Phylogenetic and functional traits of ectomycorrhizal assemblages in top soil from different biogeographic regions and forest types.

Ectomycorrhizal (EM) fungal taxonomic, phylogenetic, and trait diversity (exploration types) were analyzed in beech and conifer forests along a north-to-south gradient in three biogeographic regions in Germany. The taxonomic community structures of the ectomycorrhizal assemblages in top soil were influenced by stand density and forest type, by biogeographic environmental factors (soil physical properties, temperature, and precipitation), and by nitrogen forms (amino acids, ammonium, and nitrate). While α-diversity did not differ between forest types, ß-diversity increased, leading to higher γ-diversity on the landscape level when both forest types were present. The highest taxonomic diversity of EM was found in forests in cool, moist climate on clay and silty soils and the lowest in the forests in warm, dry climate on sandy soils. In the region with higher taxonomic diversity, phylogenetic clustering was found, but not trait clustering. In the warm region, trait clustering occurred despite neutral phylogenetic effects. These results suggest that different forest types and favorable environmental conditions in forests promote high EM species richness in top soil presumably with both high functional diversity and phylogenetic redundancy, while stressful environmental conditions lead to lower species richness and functional redundancy.

Ectomycorrhizal identification in environmental samples of tree roots by Fourier-transform infrared (FTIR) spectroscopy.

Roots of forest trees are associated with various ectomycorrhizal (ECM) fungal species that are involved in nutrient exchange between host plant and the soil compartment. The identification of ECM fungi in small environmental samples is difficult. The present study tested the feasibility of attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy followed by hierarchical cluster analysis (HCA) to discriminate in situ collected ECM fungal species. Root tips colonized by distinct ECM fungal species, i.e., Amanita rubescens, Cenococcum geophilum, Lactarius subdulcis, Russula ochroleuca, and Xerocomus pruinatus were collected in mono-specific beech (Fagus sylvatica) and mixed deciduous forests in different geographic areas to investigate the environmental variability of the ECM FTIR signatures. A clear HCA discrimination was obtained for ECM fungal species independent of individual provenance. Environmental variability neither limited the discrimination between fungal species nor provided sufficient resolution to discern species sub-clusters for different sites. However, the de-convoluted FTIR spectra contained site-related spectral information for fungi with wide nutrient ranges, but not for Lactarius subdulcis, a fungus residing only in the litter layer. Specific markers for distinct ECM were identified in spectral regions associated with carbohydrates (i.e., mannans), lipids, and secondary protein structures. The present results support that FTIR spectroscopy coupled with multivariate analysis is a reliable and fast method to identify ECM fungal species in minute environmental samples. Moreover, our data suggest that the FTIR spectral signatures contain information on physiological and functional traits of ECM fungi.

Spatial patterns of ectomycorrhizal assemblages in a monospecific forest in relation to host tree genotype.

Ectomycorrhizas (EcM) are important for soil exploration and thereby may shape belowground interactions of roots. We investigated the composition and spatial structures of EcM assemblages in relation to host genotype in an old-growth, monospecific beech (Fagus sylvatica) forest. We hypothesized that neighboring roots of different beech individuals are colonized by similar EcM assemblages if host genotype had no influence on the fungal colonization and that the similarity would decrease with increasing distance of the sampling points. The alternative was that the EcM species showed preferences for distinct beech genotypes resulting in intraspecific variation of EcM-host assemblages. EcM species identities, abundance and exploration type as well as the genotypes of the colonized roots were determined in each sampling unit of a 1 L soil core (r = 0.04 m, depth 0.2 m). The Morisita-Horn similarity indices (MHSI) based on EcM species abundance and multiple community comparisons were calculated. No pronounced variation of MHSI with increasing distances of the sampling points within a plot was found, but variations between plots. Very high similarities and no between plot variation were found for MHSI based on EcM exploration types suggesting homogenous soil foraging in this ecosystem. The EcM community on different root genotypes in the same soil core exhibited high similarity, whereas the EcM communities on the root of the same tree genotype in different soil cores were significantly dissimilar. This finding suggests that spatial structuring of EcM assemblages occurs within the root system of an individual. This may constitute a novel, yet unknown mechanism ensuring colonization by a diverse EcM community of the roots of a given host individual.

General relationships between abiotic soil properties and soil biota across spatial scales and different land-use types.

Very few principles have been unraveled that explain the relationship between soil properties and soil biota across large spatial scales and different land-use types. Here, we seek these general relationships using data from 52 differently managed grassland and forest soils in three study regions spanning a latitudinal gradient in Germany. We hypothesize that, after extraction of variation that is explained by location and land-use type, soil properties still explain significant proportions of variation in the abundance and diversity of soil biota. If the relationships between predictors and soil organisms were analyzed individually for each predictor group, soil properties explained the highest amount of variation in soil biota abundance and diversity, followed by land-use type and sampling location. After extraction of variation that originated from location or land-use, abiotic soil properties explained significant amounts of variation in fungal, meso- and macrofauna, but not in yeast or bacterial biomass or diversity. Nitrate or nitrogen concentration and fungal biomass were positively related, but nitrate concentration was negatively related to the abundances of Collembola and mites and to the myriapod species richness across a range of forest and grassland soils. The species richness of earthworms was positively correlated with clay content of soils independent of sample location and land-use type. Our study indicates that after accounting for heterogeneity resulting from large scale differences among sampling locations and land-use types, soil properties still explain significant proportions of variation in fungal and soil fauna abundance or diversity. However, soil biota was also related to processes that act at larger spatial scales and bacteria or soil yeasts only showed weak relationships to soil properties. We therefore argue that more general relationships between soil properties and soil biota can only be derived from future studies that consider larger spatial scales and different land-use types.

Ectomycorrhizal fungal diversity, tree diversity and root nutrient relations in a mixed Central European forest.

Knowledge is limited about whether root nutrient concentrations are affected by mixtures of tree species and interspecific root competition. The goal of this field study was to investigate root nutrient element concentrations in relation to root and ectomycorrhizal (EM) diversity in six different mixtures of beech (Fagus sylvatica), ash (Fraxinus excelsior) and lime (Tilia sp.) in an old-growth, undisturbed forest ecosystem. Root biomass and nutrient concentrations per tree taxon as well as the abundance and identity of all EM fungi were determined in soil cores of a volume of 1 L (r=40 mm, depth=200 mm). Stand-level nutrient concentrations in overall root biomass and H' (Shannon-Wiener diversity) were obtained by pooling the data per stand. At stand level, Shannon H' for roots and aboveground tree species abundance were correlated. H' for roots and EM fungi were not correlated because of the contribution of ash roots that form only arbuscular mycorrhizal but no EM associations. Nutrient element concentrations in roots showed taxon-related differences and increased in the following order: beech ≤ lime < ash with the exception of calcium (Ca), which was lower in ash. Stand-level concentrations of Ca, magnesium, potassium and sulfur in roots increased with increasing tree diversity because of two effects: increasing contribution of ash roots to the mixture and increasing Ca accumulation in beech roots with increasing root diversity. On a small scale, increasing root diversity, but not EM diversity, was correlated with decreasing P concentrations in beech roots pointing to interspecific tree competition. Nitrogen (N) concentrations of beech roots were unaltered in relation to root and EM diversity. Opposing behavior was observed for lime and ash: the N concentrations in lime roots increased, whereas those in ash roots decreased with increasing EM diversity in a given soil volume. This suggests that EM diversity facilitates N acquisition of lime roots at the expense of non-EM ash.

Host preferences and differential contributions of deciduous tree species shape mycorrhizal species richness in a mixed Central European forest.

Mycorrhizal species richness and host ranges were investigated in mixed deciduous stands composed of Fagus sylvatica, Tilia spp., Carpinus betulus, Acer spp., and Fraxinus excelsior. Acer and Fraxinus were colonized by arbuscular mycorrhizas and contributed 5% to total stand mycorrhizal fungal species richness. Tilia hosted similar and Carpinus half the number of ectomycorrhizal (EM) fungal taxa compared with Fagus (75 putative taxa). The relative abundance of the host tree the EM fungal richness decreased in the order Fagus > Tilia >> Carpinus. After correction for similar sampling intensities, EM fungal species richness of Carpinus was still about 30-40% lower than that of Fagus and Tilia. About 10% of the mycorrhizal species were shared among the EM forming trees; 29% were associated with two host tree species and 61% with only one of the hosts. The latter group consisted mainly of rare EM fungal species colonizing about 20% of the root tips and included known specialists but also putative non-host associations such as conifer or shrub mycorrhizas. Our data indicate that EM fungal species richness was associated with tree identity and suggest that Fagus secures EM fungal diversity in an ecosystem since it shared more common EM fungi with Tilia and Carpinus than the latter two among each other.

Beech carbon productivity as driver of ectomycorrhizal abundance and diversity.

We tested the hypothesis that carbon productivity of beech (Fagus sylvatica) controls ectomycorrhizal colonization, diversity and community structures. Carbon productivity was limited by long-term shading or by girdling. The trees were grown in compost soil to avoid nutrient deficiencies. Despite severe limitation in photosynthesis and biomass production by shading, the concentrations of carbohydrates in roots were unaffected by the light level. Shade-acclimated plants were only 10% and sun-acclimated plants were 74% colonized by ectomycorrhiza. EM diversity was higher on roots with high than at roots with low mycorrhizal colonization. Evenness was unaffected by any treatment. Low mycorrhizal colonization had no negative effects on plant mineral nutrition. In girdled plants mycorrhizal colonization and diversity were retained although (14)C-leaf feeding showed almost complete disruption of carbon transport from leaves to roots. Carbohydrate storage pools in roots decreased upon girdling. Our results show that plant carbon productivity was the reason for and not the result of high ectomycorrhizal diversity. We suggest that ectomycorrhiza can be supplied by two carbon routes: recent photosynthate and stored carbohydrates. Storage pools may be important for ectomycorrhizal survival when photoassimilates were unavailable, probably feeding preferentially less carbon demanding EM species as shifts in community composition were found.