A N tracing model to analyse N transformations in old grassland soil

A new ¹⁵N tracing model was developed to analyse nitrogen (N) transformations in old grassland soil. There was a need to develop a new model because existing models such as FLUAZ were not able to simulate the observed N dynamics. The new features of the model are: (a) simulation of heterotrophic nitrification, (b) simulation of dissimilatory nitrate (NO₃⁻) reduction to ammonium (NH₄⁺) (DNRA), (c) release of adsorbed or stored fertiliser N into the available mineral N pools and (d) immobilisation of NH₄⁺ and NO₃⁻ into two separate organic N pools with different re-mineralisation characteristics. The tracing model contains six N pools and nine simultaneous N transformations either at zero- or first-order kinetics. The model is set up in the modelling software ModelMaker which contains non-linear optimisation routines based on the Marquardt-Levenberg algorithm. The model is able to simulate data obtained from triple labelling studies where either the NH₄⁺, the NO₃⁻ or both pools were labelled with ¹⁵N. The flexible modelling environment allows the user to develop the model further.     

Improving the Accuracy of Diffusion for Inorganic 15N Analyses of Soil Extracts and Water

Sequential diffusion techniques used to speciate inorganic nitrogen-15 (¹⁵N) during soil or water analysis are complicated by incomplete recovery of ammonium (NH₄⁺)-N, introducing error in the subsequent determination of nitrate (NO₃^–)-N. Based on studies to evaluate different strategies for minimizing cross-contamination error in Mason-jar diffusions, a simple cleaning technique was developed that involves an additional 6-h diffusion using 0.6 M boric acid (H₃BO₃) at room temperature following the recovery of NH₄⁺-N. This technique was 60–87% effective for reducing cross-contamination of unlabeled NO₃^–-N by labeled NH₄⁺-N and became more effective for controlling analytical error with decreasing sample volumes, lower NH₄⁺-N enrichment, and larger quantities of NO₃^–-N. When used with the cleaning technique described, sequential diffusions were far superior for ¹⁵N analysis of NO₃^–-N, as compared to the nonsequential approach that involves an isotope dilution calculation after separate diffusions to determine NH₄⁺-N and total mineral N.     

Comparison of NH4 pool dilution techniques to measure gross N fluxes in a coarse textured soil

We compared gross N fluxes by ¹⁵N pool dilution in a coarse-textured agricultural soil when ¹⁵N was applied to the soil NH₄⁺ pool by either: (i) mixing a ¹⁵NH₄NO₃ solution into disturbed soil or (ii) injection of ¹⁵NH₃ gas into intact soil cores. The two techniques produced similar results for gross N mineralization rates indicating that NH₄⁺ production in soil was not altered by soil disturbance, method of application (gas vs. solution), or amount of N applied. This was not the case for immobilization rates, which were twofold higher when ¹⁵N label was applied to the soil NH₄⁺ pool with the mixing technique compared to the injection technique. This was attributed to the fact that more NH₄⁺ was applied with the mixing technique. Estimates of gross nitrification were accompanied by large error terms meaning differences between ¹⁵N labeling methods could not be accurately assessed for this process rate.     

Short‐Term Recovery of Ammonium‐15Nitrogen Applied to a Temperate Forest Inceptisol and Ultisol in East Tennessee,USA

Abstract

The short‐term fate and retention of ammonium (NH₄)‐¹⁵nitrogen (N) applied to two types of forest soils in east Tennessee was investigated. Four ridgetop forests, predominantly oak (Quercus spp.), were studied. Five applications of NH₄‐¹⁵N tracer were made to the forest floor at 2‐ to 4‐week intervals over a 14‐week period in 2004. Nitrogen‐15 recovery in the forest floor, fine roots (<2 mm), and the mineral soil (0–20 cm) was calculated at 6, 21, and 42 weeks after the last application. Most of the ¹⁵N was retained in the forest floor and the mineral soil, with only small amounts (≤2%) found in roots from both soil layers. Recovery of NH₄‐¹⁵N was greater in Inceptisols, which had a wider carbon (C)‐to‐N ratio than Ultisols. For both soil types, higher NH₄‐¹⁵N recoveries and long retention times (half‐lives>100 weeks) indicated the forest floor is an effective filter for atmospheric N inputs.     

Effect of elevated CO2 on soil N dynamics in a temperate grassland soil

The response of terrestrial ecosystems to elevated atmospheric CO₂ is related to the availability of other nutrients and in particular to nitrogen (N). Here we present results on soil N transformation dynamics from a N-limited temperate grassland that had been under Free Air CO₂ Enrichment (FACE) for six years. A ¹⁵N labelling laboratory study (i.e. in absence of plant N uptake) was carried out to identify the effect of elevated CO₂ on gross soil N transformations. The simultaneous gross N transformation rates in the soil were analyzed with a ¹⁵N tracing model which considered mineralization of two soil organic matter (SOM) pools, included nitrification from NH₄⁺ and from organic-N to NO₃⁻ and analysed the rate of dissimilatory NO₃⁻ reduction to NH₄⁺ (DNRA). Results indicate that the mineralization of labile organic-N became more important under elevated CO₂. At the same time the gross rate of NH₄⁺ immobilization increased by 20%, while NH₄⁺ oxidation to NO₃⁻ was reduced by 25% under elevated CO₂. The NO₃⁻ dynamics under elevated CO₂ were characterized by a 52% increase in NO₃⁻ immobilization and a 141% increase in the DNRA rate, while NO₃⁻ production via heterotrophic nitrification was reduced to almost zero. The increased turnover of the NH₄⁺ pool, combined with the increased DNRA rate provided an indication that the available N in the grassland soil may gradually shift towards NH₄⁺ under elevated CO₂. The advantage of such a shift is that NH₄⁺ is less prone to N losses, which may increase the N retention and N use efficiency in the grassland ecosystem under elevated CO₂.     

Mobilization of labelled N in soybean leaves during early podfill

Soybeans accumulate N in vegetative tissues up to pod initiation after which total vegetative N may remain constant during early phases of pod development. Eventually much of the vegetative N is mobilized to the pods. The mobilization of N from vegetative tissue to pods during the first few days of pod development is poorly understood but is important to an overall understanding of soybean N nutrition. The vegetative tissues of field grown soybeans were labelled with ¹⁵N and sampled weekly during the reproductive phase of plant growth. Three foliar applications of (¹⁵NH₄)₂SO₄ were made prior to pod initiation at a combined rate of 3.3 kg N/ha. To immobilize soil N and to increase soybean dependance on N₂‐fixation, sawdust was applied at a rate of 52 t ha^‐1 . Irrigation was required almost weekly because of a shallow soil profile and below normal summer precipitation. Mobilization of vegetative N began immediately upon pod initiation and continued at a linear rate through pod development. It appeared that N₂‐fixation was able to provide approximately half of the N in pods during early podfill. Nitrogen content of vegetative tissue declined as soon as pods began developing.     

Interference by amino acids during the determination of N ammonium in soil

In the study of terrestrial N cycling, NH₄⁺ concentration and ¹⁵N enrichment are routinely determined by colorimetric continuous flow analysis and microdiffusion methods. Amino acids can interfere in these determinations; consequently the aim of the present study was to evaluate the significance of the interference. Glycine and glutamine are key amino acids in soil and were therefore used as ‘models’. Both glycine and glutamine interfered during continuous flow analysis, whereas interference during microdiffusion was of little importance. The effects of interference can be significant, e.g. estimates of gross mineralisation rate were reduced up to 33%, where we allowed for amino acid interference during determination of NH₄⁺ concentration. The potential influence of amino acid interference emphasises that development of continuous flow analysis to increase NH₄⁺ specificity is needed.     

Evidence for acclimation of N cycling to episodic N inputs in anthropogenically-affected intertidal salt marsh sediments

Nitrate removal was compared in anthropogenically-impacted and unimpacted salt marsh sediments in microcosms using the acetylene block technique and ¹⁵N natural abundance measurements. Potential denitrification rates were greater at the impacted site than the unimpacted site at all added NO₃⁻ concentrations (233, 467, or 700 μg N g dw⁻¹). Although the change in concentration of NH₄⁺ over time was small (69-104 μg N g dw⁻¹), the δ¹⁵N of accumulated NH₄⁺ increased significantly (0.26-13.22‰), and was more enriched for all NO₃⁻ treatments in the impacted sediments than the unimpacted site. The impacted site may be acclimated to episodic N inputs, and based on concentrations and ¹⁵N natural abundance had greater denitrification and N cycling than the unimpacted site.     

Development of a Soil N Test for Fertilizer Requirements for Corn Production in Quebec

Corn requires high nitrogen (N) fertilizer use, but no soil N test for fertilizer N requirement is yet available in Quebec. Objectives of this research were (1) to determine the effects of soil nitrate (NO₃ ^?)-N, soil ammonium (NH₄ ⁺)-N, and N fertilizer rates on corn yields and (2) to determine soil sampling times and depths most highly correlated with yields and fertilizer N response under Quebec conditions. Soil samples were taken from 0- to 30-cm and 30- to 60-cm depths at seeding and postseeding (when corn height reached 20 cm) to determine soil NH₄ ⁺ and NO₃ ^? in 44 continuous corn sites fertilized with four rates of N in two replications using a quick test (N-Trak) and a laboratory method. The N-Trak method overestimated soil NO₃ ^?-N in comparison with the laboratory method. Greater coefficients of determination were observed for soil NO₃ ^?-N analyses at postseeding compared with seeding.     

Short-term competition between crop plants and soil microbes for inorganic N fertilizer

Agricultural systems that receive high amounts of inorganic nitrogen (N) fertilizer in the form of either ammonium (NH₄⁺), nitrate (NO₃⁻) or a combination thereof are expected to differ in soil N transformation rates and fates of NH₄⁺ and NO₃⁻. Using ¹⁵N tracer techniques this study examines how crop plants and soil microbes vary in their ability to take up and compete for fertilizer N on a short time scale (hours to days). Single plants of barley (Hordeum vulgare L. cv. Morex) were grown on two agricultural soils in microcosms which received either NH₄⁺, NO₃⁻ or NH₄NO₃. Within each fertilizer treatment traces of ¹⁵NH₄⁺ and ¹⁵NO₃⁻ were added separately. During 8 days of fertilization the fate of fertilizer ¹⁵N into plants, microbial biomass and inorganic soil N pools as well as changes in gross N transformation rates were investigated. One week after fertilization 45-80% of initially applied ¹⁵N was recovered in crop plants compared to only 1-10% in soil microbes, proving that plants were the strongest competitors for fertilizer N. In terms of N uptake soil microbes out-competed plants only during the first 4 h of N application independent of soil and fertilizer N form. Within one day microbial N uptake declined substantially, probably due to carbon limitation. In both soils, plants and soil microbes took up more NO₃⁻ than NH₄⁺ independent of initially applied N form. Surprisingly, no inhibitory effect of NH₄⁺ on the uptake and assimilation of nitrate in both, plants and microbes, was observed, probably because fast nitrification rates led to a swift depletion of the ammonium pool. Compared to plant and microbial NH₄⁺ uptake rates, gross nitrification rates were 3-75-fold higher, indicating that nitrifiers were the strongest competitors for NH₄⁺ in both soils. The rapid conversion of NH₄⁺ to NO₃⁻ and preferential use of NO₃⁻ by soil microbes suggest that in agricultural systems with high inorganic N fertilizer inputs the soil microbial community could adapt to high concentrations of NO₃⁻ and shift towards enhanced reliance on NO₃⁻ for their N supply.     

Evidence of carbon stimulated N transformations in grassland soil after slurry application

High nitrification rates which convert ammonium (NH₄⁺) to the mobile ions NO₂⁻ and NO₃⁻ are of high ecological significance because they increase the potential for N losses via leaching and denitrification. Nitrification can be performed by chemoautotrophic or heterotrophic organisms and heterotrophic nitrifiers can oxidise either mineral (NH₄⁺) or organic N. Selective nitrification inhibitors and ¹⁵N tracer studies have been used in an attempt to separate heterotrophic and autotrophic nitrification. In a laboratory study we determined the effect of cattle slurry on the oxidation of mineral NH₄⁺-N and organic-N by labelling the NH₄⁺ or NO₃⁻ pools separately or both together with ¹⁵N. The size and enrichment of the mineral N pools were determined at intervals. To calculate gross N transformation rates a ¹⁵N tracing model was developed. This model consists of the three N-pools NH₄⁺, NO₃⁻ and organic N. Sub-models for decomposition of degradable carbon in the soil and the slurry were added to the model and linked to the N transformation rates. The model was set up in the software ModelMaker which contains non-linear optimization routines to determine model parameters. The application of cattle slurry increased the rate of nitrifcation by a factor of 20 compared with the control. The size and enrichment of the mineral N pools provided evidence that nitrification was due to the conversion of NH₄⁺ to NO₃⁻ and not the conversion of organic N to NO₃⁻. There was evidence that slurry-enhanced oxidation of NH₄⁺ to NO₃⁻ was due to a combination of autotrophic and heterotrophic transformations. Slurry application increased the mineralisation rate by approximately a factor of two compared with the control and the rate of immobilisation of NH₄⁺ by approximately a factor of three.     

Amino acid interference with ammonium determination in soil extracts using the automated indophenol method

Abstract

The formation of a colored indophenol complex is commonly used as a quantitative measure of the ammonium content of soil extracts. The potential interference with ammonium determination from co‐extracted amino acids was examined. The extent of color development was examined for 22 amino acids by subjecting pure solutions to ammonium determination by both the indophenol method and steam distillation. Apparent detection of amino acid as ammonium ranged from 0 to 94 % of total nitrogen for the indophenol procedure, whereas steam distillation resulted in little apparent ammonium recovery. With the exception of threonine, the extent of color development was inversely related to amino acid molecular weight. The range in recoveries for the indophenol procedure suggests both size and composition of the co‐extracted amino acid pool is important in determining the extent of interference.

Significantly (p=0.001) greater estimates, averaging 0.4 μg mL^‐1, were found in indophenol estimates of mineral‐N content of moist, fresh soil samples. Air drying, oven drying or chloroform fumigation significantly increased the difference (0.3 ‐ 0.7 μg mL^‐1) in estimates of ammonium content. At 10: 1 extract: soil ratios this could cause ? to be overestimated by 3–7 μg g^‐1soil. The increased interference was attributed to a release of amino acid as a result of pretreatment. The difference between distillation and indophenol estimates of ammonium content of 0.5 M K₂SO₄was found to be dependent upon ammonium content. The use of procedures employing a distillation step (manual or automated) is recommended to avoid amino acid interference when precise NH₄+‐N determinations are needed on dried or fumigated samples.     

A method to assess whether ‘preferential use’ occurs after N ammonium addition; implication for the N isotope dilution technique

The robustness of the assumption of equilibrium between native and added N during ¹⁵N isotope dilution has recently been questioned by Watson et al. (Soil Biol Biochem 32 (2000) 2019-2030). We re-analyzed their raw data using equations that consider the added and native NH₄⁺ and NO₃⁻ pools as separate state variables. Gross mineralization rates and first-order rate constants for NH₄⁺ and NO₃⁻ consumption were obtained by combining analytical integration of the differential equations with a non-linear fitting procedure. The first-order rate constants for NH₄⁺ consumption and NO₃⁻ immobilization for the added NH₄⁺ and NO₃⁻ pool were used to estimate gross mineralization rates and first-order rate constants for nitrification of native NH₄⁺. The latter were 2-4 times lower than the first-order rate constants derived from the added N pool. This discrepancy between first-order rate constants for nitrification implies that one or more process rates estimated for the added N pools cannot be applied to the native N pools. Preferential use of the added N resulted in an overestimation of the gross mineralization by 1.5-2.5-fold, emphasizing the need for critical evaluation of the assumption of equilibrium before gross mineralization rates are calculated.     

Ammonium immobilisation during chloroform fumigation

The assumption in using the chloroform fumigation technique for microbial biomass determination is that microbes are killed or at least inactivated by the treatment. Problems associated with transformations of the N released on or during fumigation have so far only been associated with the fumigation-incubation method. A laboratory and a field study were carried out to investigate the possible N transformations during biomass determination by the fumigation-extraction method. Labelled NH₄NO₃ (either the NO₃⁻, NH₄⁺ or both pools were ¹⁵N enriched) was applied to the soil and biomass determinations made at intervals subsequently. The size and enrichment of the ammonium (NH₄⁺), and nitrate (NO₃⁻) pools were determined before and after chloroform fumigation. The ¹⁵N enrichment of the NH₄⁺ pool after fumigation could only be explained if immobilisation of ammonium occurred at some time during the 24 h fumigation period. The extent of this immobilisation was calculated. In addition, there was evidence that nitrification occurred during the fumigation procedure at the start of the laboratory study and throughout the field study. The laboratory and field study differed mainly in the dynamics related to NO₃⁻ uptake and release. There was evidence for uptake of NO₃⁻ by the microbial biomass with and without utilization. We conclude that the ¹⁵N enrichment in the microbial biomass cannot be accurately determined when N transformations and release of non-utilized N occurs during fumigation. The possible immobilisation of mineral N during fumigation will affect the magnitude of the factor used to convert measured microbial biomass N to actual microbial biomass N in soil.     

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.     

DEVELOPMENT OF A SOIL N TEST FOR FERTILIZER REQUIREMENTS FOR WHEAT

Optimal fertilizer nitrogen (N) rates result in economic yield levels and reduced pollution. A soil test for determining optimal fertilizer N rates for wheat has not been developed for Quebec, Canada, or many other parts of the world. Therefore, the objectives were to determine: 1) the relationship among soil nitrate (NO^? ₃)- N, soil ammonium (NH ⁺ ₄)- N and N fertilizer on wheat yields; and 2) the soil sampling times and depths most highly correlated with yield response to soil NO^? ₃-N and NH ⁺ ₄-N. In a three year research work, wet and dried soil samples of 0- to 30- and 30- to 60-cm depths from 20 wheat fields that received four rates of N fertilizer at seeding and postseeding (plants 15 cm tall) were analyzed for NH ⁺ ₄-N and NO^? ₃ -N using a quick-test (N-Trak) and a standard laboratory method. Wheat yield response to N fertilizer was limited, but strong to soil NO^? ₃-N.     

Arbuscular mycorrhizae affect the N and C economy of nodulated Phaseolus vulgaris (L.) during NH4 nutrition

In the symbiosis between nodulated legume roots and arbuscular mycorrhizal (AM) fungi, the C and N economy can be influenced by the source of N-supply from either AM-derived NH₄⁺ uptake or nodule-derived biological nitrogen fixation (BNF). This relationship was investigated in terms of NH₄⁺ supply and BNF by the two symbionts. Nodulated Phaseolus vulgaris seedlings with and without AM, were hydroponically grown with either 0 N or 1 mM NH₄⁺ supply. Plants were harvested at 30 days after emergence and measurements were taken for biomass, N₂ fixation, photosynthesis, CO₂ and O₂ root respiration, calculated C and N economy. AM roots had higher NH₄⁺ uptake and this was associated with the suppression of BNF and nodule growth. The higher NH₄⁺ uptake in AM roots occurred with lower root maintenance respiration, compared to when N was derived from BNF. There was also an increase in the below-ground sink strength of NH₄⁺ fed AM roots compared to NH₄⁺ fed non-AM roots, as evidenced by the increases in root CO₂ and O₂ respiration and photosynthetic stimulation. These results indicate that although the AM root had higher total below-ground respiratory costs during NH₄⁺ nutrition, there were lower respiratory C costs associated with N derived from AM symbionts in comparison to N from BNF.     

Time-lagged induction of N2O emission and its trade-off with NO emission from a nitrogen fertilized Andisol

Abstract

To understand the influence of basal application of N fertilizer on nitrification potential and N₂O and NO emissions, four soil samples were collected from an upland Andisol field just before (sample 1) and 4 (sample 2), 36 (sample 3) and 72 (sample 4) days after the basal application of N fertilizer during the Chinese cabbage growing season from 12 September to 30 November 2005. The potentials of N₂O production and nitrification of the soils were determined using a ¹⁵N tracer technique and the soils were incubated for 25 days at 25°C and 60% water-filled pore space (WFPS). The results revealed that as much as 84–97% N₂O and almost all NO were produced by nitrification. The ¹⁵N₂O emission peak occurred approximately 350 h after the beginning of incubation for samples 1 and 2, but just 48 h later in samples 3 and 4. Total ¹⁵N₂O emission during the 25-day incubation of samples 3 and 4 ranged from 190 to 198 µg N kg^?1 soil, which was significantly higher than the 99–108 µg N kg^?1 soil recorded in samples 1 and 2. Basal application of N fertilizer did not immediately increase the nitrification potential and the ratio of N₂O to N added, but did dramatically increase the nitrification potential and the ratio of N₂O to N added as (¹⁵NH₄)₂SO₄ 36–72 days after the basal N fertilizer was added. In contrast, NO emission was negatively correlated with nitrification potential and total N₂O emission. As a result, a trade-off relationship between total NO and N₂O emissions was identified. The results indicated that there was a time-lagged induction of the change of N turnover in the soil, which was possibly caused by slow population growth of the nitrifiers and/or a slow shift in the microbial community in the soil.     

Estimates of Gross Transformation Rates of Dairy Manure N Using 15N Pool Dilution

Abstract

Most measurements of dairy manure nitrogen (N) availability depend on net changes in soil inorganic N concentration over time, which overlooks the cycling of manure N in the soil. Gross transformations of manure N, including mineralization (m), immobilization (i), and nitrification (n), can be quantified using ¹⁵N pool dilution methods. This research measures gross m, n, and i resulting from application of four freeze‐dried dairy manures that had distinctly different patterns of N availability. A sandy loam soil (coarse‐loamy, mixed, frigid Typic Haplorthod) was amended with four different freeze‐dried dairy manures and incubated at 25°C with optimal soil water content. The dilution of ¹⁵ammonium (NH4⁺) during a 48‐h interval (7–9 d and 56–58 d after manure application) was used to estimate m, whereas the dilution of ¹⁵nitrate (NO₃ ^?) was used to estimate n. Gross immobilization was calculated as gross minus net mineralization. Gross mineralization in the unamended soil was similar at 7‐ to 9‐d and 56‐ to 58‐d intervals and was significantly increased by the application of manures. For both amended and unamended soil, m was much greater (i.e., three‐ to nine‐fold) than estimated net mineralization, illustrating the degree to which manure N can be cycled in soil. At the early interval, both m and i were directly related to the manure C input, demonstrating the linkage between substrate C availability and N utilization by soil microbes. This research clearly shows that the application of dairy manures stimulates gross N transformation rates in the soil, improving our understanding of the impact of manure application on soil N cycling.