Hay strewing, brush harvesting of seed and soil disturbance as tools for the enhancement of botanical diversity in grasslands

Since the middle of the last century agricultural intensification within Europe has led to a drastic decline in the extent of botanically diverse grasslands. Whilst measures to enhance the diversity of agriculturally-improved grasslands are in place, success has often been limited. One of the primary factors limiting success is the paucity of sources of propagules of desirable species in the surrounding landscape. The restoration of two contrasting grassland types lowland hay meadow and chalk grassland) was examined using a replicated block experiment to assess the effectiveness of two methods of seed application (hay strewing and brush harvesting) and two methods of pre-treatment disturbance (power harrowing and turf stripping). The resulting changes in botanical composition were monitored for 4 years. Seed addition by both methods resulted in significant temporal trends in plant species composition and increases in plant species richness, which were further enhanced by disturbance. Power harrowing increased the effectiveness of the seed addition treatments at the lowland hay meadow site. At the chalk grassland site a more severe disturbance created by turf stripping was used and shown to be preferable. Whilst both hay strewing and brush harvesting increased plant species richness, hay strewing was more effective at creating a sward similar to that of the donor site. Soil disturbance and seed application rate at the recipient site and timing of the hay cut at the donor site are all factors to be considered prior to the commencement of restoration management. (c) 2006 Elsevier Ltd. All rights reserved.

Plant species diversity, plant biomass and responses of the soil community on abandoned land across Europe: idiosyncracy or above-belowground time lags

The relationship between plant species diversity, productivity and the development of the soil community during early secondary succession on former arable land across Europe is investigated. The enhancement of biomass production due to the increase in initial plant species diversity and the consequent stimulation of soil microbial biomass and abundance of soil invertebrates are examined.

Plant-soil interactions and grassland diversity restoration

Restoration schemes aimed at enhancing plant species diversity of improved agricultural grassland have been a key feature of agri-environmental policy since the mid 1980s. Allied to this has been much research aimed at providing policy makers with guidelines on how best to manage grassland to restore botanical diversity. This research includes long-term studies of the consequences for grassland diversity of management techniques such as different hay cut dates, fertiliser additions, seed introductions and grazing regimes. Studies have also explored the role of introductions of Rhinanthus minor into species-poor swards to debilitate competitive grasses. While these studies have been successful in identifying some management features that control plant species diversity in agricultural grassland, they have taken a largely aboveground perspective on plant community dynamics.

Biodegradation of the herbicide mecoprop-p with soil depth and its relationship with class III tfdA genes

Mecoprop-p [(R)-2-(4-chloro-2-methylphenoxy) propanoic acid) is widely used in agriculture and poses an environmental concern because of its susceptibility to leach from soil to water. We investigated the effect of soil depth on mecoprop-p biodegradation and its relationship with the number and diversity of tfdA related genes, which are the most widely known genes involved in degradation of the phenoxyalkanoic acid group of herbicides by bacteria. Mecoprop-p half-life (DT50) was approximately 12 days in soil sampled from <30 cm depth, and increased progressively with soil depth, reaching over 84 days at 70–80 cm. In sub-soil there was a lag period of between 23 and 34 days prior to a phase of rapid degradation. No lag phase occurred in top-soil samples prior to the onset of degradation. The maximum degradation rate was the same in top-soil and sub-soil samples. Although diverse tfdAα and tfdA genes were present prior to mecoprop-p degradation, real time PCR revealed that degradation was associated with proliferation of tfdA genes. The number of tfdA genes and the most probable number of mecoprop-p degrading organisms in soil prior to mecoprop-p addition were below the limit of quantification and detection respectively. Melting curves from the real time PCR analysis showed that prior to mecoprop-p degradation both class I and class III tfdA genes were present in top- and sub-soil samples. However at all soil depths only tfdA class III genes proliferated during degradation. Denaturing gradient gel electrophoresis confirmed that class III tfdA genes were associated with mecoprop-p degradation. Degradation was not associated with the induction of novel tfdA genes in top- or sub-soil samples, and there were no apparent differences in tfdA gene diversity with soil depth prior to or following degradation.

Microbial and biochemical soil quality indicators and their potential for differentiating areas under contrasting agricultural management regimes

The aim of this study was to examine interrelationships between functional biochemical and microbial indicators of soil quality, and their suitability to differentiate areas under contrasting agricultural management regimes. The study included five 0.8 ha areas on a sandy-loam soil which had received contrasting fertility and cropping regimes over a 5 year period. These were organically managed vegetable, vegetable -cereal and arable rotations, an organically managed grass clover ley, and a conventional cereal rotation. The organic areas had been converted from conventional cereal production 5 years prior to the start of the study. All of the biochemical analyses, including light fraction organic matter (LFOM) C and N, labile organic N (LON), dissolved organic N and water-soluble carbohydrates showed significant differences between the areas, although the nature of the relationships between the areas varied between the different parameters, and were not related to differences in total soil organic matter content. The clearest differences were seen in LFOM C and N and LON, which were higher in the organic arable area relative to the other areas. In the case of the biological parameters, there were differences between the areas for biomass-N, ATP, chitin content, and the ratios of ATP: biomass and basal respiration: biomass. For these parameters, the precise relationships between the areas varied. However, relative to the conventionally managed area, areas under organic management generally had lower biomass-N and higher ATP contents. Arbuscular mycorrhizal fungus colonization potential was extremely low in the conventional area relative to the organic areas. Further, metabolic diversity and microbial community level physiological profiles, determined by analysis of microbial community metabolism using Biolog GN plates and the activities of eight key nutrient cycling enzymes, grouped the organic areas together, but separated them from the conventional area. We conclude that microbial parameters are more effective and consistent indicators of management induced changes to soil quality than biochemical parameters, and that a variety of biochemical and microbial analyses should be used when considering the impact of management on soil quality. (C) 2004 Elsevier Ltd. All rights reserved.

Indicators of nitrate in wetland surface and soil-waters: interactions of vegetation and environmental factors

This paper describes a new bio-indicator method for assessing wetland ecosystem health: as such, the study is particularly relevant to current legislation such as the EU Water Framework Directive, which provides a baseline of the current status Of Surface waters. Seven wetland sites were monitored across northern Britain, with model construction data for predicting, eco-hydroloplical relationships collected from five sites during 1999, Two new sites and one repeat site were monitored during 2000 to provide model test data. The main growing season for the vegetation, and hence the sampling period, was May-August during both years. Seasonal mean concentrations of nitrate (NO3-) in surface and soil water samples during 1999 ranged from 0.01 to 14.07 mg N 1(-1), with a mean value of 1.01 mg N 1(-1). During 2000, concentrations ranged from trace level (<0.01 m- N 1(-1)) to 9.43 mg N 1(-1), with a mean of 2.73 mg N 1(.)(-1) Surface and soil-water nitrate concentrations did not influence plant species composition significantly across representative tall herb fen and mire communities. Predictive relationships were found between nitrate concentrations and structural characteristics of the wetland vegetation, and a model was developed which predicted nitrate concentrations from measures of plant diversity, canopy structure and density of reproductive structures. Two further models, which predicted stem density and density of reproductive structures respectively, utilised nitrate concentration as one of the independent predictor variables. Where appropriate, the models were tested using data collected during 2000. This approach is complementary to species-based monitoring, representing a useful and simple too] to assess ecological status in target wetland systems and has potential for bio-indication purposes.

Hydrological controls on soil redox dynamics in a peat-based, restored wetland

Increasing areas of altered wetland are being restored by re-flooding the soil. Evidence in the literature indicates that this practice can induce the redox-mediated release of soil nutrients, thereby increasing the risk of diffuse water pollution. However, for the sake of improving wedand management decisions, there is a need for more detailed studies of the underlying relationship between the hydrological and redox dynamics that explain this risk; this is particularly the case in agricultural peatlands that are commonly targeted for the creation of lowland wet grassland. A 12-month field study was conducted to evaluate the relationship between hydrological fluctuations and soil redox potential (Eh) in a nutrient-rich peat field (32 g N kg(-1) and 1100 mg P kg(-1) in the surface 0-30 cm soil) that had been restored as lowland wet grassland from intensive arable production. Field tensiometers were installed at the 30-, 60- and 90-cm soil depths, and Pt electrodes at the 10-, 30-, 60- and 90-cm depths, for daily logging of soil water tension and Eh, respectively. The values for soil water tension displayed a strong negative relationship (P < 0.001) with monthly dip well observations of water table height. Calculations of soil water potential from the logged tension values were used, therefore, to provide a detailed profile of field water level and, together with precipitation data, explained some of the variation in Eh. For example, during the summer, alternating periods of aerobism (Eh > 330 mV) in the surface, 0-10 cm layer of peat coincided with intense precipitation events. Redox potential throughout the 30-100 cm profile also fluctuated seasonally; indeed, at all depths Eh displayed a strong, negative relationship (P < 0.001) with water table height over the 12-month study period. However, Eh throughout the 30-100 cm profile remained relatively low (< 230 mV), indicating permanently reduced conditions that are associated with denitrification and reductive dissolution of Fe-bound P. The implications of these processes in the N- and P-rich peat for wetland plant diversity and water quality are discussed. (c) 2006 Elsevier B.V. All rights reserved.

Long-term response of the nematode community to elevated atmospheric CO2 in a temperate dry grassland soil

Long-term effects of the elevated atmospheric CO2 on biosphere have been in focus of research since the last few decades. In this experiment undisturbed soil monoliths of loess grassland were exposed to an elevated CO2 environment (two-times the ambient CO2 level) for a period of six years with the aid of the open top chamber method. Control without a chamber and CO2 elevation was applied as well. Elevated CO2 level had very little impact oil soil food web. It did not influence either root and microbial biomass or microbial and nematode community structure. The only significant response was that density of the bacterial feeder genus Heterocephalobus increased in the chamber with elevated CO2 concentration. Application of the open top chambers initiated more changes on nematodes than the elevated CO2 level. Open top chamber (OTC) method decreased nematode density (total and plant feeder as well) to less than half of the original level. Negative effect was found on the genus level in the case of fungal feeder Aphelenchoides, plant feeder Helicotylenchus and Paratylenchus. It is very likely that the significantly lower belowground root biomass and partly its decreased quality reflected by the increased C/N ratio are the main responsible factors for the lower density of the plant feeder nematodes in the plots of chambers. According to diversity profiles, MI and MI(2-15) parameters, nematode communities in the open top chambers (both on ambient and elevated CO2 level) seem to be more structured than those under normal circumstances six years after start of the experiment.

The value of sown grass margins for enhancing soil macrofaunal biodiversity in arable systems

The presence of a grass strip was found to be beneficial to soil macrofauna, increasing the species densities and abundances of earthworms, woodlice and staphylinid beetles. The biodiversity of the three main feeding groups - predators, soil ingesters and litter consumers - was also significantly higher in the grass strips than in the field edges without strips, indicating that establishment of grassy margins in arable fields may enhance ecosystem services such as soil fertility and pest control. The grass strip habitat contained a large number of species of soil macrofauna, being second only to hedgerow habitat, with 10% of the total species list for the farm found only within the margins. Of the rare species recorded on the farm, five of the nine were from the grass strips, four of which were found only there. This study shows that establishing grassy strips in the margins of arable fields increases the biodiversity of the soil macrofauna, both within fields (alpha diversity) and across the farm (beta diversity). (C) 2008 Elsevier B.V. All rights reserved.

The value of arable field margins for enhancing soil macrofaunal biodiversity in agroecosystems

The presence of a grass strip was found to be beneficial to soil macrofauna, increasing the species densities and abundances of earthworms, woodlice and staphylinid beetles. The biodiversity of the three main feeding groups – predators, soil ingesters and litter consumers – was also significantly higher in the grass strips than in the field edges without strips, indicating that establishment of grassy margins in arable fields may enhance ecosystem services such as soil fertility and pest control. The grass strip habitat contained a large number of species of soil macrofauna, being second only to hedgerow habitat, with 10% of the total species list for the farm found only within the margins. Of the rare species recorded on the farm, five of the nine were from the grass strips, four of which were found only there. This study shows that establishing grassy strips in the margins of arable fields increases the biodiversity of the soil macrofauna, both within fields (alpha diversity) and across the farm (beta diversity).

Can arable field margins be managed to enhance their biodiversity, conservation and functional value for soil macrofauna?

1. The establishment of grassy strips at the margins of arable fields is an agri-environment scheme that aims to provide resources for native flora and fauna and thus increase farmland biodiversity. These margins can be managed to target certain groups, such as farmland birds and pollinators, but the impact of such management on the soil fauna has been poorly studied. This study assessed the effect of seed mix and management on the biodiversity, conservation and functional value of field margins for soil macrofauna. 2. Experimental margin plots were established in 2001 in a winter wheat field in Cambridgeshire, UK, using a factorial design of three seed mixes and three management practices [spring cut, herbicide application and soil disturbance (scarification)]. In spring and autumn 2005, soil cores taken from the margin plots and the crop were hand-sorted for soil macrofauna. The Lumbricidae, Isopoda, Chilopoda, Diplopoda, Carabidae and Staphylinidae were identified to species and classified according to feeding type. 3. Diversity in the field margins was generally higher than in the crop, with the Lumbricidae, Isopoda and Coleoptera having significantly more species and/or higher abundances in the margins. Within the margins, management had a significant effect on the soil macrofauna, with scarified plots containing lower abundances and fewer species of Isopods. The species composition of the scarified plots was similar to that of the crop. 4. Scarification also reduced soil- and litter-feeder abundances and predator species densities, although populations appeared to recover by the autumn, probably as a result of dispersal from neighbouring plots and boundary features. The implications of the responses of these feeding groups for ecosystem services are discussed. 5. Synthesis and applications. This study shows that the management of agri-environment schemes can significantly influence their value for soil macrofauna. In order to encourage the litter-dwelling invertebrates that tend to be missing from arable systems, agri-environment schemes should aim to minimize soil cultivation and develop a substantial surface litter layer. However, this may conflict with other aims of these schemes, such as enhancing floristic and pollinator diversity.

Genomic and genetic analyses of diversity and plant interactions of Pseudomonas fluorescens

Background: Pseudomonas fluorescens are common soil bacteria that can improve plant health through nutrient cycling, pathogen antagonism and induction of plant defenses. The genome sequences of strains SBW25 and Pf0-1 were determined and compared to each other and with P. fluorescens Pf-5. A functional genomic in vivo expression technology (IVET) screen provided insight into genes used by P. fluorescens in its natural environment and an improved understanding of the ecological significance of diversity within this species. Results: Comparisons of three P. fluorescens genomes (SBW25, Pf0-1, Pf-5) revealed considerable divergence: 61% of genes are shared, the majority located near the replication origin. Phylogenetic and average amino acid identity analyses showed a low overall relationship. A functional screen of SBW25 defined 125 plant-induced genes including a range of functions specific to the plant environment. Orthologues of 83 of these exist in Pf0-1 and Pf-5, with 73 shared by both strains. The P. fluorescens genomes carry numerous complex repetitive DNA sequences, some resembling Miniature Inverted-repeat Transposable Elements (MITEs). In SBW25, repeat density and distribution revealed 'repeat deserts' lacking repeats, covering approximately 40% of the genome. Conclusions: P. fluorescens genomes are highly diverse. Strain-specific regions around the replication terminus suggest genome compartmentalization. The genomic heterogeneity among the three strains is reminiscent of a species complex rather than a single species. That 42% of plant-inducible genes were not shared by all strains reinforces this conclusion and shows that ecological success requires specialized and core functions. The diversity also indicates the significant size of genetic information within the Pseudomonas pan genome.

Belowground interactive effects of elevated CO2, plant diversity and earthworms in grassland microcosms

The potential interactive effects of future atmospheric CO2 concentrations and plant diversity loss on the functioning of belowground systems are still poorly understood. Using a microcosm greenhouse approach with assembled grassland plant communities of different diversity (1, 4 and 8 species), we explored the interactive effects between plant species richness and elevated CO2 (ambient and + 200 p.p.m.v. CO2) on earthworms and microbial biomass. We hypothesised that the beneficial effect of increasing plant species richness on earthworm performance and microbial biomass will be modified by elevated CO2 through impacts on belowground organic matter inputs, soil water availability and nitrogen availability. We found higher earthworm biomass in eight species mixtures under elevated CO2, and higher microbial biomass under elevated CO2 in four and eight species mixtures if earthworms were present. The results suggest that plant driven changes in belowground organic matter inputs, soil water availability and nitrogen availability explain the interactive effects of CO2 and plant diversity on the belowground compartment. The interacting mechanisms by which elevated CO2 modified the impact of plant diversity on earthworms and microorganisms are discussed.

Evaluation of the environmental specificity of fluorescence in situ hybridization (FISH) using fluorescence-activated cell sorting (FACS) of probe (PSE1284)-positive cells extracted from rhizosphere soil

We explicitly tested for the first time the ‘environmental specificity’ of traditional 16S rRNAtargeted fluorescence in situ hybridization (FISH) through comparison of the bacterial diversity actually targeted in the environment with the diversity that should be exactly targeted (i.e. without mismatches) according to in silico analysis. To do this, we exploited advances in modern Flow Cytometry that enabled improved detection and therefore sorting of sub-micron-sized particles and used probe PSE1284 (designed to target Pseudomonads) applied to Lolium perenne rhizosphere soil as our test system. The 6-carboxyfluorescein (6-FAM)-PSE1284-hybridised population, defined as displaying enhanced green fluorescence in Flow Cytometry, represented 3.51±1.28% of the total detected population when corrected using a nonsense (NON-EUB338) probe control. Analysis of 16S rRNA gene libraries constructed from Fluorescence Activated Cell Sorted (FACS) -recovered fluorescent populations (n=3), revealed that 98.5% (Pseudomonas spp. comprised 68.7% and Burkholderia spp. 29.8%) of the total sorted population was specifically targeted as evidenced by the homology of the 16S rRNA sequences to the probe sequence. In silico evaluation of probe PSE1284 with the use of RDP-10 probeMatch justified the existence of Burkholderia spp. among the sorted cells. The lack of novelty in Pseudomonas spp. sequences uncovered was notable, probably reflecting the well-studied nature of this functionally important genus. To judge the diversity recorded within the FACS-sorted population, rarefaction and DGGE analysis were used to evaluate, respectively, the proportion of Pseudomonas diversity uncovered by the sequencing effort and the representativeness of the Nycodenz® method for the extraction of bacterial cells from soil.

Tree species diversity interacts with elevated CO2 to induce a greater root system response

As a consequence of land use change and the burning of fossil fuels, atmospheric concentrations of CO2 are increasing and altering the dynamics of the carbon cycle in forest ecosystems. In a number of studies using single tree species, fine root biomass has been shown to be strongly increased by elevated CO2. However, natural forests are often intimate mixtures of a number of co-occurring species. To investigate the interaction between tree mixture and elevated CO2, Alnus glutinosa, Betula pendula and Fagus sylvatica were planted in areas of single species and a three species polyculture in a free-air CO2 enrichment study (BangorFACE). The trees were exposed to ambient or elevated CO2 (580 µmol mol-1) for four years. Fine and coarse root biomass, together with fine root turnover and fine root morphological characteristics were measured. Fine root biomass, and morphology responded differentially to elevated CO2 at different soil depths in the three species when grown in monocultures. In polyculture, a greater response to elevated CO2 was observed in coarse roots to a depth of 20 cm, and fine root area index to a depth of 30 cm. Total fine root biomass was positively affected by elevated CO2 at the end of the experiment, but not by species diversity. Our data suggest that existing biogeochemical cycling models parameterised with data from species grown in monoculture may be underestimating the belowground response to global change.