Transformations of chemically fixed liquid anhydrous ammonia by soil microorganisms

Summary The application of liquid anhydrous NH₃ to soil leads to chemical fixation of NH₃ by organic matter and of NH _inf4 ^sup+ by clay minerals. A laboratory study was conducted to ascertain the biological transformations of newly fixed liquid anhydrous ¹⁵NH₃ in a Drummer silty clay loam by incubation of the ¹⁵N-labelled soil with glucose for 0, 7, 30, and 90 days and by sequential extraction of organic-matter-fixed ¹⁵NH₃ with 0.15 M Na₄P₂O₇, 0.15 M KOH, 0.1 M NaOH, and acidified dimethyl sulfoxide. About 16% of the ¹⁵NH₃ injected was fixed, of which 52% was accounted for by clay fixation. The various humic fractions (fulvic acid, humic acid, and humin) were obtained, and the distribution patterns of the fixed ¹⁵NH₃-N in these fractions were compared. The potential availability of the fixed ¹⁵NH₃-N was also estimated. The percentage of the ¹⁵NH₃ recovered as organic-matter-fixed ¹⁵NH₃ decreased as the length of incubation increased (to 28% after 90 days); the decrease was attributed in part to an increase in the amount recovered as clay-fixed NH _inf4 ^sup+ (from 52 to 64%). Changes in the distribution of the organic-matter-fixed ¹⁵NH₃-N in the humic fractions included: (1) an increase in the relative amount of the fixed ¹⁵NH₃ as humic acid in both the Na₄P₂O₇ and KOH extracts, (2) an increase in the percentage of organic-matter-fixed ¹⁵NH₃-N in the fulvic acid fractions as high-molecular-weight components (determined by dialysis) or as generic fulvic acid (determined by sorption-desorption from XAD-8 resin), and (3) an increase in the percentage of the organic-matter-fixed ¹⁵NH₃ as humin. The potential availability of the organic-matter-fixed ¹⁵NH₃-N decreased as the length of the incubation increased, from 22 to 4% over the 90-day incubation period, and was correlated significantly (0.05 level) with Na₄P₂O₇-extractable N. These results suggest that organic-matter-fixed liquid anhydrous NH₃ is initially more labile than the native soil N but becomes less labile with time.     

黄绵土与淋溶土含水量与施肥方法对铵固定和肥料氮回收率的影响

Ammonium fixation and the effects of soil moisture and application methods on fertilizer N recovery were investigatedin two soils of Shaanxi Province, China, a Luvisol and an Entisol, through two experiments performed in the laboratoryand in a glass shelter, respectively, by using ammonium bicarbonate (NH4HCO3). The laboratory closed incubationbox experiment was conducted using the Luvisol to study NH fixation rate at soil moisture levels of 10.1%, 22.7% and 35.3% water filled pore space (WFPS). The fixed NH -N increased dramatically to 51% and 66%, 67% and 74%,and 82% and 85% 1, 2 and 36 h after fertilizer incorporation at moisture levels of 10.1% and 22.7% WFPS and 35.3% WFPS, respectively. The rapid NH fixation rates at all moisture levels could help prevent NH losses from ammonia volatilization. In the glass shelter pot experiment, N fertilizer was applied by either banding (in a concentrated strip)or incorporating (thoroughly mixing) with the Entisol and the Luvisol. An average of 74.2% of the added N fertilizerwas recovered 26 days after application to the Luvisol, while only 61.4% could be recovered from the Entisol, due tohigher NH fixation capacity of the Luvisol. The amount of fixed NH decreased with increasing WFPS. The amountof fixed NH in the incorporated fertilizer treatment was, oll average, 10% higher than that in the banded treatment.Higher NH fixation rates could prevent N loss and thus increase N recovery. The results from the Luvisol showed lowernitrogen recovery as soil moisture level increased, which could be explained by the fact that most of the fixed NH wasstill not released when the soil moisture level was low. When the fertilizer was incorporated into the soil, the recovery ofN increased, compared with the banded treatment, by an average of 26.2% in the Luvisol and 11.2% in the Entisol, whichimplied that when farmers applied fertilizer, it would be best to mix it well with the soil.     

Incorporation of nitrogen from urea fertilizer into soil organic matter in rice paddy and cassava upland fields in Indonesia

Incorporation of newly-immobilized N into major soil organic matter fractions during a cropping period under paddy and upland cropping systems in the tropics was investigated in Jawa paddy fields with and without fish cultivation and a Sumatra cassava field in Indonesia. ¹⁵N-labelled urea (¹⁵N urea) was applied as basal fertilizer, and the soil samples were collected after harvest. The percentage of distribution of the residual N in soil from ¹⁵N urea into the humic acids, fulvic acid fraction, and humin were 13.1–13.9, 19.0–20.5, and 53.4–54.3%, respectively, for the Jawa paddy soils, and 14.9, 27.4, and 52.4%, respectively, for the Sumatra cassava soil. These values were comparable to the reported ones for other climatic zones. The percentage of distribution of ¹⁵N urea-derived N into humic acids was larger than that of total N into the same fraction in all the soils. The distribution into the fulvic acid fraction was also larger for ¹⁵N urea-derived N than for total N in the Jawa soils. Humic and non-humic substances in the fulvic acid fraction were separated using insoluble polyvinylpyrrolidone (PVP) into the adsorbed and non-adsorbed fractions, respectively. Less than 5% of the ¹⁵N urea-derived N in fulvic acid fraction was detected in the PVP-adsorbed fraction (generic fulvic acids). The proportion of non-hydrolyzable N remained after boiling with 6 M HCl in the ¹⁵N urea-derived N was 9.4–13.5%, 17.3–26.7%, and 8.4–16.6% for the humic acids, generic fulvic acids, and humin, respectively. The significantly low resistance to acid hydrolysis suggested that the ¹⁵N urea-derived N was less stable than the total N in soil regardless of the fractions of humus.     

Temporal changes in distribution and composition of N from labeled fertilizer in soil organic matter fractions

Variations in the amount and composition of immobilized nitrogen (N) in major soil organic matter fractions were investigated in a 730-day soil incubation experiment using ¹⁵N-labeled urea and ¹⁵N nuclear magnetic resonance spectroscopy with the cross polarization/magic angle spinning (¹⁵N CPMAS NMR) method. After 730 days, 24.7% of the applied N was recovered from the soil as organic N. The urea-derived N recovered from humic acids and humin decreased from 11.2 and 33.8% of the applied amount after 14 days to 1.6 and 20.4% after 730 days, respectively. When these values were corrected for the microbial biomass (MB) N, they ranged from 9.0 to 1.2% and 28 to 18%, respectively. The proportion of urea-derived N recovered from fulvic acids was low, ranging between 0.4 and 5.8% (with MB N) or 5.6% (without MB N) of the applied amount, whereas that from water-soluble nonhumic substances (WS-NHS; NHS in the fulvic acid fraction) remained high, 28–33% of the applied amount after correction for the contribution of MB N up to day 365, and decreased to 0.9% thereafter. The ¹⁵N CPMAS NMR spectra of humic acids, fulvic acids, and humin showed the largest signal at −254 to −264 ppm, corresponding to peptide/amide N. The proportions of heterocyclic, peptide/amide, guanidine/aniline, and free amino N in the urea-derived humic acid N were 3–7, 83–90, 5–7, and 2–4%, respectively. More than 80% loss of the urea-derived humic acid N did not markedly alter their composition. No time-dependent variations were also observed for the proportions of respective N functional groups in humin N, which were 3–5, 71–78, 12–17, and 6–10% in the same order as above. These results suggest the greater importance of physical stability than structural variation for the initial accumulation of organic N in soil.     

中国陕西省施有机肥黄土NH4+固定的热力学性质

Some thermodynamic properties of NH₄⁺ fixation by loess soil in plowing and clay layers are discussed. The results indicate that the four ion adsorption equations commonly used can describe the properties of NH₄⁺ fixation in these soils under constant temperature. Among the four adsorption equations, the single-surface Langmuir equation is the best. When the concentration of NH₄Cl solution is 10^-1 mol below, the Freundlich equation can be used. The changes of apparent standard free energy (ΔG°), enthalpy (ΔH°) and entropy (ΔS°) illustrate that NH₄⁺ fixation in soil is an endothermic adsorption and spontaneous reaction, and the process can be enhanced by a higher temperature and clay content in soil. The "proper value of NH₄⁺ fixation by soil (K₁ × q_m) increased with increasing clay content and temperature. The heat of NH₄⁺ fixation in soil (Q_m) confirms the conclusions made in this paper.     

Mineralization of soil organic nitrogen solubilized by aqua ammonia fertilizer

Organic N solubilized by NH_3(aq) was extracted from ¹⁵N-labelled or unlabelled soil, concentrated and added to non-extracted soil, which was incubated under aerobic conditions at 27±1°C. Gross N mineralization, gross N immobilization, and nitrification in soils with or without addition of unlabelled soluble organic N were estimated by models based on the dilution of the NH ₄ ⁺ or NO _inf3 ^sup- pools, which were labelled with ¹⁵N at the beginning of incubation. Mineralization of labelled organic N was measured by the appearance of label in the mineral N pool. Although gross N mineralization and gross N immobilization were increased in two soils between day 0 and day 7 following addition of unlabelled organic N solubilized by NH_3(aq), there was no increase in net N mineralization. Solubilization of ¹⁵N-labelled organic N increased and the ¹⁵N enrichment of the soluble organic N decereased as the concentration of NH_3(aq) added increased. A constant proportion of approximately one-quarter of the labelled organic N added at different rates to non-extracted soil was recovered in the mineral N pool after an incubation period of 14 days, and the availability ratios calculated from net N mineralization data were 1.1:1 and 2.1:1 for 111 and 186 mg added organic-N kg^-1 soil, respectively, indicating that the mineralization of organic N was increased by solubilization.     

Changes in forest soil organic matter pools after a decade of elevated CO2 and O3

The impact of rising atmospheric carbon dioxide (CO₂) may be mitigated, in part, by enhanced rates of net primary production and greater C storage in plant biomass and soil organic matter (SOM). However, C sequestration in forest soils may be offset by other environmental changes such as increasing tropospheric ozone (O₃) or vary based on species-specific growth responses to elevated CO₂. To understand how projected increases in atmospheric CO₂ and O₃ alter SOM formation, we used physical fractionation to characterize soil C and N at the Rhinelander Free Air CO₂-O₃ Enrichment (FACE) experiment. Tracer amounts of ¹⁵NH₄⁺ were applied to the forest floor of Populus tremuloides, P. tremuloides-Betula papyrifera and P. tremuloides-Acer saccharum communities exposed to factorial CO₂ and O₃ treatments. The ¹⁵N tracer and strongly depleted ¹³C-CO₂ were traced into SOM fractions over four years. Over time, C and N increased in coarse particulate organic matter (cPOM) and decreased in mineral-associated organic matter (MAOM) under elevated CO₂ relative to ambient CO₂. As main effects, neither CO₂ nor O₃ significantly altered ¹⁵N recovery in SOM. Elevated CO₂ significantly increased new C in all SOM fractions, and significantly decreased old C in fine POM (fPOM) and MAOM over the duration of our study. Overall, our observations indicate that elevated CO₂ has altered SOM cycling at this site to favor C and N accumulation in less stable pools, with more rapid turnover. Elevated O₃ had the opposite effect, significantly reducing cPOM N by 15% and significantly increasing the C:N ratio by 7%. Our results demonstrate that CO₂ can enhance SOM turnover, potentially limiting long-term C sequestration in terrestrial ecosystems; plant community composition is an important determinant of the magnitude of this response.     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Comparison of 15N natural abundance and difference methods for estimating N2 fixation of soybeans grown in a tropical soil

Abstract

A study was carried out to compare the difference or N-yield method with the ¹⁵N natural abundance method for the estimation of the fractional contribution of biological N₂ fixation in the different plant parts of nodulating and non-nodulating isolines of soybeans. The results indicated that the δ¹⁵N values of most plant parts of soybeans were significantly lower (p<0.05) in the nodulating than in the non-nodulating isoline. However, in the case of the root+nodule component, the δ¹⁵N value was higher in the nodulating than in the non-nodulating isoline possibly due to isotopic discrimination of ¹⁵N over ¹⁴N which may have occurred in the nodules. Inoculation of soybeans with the Bradyrhizobium japonicum strain CB 1809 increased significantly (p<0.05) the δ¹⁵N value of the root+nodule component implying that the effectiveness of the soybean-rhizobium symbiosis had increased by inoculation.

Percentage of plant N derived from atmospheric N₂ fixation (%Ndfa) estimated by the ¹⁵N natural abundance method was highly correlated (r=0.762, p<0.01) with that by the difference or N-yield method and the differences between the two methods were not statistically significant. The agreement between the two methods was closer at maturity than at the early reproductive stage.

The %Ndfa obtained by the difference method ranged from 48.4 to 92.6% whereas the %Ndfa obtained by the ¹⁵N natural abundance method ranged from 43.2 to 92.4% in the different plant parts. Based on the ¹⁵N natural abundance method, approximately 15% of the N in pod, shoot, grain, and shell was derived from the soil but in the case of stover, this fraction was about 55%.     

C and N dynamics of a range of biogas slurries as a function of application rate and soil texture: a laboratory experiment

Digestates vary in composition and studies regarding their impact on C and N dynamics in soils are scarce. The objective was to analyse the C and N dynamics of digestates originating from various substrates applied to a sandy Cambisol and a silty Anthrosol. In three laboratory experiments (4–6 weeks), the effects of digestate properties, N rate and water content were tested. Averaged over both soils, 21% of the C supplied was emitted as CO₂. Potential NH₃ emissions during the first week ranged between 6% and 12% of NH₄⁺ present in the digestates. The emission factors in the sandy Cambisol were on average 1.2 and 2 times higher for CO₂ and potential NH₃, respectively, compared to the silty Anthrosol. Similarly, net nitrogen mineralization in the sandy Cambisol was approximately twice the N mineralized in the silty Anthrosol. Net nitrification was not influenced by soil texture or different digestates, but increased with increasing application rates and had highest values at 75% of water holding capacity. Our results indicate that the type of substrate input for anaerobic digestion influences the properties of the digestate and therefore the dynamics of C and N. However, soil texture can affect these dynamics markedly.     

Nitrogen-15 balance in transplanted and direct-seeded flooded rice as affected by different methods of urea application

Summary Alternative N-fertilizer management practices are needed to increase productivity and the N-use efficiency of flooded rice (Oryza sativa L.). In the 1987 dry season, a field study using ¹⁵N-labeled urea evaluated the effect of the time and method of fertilizer-N application on grain yield and N-use efficiency in transplanted and direct-seeded flooded rice. Conventional fertilizer application (broadcasting and incorporation) was compared with band placement of liquid urea and point placement of urea supergranules. With band or point placement, the grain yields were significantly greater, and the partial pressure of NH₃ (pNH₃) in the floodwater was significantly reduced. In the transplanted rice, conventional fertilizer-N application gave a 64% total ¹⁵N recovery and 38% crop (grain and straw) recovery. Band placement of liquid urea N resulted in 92% total and 73% crop recovery. In the direct-seeded flooded rice, a conventional N application gave 72% total and 42% crop recovery; band placement, 98% total and 73% crop recovery; and urea supergranule point placement, 97% total and 75% crop recovery.Dedicated to Prof. Dr. K. Mengel, Giessen, FRG, in honor of his 60th birthday.     

Nitrapyrin effectiveness in reducing nitrous oxide emissions decreases at low doses of urea in an Andosol

In the tropics,frequent nitrogen(N)fertilization of grazing areas can potentially increase nitrous oxide(N₂O)emissions.The application of nitrification inhibitors has been reported as an effective management practice for potentially reducing N loss from the soil-plant system and improving N use efficiency(NUE).The aim of this study was to determine the effect of the co-application of nitrapyrin(a nitrification inhibitor,NI)and urea in a tropical Andosol on the behavior of N and the emissions of N₂O from autotrophic and heterotrophic nitrification.A greenhouse experiment was performed using a soil(pH 5.9,organic matter content 78 g kg^-1,and N 5.6 g kg^-1)sown with Cynodon nlemfuensis at 60%water-filled pore space to quantify total N₂O emissions,N₂O derived from fertilizer,soil ammonium(NH₄⁺)and nitrate(NO₃^-),and NUE.The study included treatments that received deionized water only(control,NI).No significant differences were observed in soil NH₄⁺content between the UR and UR+NI treatments,probably because of soil mineralization and NO₃^-produced by heterotrophic nitrification,which is not effectively inhibited by nitrapyrin.After 56 d,N₂O emissions in UR(0.51±0.12 mg N₂O-N concluded that the soil organic N mineralization and heterotrophic nitrification are the main processes of NH₄⁺and NO₃^-production.Additionally,it was found that N₂O emissions were partially a consequence of the direct oxidation of the soil's organic N via heterotrophic nitrification coupled to denitrification.Finally,the results suggest that nitrapyrin would likely exert significant mitigation on N₂O emissions only if a substantial N surplus exists in soils with high organic matter content.     

Dynamics of microbial biomass nitrogen as influenced by organic matter application in paddy fields

The effects of annual application of rice straw or cow manure compost for 17–20 y on the dynamics of fertilizer N and soil organic N in Gley paddy fields were investigated by using the ¹⁵N tracer technique during the rice cropping season. The chloroform fumigation-extraction method was evaluated to determine the properties of soil microbial biomass under submerged field conditions at the tillering stage before mid-summer drainage, with special reference to the fate of applied NH₄ ⁺-¹⁵N.

The transfer ratios from applied NH₄ ⁺-¹⁵N to immobilized N in soil and to uptake N by rice during given periods varied with the rice growth stages and were affected by organic matter application. The accumulated amounts of netmineralized soil organic N (net-M_j ), immobilized N (I_j ), and denitrified N (D_j ) during the cropping season were estimated to be 14.0–22.5, 6.3–11.2, and 3.4–5.3 g N m^-2, respectively. Values of net-M_j and I_j were larger in the following order: cow manure compost plot > rice straw plot > plot without organic matter application, and their larger increase by the application of cow manure compost contributed to a decrease of the D_j values, as compared with rice straw application.

Values of E _N extra extractable soil total N after fumigation, increased following organic matter application, ranging from 2.1 to 5.4 g N m^-2. Small residual ratios of applied ¹⁵N in the fraction E _N at the end of the given period indicated that re-mineralization of newly-assimilated ¹⁵N through the easily decomposable fraction of microbial biomass had almost ended. Thus, the applicability to paddy field soils of the chloroform fumigation-extraction method was confirmed.     

Short and long-term effects of elevated CO2 on Lolium perenne rhizodeposition and its consequences on soil organic matter turnover and plant N yield

It is still unclear whether elevated CO₂ increases plant root exudation and consequently affects the soil microbial biomass. The effects of elevated CO₂ on the fate of the C and nitrogen (N) contained in old soil organic matter pools is also unclear. In this study the short and long-term effects of elevated CO₂ on C and N pools and fluxes were assessed by growing isolated plants of ryegrass (Lolium perenne) in glasshouses at elevated and ambient atmospheric CO₂ and using soil from the New Zealand FACE site that had >4 years exposure to CO₂ enrichment. Using ¹⁴CO₂ pulse labelling, the effects of elevated CO₂ on C allocation within the plant-soil system were studied. Under elevated CO₂ more root derived C was found in the soil and in the microbial biomass 48 h after labelling. The increased availability of substrate significantly stimulated soil microbial growth and acted as priming effect, enhancing native soil organic matter decomposition regardless of the mineral N supply. Despite indications of faster N cycling in soil under elevated CO₂, N availability to plants stayed unchanged. Soil previously exposed to elevated CO₂ exhibited a higher N cycling rate but again there was no effect on plant N uptake. With respect to the difficulties of extrapolating glasshouse experiment results to the field, we concluded that the accumulation of coarse organic matter observed in the field under elevated CO₂ was probably not created by an imbalance between C and N but was likely to be due to more complex phenomena involving soil mesofauna and/or other nutrients limitations.     

Arbuscular mycorrhizal fungi contribute to C and N enrichment of soil organic matter in forest soils

Increasing evidence suggests that accretion of microbial turnover products is an important driver for isotopic carbon (C) and nitrogen (N) enrichment of soil organic matter (SOM). However, the exact contribution of arbuscular mycorrhizal fungi (AMF) to soil isotopic patterns remains unknown. In this study, we compared ¹³C and ¹⁵N patterns of glomalin-related soil protein (GRSP), which includes a main fraction derived from AMF, litter, and bulk soil in four temperate rainforests. GRSP was an abundant C and N pool in these forest soils, showing significant ¹³C and ¹⁵N enrichment relative to litter and bulk soil. Hence, cumulative accumulation of recalcitrant AMF turnover products in the soil profile likely contributes to ¹³C and ¹⁵N enrichment in forest soils. Further research on the relationship between GRSP and AMF should clarify the exact extent of this process.     

Contribution of red alder to soil nitrogen input in a silvopastoral system

Studies on Alnus rubra Bong. (red alder) were carried out to assess its potential for use as a component of a silvopastoral system. Comparison was between two treatments: red alder at 400 stems ha⁻¹ (silvopasture) and red alder at 2,500 stems ha⁻¹ (forestry control). δ¹⁵N values close to zero were recorded in all red alder plant parts except for root nodules, indicating that a large proportion of N in red alder was fixed from the atmosphere. Overall, it was estimated that there was 63.45 kg N ha⁻¹ fixed N accumulated in red alder trees, and the rate of N fixation was estimated at 30.95 kg ha⁻¹ year⁻¹ in the silvopasture treatment. The total amount of N that could potentially be added to the soil in the silvopasture treatment as a result of decomposition of senescent leaves, roots, and dead nodules was estimated at 40.56 kg ha⁻¹ year⁻¹. Of the total N added to the soil, 27.1 kg ha⁻¹ year⁻¹ was due to N fixation from the atmosphere. These results show that red alder has a potential to improve and maintain soil fertility in a silvopastoral system.     

Gross N mineralization rates after application of composted grape marc to soil

Grape marc is a common waste product of the wine production industry. When partially composted and applied to soil it may contain enough N to affect vine growth and hence wine quality. Yet little is known about the quantity and timing of N release from composted grape marc. A laboratory incubation was conducted where composted grape marc amended and non-amended soils were periodically sampled over 148 days at 15 °C for gross N mineralization rates, C mineralization and microbial biomass-C. Gross N mineralization rates were determined by ¹⁵N pool dilution using both analytical equations and the numerical model FLUAZ (Mary, B., Recous, S., Robin, D., 1998. A model for calculating nitrogen fluxes in soil using 15N tracing. Soil Biology & Biochemistry 30, 1963-1979.). Both analytical and FLUAZ determined gross N mineralization rates were in close agreement in the control soil. However, in composted grape marc amended soils there was a discrepancy between the two solutions. Findings indicate that composted grape marc caused a net immobilization of N for the first 50-days of incubation, after which enough N was released to require consideration in fertilizer-N strategies.     

Dynamics of 15N-labeled ammonium sulfate in various inorganic and organic soil fractions of wetland rice soils

Summary The dynamics of basally applied ¹⁵N-labeled ammonium sulfate in inorganic and organic soil fractions of five wetland rice soils of the Philippines was studied in a greenhouse experiment. Soil and plant samples were collected and analyzed for ¹⁵N at various growth stages. Exchangeable NH₄ ⁺ depletion continued after 40 days after transplanting (DAT) and corresponded with increased nitrogen uptake by rice plants. Part of the applied fertilizer was fixed by 2:1 clay minerals, especially in Maligaya silty clay loam, which contained beidellite as the dominant clay mineral. After the initial fixation, nonexchangeable ¹⁵N was released from 20 DAT in Maligaya silty clay loam, but fixation delayed fertilizer N uptake from the soil. Part of the applied N was immobilized into the organic fraction. In Guadalupe clay and Maligaya silty clay loam, immobilization increased with time while the three other soils showed significant release of fertilizer N from the organic fraction during crop growth. Most of the immobilized fertilizer N was recovered in the nondistillable acid soluble (alpha-amino acid + hydrolyzable unknown-N) fraction at crop maturity. Between 61% and 66% of applied N was recovered from the plant in four soils while 52% of fertilizer N was recovered from the plant in Maligaya silty loam. Only 20% – 30% of the total N uptake at maturity was derived from fertilizer N. Nmin (mineral N) content of the soil before transplanting significantly correlated with N uptake. Twenty-two to 34% of applied N was unaccounted for possibly due to denitrification and ammonia volatilization.     

Evaluation of N2 fixation by applying 15N labeled plant material and ammonium sulfate

Effect of different ¹⁵N labeled sources on the estimation of N₂ fixation was investigated. The combination of ¹⁵N labeled ammonium sulfate, ¹⁵N labeled plant material, and ¹⁵N labeled ammonium sulfate with unlabeled plant material, was examined in pot experiments. Two cultivars of soybean (Glycine max) and one of mungbean (Vigna radiata) were used. No significant difference was observed among the treatments for the estimation of N₂ fixation. This was due to the homogeneity and stability of the ¹⁵N abundance in soil which resulted in a similar N uptake from the soil by the N² fixing and reference crops. The plant yield, total N uptake and amount of N₂ fixed were higher in the Yellow Soil than in the Andosol. The amount of N₂ fixed was strongly influenced by the plant growth and consequently it affected the plant yield. The slow decomposition of plant material in the Andosol resulted in a low yield in both the N₂ fixing and reference crops. Thus, the artificial decrease of the available N content in soil, by application of plant material, did not stimulate N, fixation but suppressed plant growth and N₂ fixation.     

The contribution of soil organic matter fractions to carbon and nitrogen mineralization and microbial community size and structure

The aims of this study were to: (i) assess the impact of hay and fertilizer application on organic matter (OM) fractions (dissolved organic matter (DOM), light fraction organic matter (LFOM, <1.0 g cm⁻³), heavy fraction OM (HFOM, <1.7 g cm⁻³)), carbon (C) and nitrogen (N) cycling processes and microbial community size and structure, and (ii) quantify the role of OM fractions to C and N cycling. Soil was collected in 2001 from a field experiment to which grass hay (1996) and/or fertilizer (1995 and 1999) had previously been applied. DOM-C (P<0.05) and DOM-N (P=0.07) were significantly higher in control and fertilized soil than hay and hay+fertilized soil. LFOM and HFOM C and N contents and C/N ratios were significantly (P<0.05) higher in hay+fertilized and hay amended soil than in control and fertilized soil. Potentially mineralizable-N (PMN), microbial biomass-C (MB-C), microbial biomass-N (MB-N) and microbial respiration (CO₂) were not affected by fertilizer and/or hay application. Gross N mineralization (Gross Min) and gross nitrification (Gross Nit) rates were significantly (P<0.05) higher in fertilized, hay, hay+fertilized soil than control soil. However, there was no significant difference between treatments in gross N immobilization rates. Results reported here highlight the importance of a labile fraction of the DOM pool to N and C cycling as its removal significantly (P<0.05) reduced PMN, MB-N, Gross Min and Gross Nit compared with whole soil in most or all treatments. In soil where DOM+LFOM were removed PMN was significantly (P<0.05) lower, but MB-C, Gross Min and Gross Nit was significantly (P<0.05) higher than in DOM removed soil. This suggests that LFOM plays an important role as a sink for mineral-N. Total soil phospholipid fatty acid (PLFA) concentration was significantly (P<0.05) higher in hay amended than control, fertilized and hay+fertilized soil. Principal components analysis was able to clearly discriminate between control, fertilized, hay+fertilized and hay amended soil. Soil amended with hay or fertilizer had a microbial community structure which differed from that of the control or hay+fertilized soils. Redundancy analysis with Monte Carlo permutation tests revealed that PLFA profiles were strongly correlated to differences in Gross Min, Gross Nit, MB-N, MB-C, MB-C/N ratio, total soil C and total soil C/N ratio. The results of this research suggest that changes in microbial structure are related to aspects of soil C and N pools and cycling.