Microbial communities display a variety of biogeographical patterns mainly driven by large-scale environmental gradients. Here, we analysed the spatial distribution of methane-oxidizing bacteria (MOB) and methane oxidation in a strongly fluctuating environment. We investigated whether the spatial variability of the MOB community can be explained by an environmental gradient and whether this changes with different plot sizes. We applied a pmoA-specific microarray to detect MOB, measured methane oxidation, methane emissions and soil properties. All variables were measured in a 10 × 10 m, 1 × 1 m and 20 × 20 cm plot and interpreted using a geostatistical approach. Methane oxidation as well as MOB displayed spatial patterns reflected in the underlying flooding gradient. Overlapping and contrasting spatial patterns for type I and type II MOB suggested different ecological life strategies. With smaller plot size, the environmental gradient could not explain the variability in the data and local factors became more important. In conclusion, environmental gradients can generally explain variability in microbial spatial patterns; however, we think that this does not contribute to a mechanistic explanation for microbial diversity because the relevant scales for microorganisms are much smaller than those normally measured.
Abstract Shallow aquatic systems exchange large amounts of carbon dioxide (CO 2 ) and methane (CH 4 ) with the atmosphere. The production and consumption of both gases is determined by the interplay between abiotic (such as oxygen availability) and biotic (such as community structure and trophic interactions) factors. Fish communities play a key role in driving carbon fluxes in benthic and pelagic habitats. Previous studies indicate that trophic interactions in the water column, as well as in the benthic zone can strongly affect aquatic CO 2 and CH 4 net emissions. However, the overall effect of fish on both pelagic and benthic processes remains largely unresolved, representing the main focus of our experimental study. We evaluated the effects of benthic and pelagic fish on zooplankton and macroinvertebrates; on CO 2 and CH 4 diffusion and ebullition, as well as on CH 4 production and oxidation, using a full‐factorial aquarium experiment. We compared five treatments: absence of fish (control); permanent presence of benthivorous fish (common carps, benthic) or zooplanktivorous fish (sticklebacks, pelagic); and intermittent presence of carps or sticklebacks. We found trophic and non‐trophic effects of fish on CO 2 and CH 4 emissions. Intermittent presence of benthivorous fish promoted a short‐term increase in CH 4 ebullition, probably due to the physical disturbance of the sediment. As CH 4 ebullition was the major contributor to the total greenhouse gas (GHG) emissions, incidental bioturbation by benthivorous fish was a key factor triggering total carbon emissions from our aquariums. Trophic effects impacted GHG dynamics in different ways in the water column and the sediment. Fish predation on zooplankton led to a top‐down trophic cascade effect on methane‐oxidising bacteria. This effect was, however, not strong enough as to substantially alter CH 4 diffusion rates. Top‐down trophic effects of zooplanktivorous and benthivorous fish on benthic macroinvertebrates, however, were more pronounced. Continuous fish predation reduced benthic macroinvertebrates biomass decreasing the oxygen penetration depth, which in turn strongly reduced water–atmosphere CO 2 fluxes while it increased CH 4 emission. Our work shows that fish can strongly impact GHG production and consumption processes as well as emission pathways, through trophic and non‐trophic effects. Furthermore, our findings suggest their impact on benthic organisms is an important factor regulating carbon (CO 2 and CH 4 ) emissions.
The rhizosphere of oxygen-releasing wetland plants provides a niche for oxygen-consuming microorganisms such as chemolithotrophic ammonia-oxidising bacteria.These bacteria are adapted to oxygen limitation with respect to their affinity for oxygen, ability to survive periods of anoxia, and immediate response to the appearance of oxygen.In this study the techniques of specific amplification of ammonia oxidiser 16S rDNA fragments by PCR, separation of mixed PCR samples by denaturing gradient gel electrophoresis (DGGE), and band identification by specific hybridisation with oligonucleotide probes were combined to allow for the comparison of the community composition of multiple samples over space and time.DGGE bands of interest were also excised for DNA isolation, reamplification, sequence determination and phylogenetic analysis.We compared monthly samples from both the root zone and the bare sediment of a shallow lake inhabited by the emergent macrophyte Glyceria maxima to determine the seasonal effects that the plant roots and the oxygen availability might have on the L-subgroup ammonia-oxidiser populations present.Similarly, five soil or sediment samples, varying in oxygen availability, from different locations in the Netherlands were compared.Although the presence of two previously defined Nitrosospira sequence clusters could be differentially detected in the samples examined, there was no evidence for a particular group which was specific to periodically anoxic environments.
Abstract Ammonium-induced stimulatory, inhibitory, and/or neutral effects on soil methane oxidation have been attributable to the ammonium concentration and mineral forms, confounded by other edaphic properties (e.g., pH, salinity), as well as the site-specific composition of the methanotrophic community. We hypothesize that this inconsistency may stem from the discrepancy in the cation adsorption capacity of the soil. We postulate that the effects of ammonium on the methanotrophic activity in soil are more accurately portrayed by relating methane uptake rates to the soluble ammonium (bioavailable), rather than the exchangeable (total) ammonium. To reduce adsorption (exchangeable) sites for ammonium in a paddy soil, two successive pre-incubation steps were introduced resulting in a 1000-fold soil dilution (soil enrichment), to be compared to a soil slurry (tenfold dilution) incubation. Ammonium was supplemented as NH 4 Cl at 0.5–4.75gL −1 after pre-incubation. While NH 4 Cl significantly stimulated the methanotrophic activity at all concentrations in the soil slurry incubation, methane uptake showed a dose-dependent effect in the soil enrichment. The trend in methane uptake could be explained by the soluble ammonium concentration, which was proportionate to the supplemented ammonium in the soil enrichment. In the soil slurry incubation, a fraction (36–63%) of the supplemented ammonium was determined to be adsorbed to the soil. Accordingly, Methylosarcina was found to predominate the methanotrophic community after the incubation, suggesting the relevance of this methanotroph at elevated ammonium levels (< 3.25gL −1 NH 4 Cl). Collectively, our results showed that the soluble, rather than the exchangeable ammonium concentration, is relevant when determining the effects of ammonium on methane oxidation, but this does not exclude other (a)biotic factors concurrently influencing methanotrophic activity.
The population dynamics of the chemolithoautotrophic nitrifiers Nitrosomonas europaea and Nitrobacter winogradskyi were studied in gnotobiotic microcosms fed with ammonium in response to the presence or absence of the emergent macrophyte Glyceria maxima and the heterotrophic denitrifying bacterium Pseudomonas chlororaphis. By subjecting the plants to different day lengths, the effect of possibly limiting factors (i.e. oxygen and ammonium) on the interactions between the nitrifiers and denitrifying bacterium could be analysed. The presence of the plant had no effect on the growth of nitrifiers suggesting that, in addition to radial oxygen loss from the roots, other non-plant sources of oxygen (e.g. diffusion from the water layer) were important for nitrification. Potential nitrifying activities were suppressed by G. maxima due to ammonium uptake by the plants. Elongation of the day length in combination with the presence of G. maxima led to an increase in the number of P. chlororaphis. The presence of P. chlororaphis suppressed the growth of N. winogradskyi, but the growth of N. europaea and the potential nitrifying activities were not significantly affected. Potential denitrifying activities were stimulated by the plant, but showed no correlations with nitrifier activities or numbers. Apparently ammonium, and not oxygen, was the limiting factor for nitrification in the root zone of G. maxima. However, when the plant did not deplete the ammonium pool, P. chlororaphis could repress the nitrifiers indicating the latter's poor competitive status with respect to oxygen when the presence of root exudates allows for heterotrophic oxygen consumption.
Methanotrophic communities were studied in several periodically water-saturated gleyic soils. When sampled, each soil had an oxic upper layer and consumed methane from the atmosphere (at 1.75 ppmv). In most gleyic soils the K(m(app)) values for methane were between 70 and 800 ppmv. These are higher than most values observed in dry upland soils, but lower than those measured in wetlands. Based on cultivation-independent retrieval of the pmoA-gene and quantification of partial pmoA gene sequences, type II (Alphaproteobacteria) methanotrophs of the genus Methylocystis spp. were abundant (> 10(7) pmoA target molecules per gram of dry soil). Type I (Gammaproteobacteria) methanotrophs related to the genera Methylobacter and Methylocaldum/Methylococcus were detected in some soils. Six pmoA sequence types not closely related to sequences from cultivated methanotrophs were detected as well, indicating that diverse uncultivated methanotrophs were present. Three Gleysols were incubated under different mixing ratios of (13)C-labelled methane to examine (13)C incorporation into phospholipid fatty acids (PLFAs). Phospholipid fatty acids typical of type II methanotrophs, 16:0 and 18:1omega7c, were labelled with (13)C in all soils after incubation under an atmosphere containing 30 ppmv of methane. Incubation under 500 ppmv of methane resulted in labelling of additional PLFAs besides 16:0 and 18:1omega7c, suggesting that the composition of the active methanotrophic community changed in response to increased methane supply. In two soils, 16:1 PLFAs typical of type I methanotrophs were strongly labelled after incubation under the high methane mixing ratio only. Type II methanotrophs are most likely responsible for atmospheric methane uptake in these soils, while type I methanotrophs become active when methane is produced in the soil.
The stoichiometry of prokaryotes (Bacteria and Archaea) can control benthic phosphorus (P) fluxes relative to carbon (C) and nitrogen (N) during organic matter remineralization. This paper presents the first experimental data on benthic microbial stoichiometry. We used X-ray microanalysis to determine C : N : P ratios of individual prokaryotes from C-limited Baltic Sea sediments incubated under oxic or anoxic conditions. At approximately 400:1, C : P ratios of prokaryotes from both oxic and anoxic incubations were higher than the Redfield ratio for marine organic matter (106:1), whereas prokaryotic C : N ratios (6.4:1) were close to the Redfield ratio. We conclude that high microbial C : P ratios contribute to the enhanced remineralization of P from organic matter relative to C and N observed in many low oxygen marine settings.
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