Evidence for Fungal Dominance of Denitrification and Codenitrification in a Grassland Soil

Fungi are capable of nitrification and denitrification and often dominate the microbial biomass of temperate grassland soils. We determined the contributions of bacteria and fungi to N 2 O and N 2 production in a grassland soil from Northern Ireland by combining the substrate-induced respiration inhibition method and the 15 N gas-flux method. Streptomycin (C 21 H 39 N 7 O 12 ) was used as the bacterial inhibitor and cycloheximide (C 15 H 23 NO 4 ) as the fungal inhibitor. By labeling the NH 4 and NO 3 pools, we tested the hypothesis that fungi produce N 2 O and N 2 solely by the reduction of NO 3 . Cycloheximide decreased the flux of N 2 O by 89% and streptomycin decreased the flux by 23%, indicating that fungi were responsible for most of the N 2 O production. All of the N 2 O was derived from NO 3 reduction. Labeled N 2 was only detected in control and streptomycin treatments. The distribution of the 15 N atoms in the labeled N 2 indicated that the source of the labeling was predominantly the NO 3 pool, but that the process of formation was not dominated by denitrification. Codenitrification, where a 15 N atom from labeled nitrogen dioxide (NO 2 ) combines with a N atom from a natural abundance source, was proposed as the process forming labeled N 2 . About 92% of the labeled N 2 was estimated to be due to codenitrification and 8% due to denitrification. The flux of N 2 O was always greater than the flux of N 2 , the mole fraction of N 2 O averaging 0.7. Fungal denitrification could be of ecological significance because N 2 O is the dominant gaseous end product.

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

[2]  K. Domsch,et al.  Quantification of bacterial and fungal contributions to soil respiration , 1973, Archiv für Mikrobiologie.

[3]  T. Clough,et al.  Transformations of inorganic-N in soil leachate under differing storage conditions , 2001 .

[4]  R. Stevens,et al.  Cattle Slurry Affects Nitrous Oxide and Dinitrogen Emissions from Fertilizer Nitrate , 2001 .

[5]  R. Stevens,et al.  Lowering the detection limit for dinitrogen using the enrichment of nitrous oxide , 2001 .

[6]  D. Engelbrecht,et al.  Soil layer-specific variability in net nitrification and denitrification in an acid coniferous forest , 2000, Biology and Fertility of Soils.

[7]  M. Norman,et al.  The utility of ergosterol as a bioindicator of fungi in temperate soils , 2000 .

[8]  Zhemin Zhou,et al.  Oxygen requirement for denitrification by the fungus Fusarium oxysporum , 2000, Archives of Microbiology.

[9]  P. Brookes,et al.  Comparison of substrate induced respiration, selective inhibition and biovolume measurements of microbial biomass and its community structure in unamended, ryegrass-amended, fumigated and pesticide-treated soils , 1999 .

[10]  P. Brookes,et al.  An evaluation of the substrate-induced respiration method , 1999 .

[11]  K. Paustian,et al.  Bacterial and fungal abundance and biomass in conventional and no-tillage agroecosystems along two climatic gradients , 1999 .

[12]  M. Hamamoto,et al.  Denitrification by yeasts and occurrence of cytochrome P450nor in Trichosporon cutaneum. , 1998, FEMS microbiology letters.

[13]  S. Castaldi,et al.  Effect of cycloheximide on N2O and NO3– production in a forest and an agricultural soil , 1998, Biology and Fertility of Soils.

[14]  R. Stevens,et al.  Measuring the mole fraction and source of nitrous oxide in the field , 1998 .

[15]  W. Zumft Cell biology and molecular basis of denitrification. , 1997, Microbiology and molecular biology reviews : MMBR.

[16]  Apportioning nitrous oxide fluxes between nitrification and denitrification using gas-phase mass spectrometry , 1997 .

[17]  R. Stevens,et al.  Determining nitrogen-15 in ammonium by producing nitrous oxide , 1997 .

[18]  G. Marzluf,et al.  Genetic regulation of nitrogen metabolism in the fungi , 1997, Microbiology and molecular biology reviews : MMBR.

[19]  H. Shoun,et al.  Denitrification, a Novel Type of Respiratory Metabolism in Fungal Mitochondrion* , 1996, The Journal of Biological Chemistry.

[20]  Cinthia K. Johnson,et al.  Measuring bacterial and fungal substrate-induced respiration in dry soils , 1996 .

[21]  R. Stevens,et al.  Nitrite transformations during soil extraction with potassium chloride , 1995 .

[22]  H. Shoun,et al.  Denitrification by the fungus Cylindrocarpon tonkinense: anaerobic cell growth and two isozyme forms of cytochrome P-450nor , 1995, Applied and environmental microbiology.

[23]  R. Stevens,et al.  Determining Nitrogen‐15 in Nitrite or Nitrate by Producing Nitrous Oxide , 1994 .

[24]  D. S. Schimel,et al.  7 – Nitrification and Denitrification , 1993 .

[25]  H. Uchiyama,et al.  Denitrification by fungi. , 1992, FEMS microbiology letters.

[26]  K. Hatano,et al.  Co-denitrification by the denitrifying system of the fungus Fusarium oxysporum , 1992 .

[27]  J. Arah New Formulae for Mass Spectrometric Analysis of Nitrous Oxide and Dinitrogen Emissions , 1992 .

[28]  Richard T. Lamar,et al.  Evaluation of methods to extract ergosterol for quantitation of soil fungal biomass , 1992 .

[29]  J. Harris,et al.  Habitat Classification and Soil Restoration Assessment Using Analysis of Soil Microbiological and Physico-chemical Characteristics , 1992 .

[30]  H. Shoun,et al.  Denitrification by the fungus Fusarium oxysporum and involvement of cytochrome P-450 in the respiratory nitrite reduction. , 1991, The Journal of biological chemistry.

[31]  D. Myrold Measuring Denitrification in Soils Using 15N Techniques , 1990 .

[32]  William L. Hargrove,et al.  A substrate-induced respiration (SIR) method for measurement of fungal and bacterial biomass on plant residues , 1990 .

[33]  E. Paul,et al.  Soil microbiology and biochemistry. , 1998 .

[34]  P. Baveye,et al.  Evaluation of Nitrogen‐15 Tracer Techniques for Direct Measurement of Denitrification in Soil: I. Theory , 1988 .

[35]  W. D. Grant,et al.  Use of ergosterol, diaminopimelic acid and glucosamine contents of soils to monitor changes in microbial populations , 1987 .

[36]  W. Grant,et al.  Measurement of ergosterol, diaminopimelic acid and glucosamine in soil: evaluation as indicators of microbial biomass , 1986 .

[37]  S. J. Prosser,et al.  Automated Determination of Nitrogen-15-Labeled Dinitrogen and Nitrous Oxide by Mass Spectrometry , 1986 .

[38]  D. Focht,et al.  15N Kinetic Analysis of N2O Production by Nitrosomonas europaea: an Examination of Nitrifier Denitrification , 1985, Applied and environmental microbiology.

[39]  M. Firestone,et al.  Identification of Heterotrophic Nitrification in a Sierran Forest Soil , 1984, Applied and environmental microbiology.

[40]  R. Mulvaney Determination of 15N‐Labeled Dinitrogen and Nitrous Oxide with Triple‐Collector Mass Spectrometers , 1984 .

[41]  D. A. Klein,et al.  Decomposition of Microbial Cell Components in a Semi-Arid Grassland Soil , 1979, Applied and environmental microbiology.

[42]  K. Domsch,et al.  Measurement of bacterial and fungal contributions to respiration of selected agricultural and forest soils. , 1975, Canadian journal of microbiology.

[43]  J. Bollag,et al.  Nitrous oxide release by soil fungi , 1972 .

[44]  Heterotrophic nitrification in soils— a preliminary investigation , 1970 .

[45]  D. Kirkham,et al.  Equations for Following Nutrient Transformations in Soil, Utilizing Tracer Data: II. 1 , 1954 .

[46]  E. Kornfeld,et al.  The Structure of Actidione, An Antibiotic From Streptomyces griseus. , 1948, Science.

[47]  D. Morris Quantitative Determination of Carbohydrates With Dreywood's Anthrone Reagent. , 1948, Science.