Long-term N-addition alters the community structure of functionally important N-cycling soil microorganisms across global grasslands

Beat Frey, Barbara Moser, Bjorn Tytgat, Stephan Zimmermann, Juan Alberti, Lori A. Biederman, Elizabeth T. Borer, Arthur A.D. Broadbent, Maria C. Caldeira, Kendi F. Davies, Nico Eisenhauer, Anu Eskelinen, Philip A. Fay, Frank Hagedorn, Yann Hautier, Andrew S. MacDougall, Rebecca L. McCulley, Joslin L. Moore, Maximilian Nepel, Sally A. PowerEric W. Seabloom, Eduardo Vázquez, Risto Virtanen, Laura Yahdjian, Anita C. Risch

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


Anthropogenic nitrogen (N) input is known to alter the soil microbiome, but how N enrichment influences the abundance, alpha-diversity and community structure of N-cycling functional microbial communities in grasslands remains poorly understood. Here, we collected soils from plant communities subjected to up to 9 years of annual N-addition (10 g N m−2 per year using urea as a N-source) and from unfertilized plots (control) in 30 grasslands worldwide spanning a large range of climatic and soil conditions. We focused on three key microbial groups responsible for two essential processes of the global N cycle: N2 fixation (soil diazotrophs) and nitrification (AOA: ammonia-oxidizing archaea and AOB: ammonia-oxidizing bacteria). We targeted soil diazotrophs, AOA and AOB using Illumina MiSeq sequencing and measured the abundance (gene copy numbers) using quantitative PCR. N-addition shifted the structure of the diazotrophic communities, although their alpha-diversity and abundance were not affected. AOA and AOB responded differently to N-addition. The abundance and alpha-diversity of AOB increased, and their community structure shifted with N-addition. In contrast, AOA were not affected by N-addition. AOA abundance outnumbered AOB in control plots under conditions of low N availability, whereas N-addition favoured copiotrophic AOB. Overall, N-addition showed a low impact on soil diazotrophs and AOA while effects for AOB communities were considerable. These results reveal that long-term N-addition has important ecological implications for key microbial groups involved in two critical soil N-cycling processes. Increased AOB abundance and community shifts following N-addition may change soil N-cycling, as larger population sizes may promote higher rates of ammonia oxidation and subsequently increase N loss via gaseous and soil N-leaching. These findings bring us a step closer to predicting the responses and feedbacks of microbial-mediated N-cycling processes to long-term anthropogenic N-addition in grasslands.

Original languageEnglish (US)
Article number108887
JournalSoil Biology and Biochemistry
StatePublished - Jan 2023

Bibliographical note

Funding Information:
This work was conducted within the Nutrient Network ( http://www.nutnet.org ) experiment, which is funded at the site scale by individual researchers. The N-cycling microbial communities add-on study was funded by an internal competitive WSL grant (PSP 5233.00176.001.01) to B.F. and A.C.R. Coordination and data management have been supported by funding from the National Science Foundation Research Coordination Network (NSF-DEB-1042132) to E.T.B. and E.W.S., the Long-Term Ecological Research (LTER) programme (NSF-DEB-1234162), and the Institute on the Environment at the University of Minnesota (DG-0001-13). We also thank the Minnesota Supercomputer Institute for hosting project data, and the Institute on the Environment for hosting NutNet meetings. We are grateful to B. Stierli, S. Baumgartner and A. Dharmarajah for their help with sample processing and analyses. We also thank M. Dawes for her valuable contribution to the editing of this article.

Publisher Copyright:
© 2022 The Authors


  • Ammonia oxidizer
  • Biogeography
  • Diazotroph
  • Grassland
  • N-Fertilization
  • N-cycling microbial community
  • N-fixing bacteria
  • Nutrient network (NutNet)
  • Urea
  • nifH


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