Lv H, Yang J, Liu LM, Yu XQ, Yu Z, Chiang P (2014) Temperature and nutrients are significant drivers of seasonal shift in phytoplankton community from a drinking water reservoir, subtropical China. Environ Sci Pollut Res 21:5917–5928

    Article  CAS  Google Scholar 

  • Man-Aharonovich D, Kress N, Zeev EB, Berman-Frank, Béjà O (2007) Molecular ecology of nifH genes and transcripts in the eastern Mediterranean Sea. Environ Microbiol 9:2354–2363

    Article  CAS  Google Scholar 

  • McGlathery KJ, Risgaard-Petersen N, Christensen PB (1998) Temporal and spatial variation in nitrogen fixation activity in the eelgrass Zostera marina rhizosphere. Mar Ecol Prog Ser 168:245–258

    Article  CAS  Google Scholar 

  • Ohki K, Zehr JP, Falkowski PG, Fujita Y (1991) Regulation of nitrogen-fixation by different nitrogen-sources in the marine nonheterocystous cyanobacterium Trichodesmium sp. NIBB1067. Arch Microbiol 156:335–337

    Article  CAS  Google Scholar 

  • Olson JB, Steppe TF, Litaker RW, Paerl HW (1998) N2-fixing microbial consortia associated with the ice cover of Lake Bonney, Antarctica. Microb Ecol 36:231–238

    Article  CAS  Google Scholar 

  • Patriquin D (1978) Nitrogen fixation (acetylene reduction) associated with cord grass, Spartina alterniflora Loisel. In: Granhall U (ed) Environmental role of nitrogen-fixing blue-green algae and asymbiotic bacteria. Ecological Bulletin, Stockholm, Sweden, pp 20–27

    Google Scholar 

  • Reynolds CS (2006) The ecology of phytoplankton (ecology, biodiversity and conservation). Cambridge University Press, Cambridge, UK

    Google Scholar 

  • Riemann L, Farnelid H, Steward GF (2010) Nitrogenase genes in non-cyanobacterial plankton: prevalence, diversity and regulation in marine waters. Aquat Microb Ecol 61:235–247

    Article  Google Scholar 

  • Schloss PD, Westcott SL, Ryabin T, Hall JP, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, Sahl JW, Stres B, Thallinger GG, Van Horn DJ, Weber CF (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541

    Article  CAS  Google Scholar 

  • Shade A, Jones SE, McMahon KD (2008) The influence of habitat heterogeneity on freshwater bacterial community composition and dynamics. Environ Microbiol 10:1057–1067

    Article  CAS  Google Scholar 

  • Short SM, Zehr JP (2007) Nitrogen gene expression in the Chesapeake Bay Estuary. Environ Microbiol 9:1591–1596

    Article  CAS  Google Scholar 

  • Staal M, Meysman FJR, Stal LJ (2003) Temperature excludes N2-fixing heterocystous cyanobacteria in the tropical oceans. Nature 425:504–507

    Article  CAS  Google Scholar 

  • Ter Braak CJF, Smilauer P (2002) CANOCO Reference Manual and CanoDraw for Windows User's Guide: Software for Canonical Community Ordination (version 4.5). Microcomputer Power: Ithac, New York, USA

  • Turk KA, Rees AP, Zehr JP, Pereira N, Swift P, Shelley R, Lohan M, Woodward EMS, Gilbert J (2011) Nitrogen fixation and nitrogenase (nifH) expression in tropical waters of the eastern North Atlantic. ISME J 5:1201–1212

    Article  CAS  Google Scholar 

  • Walsby AE (1985) The permeability of heterocysts to the gases nitrogen and oxygen. Proc R Soc Lond B 226:345–366

    Article  CAS  Google Scholar 

  • Walsby AE, Utkilen HC, Johnsen IJ (1983) Buoyancy changes of a red coloured Oscillatoria agardhii in Lake Gjersjøen, Norway. Arch Hydrobiol 97:18–38

    Google Scholar 

  • Wang S, Qian X, Han BP, Wang QH, Ding ZF (2011) Physical limnology of a typical subtropical reservoir in south China. Lake Reserv Manag 27:149–161

    Article  Google Scholar 

  • Wolk CP, Ernst A, Elhai J (1994) Heterocyst metabolism and development. In: Bryant DA (ed) The molecular biology of cyanobacteria. Kluwer Academic Publishers, Dordrecht, the Netherlands, pp 769–823

    Chapter  Google Scholar 

  • Yang J, Yu XQ, Liu LM, Zhang WJ, Guo P (2012) Algae community and trophic state of subtropical reservoirs in southeast Fujian, China. Environ Sci Pollut Res 19:1432–1442

    Article  CAS  Google Scholar 

  • Yu Z, Yang J, Stefano A, Yu XQ, Liu LM (2014a) Effects of water stratification and mixing on microbial community structure in a subtropical deep reservoir. Sci Rep 4:5821

    CAS  Google Scholar 

  • Yu Z, Yang J, Zhou J, Yu XQ, Liu LM, Lv H (2014b) Water stratification affects the microeukaryotic community in a subtropical deep reservoir. J Eukaryot Microbiol 61:126–133

    Article  Google Scholar 

  • Yu Z, Zhou J, Yang J, Yu XQ, Liu LM (2014c) Vertical distribution of diazotrophic bacterial community associated with temperature and oxygen gradients in a subtropical reservoir. Hydrobiologia 41:69–77

    Article  CAS  Google Scholar 

  • Zehr JP (2011) Nitrogen fixation by marine cyanobacteria. Trends Microbiol 19:162–173

    Article  CAS  Google Scholar 

  • Zehr JP, Church MJ, Moisander PH (2006) Diversity, distribution and biogeochemical significance of nitrogen-fixing microorganisms in anoxic and suboxic ocean environments. In: Neretin LN (ed) Past and present water column anoxia. Springer, Dordrecht, the Netherlands, pp 337–369

    Chapter  Google Scholar 

  • Zehr JP, Jenkins BD, Short SM, Steward GF (2003) Nitrogenase gene diversity and microbial community structure: a cross-system comparison. Environ Microbiol 5:539–554

    Article  CAS  Google Scholar 

Download references

Acknowledgments

David M Wilkinson and two reviewers provided valuable comments that improved this manuscript. This research was supported by the National Basic Research Program of China (2012CB956103), the National Natural Science Foundation of China (31370471, 31172114, and U1133601), and the Natural Science Foundation for Distinguished Young Scholars of Fujian Province (2012 J06009).

Compliance with ethical standards

No specific permissions were required for these activities. Informed consent was obtained from all participants, and this field study did not involve endangered or protected species.

Author information

Authors and Affiliations

  1. Aquatic EcoHealth Group, Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, 361021, China

    Lina Wang, Zheng Yu, Jun Yang & Jing Zhou

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Lina Wang

  3. School of Environmental Studies, China University of Geosciences (Wuhan), Wuhan, 430074, China

    Jing Zhou

Authors
  1. Lina Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zheng Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun Yang.

Additional information

Responsible editor: Robert Duran

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOC 656 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, L., Yu, Z., Yang, J. et al. Diazotrophic bacterial community variability in a subtropical deep reservoir is correlated with seasonal changes in nitrogen. Environ Sci Pollut Res 22, 19695–19705 (2015). https://doi.org/10.1007/s11356-015-5144-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-015-5144-9

Keywords

Search

Navigation