Summary Results from the pioneering research on the interactions between pH and denitrification in soil from the 1950s to the present are reviewed, the changing perceptions of this complex relationship are discussed, and the current status of the subject is assessed. Facets of this relationship that are analysed in detail include the direct or indirect influence of pH on overall denitrification rates in soils, changes in the composition of gaseous products that depend on pH, methods for measuring the process, the concept of an optimum pH for denitrification, and the adaptation of microbial denitrifying communities to acidic environments. The main conclusions to be drawn are as follows. Total gaseous emissions to the atmosphere (N2O, NO and N2) have repeatedly been shown to be less in acidic than in neutral or slightly alkaline soils. This may be attributable to smaller amounts of organic carbon and mineral nitrogen available to the denitrifying population under acid conditions rather than a direct effect of low pH on denitrification enzymes. Numerous laboratory and field studies have demonstrated that the ratio N2O:N2 is increased when the pH of soils is reduced. The relation between soil pH and potential denitrification as determined by various incubation methods remains unclear, results being influenced both by original conditions in soil samples and by unknown changes during incubation. The concept of an optimum pH for denitrification has been frequently proposed, but such a term has little or no meaning without reference to specific attributes of the process.

There is ample evidence that microbial processes can exhibit large variations in activity on a field scale. However, very little is known about the spatial distribution of the microbial communities mediating these processes. Here we used geostatistical modelling to explore spatial patterns of size and activity of the denitrifying community, a functional guild involved in N-cycling, in a grassland field subjected to different cattle grazing regimes. We observed a non-random distribution pattern of the size of the denitrifier community estimated by quantification of the denitrification genes copy numbers with a macro-scale spatial dependence (6-16 m) and mapped the distribution of this functional guild in the field. The spatial patterns of soil properties, which were strongly affected by presence of cattle, imposed significant control on potential denitrification activity, potential N(2)O production and relative abundance of some denitrification genes but not on the size of the denitrifier community. Absolute abundance of most denitrification genes was not correlated with the distribution patterns of potential denitrification activity or potential N(2)O production. However, the relative abundance of bacteria possessing the nosZ gene encoding the N(2)O reductase in the total bacterial community was a strong predictor of the N(2)O/(N(2) + N(2)O) ratio, which provides evidence for a relationship between bacterial community composition based on the relative abundance of denitrifiers in the total bacterial community and ecosystem processes. More generally, the presented geostatistical approach allows integrated mapping of microbial communities, and hence can facilitate our understanding of relationships between the ecology of microbial communities and microbial processes along environmental gradients.

The objective of this study was to investigate how changes in soil pH affect the N 2 O and N 2 emissions, denitrification activity, and size of a denitrifier community. We established a field experiment, situated in a grassland area, which consisted of three treatments which were repeatedly amended with a KOH solution (alkaline soil), an H 2 SO 4 solution (acidic soil), or water (natural pH soil) over 10 months. At the site, we determined field N 2 O and N 2 emissions using the 15 N gas flux method and collected soil samples for the measurement of potential denitrification activity and quantification of the size of the denitrifying community by quantitative PCR of the narG, napA, nirS, nirK, and nosZ denitrification genes. Overall, our results indicate that soil pH is of importance in determining the nature of denitrification end products. Thus, we found that the N 2 O/(N 2 O ؉ N 2 ) ratio increased with decreasing pH due to changes in the total denitrification activity, while no changes in N 2 O production were observed. Denitrification activity and N 2 O emissions measured under laboratory conditions were correlated with N fluxes in situ and therefore reflected treatment differences in the field. The size of the denitrifying community was uncoupled from in situ N fluxes, but potential denitrification was correlated with the count of NirS denitrifiers. Significant relationships were observed between nirS, napA, and narG gene copy numbers and the N 2 O/(N 2 O ؉ N 2 ) ratio, which are difficult to explain. However, this highlights the need for further studies combining analysis of denitrifier ecology and quantification of denitrification end products for a comprehensive understanding of the regulation of N fluxes by denitrification.Denitrification is the microbial reduction of NO 3 Ϫ via NO 2 Ϫ to gaseous NO, N 2 O, and N 2 , which are then lost into the atmosphere (36). It therefore results in considerable loss of nitrogen, one of the most limiting nutrients for crop production in agriculture (20). Denitrification is also of environmental concern since, together with nitrification, it is the main biological process responsible for N 2 O emissions (7). N 2 O is a potent greenhouse gas which has a global warming potential about 320 times greater than that of CO 2 and has a lifetime of approximately 120 years (32). In the stratosphere, N 2 O can also react with O 2 to produce NO, which induces the destruction of stratospheric ozone (8). N 2 O can be released into the atmosphere by incomplete denitrification due to the effect of environmental conditions on the regulation of the different denitrification reductases (14, 41, 51), but it has recently been suggested that it could also be due to lack of nitrous oxide reductase in some denitrifiers (19,41). Since N 2 O is an intermediate in the denitrification pathway, both the amount of N 2 O produced and the N 2 O/(N 2 O ϩ N 2 ) ratio are important in understanding and predicting N 2 O fluxes from soils. The main environmental factors known to influence the N 2 O/(N 2 O...

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Whether bacteria display spatial patterns of distribution and at which level of taxonomic organization such patterns can be observed are central questions in microbial ecology. Here we investigated how the total and relative abundances of eight bacterial taxa at the phylum or class level were spatially distributed in a pasture by using quantitative PCR and geostatistical modelling. The distributions of the relative abundance of most taxa varied by a factor of 2.5-6.5 and displayed strong spatial patterns at the field scale. These spatial patterns were taxon-specific and correlated to soil properties, which indicates that members of a bacterial clade defined at high taxonomical levels shared specific ecological traits in the pasture. Ecologically meaningful assemblages of bacteria at the phylum or class level in the environment provides evidence that deep branching patterns of the 16S rRNA bacterial tree are actually mirrored in nature.

In the present study, we tested the hypothesis that animal treading associated with a high input of organic matter would favour methanogenesis in soils used as overwintering pasture. Hence, methane emissions and methanogen populations were examined at sections with different degree of cattle impact in a Farm in South Bohemia, Czech Republic. In spring, methane emission positively corresponded to the gradient of animal impact. Applying phospholipid etherlipid analysis, the highest archaeal biomass was found in section severe impact (SI), followed by moderate impact (MI) and no impact. The same trend was observed for the methanogens as showed by real-time quantitative PCR analyses of methyl coenzyme M reductase (mcrA) genes. The detection of monounsaturated isoprenoid side chain hydrocarbons (i20:1) indicated the presence of acetoclastic methanogens in the cattle-impacted sites. This result was corroborated by the phylogenetic analysis of mcrA gene sequences obtained from section SI, which showed that 33% of the analysed clones belonged to the genus Methanosarcina. The majority of the sequenced clones (41%) showed close affiliations with uncultured rumen archaeons. This leads to the assumption that a substantial part of the methanogenic community in plot SI derived from the grazing cattle itself. Compared to the spring sampling, in autumn, a significant reduction in archaeal biomass and number of copies of mcrA genes was observed mainly for section MI. It can be concluded that after 5 months without cattle impact, the severely impact section maintained its methane production potential, whereas the methane production potential under moderate impact returned to background values.

Soil microbial properties were studied from localities on a transect along the Yenisei River, Central Siberia. The 1000 km‐long transect, from 56°N to 68°N, passed through tundra, taiga and pine forest characteristic of Northern Russia. Soil microbial properties were characterized by dehydrogenase activity, microbial biomass, composition of microbial community (PLFAs), respiration rates, denitrification and N mineralization rates. Relationships between vegetation, latitude, soil quality (pH, texture), soil organic carbon (SOC) and the microbial properties were examined using multivariate analysis. In addition, the temperature responses of microbial growth (net growth rate) and activity (soil respiration rate) were tested by laboratory experiments. The major conclusions of the study are as follows: 1. Multivariate analysis of the data revealed significant differences in microbial activity. SOC clay content was positively related to clay content. Soil texture and SOC exhibited the dominant effect on soil microbial parameters, while the vegetation and climatic effects (expressed as a function of latitude) were weaker but still significant. The effect of vegetation cover is linked to SOC quality, which can control soil microbial activity. 2. When compared to fine‐textured soils, coarse‐textured soils have (i) proportionally more SOC bound in microbial biomass, which might result in higher susceptibility of SOC transformation to fluctuation of environmental factors, and (ii) low mineralization potential, but with a substantial part of the consumed C being transformed to microbial products. 3. The soil microbial community from the northernmost study region located within the permafrost zone appears to be adapted to cold conditions. As a result, microbial net growth rate became negative when temperature rose above 5 °C and C mineralization then exceeded C accumulation.

In the non-heterocyst, marine cyanobacterium Trichodesmium nitrogen fixation is confined to the photoperiod and occurs coevally with oxygenic photosynthesis although nitrogenase is irreversibly inactivated by oxygen. In previous studies it was found that regulation of photosynthesis for nitrogen fixation involves Mehler reaction and various activity states with reversible coupling of photosynthetic components. We now investigated these activity states in more detail. Spectrally resolved fluorescence kinetic measurements of single cells revealed that they were related to alternate uncoupling and coupling of phycobilisomes from and to the photosystems, changing the effective cross-section of PSII. Therefore, we isolated and purified the phycobiliproteins of Trichodesmium via ion exchange chromatography and recorded their UV/VIS absorption, fluorescence excitation and fluorescence emission spectra. After describing these spectra by mathematical equations via the Gauss-Peak-Spectra method, we used them to deconvolute the in vivo fluorescence spectra of Trichodesmium cells. This revealed that the contribution of different parts of the phycobilisome antenna to fluorescence quenching changed during the daily activity cycle, and that individual phycobiliproteins can be reversibly coupled to the photosystems, while the expression levels of these proteins did not change much during the daily activity cycle. Thus we propose that variable phycobilisome coupling plays a key role in the regulation of photosynthesis for nitrogen fixation in Trichodesmium.