The impacts of inorganic nitrogen application on mineralization of C-labelled maize and glucose, and on priming effect in saline alkaline soil

Organic matter dynamics and nutrient availability in saline alkaline soil of the former lake Texcoco will determine the success of a planned reforestation program. Uniformly labelled ¹⁴C-maize (MAI-treatment) and glucose (GLU-treatment) with or without 200 mg  kg⁻¹ soil (MAI-N treatment and GLU-N treatment, respectively) were added to soils with electrolytic conductivity (EC) 56 dS m⁻¹ (soil A) and 12 dS m⁻¹ (soil B) to investigate the importance of N availability on decomposition of organic material. Production of CO₂ and and inorganic N dynamics were monitored. The amount of ¹⁴C-glucose mineralized increased 1.8-times in the soil A, but had no effect in the soil B when 200 mg  kg⁻¹ soil was added, while the amount of ¹⁴C-maize mineralized increased 1.7 and 1.3-times when 200  kg⁻¹ soil was added in the soils A and B, respectively. Application of increased priming effect 3.7-times in the MAI-treatment of the soil A and 3.4-times in the GLU-treatment, while in the soil B the increase of priming effect was 4.1-times in the MAI-treatment and 3.7-times in the GLU-treatment. Of the 200 mg  kg⁻¹ added to both soils less than 10 mg NH₃-N kg⁻¹ was volatilized within one day, while 22 and 44 mg  kg⁻¹ soil was fixed on the soil matrix in the soil A and the soil B, respectively. Therefore more than 100 mg −N kg⁻¹ was immobilized into the microbial biomass within the first day. Concentration of nitrite increased sharply in all the treatments of soil A at the onset of the incubation followed by a decrease. A similar pattern was observed in the GLU-N and MAI-N treatments of the soil B, but not in the other treatments. A decrease in concentration of was observed in both soils followed by an increase in the MAI-N and GLU-N treatments of the soil B. It was found that application of had a stimulating effect on the decomposition of maize and glucose, and on the priming effect, while assimilatory reduction of resulted in an increase of in the soil A, and nitrification in the soil B.     

Developing a Technique to Quantify Heterotrophic and Autotrophic Nitrification in Acidic Pasture Soils

Abstract

A series of laboratory incubation experiments were conducted on soils from Maindample and Ruffy in northeast Victoria and from Whittlesea in the Plenty Valley, north of Melbourne, Victoria, Australia, to develop a technique for quantifying both autotrophic and heterotrophic nitrification in acidic pasture soils. The use of a specific inhibitor of the autotrophic ammonium oxidizers (N‐serve) did not completely inhibit autotrophic nitrification in its commonly recommended concentrations (10 and 20 µg g^?1 soil) in these soils. The N‐serve concentration, which completely inhibited autotrophic nitrification, was found to be 60–80 µg g^?1. Varying soil types, pHs, and organic‐matter contents affected the optimum dose of N‐serve required for complete inhibition of autotrophic nitrification. Mixing the inhibitor with the soil after application was also important for immediate inhibition of autotrophic nitrification. Using N‐serve in combination with ¹⁵N‐labeled glycine in the Maindample soil showed that heterotrophic organisms were using the organic route for nitrification, and N‐serve did not affect heterotrophic nitrification. A lag of 12 to 24 h in complete inhibition of autotrophic nitrification by N‐serve may have occurred suggesting nitrification studies using N‐serve should include pre‐incubation of the soils with N‐serve for at least 1 day.     

Microbial activity in pig slurry-amended soils under semiarid conditions

Soil amendment with manures from intensive animal industries is nowadays a common practice that may favorably or adversely affect several soil properties, including soil microbial activity. In this work, the effect of consecutive annual additions of pig slurry (PS) at rates of 30, 60, 90, 120 and 150 m³ ha⁻¹ y⁻¹ over a 4-year period on soil chemical properties and microbial activity was investigated and compared to that of an inorganic fertilization and a control (without amendment). Field plot experiment conducted under a continuous barley monoculture and semiarid conditions were used. Eight months after the fourth yearly PS and mineral fertilizer application (i.e. soon after the fourth barley harvest), surface soil samples (Ap horizon, 0-15 cm depth) from control and amended soils were collected and analysed for pH, electrical conductivity (EC), contents of total organic C, total N, available P and K, microbial biomass C, basal respiration and different enzymatic activities. The control soil had a slightly acidic pH (6.0), a small EC (0.07 dS m⁻¹), adequate levels of total N (1.2 g kg⁻¹) and available K (483 mg kg⁻¹) for barley growth, and small contents of total organic C (13.2 g kg⁻¹) and available P (52 mg kg⁻¹). With respect to the control and mineral fertilized soils, the PS-amended soils had greater pH values (around neutrality or slightly alkaline), electrical conductivities (still low) and contents of available P and K, and slightly larger total N contents. A significant decrease of total organic C was observed in soils amended at high slurry rate (12.3 g kg⁻¹). Compared with the control and mineral treatments, which produced almost similar results, the PS-amended soils were characterized by a higher microbial biomass C content (from 311 to 442 g kg⁻¹), microbial biomass C/total organic C ratio (from 2.3 to 3.6%) and dehydrogenase (from 35 to 173 μg INTF g⁻¹), catalase (from 5 to 24 μmol O₂ g⁻¹ min⁻¹), BAA-protease (from 0.7 to 1.9 μmol  g⁻¹ h⁻¹) and β-glucosidase (from 117 to 269 μmol PNP g⁻¹ h⁻¹) activities, similar basal respirations (from 48 to 77 μg C-CO₂ g⁻¹ d⁻¹) and urease activities (from 1.5 to 2.2 μmol  g⁻¹ h⁻¹), and smaller metabolic quotients (from 6.4 to 7.7 ng C-CO₂ μg⁻¹ biomass C h⁻¹) and phosphatese activities (from 374 to 159 μmol PNP g⁻¹ h⁻¹). For example, statistical analysis of experimental data showed that, with the exception of metabolic quotient and total organic C content, these effects generally increased with increasing cumulative amount of PS. In conclusion, cumulative PS application to soil over time under semiarid conditions may produce not only beneficial effects but also adverse effects on soil properties, such us the partial mineralization of soil organic C through extended microbial oxidation. Thus, PS should not be considered as a mature organic amendment and should be treated appropriately before it is applied to soil, so as to enhance its potential as a soil organic fertilizer.     

Contribution of nitrification and denitrification to N2O emissions from urine patches

Urine deposition by grazing livestock causes an immediate increase in nitrous oxide (N₂O) emissions, but the responsible mechanisms are not well understood. A nitrogen-15 (¹⁵N) labelling study was conducted in an organic grass-clover sward to examine the initial effect of urine on the rates and N₂O loss ratio of nitrification (i.e. moles of N₂O-N produced per moles of nitrate produced) and denitrification (i.e. moles of N₂O produced per moles of N₂O+N₂ produced). The effect of artificial urine (52.9 g N m⁻²) and ammonium solution (52.9 g N m⁻²) was examined in separate experiments at 45% and 35% water-filled pore space (WFPS), respectively, and in each experiment a water control was included. The N₂O loss derived from nitrification or denitrification was determined in the field immediately after application of ¹⁵N-labelled solutions. During the next 24 h, gross nitrification rates were measured in the field, whereas the denitrification rates were measured in soil cores in the laboratory. Compared with the water control, urine application increased the N₂O emission from 3.9 to 42.3 μg N₂O-N m⁻² h⁻¹, whereas application of ammonium increased the emission from 0.9 to 6.1 μg N₂O-N m⁻² h⁻¹. In the urine-affected soil, nitrification and denitrification contributed equally to the N₂O emission, and the increased N₂O loss resulted from a combination of higher rates and higher N₂O loss ratios of the processes. In the present study, an enhanced nitrification rate seemed to be the most important factor explaining the high initial N₂O emission from urine patches deposited on well-aerated soils.     

Emission of N2O, N2 and CO2 from soil fertilized with nitrate: effect of compaction, soil moisture and rewetting

Soil compaction and soil moisture are important factors influencing denitrification and N₂O emission from fertilized soils. We analyzed the combined effects of these factors on the emission of N₂O, N₂ and CO₂ from undisturbed soil cores fertilized with (150 kg N ha⁻¹) in a laboratory experiment. The soil cores were collected from differently compacted areas in a potato field, i.e. the ridges (ρ_D=1.03 g cm⁻³), the interrow area (ρ_D=1.24 g cm⁻³), and the tractor compacted interrow area (ρ_D=1.64 g cm⁻³), and adjusted to constant soil moisture levels between 40 and 98% water-filled pore space (WFPS).High N₂O emissions were a result of denitrification and occurred at a WFPS≥70% in all compaction treatments. N₂ production occurred only at the highest soil moisture level (≥90% WFPS) but it was considerably smaller than the N₂O-N emission in most cases. There was no soil moisture effect on CO₂ emission from the differently compacted soils with the exception of the highest soil moisture level (98% WFPS) of the tractor-compacted soil in which soil respiration was significantly reduced. The maximum N₂O emission rates from all treatments occurred after rewetting of dry soil. This rewetting effect increased with the amount of water added. The results show the importance of increased carbon availability and associated respiratory O₂ consumption induced by soil drying and rewetting for the emissions of N₂O.     

Constraining the conditions conducive to dissimilatory nitrate reduction to ammonium in temperate arable soils

Here we offer the first assessment of conditions conducive to dissimilatory nitrate reduction to ammonium (DNRA) in temperate arable soils, through an examination of the potential for this process to occur in a range of soils of contrasting characteristics. NH₄¹⁵NO₃ (6.2 g N m⁻², 25 atom % excess ¹⁵N) was applied, and recovery of ¹⁵N in the pool taken as indicative of occurrence of DNRA. Up to 5% of applied ¹⁵N was recovered in the pool 2 d after addition of N, glucose (44.6 g C m⁻²) and l-cysteine (7.7 g m⁻², 0.9 g N m⁻², 2.3 g C m⁻²). concentrations were positively correlated with soil pH, ratio, bulk density, sand content and concentration, but negatively correlated with soil C and organic N content. Our results demonstrate the potential for DNRA to contribute to N cycling in temperate arable soils, but its detection and significance is likely to depend on the provision of a low molecular weight C source.     

Initial recovery of soil carbon and nitrogen pools and dynamics following disturbance in jack pine forests: A comparison of wildfire and clearcut harvesting

Forests naturally maintained by stand-replacing wildfires are often managed with clearcut harvesting, yet we know little about how replacing wildfire with clearcutting affects soil processes and properties. We compared the initial recovery of carbon (C) and nitrogen (N) pools and dynamics following disturbance in jack pine (Pinus banksiana) stands in northern Lower Michigan, USA, by sampling soils (Oa+A horizons) from three “treatments”: 3-6-year-old harvest-regenerated stands, 3-6-year-old wildfire-regenerated stands and 40-55-year-old intact, mature stands (n=4 stands per treatment). We measured total C and N; microbial biomass and potentially mineralizable C and N; net nitrification; and gross rates of N mineralization and nitrification. Burned stands exhibited reduced soil N but not C, whereas clearcut and mature stands had similar quantities of soil organic matter. Both disturbance types reduced microbial biomass C compared to mature stands; however, microbial biomass N was reduced in burned stands but not in clearcut stands. The experimental C and N mineralization values were fit to a first-order rate equation to estimate potentially mineralizable pool size (C₀ and N₀) and rate parameters. Values for C₀ in burned and clearcut stands were approximately half that of the mature treatment, with no difference between disturbance types. In contrast, N₀ was lowest in the wildfire stands (170.2 μg N g⁻¹), intermediate in the clearcuts (215.4 μg N g⁻¹) and highest in the mature stands (244.6 μg N g⁻¹). The most pronounced difference between disturbance types was for net nitrification. These data were fit to a sigmoidal growth equation to estimate potential NO₃⁻ accumulation (Nit_max) and kinetic parameters. Values of Nit_max in clearcut soils exceeded that of wildfire and mature soils (149.2 vs. 83.5 vs. 96.5 μg NO₃⁻-N g⁻¹, respectively). Moreover, the clearcut treatment exhibited no lag period for net NO₃⁻ production, whereas the burned and mature treatments exhibited an approximate 8-week lag period before producing appreciable quantities of NO₃⁻. There were no differences between disturbances in gross rates of mineralization or nitrification; rather, lower NO₃⁻ immobilization rates in the clearcut soils, 0.20 μg NO₃⁻ g⁻¹ d⁻¹ compared to 0.65 in the burned soils, explained the difference in net nitrification. Because the mobility of NO₃⁻ and NH₄⁺ differs markedly in soil, our results suggest that differences in nitrification between wildfire and clearcutting could have important consequences for plant nutrition and leaching losses following disturbance.     

Forest floor gross and net nitrogen mineralization in three forest types in Quebec, Canada

The effect of high nitrogen (N) depositions on forest ecosystems is an important concern in North America and may lead to N saturation of forest ecosystems and contribute to soils and surface water acidification. In this study, nitrogen dynamics in the FH layers of a sugar maple (SM), a balsam fir (BF) and a black spruce (BS) forest was characterized using a short term ¹⁵N isotopic pool dilutions approach and mid-term FH material incubation both in situ and in the laboratory. The short term dilutions approach indicated that the mean residence times of and in the FH material of the three sites were low (<1 d). The amount of inorganic nitrogen () recycled annually within the exchangeable forest floor reservoir was between one and two orders of magnitude larger than the annual atmospheric N deposition found at each of the sites. The BS site was clearly distinct than the two other forest types in that net N mineralization was negligible, even in absence of root uptake, suggesting that soil microorganisms were severely N limited. While net nitrification was not observed within the FH material of the BF site, did accumulate in the FH of the SM despite a low pH of 3.72 presumably because of heterotrophic nitrification or as a result of acid-tolerant autotrophic nitrification. The difference in N dynamics between the sites were most probably caused by dominant tree species. Transformation rates of inorganic N were higher in SM, followed by BF and BS stands. Given that the potential to mineralize inorganic N matches with a superimposed N atmospheric deposition gradient in Québec, the sugar maple forest is more likely to be affected by N saturation than coniferous forests.     

A survey of symbiotic nitrogen fixation by white clover grown on metal contaminated soils

There is conflicting evidence about toxic effects of heavy metals in soil on symbiotic nitrogen fixation. This study was set-up to assess the general occurrence of such effects. Soils with metal concentration gradients were sampled from six established field trials, where sewage sludge or metal salts have been applied, or from a transect in a sludge treated soil. Additional contaminated soils were sampled near metal smelters, in floodplains, in sludge amended arable land and in a metalliferous area. Symbiotic nitrogen fixation was measured with ¹⁵N isotope dilution in white clover (Trifolium repens L.) grown in potted soil that was not re-inoculated, and using ryegrass (Lolium perenne L.) as reference crop. The fraction nitrogen in clover derived from fixation (N_dff) varied from 0 to 88% depending on soil. Pronounced metal toxicity on N_dff was only confirmed in a sludge treated soil where nitrogen fixation was halved from the control value at soil total metal concentration of 737 mg Zn kg⁻¹, 428 mg Cu kg⁻¹ and 10 mg Cd kg⁻¹. The N_dff was significantly reduced by increasing metal concentration in soils from two other sites where N_dff was low throughout and where these effects might be attributed to confounding factors. No significant effects of metals on N_dff were identified in all other gradients even up to elevated total metal concentration (e.g. 55 mg Cd kg⁻¹). The variation of N_dff among all soils (n=48), is mainly explained by the number of rhizobia in the soil (log MPN, log (cells g⁻¹ soil)), whereas correlations with total or soil solution metal concentrations were weak (R²<0.25). The is significantly affected by the presence or absence of the host plant at the sampling site. No effects of metals were identified at even at total Zn concentrations of about 2000 mg Zn kg⁻¹, whereas metal toxicity could be identified at lower most probable number (MPN) values. This survey shows that the metal toxicity on symbiotic nitrogen fixation cannot be generalized and that survival of a healthy population of the microsymbiont is probably the critical factor.     

Gross nitrogen transformations in adjacent native and plantation forests of subtropical Australia

The impact of land-use change on soil nitrogen (N) transformations was investigated in adjacent native forest (NF), 53 y-old first rotation (1R) and 5 y-old second rotation (2R) hoop pine (Araucaia cunninghamii) plantations. The ¹⁵N isotope dilution method was used to quantify gross rates of N transformations in aerobic and anaerobic laboratory incubations. Results showed that the land-use change had a significant impact on the soil N transformations. Gross ammonification rates in the aerobic incubation ranged between 0.62 and 1.78 mg N kg⁻¹ d⁻¹, while gross nitrification rates ranged between 2.1 and 6.6 mg N kg⁻¹ d⁻¹. Gross ammonification rates were significantly lower in the NF and the 1R soils than in the 2R soils, however gross nitrification rates were significantly higher in the NF soils than in the plantation soils. The greater rates of gross nitrification found in the NF soil compared to the plantation soils, were related to lower soil C:N ratios (i.e. more labile soil N under NF). Nitrification was found to be the dominant soil N transformation process in the contrasting forest ecosystems. This might be attributed to certain site conditions which may favour the nitrifying community, such as the dry climate and tree species. There was some evidence to suggest that heterotrophic nitrifiers may undertake a significant portion of nitrification.     

Nitrite production by Nitrosomonas europaea and Nitrosospira sp. AV in soils at different solution concentrations of ammonium

Although it remains unclear why NH₃-oxidizing bacteria (AOB) of the genus Nitrosospira dominate soil environments, and why Nitrosomonas spp. are less common, virtually no studies have compared their behavior in soil. In this study, the NH₃ oxidation rates of Nitrosomonas europaea (ATCC 19718) and Nitrosospira sp. AV were compared in three differently textured soils containing a range of extractable contents (2-11 μg soil). Soils were adjusted to pH 7.0-7.4 with CaCO₃ and sterilized with γ-radiation. Cell suspensions of each bacterium were inoculated into the soils to bring them to two-third of water-holding capacity and cell densities ∼2.5×10⁶ g⁻¹ soil. In virtually all cases, rates of production for both N. europaea and Nitrosospira sp. AV were linear over 48 h, and represented between 13 and 75%, respectively, of the maximum rates achieved in soil-free bacterial suspensions. Soil solution concentrations that supported these rates ranged between 0.2 and 1.5 mM. Addition of 21-36 μg soil raised soil solution levels to 1.8-2.5 mM and stimulated production to a greater extent in N. europaea (3.3-6.6-fold) than in Nitrosospira sp. AV (1-2.1-fold). Maximum rates of production were obtained by raising soil solution levels to 3-4 mM with a supplement of ∼80-90 μg soil. K_s values in soil for Nitrosospira sp. AV and N. europaea were estimated as 0.14 and 1.9 mM , respectively, and estimates of V_max were about 3.5-times higher for N. europaea (0.007 pmol h⁻¹ cell⁻¹) than for Nitrosospira sp. AV (0.002 pmol h⁻¹ cell⁻¹). The cell density of N. europaea increased in sterile Steiwer soil independent of supplemental . In the case of treatments receiving supplemental , growth yields of N. europaea calculated from either produced or consumed were similar to those reported in literature (3.5×10⁶-6×10⁶ cells μmol⁻¹). A higher growth yield was measured in the case of zero added (2.7×10⁷ cells μmol⁻¹), indicating that use of organic carbon compounds might have occurred and resulted in some energy sparing. Our results suggest that Nitrosospira spp. with a K_s similar to Nitrosospira sp. AV may have an advantage for survival in soil environments where soil solution levels are less than 1 mM. However, it is apparent that AOB like N. europaea are poised to take advantages of modest increases in extractable that raise soil solution levels to about 2.0-2.5 mM.     

The role of plant residues in pH change of acid soils differing in initial pH

Reports on the effect of plant residues on soil pH have been contradictory. The conflicting accounts have been suggested to result from differences in compositions and types of plant residues and characteristics of soils. This incubation study examined the effect of plant residues differing in concentrations of N (3-49 g kg⁻¹) and of alkalinity (excess cations) (220-1560 mmol kg⁻¹) on pH change of three soils differing in initial pH (3.9-5.1 in 0.01 M CaCl₂). The addition of plant residues at a rate of 15 g kg⁻¹ soil weight increased the pH of all soils by up to 3.4 units and the pH reached the maximum at day 42 after incubation for Wodjil (initial pH 3.87) and Bodallin (pH 4.54) soils and day 14 for Lancelin soil (pH 5.1). The amount of pH buffering was decreased by residue addition in Wodjil soil, increased in Bodallin soil and remained unchanged in Lancelin soil, which closely related to changes of soil pH. Residue addition increased concentration and the increase in concentration generally correlated positively with the concentration of residue N. The concentration increased with time, reached the peak at Days 42-105 for Wodjil soil, Days 14-105 for Bodallin soil and Days 14-42 for Lancelin soil, and then decreased only in Lancelin soil. The concentration of was kept minimal in Wodjil and Bodallin soils. In Lancelin soil, concentrations increased with incubation time from days 14-28. Irrespective of plant residue and incubation time, the amounts of alkalinity produced due to residue addition correlated highly with the sum of the alkalinity added as plant residues (excess cations) and those resulting from mineralization of residue N, with the slope of regression lines decreasing with increase of the initial soil pH. Direct shaking of soil with the residues at the same rate of alkalinity (excess cations) under sterile conditions increased the pH of the Wodjil soil but decreased it in the Lancelin soil. It is suggested that the decarboxylation of organic anions (as indicated by excess cations) of added plant residues and ammonification of the residue N causes soil pH increase whereas nitrification of mineralised residue nitrogen causes soil pH decrease, and that the association/dissociation of organic compounds also plays a role in soil pH change, depending initial pH of the soil. The overall effect on soil pH after addition of plant residues would therefore depend on the extent of each of these processes under given conditions.     

N2O emissions and product ratios of nitrification and denitrification as affected by freezing and thawing

Agricultural soils contribute significantly to atmospheric nitrous oxide (N₂O). A considerable part of the annual N₂O emission may occur during the cold season, possibly supported by high product ratios in denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O-N/(NO₃⁻-N+NO₂⁻-N)) at low temperatures and/or in response to freeze-thaw perturbation. Water-soluble organic materials released from frost-sensitive catch crops and green manure may further increase winter emissions. We conducted short-term laboratory incubations under standardized moisture and oxygen (O₂) conditions, using nitrogen (N) tracers (¹⁵N) to determine process rates and sources of emitted N₂O after freeze-thaw treatment of soil or after addition of freeze-thaw extract from clover. Soil respiration and N₂O production was stimulated by freeze-thaw or addition of plant extract. The N₂O emission response was inversely related to O₂ concentration, indicating denitrification as the quantitatively prevailing process. Denitrification product ratios in the two studied soils (pH 4.5 and 7.0) remained largely unaltered by freeze-thaw or freeze-thaw-released plant material, refuting the hypothesis that high winter emissions are due to frost damage of N₂O reductase activity. Nitrification rates estimated by nitrate (NO₃⁻) pool enrichment were 1.5-1.8 μg NO₃-N g⁻¹ dw soil d⁻¹ in freeze-thaw-treated soil when incubated at O₂ concentrations above 2.3 vol% and one order of magnitude lower at 0.8 vol% O₂. Thus, the experiments captured a situation with severely O₂-limited nitrification. As expected, the O₂ stress at 0.8 vol% resulted in a high nitrification product ratio (0.3 g g⁻¹). Despite this high product ratio, only 4.4% of the measured N₂O accumulation originated from nitrification, reaffirming that denitrification was the main N₂O source at the various tested O₂ concentrations in freeze-thaw-affected soil. N₂O emission response to both freeze-thaw and plant extract addition appeared strongly linked to stimulation of carbon (C) respiration, suggesting that freeze-thaw-induced release of decomposable organic C was the major driving force for N₂O emissions in our soils, both by fuelling denitrifiers and by depleting O₂. The soluble C (applied as plant extract) necessary to induce a CO₂ and N₂O production rate comparable with that of freeze-thaw was 20-30 μg C g⁻¹ soil dw. This is in the range of estimates for over-winter soluble C loss from catch crops and green manure plots reported in the literature. Thus, freeze-thaw-released organic C from plants may play a significant role in freeze-thaw-related N₂O emissions.     

CH4 oxidation and N2O emissions at varied soil water-filled pore spaces and headspace CH4 concentrations

Emission of N₂O and CH₄ oxidation rates were measured from soils of contrasting (30-75%) water-filled pore space (WFPS). Oxidation rates of ¹³C-CH₄ were determined after application of 10 μl ¹³C-CH₄ l⁻¹ (10 at. % excess ¹³C) to soil headspace and comparisons made with estimates from changes in net CH₄ emission in these treatments and under ambient CH₄ where no ¹³C-CH₄ had been applied. We found a significant effect of soil WFPS on ¹³C-CH₄ oxidation rates and evidence for oxidation of 2.2 μg ¹³C-CH₄ d⁻¹ occurring in the 75% WFPS soil, which may have been either aerobic oxidation occurring in aerobic microsites in this soil or anaerobic CH₄ oxidation. The lowest ¹³C-CH₄ oxidation rate was measured in the 30% WFPS soil and was attributed to inhibition of methanotroph activity in this dry soil. However, oxidation was lowest in the wetter soils when estimated from changes in concentration of ¹²⁺¹³C-CH₄. Thus, both methanogenesis and CH₄ oxidation may have been occurring simultaneously in these wet soils, indicating the advantage of using a stable isotope approach to determine oxidation rates. Application of ¹³C-CH₄ at 10 μl ¹³C-CH₄ l⁻¹ resulted in more rapid oxidation than under ambient CH₄ conditions, suggesting CH₄ oxidation in this soil was substrate limited, particularly in the wetter soils. Application of and (80 mg N kg soil⁻¹; 9.9 at.% excess ¹⁵N) to different replicates enabled determination of the respective contributions of nitrification and denitrification to N₂O emissions. The highest N₂O emission (119 μg ¹⁴⁺¹⁵N-N₂O kg soil⁻¹ over 72 h) was measured from the 75% WFPS soil and was mostly produced during denitrification (18.1 μg ¹⁵N-N₂O kg soil⁻¹; 90% of ¹⁵N-N₂O from this treatment). Strong negative correlations between ¹⁴⁺¹⁵N-N₂O emissions, denitrified ¹⁵N-N₂O emissions and ¹³C-CH₄ concentrations (r=−0.93 to −0.95, N₂O; r=−0.87 to −0.95, denitrified ¹⁵N-N₂O; P<0.05) suggest a close relationship between CH₄ oxidation and denitrification in our soil, the nature of which requires further investigation.     

Effects of repeated fertilizer and cattle slurry applications over 38 years on N dynamics in a temperate grassland soil

The effects of repeated synthetic fertilizer or cattle slurry applications at annual rates of 50, 100 or 200 m³ ha⁻¹ yr⁻¹ over a 38 year period were investigated with respect to herbage yield, N uptake and gross soil N dynamics at a permanent grassland site. While synthetic fertilizer had a sustained and constant effect on herbage yield and N uptake, increasing cattle slurry application rates increased the herbage yield and N uptake linearly over the entire observation period. Cattle slurry applications, two and four times the recommended rate (50 m³ ha⁻¹ yr⁻¹, 170 kg N ha⁻¹), increased N uptake by 46 and 78%, respectively after 38 years. To explain the long-term effect, a ¹⁵N tracing study was carried out to identify the potential change in N dynamics under the various treatments. The analysis model evaluated process-specific rates, such as mineralization, from two organic-N pools, as well as nitrification from NH₄⁺ and organic-N oxidation. Total mineralization was similar in all treatments. However, while in an unfertilized control treatment more than 90% of NH₄⁺ production was related to mineralization of recalcitrant organic-N, a shift occurred toward a predominance of mineralization from labile organic-N in the cattle slurry treatments and this proportion increased with the increase in slurry application rate. Furthermore, the oxidation of recalcitrant organic-N shifted from a predominant NH₄⁺ production in the control treatment, toward a predominant NO₃⁻ production (heterotrophic nitrification) in the cattle slurry treatments. The concomitant increase in heterotrophic nitrification and NH₄⁺ oxidation with increasing cattle slurry application rate was mainly responsible for the increase in net NO₃⁻ production rate. Thus the increase in N uptake and herbage yield on the cattle slurry treatments could be related to NO₃⁻ rather than NH₄⁺ production. The ¹⁵N tracing study was successful in revealing process-specific changes in the N cycle in relationship to long-term repeated amendments.     

Effects of decreasing water potential on gross ammonification and nitrification in an acid coniferous forest soil

Changes in the soil water regime, predicted as a consequence of global climate change, might influence the N cycle in temperate forest soils. We investigated the effect of decreasing soil water potentials on gross ammonification and nitrification in different soil horizons of a Norway spruce forest and tested the hypotheses that i) gross rates are more sensitive to desiccation in the Oa and EA horizon as compared to the uppermost Oi/Oe horizon and ii) that gross nitrification is more sensitive than gross ammonification. Soil samples were adjusted by air drying to water potentials from about field capacity to around −1.0 MPa, a range that is often observed under field conditions at our site. Gross rates were measured using the ¹⁵N pool dilution technique. To ensure that the addition of solute label to dry soils and the local rewetting does not affect the results by re-mineralization or preferential consumption of ¹⁵N, we compared different extraction and incubation times.T₀ times ranging from 10 to 300 min and incubation times of 48 h and 72 h did not influence the rates of gross ammonification and nitrification. Even small changes of water potential decreased gross ammonification and nitrification in the O horizon. In the EA horizon, gross nitrification was below detection limit and the response of the generally low rates of gross ammonification to decreasing water potentials was minor. In the Oi/Oe horizon gross ammonification and nitrification decreased from 37.5 to 18.3 mg N kg⁻¹ soil d⁻¹ and from 15.4 to 5.6 mg N kg⁻¹ soil d⁻¹ when the water potential decreased from field capacity to −0.8 MPa. In the Oa horizon gross ammonification decreased from 7.4 to 4.0 mg N kg⁻¹ soil d⁻¹ when the water potential reached −0.6 MPa. At such water potential nitrification almost ceased, while in the Oi/Oe horizon nitrification continued at a rather high level. Hence, only in the Oa horizon nitrification was more sensitive to desiccation than ammonification. Extended drought periods that might result from climate change will cause a reduction in gross N turnover rates in forest soils even at moderate levels of soil desiccation.     

A hot-spot approach applied to nitrification in tropical acid soils

Several authors have reported that nitrification in acid soils may be restricted to microsites having a more favorable pH. The aim of this study was to propose a conceptual model of the functioning of nitrification in hot-spots, and to test it with the experimental data obtained in laboratory conditions using twelve tropical unamended and -amended soils with a wide range of pH (from 4.2 to 6.9). Nitrification was also measured in two selected soils where the pH was adjusted from 3.5 to 6.2. The model characterizes the relationship between the nitrification rates in unamended and -amended soils as a function of pH. It is based upon the assumption that nitrification of the coming from N mineralization occurs in the hot-spot (RN_h), and the nitrification of the added occurs in the hot-spot and also in its adjacent surrounding region (RN_s). The experimental design was chosen to be able to estimate both nitrification rates. Soil acidity limited nitrification more in -amended soils than in unamended ones. From our approach, this is due to less favorable conditions for nitrification in the region surrounding the hot-spot. The effect of self-induced acidity on nitrification was not noticeable neither in unamended nor in -amended treatments. The model described well three observations made in the experiments: (i) the minimum pH for nitrification to occur was lower for RN_h (pH<4.2) than for RN_s (pH<4.7), (ii) the RN_h/RN_s ratio increased with the decrease of pH (from 1.5 at pH 6 to 8.5 at pH 4), and (iii) for a given pH, the RN_h/RN_s ratio increased with the decrease of the initial pH of the soil. Among the soil parameters determined in this study (i.e. exchangeable Al, EDTA-extractable Cu and Zn, total C and N), only pH was related to nitrification. However, for a given pH, nitrification varied 3-fold among soils, depending upon their initial pH. This suggests that soil pH as determined on bulk soil is not suitable to predict nitrification in each individual soil, because it is not representative of the acidity level within the hot-spot.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.     

A mass-balance approach to measuring microbial uptake and pools of phosphorus in nutrient-amended soils

Soil microbial biomass P is usually determined through fumigation-extraction (FE), in which partially extractable P from lysed biomass is converted to biomass P using a conversion factor (K_p). Estimation of K_p has been usually based on cultured microorganisms, which may not adequately represent the soil microbial community in either nutrient-poor or in altered carbon and nutrient conditions following fertilisation. We report an alternative approach in which changes in microbial P storage are determined as the residual in a mass balance of extractable P before and after incubation. This approach was applied in three low-fertility sandy soils of southwestern Australia, to determine microbial P immobilisation during 5-day incubations in response to the amendment by 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net P immobilisation during the amended incubations determined to be 18.1, 14.1 and 16.3 μg P g⁻¹ in the three soils, accounting for 70.6-90.5% of P added through amendment. Such estimates do not rely on fumigation and K_p values, but for comparison with the FE method we estimated ‘nominal’ K_p values to be 0.20-0.31 for the soils under the amended conditions. Our results showed that microbial P immobilisation was a dominant process regulating P concentration in soil water following the CNP amendment. The mass-balance approach provides information not only about changes in the microbial P compartment, but also about other major P-pools and their fluxes in regulating soil-water P concentrations under substrate- and nutrient-amended conditions.