Browsing by Subject "Nitrifikation"

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Abundance and diversity of total and nitrifying prokaryotes as influenced by biochemical quality of organic inputs, mineral nitrogen fertilizer and soil texture in tropical agro-ecosystems
(2016) Muema, Esther Kathini; Cadisch, Georg
Tropical agro-ecosystems are limited in nutrient resources as a consequence of i) being composed of highly weathered soils, ii) low native soil organic matter (SOM) content due to conversion of natural forests to arable lands and iii) continuous cropping without replenishing soil nutrients. Recovery of SOM by use of organic residues is faced with other competing uses like animal fodder. Moreover, existing SOM is further reduced by increased turnover rates due to favorable climatic conditions in the tropics. Incorporation of residues is therefore a justified means to restore SOM and to provide crop nutrients through microbial mediated activities like nitrification. Nitrification is a central step of the nitrogen (N) cycle, whereby ammonia is converted into nitrite and then to nitrate by bacteria and archaea through production of the amoA gene encoding the alpha-subunit of the enzyme ammonia monooxygenase. In order to better understand the impact of organic residues of contrasting biochemical quality (i.e., high quality Tithonia diversifolia (TD; C/N ratio: 13, lignin: 8.9 %, polyphenols: 1.7 %), intermediate quality Calliandra calothyrsus (CC; 13, 13, 9.4) and low quality Zea mays (ZM; 59, 5.4, 1.2)) on nutrient provision, effects of residue quality on dynamics of relevant decomposer microbial communities were studied. In addition, mineral N fertilizer was used to compensate for mineral N limitations especially in case of low and intermediate quality residues. Since N is one of the most limiting crop nutrients in the tropics, this study therefore focused on ammonia-oxidizing prokaryotes, using DNA-based quantitative PCR (qPCR) and terminal restriction fragment length polymorphism (TRFLP) techniques. In addition, soil physicochemical properties were measured and linked to the dynamics of microbial communities. The study hypothesized that soil type due to differences in structure and nutrient background, as well as seasonality, which influences soil moisture, would shape the response of the studied communities to biochemical quality of residues. Overall, the results of this PhD research revealed specific responses of dynamics of AOB and AOA to quality of organic residues and their combinations with mineral N fertilizer. They also revealed effects of interrelations between quality of residues and soil texture as well as seasonality particularly precipitation on dynamics of microbial communities. Future investigation of active microbial communities with the use of RNA-based approaches need to be considered to further improve our understanding of quality of SOM on soil nutrient dynamics.
Developing indicators and characterizing direct and residual effects of biological nitrification inhibition (BNI) by the tropical forage grass Brachiaria humidicola
(2018) Karwat, Hannes; Cadisch, Georg
Nitrogen (N) losses from agroecosystems harm the environment via increased nitrate (NO3-) amounts in water-bodies and nitrous oxide (N2O) emissions to the atmosphere. Bacteria and archaea oxidize ammonium (NH4+) to NO3- under aerobic conditions. Furthermore, under mainly anaerobic conditions, microbial denitrification reduces NO3- to gaseous N forms. The tropical forage grass Brachiaria humidicola (Rendle) Schweick (Bh) has been shown to reduce soil microbial nitrification via root derived substances. Therefore, biological nitrification inhibition (BNI) by Bh might contribute to reduction of N losses from agroecosystems. The present doctoral thesis aimed at assessing the potential of the actual BNI by Bh, as well as the residual BNI effect with new developed methodologies. The overall research was based on the following major objectives: (1) characterization of the residual BNI effect by Bh on recovery of N by subsequent cropped maize (Zea mays L.) under different N fertilization rates; (2) investigate if low enzymatic nitrate reductase activity (NRA) in leaves of Bh is linked to reduced NO3- nutrition by effective BNI; (3) identify a possible link between plant delta 15N of Bh and the BNI effect of different Bh genotypes on nitrification, plant N uptake and NO3- leaching losses. The overall objective was to use and test new methodologies with a minimum of disturbance of the plant-soil system, to characterize BNI of different Bh genotypes in greenhouse and field studies. The first research study focused on the investigation of a potential residual BNI effect of a converted long-term Bh pasture on subsequent maize cropping, where a long-term maize monocrop field served as control. The residual BNI effect was characterized in terms of enhanced maize grain yield, total N uptake and 15N (labeled) fertilizer recovery. Furthermore, the impact of residual BNI effect on soil N dynamics was investigated. The residual BNI effect was confirmed for the first maize crop season after pasture conversion on the basis of lower nitrification in incubation soil, higher total N uptake and higher maize grain yields. However, the residual BNI effect did not result in higher 15N fertilizer uptake or reduced 15N fertilizer losses, nor in reduced N20 emissions. Applied N was strongly immobilized due to long-term root turnover effects, while a significant residual BNI effect from Bh prevented re-mineralized N from rapid nitrification resulting in improved maize performance. A significant residual Bh BNI effect was evident for less than one year only. In the second research study it was the aim to verify the potential of nitrate reductase activity (NRA) as a proxy for the detection of in vivo performance of BNI by selected Bh accessions and genotypes grown under contrasting fertilization regimes. NRA was detected in Bh leaves rather than in roots, regardless of NO3- availability. Leaf NRA correlated with NO3- contents in soils and stem sap of contrasting Bh genotypes substantiating its use as a proxy of in vivo performance of BNI. The leaf NRA assay facilitated a rapid screening of contrasting Bh genotypes for their differences in in vivo performance of BNI under field and greenhouse conditions; but inconsistency of the BNI potential by selected Bh genotypes was observed. The third research study emphasized to link the natural abundance of delta 15N in Bh plants with reduced NO3- losses and enhanced N uptake due to BNI. Increased leached NO3- was positively correlated to rising delta 15N in Bh grass, whereas the correlation between plant N uptake and plant delta 15N was inverse. Long-term field cultivation of Bh decreased nitrification in incubated soil, whereas delta 15N of Bh declined and plant N% rose over time. Delta 15N of Bh correlated positively with assessed nitrification rates in incubated soil. It was concluded that decreasing delta 15N of Bh over time reflects the long-term effect of BNI linked to lower NO3- formation and reduced NO3- leaching, and that generally higher BNI activity of Bh is indicated by lower delta 15N plant values. Within the framework of this thesis, a residual BNI effect by Bh on maize cropping could be confirmed for one season due to the combined methodological approaches of soil incubation and 15N recovery. The development of the NRA assay for sampled Bh leaves was validated as a rapid and reliable method linked to the actual soil nitrification after NH4+ fertilizer supply. Consequently, the assay could be used for both greenhouse and field studies as BNI proxy. The gathered data from the third study indicated that decreasing delta 15N of Bh over time reflects the long-term effect of BNI linked to lower NO3- formation and reduced NO3- leaching, and that generally higher BNI activity of Bh is indicated by lower delta 15N plant values. Consequently, it was suggested that delta 15N of Bh could serve as an indicator of cumulative NO3- losses. Overall, this doctoral thesis suggests the depressing effect on nitrification by Bh might be a combined effect by BNI and fostered N immobilization. Furthermore, BNI by Bh might be altered by different factors such as soil type, plant age and root morphology of the genotypes. Finally, future studies should consider that Bh genotypes express their respective BNI potential differently under contrasting conditions.
Linking microbial abundance and function to understand nitrogen cycling in grassland soils
(2017) Regan, Kathleen Marie; Kandeler, Ellen
This thesis characterized spatial and temporal relationships of the soil microbial community, the nitrogen cycling microbial community, and a subset of the nitrogen cycling community with soil abiotic properties and plant growth stages in an unfertilized temperate grassland. Unfertilized perennial grasslands depend solely on soil-available nitrogen and in these environments nitrogen cycling is considered to be both highly efficient and tightly coupled to plant growth. Unfertilized perennial grasslands with high plant diversity, such as ours, have also been shown to have higher soil organic carbon, total nitrogen, and microbial carbon; greater food web complexity; and more complex biological communities than more intensively managed grasslands or croplands. This made the choice of study plot especially well-suited for characterizing the relationships we sought to identify, and made it possible to detect spatial and temporal patterns at a scale that has heretofore been under-examined. The first study used a combination of abiotic, plant functional group, and PLFA measurements together with spatial statistics to interpret spatial and temporal changes in the microbial community over a season. We found that its overall structure was strongly related to the abiotic environment throughout the sampling period. The strength of that relationship varied, however, indicating that it was not constant over time and that other factors also influenced microbial community composition. PLFA analysis combined with principal components analysis made it possible to discern changes in abundances and spatial distributions among Gram-positive and Gram-negative bacteria as well as saprotrophic fungi. Modeled variograms and kriged maps of the changes in distributions of exemplary lipids of both bacterial groups also showed distinct differences in their distributions on the plot, especially at stages of most rapid plant growth. Although environmental properties were identified as the main structuring agents of the microbial community, components of those environmental properties varied over the season, suggesting that plant growth stage had an indirect influence, providing evidence of the complexity and dynamic nature of the microbial community in a grassland soil. The second study took the same analytical approach, this time applying it to abundances of key members of the soil nitrogen cycling community. Marker genes for total archaea and bacteria, nitrogen fixing bacteria, ammonia oxidizing archaea and bacteria, and denitrifying bacteria were quantified by qPCR. Potential nitrification activity and denitrifying enzyme activity were also determined. We found clear seasonal changes in the patterns of abundance of the measured genes and could associate these with changes in substrate availability related to plant growth stages. Most strikingly, we saw that small and ephemeral changes in soil environmental conditions resulted in changes in these microbial communities, while at the same time, process rates of their respective potential enzyme activities remained relatively stable. This suggests both short term niche-partitioning and functional redundancy within the nitrogen cycling microbial community. The seasonal changes in abundances we observed also provided additional evidence of a dynamic relationship between microorganisms and plants, an important mechanism controlling ecosystem nitrogen cycling. The third study determined spatial and temporal interactions between AOA, AOB and NOB. These steps are related in both space and time, as the ammonia-oxidizers provide the necessary substrate for nitrite-oxidizers. Using a combination of spatial statistics and phylogenetic analysis, our data indicated seasonally varying patterns of niche differentiation between the two bacterial groups, Nitrospira and Nitrobacter in April, but more homogeneous patterns by August which may have been due to different strategies for adapting to changes in substrate concentrations resulting from competition with plants. We then asked a further question: was the microbial structure at sampling sites with high NS gene abundances fundamentally different from those with low NS gene abundances? Using a phylogenetic approach, the operational taxonomic unit composition of NS was analyzed. Community composition did not change over the first half of the season, but by the second half, the relative proportion of a particular OTU had increased significantly. This suggested an intraspecific competition within the NS and the possible importance of OTU 03 in nitrite oxidation at a specific period of time. Observed positive correlations between AOA and Nitrospira further suggested that in this unfertilized grassland plot, the nitrification process may be predominantly performed by these groups, but is restricted to a limited timeframe.
Ein Vergleich zwischen Barometrischer Prozessseparation (BaPS) und 15N-Verdünnungsmethode zur Bestimmung der Bruttonitrifikationsrate im Boden
(2010) Schwarz, Ulrich; Streck, Thilo
Besides the carbon cycle, the nitrogen cycle plays a central role in soil. A key process of this cycle is nitrification. In practice, nitrification is measured as gross or net nitrification. Net nitrification rates are measured by determining the net change in the nitrate or ammonium pool over a period of time. Net rates are difficult to interpret, because the net nitrification rate is the sum of nitrate producing and consuming processes. In contrast, gross nitrification quantifies the total production of nitrate via nitrification. There are two methods for measuring gross nitrification: the 15N-Pool dilution technique and Barometric Process Separation (BaPS). In the 15N-Pool dilution technique, nitrate en-riched with the heavier isotope 15N is added to soil, and the dilution of the 15N pool and the change in the nitrate pool are measured over time. The BaPS method measures changes in pressure and the oxygen- and carbon dioxide concentration of the atmosphere in a closed chamber. The gross nitrification rate can then be computed by a step-by-step solution of the gas balance equations. In the present study, 15N enriched nitrate was added to soil and then put into the BaPS-incubation chamber. By this procedure gross nitrification rates were measured simultaneously with both the 15N-Pool dilution technique and the BaPS method. The aim of the present study was to find out under which conditions the two methods yield similar results and under which conditions different results. In the latter case, the thesis aimed at elucidating the cause for the disagreement between both methods. For this purpose extensive research on two agricultural soils from North China and three soils from Southwest Germany was undertaken. The two methods were compared under the following conditions: 1) application of ammonium fertilizer, 2) addition of nitrification inhibitors, 3) varying soil wa-ter contents, and 4) different soil temperatures. Moreover, a new methodological approach was tested: the 13CO2-Pool dilution technique. Combining this method with the 15N-Pool dilu-tion technique and the Barometric Process Separation made it possible to exactly determine the pH and respiration coefficient in situ. Both techniques corresponded well in soil with pH<6. In soil with higher pH, both methods led to very different results. The reason is that pH has a strong impact on the calculation of the nitrification rate in the BaPS method. In nearly all experiments with neutral to alkaline soils, the BaPS technique yielded higher nitrification rates than the 15N-Pool dilution technique if pH was determined in 0.01 M CaCl2. With pH determined in water, there was good agreement or nitrification rates were too low. Fertilization with ammonium did not in-duce an increase of nitrification in a sandy soil with pH<6. A decrease in nitrification to less than 60% was achieved by the application of the nitrification inhibitor DCD. For both techniques a positive correlation between temperature and nitrification rates was found. There was no correlation between water filled pore space and nitrification rate.