Klaus Dittert

Contribution of nitrification and denitrification to nitrous oxide emissions from soils after application of biogas waste and other fertilizers

The attribution of nitrous oxide (N(2)O) emission to organic and inorganic N fertilizers requires understanding of how these inputs affect the two biological processes, i.e. denitrification and nitrification. Contradictory findings have been reported when the effects of organic and inorganic fertilizers on nitrous oxide emission were compared. Here we aimed to contribute to the understanding of such variation using (15)N-labelling techniques. We determined the processes producing N(2)O, and tested the effects of soil moisture, N rates, and the availability of organic matter. In a pot experiment, we compared soil treated with biogas waste (BGW) and mineral ammonium sulphate (Min-N) applied at four rates under two soil moisture regimes. We also tested biogas waste, conventional cattle slurry and mineral N fertilizer in a grassland field experiment. During the first 37 days after application we observed N(2)O emissions of 5.6 kg N(2)O-N ha(-1) from soils supplied with biogas waste at a rate of 360 kg N ha(-1). Fluxes were ca. 5-fold higher at 85% than at 65% water holding capacity (WHC). The effects of fertilizer types and N rates on N(2)O emission were significant only when the soil moisture was high. Organic fertilizer treated soils showed much higher N(2)O emissions than those receiving mineral fertilizer in both, pot and field experiment. Over all the treatments the percentage of the applied N emitted as N(2)O was 2.56% in BGW but only 0.68% in Min-N. In the pot experiment isotope labelling indicated that 65-95% of the N(2)O was derived from denitrification for all fertilizer types. However, the ratio of denitrification/nitrification derived N(2)O was lower at 65% than at 85% WHC. We speculate that the application of organic matter in conjunction with ammonium nitrogen first leads to a decrease in denitrification-derived N(2)O emission compared with soil receiving mineral fertilizer. However, at later stages when denitrification becomes C-limited, higher N(2)O emissions are induced when the soil moisture is high.

Novel laser spectroscopic technique for continuous analysis of N2O isotopomers--application and intercomparison with isotope ratio mass spectrometry.

RATIONALE Nitrous oxide (N(2)O), a highly climate-relevant trace gas, is mainly derived from microbial denitrification and nitrification processes in soils. Apportioning N(2)O to these source processes is a challenging task, but better understanding of the processes is required to improve mitigation strategies. The N(2)O site-specific (15)N signatures from denitrification and nitrification have been shown to be clearly different, making this signature a potential tool for N(2)O source identification. We have applied for the first time quantum cascade laser absorption spectroscopy (QCLAS) for the continuous analysis of the intramolecular (15)N distribution of soil-derived N(2)O and compared this with state-of-the-art isotope ratio mass spectrometry (IRMS). METHODS Soil was amended with nitrate and sucrose and incubated in a laboratory setup. The N(2)O release was quantified by FTIR spectroscopy, while the N(2)O intramolecular (15)N distribution was continuously analyzed by online QCLAS at 1 Hz resolution. The QCLAS results on time-integrating flask samples were compared with those from the IRMS analysis. RESULTS The analytical precision (2σ) of QCLAS was around 0.3‰ for the δ(15)N(bulk) and the (15)N site preference (SP) for 1-min average values. Comparing the two techniques on flask samples, excellent agreement (R(2)= 0.99; offset of 1.2‰) was observed for the δ(15)N(bulk) values while for the SP values the correlation was less good (R(2 )= 0.76; offset of 0.9‰), presumably due to the lower precision of the IRMS SP measurements. CONCLUSIONS These findings validate QCLAS as a viable alternative technique with even higher precision than state-of-the-art IRMS. Thus, laser spectroscopy has the potential to contribute significantly to a better understanding of N turnover in soils, which is crucial for advancing strategies to mitigate emissions of this efficient greenhouse gas.

Feedback of grazing on gross rates of N mineralization and inorganic N partitioning in steppe soils of Inner Mongolia

Plant-microbe interactions are crucial regulators of belowground nitrogen cycling in terrestrial ecosystems. However, such interactions have mostly been excluded from experimental setups for the investigation of gross inorganic N fluxes and N partitioning to plants and microorganisms. Ungulate grazing is likely to feed back on soil N fluxes, and hence it is of special importance to simultaneously investigate grazing effects on both plant and microbial N fluxes in intact plant-soil systems, where plant-microbe interactions persist during the experimental incubation. Based on the homogenous 15NH4+ labelling of intact plant-soil monoliths we investigated how various stocking rates (0, 2.35, 4.8 and 7.85 sheep ha−1 grazing season−1) in steppe of Inner Mongolia feedback on gross rates of N mineralization and short-term inorganic N partitioning between plant, microbial and soil N pools. Our results showed that the effect of grazing on gross N mineralization was non-uniform. At low stocking rate gross N mineralization tended to decrease but increased with higher grazing pressure. Hence, there was no significant correlation between stocking rate and gross N mineralization across the investigated grazing intensities. Grazing decreased 15N recovery both in plant and microbial N pools but strongly promoted NO3− accumulation in the soil and thus negatively affected potential ecosystem N retention. This appeared to be closely related to the grazing-induced decline in easily degradable soil C availability at increasing stocking rate.

An RNA Sequencing Transcriptome Analysis Reveals Novel Insights into Molecular Aspects of the Nitrate Impact on the Nodule Activity of Medicago truncatula

A nitrate-induced decline in nitrogenase activity in Medicago truncatula nodules is connected with a strong down-regulation of genes for nodule-specific cysteine-rich peptides and leghemoglobins, changes in inner cell iron allocation, and a decline in nodule respiration efficiency. The mechanism through which nitrate reduces the activity of legume nodules is controversial. The objective of the study was to follow Medicago truncatula nodule activity after nitrate provision continuously and to identify molecular mechanisms, which down-regulate the activity of the nodules. Nodule H2 evolution started to decline after about 4 h of nitrate application. At that point in time, a strong shift in nodule gene expression (RNA sequencing) had occurred (1,120 differentially expressed genes). The most pronounced effect was the down-regulation of 127 genes for nodule-specific cysteine-rich peptides. Various other nodulins were also strongly down-regulated, in particular all the genes for leghemoglobins. In addition, shifts in the expression of genes involved in cellular iron allocation and mitochondrial ATP synthesis were observed. Furthermore, the expression of numerous genes for the formation of proteins and glycoproteins with no obvious function in nodules (e.g. germins, patatin, and thaumatin) was strongly increased. This occurred in conjunction with an up-regulation of genes for proteinase inhibitors, in particular those containing the Kunitz domain. The additionally formed proteins might possibly be involved in reducing nodule oxygen permeability. Between 4 and 28 h of nitrate exposure, a further reduction in nodule activity occurred, and the number of differentially expressed genes almost tripled. In particular, there was a differential expression of genes connected with emerging senescence. It is concluded that nitrate exerts rapid and manifold effects on nitrogenase activity. A certain degree of nitrate tolerance might be achieved when the down-regulatory effect on late nodulins can be alleviated.

Improved diffusion technique for 15N:14N analysis of ammonium and nitrate from aqueous samples by stable isotope spectrometry

Abstract Nitrogen (N) isotope ratio mass spectrometry (IRMS) by Dumas combustion and continuous flow mass spectrometry has become a wide‐spread tool for the studies of N turnover. The speed and labor efficiency of 15N determinations from aqueous solutions such as soil solutions or soil extracts are often limited by sample preparation. Several procedures for the conversion of dissolved ammonium (NH4 +) or nitrate NO3 ‐ to gaseous ammonia and its subsequent trapping in acidified traps have been elaborated in the last decades. They are based on the use of acidified filters kept either above the respective solution or in floating PTFE envelopes. In this paper, we present an improved diffusion method with a fixed PTFE trap. The diffusion containers are continuously kept in a vertical rotary shaker. Quantitative diffusion can thus be achieved in only three days. For solutions with NH4 + levels of only 1 mg N kg‐1 and NO3 ‐ concentrations of 12 mg N kg‐1, recovery rates of 98.8–102% were obtained. By addition of...

RNA-seq transcriptome profiling reveals that Medicago truncatula nodules acclimate N2 fixation before emerging P deficiency reaches the nodules

Summary During a whole-plant P-depletion process in Medicago, formation of new nodules ceases and leaves become P depleted, while existing active nodules maintain high-P levels and display complex molecular acclimation processes.

Soil denitrification potential and its influence on N2O reduction and N2O isotopomer ratios

RATIONALE N2O isotopomer ratios may provide a useful tool for studying N2O source processes in soils and may also help estimating N2O reduction to N2. However, remaining uncertainties about different processes and their characteristic isotope effects still hamper its application. We conducted two laboratory incubation experiments (i) to compare the denitrification potential and N2O/(N2O+N2) product ratio of denitrification of various soil types from Northern Germany, and (ii) to investigate the effect of N2O reduction on the intramolecular (15)N distribution of emitted N2O. METHODS Three contrasting soils (clay, loamy, and sandy soil) were amended with nitrate solution and incubated under N2 -free He atmosphere in a fully automated incubation system over 9 or 28 days in two experiments. N2O, N2, and CO2 release was quantified by online gas chromatography. In addition, the N2O isotopomer ratios were determined by isotope-ratio mass spectrometry (IRMS) and the net enrichment factors of the (15)N site preference (SP) of the N2O-to-N2 reduction step (η(SP)) were estimated using a Rayleigh model. RESULTS The total denitrification rate was highest in clay soil and lowest in sandy soil. Surprisingly, the N2O/(N2O+N2) product ratio in clay and loam soil was identical; however, it was significantly lower in sandy soil. The IRMS measurements revealed highest N2O SP values in clay soil and lowest SP values in sandy soil. The η(SP) values of N2O reduction were between -8.2 and -6.1‰, and a significant relationship between δ(18)O and SP values was found. CONCLUSIONS Both experiments showed that the N2O/(N2O+N2) product ratio of denitrification is not solely controlled by the available carbon content of the soil or by the denitrification rate. Differences in N2O SP values could not be explained by variations in N2O reduction between soils, but rather originate from other processes involved in denitrification. The linear δ(18)O vs SP relationship may be indicative for N2O reduction; however, it deviates significantly from the findings of previous studies.

Influence of Nitrogen Nutrition on Tuber Quality of Potato with Special Reference to the Pathway of Nitrate Transport into Tubers

Abstract The influence of nitrogen (N) nutrition on tuber quality of potato (Solanum tuberosum L.), i.e., specific gravity, starch, nitrate, and protein content as well as ascorbic acid content was investigated in a pot experiment. As expected, specific gravity and starch content decreased with increasing fertilizer rates. Protein and especially nitrate content were positively related to nitrogen fertilization, while the effect on l-ascorbic acid content was relatively small. Only at the highest N level, which led to drastic yield depressions (3.8 g N pot−1), l-ascorbic acid content was significantly reduced. Since is considered phloem-immobile and xylem transport into the tubers is restricted (because of the tubers low transpiration rate), the pathway of nitrate translocation into the tubers is unclear. For Ca2+, a direct uptake from the soil solution by the tuber surface is well documented. In order to check whether this pathway may also be of importance for , potato plants were cultivated in solution culture. The growing tubers were wrapped with paper tissue soaked with Ca(15NO3)2-labelled tuber medium, while contact of the tubers with the nonlabeled liquid growth medium was strictly avoided. 15N-content of tubers was measured on a mass spectrometer. The experiment demonstrated that with high concentration at the tuber surface, up to 40% of total tuber content can be taken up through the tuber skin.

Nitrogen, phosphorus and potassium nutritional status of semiarid steppe grassland in Inner Mongolia

In grazed semiarid steppe ecosystems, much attention has been paid to aspects of growth limitation by water. So far, potential limitation of primary production by plant nutrients was rarely considered. This knowledge is essential for identification of sustainable land-use practices in these large and important ecosystems on the background of over-exploitation and climate change. In the present study plant nutrient concentrations and ratios were investigated with factorial additions of water and N fertilizer at two sites with contrasting soil nutrient availability. Combined analysis of nutrient concentrations, contents, biomass production, and plant N:P ratios consistently confirmed primary growth limitation by water and a strong N limitation when sufficient amounts of water were supplied. P limitation only occurred at the site with low P availability when in addition to the natural supply, water and N fertilizer were given. According to reported thresholds of N:K and K:P ratios, K was not limiting in any plot. The observed nutritional patterns in the plant community were related to the dynamics of species composition and their specific nutrient status. Stipa grandis had the highest N:P ratio whereas Artemisia frigida showed lowest N:P. These nutrient characteristics were related to growth strategies of dominant species. Accordingly, the relative biomass contribution of S. grandis and A. frigida strongly affected the nutrient status of the plant community. Plant N:P ratios indicate the relative limitation by N or P in the semiarid grasslands under sufficient water supply, but other methods of nutritional diagnosis should be used when plant N:P ratios remain below critical values.

Plant responses following grazing removal at different stocking rates in an Inner Mongolia grassland ecosystem

Grazing removal is widely used in grassland management. Plant responses following grazing removal at different organizational levels, however, are not well understood. We examined plant responses at different stocking rates in an Inner Mongolia grassland ecosystem dominated by Leymus chinensis and Stipa grandis. Our results indicated that plant response patterns differed significantly among stocking rates, at different levels of organization, and between wet and dry years. Community aboveground net primary production (ANPP) recovered more quickly at low and moderate stocking rates than those at high stocking rates. Response of aboveground net primary production (RANPP) was significantly positively correlated with both individual biomass and density responses of L. chinensis. Overcompensation of L. chinensis after grazing removal contributed greatly to positive RANPP at the community level. Significant compensatory effects were found between the two dominant species and between dominant species and the remaining non-dominant species. Variation in precipitation significantly affected community ANPP, relationships between community and species responses, and compensatory effects between species. Our study suggests that periodic grazing removal is likely to be a useful method for grassland management and that a combination of species with compensatory effects can be advantageous for reseeding practices in grassland restoration.