Grassland Management Regimens Reduce Small-Scale Heterogeneity and Species Diversity of β-Proteobacterial Ammonia Oxidizer Populations

ABSTRACT The impact of soil management practices on ammonia oxidizer diversity and spatial heterogeneity was determined in improved (addition of N fertilizer), unimproved (no additions), and semi-improved (intermediate management) grassland pastures at the Sourhope Research Station in Scotland. Ammonia oxidizer diversity within each grassland soil was assessed by PCR amplification of microbial community DNA with both ammonia oxidizer-specific, 16S rRNA gene (rDNA) and functional, amoA, gene primers. PCR products were analysed by denaturing gradient gel electrophoresis, phylogenetic analysis of partial 16S rDNA and amoA sequences, and hybridization with ammonia oxidizer-specific oligonucleotide probes. Ammonia oxidizer populations in unimproved soils were more diverse than those in improved soils and were dominated by organisms representing Nitrosospira clusters 1 and 3 and Nitrosomonas cluster 7 (closely related phylogenetically to Nitrosomonas europaea). Improved soils were only dominated by Nitrosospira cluster 3 and Nitrosomonas cluster 7. These differences were also reflected in functional gene (amoA) diversity, with amoA gene sequences of both Nitrosomonas and Nitrosospira species detected. Replicate 0.5-g samples of unimproved soil demonstrated significant spatial heterogeneity in 16S rDNA-defined ammonia oxidizer clusters, which was reflected in heterogeneity in ammonium concentration and pH. Heterogeneity in soil characteristics and ammonia oxidizer diversity were lower in improved soils. The results therefore demonstrate significant effects of soil management on diversity and heterogeneity of ammonia oxidizer populations that are related to similar changes in relevant soil characteristics.

[1]  J. Fuchs,et al.  Sequence of the gene coding for ammonia monooxygenase in Nitrosomonas europaea , 1993, Journal of bacteriology.

[2]  M. Lodhi Additional Evidence of Inhibition of Nitrifiers and Possible Cycling of Inhibitors Produced by Selected Plants in a Climax Community , 1982 .

[3]  G. Kowalchuk,et al.  Analysis of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in coastal sand dunes by denaturing gradient gel electrophoresis and sequencing of PCR-amplified 16S ribosomal DNA fragments , 1997, Applied and environmental microbiology.

[4]  G. G. Pohlman Soil Science Society of America , 1940 .

[5]  G. Kowalchuk,et al.  Comparative Diversity of Ammonia Oxidizer 16S rRNA Gene Sequences in Native, Tilled, and Successional Soils , 1999, Applied and Environmental Microbiology.

[6]  K. Killham,et al.  Nitrification in coniferous forest soils , 1990, Plant and Soil.

[7]  L. Bakken,et al.  Autotrophic ammonium-oxidising bacteria in Swedish mor humus , 1999 .

[8]  D Penny,et al.  Evolution of chlorophyll and bacteriochlorophyll: the problem of invariant sites in sequence analysis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M. Davey,et al.  The nodulation of micro-propagated plants of Parasponia andersonii by tropical legume rhizobia , 1995 .

[10]  W. Boer,et al.  Ureolytic nitrification at low pH by Nitrosospira spec. , 1989, Archives of Microbiology.

[11]  J. Prosser,et al.  Molecular analysis of enrichment cultures of marine ammonia oxidisers. , 1994, FEMS microbiology letters.

[12]  R. Sockett,et al.  Use of Molecular and Isotopic Techniques To Monitor the Response of Autotrophic Ammonia-Oxidizing Populations of the β Subdivision of the Class Proteobacteria in Arable Soils to Nitrogen Fertilizer , 1999, Applied and Environmental Microbiology.

[13]  P. Sale,et al.  The influence of the soil matrix on nitrogen mineralisation and nitrification. I. Spatial variation and a hierarchy of soil properties , 1998 .

[14]  D. Stahl,et al.  Evolutionary relationships among ammonia- and nitrite-oxidizing bacteria , 1994, Journal of bacteriology.

[15]  M.,et al.  AMMONIA OXIDATION AT LOW pH BY ATTACHED POPULATIONS OF NITRIFYING BACTERIA , 1993 .

[16]  T. Martin Embley,et al.  Analysis of β-Subgroup Proteobacterial Ammonia Oxidizer Populations in Soil by Denaturing Gradient Gel Electrophoresis Analysis and Hierarchical Phylogenetic Probing , 1998, Applied and Environmental Microbiology.

[17]  A. Uitterlinden,et al.  Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA , 1993, Applied and environmental microbiology.

[18]  M. Klotz,et al.  Multiple copies of ammonia monooxygenase (amo) operons have evolved under biased AT/GC mutational pressure in ammonia-oxidizing autotrophic bacteria. , 1998, FEMS microbiology letters.

[19]  J. Prosser,et al.  Phylogenetic Differences between Particle-Associated and Planktonic Ammonia-Oxidizing Bacteria of the β Subdivision of the Class Proteobacteria in the Northwestern Mediterranean Sea , 1999, Applied and Environmental Microbiology.

[20]  M. P. Cummings PHYLIP (Phylogeny Inference Package) , 2004 .

[21]  D. Horner,et al.  Iron hydrogenases and the evolution of anaerobic eukaryotes. , 2000, Molecular biology and evolution.

[22]  P. Martikainen Numbers of autotrophic nitrifiers and nitrification in fertilized forest soil , 1985 .

[23]  H. Koops,et al.  Description of a new species of Nitrosococcus , 1990, Archives of Microbiology.

[24]  Suiying Huang,et al.  How Stable Is Stable? Function versus Community Composition , 1999, Applied and Environmental Microbiology.

[25]  Robert S. Burlage,et al.  Techniques in microbial ecology , 1998 .

[26]  J. Lake,et al.  Reconstructing evolutionary trees from DNA and protein sequences: paralinear distances. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[27]  A. Page Methods of soil analysis. Part 2. Chemical and microbiological properties. , 1982 .

[28]  J. Prosser,et al.  Molecular diversity of soil and marine 16S rRNA gene sequences related to beta-subgroup ammonia-oxidizing bacteria , 1996, Applied and environmental microbiology.

[29]  W. Holmes,et al.  Grass: Its Production and Utilization , 1983 .

[30]  Michael P. Cummings,et al.  PAUP* [Phylogenetic Analysis Using Parsimony (and Other Methods)] , 2004 .

[31]  E. L. Rice,et al.  Inhibition of nitrification by climax ecosystems , 1972 .

[32]  S. Macnaughton,et al.  Effect of Toxic Metals on Indigenous Soil β-Subgroup Proteobacterium Ammonia Oxidizer Community Structure and Protection against Toxicity by Inoculated Metal-Resistant Bacteria , 1999, Applied and Environmental Microbiology.

[33]  J. Prosser,et al.  Molecular Analysis of Bacterial Community Structure and Diversity in Unimproved and Improved Upland Grass Pastures , 1999, Applied and Environmental Microbiology.

[34]  J. Tiedje,et al.  Characterization of the Dominant and Rare Members of a Young Hawaiian Soil Bacterial Community with Small-Subunit Ribosomal DNA Amplified from DNA Fractionated on the Basis of Its Guanine and Cytosine Composition , 1998, Applied and Environmental Microbiology.

[35]  G. Muyzer DGGE/TGGE a method for identifying genes from natural ecosystems. , 1999, Current opinion in microbiology.

[36]  W. Doolittle,et al.  Microsporidia are related to Fungi: evidence from the largest subunit of RNA polymerase II and other proteins. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Wagner,et al.  Phylogenetic probes for analyzing abundance and spatial organization of nitrifying bacteria , 1996, Applied and environmental microbiology.

[38]  Katherine L. Gross,et al.  Effects of Agronomic Treatments on Structure and Function of Ammonia-Oxidizing Communities , 2000, Applied and Environmental Microbiology.

[39]  M. Veenhuis,et al.  Nitrification at Low pH by Aggregated Chemolithotrophic Bacteria , 1991, Applied and environmental microbiology.

[40]  Tadao Ando,et al.  Plant Nutrition for Sustainable Food Production and Environment , 2016, Developments in Plant and Soil Sciences.

[41]  K. Yokoyama,et al.  Detection of amo gene of chemolithotrophic ammonia-oxidizers in soil by nested PCR , 1997 .

[42]  R. Weller,et al.  Succession and convergence of biofilm communities in fixed-film reactors treating aromatic hydrocarbons in groundwater , 1997, Applied and environmental microbiology.

[43]  L. Bakken,et al.  Phylogenetic Analysis of Seven New Isolates of Ammonia-Oxidizing Bacteria Based on 16S rRNA Gene Sequences , 1995 .

[44]  J R Saunders,et al.  The phylogeny of autotrophic ammonia-oxidizing bacteria as determined by analysis of 16S ribosomal RNA gene sequences. , 1993, Journal of general microbiology.

[45]  B H Olson,et al.  Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction , 1992, Applied and environmental microbiology.

[46]  G. Kowalchuk,et al.  Nitrogen Cycling and Community Structure of Proteobacterial β-Subgroup Ammonia-Oxidizing Bacteria within Polluted Marine Fish Farm Sediments , 1999, Applied and Environmental Microbiology.

[47]  J. Prosser Autotrophic nitrification in bacteria. , 1989, Advances in microbial physiology.

[48]  J. Saunders,et al.  Amplification of 16S ribosomal RNA genes of autotrophic ammonia-oxidizing bacteria demonstrates the ubiquity of nitrosospiras in the environment. , 1995, Microbiology.

[49]  J. S. Rodwell,et al.  British Plant Communities: British Plant Communities , 2000 .

[50]  K. Wickramasinghe,et al.  Nitrification and autotrophic nitrifying bacteria in acid tea soils , 1979 .