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In this opinion paper we review recent methodological developments underpinning the study of roots, the rhizosphere and interactions affecting soil functions, and explore new understanding resulting from these advances. We focus on methods that have improved our understanding of rhizodeposition, rhizosphere enzymatic processes and root growth, water and nutrient acquisition at several levels. Finally, we suggest that the future will require new methods that continue to overcome the difficulties posed by the opacity of soil, can scale results spatially and temporally, and integrate multiple aspects of rhizosphere processes simultaneously.

Aims Soil priming affects soil N transformation and plant N availability, but few studies have investigated these interactions to date. Methods To address this, we reviewed the literature for studies quantifying soil priming, soil N transformation and plant N uptake. Results Gross N mineralization was strongly controlled by soil priming in studies with plants, while abiotic factors had a minor influence on gross N mineralization. In contrast, soil priming was negatively related to gross N mineralization and had a low explanatory power in incubation studies where substrates are added as surrogates for root exudates. These results indicate that plants support increased N mineralization and that this is not adequately reflected in incubation studies. Additionally, we observed a positive relationship between soil priming and the % of Norg-derived N uptake as well as total N uptake, which demonstrates that priming enhances the availability of N that was previously organically bound and that at least part of the N mineralized during priming was available for plant uptake. Conclusion Our results show that the effect of roots and rhizodeposition leads to a number of processes supporting N mineralization and availability through priming that are not well reflected in incubation studies. To fully capture the interactions between plant roots and their associated microbiota, we recommend focusing research on systems with plants. Additionally, the strong correlation between C and N transformation should be considered in biogeochemical modelling.

Both plant species and CO₂ concentration can potentially affect rhizodeposition and consequently soil microbial activity and community composition. However, the effect differs based on plant developmental stage. We focused on the effect of three plant species (forbs, grasses, and N₂‐fixers) at an early stage of development on root C deposition and fate, soil organic matter (SOM) mineralization and soil microbial community composition at ambient (aCO₂) and elevated (eCO₂) CO₂ levels. Plants were grown from seed, under continuous ¹³C‐labelling atmospheres (400 and 800 µmol mol⁻¹ CO₂), in grassland soil for three weeks. At the end of the growth period, soil respiration, dissolved organic C (DOC) and phospholipid fatty acid (PLFA) profiles were quantified and isotopically partitioned into root‐ and soil‐derived components. Root‐derived DOC (0.53 ± 0.34 and 0.26 ± 0.29 µg mL soil solution⁻¹) and soil‐derived CO₂ (6.14 ± 0.55 and 5.04 ± 0.44 µg CO₂‐C h⁻¹) were on average two times and 22% higher at eCO₂ than at aCO₂, respectively. Plant species differed in exudate production at aCO₂ (0.11 ± 0.11, 0.10 ± 0.18, and 0.58 ± 0.58 µg mL soil solution⁻¹ for Plantago, Festuca, and Lotus, respectively) but not at eCO₂ (0.20 ± 0.28, 0.66 ± 0.32, and 0.75 ± 0.15 µg mL soil solution⁻¹ for Plantago, Festuca, and Lotus, respectively). However, no differences among plant species or CO₂ levels were apparent when DOC was expressed per gram of roots. Relative abundance of PLFAs did not differ between the two CO₂ levels. A higher abundance of actinobacteria and G‐positive bacteria occurred in unplanted (8.07 ± 0.48 and 24.36 ± 1.18 mol%) and Festuca‐affected (7.63 ± 0.31 and 23.62 ± 0.69 mol%) soil than in Plantago‐ (7.04 ± 0.36 and 23.41 ± 1.13 mol%) and Lotus‐affected (7.24 ± 0.17 and 23.13 ± 0.52 mol%) soil. In conclusion, the differences in root exudate production and soil respiration are mainly caused by differences in root biomass at an early stage of development. However, plant species evidently produce root exudates of varying quality affecting associated microbial community composition.

Purpose Rhizodeposition shapes soil microbial communities that perform important processes such as soil C mineralization, but we have limited understanding of the plant genetic regions influencing soil microbes. Here, barley chromosome regions affecting soil microbial biomass-C (MBC), dissolved organic-C (DOC) and root biomass were characterised. Methods A quantitative trait loci analysis approach was applied to identify barley chromosome regions affecting soil MBC, soil DOC and root biomass. This was done using barley Recombinant Chromosome Substitution Lines (RCSLs) developed with a wild accession (Caesarea 26-24) as a donor parent and an elite cultivar (Harrington) as recipient parent. Results Significant differences in root-derived MBC and DOC and root biomass among these RCSLs were observed. Analysis of variance using single nucleotide polymorphisms genotype classes revealed 16 chromosome regions influencing root-derived MBC and DOC. Of these chromosome regions, five on chromosomes 2H, 3H and 7H were highly significant and two on chromosome 3H influenced both root-derived MBC and DOC. Potential candidate genes influencing root-derived MBC and DOC concentrations in soil were identified. Conclusion The present findings provide new insights into the barley genetic influence on soil microbial communities. Further work to verify these barley chromosome regions and candidate genes could promote marker assisted selection and breeding of barley varieties that are able to more effectively shape soil microbes and soil processes via rhizodeposition, supporting sustainable crop production systems.

Summary • This study examined morphological and topological responses of nodal root axes to defoliation in a fast- and a slow-growing grass species. • Vegetative tillers of both Lolium perenne and Festuca ovina were grown on slant boards and either left intact or subjected to repeated defoliation, under both a high nitrogen (N) and a low N supply. Root length, diameter and branching characteristics were measured on individual nodal root axes. • The total axis root length of F. ovina was less when plants had been defoliated. Root axis weight, primary root axis length and primary root diameter were also less with defoliation than an undefoliated control, under high N. Under low N conditions the root axes of F. ovina had a more randomly branched topology without defoliation. For L. perenne under low N conditions, the length of the primary root axis was longer with defoliation than in an undefoliated control, while the primary root axis diameter decreased. By contrast to F. ovina, the root axes of L. perenne had a more randomly branched topology without defoliation only when supplied with high N. • The greatest plasticity in branching caused by defoliation was observed under high N for L. perenne and under low N for F. ovina. Although grass root axis topology has, in general, a herringbone in structure, the nodal root system can alter root axis structure in response to defoliation.

SummaryAgricultural management practices that increase soil organic matter (SOM), such as no-tillage (NT) with crop residue retention, together with crop varieties best able to source nutrients from SOM may help reverse soil degradation and improve soil nutrient supply and uptake by plants in low-input environments of tropical and sub-tropical areas.Here, we screened germplasm representing genetic diversity within tropical maize breeding programs in relation to shaping SOM mineralisation. Then we assessed effects of contrasting genotypes on nitrification rates, and genotype by management history interactions on these rates.SOM-C mineralisation and gross nitrification rates varied under different maize genotypes. Cumulative SOM-C mineralisation increased with root diameter but decreased with increasing root length. Strong influences of management history and interaction of maize genotype by management history on nitrification were observed. Overall, nitrification rates were higher in NT soil with residue retention.We propose that there is potential to exploit genotypic variation in traits associated with SOM mineralisation and nitrification within breeding programs. Root diameter and length could be used as proxies for root-soil interactions driving these processes. Development of maize varieties with enhanced ability to mineralize SOM combined with NT and residue retention to build/replenish SOM could be key to sustainable production.

Aims The capacity of plant roots to directly acquire organic nitrogen (N) in the form of oligopeptides and amino acids from soil is well established. However, plants have poor access to protein, the central reservoir of soil organic N. Our question is: do plants actively secrete proteases to enhance the breakdown of soil protein or are they functionally reliant on soil microorganisms to undertake this role? Methods Growing maize and wheat under sterile hydroponic conditions with and without inorganic N, we measured protease activity on the root surface (root-bound proteases) or exogenously in the solution (free proteases). We compared root protease activities to the rhizosphere microbial community to estimate the ecological significance of root-derived proteases. Results We found little evidence for the secretion of free proteases, with almost all protease activity associated with the root surface. Root protease activity was not stimulated under N deficiency. Our findings suggest that cereal roots contribute one-fifth of rhizosphere protease activity. Conclusions Our results indicate that plant N uptake is only functionally significant when soil protein is in direct contact with root surfaces. The lack of protease upregulation under N deficiency suggests that root protease activity is unrelated to enhanced soil N capture.

The intrinsic nitrogen (N) supply capacity of soil is central to understanding the productivity of natural plant communities, and essential in the context of determining optimal fertilization rates for agricultural soils. However, it is largely unknown how nutrient availability affects plant mediated priming effects driving soil organic matter mineralisation and associated N-fluxes. We applied continuous, steady-state 13C–CO2 labelling to Lolium perenne grown in high and low productivity grassland soils to allow quantification of SOM- and root-derived soil CO2 efflux. Nutrient treatments (N, P and K) were applied as repeated additions to soils, and impacts on source partitioned soil CO2 efflux were assessed relative to unamended planted and fallow soils. Plants were clipped to uniform height at weekly intervals. Increasing nutrient availability in both soils resulted in a reduction in plant-mediated SOM mineralisation and clipping of plants greatly lowered root-derived respiration but increased SOM mineralisation. Nutrient addition to fallow systems had no effect on SOM mineralisation in either soil. Plant growth stimulated SOM priming, concurrent mobilisation of N from SOM and subsequent plant N uptake in the high productivity soil. Priming was not observed in the low productivity soil due to its greater inherent organic matter stability, resulting in lowered plant-mediated and basal SOM mineralisation. That addition of nutrients reduced SOM mineralisation in planted systems but had no effect in fallow systems is indicative of nutrient availability specifically altering plant-mediated priming of SOM mineralisation. We suggest that plant-soil interactions mediating priming effects are an important determinant of productivity and that the magnitudes of these effects are modified by nutrient availability and soil-specific controls.

Herbivory is one of the most globally distributed disturbances affecting carbon (C)-cycling in trees, yet our understanding of how it alters tree C-allocation to different functions such as storage, growth or rhizodeposition is still limited. Prioritized C-allocation to storage replenishment vs growth could explain the fast recovery of C-storage pools frequently observed in growth-reduced defoliated trees. We performed continuous 13C-labeling coupled to clipping to quantify the effects of simulated browsing on the growth, leaf morphology and relative allocation of stored vs recently assimilated C to the growth (bulk biomass) and non-structural carbohydrate (NSC) stores (soluble sugars and starch) of the different organs of two tree species: diffuse-porous (Betula pubescens Ehrh.) and ring-porous (Quercus petraea [Matt.] Liebl.). Carbon-transfers from plants to bulk and rhizosphere soil were also evaluated. Clipped birch and oak trees shifted their C-allocation patterns above-ground as a means to recover from defoliation. However, such increased allocation to current-year stems and leaves did not entail reductions in the allocation to the rhizosphere, which remained unchanged between clipped and control trees of both species. Betula pubescens and Q. petraea showed differences in their vulnerability and recovery strategies to clipping, the ring-porous species being less affected in terms of growth and architecture by clipping than the diffuse-porous. These contrasting patterns could be partly explained by differences in their C cycling after clipping. Defoliated oaks showed a faster recovery of their canopy biomass, which was supported by increased allocation of new C, but associated with large decreases in their fine root biomass. Following clipping, both species recovered NSC pools to a larger extent than growth, but the allocation of 13C-labeled photo-assimilates into storage compounds was not increased as compared with controls. Despite their different response to clipping, our results indicate no preventative allocation into storage occurred during the first year after clipping in either of the species.

Arbuscular mycorrhizal (AM) hyphae represent an important route for input of plant-derived C to soil, but impacts of these inputs on microbial communities and processes are poorly understood. In this study we characterised pathways of C-flow through microbial communities associated with AM hyphae and quantified impacts on mineralisation of native SOM. Continuous, steady-state 13CO2 labelling was applied throughout the growth period (60 d) of Lolium perenne. Exclusion meshes were used to control access of roots and AM hyphae to soil, and plant-derived C was quantified within microbial PLFA and NLFA, and soil CO2 efflux was partitioned into plant- and soil organic matter (SOM) derived components. Pathways of C-flow through hyphosphere and mycorrhizosphere communities were distinct, as was the fate of plant-derived C from AM hyphae accessing soil through 37 and 1 μm meshes. Mineralisation of native SOM was increased in all treatments, relative to unplanted controls, and this priming effect was largest for AM hyphae accessing soil through the 1 μm mesh size. We demonstrated that AM hyphae can strongly increase mineralisation of native SOM and identified distinct pathways of C-flow through hyphosphere communities. Our results suggest that, in addition to affecting rates of litter decomposition, AM hyphae may have a significant influence on turnover of native SOM.

Phospholipid fatty acid (PLFA) analysis is an effective non-culture-based technique for providing information on the living soil microbial community. The coupling of 13C tracers with PLFA analysis can indicate the response of microbial populations to environmental change and has been widely used to trace C flux in soil-plant systems. Based on studies applying 13C PLFA analysis, the current technological status, current applications and future opportunities are discussed and evaluated. First we describe some aspects of the labelling and analytical methodology. The approaches to study the incorporation of 13C substrate and rhizodeposition C into soil microbial communities are compared. We continue with the application of 13C-labelling to study soil microbial communities, including the utilization of soil mineralisation products, the C flux from plants into the soil microbial pool, the biodegradation of pollutants and on the application to a specific microbial group, i.e. methanotrophs. Additionally, some perspectives on the limitations of the 13C PLFA method and future research avenues are noted. Although including some limitations and complications, the 13C PLFA method provides an excellent tool for understanding the relationship between microbial populations and soil biogeochemical cycling, thus providing a key to open the soil microbial black box.

34 páginas, 2 tablas, 4 figuras, [EN] Current knowledge on tree carbon (C) allocation to wood is particularly scarce in plants subjected to disturbance factors, such as browsing, which affects forest regeneration worldwide and has an impact on the C balance of trees. Furthermore, quantifying the degree to which tree rings are formed from freshly assimilated vs. stored carbohydrates is highly relevant for our understanding of tree C allocation. We used (13)C labelling to quantify seasonal allocation of stored C to wood formation in two species with contrasting wood anatomy: Betula pubescens Ehrh. (diffuse-porous) and Quercus petraea [Matt.] Liebl. (ring-porous). Clipping treatments (66% shoot removal, and unclipped) were applied to analyse the effect of browsing on C allocation into tree rings, plus the effects on tree growth, architecture, ring width and non-structural carbohydrates (NSCs). The relative contribution of stored C to wood formation was greater in the ring-porous (55-70%) than in the diffuse-porous species (35-60%), although each species followed different seasonal trends. Clipping did not cause a significant depletion of C stores in either species. Nonetheless, a significant increase in the proportion of stored C allocated to earlywood growth was observed in clipped birches, and this could be explained through changes in tree architecture after clipping. The size of C pools across tree species seems to be important in determining the variability of seasonal C allocation patterns to wood and their sensibility to disturbances such as browsing. Our results indicate that the observed changes in C allocation to earlywood in birch were not related to variations in the amount or concentration of NSC stores, but to changes in the seasonal availability of recently assimilated C caused by modifications in tree architecture after browsing., S.P. was funded by an MEC-SEUI postdoctoral fellowship and a JAE-Doc (CSIC) contract. This study was partly funded by projects CGL2008-04847-C02-01/BOS (MCI-Spain) and GA-LC- 011/2008 (DGA-La Caixa). E.P., A.S., A.H. and P.M. thank the Scottish Government’s Rural and Environmental Research and Analysis Directorate for funding.

For soils in carbon balance, losses of soil carbon from biological activity are balanced by organic inputs from vegetation. Perturbations, such as climate or land use change, have the potential to disrupt this balance and alter soil-atmosphere carbon exchanges. As the quantification of soil organic matter stocks is an insensitive means of detecting changes, certainly over short timescales, there is a need to apply methods that facilitate a quantitative understanding of the biological processes underlying soil carbon balance. We outline the processes by which plant carbon enters the soil and critically evaluate isotopic methods to quantify them. Then, we consider the balancing CO(2) flux from soil and detail the importance of partitioning the sources of this flux into those from recent plant assimilate and those from native soil organic matter. Finally, we consider the interactions between the inputs of carbon to soil and the losses from soil mediated by biological activity. We emphasize the key functional role of the microbiota in the concurrent processing of carbon from recent plant inputs and native soil organic matter. We conclude that quantitative isotope labelling and partitioning methods, coupled to those for the quantification of microbial community substrate use, offer the potential to resolve the functioning of the microbial control point of soil carbon balance in unprecedented detail.

A combined proteomic and isotope tracer approach was used to investigate the impact of supplying N as glycine to roots of Lolium perenne. Initially, ammonium nitrate was supplied to all plants, after which half received glycine as their sole N source, while the remainder continued to receive ammonium nitrate. Plants supplied with glycine acquired less N than those receiving the mineral source, resulting in reduced root nitrate concentrations. The amino acid complement of roots was also strongly affected by the form of N supplied, and 15 N labelling indicated that the biochemical fate of acquired N in roots was dependent on the form of N available for uptake. Proteomic analysis of Lolium roots indicated that 6% of 627 root proteins resolved on 2D gels changed in abundance in response to the form of N applied. Multivariate analysis of protein abundance clearly discriminated the proteomes of L. perenne roots as a function of treatment applied. Seven affected proteins were identified (mostly by protein homology with sequenced species), including methionine adenosyltransferase, an enzyme involved in glycine metabolism. Although some changes in root amino acid and protein complements were due to responses to reduced N supply, both the distinct fate of 15 N tracers and the abundances of identified proteins could be attributed specifically to the form of N available to roots. The results demonstrate the potential of targeted proteomic approaches to identify functioning of plants where more traditional methods cannot resolve multiple, co-incident biological interactions and element fluxes.

Intercropping is a farming practice involving two or more crop species, or genotypes, growing together and coexisting for a time. On the fringes of modern intensive agriculture, intercropping is important in many subsistence or low-input/resource-limited agricultural systems. By allowing genuine yield gains without increased inputs, or greater stability of yield with decreased inputs, intercropping could be one route to delivering ‘sustainable intensification’. We discuss how recent knowledge from agronomy, plant physiology and ecology can be combined with the aim of improving intercropping systems. Recent advances in agronomy and plant physiology include better understanding of the mechanisms of interactions between crop genotypes and species – for example, enhanced resource availability through niche complementarity. Ecological advances include better understanding of the context-dependency of interactions, the mechanisms behind disease and pest avoidance, the links between above- and below-ground systems, and the role of microtopographic variation in coexistence. This improved understanding can guide approaches for improving intercropping systems, including breeding crops for intercropping. Although such advances can help to improve intercropping systems, we suggest that other topics also need addressing. These include better assessment of the wider benefits of intercropping in terms of multiple ecosystem services, collaboration with agricultural engineering, and more effective interdisciplinary research.

International audience; The objective of this work was to determine if the impact of nitrogen (N) on the release of organic carbon (C) into the soil by roots (rhizodeposition) correlated with the effect of this nutrient on some variables of plant growth. Lolium multiflorum Lam. was grown at two levels of N supply, either in sterile sand percolated with nutrient solution or in non-sterile soil. The axenic sand systems allowed continuous quantification of rhizodeposition and accurate analysis of root morphology whilst the soil microcosms allowed the study of C-14 labelled C flows in physico-chemical and biological conditions relevant to natural soils. In the axenic sand cultures, enhanced N supply strongly increased the plant biomass, the plant N content and the shoot to root ratio. N supply altered the root morphology by increasing the root surface area and the density of apices, both being significantly positively correlated with the rate of organic C release by plant roots before sampling. This observation is consistent with the production of mucilage by root tips and with mechanisms of root exudation reported previously in the literature, i.e. the passive diffusion of roots solutes along the root with increased rate behind the root apex. We proposed a model of root net exudation, based on the number of root apices and on root soluble C that explained 60% of the variability in the rate of C release from roots at harvest. The effects of N on plant growth were less marked in soil, probably related to the relatively high supply of N from non-fertiliser soil-sources. N fertilization increased the shoot N concentration of the plants and the shoot to root ratio. Increased N supply decreased the partitioning of C-14 to roots. In parallel, N fertilisation increased the root soluble C-14 and the C-14 recovered in the soil per unit of root biomass, suggesting a stimulation of root exudation by N supply. However, due to the high concentration of N in our unfertilised plants, this stimulation was assumed to be very weak because no significant effect of N was observed on the microbial C and on the bacterial abundance in the rhizosphere. Considering the difficulties in evaluating rhizodeposition in non sterile soil, it is suggested that the root soluble C, the root surface area and the root apex density are additional relevant variables that should be useful to measure along with the variables that are commonly determined when investigating how plant functioning impacts on the release of C by roots (i.e soil C, C of the microbial biomass, rhizosphere respiration).

Coupling growth of Lolium perenne L. in sterile solution culture with steady-state 13CO2 labelling allowed quantification of the contribution of C, assimilated either before or after a specific time point, both to plant compartments and root exudates. Plants were grown for 27 days in atmospheres containing CO2 with δ13C signatures of either −13.5 or −36.1‰. Air supplies to plants were then reciprocally switched to the opposing signature (day 0), plants were destructively harvested and root exudates collected over the next 8 days. Following the switch, C assimilated after day 0 and transported to the roots initially only appeared in root tips, later appearing in both tip and non-tip material. The δ13C signature of the root exudate changed exponentially with time. Assimilation pre- and post-day 0 contributed equally to exudate C at 4.5 days. Beyond day 8, assimilation pre-day 0 still contributed 41.7% of exudate C. Over all 8 days, a linear relationship existed between the δ13C signatures of root tips and exudate, suggesting that all newly assimilated C in the exudate was from root tips. Results imply pulse-labelling approaches to study root exudates are discriminative in the sources of exudates labelled and in the sites from which exudation occurs.

The aim of this study was to investigate the physiological basis of increased root exudation from Festuca rubra, in response to defoliation. The hypothesis, that assimilate supply to roots is a key determinant of the response of root exudation to defoliation was tested by imposing CO2-deplete (< 50 μmol mol−1) atmospheres to F. rubra. This was done as a non-destructive means of preventing supply of new assimilate to roots of intact and defoliated plants. F. rubra was grown in axenic sand systems, with defoliation and CO2-depletion treatments applied to plants at 14 and 35 days after planting. Root exudation and NO3 − uptake were quantified throughout, and post-treatment uptake and allocation of N were determined from the distribution of 15N label, supplied as 15NO3 −. Defoliation of F. rubra resulted in significantly (P

In defoliated grasses, where photosynthesis is reduced due to removal of leaf material, it is well established that remobilization of nitrogen occurs from both older remaining leaves and roots towards the younger growing leaves. In contrast, little is known about the movement of nitrogen within intact grass plants experiencing prolonged inhibition of photosynthesis. We tested the following hypotheses in Festuca rubra L. ssp. rubra cv. Boreal: that both reduction of the atmospheric CO 2 concentration and defoliation (1) induce mobilization of nitrogen from roots and older leaves towards growing leaves and (2) elicit similar directional change in the abundance of proteins in roots and older leaves relevant to the process of nitrogen mobilization including, glutamine synthetase (GS), EC 6.3.1.2; papain, EC 3.4.22.2; chymopapain, EC 3.4.22.6; ribulose bisphosphate carboxylaseloxygenase (Rubisco), EC 4.1.1.39; and the light harvesting complex of photosystem II (LHCPII). After growth at ambient atmospheric CO 2 concentration, plants of E rubra were subject to atmospheres containing either ambient (350 μl l -1 ) or deplete (

The aim of this study was to determine the effects of N-supply and defoliation on rhizodeposition from Festuca rubra, in the context of whole-plant C- partitioning and root morphology. Plants were grown for 36 d in axenic sand microcosms continuously percolated with nutrient solutions of either high or low N concentration (2 mM or 0.01 mM NH(4)NO(3), respectively). The effects of partial defoliation at weekly intervals were determined at high and low N. At low N, dry matter accumulation in roots and shoots was reduced significantly (P

This study investigated the effects of N-supply and partial defoliation on C-partitioning, root morphology and soluble rhizodeposition, for Lolium perenne grown in axenic sand culture systems percolated with nutrient solution. Plants were grown for 36 d in nutrient solutions with differing N concentrations (4 mM or 0.02 mM NH4 +NO3 -), and effects of repeated defoliation to 4 cm were determined. The ‘low N’ supply reduced (P < 0.05) dry matter accumulation, with proportionately increased partitioning to the root systems. Root morphology was also altered at ‘low N’, with development of a finer root system, manifest as increased (P < 0.05) specific root length. Concurrent with these effects on growth of L. perenne, ‘low N’ increased (P < 0.05) exudation of C-compounds from roots on a per g root basis. Defoliation was found to increase exudation (P < 0.05) of soluble compounds for periods of 3-5 d following each cut, at both N-supply rates. The effects of N-supply and defoliation are of importance in understanding the coupling of plant productivity to nutrient cycling in soils with differing N availabilities and for grassland systems which are subject to grazing.

The distribution and partitioning of dry matter and photoassimilate of Lolium perenne was investigated under two light regimes providing photosynthetically active radiation of 350 /imo\ m 2 s~1 (low light treatment) or 1000 //mol m 2 s~1 (high light treatment). Plants were grown at specific growth conditions in either soil or sand microcosm units to follow the subsequent release of carbon into the rhizosphere and its consequent incorporation into the microbial biomass (soil system) or recovery as exudates (sand system). The distribution of recent assimilate between the plant and root released carbon pools was determined using 14C02 pulse-chase methodology at both light treatments and for both sand- and soil-grown seedlings. A significant (P

Plant and Soil, 368 (1-2), ISSN:0032-079X, ISSN:1573-5036

Summary • The overstorey coniferous trees and understorey ericaceous dwarf shrubs of northern temperate and boreal forests have previously been considered to form mycorrhizas with taxonomically and functionally distinct groups of fungi.  Here, we tested the hypothesis that Meliniomyces variabilis and Meliniomyces bicolor, isolated from Piceirhiza bicolorata ectomycorrhizas of pine, can function as ericoid mycorrhizal symbionts with Vaccinium vitis-idaea. We used split-compartment microcosms to measure the reciprocal exchange of 13 C and 15 N between V. vitisidaea and three fungal isolates in the Hymenoscyphus ericae aggregate isolated from Scots pine ectomycorrhizas (M. variabilis and M. bicolor) or Vaccinium roots (M. variabilis).  The extramatrical fungal mycelium of labelled mycorrhizal plants was significantly enriched in 13 C, and the leaves were significantly enriched in 15 N, compared with nonmycorrhizal and nonlabelled controls.  These findings show for the first time that fungi in the H. ericae aggregate, isolated from pine ectomycorrhizas, can transfer C and N and can thus form functional ericoid mycorrhizas in an understorey ericaceous shrub.

The aims of the study were to determine group specificity in microbial utilization of root-exudate compounds and whole rhizodeposition; quantify the proportions of carbon acquired by microbial groups from soil organic matter and rhizodeposition, respectively; and assess the importance of root-derived C as a driver of soil microbial community structure. Additions of 13C-labelled root-exudate compounds to organic soil and steady-state labelling of Lolium perenne, coupled to compound-specific isotope ratio mass spectrometry, were used to quantify group-specific microbial utilization of rhizodeposition. Microbial utilization of glucose and fumaric acid was widespread through the microbial community, but glycine was utilized by a narrower range of populations, as indicated by the enrichment of phospholipid fatty acid (PLFA) analysis fractions. In L. perenne rhizospheres, high rates of rhizodeposit utilization by microbial groups showed good correspondence with increased abundance of these groups in the rhizosphere. Although rhizodeposition was not the quantitatively dominant C source for microbes in L. perenne rhizospheres, relative utilization of this C source was an important driver of microbial group abundance in organic soil.

Root proliferation as a response to exploit zones of nutrient enrichment in soil has been demonstrated for a wide range of plant species. However, the effectiveness of this as a strategy to acquire nutrients is also dependent on interactions with the soil microbial community. Specifically, C-flow from roots modifies microbial activity and probably the balance between nutrient mineralization and immobilization processes in the rhizosphere. In this study, near-natural abundance 13C-labelling and gene-reporter methods were applied to determine the effects of uneven nitrate supply to roots of Hordeum vulgare on assimilate partitioning and root exudation. Plants were initially grown in uniform nitrate supply in split-root, sand microcosms after which one treatment continued to receive uniform supply, and the other received nitrate to one root compartment only. At the time of imposing the treatments, the CO2 supplied to the plants was switched to a cylinder source, providing a distinct delta13C-signature and allowing the fate of new assimilate within the plants to be determined. The labelling approach allowed quantification of the expected preferential allocation of new C-assimilate to roots in enriched nitrate, prior to any measurable effect on whole biomass or root architecture. Biosensor (lux-marked Pseudomonas fluorescens 10586 pUCD607) bioluminescence, quantified spatially by CCD imaging, demonstrated that root exudation was significantly increased for roots in enriched nitrate. This response of root exudation, being primarily associated with root apices and concurrent with enhanced assimilate supply, strongly suggests that C-flow from roots is an integral component of the proliferation response to nitrate.

The aim of this study was to determine the relationship between shoot nitrate concentration, mediated by nitrate supply to roots, and root exudation from Hordeum vulgare. Plants were grown for 14 d in Cfree sand microcosms, supplied with nutrient solution containing 2 mM nitrate. After this period, three treatments were applied for a further 14 d: (A) continued supply with 2 mM nitrate (zero boost), (B) supply with 10 mM nitrate (low boost), and (C) supply with 20 mM nitrate (high boost). At the end of the treatment period, a bacterial biosensor (Pseudomonas fluorescens 10586 pUCD607, marked with the lux CDABE genes for bioluminescence) was applied to the microcosms to report on C-substrate availability, as a consequence of root exudation. The nitrate boost treatments significantly affected shoot nitrate concentrations, in the order C>B>A. In treatments receiving a nitrate boost (B, C), increased shoot nitrate concentration was correlated with increased plant biomass, reduced root length, reduced number of root tips, and increased mean root diameter, relative to the no boost treatment (A). Imaging of biosensor bioluminescence (proportional to metabolic activity in response to availability of root exudates) indicated that root exudation increased with decreasing shoot nitrate concentration. Biosensor reporting of root C-flow indicated that exudation was greater from root tip regions than from the whole root, but that specific exudation rates for all sites were unaffected by treatments. Total root exudation across treatments was found to be closely correlated with total root length, indicating that increased root exudation, per unit root biomass, with decreasing nitrate supply was associated with altered root morphology, as a consequence of systemic plant responses to internal N-status.

The deposition of organic compounds from plant roots is a key determinant of rhizosphere microbial activity and community structure. Consequently, C-flow from roots to soil is an important process in coupling plant and microbial productivity, via impacts on microbial nutrient cycling in soil. Experimentally, isotopic tracers ( 13 C or 14 C) are used to track C inputs to soil and microbial communities. However, in many such studies the relationship between labelled Cflows and total C-flows are not established, limiting the interpretative value of the results. In this study, we applied steady-state near natural abundance 13 CO 2 labelling to determine the impact of partial defoliation of Festuca rubra on root exudation. This approach in axenic culture facilitated determination of the contribution of pre- and post-defoliation assimilates both to root C-flow and plant tissues. The results demonstrated that total root exudation was increased in the two days following defoliation. This was concurrent with reduced net CO 2 assimilation and reduced allocation of post-defoliation assimilates belowground and to active root meristems. Through determination of the d d d 13 C of root exudates, it was established that the source of the increased root exudation was pre-defoliation assimilate. However, this response was transient, with reduced deposition of pre- and post-defoliation assimilates from roots during the period 2‐4 d following defoliation. The results highlight the limitations of pulse-labelling approaches as a means of quantifying impacts of treatments on root exudation, particularly where the treatment is likely to affect plant C-partitioning or the balance between deposition to, and re-mobilization from, C-storage pools.

Background and aims Plant-bacterial interactions in the rhizosphere are important in mediating soil nutrient transformations. Plants supply carbon-rich substrates to rhizobacteria as root exudates and bacteria mobilise soil-bound phosphate for plant nutrition. This study aimed to probe the specificity of the plant effect on bacterial gene expression in Pstarved rhizosphere conditions. Methods DNA microarrays were employed to study gene expression in the rhizosphere of Lolium perenne grown under high and low phosphate regimes (330 ?M vs. 3-6 ?M phosphate). Root exudation under these regimes was also quantified. Phosphateregulated gene expression of a panel of 22 genes was compared in rhizosphere, planktonic culture and during biofilm growth on an artificial root. Results Plant growth and root exudation were affected by P-availability. P-limited conditions induced increased expression of bacterial genes of an aromatic degradation pathway (catA), heavy metal sensing (PA2523), and membrane proteins (glpM, crcB), while genes involved in cell motility and amino acid uptake/ metabolism were downregulated. A crcB mutant was impaired in rhizosphere survival under low phosphate conditions, though glpM and catA mutants were not affected. Several of the genes studied were induced by phosphate limitation in all three lifestyles studied. Conclusions Our results show the importance of the plant-microbe interaction in controlling the bacterial transcriptional response in a phosphate-limited rhizosphere.