Contrasting continental patterns of adaptive population divergence in the holarctic ectomycorrhizal fungus Boletus edulis.

In the hyperdiverse fungi, the process of speciation is virtually unknown, including for the > 20 000 species of ectomycorrhizal mutualists. To understand this process, we investigated patterns of genome-wide differentiation in the ectomycorrhizal porcini mushroom, Boletus edulis, a globally distributed species complex with broad ecological amplitude. By whole-genome sequencing 160 individuals from across the Northern Hemisphere, we genotyped 792 923 single nucleotide polymorphisms to characterize patterns of genome-wide differentiation and to identify the adaptive processes shaping global population structure. We show that B. edulis exhibits contrasting patterns of genomic divergence between continents, with multiple lineages present across North America, while a single lineage dominates Europe. These geographical lineages are inferred to have diverged 1.62-2.66 million years ago, during a period of climatic upheaval and the onset of glaciation in the Pliocene-Pleistocene boundary. High levels of genomic differentiation were observed among lineages despite evidence of substantial and ongoing introgression. Genome scans, demographic inference, and ecological niche models suggest that genomic differentiation is maintained by environmental adaptation, not physical isolation. Our study uncovers striking patterns of genome-wide differentiation on a global scale and emphasizes the importance of local adaptation and ecologically mediated divergence, rather than prezygotic barriers such as allopatry or genomic incompatibility, in fungal population differentiation.

Re-examination of the Southern Hemisphere truffle genus Amylascus (Pezizaceae, Ascomycota) and characterization of the sister genus Nothoamylascus gen. nov.

Amylascus is a genus of ectomycorrhizal truffles within Pezizaceae that is known from Australia and contains only two described species, A. herbertianus and A. tasmanicus. Species of Amylascus are closely related to truffles (Pachyphlodes, Luteoamylascus) and cup fungi (Plicariella) from the Northern Hemisphere. Here we reevaluate the species diversity of Amylascus and related taxa from southern South America and Australia based on new morphological and molecular data. We identify previously undocumented diversity and morphological variability in ascospore color, ascospore ornamentation, hymenial construction, epithecium structure and the amyloid reaction of the ascus in Melzer's reagent. We redescribe two Amylascus species from Australia and describe seven new Amylascus species, five from South America and two from Australia. This is the first report of Amylascus species from South America. We also describe the new South American genus Nothoamylascus as sister lineage to the Pachyphlodes-Amylascus-Luteoamylascus clade (including Amylascus, Luteoamylascus, Pachyphlodes, and Plicariella). We obtained ITS sequences of mitotic spore mats from Nothoamylascus erubescens gen. & sp. nov. and four of the seven newly described Amylascus species, providing the first evidence of mitotic spore mats in Amylascus. Additional ITS sequences from mitotic spore mats reveal the presence of nine additional undescribed Amylascus and one Nothoamylascus species that do not correspond to any sampled ascomata. We also identify three additional undescribed Amylascus species based on environmental sequences from the feces of two grounddwelling bird species from Chile, Scelorchilus rubecula and Pteroptochos tarnii. Our results indicate that ascomata from Amylascus and Nothoamylascus species are rarely collected, but molecular data from ectomycorrhizal roots and mitotic spore mats indicate that these species are probably common and widespread in southern South America. Finally, we present a time-calibrated phylogeny that is consistent with a late Gondwanan distribution. The time since the most recent common ancestor of: 1) the family Pezizaceae had a mean of 276 Ma (217-337 HPD); 2) the Amylascus-Pachyphlodes-Nothoamylascus-Luteoamylascus clade had a mean of 79 Ma (60-100 HPD); and 3) the Amylascus-Pachyphlodes clade had a mean of 50 Ma (38-62 HPD). The crown age of Pachyphlodes had a mean of 39 Ma (25-42 HPD) and Amylascus had a mean age of 28 Ma (20-37 HPD), falling near the Eocene-Oligocene boundary and the onset of the Antarctic glaciation (c. 35 Ma). Citation: Healy RA, Truong C, Castellano MA, et al. 2023. Re-examination of the Southern Hemisphere truffle genus Amylascus (Pezizaceae, Ascomycota) and characterization of the sister genus Nothoamylascus gen. nov. Persoonia 51: 125-151. doi: 10.3767/persoonia.2023.51.03.

Microbial community shifts correspond with suppression of decomposition 25 years after liming of acidic forest soils.

Microbial community structure and function regularly covary with soil pH, yet effects of these interactions on soil carbon are rarely tested experimentally within natural ecosystems. We investigated the enduring (25 year) impacts of liming on microbial community structure and decomposition at an acidic northern hardwood forest, where experimental liming increased pH one unit and surprisingly doubled the organic carbon stocks of the forest floor. We show that this increase in carbon storage corresponded with restructuring of the bacterial and fungal communities that drive decomposition. In the Oe horizon, liming reduced the activities of five extracellular enzymes that mediate decomposition, while the Oa horizon showed an especially large (64%) reduction in the activity of a sixth, peroxidase, which is an oxidative enzyme central to lignocellulose degradation. Decreased enzyme activities corresponded with loss of microbial taxa important for lignocellulose decay, including large reductions in the dominant ectomycorrhizal genera Russula and Cenococcum, saprotrophic and wood decaying fungi, and Actinobacteria (Thermomonosporaceae). These results demonstrate the importance of pH as a dominant regulator of microbial community structure and illustrate how changes to this structure can produce large, otherwise unexpected increases in carbon storage in forest soils.

Bacteria could help ectomycorrhizae establishment under climate variations.

Rhizosphere microbiome is one of the main sources of plant protection against drought. Beneficial symbiotic microorganisms, such as ectomycorrhizal fungi (ECMF) and mycorrhiza helper bacteria (MHB), interact with each other for increasing or maintaining host plant fitness. This mutual support benefits all three partners and comprises a natural system for drought acclimation in plants. Cork oak (Quercus suber L.) tolerance to drought scenarios is widely known, but adaptation to climate changes has been a challenge for forest sustainability protection. In this work, ECMF and MHB communities from cork oak forests were cross-linked and correlated with climates. Cenococcum, Russula and Tuber were the most abundant ECMF capable of interacting with MHB (ECMF~MHB) genera in cork oak stands, while Bacillus, Burkholderia and Streptomyces were the most conspicuous MHB. Integrating all microbial data, two consortia Lactarius/Bacillaceae and Russula/Burkholderaceae have singled out but revealed a negative interaction with each other. Russula/Burkholderaceae might have an important role for cork oak forest sustainability in arid environments, which will be complemented by the lower drought adaptation of competitive Lactarius/Bacillaceae. These microbial consortia could play an essential role on cork oak forest resilience to upcoming climatic changes.

A new edible Rhizopogon species from Southwest China, and its mycorrhizal synthesis with two native pines.

A new Rhizopogon species associated with Pinus was discovered at local wild mushroom markets and Pinus armandii forests from March to July in Southwest China where it is considered a delicacy. Based on morphological and molecular phylogenetic analyses, the collections were described as Rhizopogon songmaodan sp. nov. belonging to the subgenus Versicolores. The new species described here increases the current number of Rhizopogon species known in China to ten. R. songmaodan establishes ectomycorrhizal associations with P. armandii which was confirmed by comparing rDNA ITS sequences from basidiomata and ectomycorrhizal root tips. Mycorrhizal synthesis via spore inoculation between R. songmaodan and two native pine species, Pinus armandii and P. yunnanensis was successfully carried out in a greenhouse study. The ease of R. songmaodan inoculation onto pine species, and the high market demand of its sporocarps, could make R. songmaodan a good candidate for cultivation in Southwest China.

Laccaria bicolor Mobilizes both Labile Aluminum and Inorganic Phosphate in Rhizosphere Soil of Pinus massoniana Seedlings Field Grown in a Yellow Acidic Soil.

Plant growth is often limited by highly activated aluminum (Al) and low available phosphorus (P) in acidic soil. Ectomycorrhizal (ECM) fungi can improve their host plants' Al tolerance by increasing P availability while decreasing Al activity in vitro or in hydroponic or sand culture systems. However, the effect of ECM fungi on inorganic P (IP) and labile Al in acidic soil in the field, particularly in conjunction with Al treatment, remains poorly understood. The present study aimed to determine the influence of ECM fungal association on the mobilization of IP and labile Al in rhizosphere soil of host plants grown in the field with external Al treatment and the underlying nutritional mechanism in plant Al tolerance. To do so, 4-week-old Pinus massoniana seedlings were inoculated with three ECM isolates (Laccaria bicolor 270, L. bicolor S238A, and L. bicolor S238N) and grown in a Haplic Alisol field with or without Al treatment for 12 weeks. Results showed that L. bicolor association enhanced the available P depletion and facilitated the mobilization of IP and labile Al, in turn improving the capacity of host plant to use Al-bound P, Ca-bound P, and occluded P, particularly when P. massoniana seedlings were inoculated with L. bicolor S238A. Inoculation with L. bicolor isolates also enhanced the solubility of labile Al and facilitated the conversion of acid-soluble Al into exchangeable Al. Our findings suggested that ECM inoculation could enhance plant Al tolerance in the field by mobilizing IP to improve the P bioavailability but not by decreasing Al activity.IMPORTANCE Here, we reveal the underlying nutritional mechanism in plant Al tolerance conferred by ectomycorrhizal (ECM)-fungus inoculation in the field and report the screening of a promising ECM isolate to assist phytoremediation and afforestation using Pinus massoniana in acidic soil in southern China. This study advances our understanding of the contribution of ECM fungi to plant-ECM-fungus symbiosis and highlights the vital role of ECM-fungus inoculation in plant Al tolerance. In addition, the results described in the present study confirm the importance of carrying out studies in the field rather than only in vitro studies. Our findings strengthen our understanding of the role of ECM-fungus association in detecting, utilizing, and transporting unavailable nutrients in the soil to enhance host plant growth and adaptability in response to adverse habitats.

Structural and Functional Dynamics of Soil Microbes following Spruce Beetle Infestation.

As the range of bark beetles expands into new forests and woodlands, the need to understand their effects on multiple trophic levels becomes increasingly important. To date, much attention has been paid to the aboveground processes affected by bark beetle infestation, with a focus on photoautotrophs and ecosystem level processes. However, indirect effects of bark beetle on belowground processes, especially the structure and function of soil microbiota remains largely a black box. Our study examined the impacts of bark beetle-induced tree mortality on soil microbial community structure and function using high-throughput sequencing of the soil bacterial and fungal communities and measurements of extracellular enzyme activities. The results suggest bark beetle infestation affected edaphic conditions through increased soil water content, pH, electrical conductivity, and carbon/nitrogen ratio and altered bulk and rhizosphere soil microbial community structure and function. Finally, increased enzymatic activity suggests heightened microbial decomposition following bark beetle infestation. With this increase in enzymatic activity, nutrients trapped in organic substrates may become accessible to seedlings and potentially alter the trajectory of forest regeneration. Our results indicate the need for incorporation of microbial processes into ecosystem level models.IMPORTANCE Belowground impacts of bark beetle infestation have not been explored as thoroughly as their aboveground counterparts. In order to accurately model impacts of bark beetle-induced tree mortality on carbon and nutrient cycling and forest regeneration, the intricacies of soil microbial communities must be examined. In this study, we investigated the structure and function of soil bacterial and fungal communities following bark beetle infestation. Our results show bark beetle infestation to impact soil conditions, as well as soil microbial community structure and function.

An ectomycorrhizal fungus alters sensitivity to jasmonate, salicylate, gibberellin, and ethylene in host roots.

The phytohormones jasmonate, gibberellin, salicylate, and ethylene regulate an interconnected reprogramming network integrating root development with plant responses against microbes. The establishment of mutualistic ectomycorrhizal symbiosis requires the suppression of plant defense responses against fungi as well as the modification of root architecture and cortical cell wall properties. Here, we investigated the contribution of phytohormones and their crosstalk to the ontogenesis of ectomycorrhizae (ECM) between grey poplar (Populus tremula x alba) roots and the fungus Laccaria bicolor. To obtain the hormonal blueprint of developing ECM, we quantified the concentrations of jasmonates, gibberellins, and salicylate via liquid chromatography-tandem mass spectrometry. Subsequently, we assessed root architecture, mycorrhizal morphology, and gene expression levels (RNA sequencing) in phytohormone-treated poplar lateral roots in the presence or absence of L. bicolor. Salicylic acid accumulated in mid-stage ECM. Exogenous phytohormone treatment affected the fungal colonization rate and/or frequency of Hartig net formation. Colonized lateral roots displayed diminished responsiveness to jasmonate but regulated some genes, implicated in defense and cell wall remodelling, that were specifically differentially expressed after jasmonate treatment. Responses to salicylate, gibberellin, and ethylene were enhanced in ECM. The dynamics of phytohormone accumulation and response suggest that jasmonate, gibberellin, salicylate, and ethylene signalling play multifaceted roles in poplar L. bicolor ectomycorrhizal development.

Gap creation alters the mode of conspecific distance-dependent seedling establishment via changes in the relative influence of pathogens and mycorrhizae.

In forest communities, conspecific density/distance dependence (CDD) is an important factor regulating diversity. It remains unknown how and the extent to which gap creation alters the mode and strength of CDD via changes in the relative importance of pathogens and mycorrhizae. Seeds of two hardwoods (i.e., Acer mono associated with arbuscular mycorrhizae [AM] and Quercus serrata associated with ectomycorrhizae [EM]) were sown reciprocally at four distances from the boundary between Acer- and Quercus-dominated forests towards forest interior in each of forest understories (FUs) and gaps. The causes of seed and seedling mortality, seedling growth and colonization of mycorrhizal fungi were investigated. In Acer, seed and seedling mortality were highest in Acer forests and gradually decreased towards the interior of Quercus forests in FU, mainly due to severe attack of soil pathogens, invertebrates, and leaf diseases. The reverse was true in gaps, due to reduction of damping-off damage caused by distance-dependent colonization of AM. In Quercus, most seeds and seedlings were eaten by vertebrates in FUs. The seedling mortality caused by leaf diseases was not high, even beneath conspecific forests with higher colonization of EM in gaps, suggesting a positive EM influence. In both species, seedling mass was greatest in conspecific forests and gradually decreased towards the interior of heterospecific forests in gaps, due to higher colonization of mycorrhizae near conspecifics. In conclusion, light conditions strongly altered the mode of CDD via changes in relative influence of pathogens and mycorrhizae, suggesting that gap creation may regulate species diversity via changes in the mode of CDD.

Persistence of Tuber melanosporum in truffle orchards in North Carolina, USA.

A survey was conducted to determine the persistence of mycorrhization by Tuber melanosporum in truffle orchards established with European and American species of oak and common hazel trees in North Carolina. The trees had reportedly been inoculated and colonized by T. melanosporum prior to planting. Root samples were collected from 95 trees among seven orchards in 2015 and roots were analyzed by morphology and quantitative PCR. Samples that tested negative for T. melanosporum or where ectomycorrhizal morphology was not observed were analyzed by sequencing to identify the mycorrhizal fungal symbiont present. The presence of T. melanosporum was detected in all seven orchards. In six orchards, T. melanosporum was detected on all trees, but in only two of fifteen trees in one orchard. Other species of Tuber including T. brennemanii, T. canaliculatum, and T. lyonii, species of Scleroderma, and members of the Pezizales were also detected by sequence analysis. Sporocarps of T. aestivum and T. brumale were found in 2017 and 2018 in separate orchards in North Carolina after the survey was conducted. Overall, results indicate that T. melanosporum has persisted in truffle orchards sampled in North Carolina. Indigenous and contaminating fungal species, including Tuber species, were also detected and present a challenge to the truffle industry in North Carolina.

Comparison of Actinomycete Community Composition on the Surface and Inside of Japanese Black Pine (Pinus thunbergii) Tree Roots Colonized by the Ectomycorrhizal Fungus Cenococcum geophilum.

Various bacteria are associated with ectomycorrhizal roots, which are symbiotic complexes formed between plant roots and fungi. Among these associated bacteria, actinomycetes have received attention for their ubiquity and diverse roles in forest ecosystems. Here, to examine the association of actinomycetes with ectomycorrhizal root tips, we compared the bacterial and actinomycete communities on the surface and inside of root tips of coastal Japanese black pine (Pinus thunbergii) colonized by the fungus Cenococcum geophilum. Next-generation sequences of 16S rDNA of bacteria communities using the Ion Torrent Personal Genome Machine showed that the number of bacterial classes in the surface of C. geophilum ECM roots was significantly higher than that in non-ECM roots. The bacterial community structure of surface, inside, and non-ECM roots was significantly discriminated each other. For an isolation method, a total of 762 and 335 actinomycete isolates were obtained from the surface and inside of the roots, respectively. In addition, the isolation ratio of actinomycetes in these root tips varied depending on the age of the tree and the season. Identification of the isolates based on partial 16S rDNA sequencing revealed that the isolates belonged to nine genera of the order Actinomycetales. On the surface of the roots, most of the isolates belonged to genus Streptomyces (90.4%); inside of the roots, most of the isolates belonged to genus Actinoallomurus (40.0%), which is a relatively new taxon. Our results suggest that actinomycetes as well as bacteria are ubiquitously associated with C. geophilum ectomycorrhizal roots of P. thunbergii, although their communities can vary either surface or inside of individual root tips.

New insights into black truffle biology: discovery of the potential connecting structure between a Tuber aestivum ascocarp and its host root.

According to isotopic labeling experiments, most of the carbon used by truffle (Tuber sp.) fruiting bodies to develop underground is provided by host trees, suggesting that trees and truffles are physically connected. However, such physical link between trees and truffle fruiting bodies has never been observed. We discovered fruiting bodies of Tuber aestivum adhering to the walls of a belowground quarry and we took advantage of this unique situation to analyze the physical structure that supported these fruiting bodies in the open air. Observation of transversal sections of the attachment structure indicated that it was organized in ducts made of gleba-like tissue and connected to a network of hyphae traveling across soil particles. Only one mating type was detected by PCR in the gleba and in the attachment structure, suggesting that these two organs are from maternal origin, leaving open the question of the location of the opposite paternal mating type.

In Planta Sporulation of Frankia spp. as a Determinant of Alder-Symbiont Interactions.

The Alnus genus forms symbiosis with the actinobacteria Frankia spp. and ectomycorrhizal fungi. Two types of Frankia lineages can be distinguished based on their ability to sporulate in planta Spore-positive (Sp+) strains are predominant on Alnus incana and Alnus viridis in highlands, while spore-negative (Sp-) strains are mainly associated with Alnus glutinosa in lowlands. Here, we investigated whether the Sp+ predominance in nodules is due to host selection of certain Frankia genotypes from soil communities or the result of the ecological history of the alder stand soil, as well as the effect of the sporulation genotype on the ectomycorrhizal (ECM) communities. Trapping experiments were conducted using A. glutinosa, A. incana, and A. viridis plantlets on 6 soils, differing in the alder species and the frequency of Sp+ nodules in the field. Higher diversity of Frankia spp. and variation in Sp+ frequencies were observed in the trapping than in the fields. Both indigenous and trapping species shape Frankia community structure in trapped nodules. Nodulation impediments were observed under several trapping conditions in Sp+ soils, supporting a narrower host range of Sp+ Frankia species. A. incana and A. viridis were able to associate equally with compatible Sp+ and Sp- strains in the greenhouse. Additionally, no host shift was observed for Alnus-specific ECM, and the sporulation genotype of Frankia spp. defined the ECM communities on the host roots. The symbiotic association is likely determined by the host range, the soil history, and the type of in plantaFrankia species. These results provide an insight into the biogeographical drivers of alder symbionts in the Holarctic region.IMPORTANCE Most Frankia-actinorhiza plant symbioses are capable of high rates of nitrogen fixation comparable to those found on legumes. Yet, our understanding of the ecology and distribution of Frankia spp. is still very limited. Several studies have focused on the distribution patterns of Frankia spp., demonstrating a combination of host and pedoclimatic parameters in their biogeography. However, very few have considered the in planta sporulation form of the strain, although it is a unique feature among all symbiotic plant-associated microbes. Compared with Sp- Frankia strains, Sp+ strains would be obligate symbionts that are highly dependent on the presence of a compatible host species and with lower efficiency in nitrogen fixation. Understanding the biogeographical drivers of Sp+ Frankia strains might help elucidate the ecological role of in planta sporulation and the extent to which this trait mediates host-partner interactions in the alder-Frankia-ECM fungal symbiosis.

Quantitative losses vs. qualitative stability of ectomycorrhizal community responses to 3 years of experimental summer drought in a beech-spruce forest.

Forest ecosystems in central Europe are predicted to face an increasing frequency and severity of summer droughts because of global climate change. European beech and Norway spruce often coexist in these forests with mostly positive effects on their growth. However, their different below-ground responses to drought may lead to differences in ectomycorrhizal (ECM) fungal community composition and functions which we examined at the individual root and ecosystem levels. We installed retractable roofs over plots in Kranzberg Forest (11°39'42″E, 48°25'12″N; 490 m a.s.l.) to impose repeated summer drought conditions and assigned zones within each plot where trees neighboured the same or different species to study mixed species effects. We found that ECM fungal community composition changed and the numbers of vital mycorrhizae decreased for both tree species over 3 drought years (2014-2016), with the ECM fungal community diversity of beech exhibiting a faster and of spruce a stronger decline. Mixed stands had a positive effect on the ECM fungal community diversity of both tree species after the third drought year. Ectomycorrhizae with long rhizomorphs increased in both species under drought, indicating long-distance water transport. However, there was a progressive decline in the number of vital fine roots during the experiment, resulting in a strong reduction in enzyme activity per unit volume of soil. Hydrolytic enzyme activities of the surviving ectomycorrhizae were stable or stimulated upon drought, but there was a large decline in ECM fungal species with laccase activity, indicating a decreased potential to exploit nutrients bound to phenolic compounds. Thus, the ectomycorrhizae responded to repeated drought by maintaining or increasing their functionality at the individual root level, but were unable to compensate for quantitative losses at the ecosystem level. These findings demonstrate a strong below-ground impact of recurrent drought events in forests.

Mycorrhizal associations and the spatial structure of an old-growth forest community.

Plant-soil feedbacks are known to play a central role in species co-existence, but conceptual frameworks for predicting their magnitude and direction are lacking. We ask whether co-occurring trees that associate with different types of mycorrhizal fungi, which are hypothesized to differ in terms of nutrient use and plant-soil feedbacks, differ in sapling establishment densities and probability of co-occurrence. Given that ectomycorrhizal (ECM) trees typically have fungal structures that protect roots from pathogens whereas arbuscular mycorrhizal (AM) trees do not, we hypothesized that ECM saplings would be clustered around ECM trees, while AM saplings would be suppressed near AM trees. Most previous studies have focused on seedlings, but here we examine whether the spatial signal is evident in later life stages. We measured the spatial associations of ~ 28,000 trees using point pattern analysis in a 25-ha old-growth forest where ECM trees comprised 72% of total basal area and 42% of the total stems, while AM trees comprised the remainder. Supporting our hypothesis, AM saplings were more inhibited by AM trees, while ECM saplings were more clustered around ECM trees. The spatial patterns of AM and ECM trees on saplings of the alternate mycorrhizal type were inhibited. To the extent that similar types of feedbacks occur for other AM and ECM trees, our results suggest that fundamental differences in the nature of local-scale biotic interactions between trees and their fungal symbionts may influence forest community assembly and ecosystem dynamics.

Fagus sylvatica seedlings show provenance differentiation rather than adaptation to soil in a transplant experiment.

BACKGROUND: Understanding and predicting the response of tree populations to climate change requires understanding the pattern and scale of their adaptation. Climate is often considered the major driver of local adaptation but, although biotic factors such as soil pathogens or mutualists could be as important, their role has typically been neglected. Biotic drivers might also interact with climate to affect performance and mycorrhizae, in particular, are likely to play a key role in determining drought resistance, which is important in the context of adaptation to future environmental change. To address these questions, we performed a fully reciprocal soil-plant transplant experiment using Fagus sylvatica seedlings and soils from three regions in Germany. To separate the biotic and abiotic effects of inoculation, half of the plants were inoculated with natural soil from the different origins, while the rest were grown on sterilized substrate. We also imposed a drought stress treatment to test for interactions between soil biota and climate. After 1 year of growth, we measured aboveground biomass of all seedlings, and quantified mycorrhizal colonization for a subset of the seedlings, which included all soil-plant combinations, to disentangle the effect of mycorrhiza from other agents. RESULTS: We found that plant origin had the strongest effect on plant performance, but this interacted with soil origin. In general, trees showed a slight tendency to produce less aboveground biomass on local soils, suggesting soil antagonists could be causing trees to be maladapted to their local soils. Consistently, we found lower mycorrhizal colonization rate under local soil conditions. Across all soils, seedlings from low elevations produced more annual biomass than middle (+ 290%) and high (+ 97%) elevations. Interestingly, mycorrhizal colonization increased with drought in the two provenances that showed higher drought tolerance, which supports previous results showing that mycorrhizae can increase drought resistance. CONCLUSIONS: Our findings suggest that soil communities play a role in affecting early performance of temperate trees, although this role may be smaller than that of seed origin. Also, other effects, such as the positive response to generalists or negative interactions with soil biota may be as important as the highly specialized mycorrhizal associations.

The potential for mycobiont sharing between shrubs and seedlings to facilitate tree establishment after wildfire at Alaska arctic treeline.

Root-associated fungi, particularly ectomycorrhizal fungi (EMF), are critical symbionts of all boreal tree species. Although climatically driven increases in wildfire frequency and extent have been hypothesized to increase vegetation transitions from tundra to boreal forest, fire reduces mycorrhizal inoculum. Therefore, changes in mycobiont inoculum may potentially limit tree-seedling establishment beyond current treeline. We investigated whether ectomycorrhizal shrubs that resprout after fire support similar fungal taxa to those that associate with tree seedlings that establish naturally after fire. We then assessed whether mycobiont identity correlates with the biomass or nutrient status of these tree seedlings. The majority of fungal taxa observed on shrub and seedling root systems were EMF, with some dark septate endophytes and ericoid mycorrhizal taxa. Seedlings and adjacent shrubs associated with similar arrays of fungal taxa, and there were strong correlations between the structure of seedling and shrub fungal communities. These results show that resprouting postfire shrubs support fungal taxa compatible with tree seedlings that establish after wildfire. Shrub taxon, distance to the nearest shrub and fire severity influenced the similarity between seedling and shrub fungal communities. Fungal composition was correlated with both foliar C:N ratio and seedling biomass and was one of the strongest explanatory variables predicting seedling biomass. While correlative, these results suggest that mycobionts are important to nutrient acquisition and biomass accrual of naturally establishing tree seedlings at treeline and that mycobiont taxa shared by resprouting postfire vegetation may be a significant source of inoculum for tree-seedling establishment beyond current treeline.

Multiyear fate of a 15 N tracer in a mixed deciduous forest: retention, redistribution, and differences by mycorrhizal association.

The impact of atmospheric nitrogen deposition on forest ecosystems depends in large part on its fate. Past tracer studies show that litter and soils dominate the short-term fate of added 15 N, yet few have examined its longer term dynamics or differences among forest types. This study examined the fate of a 15 N-NO3- tracer over 5-6 years in a mixed deciduous stand that was evenly composed of trees with ectomycorrhizal and arbuscular mycorrhizal associations. The tracer was expected to slowly mineralize from its main initial fate in litter and surface soil, with some 15 N moving to trees, some to deeper soil, and some net losses. Recovery of added 15 N in trees and litterfall totaled 11.3% both 1 and 5-6 years after the tracer addition, as 15 N redistributed from fine and especially coarse roots into cumulative litterfall and small accumulations in woody tissues. Estimates of potential carbon sequestration from tree 15 N recovery amounted to 12-14 kg C per kg of N deposition. Tree 15 N acquisition occurred within the first year after the tracer addition, with no subsequent additional net transfer of 15 N from detrital to plant pools. In both years, ectomycorrhizal trees gained 50% more of the tracer than did trees with arbuscular mycorrhizae. Much of the 15 N recovered in wood occurred in tree rings formed prior to the 15 N addition, demonstrating the mobility of N in wood. Tracer recovery rapidly decreased over time in surface litter material and accumulated in both shallow and deep soil, perhaps through mixing by earthworms. Overall, results showed redistribution of tracer 15 N through trees and surface soils without any losses, as whole-ecosystem recovery remained constant between 1 and 5-6 years at 70% of the 15 N addition. These results demonstrate the persistent ecosystem retention of N deposition even as it redistributes, without additional plant uptake over this timescale.

Oak protein profile alterations upon root colonization by an ectomycorrhizal fungus.

An increased knowledge on the real impacts of ectomycorrhizal symbiosis in forest species is needed to optimize forest sustainable productivity and thus to improve forest services and their capacity to act as carbon sinks. In this study, we investigated the response of an oak species to ectomycorrhizae formation using a proteomics approach complemented by biochemical analysis of carbohydrate levels. Comparative proteome analysis between mycorrhizal and nonmycorrhizal cork oak plants revealed no differences at the foliar level. However, the protein profile of 34 unique oak proteins was altered in the roots. Consistent with the results of the biochemical analysis, the proteome analysis of the mycorrhizal roots suggests a decreasing utilization of sucrose for the metabolic activity of mycorrhizal roots which is consistent with an increased allocation of carbohydrates from the plant to the fungus in order to sustain the symbiosis. In addition, a promotion of protein unfolding mechanisms, attenuation of defense reactions, increased nutrient mobilization from the plant-fungus interface (N and P), as well as cytoskeleton rearrangements and induction of plant cell wall loosening for fungal root accommodation in colonized roots are also suggested by the results. The suggested improvement in root capacity to take up nutrients accompanied by an increase of root biomass without apparent changes in aboveground biomass strongly re-enforces the potential of mycorrhizal inoculation to improve cork oak forest resistance capacity to cope with coming climate change.

Multiple Friends with Benefits: An Optimal Mutualist Management Strategy?

Most mutualisms in nature involve interactions between multispecies mutualist guilds and multiple partner species. While mechanisms such as niche partitioning can explain part of this diversity, the presence of low-quality partners, which produce relatively low returns on investment compared with other guild members, is not well understood. Here, we consider a novel explanation for this persistence: that low-quality partners are actively maintained by their hosts as part of a growth-maximizing strategy, even in the presence of higher-quality alternatives. We use a model inspired by the interaction between host trees and ectomycorrhizal fungi to demonstrate that when the environment is variable, trees maintain low-quality fungal partners that they would not otherwise maintain in constant environments. This active investment, which emerges as a response to saturating returns on investment in higher-quality partners, could contribute to the maintenance of diversity in multispecies mutualisms.