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     

Temperature adaptability of soil respiration in short-term incubation experiments

Journal of Soils and Sediments - The soil is the largest carbon pool in the terrestrial ecosystem. Changes of soil respiration with the climate warming are essential for the carbon cycling between...     

Measurement of Hg methylation in sediments using high specific-activity203Hg and ambient incubation

Two methods were developed for estimating the rate ofin situ methylmercury (MeHg) formation in sediments. One method is based on incubation of intact sediment cores without added Hg over a period of days. The second method uses²⁰³HgCl₂ with a specific activity high enough to be used as a tracer (relative to bulk Hg). Use of high-specific activity²⁰³HgCl₂ allowed measurement of methylation rate in hours at ambient total Hg concentrations.²⁰³HgCl₂ was pre-equilibrated with pore water before injection into intact cores, to allow complexation with dissolved ligands. Methylation rates were measured with²⁰³HgCl₂ additions as low as 0.02 μCi and 1.2 ng Hg per g wet weight sediment. These methods were tested in epilimnetic and littoral sediments of two pristine seepage lakes in Northern Wisconsin, and found to compare well.In situ methylation rates in Pallette and Little Rock Lake sediments ranged from 0.1 to 0.4 ug/m² d. Use of²⁰³Hg gave lower errors with shorter incubation times than the ambient incubation method. A method for extraction of Me²⁰³Hg from bulk sediments is given.     

Priming effect of the addition of maize to a Japanese volcanic ash soil and its temperature sensitivity: a short-term incubation study

ABSTRACT

The response of soil organic matter (SOM) to global warming is a crucial subject. However, the temperature sensitivity of SOM turnover remains largely uncertain. Changes in the mineralization of native SOM, i.e., priming effect (PE) may strongly affect the temperature sensitivity of SOM turnover in the presence of global warming. We investigated the direction and magnitude of the PE in a Japanese volcanic ash soil at different temperatures (15°C, 25°C, and 35°C) using a natural ¹³C tracer (C4-plant, maize leaf) in a short-term (25 days) incubation study. In addition, we evaluated the temperature sensitivity expressed as Q₁₀ value with and without the addition of maize to the soil and their relations to PE. We found that positive PE occurred at each temperature condition and tended to increase with decreased temperature, and these PE results were consistent with the microbial biomass at the end of the incubation period. CO₂ emission from control soil (without maize) increased with increasing temperature (Q₁₀ = 2.6), but CO₂ emission from the soil with added maize did not significantly change with increasing temperature (Q₁₀ = 1.0). This was caused by the suppression of CO₂ emission from the soil with increasing temperature (Q₁₀ = 0.9). On the other hand, soil-originated CO₂ emission clearly increased with increasing temperature (Q₁₀ = 3.4) when Q₁₀ values were calculated on the assumption that the temperature and substrate supply increase at the same time (from 25°C). These results suggest that not only the temperature increase but also the labile carbon supply may be important for the temperature sensitivity of Japanese volcanic ash soil.     

厌氧条件下水稻土中铁硫循环与光照的关系

采用恒温厌氧培养试验研究了黑暗、光照、黑暗转光照和光照转黑暗条件下水稻土中硫酸盐还原和铁的氧化还原.结果表明光照是调控土壤铁、硫生物化学转化的一个关键环境因素,光照对铁、硫还原的抑制作用体现在5d后.黑暗培养30 d土壤游离铁的70.07％可被还原;光照培养时35.60 μmol g-1Fe(Ⅲ)先被还原后被氧化,30 d后仍有32.70％的游离铁被还原,转黑暗后被氧化的铁可再次被还原.黑暗时土壤中99.50％的水溶性硫酸盐(WSS)在可5d内被还原,光照培养30 d有42.73％的WSS被还原.水溶性无机碳(WSIC)与体系中Fe(Ⅱ)和WSS之间存在显著相关关系.无光照转换时水溶性有机碳(WSOC)与Fe(Ⅱ)和WSS的转化速率存在显著正相关关系;黑暗条件下WSOC＞ 7.89 μmol g-1时,体系中Fe(Ⅲ)和WSS还原;光照条件下WSOC＞ 8.27 μmol g-1时体系中Fe(Ⅲ)还原,WSOC＞8.40μmol g-1时WSS还原.     

Enhanced anaerobic degradation of hexachlorobenzene in a Hydragric Acrisol using

Humic substances acting as an electron shuttle and nitrogen transformation process influence remarkably the electron transfer in anaerobic reaction systems and thus may affect the reductive dechlorination of hexachlorobenzene(HCB). In order to develop an efficient agricultural strategy for the remediation of organochlorine-contaminated soils, a batch incubation experiment was conducted to study the effects of humic acid, urea, and their interaction on the reductive dechlorination of HCB in a Hydragric Acrisol with high iron oxide content. After 44 d of anaerobic incubation, the five treatments, sterile control,control, humic acid, urea, and humic acid + urea decreased HCB residues by 28.8%, 47.8%, 64.7%, 57.8%, and 71.3%, respectively. The amendment of humic acid or urea significantly decreased soil Eh values and accelerated Fe(Ⅲ) reduction to Fe(Ⅱ), thus promoting markedly reductive dechlorination of HCB. Humic acid had a larger dechlorination effect than urea. Since there was a synergistic interaction between humic acid and urea that accelerated HCB dechlorination, the treatment having both amendments together was the most efficient for HCB dechlorination. The results showed that the combination of NH4⁺_-N supplied by a fertilizer and humic substance is a feasible strategy for the remediation of organochlorine-contaminated soils with abundant iron oxide.     

Genotypic variation for nitrogen uptake and assimilation using hydroponic culture technique in wheat

Hydroponic culture technique is an alternative way of studying nitrogen metabolism. In this study, the response of six wheat genotypes (PBW 621, PBW 636, GLU 1356, BW 8989, GLU 700, and PBW 343) with respect to nitrogen-metabolizing enzymes in relation to accumulation of soluble proteins and amino acids at two concentrations of nitrogen (2 and 6 mM) was studied. Activities of nitrate reductase (NR) and glutamine synthetase as well as soluble proteins, amino acids, and nitrogen content increased in all six genotypes with increasing concentration of nitrogen in roots as well as shoots. Shoots maintained higher activities of NR and glutamine synthetase; apparently contents of soluble protein, amino acid, and nitrogen were also higher. The upregulation of NR and glutamine synthetase activities with increased concentration of nitrogen possibly contributes to higher nitrogen assimilation efficiency of three genotypes (PBW 621, PBW 636, and GLU 1356) compared to other genotypes.     

Contrasting response of nitrification capacity in three agricultural soils to N addition during short-term incubation

Purpose

Human disturbance is a major culprit driving imbalances in the biological transformation of nitrogen from the nonreactive to the reactive pool and is therefore one of the greatest concerns for nitrogen (N) cycling. The objective of this study was to compare potential nitrification rates and the abundance of ammonia oxidizers responsible for nitrification, with the amendment of external N in different agricultural soils.

Materials and methods

Three typical Chinese agricultural soils, QiYang (QY) acid soil, ShenYang (SY) neutral soil, and FengQiu (FQ) alkaline soil, were amended with 0, 20, 150, and 300 μg NH₄ ⁺-N g^?1 soil and incubated for 40 days. The abundance of ammonia oxidizing bacteria (AOB) and archaea (AOA) at the end of incubation in the soil microcosms was determined using the real-time PCR.

Results and discussion

There was a significant decrease in ammonium concentration in the QY soil from the highest to the lowest N-loading treatments, while no significant difference in ammonium concentrations was detected among the different N-loading treatments for the SY and FQ soils. A significantly higher potential nitrification rate (PNR) was observed in the FQ soil while lowest PNR was found in the QY soil. Quantitative PCR analysis of AOB amoA genes demonstrated that AOB abundance was significantly higher in the high N-loading treatments than in the control for the QY soil only, while no significant difference among treatments in the SY and FQ soils. A significant positive correlation between PNR and AOB amoA abundance, however, was found for the SY and FQ soils, but not for the QY soil. Little difference in AOA amoA abundance between different N-loading treatments was observed for all the soils.

Conclusions

This study suggested that ammonia oxidation capacity in the FQ and SY soils was higher than those in the QY soil with the addition of ammonium fertilizer for a short-term. These findings indicated that understanding the differential responses of biological nitrification to varying input levels of ammonium fertilizer is important for maximizing N use efficiency and thereby improving agricultural fertilization management.     

Measurement of penetration force using a Hall-current-sensor

Due to the evident influence of the penetration velocity on the measurement of cone index (CI), diverse motor-operated penetrometers have been designed to keep the velocity with a constant. In this paper, an attractive method to determine the penetration force for a motor-operated penetrometer was presented. Unlike other conventional methods employing various force sensors, this method used a Hall-current-sensor to dynamically measure the operating current of a permanent magnet dc-motor. For the proposed method, both a theoretical analysis related the operating current of the dc-motor to the penetration force and the measurement principle of the Hall-current-sensor were addressed. A soil column with three-layer water contents was used in the laboratory test. The experimental results confirm that the operating current of the dc-motor was a determinate indictor for estimating the penetration force.     

施肥对农田土壤螨季节性变化的影响

对农田土壤螨季节性变化进行试验研究结果表明,农田土壤螨1年有2个发生高峰,即春季5月份高峰和秋季8－9月份高峰。EM堆肥区土壤螨数量年波动平缓,稳定性高;化肥区土壤螨数量年波动性大,中气门目击壤螨7月份后才出现。     

自搅拌厌氧折流板反应器连续处理猪场废水的效果

该研究以猪场废水为处理对象,采用自搅拌厌氧折流板反应器（self-agitation anaerobic baffled reactor,SaABR）开展200 d的连续中温厌氧消化试验,考察在水力停留时间（hydraulic retention time,HRT）3、2、1和0.5 d梯度缩短的过程中,SaABR截留微生物的效果以及反应器的产气性能、稳定性和污泥比产甲烷活性（specific methanogenic activity,SMA）。同时,该研究还开展了全混式反应器（completely stirred tank reactor,CSTR）78 d的连续对比试验。试验发现,SaABR具有良好的截留微生物的作用,在HRT 3 d时SaABR第1至第4取样口污泥挥发性固体（volatile solid,VS）浓度分别为10.2、4.1、44.2和2.5 g/L,而CSTR污泥VS质量浓度仅为2.6 g/L。较高的微生物量显著提高了有机物的降解率并降低了出水的有机酸浓度。随着HRT的缩短,SaABR的降解率也呈现下降。在HRT 1 d时,SaABR的单位VS产甲烷率为0.43 L/g,即使在HRT 缩短到0.5 d时,仍然可实现稳定的发酵产气（单位VS产甲烷率为0.24 L/g）,而CSTR反应器由于微生物洗出不能在HRT 1 d时连续产气。该研究的结果显示,SaABR反应器所具有截留微生物的良好特性,为养殖粪水的处理提供参考。     

Topographic variation in heterotrophic and autotrophic soil respiration in a tropical seasonal forest in Thailand

Soil respiration is a carbon flux that is indispensable for determining carbon balance despite variations over time and space in forest ecosystems. In Kanchanaburi, western Thailand, we measured the soil respiration rates at different slope positions—ridge (plot R), upper slope (plot U), and lower slope (plot L)—on a hill in a seasonal tropical forest [mixed deciduous forest (MDF)] to determine the seasonal and spatial variations in soil respiration on the slope. The heterotrophic (organic layer and soil) and autotrophic (root) respiration was differentiated by trenching. Soil respiration rates showed clear seasonal patterns: high and low rates in rainy and dry seasons respectively, at all plots, and tended to decrease up the slope. Soil respiration rates responded significantly to soil water content in the 0–30?cm layer, but the response patterns differed between the lower slope (plot L) and the upper slope (plots R and U): a linear model could be applied to the lower slope but exponential quadratic models to the upper slope. The annual carbon dioxide (CO₂) efflux from the forest floor was also associated with the slope position and ranged from 1908?gC?m^?2?year^?1 in plot L to 1199?gC?m^?2?year^?1 in plot R. With ascending position from plot L to R, the contribution of autotrophic respiration increased from 19.4 to 36.6% of total soil respiration, while that of the organic layer decreased from 26.2 to 9.4%. Mineral soil contributed to 46.3 to 54.4% of the total soil respiration. Soil water content was the key factor in controlling the soil respiration rate and the contribution of the respiration sources. However, the variable responses of soil respiration to soil water content create a complex distribution of soil respiration at the watershed scale.     

Effects of organic and mineral amendments on available P and phosphatase activities in a degraded Mediterranean soil under short-term incubation experiment

The aim of this study was to determine the effects of mineral and organic-P-fertilizers on soil P availability, bacteria densities and phosphatase activities, in a degraded Mediterranean soil characterized by low level in soil organic matter and nutrients. A typical degraded Mediterranean soil, originating from a siliceous mineral parent material, was amended with different organic or mineral P-sources: aerobically digested sewage sludge (SS), with or without physico-chemical treatment by ferric chloride; sewage sludge compost (SSC); Na or K mineral P-salts (Pi-salts). All the amendments were carried out in order to provide soil with a P total quantity equivalent to 0.5 g P₂O₅/kg of soil. Bacterial density, phosphatase activities (i.e. acid (APH) and alkaline (BPH) phosphomonoesterases and phosphodiesterases), BPH/APH ratio, and available P (P Olsen) were measured after 25 and 87 days of incubation. Results showed that all the P-sources used to fertilize soil during this study resulted in significant increase in P concentration. However, different responses in phosphatase activities and bacterial densities were obtained with regards to the amendment applied to soil. Indeed, it appeared clearly that sewage sludge (SS) considerably stimulated soil biological activity, and more especially the different kinds of phosphatases involved in P mineralization and P turn-over. On the contrary, sewage sludge compost (SSC) as well as P-salts amendments did not affected these parameters in most cases. Results showed also that the incubation time influenced almost all the biological and chemical parameters investigated during this study. As a consequence, P availability was considerably improved in the amended soils between the two sampling dates.     

Factors affecting seasonal variation of methane concentration in water in a freshwater marsh vegetated with Carex lasiocarpa

To understand the underlying factors affecting the seasonal variation of the methane concentration in a cool temperate freshwater marsh vegetated with Carex lasiocarpa in the Sanjiang Plain of northeast China, we measured methane emission from, and the concentrations of methane, dissolved organic carbon (DOC) and acetate in, water samples taken from the standing water surface to the top of the gley soil layer in the C. laisocarpa marsh, before and after plants were covered with a black cloth at the three growing stages of June, July and August 2002. The methane oxidation rate was also measured in situ by applying acetylene, a methane oxidation inhibitor, to whole plants, and the methane production rate in water sampled in June and July was measured via the anaerobic incubation in the laboratory. The methane production rate in water samples was significantly correlated with acetate concentration rather than DOC concentration, whereas the mean acetate concentration in water samples was higher in June than in July and August. Hence, the low methane concentration in June did not result from a lack of acetate for methane production. The mean methane and DOC concentrations in water samples were enhanced by 22.3 and 31.1% in June, 2.1 and 5.0% in July, and 3.4 and 15.2% in August, respectively, after plants were covered with a black cloth. The methane oxidation rate and redox potential in the freshwater marsh decreased from June to July or August. These results suggest that there was more oxygen in the rhizome and rhizosphere in June than in July and August, which not only accentuated methane oxidation but also reduced methane production. Therefore, the high methane concentration in water in July and August could be ascribed to both an increase in temperature and a decrease in redox potential or oxygen concentration in the rhizosphere.     

Denitrification potential in a grassland subsoil: effect of carbon substrates

We examined the potential of a subsoil to denitrify nitrate under optimal anaerobic conditions in a laboratory-based incubation when supplied with a range of C substrates of increasing recalcitrance. Both topsoil and its associated subsoil were supplied with nitrate and either glucose, starch or cellulose. Microbial respiration and the evolution of N₂O and N₂ were measured. The subsoil supported low amounts of microbial activity and responded only to the glucose treatment; with less than one-fifth of the N₂O production measured in the top soil. Overall, our findings demonstrated that the denitrification potential of this particular subsoil is relatively low and that only simple carbohydrates could be utilised readily by the resident microorganisms.     

Modelling in situ activities of enzymes as a tool to explain seasonal variation of soil respiration from agro-ecosystems

Understanding in situ enzyme activities could help clarify the fate of soil organic carbon (SOC), one of the largest uncertainties in predicting future climate. Here, we explored the role of soil temperature and moisture on SOM decomposition by using, for the first time, modelled in situ enzyme activities as a proxy to explain seasonal variation in soil respiration. We measured temperature sensitivities (Q₁₀) of three enzymes (β-glucosidase, xylanase and phenoloxidase) and moisture sensitivity of β-glucosidase from agricultural soils in southwest Germany. Significant seasonal variation was found in potential activities of β-glucosidase, xylanase and phenoloxidase and in Q₁₀ for β-glucosidase and phenoloxidase activities but not for xylanase. We measured moisture sensitivity of β-glucosidase activity at four moisture levels (12%–32%), and fitted a saturation function reflecting increasing substrate limitation due to limited substrate diffusion at low water contents. The moisture response function of β-glucosidase activity remained stable throughout the year. Sensitivity of enzymes to temperature and moisture remains one of the greatest uncertainties in C models. We therefore used the response functions to model temperature-based and temperature and moisture-based in situ enzyme activities to characterize seasonal variation in SOC decomposition. We found temperature to be the main factor controlling in situ enzyme activities. To prove the relevance of our modelling approach, we compared the modelled in situ enzyme activities with soil respiration data measured weekly. Temperature-based in situ enzyme activities explained seasonal variability in soil respiration well, with model efficiencies between 0.35 and 0.78. Fitting an exponential response function to in situ soil temperature explained soil respiration to a lesser extent than our enzyme-based approach. Adding soil moisture as a co-factor improved model efficiencies only partly. Our results demonstrate the potential of this new approach to explain seasonal variation of enzyme related processes.     

利用植物体细胞无性系变异技术选育高粱恢复系

试验研究以高粱恢复系“0-30”幼穗作外植体,用10Gy60Co辐照处理,离体培养得到89株再生苗(R0)。R1代株系27.9%出现7种可见性状的变异,从第2代(R1)中筛选出稳定或分离的优良变异株,经多代选择最终选育出高粱恢复系“011”,用其配制的杂交种“7050A/011”产量高且抗病性强。     

Bacteria sizes in a chernozem as recorded in the course of the microbial succession with the soil incubation under aerobic and anaerobic conditions

The size of the bacteria in different layers and horizons of an arable typical chernozem in the course of the microbial succession under aerobic and anaerobic conditions of incubation was studied. Bacteria with sizes of ??0.23 and ??0.38 ??m predominated under the aerobic conditions; in the anaerobic conditions, their sizes were ??0.17, ??0.23, and ??0.38 ??m. Thus, under anaerobic conditions, the number of bacteria becomes greater at the expense of the increase of their fine forms. The bacterial biomass was the highest (16 ??g/g soil) in the B horizon under the anaerobic conditions, probably, due to the greater number of fine bacterial forms developing in these conditions in nature. In the A horizon under the anaerobic conditions, the bacterial biomass was somewhat lower (12 ??g/g soil) probably because of the high bacterial diversity there. All the soil bacteria were proved to have a size ranging within ??0.43-0.17 ??m.