Carbon availability affects already large species-specific differences in chemical composition of ectomycorrhizal fungal mycelia in pure culture.

Although ectomycorrhizal (ECM) contribution to soil organic matter processes receives increased attention, little is known about fundamental differences in chemical composition among species, and how that may be affected by carbon (C) availability. Here, we study how 16 species (incl. 19 isolates) grown in pure culture at three different C:N ratios (10:1, 20:1, and 40:1) vary in chemical structure, using Fourier transform infrared (FTIR) spectroscopy. We hypothesized that C availability impacts directly on chemical composition, expecting increased C availability to lead to more carbohydrates and less proteins in the mycelia. There were strong and significant effects of ECM species (R2 = 0.873 and P = 0.001) and large species-specific differences in chemical composition. Chemical composition also changed significantly with C availability, and increased C led to more polysaccharides and less proteins for many species, but not all. Understanding how chemical composition change with altered C availability is a first step towards understanding their role in organic matter accumulation and decomposition.

Vegetation, topography, and soil depth drive microbial community structure in two Swedish grasslands.

Soil microbial diversity and community composition are shaped by various factors linked to land management, topographic position, and vegetation. To study the effects of these drivers, we characterized fungal and bacterial communities from bulk soil at four soil depths ranging from the surface to below the rooting zone of two Swedish grasslands with differing land-use histories, each including both an upper and a lower catenary position. We hypothesized that differences in plant species richness and plant functional group composition between the four study sites would drive the variation in soil microbial community composition and correlate with microbial diversity, and that microbial biomass and diversity would decrease with soil depth following a decline in resource availability. While vegetation was identified as the main driver of microbial community composition, the explained variation was significantly higher for bacteria than for fungi, and the communities differed more between grasslands than between catenary positions. Microbial biomass derived from DNA abundance decreased with depth, but diversity remained relatively stable, indicating diverse microbial communities even below the rooting zone. Finally, plant-microbial diversity correlations were significant only for specific plant and fungal functional groups, emphasizing the importance of functional interactions over general species richness.

Temporal complementarity between roots and mycorrhizal fungi drives wheat nitrogen use efficiency.

Improving nitrogen (N) use efficiency (NUE) to reduce the application of N fertilisers in a way that benefits the environment and reduces farmers' costs is an ongoing objective for sustainable wheat production. However, whether and how arbuscular mycorrhizal fungi (AMF) affect NUE in wheat is still not well explored. Three independent but complementary experiments were conducted to decipher the contribution of roots and AMF to the N uptake and utilisation efficiency in wheat. We show a temporal complementarity pattern between roots and AMF in shaping NUE of wheat. Pre-anthesis N uptake efficiency mainly depends on root functional traits, but the efficiency to utilise the N taken up during pre-anthesis for producing grains (EN,g ) is strongly affected by AMF, which might increase the uptake of phosphorus and thereby improve photosynthetic carbon assimilation. Root association with AMF reduced the N remobilisation efficiency in varieties with high EN,g ; whilst the overall grain N concentration increased, due to a large improvement in post-anthesis N uptake supported by AMF and/or other microbes. The findings provide evidence for the importance of managing AMF in agroecosystems, and an opportunity to tackle the contradiction between maximising grain yield and protein concentration in wheat breeding.

Survival and growth of saprotrophic and mycorrhizal fungi in recalcitrant amine, amide and ammonium containing media.

The elimination of hazardous compounds in chemical wastes can be a complex and technically demanding task. In the search for environmental-friendly technologies, fungal mediated remediation and removal procedures are of concern. In this study, we investigated whether there are fungal species that can survive and grow on solely amine-containing compounds. One compound containing a primary amine group; 2-diethylaminoethanol, one compound with a primary amide group; 2,6-dichlorobenzamide (BAM), and a third compound containing a quaternary ammonium group; N3-trimethyl(2-oxiranyl)methanaminium chloride, were selected. The choice of these compounds was motivated by their excessive use in large scale manufacturing of protein separation media (2-diethylaminoethanol and the quaternary amine). 2,6-dichlorobenzamide, the degradation product of the herbicide 2,6-dichlorobenzonitrile (dichlobenil), was chosen since it is an extremely recalcitrant compound. Utilising part of the large fungal diversity in Northern European forests, a screening study using 48 fungal isolates from 42 fungal species, including saprotrophic and mycorrhizal fungi, was performed to test for growth responses to the chosen compounds. The ericoid (ERM) mycorrhizal fungus Rhizoscyphus ericae showed the best overall growth on 2-diethylaminoethanol and BAM in the 1-20 g L-1 concentration range, with a 35-fold and 4.5-fold increase in biomass, respectively. For N3-trimethyl(2-oxiranyl)methanaminium chloride, the peak growth occurred at 1 g L-1. In a second experiment, including three of the most promising fungi (Laccaria laccata, Hygrophorus camarophyllus and Rhizoscyphus ericae) from the screening experiment, a simulated process water containing 1.9% (w/v) 2-diethylaminoethanol and 0.8% (w/v) N3-trimethyl(2-oxiranyl)methanaminium chloride was used. Laccaria laccata showed the best biomass increase (380%) relative to a control, while the accumulation for Rhizoscyphus ericae and Hygrophorus camarophyllus were 292% and 136% respectively, indicating that mycorrhizal fungi can use amine- and amide-containing substrates as nutrients. These results show the potential of certain fungal species to be used in alternative green wastewater treatment procedures.

Site-Dependent Relationships Between Fungal Community Composition, Plant Genotypic Diversity and Environmental Drivers in a Salix Biomass System.

Soil fungi are strongly affected by plant species or genotypes since plants modify their surrounding environment, but the effects of plant genotype diversity on fungal diversity and function have not been extensively studied. The interactive responses of fungal community composition to plant genotypic diversity and environmental drivers were investigated in Salix biomass systems, posing questions about: (1) How fungal diversity varies as a function of plant genotype diversity; (2) If plant genotype identity is a strong driver of fungal community composition also in plant mixtures; (3) How the fungal communities change through time (seasonally and interannually)?; and (4) Will the proportion of ECM fungi increase over the rotation? Soil samples were collected over 4 years, starting preplanting from two Salix field trials, including four genotypes with contrasting phenology and functional traits, and genotypes were grown in all possible combinations (four genotypes in Uppsala, Sweden, two in Rostock, Germany). Fungal communities were identified, using Pacific Biosciences sequencing of fungal ITS2 amplicons. We found some site-dependent relationships between fungal community composition and genotype or diversity level, and site accounted for the largest part of the variation in fungal community composition. Rostock had a more homogenous community structure, with significant effects of genotype, diversity level, and the presence of one genotype ("Loden") on fungal community composition. Soil properties and plant and litter traits contributed to explaining the variation in fungal species composition. The within-season variation in composition was of a similar magnitude to the year-to-year variation. The proportion of ECM fungi increased over time irrespective of plant genotype diversity, and, in Uppsala, the 4-mixture showed a weaker response than other combinations. Species richness was generally higher in Uppsala compared with that in Rostock and increased over time, but did not increase with plant genotype diversity. This significant site-specificity underlines the need for consideration of diverse sites to draw general conclusions of temporal variations and functioning of fungal communities. A significant increase in ECM colonization of soil under the pioneer tree Salix on agricultural soils was evident and points to changed litter decomposition and soil carbon dynamics during Salix growth.

Variation in hyphal production rather than turnover regulates standing fungal biomass in temperate hardwood forests.

Soil fungi link above- and belowground carbon (C) fluxes through their interactions with plants and contribute to C and nutrient dynamics through the production, turnover, and activity of fungal hyphae. Despite their importance to ecosystem processes, estimates of hyphal production and turnover rates are relatively uncommon, especially in temperate hardwood forests. We sequentially harvested hyphal ingrowth bags to quantify the rates of Dikarya (Ascomycota and Basidiomycota) hyphal production and turnover in three hardwood forests in the Midwestern United States, where plots differed in their abundance of arbuscular (AM)- vs. ectomycorrhizal (ECM)-associated trees. Hyphal production rates increased linearly with the percentage of ECM trees and annual production rates were 66% higher in ECM- than AM-dominated plots. Hyphal turnover rates did not differ across the mycorrhizal gradient (plots varying in their abundance of AM vs. ECM trees), suggesting that the greater fungal biomass in ECM-dominated plots relates to greater fungal production rather than slower fungal turnover. Differences in hyphal production across the gradient aligned with distinctly different fungal communities and activities. As ECM trees increased in dominance, fungi inside ingrowth bags produced more extracellular enzymes involved in degrading nitrogen (N)-bearing relative to C-bearing compounds, suggesting greater fungal (and possibly plant) N demand in ECM-dominated soils. Collectively, our results demonstrate that shifts in temperate tree species composition that result in changes in the dominant type of mycorrhizal association may have strong impacts on Dikarya hyphal production, fungal community composition and extracellular enzyme activity, with important consequences for soil C and N cycling.

Mixture of Salix Genotypes Promotes Root Colonization With Dark Septate Endophytes and Changes P Cycling in the Mycorrhizosphere.

The roots of Salix spp. can be colonized by two types of mycorrhizal fungi (ectomycorrhizal and arbuscular) and furthermore by dark-septate endophytes. The fungal root colonization is affected by the plant genotype, soil properties and their interactions. However, the impact of host diversity accomplished by mixing different Salix genotypes within the site on root-associated fungi and P-mobilization in the field is not known. It can be hypothesized that mixing of genotypes with strong eco-physiological differences changes the diversity and abundance of root-associated fungi and P-mobilization in the mycorrhizosphere based on different root characteristics. To test this hypothesis, we have studied the mixture of two fundamentally eco-physiologically different Salix genotypes (S. dasyclados cv. 'Loden' and S. schwerinii × S. viminalis cv. 'Tora') compared to plots with pure genotypes in a randomized block design in a field experiment in Northern Germany. We assessed the abundance of mycorrhizal colonization, fungal diversity, fine root density in the soil and activities of hydrolytic enzymes involved in P-mobilization in the mycorrhizosphere in autumn and following spring after three vegetation periods. Mycorrhizal and endophytic diversity was low under all Salix treatments with Laccaria tortilis being the dominating ectomyorrhizal fungal species, and Cadophora and Paraphaeosphaeria spp. being the most common endophytic fungi. Interspecific root competition increased richness and root colonization by endophytic fungi (four taxa in the mixture vs. one found in the pure host genotype cultures) more than by ectomycorrhizal fungi and increased the activities of hydrolytic soil enzymes involved in the P-mineralization (acid phosphatase and ß-glucosidase) in mixed stands. The data suggest selective promotion of endophytic root colonization and changed competition for nutrients by mixture of Salix genotypes.

Two Salix Genotypes Differ in Productivity and Nitrogen Economy When Grown in Monoculture and Mixture.

Individual plant species or genotypes often differ in their demand for nutrients; to compete in a community they must be able to acquire more nutrients (i.e., uptake efficiency) and/or use them more efficiently for biomass production than their competitors. These two mechanisms are often complementary, as there are inherent trade-offs between them. In a mixed-stand, species with contrasting nutrient use patterns interact and may use their resources to increase productivity in different ways. Under contrasting nutrient availabilities, the competitive advantages conferred by either strategy may also shift, so that the interaction between resource use strategy and resource availability ultimately determines the performance of individual genotypes in mixtures. The aim was to investigate growth and nitrogen (N) use efficiency of two willow (Salix) genotypes grown in monoculture and mixture in a fertilizer contrast. We explored the hypotheses that (1) the biomass production of at least one of the involved genotypes should be greater when grown in mixture as compared to the corresponding monoculture when nutrients are the most growth-limiting factor; and (2) the N economy of individual genotypes differs when grown in mixture compared to the corresponding monoculture. The genotypes 'Tora' (Salix schwerinii ×S. viminalis) and 'Loden' (S. dasyclados), with contrasting phenology and functional traits, were grown from cuttings in a growth container experiment under two nutrient fertilization treatments (high and low) in mono- and mixed-culture for 17 weeks. Under low nutrient level, 'Tora' showed a higher biomass production (aboveground biomass, leaf area productivity) and N uptake efficiency in mixture than in monoculture, whereas 'Loden' showed the opposite pattern. In addition, 'Loden' showed higher leaf N productivity but lower N uptake efficiency than 'Tora.' The results demonstrated that the specific functional trait combinations of individual genotypes affect their response to mixture as compared to monoculture. Plants grown in mixture as opposed to monoculture may thus increase biomass and vary in their response of N use efficiency traits. However, young plants were investigated here, and as we cannot predict mixture response in mature stands, our results need to be validated at field scale.

Changes in turnover rather than production regulate biomass of ectomycorrhizal fungal mycelium across a Pinus sylvestris chronosequence.

In boreal forest soils, ectomycorrhizal fungi are fundamentally important for carbon (C) dynamics and nutrient cycling. Although their extraradical mycelium (ERM) is pivotal for processes such as soil organic matter build-up and nitrogen cycling, very little is known about its dynamics and regulation. In this study, we quantified ERM production and turnover, and examined how these two processes together regulated standing ERM biomass in seven sites forming a chronosequence of 12- to 100-yr-old managed Pinus sylvestris forests. This was done by determining ERM biomass, using ergosterol as a proxy, in sequentially harvested in-growth mesh bags and by applying mathematical models. Although ERM production declined with increasing forest age from 1.2 to 0.5 kg ha-1  d-1 , the standing biomass increased from 50 to 112 kg ha-1 . This was explained by a drastic decline in mycelial turnover from seven times to one time per year with increasing forest age, corresponding to mean residence times from 25 d up to 1 yr. Our results demonstrate that ERM turnover is the main factor regulating biomass across differently aged forest stands. Explicit inclusion of ERM parameters in forest ecosystem C models may significantly improve their capacity to predict responses of mycorrhiza-mediated processes to management and environmental changes.

Dominant mycorrhizal association of trees alters carbon and nutrient cycling by selecting for microbial groups with distinct enzyme function.

While it is well established that plants associating with arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) fungi cycle carbon (C) and nutrients in distinct ways, we have a limited understanding of whether varying abundance of ECM and AM plants in a stand can provide integrative proxies for key biogeochemical processes. We explored linkages between the relative abundance of AM and ECM trees and microbial functioning in three hardwood forests in southern Indiana, USA. Across each site's 'mycorrhizal gradient', we measured fungal biomass, fungal : bacterial (F : B) ratios, extracellular enzyme activities, soil carbon : nitrogen ratio, and soil pH over a growing season. We show that the percentage of AM or ECM trees in a plot promotes microbial communities that both reflect and determine the C to nutrient balance in soil. Soils dominated by ECM trees had higher F : B ratios and more standing fungal biomass than AM stands. Enzyme stoichiometry in ECM soils shifted to higher investment in extracellular enzymes needed for nitrogen and phosphorus acquisition than in C-acquisition enzymes, relative to AM soils. Our results suggest that knowledge of mycorrhizal dominance at the stand or landscape scale may provide a unifying framework for linking plant and microbial community dynamics, and predicting their effects on ecological function.

Phosphorus cycling in deciduous forest soil differs between stands dominated by ecto- and arbuscular mycorrhizal trees.

Although much is known about how trees and their associated microbes influence nitrogen cycling in temperate forest soils, less is known about biotic controls over phosphorus (P) cycling. Given that mycorrhizal fungi are instrumental for P acquisition and that the two dominant associations - arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) fungi - possess different strategies for acquiring P, we hypothesized that P cycling would differ in stands dominated by trees associated with AM vs ECM fungi. We quantified soil solution P, microbial biomass P, and sequentially extracted inorganic and organic P pools from May to November in plots dominated by trees forming either AM or ECM associations in south-central Indiana, USA. Overall, fungal communities in AM and ECM plots were functionally different and soils exhibited fundamental differences in P cycling. Organic forms of P were more available in ECM plots than in AM plots. Yet inorganic P decreased and organic P accumulated over the growing season in both ECM and AM plots, resulting in increasingly P-limited microbial biomass. Collectively, our results suggest that P cycling in hardwood forests is strongly influenced by biotic processes in soil and that these are driven by plant-associated fungal communities.

Suillus mycelia under elevated atmospheric CO2 support increased bacterial communities and scarce nifH gene activity in contrast to Hebeloma mycelia.

Bacterial communities associated with mycorrhizal roots are likely to respond to rising atmospheric CO(2) levels in terms of biomass, community composition and activity since they are supported by the carbon (C) flow outside the root tips, especially by exudation of low molecular weight organic compounds. We studied how general bacterial and diazotrophic communities associated with ectomycorrhizal (ECM) fungi respond to different belowground C supply conditions, mediated by elevated atmospheric CO(2) concentration under nitrogen (N) limited conditions. Microcosm systems were constructed using forest soil and Scots pine seedlings, which were either pre-inoculated with one of the ECM fungal species Hebeloma velutipes or Suillus variegatus, or non-inoculated. These fungal species differ in C allocation and exudation patterns. Seedlings were maintained under ambient (380 ppm) or elevated (700 ppm) CO(2) levels for 6 months. Quantitative polymerase chain reaction (PCR) showed a significant increase in 16S rRNA gene copy numbers for Suillus-inoculated microcosms under elevated CO(2) compared to ambient CO(2). The copy numbers of the nitrogenase reductase (nifH) gene were under the detection limit in all samples regardless the CO(2) treatments. Denaturing gradient gel electrophoresis analysis of PCR-amplified nifH genes revealed simple and consistent communities in all samples throughout the incubation period. A nested reverse transcription PCR approach revealed that expression of nifH genes were detected in some microcosms. Our findings suggest that the effect of mycorrhizal fungi on soil bacteria may vary depending on C supply and fungal species.

Correspondence of ectomycorrhizal diversity and colonisation of willows (Salix spp.) grown in short rotation coppice on arable sites and adjacent natural stands.

Willows (Salix spp.) are mycorrhizal tree species sometimes cultivated as short rotation coppice (SRC) on arable sites for energy purposes; they are also among the earliest plants colonising primary successional sites in natural stands. The objective of this study was to analyse the degree of colonisation and diversity of ectomycorrhizal (EM) communities on willows grown as SRC in arable soils and their adjacent natural or naturalized stands. Arable sites usually lack ectomycorrhizal host plants before the establishment of SRC, and adjacent natural or naturalized willow stands were hypothesized to be a leading source of ectomycorrhizal inoculum for the SRC. Three test sites including SRC stands (Salix viminalis, Salix dasyclados, and Salix schwerinii) and adjacent natural or naturalized (Salix caprea, Salix fragilis, and Salix × mollissima) stands in central Sweden were investigated on EM colonisation and morphotypes, and the fungal partners of 36 of the total 49 EM fungi morphotypes were identified using molecular tools. The frequency of mycorrhizas in the natural/naturalized stands was higher (two sites) or lower (one site) than in the corresponding cultivated stands. Correspondence analysis revealed that some EM taxa (e.g. Agaricales) were mostly associated with cultivated willows, while others (e.g. Thelephorales) were mostly found in natural/naturalized stands. In conclusion, we found strong effects of sites and willow genotype on EM fungi formation, but poor correspondence between the EM fungi abundance and diversity in SRC and their adjacent natural/naturalized stands. The underlying mechanism might be selective promotion of some EM fungi species by more effective spore dispersal.

Elevated CO and nitrogen influence exudation of soluble organic compounds by ectomycorrhizal root systems.

Root and mycelial exudation contributes significantly to soil carbon (C) fluxes, and is likely to be altered by an elevated atmospheric carbon dioxide (CO(2)) concentration and nitrogen (N) deposition. We quantified soluble, low-molecular-weight (LMW) organic compounds exuded by ectomycorrhizal plants grown under ambient (360 p.p.m.) or elevated (710 p.p.m.) CO(2) concentrations and with different N sources. Scots pine seedlings, colonized by one of five different ectomycorrhizal or nonmycorrhizal fungi, received 70 muM N, either as NH(4)Cl or as alanine, in a liquid growth medium. Exudation of LMW organic acids (LMWOAs), dissolved monosaccharides and total dissolved organic carbon were determined. Both N and CO(2) had a significant impact on exudation, especially of LMWOAs. Exudation of LMWOAs was negatively affected by inorganic N and decreased by 30-85% compared with the organic N treatment, irrespective of the CO(2) treatment. Elevated CO(2) had a clear impact on the production of individual LMWOAs, although with very contrasting effects depending on which N source was supplied.

Ectomycorrhizal fungi in culture respond differently to increased carbon availability.

Carbon (C) availability to ectomycorrhizal fungi is likely to increase at elevated atmospheric CO(2). To determine whether there are any broad patterns in species' responses that relate to their ecology, we compared growth, respiration, N uptake and C exudation of 17 fungal isolates in liquid culture. As a surrogate for increased C availability we used three different C:N ratios (10:1, 20:1 and 40:1), moving from conditions of C limitation to conditions of N limitation. Responses were species-specific, and suilloid fungi were the most responsive in terms of growth and respiration. In contrast, a group of eight isolates showed no growth increase above C:N 20:1. This inability to respond was not due to N limitation, although there were marked differences in N uptake between isolates. At higher C availability isolates generally became more efficient in converting C into biomass. Six isolates showed net release of exudates into the culture medium (up to 40% of the C in biomass and respiration). We conclude that the findings were in agreement with field observations, and suggest that pure culture observations can yield ecologically relevant information on how ectomycorrhizal fungi may respond under conditions of elevated CO(2).

Mycelial production, spread and root colonisation by the ectomycorrhizal fungi Hebeloma crustuliniforme and Paxillus involutus under elevated atmospheric CO2.

Effects of elevated atmospheric carbon dioxide (CO2) levels on the production and spread of ectomycorrhizal fungal mycelium from colonised Scots pine roots were investigated. Pinus sylvestris (L.) Karst. seedlings inoculated with either Hebeloma crustuliniforme (Bull:Fr.) Quel. or Paxillus involutus (Fr.) Fr. were grown at either ambient (350 ppm) or elevated (700 ppm) levels of CO2. Mycelial production was measured after 6 weeks in pots, and mycelial spread from inoculated seedlings was studied after 4 months growth in perlite in shallow boxes containing uncolonised bait seedlings. Plant and fungal biomass were analysed, as well as carbon and nitrogen content of seedling shoots. Mycelial biomass production by H. crustuliniforme was significantly greater under elevated CO2 (up to a 3-fold increase was observed). Significantly lower concentrations and total amounts of N were found in plants exposed to elevated CO2.

Albatrellus citrinus sp. nov., connected to Picea abies on lime rich soils.

Field observations indicated that a morphotype of Albatrellus subrubescens seemed connected to Picea abies and lime rich soils, while the original morphotype seemed connected with Pinus sylvestris and indifferent to lime. We conducted a molecular study (ITS sequencing of 22 Albatrellus specimens) to test the hypothesis that we in fact had discovered a new species. Our results confirmed the hypothesis, i.e. the Picea taxon (604 bp ITS) had little intraspecific variation in spite of 1600 km distance between samples, but compared with the Pinus taxon (598 bp ITS) the sequence difference was constantly 5.6% regardless of close distance (75 km). We describe the new species A. citrinus, which apart from ecology and ITS sequence, is different from A. subrubescens by a distinct yellowing with age, lack of dark spots of the cap, a mild taste and somewhat narrower spores. A. citrinus seems to be more related to A. ovinus than to A. subrubescens, and A. syringae may not even be a true Albatrellus.

Species level patterns in 13 C and 15 N abundance of ectomycorrhizal and saprotrophic fungal sporocarps.

• The natural abundance of 13 C (δ13 C) and 15 N (δ15 N) of saprotrophic and ectomycorrhizal (ECM) fungi has been investigated on a number of occasions, but the significance of observed differences within and between the two trophic groups remains unclear. • Here, we examine the influence of taxonomy, site, host and time upon isotopic data from 135 fungal species collected at two forest sites in Sweden. • Mean δ13 C and δ15 N values differed significantly between ECM and saprotrophic fungi, with only a small degree of overlap even at the species level. Among ECM fungi, intraspecific variation in δ15 N was low compared with interspecific and intergeneric variation. Significant variation due to site, year and host association was found. • At broad scales a number of factors clearly influence δ13 C and δ15 N values making interpretation problematic. We suggest that values are essentially site-specific within the two trophic groups, but that species-level patterns exist potentially reflecting ecophysiological attributes of species. The species is therefore highlighted as the taxonomic level at which most information may be obtained from fungal δ13 C and δ15 N data.

Elevated atmospheric CO2 alters root symbiont community structure in forest trees.

• Changes in below-ground ectomycorrhizal (ECM) community structure in response to elevated CO2 and balanced nutrient addition were investigated in a 37-yr-old Picea abies forest. • Trees in whole-tree chambers were exposed to factorial combinations of ambient/elevated CO2 (700 ppm) and fertilization (+/-). ECM fungal community structure was determined in 1997 and 2000 using a combination of morphotyping and molecular analyses. Samples were taken both from chambers and from reference trees receiving the same fertilization treatments but without chambers. • Significant effects on ECM community structure were found in response to elevated CO2 . Neither elevated CO2 nor fertilization altered species richness; however, there was considerable variation among samples, which may have masked treatment effects on individual species. After 3 yr, the effects of elevated CO2 on community composition were of the same magnitude as those seen after 15 yr of fertilization treatment. • Our results show that increasing atmospheric CO2 concentrations affect the community structure of root symbionts colonizing forest trees. The potential effects of altered ECM community structure on allocation and turnover of carbon and nutrients within forest ecosystems are discussed.

Effects of continuous optimal fertilization on belowground ectomycorrhizal community structure in a Norway spruce forest.

Studies of effects of fertilizer treatment on ectomycorrhizal fungal community structure have predominantly been based on large, single additions of nitrogen. Studies involving chronic additions of nutrients in combination with irrigation are much less common. We used morphotyping to study effects of balanced additions of a nutrient solution on ectomycorrhizal fungal community structure in a 36-year-old stand of Picea abies (L.) Karst. Despite high variability among individual samples, principal components analysis revealed a clear shift in community structure in response to fertilization. Irrigated plots receiving only water did not differ significantly from untreated control plots. Mycorrhizal root tips colonized by Cenococcum geophilum Fr. were significantly more common in fertilized plots than in control plots. Possible responses by other ectomycorrhizal species were masked by high variability. Over sixty morphotypes were distinguished, but there was no measurable effect of either fertilizer or irrigation treatment on morphotype richness or total number of root tips.