Ectomycorrhizal Fungus Paxillus Involutus Research Articles

Elucidating fungal decomposition of organic matter at sub-micrometer spatial scales using optical photothermal infrared (O-PTIR) microspectroscopy.

In microbiological studies, a common goal is to link environmental factors to microbial activities. Both environmental factors and microbial activities are typically derived from bulk samples. It is becoming increasingly clear that such bulk environmental parameters poorly represent the microscale environments microorganisms experience. Using infrared (IR) microspectroscopy, the spatial distribution of chemical compound classes can be visualized, making it a useful tool for studying the interactions between microbial cells and their microenvironments. The spatial resolution of conventional IR microspectroscopy has been limited by the diffraction limit of IR light. The recent development of optical photothermal infrared (O-PTIR) microspectroscopy has pushed the spatial resolution of IR microspectroscopy beyond this diffraction limit, allowing the distribution of chemical compound classes to be visualized at sub-micrometer spatial scales. To examine the potential and limitations of O-PTIR microspectroscopy to probe the interactions between fungal cells and their immediate environments, we imaged the decomposition of cellulose films by cells of the ectomycorrhizal fungus Paxillus involutus and compared O-PTIR results using conventional IR microspectroscopy. Whereas the data collected with conventional IR microspectroscopy indicated that P. involutus has only a very limited ability to decompose cellulose films, O-PTIR data suggested that the ability of P. involutus to decompose cellulose was substantial. Moreover, the O-PTIR method enabled the identification of a zone located outside the fungal hyphae where the cellulose was decomposed by oxidation. We conclude that O-PTIR can provide valuable new insights into the abilities and mechanisms by which microorganisms interact with their surrounding environments.IMPORTANCEInfrared (IR) microspectroscopy allows the spatial distribution of chemical compound classes to be visualized. The use of conventional IR microspectroscopy in microbiological studies has been restricted by limited spatial resolution. Recent developments in laser technology have enabled a new class of IR microspectroscopy instruments to be developed, pushing the spatial resolution beyond the diffraction limit of IR light to approximately 500 nm. This improved spatial resolution now allows microscopic observations of changes in chemical compounds to be made, making IR microspectroscopy a useful tool to investigate microscale changes in chemistry that are caused by microbial activity. We show these new possibilities using optical photothermal infrared microspectroscopy to visualize the changes in cellulose substrates caused by oxidation by the ectomycorrhizal fungus Paxillus involutus at the interface between individual fungal hyphae and cellulose substrates.

Ectomycorrhizal Fungal Transformation of Dissolved Organic Matter: Consequences for Reductive Iron Oxide Dissolution and Fenton-Based Oxidation of Mineral-Associated Organic Matter

Recent studies have shown that dissolved organic matter (DOM) decomposed by ectomycorrhizal (ECM) fungi increases adsorptive properties of organic matter towards soil mineral surfaces. Concomitantly, ECM fungi secrete secondary metabolites with iron reducing capacity that are thought to participate in non-enzymatic Fenton-based decomposition of DOM. The aim of this study was to investigate if the iron reduction induced by the ECM fungus Paxillus involutus during organic matter decomposition was conserved in the decomposed DOM. We explored how the modified DOM reductively dissolved ferrihydrite and goethite nanoparticles and how these processes affected the reactions with H2O2 and the Fenton-based oxidation of mineral-associated organic matter. Culture filtrates were obtained from incubation of the ECM fungus on DOM from forest litter of a spruce forest. This modified DOM was separated by extraction into an ethyl acetate and a water fraction. These fractions were reacted with ferrihydrite and goethite in absence and presence of H2O2. Dissolved Fe2+ and HO• were measured and the reactions at the iron oxide-water interfaces were monitored in real-time with in-situ IR spectroscopy. Experiments showed that decomposition of DOM by P. involutus generated a modified DOM that displayed an increased and persistent reductive capacity. Most of the reductants were isolated in the aromatic- and carboxyl-dominated ethyl acetate fraction but some reduction capacity was also captured in the water fraction mainly containing carbohydrates. Reductive dissolution was more extensive for ferrihydrite than goethite, and this process generated significant oxidation of the DOM-ferrihydrite associations. Oxidation of adsorbed DOM was triggered by H2O2via heterogenous and homogeneous Fenton reactions. These oxidation processes were favored by ferrihydrite because of a high reduction potential and a high efficiency of heterogeneous Fenton as compared to goethite. An optimal timing between the heterogeneous and homogeneous Fenton processes triggered extensive radical oxidation of the DOM-ferrihydrite associations generating a high concentration of surface-associated oxalate. Overall, the results show that organic matter associated with ferrihydrite may be more susceptible to radical oxidation than on goethite, and that fungal decomposition of DOM in general may have consequences for other important soil processes such as mineral dissolution, adsorption and initiation of radical reactions.

Mycorrhizal Mediated Partitioning of Phosphorus: Ectomycorrhizal (Populus x canescens x Paxillus involutus) Potential to Exploit Simultaneously Organic and Mineral Phosphorus Sources

Many natural and anthropogenic soils are phosphorus (P) limited often due to larger P stocks sequestered in forms of low bioavailability. One of the strategies to overcome this shortage lies in the symbiosis of plants with mycorrhizal fungi, increasing the plant P uptake of these hardly accessible sources. However, little is known about mycorrhizal fungal mediated partitioning of differently available P forms, which could contribute to more efficient use of P by plants and, thereby, reduce competition for soil P. This study aimed to investigate the uptake of P from differently bioavailable P sources by ectomycorrhiza. For that, we conducted a rhizotrone study using Populus x canescens and its compatible ectomycorrhizal fungus Paxillus involutus. Four different P sources [ortho-phosphate (oP), adenosine monophosphate (AMP), hydroxyapatite (HAP), and oP bound to goethite (gP)] or only HAP as 1P control were supplied in separate compartments, where only the fungal partner had access to the P sources. The amount of the specific P sources was increased according to their decreasing bioavailability. In order to distinguish between the P sources, we applied 33P to track its incorporation in plants by a non-destructive analysis via digital autoradiography. Our results show that an ectomycorrhizal plant is able to utilize all provided P sources via its mycorrhizal fungal associate. The acquisition timing was determined by the most bioavailable P sources, with oP and AMP over HAP and gP, and a mixed P pool over a single P source. In contrast, the magnitude was defined by the by the amount of supplied P source provision of additional nitrogen, hence AMP over oP and gP, as well as by P source complexity, with gP as the least favorable P form. Nevertheless, the results of the present study provide evidence that an ectomycorrhiza has the potential to occupy fundamental niches of various P sources differing in their bioavailability, indicating that being a generalist in P nutrition can facilitate adaptation to various nutritional settings in soil.

Secretion of Iron(III)-Reducing Metabolites during Protein Acquisition by the Ectomycorrhizal Fungus Paxillus involutus.

The ectomycorrhizal fungus Paxillus involutus decomposes proteins using a two-step mechanism, including oxidation and proteolysis. Oxidation involves the action of extracellular hydroxyl radicals (•OH) generated by the Fenton reaction. This reaction requires the presence of iron(II). Here, we monitored the speciation of extracellular iron and the secretion of iron(III)-reducing metabolites during the decomposition of proteins by P. involutus. X-ray absorption spectroscopy showed that extracellular iron was mainly present as solid iron(III) phosphates and oxides. Within 1 to 2 days, these compounds were reductively dissolved, and iron(II) complexes were formed, which remained in the medium throughout the incubation. HPLC and mass spectrometry detected five extracellular iron(III)-reducing metabolites. Four of them were also secreted when the fungus grew on a medium containing ammonium as the sole nitrogen source. NMR identified the unique iron(III)-reductant as the diarylcyclopentenone involutin. Involutin was produced from day 2, just before the elevated •OH production, preceding the oxidation of BSA. The other, not yet fully characterized iron(III)-reductants likely participate in the rapid reduction and dissolution of solid iron(III) complexes observed on day one. The production of these metabolites is induced by other environmental cues than for involutin, suggesting that they play a role beyond the Fenton chemistry associated with protein oxidation.

Influence of Ammonium on Formation of Mineral-Associated Organic Carbon by an Ectomycorrhizal Fungus.

The interactions between dissolved organic matter (DOM) and mineral particles are critical for the stabilization of soil organic matter (SOM) in terrestrial ecosystems. The processing of DOM by ectomycorrhizal fungi contributes to the formation of mineral-stabilized SOM by two contrasting pathways: the extracellular transformation of DOM (ex vivo pathway) and the secretion of mineral-surface-reactive metabolites (in vivo pathway). In this study, we examined how changes in nitrogen (N) availability affected the formation of mineral-associated carbon (C) from these two pathways. DOM was extracted from forest soils. The processing of this DOM by the ectomycorrhizal fungus Paxillus involutus was examined in laboratory-scale studies with different levels of ammonium. At low levels of ammonium (i.e., under N-limited conditions), the DOM components were slightly oxidized, and fungal C metabolites with iron-reducing activity were secreted. Ammonium amendments decreased the amount of C metabolites, and no additional oxidation of the organic matter was detected. In contrast, the hydrolytic activity and the secretion of N-containing compounds increased, particularly when high levels of ammonium were added. Under these conditions, C, but not N, limited fungal growth. Although the overall production of mineral-associated organic C was not affected by ammonium concentrations, the observed shifts in the activities of the ex vivo and in vivo pathways affected the composition of organic matter adsorbed onto the mineral particles. Such changes will affect the properties of organic matter-mineral associations and, thus, ultimately, the stabilization of SOM.IMPORTANCE Nitrogen (N) availability plays a critical role in the cycling and storage of soil organic matter (SOM). However, large uncertainties remain in predicting the net effect of N addition on soil organic carbon (C) storage due to the complex interactions between organic matter, microbial activity, and mineral particles that determine the formation of stable SOM. Here, we attempted to disentangle the effects of ammonium on these interactions in controlled microcosm experiments including the ectomycorrhizal fungus P.involutus and dissolved organic matter extracted from forest soils. Increased ammonium levels affected the fungal processing of the organic material as well as the secretion of extracellular metabolites. Although ammonium additions did not increase the net production of mineral-adsorbed C, changes in the decomposition and secretion pathways altered the composition of the adsorbed organic matter. These changes may influence the properties of the organic matter-mineral associations and, thus, the stabilization of SOM.

Morphological and ecophysiological root and leaf traits in ectomycorrhizal, arbuscular-mycorrhizal and non-mycorrhizal Alnus incana seedlings

Background and aimsThe aim was to assess possible benefits or drawbacks of arbuscular-mycorrhizal (AM) and ectomycorrhizal (EM) colonisation compared to no mycorrhizas (NM) in seedlings of the same host species. Eight broadleaf species were tested for mycorrhiza formation. Grey alder (Alnus incana) and four fungal species were selected for further experiments.MethodsGrey alder seedlings were inoculated with AM fungi Rhizophagus intraradices and Glomus hoi or EM fungi Paxillus involutus plus an ascomycete isolated from Alnus roots or mock-inoculated (NM).ResultsEM formed in 70% of root tips and AM in 30% of root length. AM plants were smaller than EM and NM, but their specific root length (SRL) and specific leaf area (SLA) were highest. Net photosynthesis, stomatal conductance and shoot water potential did not differ between treatments. Foliar Ca, K, Mg, Mn, N, P and S concentrations (mg g−1) were highest in AM plants. However, total foliar contents (mg per plant) were lowest in AM plants, except for P, K and Zn.ConclusionsThe larger SRL and SLA suggest more efficient resource usage in AM plants, even though these were smaller than EM and NM plants. Grey alder is proposed as a new model species for comparisons between mycorrhiza types in cold climates.

Fenton reaction facilitates organic nitrogen acquisition by an ectomycorrhizal fungus.

Summary Boreal trees rely on their ectomycorrhizal fungal symbionts to acquire growth‐limiting nutrients, such as nitrogen (N), which mainly occurs as proteins complexed in soil organic matter (SOM). The mechanisms for liberating this N are unclear as ectomycorrhizal fungi have lost many genes encoding lignocellulose‐degrading enzymes present in their saprotrophic ancestors. We hypothesized that hydroxyl radicals (˙ OH), produced by the ectomycorrhizal fungus Paxillus involutus during growth on SOM, are involved in liberating organic N. Paxillus involutus was grown for 7 d on N‐containing or N‐free substrates that represent major organic compounds of SOM. ˙ OH production, ammonium assimilation, and proteolytic activity were measured daily. ˙ OH production was strongly induced when P. involutus switched from ammonium to protein as the main N source. Extracellular proteolytic activity was initiated shortly after the oxidation. Oxidized protein substrates induced higher proteolytic activity than unmodified proteins. Dynamic modeling predicted that ˙ OH production occurs in a burst, regulated mainly by ammonium and ferric iron concentrations.We propose that the production of ˙ OH and extracellular proteolytic enzymes are regulated by similar nutritional signals. Oxidation works in concert with proteolysis, improving N liberation from proteins in SOM. Organic N mining by ectomycorrhizal fungi has, until now, only been attributed to proteolysis.

Involutin is an Fe3+ reductant secreted by the ectomycorrhizal fungus Paxillus involutus during Fenton-based decomposition of organic matter.

Ectomycorrhizal fungi play a key role in mobilizing nutrients embedded in recalcitrant organic matter complexes, thereby increasing nutrient accessibility to the host plant. Recent studies have shown that during the assimilation of nutrients, the ectomycorrhizal fungus Paxillus involutus decomposes organic matter using an oxidative mechanism involving Fenton chemistry (Fe2+ + H2O2 + H+ → Fe3+ + ˙OH + H2O), similar to that of brown rot wood-decaying fungi. In such fungi, secreted metabolites are one of the components that drive one-electron reductions of Fe3+ and O2, generating Fenton chemistry reagents. Here we investigated whether such a mechanism is also implemented by P. involutus during organic matter decomposition. Activity-guided purification was performed to isolate the Fe3+-reducing principle secreted by P. involutus during growth on a maize compost extract. The Fe3+-reducing activity correlated with the presence of one compound. Mass spectrometry and nuclear magnetic resonance (NMR) identified this compound as the diarylcyclopentenone involutin. A major part of the involutin produced by P. involutus during organic matter decomposition was secreted into the medium, and the metabolite was not detected when the fungus was grown on a mineral nutrient medium. We also demonstrated that in the presence of H2O2, involutin has the capacity to drive an in vitro Fenton reaction via Fe3+ reduction. Our results show that the mechanism for the reduction of Fe3+ and the generation of hydroxyl radicals via Fenton chemistry by ectomycorrhizal fungi during organic matter decomposition is similar to that employed by the evolutionarily related brown rot saprotrophs during wood decay.

The response of Betula pubescens to inoculation with an ectomycorrhizal fungus and a plant growth promoting bacterium is substrate-dependent

The combination of ectomycorrhizal (ECM) fungi and plant growth promoting bacteria (PGPB) has considerable potential in forestry. Here we report on the combined effects of PGPB Mesorhizobium sp. and the ECM fungus Paxillus involutus on the growth of Betula pubescens in two different substrates, a forest soil and an alkaline anthropogenic sediment. Growth, nutrient concentration and mycorrhizal formation of B. pubescens were determined at the end of the experiment and the fungal and bacterial communities were assessed by denaturing gradient gel electrophoresis. The combined effects of ECM and PGPB enhanced root and shoot growth most in the forest soil with Mesorhizobium appearing to stimulate mycorrhizal formation. However, in the alkaline sediment, an antagonistic interaction was detected between P. involutus and Mesorhizobium sp., with Mesorhizobium sp. alone stimulating growth. The significant interaction between the ECM and the PGBP across the two substrates is discussed.

The carbon starvation response of the ectomycorrhizal fungus Paxillus involutus.

The amounts of carbon allocated to the fungal partner in ectomycorrhizal associations can vary substantially depending on the plant growth and the soil nutrient conditions, and the fungus may frequently be confronted with limitations in carbon. We used chemical analysis and transcriptome profiling to examine the physiological response of the ectomycorrhizal fungus Paxillus involutus to carbon starvation during axenic cultivation. Carbon starvation induced a decrease in the biomass. Concomitantly, ammonium, cell wall material (chitin) and proteolytic enzymes were released into the medium, which suggest autolysis. Compared with the transcriptome of actively growing hyphae, about 45% of the transcripts analyzed were differentially regulated during C-starvation. Induced during starvation were transcripts encoding extracellular enzymes such as peptidases, chitinases and laccases. In parallel, transcripts of N-transporters were upregulated, which suggest that some of the released nitrogen compounds were re-assimilated by the mycelium. The observed changes suggest that the carbon starvation response in P. involutus is associated with complex cellular changes that involves autolysis, recycling of intracellular compounds by autophagy and reabsorption of the extracellular released material. The study provides molecular markers that can be used to examine the role of autolysis for the turnover and survival of the ectomycorrhizal mycelium in soils.

Poplar nutrition under drought as affected by ectomycorrhizal colonization

The nutrient status and physiological responses to drought were investigated in young poplar (Populus×canescens) trees, which were either non-mycorrhizal (NM) or colonized with the ectomycorrhizal fungus Paxillus involutus (EM). Stomatal conductance declined rapidly with limited water availability indicating that P.×canescens is a strongly water-saving tree species. EM trees showed higher stomatal conductance than NM trees but the magnitude of the effect was very small. Nitrogen uptake was traced by 15N. 15N enrichment was higher in EM root tips than in NM root tips. The 15N enrichment in root tips and in roots was significantly linked with stomatal conductance, whereas the enrichment in leaves was correlated with the enrichment in roots, indicating that N uptake is influenced by photosynthetic processes and that the internal translocation to leaves depends on the concentration in the below-ground tissues. Phosphorus and cations, in particular the main osmolyte potassium K, were enriched leaves of EM plants, but not in roots. Under drought, K and Mg increased in roots regardless the EM status, whereas the foliar concentrations increased only in EM plants. A decline in the leaf water content was prevented by increases in both, cations and soluble sugars. EM also delayed root tip mortality compared with NM plants, thus, suggesting transiently positive effects of EM on poplar performance under drought. Under stronger stress at water potentials of about −1.1MPa, the transcript levels of stress marker genes (aquaporin PIP2.5, ABA-responsive RD26, ammonium transporter AMT3.1) were increased regardless the mycorrhizal status of the trees.

Carbon availability triggers the decomposition of plant litter and assimilation of nitrogen by an ectomycorrhizal fungus.

The majority of nitrogen in forest soils is found in organic matter–protein complexes. Ectomycorrhizal fungi (EMF) are thought to have a key role in decomposing and mobilizing nitrogen from such complexes. However, little is known about the mechanisms governing these processes, how they are regulated by the carbon in the host plant and the availability of more easily available forms of nitrogen sources. Here we used spectroscopic analyses and transcriptome profiling to examine how the presence or absence of glucose and/or ammonium regulates decomposition of litter material and nitrogen mobilization by the ectomycorrhizal fungus Paxillus involutus. We found that the assimilation of nitrogen and the decomposition of the litter material are triggered by the addition of glucose. Glucose addition also resulted in upregulation of the expression of genes encoding enzymes involved in oxidative degradation of polysaccharides and polyphenols, peptidases, nitrogen transporters and enzymes in pathways of the nitrogen and carbon metabolism. In contrast, the addition of ammonium to organic matter had relatively minor effects on the expression of transcripts and the decomposition of litter material, occurring only when glucose was present. On the basis of spectroscopic analyses, three major types of chemical modifications of the litter material were observed, each correlated with the expression of specific sets of genes encoding extracellular enzymes. Our data suggest that the expression of the decomposition and nitrogen assimilation processes of EMF can be tightly regulated by the host carbon supply and that the availability of inorganic nitrogen as such has limited effects on saprotrophic activities.

RESISTANCE OF YOUNG STRESSES CAUSED BY HEAVY METALS SUCH AS Cs AND Cd

Using lab experiments, the heavy metal tolerance of young ectomycorrhizal trees was assessed in two axenic systems, co-cultures on germination medium and a sandwich technique system with Modified Melin-Nokrans medium. Sterilized seeds of the tree species Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) were inoculated with the ectomycorrhizal fungi Paxillus involutus and/or Tricholoma vaccinum in media containing either 10 mM CsCl or 0.25 mM CdCl2. The behavior of the seedlings and young trees was investigated over a period of eight weeks. The young mycorrhizal trees showed an increased biomass in comparison to non-mycorrhizal trees. The heavy metals induced stress was visible in reduced germination capacity, reddish colored, instable stems, reduced growth and increased lethality during the first six weeks of growth which was clearly relieved in mycorrhizal association. The results may be important for remediation actions and re-forestation programs.

The ectomycorrhizal fungus Paxillus involutus converts organic matter in plant litter using a trimmed brown-rot mechanism involving Fenton chemistry.

Soils in boreal forests contain large stocks of carbon. Plants are the main source of this carbon through tissue residues and root exudates. A major part of the exudates are allocated to symbiotic ectomycorrhizal fungi. In return, the plant receives nutrients, in particular nitrogen from the mycorrhizal fungi. To capture the nitrogen, the fungi must at least partly disrupt the recalcitrant organic matter–protein complexes within which the nitrogen is embedded. This disruption process is poorly characterized. We used spectroscopic analyses and transcriptome profiling to examine the mechanism by which the ectomycorrhizal fungus Paxillus involutus degrades organic matter when acquiring nitrogen from plant litter. The fungus partially degraded polysaccharides and modified the structure of polyphenols. The observed chemical changes were consistent with a hydroxyl radical attack, involving Fenton chemistry similar to that of brown-rot fungi. The set of enzymes expressed by Pa. involutus during the degradation of the organic matter was similar to the set of enzymes involved in the oxidative degradation of wood by brown-rot fungi. However, Pa. involutus lacked transcripts encoding extracellular enzymes needed for metabolizing the released carbon. The saprotrophic activity has been reduced to a radical-based biodegradation system that can efficiently disrupt the organic matter–protein complexes and thereby mobilize the entrapped nutrients. We suggest that the released carbon then becomes available for further degradation and assimilation by commensal microbes, and that these activities have been lost in ectomycorrhizal fungi as an adaptation to symbiotic growth on host photosynthate. The interdependence of ectomycorrhizal symbionts and saprophytic microbes would provide a key link in the turnover of nutrients and carbon in forest ecosystems.

Nitrous oxide production by the ectomycorrhizal fungi Paxillus involutus and Tylospora fibrillosa

Nitrous oxide (N(2)O) production by filamentous fungi has been demonstrated in pure culture and has been estimated indirectly in soils. However, it is unknown whether ectomycorrhizal fungi can also produce N(2)O. We demonstrate for the first time the ability of nitrogen (N)-tolerant ectomycorrhizal fungi (Paxillus involutus and Tylospora fibrillosa), found in forest soils under moderate to high rates of N deposition, to produce N(2)O from nitrate reduction. The N(2)O concentrations from the ectomycorrhizal fungal treatments after a 10-day pure culture experiment were 0.0117±0.00015 (P. involutus) and 0.0114±0.0003 (T. fibrillosa), and 0.0114±0.00043 μmol N(2)O L(-1) from a known fungal denitrifier (Fusarium lichenicola). No N(2)O was detected in the control treatment. Our results indicate the potential for these two N-tolerant ectomycorrhizal fungi to contribute to N(2)O production. Given that these species are abundant in many forest soils, the strength and regulation of fungal N(2)O production should now be verified in situ.

Ectomycorrhiza formation and willow growth promotion as affected by associated bacteria: role of microbial metabolites and use of C sources

Biotic interactions affect the impact of potential plant growth promoting microorganisms in the rhizosphere, but their magnitude and fundamentals are often hardly known. In a pot experiment, two physiologically different strains of the ectomycorrhizal fungus Paxillus involutus (GUL and FRA) were tested separately and in combination with associated bacteria (Sphingomonas paucimobilis 1 L, Ralstonia pickettii 16 B, Sphingomonas sp. 23 L) on their effects on the growth of willows (Salix viminalis). Both P. involutus strains significantly increased the growth of the willows compared with non-inoculated plants, but the magnitude of this effect was significantly affected by the fungal strain. The P. involutus strain GUL with higher synthesis of auxin-like substances, acid phosphatases, siderophores and faster utilisation of yeast extract in vitro increased the willow growth in situ more effectively than strain FRA. Additionally, dual inoculation with the P. involutus strains GUL and FRA in combination with the associated bacteria promoted the willow growth, especially the combination with S. paucimobilis 1 L. This bacterial strain used effectively C sources which are common components of plant root exudates, e.g. glucose, sucrose, maltose and mannose as well as compounds synthesized by fungi, e.g. trehalose in vitro. We conclude that the analyses of fungal metabolites and of C source use of associated bacteria can successfully contribute to accelerate the selection of capable plant growth promoting combinations.

Upgrading root physiology for stress tolerance by ectomycorrhizas: insights from metabolite and transcriptional profiling into reprogramming for stress anticipation.

Ectomycorrhizas (EMs) alleviate stress tolerance of host plants, but the underlying molecular mechanisms are unknown. To elucidate the basis of EM-induced physiological changes and their involvement in stress adaptation, we investigated metabolic and transcriptional profiles in EM and non-EM roots of gray poplar (Populus x canescens) in the presence and absence of osmotic stress imposed by excess salinity. Colonization with the ectomycorrhizal fungus Paxillus involutus increased root cell volumes, a response associated with carbohydrate accumulation. The stress-related hormones abscisic acid and salicylic acid were increased, whereas jasmonic acid and auxin were decreased in EM compared with non-EM roots. Auxin-responsive reporter plants showed that auxin decreased in the vascular system. The phytohormone changes in EMs are in contrast to those in arbuscular mycorrhizas, suggesting that EMs and arbuscular mycorrhizas recruit different signaling pathways to influence plant stress responses. Transcriptome analyses on a whole genome poplar microarray revealed activation of genes related to abiotic and biotic stress responses as well as of genes involved in vesicle trafficking and suppression of auxin-related pathways. Comparative transcriptome analysis indicated EM-related genes whose transcript abundances were independent of salt stress and a set of salt stress-related genes that were common to EM non-salt-stressed and non-EM salt-stressed plants. Salt-exposed EM roots showed stronger accumulation of myoinositol, abscisic acid, and salicylic acid and higher K(+)-to-Na(+) ratio than stressed non-EM roots. In conclusion, EMs activated stress-related genes and signaling pathways, apparently leading to priming of pathways conferring abiotic stress tolerance.

Influence of ectomycorrhizal colonization on the growth and mineral nutrition of Populus deltoides under Aluminum toxicity

Aluminum (Al) toxicity has been considered an important factor in limiting the growth and nutrient acquisition of sensitive tree species in acidic soils. Mycorrhizal fungi may offset the negative impacts of Al in the root zone. Here, we report our studies on the effect of Al on the growth and mineral nutrition of Populus deltoides in the presence of the ectomycorrhizal fungus Paxillus involutus. Mycorrhizal and non-mycorrhizal plants were exposed to Al levels of 0, 50, 100 and 200 mg/l for 10 weeks. The biomass of mycorrhizal plants increased significantly than non-mycorrhizal plants. The mycorrhizal plants showed higher levels of mineral nutrients such as phosphorus, calcium and magnesium compared to nonmycorrhizal plants in different concentrations of Al. Al content significantly decreased in shoots of mycorrhizal plants compared to non-mycorrhizal plants. The oxalic acid concentration was significantly increased in mycorrhizal plants over non-mycorrhizal plants. These results suggest that ectomycorrhizal colonization confer Al tolerance to P. deltoides plants and Al induced enhancement of organic acids by P. involutus is very likely to be associated with Al tolerance.

Variation in host specificity and gene content in strains from genetically isolated lineages of the ectomycorrhizal fungus Paxillus involutus s. lat.

Ectomycorrhizal fungi are known to vary in host range. Some fungi can enter into symbiosis with multiple plant species, while others have restricted host ranges. The aim of this study was to examine variation in host specificity among strains from the basidiomycete Paxillus involutus s. lat. Recent studies have shown that this fungus consists of at least four genetically isolated lineages, phylogenetic species (PS) I (which corresponds to the morphological species Paxillus obscurosporus), PS II (P. involutus s. str.), PS III (Paxillus validus), and PS IV (not yet supported by any reference material). Thirty-five Paxillus strains of PS I to IV were examined in microcosms for their capacity to infect birch (Betula pendula) and spruce (Picea abies). Seventeen strains were compatible and formed mycorrhizae with both tree species. Seven strains were incompatible with both birch and spruce. The gene content in three pairs of incompatible and compatible strains PS I, II, and III were compared using microarray-based comparative genomic hybridizations. Of 4,113 P. involutus gene representatives analyzed, 390 varied in copy numbers in at least one of the three pairwise comparisons. Only three reporters showed significant changes in all three pairwise comparisons, and none of these were changed in a similar way in three comparisons. Our data indicate that changes in host range have occurred frequently and independently among strains in P. obscurosporus, P. involutus s. str., and P. validus. No evidence was obtained demonstrating that these changes have been associated with the gain or loss of similar genes in these three species.

Phenolic compounds in ectomycorrhizal interaction of lignin modified silver birch.

BackgroundThe monolignol biosynthetic pathway interconnects with the biosynthesis of other secondary phenolic metabolites, such as cinnamic acid derivatives, flavonoids and condensed tannins. The objective of this study is to evaluate whether genetic modification of the monolignol pathway in silver birch (Betula pendula Roth.) would alter the metabolism of these phenolic compounds and how such alterations, if exist, would affect the ectomycorrhizal symbiosis.ResultsSilver birch lines expressing quaking aspen (Populus tremuloides L.) caffeate/5-hydroxyferulate O-methyltransferase (PtCOMT) under the 35S cauliflower mosaic virus (CaMV) promoter showed a reduction in the relative expression of a putative silver birch COMT (BpCOMT) gene and, consequently, a decrease in the lignin syringyl/guaiacyl composition ratio. Alterations were also detected in concentrations of certain phenolic compounds. All PtCOMT silver birch lines produced normal ectomycorrhizas with the ectomycorrhizal fungus Paxillus involutus (Batsch: Fr.), and the formation of symbiosis enhanced the growth of the transgenic plants.ConclusionThe down-regulation of BpCOMT in the 35S-PtCOMT lines caused a reduction in the syringyl/guaiacyl ratio of lignin, but no significant effect was seen in the composition or quantity of phenolic compounds that would have been caused by the expression of PtCOMT under the 35S or UbB1 promoter. Moreover, the detected alterations in the composition of lignin and secondary phenolic compounds had no effect on the interaction between silver birch and P. involutus.