Nitrite Oxidizer Activity and Community Are More Responsive Than Their Abundance to Ammonium-Based Fertilizer in an Agricultural Soil

Autotrophic nitrification is mediated by ammonia oxidizing bacteria (AOB) or ammonia oxidizing archaea (AOA) and nitrite oxidizing bacteria (NOB). Mounting studies have examined the impact of nitrogen (N) fertilization on the dynamic and diversity of AOA and AOB, while we have limited information on the response of the activity, abundance, and diversity of NOB to N fertilization. We investigated the influence of organic and inorganic N fertilizers on soil NOB in silage corn field plots that received contrasting nitrogen (N) treatments: control (no additional N), ammonium sulfate (AS 100 and 200 kg N ha−1), and compost (200 kg N ha−1). Nitrifying community was examined using a universal marker (16S rRNA gene), functional gene markers (AOB amoA and Nitrospira nxrB), and metagenomics. The overall nitrifying community was not altered after the first fertilization but was significantly shifted by 4-year repeated application of ammonium fertilizers. Nitrospira were the dominant NOB (>99.7%) in our agricultural soil. Both community compositions of AOB and Nitrospira were significantly changed by ammonium fertilizers but not by compost after 4 years of repeated applications. All nitrifiers, including comammox, were recovered in soil metagenomes based on a gene-targeted assembly, but their sequence counts were very low. Although N treatment did not affect the abundance of Nitrospira nxrB determined by real-time quantitative PCR, ammonium fertilizers significantly promoted rates of potential nitrite oxidation determined at 0.15 mM nitrite in soil slurries. Understanding the response of both ammonia oxidizers and nitrite oxidizers to N fertilization may initiate or improve strategies for mitigating potential environmental impacts of nitrate production in agricultural ecosystems.

[1]  M. Firestone,et al.  Mechanisms for soil moisture effects on activity of nitrifying bacteria , 1995, Applied and environmental microbiology.

[2]  W. Liesack,et al.  The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations , 1997, Applied and environmental microbiology.

[3]  M. Wagner,et al.  Phylogeny of All Recognized Species of Ammonia Oxidizers Based on Comparative 16S rRNA and amoA Sequence Analysis: Implications for Molecular Diversity Surveys , 2000, Applied and Environmental Microbiology.

[4]  M. Klotz,et al.  Diversity of ammonia monooxygenase operon in autotrophic ammonia-oxidizing bacteria , 2002, Archives of Microbiology.

[5]  J. Prosser,et al.  Links between ammonia oxidizer species composition, functional diversity and nitrification kinetics in grassland soils. , 2005, Environmental microbiology.

[6]  T. Urich,et al.  Archaea predominate among ammonia-oxidizing prokaryotes in soils , 2006, Nature.

[7]  X. Le Roux,et al.  Decline of soil microbial diversity does not influence the resistance and resilience of key soil microbial functional groups following a model disturbance. , 2007, Environmental microbiology.

[8]  F. Poly,et al.  First exploration of Nitrobacter diversity in soils by a PCR cloning-sequencing approach targeting functional gene nxrA. , 2008, FEMS microbiology ecology.

[9]  X. Le Roux,et al.  Development and application of a PCR-denaturing gradient gel electrophoresis tool to study the diversity of Nitrobacter-like nxrA sequences in soil. , 2008, FEMS microbiology ecology.

[10]  Ning Ma,et al.  BLAST+: architecture and applications , 2009, BMC Bioinformatics.

[11]  R. Conrad,et al.  Bacteria rather than Archaea dominate microbial ammonia oxidation in an agricultural soil. , 2009, Environmental microbiology.

[12]  S. Recous,et al.  Shifts between Nitrospira- and Nitrobacter-like nitrite oxidizers underlie the response of soil potential nitrite oxidation to changes in tillage practices. , 2010, Environmental microbiology.

[13]  J. Prosser Soil Nitrifiers and Nitrification , 2011 .

[14]  Andreas Richter,et al.  amoA-based consensus phylogeny of ammonia-oxidizing archaea and deep sequencing of amoA genes from soils of four different geographic regions , 2012, Environmental microbiology.

[15]  D. Myrold,et al.  Dynamics of ammonia-oxidizing archaea and bacteria populations and contributions to soil nitrification potentials , 2012, The ISME Journal.

[16]  Susan Holmes,et al.  phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data , 2013, PloS one.

[17]  Jordan A. Fish,et al.  Ecological Patterns of nifH Genes in Four Terrestrial Climatic Zones Explored with Targeted Metagenomics Using FrameBot, a New Informatics Tool , 2013, mBio.

[18]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[19]  R. Conrad,et al.  Niche differentiation of ammonia oxidizers and nitrite oxidizers in rice paddy soil. , 2013, Environmental microbiology.

[20]  Robert C. Edgar,et al.  UPARSE: highly accurate OTU sequences from microbial amplicon reads , 2013, Nature Methods.

[21]  Jordan A. Fish,et al.  FunGene: the functional gene pipeline and repository , 2013, Front. Microbiol..

[22]  Julien F. Ollivier,et al.  Effects of repeated application of sulfadiazine-contaminated pig manure on the abundance and diversity of ammonia and nitrite oxidizers in the root-rhizosphere complex of pasture plants under field conditions , 2013, Front. Microbio..

[23]  Andreas Richter,et al.  NxrB encoding the beta subunit of nitrite oxidoreductase as functional and phylogenetic marker for nitrite-oxidizing Nitrospira. , 2014, Environmental microbiology.

[24]  James R. Cole,et al.  Ribosomal Database Project: data and tools for high throughput rRNA analysis , 2013, Nucleic Acids Res..

[25]  P. Nielsen,et al.  Complete nitrification by a single microorganism , 2015, Nature.

[26]  M. Wagner,et al.  Complete nitrification by Nitrospira bacteria , 2015, Nature.

[27]  D. Stahl,et al.  Evaluation of revised polymerase chain reaction primers for more inclusive quantification of ammonia-oxidizing archaea and bacteria. , 2015, Environmental microbiology reports.

[28]  P. Christie,et al.  Effects of 44 years of chronic nitrogen fertilization on the soil nitrifying community of permanent grassland , 2015 .

[29]  Jordan A. Fish,et al.  Xander: employing a novel method for efficient gene-targeted metagenomic assembly , 2015, Microbiome.

[30]  M. Habteselassie,et al.  Ammonia-oxidizing bacteria are more responsive than archaea to nitrogen source in an agricultural soil , 2016 .

[31]  Yang Ouyang Agricultural nitrogen management affects microbial communities, enzyme activities, and functional genes for nitrification and nitrogen mineralization , 2016 .

[32]  S. Hart,et al.  Meta-analysis reveals ammonia-oxidizing bacteria respond more strongly to nitrogen addition than ammonia-oxidizing archaea , 2016 .

[33]  M. Wagner,et al.  A New Perspective on Microbes Formerly Known as Nitrite-Oxidizing Bacteria. , 2016, Trends in microbiology.

[34]  M. Wagner,et al.  AmoA-Targeted Polymerase Chain Reaction Primers for the Specific Detection and Quantification of Comammox Nitrospira in the Environment , 2017, bioRxiv.

[35]  J. Stark,et al.  Ammonium availability and temperature control contributions of ammonia oxidizing bacteria and archaea to nitrification in an agricultural soil , 2017 .

[36]  I-Min A. Chen,et al.  IMG/M: integrated genome and metagenome comparative data analysis system , 2016, Nucleic Acids Res..

[37]  AmoA-targeted polymerase chain reaction primers for the specific detection and quantification of comammox Nitrospira in the environment , 2017 .

[38]  K. Konstantinidis,et al.  Year-Round Shotgun Metagenomes Reveal Stable Microbial Communities in Agricultural Soils and Novel Ammonia Oxidizers Responding to Fertilization , 2017, Applied and Environmental Microbiology.

[39]  Q. Huang,et al.  Shifts in Nitrobacter- and Nitrospira-like nitrite-oxidizing bacterial communities under long-term fertilization practices , 2018, Soil Biology and Biochemistry.

[40]  R. Mulvaney Nitrogen-Inorganic Forms , 2018, SSSA Book Series.

[41]  M. Friesen,et al.  Effect of nitrogen fertilization on the abundance of nitrogen cycling genes in agricultural soils: A meta-analysis of field studies , 2018, Soil Biology and Biochemistry.

[42]  James R Cole,et al.  Nonpareil 3: Fast Estimation of Metagenomic Coverage and Sequence Diversity , 2018, mSystems.

[43]  J. Prosser,et al.  The application of high-throughput sequencing technology to analysis of amoA phylogeny and environmental niche specialisation of terrestrial bacterial ammonia-oxidisers , 2019, Environmental Microbiome.

[44]  J. Norton,et al.  Controls and Adaptive Management of Nitrification in Agricultural Soils , 2019, Front. Microbiol..

[45]  J. Norton,et al.  Short-Term Nitrogen Fertilization Affects Microbial Community Composition and Nitrogen Mineralization Functions in an Agricultural Soil , 2019, Applied and Environmental Microbiology.