Susan J. Grayston

Rhizosphere carbon flow in trees, in comparison with annual plants: the importance of root exudation and its impact on microbial activity and nutrient availability

Abstract The release of organic substances from roots is a key process influencing nutrient availability in the rhizosphere. Rhizodeposition, including root exudation can influence plant growth directly by making cations available for uptake through processes such as chelation or indirectly by influencing soil microbial activity. It is important to gain knowledge about the range of compounds released and the factors influencing their release, to understand their effects on the microbial community and enable development of techniques to enhance microbial activity. The increasing growth of trees in various land use systems is coupled with a limited knowledge of the interactions between nutrient availability and tree growth. This highlights the need for a greater understanding of factors affecting nutrient availability in these systems. The purpose of this paper is to review the various strategies which are used to measure rhizodeposition by plants and demonstrate that root exudates are an important component of carbon loss from plants and that they may have a more important role in nutrient acquisition and plant growth than previously thought. The paper will discuss the character of carbon loss from trees in comparison to annual plants and discuss the increasing evidence of the importance of non-nutrient components of root exudates as host specific recognition signals. The factors affecting exudate release and the impact of these compounds on nutrient availability will be discussed. The limitations of previous studies of rhizodeposition and root exudation through omission of a mycorrhizal component, and the need for further research in this neglected area, will be highlighted. Manipulation of plant-microbial interactions is discussed in relation to improving or maintaining plant growth in sustainable systems.

Assessing shifts in microbial community structure across a range of grasslands of differing management intensity using CLPP, PLFA and community DNA techniques.

This study aimed to characterise soil microbial community structure and function in temperate upland grassland ecosystems. We compared the use of community level physiological profiles (CLPP), phospholipid fatty acid (PLFA) profiles and community DNA (%G+C base distribution) approaches to quantify soil microbial community structure and potential activity across a gradient of three upland grassland types at 10 geographically distinct sites within the UK. Soil microbial biomass (Cmic) was highest in unimproved (U4a) and lowest in improved (MG6) grasslands. In contrast, potential soil microbial activity (carbon utilisation) was greatest in the improved and lowest in the semi-improved (U4b) grasslands. PLFA and culturing revealed that the soil microbial community shifted from one favouring fungi to one favouring bacteria as grassland improvement increased. Canonical variate analysis (CVA) of the CLPP and PLFA data differentiated microbial communities from the grassland types and sites and the separation between grasslands was greater using PLFA than CLPP. Discrimination between grasslands was mainly due to the presence of higher concentrations of fatty acids typical for Gram −ve bacteria in improved grasslands and actinomycete and fungal fatty acids in the semi and unimproved grasslands. CVA of the %G+C data gave less discrimination of the microbial communities than the other two methods. Correlation analysis of the CVA data for each microbial analysis showed a small, but significant, level of matching between the CLPP and PLFA data suggesting these two analyses may be reporting on similar members of the microbial community. Correlation between microbial community structure and soil physio-chemical properties indicated that PLFA were highly correlated with calcium, phosphorus, sodium, nitrogen and organic matter content and pH. CLPP were highly correlated with sodium and organic matter content and pH, while %G+C content correlated with pH. Correlation between microbial community structure and plant community structure indicated that fatty acids typical for Gram −ve bacteria were highly correlated with the presence of Lolium perenne and Trifolium repens and all microbial PLFA with the presence of Vaccinium myrtillus. Correlation of plant species with CLPP indicated that the presence of a number of rushes, shrubs, herbs and grasses influenced the metabolic profiles of the microbial communities from these grasslands. The presence of herbs were found to be highly correlated with certain %G+C classes within the community DNA.

Microbial community structure and characteristics of the organic matter in soils under Pinus sylvestris, Picea abies and Betula pendula at two forest sites

Abstract Microbial biomass C (Cmic), C mineralization rate, phospholipid fatty acid (PLFA) profiles and community level physiological profiles (CLPPs) using Biolog were determined from the humus and mineral soil layers in adjacent stands of Scots pine (Pinus sylvestris L.), Norway spruce [Picea abies (L.) Karst.] and silver birch (Betula pendula Roth) at two forest sites of different fertility. In addition, the Fourier-transformed infrared (FTIR) spectra were run on the samples for characterization of the organic matter. Cmic and C mineralization rate tended to be lowest under spruce and highest under birch, at the fertile site in all soil layers and at the less fertile site in the humus layer. There were also differences in microbial community structure in soils under different tree species. In the humus layer the PLFAs separated all tree species and in the mineral soil spruce was distinct from pine and birch. CLPPs did not distinguish microbial communities from the different tree species. The FTIR spectra did not separate the tree species, but clearly separated the two sites.

EFFECTS OF GRAZING ON MICROBIAL FUNCTIONAL GROUPS INVOLVED IN SOIL N DYNAMICS

Enhancement of soil nitrogen (N) cycling by grazing has been observed in many grassland ecosystems. However, whether grazing affects the activity only of the key microbial functional groups driving soil N dynamics or also affects the size (cell number) and/or composition of these groups remains largely unknown. We studied the enzyme activity, size, and composition of five soil microbial communities (total microbial and total bacterial communities, and three functional groups driving N dynamics: nitrifiers, denitrifiers, and free N2 fixers) in grassland sites experiencing contrasting sheep grazing regimes (one light grazing [LG] site and one intensive grazing [IG] site) at two topographical locations. Enzyme activity was determined by potential carbon mineralization, nitrification, denitrification, and N2 fixation assays. The size of each community (except N2 fixers) was measured by the most-probable-number technique. The composition of the total soil microbial community was characterized by phospholipid f...

Spatial structure in soil chemical and microbiological properties in an upland grassland

We characterised the spatial structure of soil microbial communities in an unimproved grazed upland grassland in the Scottish Borders. A range of soil chemical parameters, cultivable microbes, protozoa, nematodes, phospholipid fatty acid (PLFA) profiles, community-level physiological profiles (CLPP), intra-radical arbuscular mycorrhizal community structure, and eubacterial, actinomycete, pseudomonad and ammonia-oxidiser 16S rRNA gene profiles, assessed by denaturing gradient gel electrophoresis (DGGE) were quantified. The botanical composition of the vegetation associated with each soil sample was also determined. Geostatistical analysis of the data revealed a gamut of spatial dependency with diverse semivariograms being apparent, ranging from pure nugget, linear and non-linear forms. Spatial autocorrelation generally accounted for 40-60% of the total variance of those properties where such autocorrelation was apparent, but accounted for 97% in the case of nitrate-N. Geostatistical ranges extending from approximately 0.6-6 m were detected, dispersed throughout both chemical and biological properties. CLPP data tended to be associated with ranges greater than 4.5 m. There was no relationship between physical distance in the field and genetic similarity based on DGGE profiles. However, analysis of samples taken as close as 1 cm apart within a subset of cores suggested some spatial dependency in community DNA-DGGE parameters below an 8 cm scale. Spatial correlation between the properties was generally weak, with some exceptions such as between microbial biomass C and total N and C. There was evidence for scale-dependence in the relationships between properties. PLFA and CLPP profiling showed some association with vegetation composition, but DGGE profiling did not. There was considerably stronger association between notional sheep urine patches, denoted by soil nutrient status, and many of the properties. These data demonstrate extreme spatial variation in community-level microbiological properties in upland grasslands, and that despite considerable numeric ranges in the majority of properties, overarching controlling factors were not apparent.

Characterization of Humus Microbial Communities in Adjacent Forest Types That Differ in Nitrogen Availability

To address the link between soil microbial community composition and soil processes, we investigated the microbial communities in forest floors of two forest types that differ substantially in nitrogen availability. Cedar-hemlock (CH) and hemlock-amabilis fir (HA) forests are both common on northern Vancouver Island, B.C., occurring adjacently across the landscape. CH forest floors have low nitrogen availability and HA high nitrogen availability. Total microbial biomass was assessed using chloroform fumigation-extraction and community composition was assessed using several cultivation-independent approaches: denaturing gradient gel electrophoresis (DGGE) of the bacterial communities, ribosomal intergenic spacer analysis (RISA) of the bacterial and fungal communities, and phospholipid fatty acid (PLFA) profiles of the whole microbial community. We did not detect differences in the bacterial communities of each forest type using DGGE and RISA, but differences in the fungal communities were detected using RISA. PLFA analysis detected subtle differences in overall composition of the microbial community between the forest types, as well as in particular groups of organisms. Fungal PLFAs were more abundant in the nitrogen-poor CH forests. Bacteria were proportionally more abundant in HA forests than CH in the lower humus layer, and Gram-positive bacteria were proportionally more abundant in HA forests irrespective of layer. Bacterial and fungal communities were distinct in the F, upper humus, and lower humus layers of the forest floor and total biomass decreased in deeper layers. These results indicate that there are distinct patterns in forest floor microbial community composition at the landscape scale, which may be important for understanding nutrient availability to forest vegetation.

Recreating a Functioning Forest Soil in Reclaimed Oil Sands in Northern Alberta: An Approach for Measuring Success in Ecological Restoration

During oil-sands mining all vegetation, soil, overburden, and oil sand is removed, leaving pits several kilometers wide and up to 100 m deep. These pits are reclaimed through a variety of treatments using subsoil or a mixed peat-mineral soil cap. Using nonmetric multidimensional scaling and cluster analysis of measurements of ecosystem function, reclamation treatments of several age classes were compared with a range of natural forest ecotypes to discover which treatments had created ecosystems similar to natural forest ecotypes and at what age this occurred. Ecosystem function was estimated from bioavailable nutrients, plant community composition, litter decomposition rate, and development of a surface organic layer. On the reclamation treatments, availability of nitrate, calcium, magnesium, and sulfur were generally higher than in the natural forest ecotypes, while ammonium, P, K, and Mn were generally lower. Reclamation treatments tended to have more bare ground, grasses, and forbs but less moss, lichen, shrubs, trees, or woody debris than natural forests. Rates of litter decomposition were lower on all reclamation treatments. Development of an organic layer appeared to be facilitated by the presence of shrubs. With repeated applications of fertilizers, measured variables for the peat-mineral amendments fell within the range of natural variability at about 20 yr. An intermediate subsoil layer reduced the need for fertilizer and conditions resembling natural forests were reached about 15 yr after a single fertilizer application. Treatments over tailings sand receiving only one application of fertilizer appeared to be on a different trajectory to a novel ecosystem.

Evidence of a strong coupling between root exudation, C and N availability, and stimulated SOM decomposition caused by rhizosphere priming effects

Increased temperatures and concomitant changes in vegetation patterns are expected to dramatically alter the functioning of northern ecosystems over the next few decades. Predicting the ecosystem response to such a shift in climate and vegetation is complicated by the lack of knowledge about the links between aboveground biota and belowground process rates. Current models suggest that increasing temperatures and rising concentrations of atmospheric CO2 will be partly mitigated by elevated C sequestration in plant biomass and soil. However, empirical evidence does not always support this assumption, as elevated temperature and CO2 concentrations also accelerate the belowground C flux, in many cases extending to increased decomposition of soil organic matter (SOM) and ultimately resulting in decreased soil C stocks. The mechanism behind the increase has remained largely unknown, but it has been suggested that priming might be the causative agent. Here, we provide quantitative evidence of a strong coupling between root exudation, SOM decomposition, and release of plant available N caused by rhizosphere priming effects. As plants tend to increase belowground C allocation with increased temperatures and CO2 concentrations, priming effects need to be considered in our long-term analysis of soil C budgets in a changing environment. The extent of priming seems to be intimately linked to resource availability, as shifts in the stoichiometric nutrient demands of plants and microorganisms will lead to either cooperation (resulting in priming) or competition (no priming will occur). The findings lead us on the way to resolve the varying response of primary production, SOM decomposition, and release of plant available N to elevated temperatures, CO2 concentrations, and N availability.

Discontinuity in the responses of ecosystem processes and multifunctionality to altered soil community composition

Significance Ecosystem functioning is more strongly affected by biodiversity loss when multiple functions are considered because different species affect different functions. To quantify these biodiversity-functioning relationships, the emerging multifunctionality framework advocates calculation of indices that aggregate responses of individual functions. Data aggregation, however, is notorious for providing misleading information by obscuring true relationships between explanatory and response variables. We test the ability of common multifunctionality indices to reveal effects on key ecosystem functions of changes in soil communities. The multifunctionality indices all decrease with soil animal loss, but the responses of individual functions diverge markedly from these aggregated metrics. Application of the multifunctionality framework for landscape provision of multiple ecosystem services should therefore emphasize understanding relationships between communities and individual functions. Ecosystem management policies increasingly emphasize provision of multiple, as opposed to single, ecosystem services. Management for such “multifunctionality” has stimulated research into the role that biodiversity plays in providing desired rates of multiple ecosystem processes. Positive effects of biodiversity on indices of multifunctionality are consistently found, primarily because species that are redundant for one ecosystem process under a given set of environmental conditions play a distinct role under different conditions or in the provision of another ecosystem process. Here we show that the positive effects of diversity (specifically community composition) on multifunctionality indices can also arise from a statistical fallacy analogous to Simpson’s paradox (where aggregating data obscures causal relationships). We manipulated soil faunal community composition in combination with nitrogen fertilization of model grassland ecosystems and repeatedly measured five ecosystem processes related to plant productivity, carbon storage, and nutrient turnover. We calculated three common multifunctionality indices based on these processes and found that the functional complexity of the soil communities had a consistent positive effect on the indices. However, only two of the five ecosystem processes also responded positively to increasing complexity, whereas the other three responded neutrally or negatively. Furthermore, none of the individual processes responded to both the complexity and the nitrogen manipulations in a manner consistent with the indices. Our data show that multifunctionality indices can obscure relationships that exist between communities and key ecosystem processes, leading us to question their use in advancing theoretical understanding—and in management decisions—about how biodiversity is related to the provision of multiple ecosystem services.

Effects of long-term fertilization of forest soils on potential nitrification and on the abundance and community structure of ammonia oxidizers and nitrite oxidizers

Forest fertilization in British Columbia is increasing, to alleviate timber shortfalls resulting from the mountain pine beetle epidemic. However, fertilization effects on soil microbial communities, and consequently ecosystem processes, are poorly understood. Fertilization has contrasting effects on ammonia-oxidizing bacteria and archaea (AOB and AOA) in grassland and agricultural ecosystems, but there are no studies on AOB and AOA in forests. We assessed the effect of periodic (6-yearly application 200 kg N ha⁻¹) and annual (c. 75 kg N ha⁻¹) fertilization of lodgepole pine and spruce stands at five long-term maximum productivity sites on potential nitrification (PN), and the abundance and diversity of AOB, AOA and Nitrobacter and Nitrospira-like nitrite-oxidizing bacteria (NOB). Fertilization increased AOB and Nitrobacter-like NOB abundances at some sites, but did not influence AOA and Nitrospira-like NOB abundances. AOB and Nitrobacter-like NOB abundances were correlated with PN and soil nitrate concentration; no such correlations were observed for AOA and Nitrospira-like NOB. Autotrophic nitrification dominated (55–97%) in these forests and PN rates were enhanced for up to 2 years following periodic fertilization. More changes in community composition between control and fertilized plots were observed for AOB and Nitrobacter-like NOB than AOA. We conclude that fertilization causes rapid shifts in the structure of AOB and Nitrobacter-like NOB communities that dominate nitrification in these forests.