Many were named, but few are current: The Hebeloma of Hesler, Smith, and coauthors.

During the 1970s and 1980s, L. R. Hesler and A. H. Smith, alone, together, or Smith with other authors such as V. S. Evenson and D. H. Mitchel, described numerous North American taxa in Hebeloma. With the inclusion of an early work by Smith and a later work by E. Grilli, who described a species based on material from Smith, 130 taxa were described and form the subject of this paper. Apart from two taxa that were (deliberately) invalidly published and two that were illegitimately published, all others are valid and legitimate names. After study of morphology, habitat, and location of collection (based on available material and information) as well as molecular analysis (insofar as this was successful), of these 128 validly published taxa we regard 14 as being current names; the remaining 114 are synonymized with other current names. These 14 species are Hebeloma albomarginatum, H. alpinicola, H. angelesiense, H. caulocystidiosum, H. immutabile, H. incarnatulum, H. kelloggense, H. mackinawense, H. nitidum, H. olympianum, H. parcivelum, H. praeolidum, H. pungens, and H. sporadicum. This brings up the number of currently recognized, validly published, Hebeloma species in America to 72.

Old but not obsolete: A new life for some of Murrill's (Hebeloma) names.

William Alphonso Murrill was an American mycologist of the early 20th century. He described 1453 new species of Agaricales, Boletales, and Polyporales. Within these were 44 taxa that he described as Hebeloma or that he recombined into Hebeloma. Additionally, there are five species, of which we are aware, that Murrill described within other genera that should be referred to the genus Hebeloma. A further three species described from northern America by J. P. F. C. Montagne, and transferred to Hebeloma by Saccardo, were commented on by Murrill and not accepted within the genus. These 52 taxa are analyzed here, both morphologically and molecularly, as far as possible. For 18 of his types, internal transcribed spacer (ITS) sequences were generated. For two species (H. harperi and H. subfastibile), which were mixed collections, lectotypes are designated. Twenty-three of the taxa analyzed are Hebeloma, as the genus is recognized today, and six of these (H. australe, H. harperi, H. paludicola, H. subaustrale, H. subfastibile, and H. viscidissimum) are regarded as current, i.e., they are names that should be accepted and used. Hebeloma paludicola is an earlier name for H. hygrophilum, described from Europe. Gymnopilus viscidissimus is synonymous with H. amarellum but has priority and is here recombined into Hebeloma. The remaining 17 Hebeloma taxa are synonymized with other species that have priority. The remaining 29 species belong to a range of genera; molecularly supported were Agrocybe, Cortinarius, Inocybe, Inosperma, Phlegmacium, Pholiota, Pseudosperma, and Pyrrhulomyces. Recombinations and synonymizations are made as appropriate and necessary. The names H. alachuanum and H. vatricosum, respectively Inocybe vatricosa, are considered doubtful and should be avoided.

Diverse partitiviruses hosted by the ectomycorrhizal agaric Hebeloma mesophaeum and the natural transmission of a partitivirus between phylogenetically distant, sympatric fungi.

Mycorrhizal fungi host diverse mycoviruses that contribute to our understanding of their diversity and evolution. Here we report on the identification and complete genome characterization of three novel partitiviruses naturally infecting the ectomycorrhizal fungus Hebeloma mesophaeum. During NGS derived viral sequence analyses, we identified a partitivirus that is conspecific with the previously reported partitivirus (LcPV1) described from a saprotrophic fungus Leucocybe candicans. The two distinct fungal specimens inhabited the same vicinity of a campus garden. RdRp sequences encoded by the LcPV1 isolates from both host fungi was found to be identical. Bio-tracking studies revealed that viral loads of LcPV1 drop significantly in L. candicans but not in H. mesophaeum within four years period. The physical proximity of the mycelial networks of both fungal specimens implied the occurrence of a virus transmission event with unknown mechanism. Nature of this virus transmission was discussed in relation to transient interspecific mycelial contact hypothesis.

96 North American taxa sorted - Peck's Hebeloma revisited.

Charles Horton Peck described some 2700 species of North American fungi in the 19th and early 20th centuries. Among these were 31 species that he described as Hebeloma or that later authors recombined into Hebeloma. These 31 taxa have been analyzed morphologically and molecularly, as far as possible. For six of these species, lectotypes are designated. For twelve species, ITS sequences (some partial) were generated. Thirteen of the species analyzed are Hebeloma, as the genus is delimited today. Of these 13, nine are regarded as 'current', i.e. are names that should be accepted and used. Of the remaining four, three are synonymized with earlier Peck species and one with the generic type H. mesophaeum. Numerous Hebeloma species described from America are synonymized with some of Peck's species, such as H. albidulum, H. album, H. colvinii, H. excedens, H. palustre, H. sordidulum, and H. velatum; Peck's H. album, H. palustre, and H. velatum are earlier names for H. fragilipes, H. clavulipes, and H. dunense, respectively. All three names were in current use and described from Europe. The 18 species that are not Hebeloma belong to a range of genera: Agrocybe, Hemistropharia, Inocybe, Inosperma, Naucoria, and Pholiota; three species that were not previously recombined into their respective genera are here recombined and one species, Hebeloma commune is synonymized with Pholiota lenta. Two taxa, that are not Hebeloma, remain unresolved. Sixty later Hebeloma taxa described from North America are revised and synonymized with Peck species and seven with H. mesophaeum, 36 of these supported by ITS (some partial) sequence data. Updates on two species, H. petrakii and H. remyi, from Europe, are also given, and a lectotype and epitype selected for the latter.

Analysis of N-glycosylation in fungal l-amino acid oxidases expressed in the methylotrophic yeast Pichia pastoris.

l-amino acid oxidases (LAAOs) catalyze the oxidative deamination of l-amino acids to corresponding α-keto acids. Here, we describe the heterologous expression of four fungal LAAOs in Pichia pastoris. cgLAAO1 from Colletotrichum gloeosporioides and ncLAAO1 from Neurospora crassa were able to convert substrates not recognized by recombinant 9His-hcLAAO4 from the fungus Hebeloma cylindrosporum described earlier thereby broadening the substrate spectrum for potential applications. 9His-frLAAO1 from Fibroporia radiculosa and 9His-laLAAO2 from Laccaria amethystine were obtained only in low amounts. All four enzymes were N-glycosylated. We generated mutants of 9His-hcLAAO4 lacking N-glycosylation sites to further understand the effects of N-glycosylation. All four predicted N-glycosylation sites were glycosylated in 9His-hcLAAO4 expressed in P. pastoris. Enzymatic activity was similar for fully glycosylated 9His-hcLAAO4 and variants without one or all N-glycosylation sites after acid activation of all samples. However, activity without acid treatment was low in a variant without N-glycans. This was caused by the absence of a hypermannosylated N-glycan on asparagine residue N54. The lack of one or all of the other N-glycans was without effect. Our results demonstrate that adoption of a more active conformation requires a specific N-glycosylation during biosynthesis.

Benefits provided by four ectomycorrhizal fungi to Pinus taeda under different external potassium availabilities.

Ectomycorrhizal fungi contribute to the nutrition of many woody plants, including those in the Pinaceae family. Loblolly pine (Pinus taeda L.), a native species of the Southeastern USA, can be colonized by multiple species of ectomycorrhizal fungi. The role of these symbionts in P. taeda potassium (K+) nutrition has not been previously investigated. Here, we assessed the contribution of four ectomycorrhizal fungi, Hebeloma cylindrosporum, Paxillus ammoniavirescens, Laccaria bicolor, and Suillus cothurnatus, in P. taeda K+ acquisition under different external K+ availabilities. Using a custom-made two-compartment system, P. taeda seedlings were inoculated with one of the four fungi, or kept non-colonized, and grown under K+-limited or -sufficient conditions for 8 weeks. Only the fungi had access to separate compartments in which rubidium, an analog tracer for K+, was supplied before harvest. Resulting effects of the fungi were recorded, including root colonization, biomass, and nutrient concentrations. We also analyzed the fungal performance in axenic conditions under varying supply of K+ and sodium. Our study revealed that these four ectomycorrhizal fungi are differentially affected by external K+ and sodium variations, that they are not able to provide similar benefits to the host P. taeda in our growing conditions, and that rubidium may be used with some limitations to estimate K+ transport from ectomycorrhizal fungi to colonized plants.

Fungal Shaker-like channels beyond cellular K+ homeostasis: A role in ectomycorrhizal symbiosis between Hebeloma cylindrosporum and Pinus pinaster.

Potassium (K+) acquisition, translocation and cellular homeostasis are mediated by various membrane transport systems in all organisms. We identified and described an ion channel in the ectomycorrhizal fungus Hebeloma cylindrosporum (HcSKC) that harbors features of animal voltage-dependent Shaker-like K+ channels, and investigated its role in both free-living hyphae and symbiotic conditions. RNAi lines affected in the expression of HcSKC were produced and used for in vitro mycorrhizal assays with the maritime pine as host plant, under standard or low K+ conditions. The adaptation of H. cylindrosporum to the downregulation of HcSKC was analyzed by qRT-PCR analyses for other K+-related transport proteins: the transporters HcTrk1, HcTrk2, and HcHAK, and the ion channels HcTOK1, HcTOK2.1, and HcTOK2.2. Downregulated HcSKC transformants displayed greater K+ contents at standard K+ only. In such conditions, plants inoculated with these transgenic lines were impaired in K+ nutrition. Taken together, these results support the hypothesis that the reduced expression of HcSKC modifies the pool of fungal K+ available for the plant and/or affects its symbiotic transfer to the roots. Our study reveals that the maintenance of K+ transport in H. cylindrosporum, through the regulation of HcSKC expression, is required for the K+ nutrition of the host plant.

Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation.

l-amino acid oxidases (LAAOs) are flavoenzymes that catalyze the oxidative deamination of l-amino acids to the corresponding α-keto acids, ammonia, and hydrogen peroxide. Here, we show the overexpression, purification, and the characterization of LAAO4 from the fungus Hebeloma cylindrosporum in the yeast Pichia pastoris with a 9His-tag and compare this with the recently characterized 6His-hcLAAO4 expressed in E. coli. The expression of the enzyme with an ER-signal sequence in P. pastoris resulted in a glycosylated, secreted protein. The enzymatic activity without activation was higher after expression in P. pastoris compared to E. coli. Due to treatment with acidic pH, a striking increase of activity could be detected for both expression systems resulting in similar specific activities after acid activation. Regarding the substrate spectrum, temperature stability, Km, and vmax values, hcLAAO4 showed very few differences when produced in these two expression systems. A higher yield of hcLAAO4 could be obtained by fermentation.

New insights into HcPTR2A and HcPTR2B, two high-affinity peptide transporters from the ectomycorrhizal model fungus Hebeloma cylindrosporum.

While plants mainly rely on the use of inorganic nitrogen sources like ammonium and nitrate, soil-borne microorganisms like the ectomycorrhizal fungus Hebeloma cylindrosporum can also take up soil organic N in the form of amino acids and peptides that they use as nitrogen and carbon sources. Following the previous identification and functional expression in yeast of two PTR-like peptide transporters, the present study details the functions and substrates of HcPTR2A and HcPTR2B by analysing their transport kinetics in Xenopus laevis oocytes. While both transporters mediated high-affinity di- and tripeptide transport, HcPTR2A also showed low-affinity transport of several amino acids-mostly hydrophobic ones with large side chains.

Process properties of an l-amino acid oxidase from Hebeloma cylindrosporum for the synthesis of phenylpyruvic acid from l-phenylalanine.

The biocatalytic oxidation of amino acids represents an attractive approach towards the synthesis of α-keto acids, which are interest for various industrial applications. As l-amino acids are readily available from fermentation processes, these natural amino acids can serve as substrates in combination with an l-amino acid oxidase. Besides an aqueous phase as reaction medium, a further advantage of such a process is the utilization of air as oxidation agent. In this study, we studied the organic-synthetic properties of a literature-known recombinant l-amino acid oxidase from the fungus Hebeloma cylindrosporum with respect to its suitability to catalyze the formation of α-keto acids exemplified for the synthesis of phenylpyruvic acid starting from l-phenylalanine as a substrate. In our study the enzyme displayed a reasonable operational stability in the reaction system and as well as promising applicability data with respect to substrate and product inhibition. In a biotransformation, 20 mM of substrate were converted after 4 h reaction. The formation of undesired by-products was suppressed using a commercially available catalase enzyme.

Adventurous cuisine in Laos: Hebeloma parvisporum, a new species in Hebeloma section Porphyrospora.

Hebeloma parvisporum is described as new and placed within H. sect. Porphyrospora. This mushroom is sold as an edible in markets of Laos under the local name "wai khom." Hebeloma sect. Porphyrospora is discussed and expanded to include the species formerly included in the genus Anamika and recently transferred to Hebeloma. Hebeloma sect. Porphyrospora currently comprises 16 species, 14 of which are known only from the western Pacific and Indian subcontinent. All species in this section share the character of having red-brown spores when fresh, atypical for other sections of Hebeloma, which causes the lamellae to be red-brown. However, this red-brown color fades when the material is dried. The close links, morphologically and molecularly, between H. parvisporum and other members of H. sect. Porphyrospora, particularly H. victoriense, are shown.

Arsenic toxicity and its mitigation in ectomycorrhizal fungus Hebeloma cylindrosporum through glutathione biosynthesis.

Arsenic (As) contamination is one of the most daunting environmental problem bothering the whole world. Exploring a suitable bioremediation technique is an urgent need of the hour. The present study focusses on scrutinizing the ectomycorrhizal (ECM) fungus for its potential role in As detoxification and understanding the molecular mechanisms responsible for its tolerance. When exposed to increasing concentrations of external As, the ECM fungus H. cylindrosporum accumulated the metalloid intracellularly, inducing the glutathione biosynthesis pathway. The genes coding for GSH biosynthesis enzymes, γ-glutamylcysteine synthetase (Hcγ-GCS) and glutathione synthetase (HcGS) were highly regulated by As stress. Arsenic coordinately upregulated the expression of both Hcγ-GCS and HcGS genes, thus resulting in increased Hcγ-GCS and HcGS protein expressions and enzyme activities, with substantial increase in intracellular GSH. Functional complementation of the two genes (Hcγ-GCS and HcGS) in their respective yeast mutants (gsh1Δ and gsh2Δ) further validated the role of both enzymes in mitigating As toxicity. These findings clearly highlight the potential importance of GSH antioxidant defense system in regulating the As induced responses and its detoxification in ECM fungus H. cylindrosporum.

Heterologous expression of Zn-binding peptide RaZBP1 from Russula bresadolae does not overcome Zn and Cd detoxification mechanisms in Hebeloma mesophaeum.

Homeostatic mechanisms preventing the toxicity of heavy metal ions in cells involve, among others, compartmentalization and binding with peptidaceous ligands, particularly the cysteinyl-rich metallothioneins (MTs). We have previously shown that in natural conditions Zn-overaccumulating ectomycorrhizal (EM) fungus Russula bresadolae stores nearly 40% of Zn bound with cysteinyl- and hystidyl-containing RaZBP peptides, which resemble MTs, while the detoxification of Zn and Cd in EM Hebeloma mesophaeum relies upon compartmentalization in small vesicles and vacuoles, respectively. Here, we examined the performance of RaZBP1 gene expressed in H. mesophaeum mycelium with respect to handling of Zn and Cd. Expression of RaZBP1 impaired growth of the mycelium on low-Zn medium by 60%, the growth was partly ameliorated upon the addition of Zn and remained considerable up to 2 mmol/L Zn, while the growth of the wild-type and control mycelia transformed with empty T-DNA was severely reduced in the presence of 0.5 mmol/L Zn; furthermore, RaZBP1 slightly added to Cd tolerance in the range of Cd concentrations of 0.625 to 8 µmol/L. Staining of Zn- or Cd-exposed hyphal cells with Zn- or Cd-specific fluorescent tracers did not indicate that the expression of RaZBP1 would redirect the flow of the metals away from their innate sinks. Size exclusion chromatography of extracted metal species revealed that the complexes corresponding to Zn/Cd-RaZBP1 are present only in minute levels. Considering that RaZBP1 inhibited growth at low Zn, and despite the benefit that it provided to H. mesophaeum in the presence of high Zn and moderate Cd, these data indicate that the binding of excess Zn and Cd with RaZBP1 is not a trait that would be outright transmitted to H. mesophaeum.

A Cr(VI)-tolerant strain, Pisolithus sp1, with a high accumulation capacity of Cr in mycelium and highly efficient assisting Pinus thunbergii for phytoremediation.

Ectomycorrhizal (ECM) fungi can improve the growth of pine trees and enhance their tolerance to heavy metal stress, and may also be useful during the afforestation and phytoremediation of polluted regions with pine trees. Hebeloma vinosophyllum (Cr(VI)-sensitive strain) and Pisolithus sp1 ((Cr(VI)-tolerant strain) were selected through liquid culture experiment, and were used in symbiosis with Japanese black pine (Pinus thunbergii) in pot experiments, to determine their potential for improving phytoremediation of Cr(VI)-contaminated soils. Our results indicated that Pisolithus sp1 also had a significantly higher accumulation of Cr than H. vinosophyllum in mycelium under the same Cr(VI) treatments in liquid culture experiment. The tolerance index of Pisolithus sp1 ECM seedlings' shoots and roots to Cr(VI) were significantly higher than that of H. vinosophyllum ECM and non-ectomycorrhizal (NM) seedlings while the total accumulated Cr per seedling in Pisolithus sp1 ECM seedlings were 1.50-1.96 and 2.83-27.75 fold higher that of H. vinosophyllum ECM and NM seedlings, respectively, within 0-800 mg kg-1 Cr(VI) treatments in pot experiments. In addition, the significant differences ratios of photosynthetic rate, stomatal conductance, transpiration rate and intercellular CO2 concentration between Pisolithus sp1 ECM and NM seedlings were significantly higher than those between H. vinosophyllum ECM and NM seedlings under 400 and 800 mg kg-1 Cr(VI) treatments. Compared with the control (no plant), and planting NM or H. vinosophyllum ECM seedlings, the planting of Pisolithus sp1 ECM seedlings significantly reduced the percentage content of exchangeable Cr in the soil.

Different cadmium tolerance of two isolates of Hebeloma mesophaeum showing different basal expression levels of metallothionein (HmMT3) gene.

Hebeloma mesophaeum is an ectomycorrhizal fungus frequently associated with metal disturbed environments. In this work, we examined Ag, Cd, and Zn tolerance of H. mesophaeum isolates from heavy metal-polluted (isolate Prib) and clean (isolate Rez) sites. Both mycelia showed essentially the same level of Ag and Zn tolerance, but Prib was more Cd tolerant. In short-term exposures, Prib accumulated slightly less Cd than Rez. Size exclusion chromatography of cell-free extracts and fluorescence microscopy of hyphae with a Cd-specific fluorescent tracer revealed that substantial proportion of Cd was contained in the vacuoles in both isolates. Considering that the proportion of Cd associated with fractions attributable to Cd complexes with cytosolic, metallothionein (MT) peptides was higher in Prib, we examined the copy number and basal levels of HmMTs genes in Rez and Prib. While no difference between the isolates was observed in the gene copy numbers and basal levels of HmMT1 transcripts, the basal transcription of HmMT3 was 3-fold higher in Prib. These observations suggest that MTs provide in Prib better protection against Cd. Furthermore, the higher Cd tolerance in Prib can be to some extent also supported by the efflux or reduced uptake of Cd in the hyphae.

Expression, characterization, and site-specific covalent immobilization of an L-amino acid oxidase from the fungus Hebeloma cylindrosporum.

L-Amino acid oxidases (LAAOs) are flavoproteins, which use oxygen to deaminate L-amino acids and produce the corresponding α-keto acids, ammonia, and hydrogen peroxide. Here we describe the heterologous expression of LAAO4 from the fungus Hebeloma cylindrosporum without signal sequence as fusion protein with a 6His tag in Escherichia coli and its purification. 6His-hcLAAO4 could be activated by exposure to acidic pH, the detergent sodium dodecyl sulfate, or freezing. The enzyme converted 14 proteinogenic L-amino acids with L-glutamine, L-leucine, L-methionine, L-phenylalanine, L-tyrosine, and L-lysine being the best substrates. Methyl esters of these L-amino acids were also accepted. Even ethyl esters were converted but with lower activity. Km values were below 1 mM and vmax values between 19 and 39 U mg-1 for the best substrates with the acid-activated enzyme. The information for an N-terminal aldehyde tag was added to the coding sequence. Co-expressed formylglycine-generating enzyme was used to convert a cysteine residue in the aldehyde tag to a Cα-formylglycine residue. The aldehyde tag did not change the properties of the enzyme. Purified Ald-6His-hcLAAO4 was covalently bound to a hexylamine resin via the Cα-formylglycine residue. The immobilized enzyme could be reused repeatedly to generate phenylpyruvate from L-phenylalanine with a total turnover number of 17,600 and was stable for over 40 days at 25 °C.

Cadmium induced glutathione bioaccumulation mediated by γ-glutamylcysteine synthetase in ectomycorrhizal fungus Hebeloma cylindrosporum.

Ectomycorrhizal fungi hold a potential role in bioremediation of heavy metal polluted areas because of its metal accumulation and detoxification property. We investigated the cadmium (Cd) induced bioaccumulation of glutathione (GSH) mediated by γ-glutamylcysteine synthetase (γ-GCS) in the ectomycorrhizal fungus Hebeloma cylindrosporum. In H. cylindrosporum, a demand driven synthesis of GSH has been observed in response to Cd. The expression and enzyme activity of H. cylindrosporum γ-GCS (Hcγ-GCS) increased as a function of external Cd stress resulting in increased GSH production. The function of Hcγ-GCS in providing heavy metal tolerance to H. cylindrosporum was justified by complementing the gene in gsh1Δ mutant of Saccharomyces cerevisiae. The metal sensitive mutant gsh1Δ successfully restored its metal tolerance ability when transformed with Hcγ-GCS gene. Sequence analysis of Hcγ-GCS showed homology with most of the reported γ-GCS proteins from basidiomycetes family. The active site of the Hcγ-GCS protein is composed of amino acids that were found to be conserved not only in fungi, but also in plants and mammals. From these results, it was concluded that Hcγ-GCS plays an important role in bioaccumulation of GSH, which is a core component in the mycorrhizal defense system under Cd stress for Cd homeostasis and detoxification.

Whole-plant frost hardiness of mycorrhizal (Hebeloma sp. or Suillus luteus) and non-mycorrhizal Scots pine seedlings.

Ectomycorrhizal trees are common in the cold regions of the world, yet the role of the mycorrhizal symbiosis in plant cold tolerance is poorly known. Moreover, the standard methods for testing plant frost hardiness may not be adequate for roots and mycorrhizas. The aims of this study were to compare the frost hardiness of mycorrhizal and non-mycorrhizal Scots pine (Pinus sylvestris L.) seedlings and to test the use of reverse-flow root hydraulic conductance (Kr) measurement for root frost hardiness determination. Mycorrhizal (Hebeloma sp. or Suillus luteus) and non-mycorrhizal seedlings were grown in controlled-environment chambers for 13 weeks. After this, half of the plants were allotted to a non-hardening treatment (long day and high temperature, same as during the preceding growing season) and the other half to a hardening (short day and low temperature) 'autumn' treatment for 4 weeks. The intact seedlings were exposed to whole-plant freezing tests and the needle frost hardiness was measured by relative electrolyte leakage (REL) method. The seedlings were grown for three more weeks for visual damage assessment and Kr measurements using a high-pressure flow meter (HPFM). Mycorrhizas did not affect the frost hardiness of seedlings in either hardening treatment. The effect of the hardening treatment on frost hardiness was shown by REL and visual assessment of the aboveground parts as well as Kr of roots. Non-mycorrhizal plants were larger than mycorrhizal ones while nitrogen and phosphorus contents (per unit dry mass) were similar in all mycorrhiza treatments. In plants with no frost exposure, the non-mycorrhizal treatment had higher Kr. There was no mycorrhizal effect on plant frost hardiness when nutritional effects were excluded. Further studies are needed on the role of mycorrhizas especially in the recovery of growth and nutrient uptake in cold soils in the spring. The HPFM is useful novel method for assessment of root damage.

HcPT1.2 participates in Pi acquisition in Hebeloma cylindrosporum external hyphae of ectomycorrhizas under high and low phosphate conditions.

Ectomycorrhizal fungi improve tree phosphorus nutrition through transporters specifically localized at soil-hyphae and symbiotic interfaces. In the model symbiosis between the fungus Hebeloma cylindrosporum and the maritime pine (Pinus pinaster), several transporters possibly involved in phosphate fluxes were identified, including three H+:Pi transporters. Among these three, we recently unraveled the function of one of them, named HcPT2, in both pure culture and symbiotic interaction with P. pinaster. Here we investigated the transporter named HcPT1.2, by analyzing inorganic phosphate transport ability in a yeast complementation assay, assessing its expression in the fungus associated or not with the plant, and immunolocalizing the proteins in ectomycorrhizas. We also evaluated the effect of external Pi concentration on expression and localization of HcPT1.2. Our results revealed that HcPT1.2 is involved in Pi acquisition by H. cylindrosporum mycelium, irrespective of the external Pi concentrations.

HcTOK1 participates in the maintenance of K+ homeostasis in the ectomycorrhizal fungus Hebeloma cylindrosporum, which is essential for the symbiotic K+ nutrition of Pinus pinaster.

Most land plants rely on root symbioses to complement or improve their mineral nutrition. Recent researches have put forward that mycorrhizal fungi efficiently absorb and transfer potassium (K+) from the soil to host plant roots, but the molecular mechanisms involved are not completely elucidated yet. We have recently revealed that K+ is likely released from the fungal Hartig net to the plant by TOK channels in the ectomycorrhizal model Hebeloma cylindrosporum - Pinus pinaster. H. cylindrosporum harbours three TOK members. Herein, we report that one of them, HcTOK1, has similar features than the yeast ScTOK1. Moreover, we propose a role for this channel in the transport of K+ from the medium to ectomycorrhizal roots under K+ starvation.