Symbiotic Ectomycorrhizal Fungi Research Articles

The ectomycorrhizal fungus Scleroderma bovista improves growth of hazelnut seedlings and plays a role in auxin signaling and transport.

Scleroderma bovista can form symbiotic ectomycorrhizal fungi with hazel roots. The mechanism through which S. bovista promotes hazelnut growth remains unclear. This study aimed to evaluate the effect of ectomycorrhizal fungus S. bovista on the growth and development of hazel roots and gene expression changes through comparative transcriptome analysis. After inoculation with S. bovista, the fungus symbiotically formed ectomycorrhiza with hazel roots. The fresh weights of the aboveground and underground parts of My treatment (inoculated with S. bovista and formed mycorrhiza) were much higher than those of the control, respectively. The length, project area, surface area, volume, forks, and diameter of the inoculated seedlings root were 1.13 to 2.48 times higher than those of the control. In the paired comparison, 3,265 upregulated and 1,916 downregulated genes were identified. The most significantly enriched Gene Ontology term for the upregulated Differentially Expressed Genes was GO:0005215 (transporter activity). Immunohistochemical analysis suggested that the expression levels of auxin and Auxin Response Factor9 were significantly increased by S. bovista after the formation of mycorrhizal fungi in hazelnut root tips. These results indicate that genes related to auxin biosynthesis, transport and signaling, and transport of nutrients may contribute to root development regulation in hazel ectomycorrhiza.

Relationships between rhizosphere microbial communities, soil abiotic properties and root trait variation within a pine species

Abstract Rhizosphere microbes play important roles in plant performance and ecosystem functioning. It is becoming increasingly clear that rhizosphere communities vary with soil properties and variation in root traits among plant species. However, less is known about whether and how variation in root traits within plant species influences the rhizosphere microbial communities. We evaluated the intraspecific root traits variation and explored their associations with bacterial and fungal communities in rhizosphere by focusing on an ectomycorrhizal tree species, that is Pinus massoniana, in 22 sites in subtropical China. The first dimension of the principal component analysis on root traits revealed evidence for the ‘conservation’ gradient of the root economics space. Overall, root traits explained more variation in fungal communities than in bacterial communities in the rhizosphere. Functional composition of rhizosphere microbial communities changed significantly along the ‘conservation’ gradient, with fast‐growing copiotrophic bacteria and symbiotic ectomycorrhizal fungi were significantly enriched on the ‘acquisition’ side, while slow‐growing oligotrophic bacteria were significantly enriched on the ‘conservation’ side of the gradient. Synthesis: Our study demonstrates that intraspecific variation in plant roots significantly influence rhizosphere microbial communities, which in turn can influence plant nutrition and therefore plant performance within the community.

Benefits of symbiotic ectomycorrhizal fungi to plant water relations depend on plant genotype in pinyon pine

Rhizosphere microbes, such as root-associated fungi, can improve plant access to soil resources, affecting plant health, productivity, and stress tolerance. While mycorrhizal associations are ubiquitous, plant–microbe interactions can be species specific. Here we show that the specificity of the effects of microbial symbionts on plant function can go beyond species level: colonization of roots by ectomycorrhizal fungi (EMF) of the genus Geopora has opposite effects on water uptake, and stomatal control of desiccation in drought tolerant and intolerant genotypes of pinyon pine (Pinus edulis Engelm.). These results demonstrate, for the first time, that microorganisms can have significant and opposite effects on important plant functional traits like stomatal control of desiccation that are associated with differential mortality and growth in nature. They also highlight that appropriate pairing of plant genotypes and microbial associates will be important for mitigating climate change impacts on vegetation.

Woodland wildfire enables fungal colonization of encroaching Douglas‐fir

Abstract Self‐reinforcing differences in fire frequency help closed‐canopy forests, which resist fire, and open woodlands, which naturally burn often, to co‐occur stably at landscape scales. Forest tree seedlings, which could otherwise encroach and overgrow woodlands, are killed by regular fire, yet fire has other effects that may also influence these feedbacks. In particular, many forest trees require symbiotic ectomycorrhizal fungi in order to establish. By restructuring soil fungal communities, fire might affect the availability of symbionts or the potential for symbiont sharing between encroaching trees and woodland vegetation. To investigate this possibility, we performed a soil bioassay experiment using inoculum from burned and unburned oak woodlands and Douglas‐fir forests. We examined how fire, ecosystem type, and neighboring heterospecific seedlings affect fungal root community assembly of Douglas‐firs and oaks. We asked whether heterospecific seedlings facilitated fungal colonization of seedling roots in non‐native soil, and if so, whether fire influenced this interaction. External fungal colonization of oak roots was more influenced by fire and ecosystem type than by the presence of a Douglas‐fir, and oaks increased the likelihood that Douglas‐fir roots would be colonized by fungi in oak woodland soil. Yet, fire increased colonization of Douglas‐fir in oak soil, diminishing the otherwise crucial role played by oak facilitation. Fire also strengthened the positive effect of Douglas‐firs on oak root‐associated fungal diversity in Douglas‐fir forest soil. Prior work shows that fire supports woodland ecosystems by stemming recruitment of encroaching seedlings. Here, we find evidence that it may contrastingly reduce fungal limitation of invasive seedling growth and establishment, otherwise relieved only by facilitation. Future work can investigate how these opposing effects might contribute to the net impact of changes in fire regime on landcover stability. Read the free Plain Language Summary for this article on the Journal blog.

Cloning of Nitrate Reductase and Nitrite Reductase Genes and Their Functional Analysis in Regulating Cr(VI) Reduction in Ectomycorrhizal Fungus Pisolithus sp.1.

Assimilatory-type nitrate reductase (NR) and nitrite reductase (NiR) are the key enzymes that involve in nitrate assimilation and nitrogen cycling in microorganisms. NR and NiR with NADH or NADPH and FMN or FAD domains could be coupled to the reduction process of hexavalent chromium [Cr(VI)] in microorganisms. A new assimilatory-type NR gene (named niaD) and a new assimilatory-type NiR gene (named niiA) are cloned, identified, and functionally characterized by 5′ and 3′ RACE, alignment, annotation, phylogenetic tree, and yeast mutant complementation analyses from Pisolithus sp.1, a dominant symbiotic ectomycorrhizal fungi (EMF) that can assist in phytoremediation. Assimilatory-type niaD and niiA were 2,754 bp and 3,468 bp and encode a polypeptide with 917 and 1,155 amino acid residues, respectively. The isoelectric points of NR (Pisolithus sp.1 NR) and NiR (Pisolithus sp.1 NiR) of Pisolithus sp.1 are 6.07 and 6.38, respectively. The calculated molecular mass of Pisolithus sp.1 NR and Pisolithus sp.1 NiR is 102.065 and 126.914 kDa, respectively. Yeast mutant complementation analysis, protein purification, and activities of NR and NiR under Cr treatment suggest that Pisolithus sp.1 NR is a functional NR that mediates Cr(VI) tolerance and reduction. The multiple alignment demonstrates that Pisolithus sp.1 NR is potentially a nicotinamide adenine dinucleotide phosphate-dependent flavin mononucleotide reductase and also Class II chromate reductase. Our results suggest that Pisolithus sp.1 NR plays a key role in Cr(VI) reduction in the EMF Pisolithus sp.1.

Ectomycorrhiza-hydrogel additive enhanced growth of Norway spruce seedlings in a nutrient-poor peat substrate

Seedling quality is an important input affecting the outplanted seedling performance. Morphological attributes and association with symbiotic ectomycorrhizal (ECM) fungi belong to influential traits determining seedling quality. In this study, the effect of pre-sowing applied commercial additives Ectovit (containing hydrogel and ECM fungi), Mycorrhizaroots (ECM fungi and nutrients) and Vetozen (natural mineral fertilizer) on the growth and ectomycorrhiza formation of Norway spruce seedlings grown in nutrient-poor pure peat in an open nursery site was assessed. Two-year-old bareroot seedlings were transplanted into containers. No significant growth differences were detected between treatments for 1 + 0 seedlings but the values of growth parameters (root collar diameter; stem height; shoot, root and total dry weight) of Ectovit-inoculated seedlings were significantly higher than those in the other treatments including the control after the second (2 + 0 seedlings) and the third growing season (2 + 1). Root-to-shoot dry weight ratio and number of root tips were distinctly higher after the third year compared to the previous two years but they were not significantly different between treatments. Mycorrhizaroots and Vetozen did not have any significant effect on seedling development. Although the occurrence of treatment-specific ECM morphotype was detected in Ectovit-inoculated seedlings, the high total ECM colonization of roots in all treatments including the control indicated a crucial impact of naturally occurring fungi on ectomycorrhiza formation.

Direct and indirect effects of fire on microbial communities in a pyrodiverse dry‐sclerophyll forest

Abstract Fire is one of the predominant drivers of the structural and functional dynamics of forest ecosystems. In recent years, novel fire regimes have posed a major challenge to the management of pyrodiverse forests. While previous research efforts have focused on quantifying the impacts of fire on above‐ground forest biodiversity, how microbial communities respond to fire is less understood, despite their functional significance. Here, we describe the effects of time since fire, fire frequency and their interaction on soil and leaf litter fungal and bacterial communities from the pyrodiverse, Eucalyptus pilularis forests of south‐eastern Australia. Using structural equation models, we also elucidate how fire can influence these communities both directly and indirectly through biotic–abiotic interactions. Our results demonstrate that fire is a key driver of litter and soil bacterial and fungal communities, with effects most pronounced for soil fungal communities. Notably, recently burnt forest hosted lower abundances of symbiotic ectomycorrhizal fungi and Acidobacteria in the soil, and basidiomycetous fungi and Actinobacteriota in the litter. Compared with low fire frequencies, high fire frequency increased soil fungal plant pathogens, but reduced Actinobacteriota. The majority of fire effects on microbial communities were mediated by fire‐induced changes in litter and soil abiotic properties. For instance, recent and more frequent fire was associated with reduced soil sulphur, which led to an increase in soil fungal plant pathogens and saprotrophic fungi in these sites. Pathogenic fungi also increased in recently burnt forests that had a low fire frequency, mediated by a decline in litter carbon and an increase in soil pH in these sites. Synthesis. Our findings indicate that predicted increases in the frequency of fire may select for specific microbial communities directly and indirectly through ecological interactions, which may have functional implications for plants (increase in pathogens, decrease in symbionts), decomposition rates (declines in Actinobacteriota and Acidobacteriota) and carbon storage (decrease in ectomycorrhizal fungi). In the face of predicted shifts in wildfire regimes, which may exacerbate fire‐induced changes in microbial communities, adaptive fire management and monitoring is required to address the potential functional implications of fire‐altered microbial communities.

Predictors of taxonomic and functional composition of black spruce seedling ectomycorrhizal fungal communities along peatland drainage gradients.

Many trees depend on symbiotic ectomycorrhizal fungi for nutrients in exchange for photosynthetically derived carbohydrates. Trees growing in peatlands, which cover 3% of the earth's terrestrial surface area yet hold approximately one-third of organic soil carbon stocks, may benefit from ectomycorrhizal fungi that can efficiently forage for nutrients and degrade organic matter using oxidative enzymes such as class II peroxidases. However, such traits may place a higher carbon cost on both the fungi and host tree. To investigate these trade-offs that might structure peatland ectomycorrhizal fungal communities, we sampled black spruce (Picea mariana (Mill.)) seedlings along 100-year-old peatland drainage gradients in Minnesota, USA, that had resulted in higher soil nitrogen and canopy density. Structural equation models revealed that the relative abundance of the dominant ectomycorrhizal fungal genus, Cortinarius, which is known for relatively high fungal biomass coupled with elevated class II peroxidase potential, was negatively linked to site fertility but more positively affected by recent host stem radial growth, suggesting carbon limitation. In contrast, Cenococcum, known for comparatively lower fungal biomass and less class II peroxidase potential, was negatively linked to host stem radial growth and unrelated to site fertility. Like Cortinarius, the estimated relative abundance of class II peroxidase genes in the ectomycorrhizal community was more related to host stem radial growth than site fertility. Our findings indicate a trade-off between symbiont foraging traits and associated carbon costs that consequently structure seedling ectomycorrhizal fungal communities in peatlands.

Spheres of Influence: Host Tree Proximity and Soil Chemistry Shape rRNA, but Not DNA, Communities of Symbiotic and Free-Living Soil Fungi in a Mixed Hardwood-Conifer Forest

Host and symbiont diversity are inextricably linked across partnerships and ecosystems, with degree of partner reliance governing the strength of this correlation. In many forest soils, symbiotic ectomycorrhizal fungi coexist and compete with free-living saprotrophic fungi, with the outcomes of these interactions shaping resource availability and competitive outcomes for the trees aboveground. Traditional approaches to characterizing these communities rely on DNA sequencing of a ribosomal precursor RNA gene (the internal transcribed spacer region), but directly sequencing the precursor rRNA may provide a more functionally relevant perspective on the potentially active fungal communities. Here, we map ectomycorrhizal and saprotrophic soil fungal communities through a mixed hardwood-conifer forest to assess how above- and belowground diversity linkages compare across these differently adapted guilds. Using highly spatially resolved transects (sampled every 2 m) and well-mapped stands of varying host tree diversity, we sought to understand the relative influence of symbiosis versus environment in predicting fungal diversity measures. Canopy species in this forest included two oaks (Quercus agrifoliaandQuercus douglasii) and one pine (Pinus sabiniana). At the scale of our study, spatial turnover in rRNA-based communities was much more predictable from measurable environmental attributes than DNA-based communities. And while turnover of ectomycorrhizal fungi and saprotrophs were predictable by the presence and abundance of different canopy species, they both responded strongly to soil nutrient characteristics, namely pH and nitrogen availability, highlighting the niche overlap of these coexisting guilds and the strong influence of aboveground plants on belowground fungal communities.

Shifts in ectomycorrhizal exploration types parallel leaf and fine root area with forest age

Abstract The abundance of fine roots and leaves in forests typically peaks during mid‐succession and then declines. If fine root area declines more rapidly than leaf area, this could contribute disproportionately to stand decline. However, trees also partner with symbiotic ectomycorrhizal (EcM) fungi that facilitate nutrient acquisition of fine roots. Ectomycorrhizal fungi can use the carbohydrates provided by their plant partners to produce emanating fungal tissues which increase soil exploration potential but may also prove costly. We suggest two competing hypotheses to frame the response of ectomycorrhizas to stand age: (a) the ‘functional offset’ hypothesis posits that the abundance of ectomycorrhizas with emanating tissues (‘Distance’ mycorrhizas) increases with stand age and (b) the ‘energy‐limited’ hypothesis posits that carbon available for root symbionts decreases with stand age resulting in fewer Distance mycorrhizas. In the first hypothesis, EcM functional traits offset root abundance, while in the second, traits parallel root abundance. To test these competing hypotheses, we sampled fine roots to a depth of 90 cm and used allometric equations to estimate changes in root and leaf area index across a chronosequence of Pinus banksiana stands ranging from 2 to 76 years average tree age. We also examined fine roots microscopically to track changes in the abundance of EcM functional types. We used DNA‐based methods to sequence EcM fungi and confirm roots were of P. banksiana. Both fine root and leaf area of pine increased for the first 30–36 years (until Mid age) and then plateaued, while the ratio of leaf to fine root area was similar across the age gradient. Also, changes to fine root area with stand age depended on soil depth. The abundance of Distance mycorrhizas was lowest at the youngest plots and increased until Mid age, where it peaked, contrary to the functional offset hypothesis. Instead, the abundance of Distance mycorrhizas paralleled changes to leaf area, aligned with the energy‐limited hypothesis. Synthesis. Ectomycorrhizas do not offset the function of roots, rather shifts in exploration type likely reflect adjustments to carbon supply from hosts.

Shifts in ectomycorrhizal fungal community composition during the early establishment of native and exotic pine seedlings

Exotic pine trees are often introduced to novel places worldwide. How exotic pines interact with local microbial communities, such as symbiotic ectomycorrhizal (ECM) fungi, during early establishment compared to interactions by their native counterparts remains to be examined in detail. In this study, native masson pine (Pinus massoniana) and exotic slash pine (Pinus elliottii) seedlings were grown in field-collected soil cores. ECM root tips were collected 3- and 6-months after planting and fungi were identified using high-throughput sequencing. We found that host identity was a key factor determining ECM fungal community assembly after 3-months: native pines associated with Rhizopogon while exotic pines associated primarily with ECM generalists. After 6-months growth, however, ECM fungal communities in native and exotic pines were both dominated by Rhizopogon species. We also found that host selections of several pioneer ECM fungi shared by both native and exotic pines may facilitate an exotic pine's establishment. These findings indicate that more attention should be paid to ECM fungal community shuffling by exotic trees during early establishment and, in turn, the influence of the potential changes in ECM fungal communities on local forest ecosystems.

Plant intraspecific variation modulates nutrient cycling through its below ground rhizospheric microbiome

Abstract Plant genetic variation, through its phenotypic display, can determine the composition of below ground microbial communities. Variation within a species is increasingly acknowledged to have substantial ecological consequences, particularly through trophic cascades. We hypothesized that the intraspecific genotypic variation of the tree host might impact the phylogenetic composition of its rhizospheric microbial communities, by favouring particular clades, that might be further reflected in ecosystem process rates. We tested whether the intraspecific genotypic variation of Pinus pinaster modulates nutrient cycling by determining the phylogenetic structure of its symbiotic ectomycorrhizal fungi and rhizospheric bacteria. We sequenced fungal and bacterial molecular markers and reconstructed phylogenies in the rhizosphere of P. pinaster trees belonging to three genotypic variants (Mediterranean, Atlantic, African) in three 45‐year‐old common garden experiments, and measured seven soil enzymatic activities. Local effects, based on differences in elevation and soil conditions across sites, were strong predictors of the ectomycorrhizal and bacterial communities thriving in tree’s rhizosphere. Across‐site variation also explained differences in phosphorus cycling. We detected, however, a significant effect of the plant genotype on the phylogenetic structure of the root‐associated microbiota that was consistent across sites. The most productive Mediterranean plant genotype sheltered the most distinct root microbiome, with the dominant Basidiomycetes and Proteobacteria having a strong influence on the phylogenetic microbial community structure and associating with an enhanced hydrolysis of celluloses, hemicelluloses and chitin. Beneath the less productive Atlantic genotype, the less abundant Ascomycetes and up to thirteen bacterial phyla shaped the phylogenetic microbial structure, and predicted the rates of peptidase. Ectomycorrhizal fungi explained the activity of cellulases and protease, and bacteria that of hemicellulases and chitinase, suggesting functional complementarity. Synthesis. This is the first report using three‐replicated long‐term common gardens in mature forests to disentangle plant genotype‐ and site‐specific drivers of the rhizospheric microbiome and its enzymatic potential. We concluded that intraspecific variation in primary producers leaves a phylogenetic signature in mutualists and decomposers that further modulate key steps in carbon and nitrogen cycles. These results emphasize the ecological relevance of plant intraspecific diversity in determining essential plant–soil feedbacks that control ecosystem productivity and performance.

Correction: Metatranscriptomic Study of Common and Host-Specific Patterns of Gene Expression between Pines and Their Symbiotic Ectomycorrhizal Fungi in the Genus Suillus.

[This corrects the article DOI: 10.1371/journal.pgen.1006348.].

Genetically determined fungal pathogen tolerance and soil variation influence ectomycorrhizal traits of loblolly pine.

Selection on genetically correlated traits within species can create indirect effects on one trait by selection on another. The consequences of these trait correlations are of interest because they may influence how suites of traits within species evolve under differing selection pressures, both natural and artificial. By utilizing genetic families of loblolly pine either tolerant (t) or susceptible (s) to two different suites of pathogenic fungi responsible for causing either pine decline or fusiform rust disease, we investigated trait variation and trait correlations within loblolly pine (Pinus taeda L.) by determining how ectomycorrhizal (EM) colonization relates to pathogen susceptibility. We detected interactions between susceptibility to pathogenic fungi and soil inoculation source on loblolly pine compatibility with the EM fungi Thelephora, and on relative growth rate of loblolly pine. Additionally, we detected spatial variation in the loblolly pine–EM fungi interaction, and found that variation in colonization rates by some members of the EM community is not dictated by genetic variation in the host plant but rather soil inoculation source alone. The work presented here illustrates the potential for indirect selection on compatibility with symbiotic EM fungi as a result of selection for resistance to fungal pathogens. Additionally, we present evidence that the host plant does not have a single “mycorrhizal trait” governing interactions with all EM fungi, but rather that it can interact with different fungal taxa independently. Synthesis. An understanding of the genetic architecture of essential traits in focal species is crucial if we are to anticipate and manage the results of natural and artificial selection. As demonstrated here, an essential but often overlooked symbiosis (that between plants and mycorrhizal fungi) may be indirectly influenced by directed selection on the host plant.

Saprotrophic and ectomycorrhizal fungal sporocarp stoichiometry (C : N : P) across temperate rainforests as evidence of shared nutrient constraints among symbionts.

Quantifying nutritional dynamics of free-living saprotrophs and symbiotic ectomycorrhizal fungi in the field is challenging, but the stoichiometry of fruiting bodies (sporocarps) may be an effective methodology for this purpose. Carbon (C), nitrogen (N) and phosphorus (P) concentrations of soils, foliage and 146 sporocarp collections were analyzed from 14 Pseudotsuga menziesii var. menziesii stands across a podzolization gradient on Vancouver Island (Canada). N and P concentrations were considerably higher in saprotrophic fungi. Fungal N% increased with soil N content at a greater rate for saprotrophs than ectomycorrhizal fungi, while fungal P% of saprotrophs was more constrained. Fungal N : P was more responsive to soil N : P for ectomycorrhizal fungi (homeostatic regulation coefficient 'H' = 2.9) than saprotrophs (H = 5.9), while N : P of ectomycorrhizal fungi and host tree foliage scaled almost identically. Results underscore the role of ectomycorrhizal fungi as nutrient conduits, supporting host trees, whereas saprotrophs maintain a greater degree of nutritional homeostasis. Site nutrient constraints were shared in equal measure between ectomycorrhizal fungi and host trees, particularly for P, suggesting neither partner benefits from enhanced nutrition at the expense of the other. Sporocarp stoichiometry provides new insights into mycorrhizal relationships and illustrates pervasive P deficiencies across temperate rainforests of the Pacific Northwest.

Contrasting effects of ectomycorrhizal fungi on early and late stage decomposition in a boreal forest.

Symbiotic ectomycorrhizal fungi have received increasing attention as regulators of below-ground organic matter storage. They are proposed to promote organic matter accumulation by suppressing saprotrophs, but have also been suggested to play an active role in decomposition themselves. Here we show that exclusion of tree roots and associated ectomycorrhizal fungi in a boreal forest increased decomposition of surface litter by 11% by alleviating nitrogen limitation of saprotrophs-a "Gadgil effect". At the same time, root exclusion decreased Mn-peroxidase activity in the deeper mor layer by 91%. Our results show that ectomycorrhizal fungi may hamper short-term litter decomposition, but also support a crucial role of ectomycorrhizal fungi in driving long-term organic matter oxidation. These observations stress the importance of ectomycorrhizal fungi in regulation of below-ground organic matter accumulation. By different mechanisms they may either hamper or stimulate decomposition, depending upon stage of decomposition and location in the soil profile.

Early-successional ectomycorrhizal fungi effectively support extracellular enzyme activities and seedling nitrogen accumulation in mature forests.

After stand-replacing disturbance, regenerating conifer seedlings become colonized by different ectomycorrhizal fungi (EMF) than the locally adapted EMF communities present on seedlings in mature forests. We studied whether EMF species that colonized subalpine fir (Abies lasiocarpa) seedlings in clearcuts differed from those that colonized seedlings in adjacent mature forests with respect to mycorrhizoplane extracellular enzyme activities (EEAs) and N status of the seedlings. We tested two alternate hypotheses: (1) that EEAs would differ between the two EMF communities, with higher activities associated with forest-origin communities, and (2) that acclimation to soil environment was considerable enough that EEAs would be determined primarily by the soil type in which the ectomycorrhizas were growing. Naturally colonized fir seedlings were reciprocally transplanted between clearcuts and forests, carrying different EMF communities with them. EEAs were influenced more by destination environment than by EMF community. EEAs were as high in early-successional as in late-successional communities in both destination environments. Buds of clearcut-origin seedlings had the same or higher N contents as forest seedlings after a growing season in either environment. These results indicate that (i) symbiotic EMF and/or their associated microbial communities demonstrate substantial ability to acclimate to new field environments; (ii) the ability to produce organic matter-degrading enzymes is not a trait that necessarily distinguishes early- and late-successional EMF communities in symbiosis; (iii) early-successional EMF are as capable of supporting seedling N accumulation in forest soils as late-successional EMF; and (iv) disturbed ecosystems where early-successional EMF are present should have high resilience for organic matter degradation.

Metatranscriptomic Study of Common and Host-Specific Patterns of Gene Expression between Pines and Their Symbiotic Ectomycorrhizal Fungi in the Genus Suillus.

Ectomycorrhizal fungi (EMF) represent one of the major guilds of symbiotic fungi associated with roots of forest trees, where they function to improve plant nutrition and fitness in exchange for plant carbon. Many groups of EMF exhibit preference or specificity for different plant host genera; a good example is the genus Suillus, which grows in association with the conifer family Pinaceae. We investigated genetics of EMF host-specificity by cross-inoculating basidiospores of five species of Suillus onto ten species of Pinus, and screened them for their ability to form ectomycorrhizae. Several Suillus spp. including S. granulatus, S. spraguei, and S. americanus readily formed ectomycorrhizae (compatible reaction) with white pine hosts (subgenus Strobus), but were incompatible with other pine hosts (subgenus Pinus). Metatranscriptomic analysis of inoculated roots reveals that plant and fungus each express unique gene sets during incompatible vs. compatible pairings. The Suillus-Pinus metatranscriptomes utilize highly conserved gene regulatory pathways, including fungal G-protein signaling, secretory pathways, leucine-rich repeat and pathogen resistance proteins that are similar to those associated with host-pathogen interactions in other plant-fungal systems. Metatranscriptomic study of the combined Suillus-Pinus transcriptome has provided new insight into mechanisms of adaptation and coevolution of forest trees with their microbial community, and revealed that genetic regulation of ectomycorrhizal symbiosis utilizes universal gene regulatory pathways used by other types of fungal-plant interactions including pathogenic fungal-host interactions.

Fractionation and assimilation of Mg isotopes by fungi is species dependent.

Symbiotic ectomycorrhizal fungi mobilize nutrients from both organic and inorganic substrates and supply them to their host plants. Their role in mobilizing base cations and phosphorus from mineral substrates through weathering has received increasing attention in recent years but the processes involved remain to be elucidated. We grew selected ectomycorrhizal and nonmycorrhizal fungi in axenic systems containing mineral and organic substrates and examined their capacity to fractionate and assimilate stable isotopes of magnesium. The mycorrhizal fungi were significantly depleted in heavy isotopes with the lowest Δ26 Mg values (the difference between δ26 Mg in fungal tissue and δ26 Mg in the substrate) compared with nonmycorrhizal fungi, when grown on mineral substrates containing granite particles. The ectomycorrhizal fungi accumulated significantly higher concentrations of Mg, K and P than the nonmycorrhizal fungi. There was a highly significant statistical relationship between δ26 Mg tissue signature and mycelial concentration of Mg, with a clear separation between most ectomycorrhizal fungi and the nonmycorrhizal fungi. These results are consistent with the idea that ectomycorrhizal fungi have evolved efficient mechanisms to mobilize, transport and store Mg within their mycelia.

Ectomycorrhizal fungi slow soil carbon cycling.

Respiration of soil organic carbon is one of the largest fluxes of CO2 on earth. Understanding the processes that regulate soil respiration is critical for predicting future climate. Recent work has suggested that soil carbon respiration may be reduced by competition for nitrogen between symbiotic ectomycorrhizal fungi that associate with plant roots and free-living microbial decomposers, which is consistent with increased soil carbon storage in ectomycorrhizal ecosystems globally. However, experimental tests of the mycorrhizal competition hypothesis are lacking. Here we show that ectomycorrhizal roots and hyphae decrease soil carbon respiration rates by up to 67% under field conditions in two separate field exclusion experiments, and this likely occurs via competition for soil nitrogen, an effect larger than 2°C soil warming. These findings support mycorrhizal competition for nitrogen as an independent driver of soil carbon balance and demonstrate the need to understand microbial community interactions to predict ecosystem feedbacks to global climate.