Rebekka R. E. Artz

A stable bioluminescent construct of Escherichia coli O157:H7 for hazard assessments of long-term survival in the environment.

ABSTRACT A chromosomally lux-marked (Tn5 luxCDABE) strain of nontoxigenic Escherichia coli O157:H7 was constructed by transposon mutagenesis and shown to have retained the O157, H7, and intimin phenotypes. The survival characteristics of this strain in the experiments performed (soil at −5, −100, and −1,500 kPa matric potential and artificial groundwater) were indistinguishable from the wild-type strain. Evaluation of potential luminescence was found to be a rapid, cheap, and quantitative measure of viable E. coli O157:H7 Tn5 luxCDABE populations in environmental samples. In the survival studies, bioluminescence of the starved populations of E. coli O157:H7 Tn5 luxCDABE could be reactivated to the original levels of light emission, suggesting that these populations remain viable and potentially infective to humans. The attributes of the construct offer a cheap and low-risk substitute to the use of verocytotoxin-producing E. coli O157:H7 in long-term survival studies.

Changes in fungal community composition in response to vegetational succession during the natural regeneration of cutover peatlands

Despite the importance of peatlands as a major store of sequestered carbon and the role of fungi in releasing sequestered C, we know little about the community structure of fungi in peatlands. We investigated these across a gradient of naturally regenerating peatland vegetation using denaturing gradient gel electrophoresis (DGGE) and clone libraries of fragments of the fungal rRNA internal transcribed spacer (ITS) region. Significant changes in the fungal community structure of peat samples at different stages of regeneration were observed, which relate to the composition of the vegetation recolonizing these sites. Cloning and sequence analysis also demonstrated a potential shift in the relative abundance of the main fungal phyla. Some of the clones identified to genus level were highly related to fungi known to play a role in the degradation of plant litter or wood in similar ecosystems and/or form mycorrhizal associations. In addition, several fungal isolates highly related to peat clones were obtained, and their enzymic capacity to degrade structural plant tissues was assessed. Together, these results suggest that the fungal community composition of peat may be an important indicator of the status of regeneration and potential carbon sequestration of cutover peatlands.

Summer drought decreases soil fungal diversity and associated phenol oxidase activity in upland Calluna heathland soil

Natural moisture limitation during summer drought can constitute a stress for microbial communities in soil. Given globally predicted increases in drought frequency, there is an urgent need for a greater understanding of the effects of drought events on soil microbial processes. Using a long-term field-scale drought manipulation experiment at Clocaenog, Wales, UK, we analysed fungal community dynamics, using internal transcribed spacer-denaturing gradient gel electrophoresis (DGGE), over a 1-year period in the 6th year of drought manipulation. Ambient seasonality was found to be the dominant factor driving variation in fungal community dynamics. The summer drought manipulation resulted in a significant decline in the abundance of dominant fungal species, both independently of, and in interaction with, this seasonal variation. Furthermore, soil moisture was significantly correlated with the changes in fungal diversity over the drought manipulation period. While the relationship between species diversity and functional diversity remains equivocal, phenol oxidase activity was decreased by the summer drought conditions and there was a significant correlation with the decline of DGGE band richness among the most dominant fungal species during the drought season. Climatically driven events such as droughts may have significant implications for fungal community diversity and therefore, have the potential to interfere with crucial ecosystem processes, such as organic matter decomposition.

Hydrogenosomes in the anaerobic fungus Neocallimastix frontalis have a double membrane but lack an associated organelle genome

The presence of hydrogenosomes in phylogenetically distinct anaerobic eukaryotes implies that they have been acquired independently, and previously reported differences in ultrastructure among taxa have suggested that some hydrogenosomes have different origins. Of particular interest are reports that Neocallimastix frontalis hydrogenosomes resemble microbodies in possessing a single membrane, in contrast to those in ciliates and trichomonads which have two and thus resemble mitochondria. In this investigation we have clearly demonstrated that N. frontalis hydrogenosomes possess two, rather than one, closely apposed membranes and in some preparations cristae‐like structures were observed. These observations have led us to reject the microbody hypothesis and provide some indirect support for a possible mitochondrion origin as proposed for other hydrogenosomes. N. frontalis hydrogenosomes were shown to lack an associated genome as previously demonstrated for trichomonad hydrogenosomes. This might be explained by assuming that a mitochondrial genome encoding proteins for aerobic function is no longer necessary for either organelle.

Long-term nitrogen deposition increases phosphorus limitation of bryophytes in an ombrotrophic bog

Here we investigate the effect of 4 years simulated atmospheric deposition of ammonium (NH4) and nitrate (NO3), applied alone or in combination with phosphorus and potassium (PK), on the surface phosphatase activities and nutrient acquisition behaviour of two species of moss (Sphagnumcapillifolium and Hypnum jutlandicum) from an ombrotrophic peatland. Phosphatase activity was significantly enhanced by both the NH4 and NO3 treatments, particularly for Sphagnum, but the activity decreased when exposed to additions of PK. Regression analysis revealed that phosphatase activity on Sphagnum was positively related with tissue N and negatively related to tissue P concentrations. For Hypnum, a negative relationship between shoot P concentration and phosphatase activity was observed. Using a 32P tracer, mosses removed from plots receiving PK in combination with NH4 maintained their affinity for increased phosphorus uptake. These findings suggest that enhanced nutrient supply, even at modest doses, significantly alter the nutrient recycling behaviour of bryophytes.

Interactions among fungal community structure, litter decomposition and depth of water table in a cutover peatland

Peatlands are important reservoirs of carbon (C) but our understanding of C cycling on cutover peatlands is limited. We investigated the decomposition over 18 months of five types of plant litter (Calluna vulgaris, Eriophorum angustifolium, Eriophorum vaginatum, Picea sitchensis and Sphagnum auriculatum) at a cutover peatland in Scotland, at three water tables. We measured changes in C, nitrogen (N) and phosphorus (P) in the litter and used denaturing gradient gel electrophoresis to investigate changes in fungal community composition. The C content of S. auriculatum litter did not change throughout the incubation period whereas vascular plant litters lost 30-40% of their initial C. There were no differences in C losses between low and medium water tables, but losses were always significantly less at the high water table. Most litters accumulated N and E. angustifolium accumulated significant quantities of P. C, N and P were significant explanatory variables in determining changes in fungal community composition but explained <25% of the variation. Litter type was always a stronger factor than water table in determining either fungal community composition or turnover of C, N and P in litter. The results have implications for the ways restoration programmes and global climate change may impact upon nutrient cycling in cutover peatlands.

Effect of Nematodes on Rhizosphere Colonization by Seed-Applied Bacteria

ABSTRACT There is much interest in the use of seed-applied bacteria for biocontrol and biofertilization, and several commercial products are available. However, many attempts to use this strategy fail because the seed-applied bacteria do not colonize the rhizosphere. Mechanisms of rhizosphere colonization may involve active bacterial movement or passive transport by percolating water or plant roots. Transport by other soil biota is likely to occur, but this area has not been well studied. We hypothesized that interactions with soil nematodes may enhance colonization. To test this hypothesis, a series of microcosm experiments was carried out using two contrasting soils maintained under well-defined physical conditions where transport by mass water flow could not occur. Seed-applied Pseudomonas fluorescens SBW25 was capable of rhizosphere colonization at matric potentials of −10 and −40 kPa in soil without nematodes, but colonization levels were substantially increased by the presence of nematodes. Our results suggest that nematodes can have an important role in rhizosphere colonization by bacteria in soil.

Long term repeated prescribed burning increases evenness in the basidiomycete laccase gene pool in forest soils

Repeated prescribed burning alters the biologically labile fraction of nutrients and carbon of soil organic matter (SOM). Using a long-term (30 years) repeated burning experiment where burning has been carried out at a 2- or 4-year frequency, we analysed the effect of prescribed burning on gross potential C turnover rates and phenol oxidase activity in relation to shifts in SOM composition as observed using Fourier-transform infrared spectroscopy. In tandem, we assessed the genetic diversity of basidiomycete laccases. While the overall effect of burning was a decline in phenol oxidase activity, Shannon diversity and evenness of laccases was significantly higher in burned sites. Co-correspondence analysis of SOM composition and laccase operational taxonomic unit frequency data also suggested a strong correlation. While this correlation could indicate that the observed increase in laccase genetic diversity due to burning is due to increased resource diversity, a temporal replacement of the most abundant members of the assembly by an otherwise dormant pool of fungi cannot be excluded. As such, our results fit the intermediate disturbance hypothesis. Effects were stronger in plots burned in 2-year rotations, suggesting that the 4-year burn frequency may be a more sustainable practice to ensure the long-term stability of C cycling in such ecosystems.

Microbial Community Structure and Carbon Substrate use in Northern Peatlands

The net C sink function of many northern peatlands is a product of an imperfect balance between net primary production and net respiration. This review summarizes our current knowledge of the microbial pathways of carbon flow in such ecosystems, the key players in the aerobic and anaerobic decomposition routes, and the drivers of microbial community structure and substrate use. The review identifies several areas in need of future research, notably on the key players and their activities in the periodically oxic and the permanently anoxic zones of peatlands.

Potential for using remote sensing to estimate carbon fluxes across northern peatlands – A review

Peatlands store large amounts of terrestrial carbon and any changes to their carbon balance could cause large changes in the greenhouse gas (GHG) balance of the Earths atmosphere. There is still much uncertainty about how the GHG dynamics of peatlands are affected by climate and land use change. Current field-based methods of estimating annual carbon exchange between peatlands and the atmosphere include flux chambers and eddy covariance towers. However, remote sensing has several advantages over these traditional approaches in terms of cost, spatial coverage and accessibility to remote locations. In this paper, we outline the basic principles of using remote sensing to estimate ecosystem carbon fluxes and explain the range of satellite data available for such estimations, considering the indices and models developed to make use of the data. Past studies, which have used remote sensing data in comparison with ground-based calculations of carbon fluxes over Northern peatland landscapes, are discussed, as well as the challenges of working with remote sensing on peatlands. Finally, we suggest areas in need of future work on this topic. We conclude that the application of remote sensing to models of carbon fluxes is a viable research method over Northern peatlands but further work is needed to develop more comprehensive carbon cycle models and to improve the long-term reliability of models, particularly on peatland sites undergoing restoration.