Effect of long-term field exposure of soil to acetylene on nitrification,denitrification, and acetylene consumption

Coated CaC₂ is a newly developed product which can supply nitrification-inhibiting quantities of C₂H₂ (1–10 Pa) to the soil, throughout a cropping season. This method of applying C₂H₂ to the soil maintains C₂H₂ in the soil continuously for several months. It is not know whether these low C₂H₂ concentrations alter soil microbial processes. A field study was initiated to determine the effect of supplying C₂H₂ to a clay soil, using coated CaC₂, on soil respiration, denitrification, nitrification, and C₂H₂ consumption. The C₂H₂ consumption rate increased with length of soil exposure to C₂H₂ (r ²=0.59). The rates of CO₂ production (r ²=0.88) and denitrification (r ²=0.86) were both highly correlated with the C₂H₂ consumption rates. The nitrifier potential decreased to a minimum of 21% of the control after 3 months of C₂H₂ treatment. After this time, nitrifier activity increased to 41% of the control after 11 months of treatment. This increase was due to increased C₂H₂ consumption in the soil. After 3 months of continuous application of C₂H₂ to the soil, the C₂H₂ concentrations were generally below that necessary to inhibit nitrification. No adaptation to the C₂H₂ by nitrifiers was found. Repeating these measurements 1 year later showed that soils previously exposed to C₂H₂ retained their enhanced C₂H₂ oxidation capacity and the capacity to use C₂H₂ to increase denitrification. Nitrification potentials remained about 50% lower in soils exposed to C₂H₂ a year earlier compared to soils not previously exposed to C₂H₂.     

Effect of crop residue C:N ratio on N2O emissions from Gray Lowland soil in Mikasa,Hokkaido, Japan

Abstract

We studied the effect of crop residues with various C:N ratios on N₂O emissions from soil. We set up five experimental plots with four types of crop residues, onion leaf (OL), soybean stem and leaf (SSL), rice straw (RS) and wheat straw (WS), and no residue (NR) on Gray Lowland soil in Mikasa, Hokkaido, Japan. The C:N ratios of these crop residues were 11.6, 14.5, 62.3, and 110, respectively. Based on the results of a questionnaire survey of farmer practices, we determined appropriate application rates: 108, 168, 110, 141 and 0 g C m^?2 and 9.3, 11.6, 1.76, 1.28 and 0 g N m^?2, respectively. We measured N₂O, CO₂ and NO fluxes using a closed chamber method. At the same time, we measured soil temperature at a depth of 5 cm, water-filled pore space (WFPS), and the concentrations of soil NH⁺ ₄-N, NO^? ₃-N and water-soluble organic carbon (WSOC). Significant peaks of N₂O and CO₂ emissions came from OL and SSL just after application, but there were no emissions from RS, WS or NR. There was a significant relationship between N₂O and CO₂ emissions in each treatment except WS, and correlations between CO₂ flux and temperature in RS, soil NH⁺ ₄-N and N₂O flux in SSL and NR, soil NH⁺ ₄-N and CO₂ flux in SSL, and WSOC and CO₂ flux in WS. The ratio of N₂O-N/NO-N increased to approximately 100 in OL and SSL as N₂O emissions increased. Cumulative N₂O and CO₂ emissions increased as the C:N ratio decreased, but not significantly. The ratio of N₂O emission to applied N ranged from ?0.43% to 0.86%, and was significantly correlated with C:N ratio (y = ?0.59 ln [x] + 2.30, r ² = 0.99, P < 0.01). The ratio of CO₂ emissions to applied C ranged from ?5.8% to 45% and was also correlated with C:N ratio, but not significantly (r ² = 0.78, P = 0.11).     

Application of N2/Ar ratio method for N2 fixation studies in submerged soil

Abstract

Recently there has been developments in the measurement of N₂ fixation due mainly to the C₂H₂ reduction method (1). This method, however, has several disadvantages, especially for submerged soil, and the estimated amount of fixed N₂ on the basis of the C₂H₂ reduction activity is not very reliable. The tracer ¹⁵N₂ technique which gives a reliable estimation of the fixed N₂ is too expensive for common use. Development of an alternative method suitable for submerged soil would therefore be desirable. The present authors expected that the measurement of the ratio N₂/Ar in the soil solution might provide advantages for the estimation of the fixed N₂ in submerged soil.     

Saccharidic compounds as soil redox effectors and their influence on potential N2O production

Cellulose, xylan, and glucose were compared in waterlogged soil as modifying factors of the redox potential (Eh), of the quantity of reducing equivalents, and of the soil capacity to produce N₂O and CO₂. During the study period (168 h) soils supplied with glucose and xylan showed a higher Eh decrease than the control soil and the soil treated with cellulose. In samples taken after 0, 24, 48, and 168 h, the soils supplied with C showed a higher number of reducing equivalents than the control soil did. These quantities were not correlated with Eh values, nor with N₂O production. N₂O production was increased compared with the control soil over the entire experimental period in the glucose-amended soils but only after 48 h in the xylan-amended soils and not until 168 h in the cellulose-treated soils. The CO₂:N₂O ratio was consistently higher than the theoretical value of 2, suggesting that denitrification and CO₂ production via fermentation occurred simultaneously. Moreover, this ratio was highly correlated with the Eh values. We conclude that more research is needed to explain the role of soil redox intensity (Eh) and capacity (quantity of redox species undergoing reduction) in the expression of soil denitrification-fermentation pathways.     

The effect of some carbon substrates on denitrification rates and carbon utilization in soil

Summary Soil was amended with a variety of carbon sources, including four soluble compounds (glucose, sucrose, glycerol and mannitol) and two plant residues (straw and alfalfa).. Potential denitrification rates, measured both as N₂O accumulation and NO₃ ^– disappearance, were compared, and the predicted values of available C, measured as CO₂ production and water-extractable C, were assessed.The two measures of denitrification agreed well although N₂O accumulation was, found to be most sensitive. Soil treated with the four soluble C compounds resulted in the same rate of denitrification although glycerol was not as rapidly oxidized. Alfalfa-amended soil produced a significantly higher rate of denitrification than the same amount of added straw. CO₂ evolution was found to be a good predictor of denitrification over the first 2 days of sampling, but neither measure of available substrate C correlated well with denitrification rate beyond 4 days, when NO₃ ^– was depleted in most treatments. The data with alfalfa-amended soil suggested that denitrifiers used water-extractable C. materials produced by other organisms under anaerobic conditions.     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Dynamics of carbon and nitrogen in an extreme alkaline saline soil: A review

The soil of the former lake Texcoco is an ‘extreme’ alkaline saline soil with pH > 10 and electrolytic conductivity (EC) > 150 dS m⁻¹. These conditions have created a unique environment. Application of wastewater sludge to Texcoco soil showed that large amounts of NH₄⁺ were immobilized, NO₃⁻ was reduced aerobically, NO₂⁻ was formed and the mineralization of the organic material in the sludge was inhibited. A series of experiments were initiated to study the processes that inhibited the decomposition of organic material and affected the dynamics of mineral N. The large EC and pH inhibited the decomposition of easily decomposable organic material such as glucose and maize, although cellulolytic activity was observed in soil with pH 9.8 and EC 32.7 dS m⁻¹. The high soil pH favoured NH₃ volatilization of approximately 50 mg N kg⁻¹ soil within a day and a similar amount could be fixed on the soil matrix due to the dispersed minerals and their volcanic origin. Soil microorganisms immobilized large amounts of NH₄⁺ within a day when glucose was added to soil in excess of what was required for metabolic activity. Removal of NO₃⁻ from soil amended with glucose was not inhibited by 100% O₂ and NH₄⁺ indicating that the contribution of denitrification and assimilatory reduction to the reduction of NO₃⁻ was minimal while the formation of NO₂⁻ was not inhibited by 0.1% acetylene, known to inhibit nitrification. Additionally, the reduction of NO₃⁻ in the glucose-amended alkaline saline Texcoco soil was followed by an increase in the amount of NH₄⁺, which could not be due to denitrification. It was concluded that the reduction of NO₃⁻ and the formation of NO₂⁻ and NH₄⁺ in the glucose-amended soil was a result of aerobic NO₃⁻ reduction. A phylogenetic analysis of the archaeal community in the soil of the former lake Texcoco showed that some of the clones identified were capable of reducing NO₃⁻ aerobically to NO₂⁻ when glucose was added. A study of the diversity of the bacterial dissimilatory and respiratory nitrate-reducing communities indicated that bacteria could have contributed to the process.     

Effect of increasing oxygen concentration on total denitrification and nitrous oxide release from soil by different bacteria

Summary The effect of increasing oxygen concentrations (0, 5, 10 and 20 Vol% O₂) on total denitrification and N₂0 release was studied in model experiments using a neutral pH loamy soil relatively rich in easily decomposable organic matter and supplied with nitrate (300 g nitrate N/g dry soil). The sterilized soil was inoculated with three different denitrifying bacteria (Bacillus licheniformis,Aeromonas denitrificans andAzospirillum lipoferum) and incubated (80% WHC, 30°C). The gas volume was analysed for O₂, CO₂, N₂O, NO and N₂ by gas chromatography and the soil investigated for changes in ammonium, nitrite, nitrate, pH, total N and C as well as water-extractable C. WithB. licheniformis andAeromonas denitrificans total denitrification increased remarkably with increasing pO₂ as the result of intensified mineralization.Azospirillum lipoferum, however, showed the highest activity at 5 vol% O₂. WithB. licheniformis N₂O was released only in anaerobic conditions and at 5 Vol% O₂ (maximum) or 10 Vol% 02, but not at 20 Vol%, whereasAeromonas denitrificans produced N₂O only in the presence of He gas (maximum) or at 5 Vol% O₂. In contrast to these bacteria, N₂O production withAzospirillum lipoferum was restricted to 10 Vol% O₂ (maximum) and to 20 Vol% 02, with some traces at 5 vol% O₂. With a certain set of conditions, total denitrification and N₂O formation seem to be governed by the mineralization rate of the organisms in question. The increased demand for electron acceptors by a high turnover rate rather than the presence of anaerobic conditions seems to have determined the rate of denitrification.     

Denitrification,N2-fixation and fermentation during anaerobic incubation of soils amended with glucose and nitrate

Nitrate and glucose additions were investigated for their role in the C and N dynamics during anaerobic incubation of soil. A gas-flow soil core method was used, in which the net production of N₂, N₂O, NO, CO₂, and CH₄ under a He atmosphere could be monitored both accurately and frequently. In all experiments clayey silt loam soil samples were incubated for 9 days at 25 °C. Addition of nitrate (50 mg KNO₃-N kg^-1 soil) had no effect on total denitrification and CO₂ production rates, while the N₂O/N₂ ratio was affected considerably. The cumulative N₂O production exceeded the cumulative N₂ production for 6 days in the treatment with nitrate addition, compared to 1.2 days in the unamended treatment. Glucose addition stimulated the microbial activity considerably. The denitrification rates were limited by the growth rate of the denitrifying population. During denitrification no significant differences were observed between the treatments with 700 mg glucose-C kg^-1 and 4200 mg glucose-C kg^-1, both in combination with 50 mg KNO₃-N kg^-1. The N₂ production rates were remarkably low, until NO _inf3 ^sup- exhaustion caused rapid reduction of N₂O to N₂ at day 2. During the denitrification period 15–18 mg N kg^-1 was immobilised in the growing biomass. After NO _inf3 ^sup- shortage, a second microbial population, capable of N₂-fixation, became increasingly important. This change was clearly reflected in the CO₂ production rates. Net volatile fatty acid (VFA) production was monitored during the net N₂-fixation period with acetate as the dominant product. N₂-fixation faded out, probably due to N₂ shortage, followed by increased VFA production. In the high C treatment butyrate became the most important VFA, while in the low C treatment acetate and butyrate were produced at equal rates. During denitrification no VFA accumulation occurred; this does not prove, however, that denitrification and fermentation appeared sequentially. The experiments illustrate clearly the interactions of C-availability, microbial population and nitrate availability as influencing factors on denitrification and fermentation.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Changes in acetylene reduction activities and effects of inoculated rhizosphere nitrogen-fixing bacteria on rice

Acetylene reduction activities (ARAs) of soils and rice plants during rice-growing season were monitored in temperate region in northeast China. This activity was significantly higher in rhizosphere soil than that in inter-row soil after rice seedlings were transplanted. The ARA was high for most of growing season, suggesting that the native N₂-fixing bacteria responded to rice roots very quickly. Sixteen strains of free-living N₂-fixing bacteria were isolated from three different soils. The ARAs of these strains were correlated with the averaged soil ARAs, suggesting that the isolated strains were likely the active flora responsive to rice roots. The strains were inoculated by soaking seedling roots into the liquid culture for 2 h, and the seedlings were transplanted into pots. Most strains tested did not show any growth-promoting effects except Azotobacter armeniacus and Azotobacter nigricans, which showed growth-promoting effects only at late rice growth stage and only when inoculated in combination but not separately. Present data indicated the promising future applications of these two strains in combination in the region, but further research is needed to understand the underlying mechanisms.     

种植行距和品种对玉米根际反硝化菌群丰度和功能的影响

反硝化是根际氮素损失重要途径,作物品种和行距改变是否会对根际反硝化产生影响尚不清楚。本研究比较了不同玉米品种和种植行距间根际土壤反硝化菌群丰度和功能的差异,为降低根际反硝化损失和提高氮肥利用效率提供科学依据。通过两个独立的田间试验,利用生物化学和分子微生物学方法,分别研究‘浚单20’、‘安农8号’、‘郑单958’、‘品玉18’和‘隆平206’5个玉米品种以及20 cm、30 cm、40 cm、50 cm种植行距对根际土壤反硝化能力、反硝化菌群丰度、N_2O/(N_2O+N_2)产物比和土壤呼吸等指标的影响。‘浚单20’、‘安农8号’、‘郑单958’根际反硝化能力显著低于其他两个品种;随着行距减少,反硝化能力有显著增加趋势。‘隆平206’和‘品玉18’的nir S型反硝化菌群丰度显著高于其他品种,而nir K和nosZ型菌群的丰度以‘浚单20’和‘安农8号’最高;行距20 cm的nir S和nir K型菌丰度显著高于其他行距处理,但nosZ型菌丰度以40 cm行距丰度最大。品种对N_2O/(N_2O+N_2)产物比有一定影响,其中‘安农8号’最低,但行距对产物比没有显著影响。相关分析表明反硝化能力与土壤呼吸和nirS型菌群丰度均极显著正相关,但与nos Z和nir K型菌群未呈现这种关系,由此表明nirS型菌丰度和根际有机碳差异可能是造成反硝化能力不同的主导因子。品种和种植行距会对玉米根际反硝化过程产生一定影响,根际低反硝化损失品种的筛选、选育和根际反硝化过程调控是减少根际反硝化损失,提高氮肥利用效率的有效途径。     

Effect of addition of various carbon substrates on denitrification in a vertic Mollisol

Summary Glucose, acetate, malate, and citrate were added to an agricultural soil. The pe values (-log e^-; calculated from the redox potential) obtained 30 min after the addition of C were not correlated with the theoretical reducing power nor with the theoretical total energy of the C compounds. By contrast the number of electron (e^-) equivalents was correlated with pe⁷, indicating that the proton number affected the redox potential (Eh) measurement. After 24 h of incubation, denitrification rates followed the order citrate>malate>glucose and control. No N₂O production was detected with acetate. Denitrification was not correlated with the theoretical reducing power of the added C compounds but was correlated with pe+pH. Similar numbers of e^- equivalents were measured with all treatments. After 72 h of incubation, the order of the denitrification rates was malate>citrate >acetate>glucose and control. The Eh values (lower than after 24h) did not differ with treatment while the number of e^- equivalents was influenced by the quality of the C source. This also demonstrates that the proton number affected the measured Eh. Our results suggest that the different C substrates did not directly influence the soil physicochemical and biological conditions through their degree of oxidation. Any effects appeared to be indirect, arising from the ability of the substrates to generate new metabolites, and consequently initiate different metabolic pathways that modified the soil physicochemical conditions, reducing power and microbial activity.     

Evaluation of denitrification losses by the acetylene inhibition technique in a permanent ryegrass field (Lolium perenne L.) fertilized with animal slurry or ammonium nitrate

In a field experiment, the effect of animal slurry, (with and without the nitrification inhibitor dicyandiamide on total denitrification losses estimated by the C₂H₂ inhibition technique was measured over 2 years (1989–1990). During this period, four different plots (each with four replicates) were fertilized six times with 150 kg N ha^-1 in the form of cattle-pig slurry or NH₄NO₃. Soil samples (0–20 cm) were analysed at regular intervals for NH _inf4 ^sup+ and NO _inf3 ^sup– concentrations. The soil water content was determined gravimetrically. During the first year (1989) total denitrification losses from unfertilized, mineral-fertilized, and animal slurry-amended plots (with or without dicyandiamide) were estimated as 0.2, 3.1, 0.7, and 0.6 kg N ha^-1, respectively. During the second year (1990) the denitrification losses were 0.4, 1.3, 0.7, and 0.7 kg N ha^-1, respectively. There was a clear relationship between the NO _inf3 ^sup– concentration or soil water content and the denitrification rate. The results are siteund experiment-specific and cannot be generalized so far.     

Use of NaCN to increase the growth of N2-fixing bacteria in a model spermosphere mimicked by sucrose and amino acids leached by seeds

In order to study the establishment of a spermosphere, the C₂H₂ reduction activity of N₂-fixing bacteria isolated from river sand was examined in a simulated spermosphere in the river sand which contained sucrose, an amino acid mixture, and CN^- released from plant seeds. The sand incubated with 10^-10 to 10^-9 mol CN^- 30 g sand^-1 exhibited higher C₂H₂ reduction activity than that without CN^-. The change in the most probable number of N₂ fixers with increasing quantities of CN^- roughly corresponded to that in C₂H₂ reduction activity. However, the most probable number of non-N₂-fixing bacteria decreased except for CN^--tolerant ones. Both C₂H₂ reduction activity and proliferation of the N₂ fixers isolated on a modified Burk's medium were almost similar to those in the bacteria in the sand. In contrast, the proliferation of some nonfixers decreased with an increasing CN^- concentration. C₂H₂ reduction activity of N₂ fixers cultured in combination with non-fixers exhibited a clear peak at 10^-7 M CN^- as for C₂H₂ reduction activity in the sand. We therefore speculate that cyanide evolved from seeds during a pregermination period may suppress the growth of general bacteria, but may promote the proliferation of N₂ fixers, thus contributing to the establishment of a spermosphere.     

Compared effects of long-term pig slurry applications and mineral fertilization on soil denitrification and its end products (N2O, N2)

The long-term (9 years) effect of pig slurry applications vs mineral fertilization on denitrifying activity, N₂O production and soil organic carbon (C) (extractable C, microbial biomass C and total organic C) was compared at three soil depths of adjacent plots. The denitrifying activities were measured on undisturbed soil cores and on sieved soil samples with acetylene method to estimate denitrification rates under field or potential conditions. Pig slurry applications had a moderate impact on the C pools. Total organic C was increased by +6.5% and microbial biomass C by ≥25%. The potential denitrifying activity on soil suspension was stimulated (×1.8, P<0.05) 12 days after the last slurry application. This stimulation was still apparent, but not significant, 10 months later and, according to both methods of denitrifying activity measurement (r ²=0.916, P<0.01 on sieved soil; r ²=0.845, P<0.001 on soil cores), was associated with an increase in microbial biomass C above a threshold of about 105 mg kg⁻¹. The effect of pig slurry on denitrification and N₂O reduction rates was detected on the surface layer (0–20 cm) only. However, no pig slurry effect could be detected on soil cores at field conditions or after NO₃ ⁻ enrichments at 20°C. Although the potential denitrifying activity in sieved soil samples was stimulated, the N₂O production was lower (P<0.03) in the plot fertilized with pig slurry, indicating a lower N₂O/(N₂O + N₂) ratio of the released gases. The pig-slurry-fertilized plot also showed a higher N₂O reduction activity, which is coherent with the lower N₂O production in anaerobiosis.     

Dinitrogen emissions and the N2:N2O emission ratio of a Rendzic Leptosol as influenced by pH and forest thinning

Reduction of nitrous oxide (N₂O) to dinitrogen (N₂) by denitrification in soils is of outstanding ecological significance since it is the prevailing natural process converting reactive nitrogen back into inert molecular dinitrogen. Furthermore, the extent to which N₂O is reduced to N₂ via denitrification is a major regulating factor affecting the magnitude of N₂O emission from soils. However, due to methodological problems in the past, extremely little information is available on N₂ emission and the N₂:N₂O emission ratio for soils of terrestrial ecosystems. In this study, we simultaneously determined N₂ and N₂O emissions from intact soil cores taken from a mountainous beech forest ecosystem. The soil cores were taken from plots with distinct differences in microclimate (warm-dry versus cool-moist) and silvicultural treatment (untreated control versus heavy thinning). Due to different microclimates, the plots showed pronounced differences in pH values (range: 6.3–7.3). N₂O emission from the soil cores was generally very low (2.0 ± 0.5–6.3 ± 3.8 μg N m⁻² h⁻¹ at the warm-dry site and 7.1 ± 3.1–57.4 ± 28.5 μg N m⁻² h⁻¹ at the cool-moist site), thus confirming results from field measurements. However, N₂ emission exceeded N₂O emission by a factor of 21 ± 6–220 ± 122 at the investigated plots. This illustrates that the dominant end product of denitrification at our plots and under the given environmental conditions is N₂ rather than N₂O. N₂ emission showed a huge variability (range: 161 ± 64–1070 ± 499 μg N m⁻² h⁻¹), so that potential effects of microclimate or silvicultural treatment on N₂ emission could not be identified with certainty. However, there was a significant effect of microclimate on the magnitude of N₂O emission as well as on the mean N₂:N₂O emission ratio. N₂:N₂O emission ratios were higher and N₂O emissions were lower for soil cores taken from the plots with warm-dry microclimate as compared to soil cores taken from the cool-moist microclimate plots. We hypothesize that the increase in the N₂:N₂O emission ratio at the warm-dry site was due to higher N₂O reductase activity provoked by the higher soil pH value of this site. Overall, the results of this study show that the N₂:N₂O emission ratio is crucial for understanding the regulation of N₂O fluxes of the investigated soil and that reliable estimates of N₂ emissions are an indispensable prerequisite for accurately calculating total N gas budgets for the investigated ecosystem and very likely for many other terrestrial upland ecosystems as well.     

Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.     

Technique of soil sample pretreatment for analysis of 15N:14N isotope ratio

Abstract

The technique of simultaneous quantitative determination of mineral N soil forms (nitrates, exchangeable and non‐exchangeable ammonium, and total amount of these compounds) and sample pretreatment for the analysis of ¹⁵N:¹⁴N ratio is suggested. The technique is based on the selective association of NH₄ ⁺‐ions into indophenol complex and subsequent ethyl‐acetate extraction of this complex from solution. The mineralization of indophenol is carried out in alkaline medium with simultaneous NH₃ distillation into H₂SO₄ titrant. The application of given technique allows us to shorten significantly the time required for analysis and to increase the accuracy of analytical determination.     

Fate of nitrogen and carbon in the vadose zone: in situ and laboratory measurements of seasonal variations in aerobic respiratory and denitrifying activities

In situ and laboratory measurements of aerobic respiratory and denitrifying activities were studied in the vadose zone (almost 2.5 m thick) of a fluvic hypercalcaric cambisol characterized by transitory anaerobic conditions. A field experiment was conducted in a bare soil, over a 7-month period starting just after maize harvest and incorporation of maize crop residues. Weather variables (air and soil temperature, rainfall), soil water content, soil solutes (NO₃⁻ and dissolved organic carbon) and soil gases (CO₂ and N₂O), were recorded throughout the experiment. Four soil layers were defined. Bacterial counts were performed in each layer using the most probable number (MPN) method. Aerobic respiratory and denitrifying activities were estimated from laboratory measurements. In situ microbial activity, as revealed by CO₂ and N₂O measurements in the soil atmosphere, was strongly influenced by weather. Laboratory measurements showed that potential aerobic respiratory activity (ARA) occurred throughout the soil profile, whereas semi-potential denitrifying activities SPDA (i.e. measured under organic-C limiting condition) occurred mainly in the top 30 cm soil layer. In the soil profile, the CO₂ concentration gradient was stronger than the N₂O concentration gradient. Seasonal variations in microbial activities increased with depth, whereas DOC concentrations, and variations in those concentrations, decreased with depth, suggesting that DOC quality investigations are necessary in the deep vadose zone to understand microbial activities seasonal variations. Laboratory measurements of potential activities agreed well with in situ microbial activity in natural environmental conditions. NO₃⁻ was a stronger limiting factor for SPDA than was denitrifier density in the soil profile.     

The N2O product ratio of nitrification and its dependence on long-term changes in soil pH

The contribution of nitrification to the emission of nitrous oxide (N₂O) from soils may be large, but its regulation is not well understood. The soil pH appears to play a central role for controlling N₂O emissions from soil, partly by affecting the N₂O product ratios of both denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O/(NO₂⁻+NO₃⁻). Mechanisms responsible for apparently high N₂O product ratios of nitrification in acid soils are uncertain. We have investigated the pH regulation of the N₂O product ratio of nitrification in a series of experiments with slurries of soils from long-term liming experiments, spanning a pH range from 4.1 to 7.8. ¹⁵N labelled nitrate (NO₃⁻) was added to assess nitrification rates by pool dilution and to distinguish between N₂O from NO₃⁻ reduction and NH₃ oxidation. Sterilized soil slurries were used to determine the rates of chemodenitrification (i.e. the production of nitric oxide (NO) and N₂O from the chemical decomposition of nitrite (NO₂⁻)) as a function of NO₂⁻ concentrations. Additions of NO₂⁻ to aerobic soil slurries (with ¹⁵N labelled NO₃⁻ added) were used to assess its potential for inducing denitrification at aerobic conditions. For soils with pH?5, we found that the N₂O product ratios for nitrification were low (0.2-0.9‰) and comparable to values found in pure cultures of ammonia-oxidizing bacteria. In mineral soils we found only a minor increase in the N₂O product ratio with increasing soil pH, but the effect was so weak that it justifies a constant N₂O product ratio of nitrification for N₂O emission models. For the soils with pH 4.1 and 4.2, the apparent N₂O product ratio of nitrification was 2 orders of magnitude higher than above pH 5 (76‰ and 14‰). This could partly be accounted for by the rates of chemodenitrification of NO₂⁻. We further found convincing evidence for NO₂⁻-induction of aerobic denitrification in acid soils. The study underlines the role of NO₂⁻, both for regulating denitrification and for the apparent nitrifier-derived N₂O emission.