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Andrea Polle - One of the best experts on this subject based on the ideXlab platform.
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ectomycorrhizas with Paxillus involutus enhance cadmium uptake and tolerance in populus canescens
Plant Cell and Environment, 2014Co-Authors: Jiali He, Andrea Polle, Hong Li, Changhui PengAbstract:: Ectomycorrhizas (EMs), which are symbiotic organs formed between tree roots and certain fungi, can mediate cadmium (Cd) tolerance of host plants, but the underlying physiological and molecular mechanisms are not fully understood. To investigate EMs mediated Cd tolerance in woody plants, Populus × canescens was inoculated with Paxillus involutus (strain MAJ) to establish mycorrhizal roots. Mycorrhizal poplars and non-mycorrhizal controls were exposed to 0 or 50 μM CdSO4 . EMs displayed higher net Cd(2+) influx than non-mycorrhizal roots. Net Cd(2+) influx was coupled with net H(+) efflux and inactivation of plasma membrane (PM) H(+) -ATPases reduced Cd(2+) uptake of EMs less than of non-mycorrhizal roots. Consistent with higher Cd(2+) uptake in EMs, in most cases, transcript levels of genes involved in Cd(2+) uptake, transport and detoxification processes were increased in EMs compared to non-mycorrhizal roots. Higher CO2 assimilation, improved nutrient and carbohydrate status, and alleviated oxidative stress were found in mycorrhizal compared to non-mycorrhizal poplars despite higher Cd(2+) accumulation. These results indicate that mycorrhizas increase Cd(2+) uptake, probably by an enlarged root volume and overexpression of genes involved in Cd(2+) uptake and transport, and concurrently enhance Po. × canescens Cd tolerance by increased detoxification, improved nutrient and carbohydrate status and defence preparedness.
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Ectomycorrhizas with Paxillus involutus enhance cadmium uptake and tolerance in Populus × canescens
Plant Cell and Environment, 2013Co-Authors: Yonglu Ma, Andrea Polle, Jiali He, Hong Li, Chaofeng Ma, Changhui PengAbstract:: Ectomycorrhizas (EMs), which are symbiotic organs formed between tree roots and certain fungi, can mediate cadmium (Cd) tolerance of host plants, but the underlying physiological and molecular mechanisms are not fully understood. To investigate EMs mediated Cd tolerance in woody plants, Populus × canescens was inoculated with Paxillus involutus (strain MAJ) to establish mycorrhizal roots. Mycorrhizal poplars and non-mycorrhizal controls were exposed to 0 or 50 μM CdSO4 . EMs displayed higher net Cd(2+) influx than non-mycorrhizal roots. Net Cd(2+) influx was coupled with net H(+) efflux and inactivation of plasma membrane (PM) H(+) -ATPases reduced Cd(2+) uptake of EMs less than of non-mycorrhizal roots. Consistent with higher Cd(2+) uptake in EMs, in most cases, transcript levels of genes involved in Cd(2+) uptake, transport and detoxification processes were increased in EMs compared to non-mycorrhizal roots. Higher CO2 assimilation, improved nutrient and carbohydrate status, and alleviated oxidative stress were found in mycorrhizal compared to non-mycorrhizal poplars despite higher Cd(2+) accumulation. These results indicate that mycorrhizas increase Cd(2+) uptake, probably by an enlarged root volume and overexpression of genes involved in Cd(2+) uptake and transport, and concurrently enhance Po. × canescens Cd tolerance by increased detoxification, improved nutrient and carbohydrate status and defence preparedness.
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the ectomycorrhizal fungus Paxillus involutus modulates leaf physiology of poplar towards improved salt tolerance
Environmental and Experimental Botany, 2011Co-Authors: Ke Li, Xiangning Jiang, Cornelia Gobel, Henning Wildhagen, Ivo Feusner, Heinz Rennenberg, Andrea PolleAbstract:Abstract Ectomycorrhizas (EMs) are mutualistic associations between soil fungi and plant roots. Although the physical interaction occurs only in roots, mycorrhizas may alter the physiology of the whole plant, resulting in changes in host responses to abiotic stress. To elucidate the influence of an ectomycorrhizal fungus on leaf physiology and performance under salt stress, we analysed the levels of nutrient elements, phytohormones, carbohydrates, amino compounds and fatty acids in leaves of Populus × canescens. The poplars were cultivated either in the presence or absence of Paxillus involutus and either with or without salt stress imposed by 150 mM NaCl. Leaves of ectomycorrhizal plants displayed higher quantum yield of photochemistry (ΦPSII), increased concentrations of phosphorus and potassium, decreased concentrations of galactose, increased concentrations of the stress metabolite γ-amino butyric acid and a lower unsaturated-to-saturated fatty acid ratios than those of non-ectomycorrhizal plants. Salt exposure of P. × canescens led to leaf chlorosis and shedding, decreases in ΦPSII, K+-to-Na+ ratio, 9Z-hexadecenoic acid, 9Z-octadecenoic acid and unsaturated-to-saturated fatty acid ratio, and increases in ABA, glucose, fructose and some amino compounds. Under salinity leaves of ectomycorrhizal plants showed an alleviation of leaf chlorosis, improved water status, higher ΦPSII and K+-to-Na+ ratio, less accumulation of major amino compounds and lower unsaturated-to-saturated fatty acid ratios than those of non-mycorrhizal plants. These results indicate that ectomycorrhizas attenuate salinity induced injury in leaves of P. × canescens, which may be ascribed to the improved nutrient status, osmo-regulation and changes in fatty acid composition in leaves of ectomycorrhizal plants.
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ectomycorrhizal fungus Paxillus involutus and hydrogels affect performance of populus euphratica exposed to drought stress
Annals of Forest Science, 2009Co-Authors: Ke Li, Xiangning Jiang, Andrea PolleAbstract:• Mycorrhizal fungi and hydrogels (water-absorbing polymers) can improve water availability for trees. The combination of both factors for plant performance under water limitation has not yet been studied. • To investigate the influence of the ectomycorrhizal fungus Paxillus involutus, hydrogel and the combination of both factors, a drought-sensitive poplar, Populus euphratica, was examined in this study. • After 16 weeks of inoculation, no ectomycorrhizas were found. Nevertheless, P. involutus-inoculated poplars displayed increased concentrations of soluble sugars and osmolality, leading to an improved water status. Growth was diminished compared with non-inoculated P. euphratica. The presence of hydrogels in the rooting medium resulted in increased biomass and higher plant water content and decreased the osmolality of plant tissues. Drought markedly decreased water contents in rooting medium and plants, and leaf chlorophyll fluorescence, and stimulated the root growth, concentrations of soluble sugars and osmolality in plants. Under drought conditions, P. euphratica exhibited osmoregulation by accumulation of low-molecular-weight carbohydrates. • These data indicate that adding hydrogels to soils may improve the plant performance. The reasons for improved osmoregulation by fungi and hydrogels were probably related to their stimulating influence on the formation of soluble carbohydrates under drought conditions.
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peroxidase activity in poplar inoculated with compatible and incompetent isolates of Paxillus involutus
Hayati Journal of Biosciences, 2007Co-Authors: Abdul Gafur, Andres Schutzendubel, Andrea PolleAbstract:Peroxidase activity of the hybrid poplar Populus x canescens (Ait.) Sm. (= P. tremula L. x P. alba L.) inoculated with compatible and incompetent isolates of Paxillus involutus (Batsch) Fr. was investigated. Screening of the ectomycorrhizal fungal isolates was initiated with exploration of mycelial growth characteristics and mycorrhizal ability in vitro with poplar. Both traits varied within the fungus although they did not seem to be genetically correlated. While isolates SCO1, NAU, and 031 grew faster than others, only isolates MAJ, SCO1, and 031 were able to form ectomycorrhiza with poplar. Isolates MAJ (compatible) and NAU (incompetent) were subsequently selected for further experiments. Activity of peroxidase, one of the defense-related enzymes, was examined in pure culture and short root components of compatible and incompetent interactions between poplar and P. involutus. Peroxidase activities increased significantly in poplar inoculated with incompetent isolate of the fungus compared to control, while induction of the same enzyme was not detected in compatible associations. Key words: ectomycorrhiza, Paxillus involutus, peroxidase, plant defense, poplar
Michel Chalot - One of the best experts on this subject based on the ideXlab platform.
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Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus.
Microbiology (Reading England), 2020Co-Authors: Damien Blaudez, Bernard Botton, Michel ChalotAbstract:Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus were investigated using radiotracer flux analyses. Concentration-dependent Cd2+-uptake kinetics were characterized by a smooth, non-saturating curve that could be dissected into linear and saturable components. The linear-uptake kinetic component was interpreted as representing binding of Cd to apoplastic components, whereas the remaining saturable component was the result of carrier-mediated transport across the plasma membrane. Cell-wall-bound Cd was almost completely removed during desorption from cell-wall preparations. Cd2+ desorption from intact mycelium was found to be a function of time involving three compartments corresponding in series to cell wall (50%), cytoplasm (30%) and vacuole (20%), when mycelia were exposed to a 0.05 microM Cd concentration. At 4 degrees C, most of the Cd recovered was due to the cell-wall-bound fraction, suggesting that transport across the plasma membrane is a metabolically mediated process. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited Cd accumulation in P. involutus mycelia by up to 28%, which indicates that transport of Cd2+ was partially dependent on the membrane potential. Cd2+ uptake into symplasm is linked to Ca2+ transport, as revealed by the inhibition of Cd accumulation by the Ca2+ ionophore A23187. The present work demonstrates the ability of the ectomycorrhizal fungus P. involutus to take up and further accumulate Cd in different compartments. Binding of Cd onto cell walls and accumulation of Cd in the vacuolar compartment may be regarded as two essential metal-detoxification mechanisms. These data represent a first step towards the understanding of metal-tolerance mechanisms in mycorrhizal fungi.
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characterization and regulation of pidur3 a permease involved in the acquisition of urea by the ectomycorrhizal fungus Paxillus involutus
Fungal Genetics and Biology, 2008Co-Authors: Melanie Morel, Michel Chalot, Christophe Jacob, Michael Fitz, Daniel Wipf, Annick BrunAbstract:Abstract Urea, which is known to be a source of nitrogen for the growth of many organisms, represents an important fertilizer in forest soils. Since most trees form symbiotic associations with ectomycorrhizal fungi, the capacities of these symbionts to take up and assimilate urea would determine the efficiency of urea nitrogen salvaging by plants. We showed that Paxillus involutus, an ectomycorrhizal basidiomycete, is capable of using urea as sole nitrogen source. We report the molecular characterization of an active urea transporter (PiDur3) isolated from this fungus. We demonstrated that the import of urea is a minor event on ammonium condition, since the expression of PiDUR3 is repressed by the high intracellular glutamine pool. Interestingly, on urea nutritive condition, the uptake of urea is rather mediated by the intracellular urea pool and particularly by urease efficiency.
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metal induction of a Paxillus involutus metallothionein and its heterologous expression in hebeloma cylindrosporum
New Phytologist, 2007Co-Authors: Marc Bellion, Mikael Courbot, Christophe Jacob, Frederic Guinet, Damien Blaudez, Michel ChalotAbstract:Summary • Metallothioneins are small polypeptides involved in metal tolerance of many eukaryotes. Here we characterized the Pimt1 gene, coding for a metallothionein from the ectomycorrhizal fungus Paxillus involutus. • Expression of Pimt1 in P. involutus under metal stress conditions was measured by northern blot and RT-PCR analyses. The full-length cDNA was used to perform functional complementation in yeast mutant strains and agrotransformation of Hebeloma cylindrosporum. • Heterologous expression in yeast showed that PiMT1 was able to complement the hypersensitivity of mutant strains to cadmium (Cd) and copper (Cu), but not to zinc (Zn). Transcripts were almost undetectable under control conditions, whereas Cu and Cd, but not Zn, strongly induced Pimt1 expression in P. involutus. Constitutive overexpression of Pimt1 in H. cylindrosporum conferred a higher copper tolerance. • The present study identified PiMT1 as a potential determinant in the response of mycorrhizal fungi to Cu and Cd stress. Additionally, we demonstrated the usefulness of mycorrhizal fungi transformation using Agrobacterium technology to approach gene function.
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identification of genes differentially expressed in extraradical mycelium and ectomycorrhizal roots during Paxillus involutus betula pendula ectomycorrhizal symbiosis
Applied and Environmental Microbiology, 2005Co-Authors: Ma â â Lanie Morel, Michel Chalot, Christophe Jacob, Francis Martin, Tomas Johansson, Annegret Kohler, Annick BrunAbstract:The development of ectomycorrhizal symbiosis leads to drastic changes in gene expression in both partners. However, little is known about the spatial regulation of symbiosis-regulated genes. Using cDNA array profiling, we compared the levels of expression of fungal genes corresponding to approximately 1,200 expressed sequenced tags in the ectomycorrhizal root tips (ECM) and the connected extraradical mycelium (EM) for the Paxillus involutus-Betula pendula ectomycorrhizal association grown on peat in a microcosm system. Sixty-five unique genes were found to be differentially expressed in these two fungal compartments. In ECM, a gene coding for a putative phosphatidylserine decarboxylase (Psd) was up-regulated by 24-fold, while genes coding for urea (Dur3) and spermine (Tpo3) transporters were up-regulated 4.1- and 6.2-fold in EM. Moreover, urea was the major nitrogen compound found in EM by gas chromatography-mass spectrometry analysis. These results suggest that (i) there is a spatial difference in the patterns of fungal gene expression between ECM and EM, (ii) urea and polyamine transporters could facilitate the translocation of nitrogen compounds within the EM network, and (iii) fungal Psd may contribute to membrane remodeling during ectomycorrhiza formation.
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cadmium responsive thiols in the ectomycorrhizal fungus Paxillus involutus
Applied and Environmental Microbiology, 2004Co-Authors: Mikael Courbot, Michel Chalot, Laurent Diez, Roberta Ruotolo, Pierre LeroyAbstract:Molecular and cellular mechanisms underlying the sustained metal tolerance of ectomycorrhizal fungi are largely unknown. Some of the main mechanisms involved in metal detoxification appear to involve the chelation of metal ions in the cytosol with thiol-containing compounds, such as glutathione, phytochelatins, or metallothioneins. We used an improved high-performance liquid chromatography method for the simultaneous measurement of thiol-containing compounds from cysteine and its derivatives (γ-glutamylcysteine, glutathione) to higher-molecular-mass compounds (phytochelatins). We found that glutathione and γ-glutamylcysteine contents increased when the ectomycorrhizal fungus Paxillus involutus was exposed to cadmium. An additional compound with a 3-kDa molecular mass, most probably related to a metallothionein, increased drastically in mycelia exposed to cadmium. The relative lack of phytochelatins and the presence of a putative metallothionein suggest that ectomycorrhizal fungi may use a different means to tolerate heavy metals, such as Cd, than do their plant hosts.
Georg Jentschke - One of the best experts on this subject based on the ideXlab platform.
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interdependence of phosphorus nitrogen potassium and magnesium translocation by the ectomycorrhizal fungus Paxillus involutus
New Phytologist, 2001Co-Authors: Georg Jentschke, Bettina Brandes, A J Kuhn, Walter H Schroder, Douglas L GodboldAbstract:• Translocation is shown of phosphorus, nitrogen, potassium and magnesium to a P-deficient host from ectomycorrhizal fungal hyphae. • Mycorrhizal (with Paxillus involutus) and nonmycorrhizal P-deficient spruce (P. abies) seedlings were grown in a two-compartment sand-culture system. Hyphal translocation of nutrients from the inner compartment (penetrated only by hyphae) to the host was measured using mass balance (for N, P and K) or stable isotope (15N and 25Mg) methods. • Addition of P to the hyphal compartment strongly stimulated hyphal growth, and this also increased both seedling P status and growth. Hyphae translocated nonlimiting elements in addition to P, contributing 52, 17, 5 and 3–4%, respectively, to total P, N, K or Mg plant uptake. The potential role of the ectomycorrhizal mycelium in K acquisition was demonstrated. Translocation to mycorrhizal seedings of N, K and Mg was strongly reduced when hyphal P-fluxes ceased; this translocation of nonlimiting nutrients depended on simultaneous translocation of P. • The ectomycorrhizal mycelium has an active role in P acquisition from sources not available to roots. Nutrient fluxes within fungal hyphae are interdependent and strong coupling of N, K and Mg fluxes with long-distance P translocation in the mycorrhizal mycelium occurs.
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the mycorrhizal fungus Paxillus involutus transports magnesium to norway spruce seedlings evidence from stable isotope labeling
Plant and Soil, 2000Co-Authors: Georg Jentschke, Bettina Brandes, A J Kuhn, Walter H Schroder, J S Becker, Douglas L GodboldAbstract:Although it is well established that ectomycorrhizas improve the mineral nutrition of forest trees, there has been little evidence that they mediate uptake of divalent cations such as Mg. We grew nonmycorrhizal seedlings and seedlings mycorrhizal with Paxillus involutus Batsch in a sand culture system with two compartments separated by a 45-μm Nylon mesh. Hyphae, but not roots, can penetrate this net. Labeling the compartment only accessible to hyphae with 25Mg showed that hyphae of the ectomycorrhizal fungus Paxillus involutus transported Mg to their host plant. No label was found in nonmycorrhizal control plants. Our data support the idea that ectomycorrhizas are important for the Mg nutrition of forest trees.
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nitrogen and phosphorus acquisition by the mycelium of the ectomycorrhizal fungus Paxillus involutus and its effect on host nutrition
New Phytologist, 1998Co-Authors: Bettina Brandes, Georg Jentschke, A J Kuhn, Douglas L GodboldAbstract:summary The contribution of the extramatrical mycelium to N and P nutrition of mycorrhizal Norway spruce (Picea abies (L.) Karst.) was investigated. Seedlings either inoculated with Paxillus involutus (Batsch) Fr. or non-mycorrhizal were grown in a two compartment sand culture system where hyphae were separated from roots by a 45 lm nylon net. Nutrient solution of the hyphal compartment contained either 1‐ 8m m NH % + and 0‐18 mm H # PO % ’ or no N and P. Aluminium added to the hyphal compartment as a tracer of mass flow was not detected in the plant compartment, indicating that measurements of N and P transfer by the mycelium were not biased by solute movement across the nylon net. The addition of N and P to the hyphal compartment markedly increased dry weight, N and P concentration and N and P content of mycorrhizal plants. Calculating uptake from the dierence in input and output of nutrient in solution confirmed a hyphal contribution of 73% and 76% to total N and P uptake, respectively. Hyphal growth was increased at the site of nutrient solution input.
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the effect of Paxillus involutus fr on aluminum sensitivity of norway spruce seedlings
Tree Physiology, 1993Co-Authors: Eike Hentschel, Douglas L Godbold, Petra Marschner, Hedda Schlegel, Georg JentschkeAbstract:: Non-mycorrhizal Norway spruce seedlings (Picea abies Karst.) and Norway spruce seedlings colonized with Paxillus involutus Fr. were grown in an axenic silica sand culture system. After successful mycorrhizal colonization, the seedlings were exposed to 200 or 800 micro M AlCl(3) for 10 weeks. In both non-mycorrhizal and mycorrhizal seedlings, exposure to Al significantly reduced root growth and the uptake of Mg and Ca. After 5 weeks of exposure to 800 micro M Al, the mycorrhizal seedlings had significantly higher chlorophyll concentrations than the non-mycorrhizal seedlings, although no difference in Mg nutrition was apparent. After 10 weeks of exposure to Al, both non-mycorrhizal and mycorrhizal seedlings exhibited needle chlorosis and reduced photosynthetic activity. However, the aluminum-induced reduction in shoot growth was largely ameliorated by colonization with P. involutus. We conclude that mycorrhizal colonization modifies the phytotoxic effects of Al in Norway spruce seedlings. However, differences in physiological responses to Al between mycorrhizal and non-mycorrhizal seedlings may be largely reduced in the long term as a result of impaired mineral nutrient uptake.
Anders Tunlid - One of the best experts on this subject based on the ideXlab platform.
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three redundant synthetases secure redox active pigment production in the basidiomycete Paxillus involutus
Chemistry & Biology, 2015Co-Authors: Jana Braesel, Sebastian Gotze, Firoz Shah, Daniel Heine, James P Tauber, Christian Hertweck, Anders Tunlid, Pierre Stallforth, Dirk HoffmeisterAbstract:The symbiotic fungus Paxillus involutus serves a critical role in maintaining forest ecosystems, which are carbon sinks of global importance. P. involutus produces involutin and other 2,5-diarylcyclopentenone pigments that presumably assist in the oxidative degradation of lignocellulose via Fenton chemistry. Their precise biosynthetic pathways, however, remain obscure. Using a combination of biochemical, genetic, and transcriptomic analyses, in addition to stable-isotope labeling with synthetic precursors, we show that atromentin is the key intermediate. Atromentin is made by tridomain synthetases of high similarity: InvA1, InvA2, and InvA5. An inactive atromentin synthetase, InvA3, gained activity after a domain swap that replaced its native thioesterase domain with that of InvA5. The found degree of multiplex biosynthetic capacity is unprecedented with fungi, and highlights the great importance of the metabolite for the producer.
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the carbon starvation response of the ectomycorrhizal fungus Paxillus involutus
FEMS Microbiology Ecology, 2015Co-Authors: Magnus Ellstrom, Firoz Shah, Tomas Johansson, Dag Ahren, Per Persson, Anders TunlidAbstract: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.
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the molecular components of the extracellular protein degradation pathways of the ectomycorrhizal fungus Paxillus involutus
New Phytologist, 2013Co-Authors: Firoz Shah, Tomas Johansson, Francois Rineau, Bjorn Canback, Anders TunlidAbstract:Proteins contribute to a major part of the organic nitrogen (N) in forest soils. This N is mobilized and becomes available to trees as a result of the depolymerizing activities of symbiotic ectomycorrhizal fungi. The mechanisms by which these fungi depolymerize proteins and assimilate the released N are poorly characterized. Biochemical analysis and transcriptome profiling were performed to examine the proteolytic machinery and the uptake system of the ectomycorrhizal basidiomycete Paxillus involutus during the assimilation of organic N from various protein sources and extracts of organic matter. All substrates induced secretion of peptidase activity with an acidic pH optimum, mostly contributed by aspartic peptidases. The peptidase activity was transiently repressed by ammonium. Transcriptional analysis revealed a large number of extracellular endo- and exopeptidases. The expression levels of these peptidases were regulated in parallel with transporters and enzymes involved in the assimilation and metabolism of the released peptides and amino acids. For the first time the molecular components of the protein degradation pathways of an ectomycorrhizal fungus are described. The data suggest that the transcripts encoding these components are regulated in response to the chemical properties and the availability of the protein substrates.
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The molecular components of the extracellular protein‐degradation pathways of the ectomycorrhizal fungus Paxillus involutus
New Phytologist, 2013Co-Authors: Firoz Shah, Tomas Johansson, Francois Rineau, Bjorn Canback, Anders TunlidAbstract:Proteins contribute to a major part of the organic nitrogen (N) in forest soils. This N is mobilized and becomes available to trees as a result of the depolymerizing activities of symbiotic ectomycorrhizal fungi. The mechanisms by which these fungi depolymerize proteins and assimilate the released N are poorly characterized. Biochemical analysis and transcriptome profiling were performed to examine the proteolytic machinery and the uptake system of the ectomycorrhizal basidiomycete Paxillus involutus during the assimilation of organic N from various protein sources and extracts of organic matter. All substrates induced secretion of peptidase activity with an acidic pH optimum, mostly contributed by aspartic peptidases. The peptidase activity was transiently repressed by ammonium. Transcriptional analysis revealed a large number of extracellular endo- and exopeptidases. The expression levels of these peptidases were regulated in parallel with transporters and enzymes involved in the assimilation and metabolism of the released peptides and amino acids. For the first time the molecular components of the protein degradation pathways of an ectomycorrhizal fungus are described. The data suggest that the transcripts encoding these components are regulated in response to the chemical properties and the availability of the protein substrates.
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multiple gene genealogies and species recognition in the ectomycorrhizal fungus Paxillus involutus
Fungal Biology, 2008Co-Authors: Jenny Hedh, Peter Samson, Susanne Erland, Anders TunlidAbstract:Paxillus involutus (basidiomycetes, Boletales) is a common ectomycorrhizal fungus in the Northern Hemisphere. The fungus displays significant variation in phenotypic characters related to morphology, physiology, and ecology. Previous studies have shown that P. involutus contains several intersterility groups and morphological species. In this study, we have used concordance of multiple gene genealogies to identify genetically isolated species of P. involutus. Fragments from five protein coding genes in 50 isolates of P. involutus collected from different hosts and environments in Europe and one location in Canada were analysed using phylogenetic methods. Concordance of the five gene genealogies showed that P. involutus comprises at least four distinct phylogenetic lineages: phylogenetic species I (with nine isolates), II (33 isolates), III (three isolates), and IV (five isolates). The branches separating the four species were long and well supported compared with the species internodes. A low level of shared polymorphisms was observed among the four lineages indicating a long time since the genetic isolation began. Three of the phylospecies corresponded to earlier identified morphological species: I to P. obscurosporus, II to P. involutus s. str., and III to P. validus. The phylogenetic species had an overlapping geographical distribution. Species I and II differed partly in habitat and host preferences.
Bernard Botton - One of the best experts on this subject based on the ideXlab platform.
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Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus.
Microbiology (Reading England), 2020Co-Authors: Damien Blaudez, Bernard Botton, Michel ChalotAbstract:Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus were investigated using radiotracer flux analyses. Concentration-dependent Cd2+-uptake kinetics were characterized by a smooth, non-saturating curve that could be dissected into linear and saturable components. The linear-uptake kinetic component was interpreted as representing binding of Cd to apoplastic components, whereas the remaining saturable component was the result of carrier-mediated transport across the plasma membrane. Cell-wall-bound Cd was almost completely removed during desorption from cell-wall preparations. Cd2+ desorption from intact mycelium was found to be a function of time involving three compartments corresponding in series to cell wall (50%), cytoplasm (30%) and vacuole (20%), when mycelia were exposed to a 0.05 microM Cd concentration. At 4 degrees C, most of the Cd recovered was due to the cell-wall-bound fraction, suggesting that transport across the plasma membrane is a metabolically mediated process. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited Cd accumulation in P. involutus mycelia by up to 28%, which indicates that transport of Cd2+ was partially dependent on the membrane potential. Cd2+ uptake into symplasm is linked to Ca2+ transport, as revealed by the inhibition of Cd accumulation by the Ca2+ ionophore A23187. The present work demonstrates the ability of the ectomycorrhizal fungus P. involutus to take up and further accumulate Cd in different compartments. Binding of Cd onto cell walls and accumulation of Cd in the vacuolar compartment may be regarded as two essential metal-detoxification mechanisms. These data represent a first step towards the understanding of metal-tolerance mechanisms in mycorrhizal fungi.
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molecular cloning characterization and regulation by cadmium of a superoxide dismutase from the ectomycorrhizal fungus Paxillus involutus
FEBS Journal, 2001Co-Authors: Christophe Jacob, Annick Brun, Mikael Courbot, Bernard Botton, Howard M Steinman, Jeanpierre Jacquot, Michel ChalotAbstract:The gene encoding a superoxide dismutase (PiSOD) was cloned by suppressive subtractive hybridization from cDNA library of the ectomycorrhizal fungus, Paxillus involutus, grown under cadmium-stress conditions. The encoded protein was presumed to be localized in the peroxisomes because it contained a C-terminal peroxisomal localization peptide (SKL) and lacked an N-terminal mitochondrial transit peptide. Complementation of an Escherichia coli SOD null strain that is unable to grow in the presence of paraquat or cadmium indicated that cloned Pisod encoded a functional superoxide dismutase. Sensitivity of PiSOD activity to H2O2 but not KCN, and sequence homologies to other SODs strongly suggest that it is a manganese-containing superoxide dismutase. Monitoring PiSOD transcript, immunoreactive polypeptide and superoxide dismutase activity following cadmium stress suggests that the principal level of control is post-translational. This is, to our knowledge, the first insight in the characterization of molecular events that take place in an ectomycorrhizal fungus during exposure to heavy metals.
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effects of heavy metals on nitrogen uptake by Paxillus involutus and mycorrhizal birch seedlings
FEMS Microbiology Ecology, 2000Co-Authors: Damien Blaudez, Bernard Botton, Michel ChalotAbstract:The effects of the heavy metals Cu, Cd, Ni, Pb and Zn on [14C]methylamine and [14C]aminoisobutyric acid uptake were studied in the free-living fungus Paxillus involutus and in mycorrhizal and non-mycorrhizal birch roots. The uptake of both N sources by P. involutus was inhibited by the five metals tested. However, Cu2+ and Pb2+ had a greater inhibitory effect. Non-competitive inhibitions were determined between heavy metals and [14C]methylamine uptake. [14C]Methylamine uptake was reduced by one third by 2 μM Cd2+ and Cu2+ in non-mycorrhizal roots, whereas that of mycorrhizal roots was not affected. However, it was reduced by 30 to 80% by 200 μM Cd2+ and Cu2+ irrespective of the mycorrhizal status. [14C]Aminoisobutyric acid uptake in mycorrhizal roots was not significantly affected by Cd2+ and Cu2+, whereas that of non-mycorrhizal roots was decreased by 77% at 200 μM Cu2+. [14C]Aminoisobutyric acid uptake was 4.5 to 6 fold higher in mycorrhizal roots, compared with non-mycorrhizal roots, even under metal exposure. The high efficiency of N acquisition by mycorrhizal birch seedlings under metal exposure might be regarded as a mechanism of stress avoidance.
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cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus
Microbiology, 2000Co-Authors: Damien Blaudez, Bernard Botton, Michel ChalotAbstract:Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus were investigated using radiotracer flux analyses. Concentration-dependent Cd2+-uptake kinetics were characterized by a smooth, non-saturating curve that could be dissected into linear and saturable components. The linear-uptake kinetic component was interpreted as representing binding of Cd to apoplastic components, whereas the remaining saturable component was the result of carrier-mediated transport across the plasma membrane. Cell-wall-bound Cd was almost completely removed during desorption from cell-wall preparations. Cd2+ desorption from intact mycelium was found to be a function of time involving three compartments corresponding in series to cell wall (50%), cytoplasm (30%) and vacuole (20%), when mycelia were exposed to a 0·05 μM Cd concentration. At 4 °C, most of the Cd recovered was due to the cell-wall-bound fraction, suggesting that transport across the plasma membrane is a metabolically mediated process. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited Cd accumulation in P. involutus mycelia by up to 28%, which indicates that transport of Cd2+ was partially dependent on the membrane potential. Cd2+ uptake into symplasm is linked to Ca2+ transport, as revealed by the inhibition of Cd accumulation by the Ca2+ ionophore A23187. The present work demonstrates the ability of the ectomycorrhizal fungus P. involutus to take up and further accumulate Cd in different compartments. Binding of Cd onto cell walls and accumulation of Cd in the vacuolar compartment may be regarded as two essential metal-detoxification mechanisms. These data represent a first step towards the understanding of metal-tolerance mechanisms in mycorrhizal fungi.
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ammonium and methylamine transport by the ectomycorrhizal fungus Paxillus involutus and ectomycorrhizas
FEMS Microbiology Ecology, 1999Co-Authors: Arnaud Javelle, Michel Chalot, Bengt Soderstrom, Bernard BottonAbstract:Using [14C]methylamine as an analogue of ammonium, the kinetics and the energetics of NH4+ transport were studied in the ectomycorrhizal fungus, Paxillus involutus (Batsch) Fr. The apparent half-saturation constant (Km) and the maximum uptake rate (Vmax) for the carrier-mediated transport derived from the Eadie-Hofstee transformation were 180 μM and 380 nmol (mg dry wt)−1 min−1, respectively. Both pH dependence and inhibition by protonophores indicate that methylamine transport in P. involutus was dependent on the electrochemical H+ gradient. Both long-term and short-term uptake experiments were consistent with regulation of ammonium/methylamine transport processes by the presence of an organic nitrogen source. Analysis of methylamine uptake by different P. involutus isolates revealed no obvious trend in the uptake capacities in relation to N deposition at the collection site. Kinetic parameters were determined in P. involutus/Betula pendula (Roth.) axenic association and in detached mycorrhizal roots isolated from forest sites. Enhanced methylamine uptake in the presence of the fungal symbiont was demonstrated. Homogeneous Vmax values were found for axenic and detached mycorrhizas, whereas Km values showed greater variations.