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PubMed1999

IgE-binding proliferative responses and skin test reactivity to Cop c 1, the first recombinant allergen from the basidiomycete Coprinus comatus.

BACKGROUND: Basidiomycetes spores are ubiquitously distributed, found throughout the year in outdoor and indoor air, and represent relevant sources of aeroallergens associated with allergy and asthma. OBJECTIVE: Cloning and characterization of Coprinus comatus (shaggy cap mushroom) allergens is essential to elucidate their molecular characteristics and to improve the diagnosis of allergy. METHODS: A complementary DNA (cDNA) library of C comatus displayed on phage surface was screened with sera of basidiomycete-sensitized individuals. Subcloning and high-level expression of one of the enriched cDNAs allowed the isolation of a [His](6)-tagged recombinant protein formally termed rCop c 1. The allergenic properties of rCop c 1 were investigated in vitro by ELISA, inhibition experiments, immunoblots, and proliferation assays and in vivo by skin tests. RESULTS: The rCop c 1-encoding cDNA spans 435 bp and contains an open reading frame of 246 bp, predicting a protein of 8.96 kd without significant sequence homology to known proteins. Immunoblots with [His](6)-rCop c 1 fusion protein show a background free IgE-binding band of the expected size that can be completely inhibited by crude C comatus extracts in ELISA. rCop c 1 induced specific proliferative responses in PBMCs of C comatus-sensitized individuals. The incidence of sensitization to rCop c 1 among 92 sera of basidiomycete-sensitized individuals tested in ELISA was 25%, indicating that Cop c 1 is an intermediate allergen. However, prick tests showed that less than 2 pmol of the rCop c 1 protein was able to induce strong specific skin reactions in sensitized individuals. CONCLUSIONS: rCop c 1, the first cloned allergen from the genus Coprinus, fulfills all the criteria required to be classified as a clinically relevant allergen. The data demonstrate at the molecular level the presence of sensitizing molecules among Basidiomycetes, the most important source contributing to the total spore load in the outdoor air.

Allergens↗

PubMed2005

Screening of basidiomycetes and xylariaceous fungi for lignin peroxidase and laccase gene-specific sequences.

Lignin peroxidase and laccase gene-specific PCR primers were used to screen 38 diverse basidiomycetes and xylariaceous fungi. Lignin peroxidase gene-specific sequences were obtained for basidiomycetes only and were highly divergent. Possession of laccase genes was relatively widespread among basidiomycetes, and is shown for the first time in Xylariaceae. All sequences were highly conserved with no variation resulting in changes to predicted amino acid sequence. Those basidiomycetes shown to possess lignin peroxidase and laccase genes also produced the enzyme in vitro. Conversely none of the xylariaceous fungi shown to possess laccase genes were able to do so, whilst others decolorized Poly R yet yielded no PCR amplicons.

Amino Acid Sequence↗

PubMed2005

Molecular data confirm that Omphalina foliacea is a lichen-forming basidiomycete.

We examined the phylogenetic position of Omphalina foliacea and its putative closest relative, the lichen-forming basidiomycete Lichenomphalia lobata. Both species are restricted to specific habitats in the high andine regions of tropical America. We generated nuclear ITS and LSU rDNA sequences of two collections of each species and analysed the data together with sequences of omphalinoid basidiomycetes retrieved from GenBank using a Bayesian and a maximum parsimony approach. Our analyses show that both, O. foliacea and L. lobata are lichenized basidiomycetes. L. lohata is related to other species of the genus Lichenomphalia, whereas O. foliacea is not closely related to other Omphalina or Lichenomphalia species, but probably belongs in the hymenochaetoid clade. The analysis of a nuclear LSU data set of a broader sampling of hymenochaetoid basidiomycetes supported the placement of O. foliacea in this clade, but did not reveal any close relative.

Basidiomycota↗

PubMed1993

ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.

We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.

Ascomycota↗

PubMed1996

Isolation of saprophytic basidiomycetes from soil.

A method with the combined advantages of soil particle washing, selective inhibitors, and an indicator substrate was developed to isolate saprophytic basidiomycetes from soil. Organic particles were washed from soil and plated on a medium containing lignin, guaiacol, and benomyl, which reduced mold growth and allowed detection of basidiomycetes producing laccase or peroxidase. The 64 soil samples yielded 67 basidiomycete isolates, representing 51 groups on the basis of morphology and physiology. This method should facilitate investigations into the biodiversity of soil basidiomycetes and yield organisms that are useful in bioremediation of soils contaminated with pesticides or other recalcitrant aromatic compounds.

Journal Article↗

PubMed2006

Necrotrophic mycoparasitism of Botrytis cinerea by cellulolytic and ligninocellulolytic Basidiomycetes.

Twenty-six isolates representing 17 species of aphyllophoraceous, wood-decaying Basidiomycetes and five species of agaricoid, turf-borne, thatch-decaying Basidiomycetes were screened for their abilities to degrade cellulose, lignin, and melanin by using colorimetric degradation assays on agar media. Selected ligninocellulolytic Basidiomycetes capable of degrading melanin were screened for antagonism of Botrytis cinerea Per.:Fr. The greatest inhibition of Botrytis colony and hyphal growth in vitro was observed in confrontations with Irpex lacteus (Fr.) Fr., Trametes versicolor (L.:Fr.) Pilat, and Chondrostereum purpureum (Pers.:Fr.) Pouzar. Hyphal interference and necrotrophic mycoparasitism by these ligninocellulolytic Basidiomycetes were recognized microscopically as coagulation and degeneration of Botrytis cytoplasm and as coiling and invasion of hyphae, conidiophores, and conidia, respectively. Sclerotia of B. cinerea were killed and parasitized in agar media, straw mulch, or moist sand infested separately with these three mycoparasites.

Basidiomycota↗

PubMed1996

Screening of selected basidiomycetes for inhibitory activity on neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE).

Aqueous extracts of 27 basidiomycetes were investigated for their ability to inhibit the activity of angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). The extracts of 5 fungi inhibited both, ACE and NEP activity, another 18 extracts showed inhibition of the NEP activity whereas only 1 basidiomycete inhibited the ACE activity exclusively. The IC50 values for the ACE inhibition are rather high (between 200 and 1500 micrograms/ml) in comparison to the IC50 of the NEP inhibition (between 40 and 2000 micrograms/ml). These results indicate that the basidiomycetes investigated seem to have a higher potential for the inhibition of the activity of NEP than of ACE. In general, basidiomycetes are a new source for inhibitors of metalloendopeptidases. Resulting from the isolation and characterization of these compounds new leading structures are expectable.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1999

The lysine-specific proteinase from Armillaria mellea is a member of a novel class of metalloendopeptidases located in Basidiomycetes.

The fruiting body of the basidiomycete fungus Armillaria mellea produces a lysine-specific proteinase which exhibits both potent fibrinolytic activity and a remarkable resistance to denaturing agents. An improved purification protocol has been developed for this enzyme and the sequence of the 26 N-terminal amino acid residues of the pure protein has been determined by gas-phase sequencing. Searches of the SwissProt database showed that the N-terminal sequence of A. mellea proteinase is highly similar to those of lysine-specific metalloendopeptidases from the basidiomycetes Grifola frondosa and Pleurotus ostreatus. These results support the view that the A. mellea proteinase is a member of a novel class of lysine-specific metalloendopeptidases which may be exclusive to basidiomycete fungi.

Amino Acid Sequence↗

PubMed1992

Sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase genes from the basidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus.

GPD genes encoding glyceraldehyde-3-phosphate dehydrogenase were isolated from the homobasidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus. All three species contain one transcriptionally active GPD gene, but A. bisporus also contains an inactive GPD gene (tandemly linked to the active gene). These genes contain 5-9 introns located at conserved positions, differing (except in one case) from intron positions in ascomycetous GPD genes. The predicted amino-acid sequences of the proteins encoded by the three active GPD genes are highly homologous. A comparison with protein sequences from filamentous ascomycetes shows a clear distinction, whereas the GPD genes from ascomycetous yeasts are quite distinct from both the filamentous ascomycetes and basidiomycetes. Promoter regions of ascomycetous GPD genes do not correspond to those of the GPD genes of basidiomycetes which may (partly) explain poor expression in basidiomycetes of introduced genes driven by an ascomycete GPD promoter.

Amino Acid Sequence↗

PubMed2003

Identification of Ty1-copia retrotransposons in three ectomycorrhizal basidiomycetes: evolutionary relationships and use as molecular markers.

We amplified by PCR and sequenced 46 partial Ty1- copia reverse transcriptase (RT) sequences from the ectomycorrhizal basidiomycetes Pisolithus and Laccaria bicolor and the host tree Eucalyptus globulus. Phylogenetic analyses indicated that these sequences represent a new class of Ty1- copia RT, characteristic of basidiomycetes but related to plant Ty1- copia retrotransposons. To generate fingerprints of L. bicolor strains, outward facing PCR primers annealing to RTs were designed. This method, which is a modification of the inter-retrotransposon amplified polymorphism (IRAP) analysis, enables the detection of polymorphisms or changes within the insertion sites of Ty1- copia elements in the genome. Using this method, we investigated whether the transposition of Ty1- copia elements was related to the somaclonal variation observed in L. bicolor S238, an inoculant strain used in French Douglas-fir plantations. Data indicated that no differences in the IRAP fingerprints were detected in phenotypic variants of L. bicolor S238. We reported here for the first time the presence of Ty1- copia retrotransposon sequences in basidiomycetes, which resulted in suitable targets for developing new molecular markers.

Amino Acid Sequence↗

PubMed2005

Production of volatile organic sulfur compounds (VOSCs) by basidiomycetous yeasts.

Thirty-seven basidiomycetous yeasts belonging to 30 species of seven genera were grown on media containing l-cysteine or l-methionine as sole nitrogen sources with the objective of evaluating volatile organic sulfur compound (VOSC) production. The headspace of yeast cultures was analyzed by the solid-phase microextraction (SPME) sampling method, and volatile compounds were quantified and identified by GC-MS techniques. Ten strains assimilating L-methionine produced the following VOSCs: 3-(methylthio)-1-propanol, methanethiol, S-methyl thioacetate, dimethyl disulfide, dimethyl trisulfide, allyl methyl sulphide and 4,5-dihydro-3(2H)-thiophenone. Production was <1 mgl(-1) except for 3-(methylthio)-1-propanol of which between 40 and 400 mgl(-1) was synthesized. Higher alcohols (isobutyl alcohol, isoamyl alcohol and active amyl alcohol) and esters (ethyl acetate, ethyl propionate, n-propyl acetate, isobutyl acetate, n-propyl propionate, n-butyl acetate, isoamyl acetate, amyl acetate, isoamyl propionate, amyl propionate and 2-phenylmethyl acetate) were also sporadically produced. This is the first report of VOSCs production by basidiomycetous yeasts. Consequently, basidiomycetous yeasts may be considered an interesting new group of microbial VOSCs producers for the flavor industry.

Basidiomycota↗

PubMed2002

Endopolygalacturonase is encoded by a multigene family in the basidiomycete Chondrostereum purpureum.

The basidiomycete Chondrostereum purpureum produces several plant cell wall-degrading enzymes, including endopolygalacturonase (endoPG). Degenerate oligonucleotide primers were designed according to conserved regions of endoPG genes from various fungi, plants, and bacteria and used to amplify members of this gene family from C. purpureum. Four different amplification products showed significant similarity to known endoPGs and were used as hybridization probes to screen a library of genomic DNA sequences and to retrieve five full-length endoPG genes (epgA, epgB1, epgB2, epgC, and epgD). The identities between the deduced polypeptides for epgA, epgB1, epgC, and epgD ranged from 61.8 to 80.0%, while the deduced polypeptides for epgB1 and epgB2 shared 97.1% identity. Phylogenetic analysis suggested that the duplication of existing endoPG genes occurred after the divergence of the ascomycetes and basidiomycetes. C. purpureum is the first basidiomycete fungus for which the endoPG gene family has been described.

Agaricales↗

PubMed2005

Basidiomycete mycelia in forest soils: dimensions, dynamics and roles in nutrient distribution.

Basidiomycete mycelia are ubiquitous in forest soils where they fulfil a range of key ecological functions. Population studies, based largely on basidiome collections, indicate that mycelia of many ectomycorrhizal and saprotrophic basidiomycetes can spread vegetatively for considerable distances through soil, but the extent to which these become physically or physiologically fragmented is unclear. This review considers aspects of the distribution, dynamics and translocatory activities of individual basidiomycete mycelia in forest soil, highlighting current gaps in our understanding and possible ways to address these.

Basidiomycota↗

PubMed2000

Screening of basidiomycetes for antimicrobial activities.

As a part of a screening programme developed to evaluate the antimicrobial activity of basidiomycetes, 317 isolates representing 204 species collected in Spain were screened against a range of human clinical pathogens and laboratory controls. Extracts from 45% of the isolates, representing 109 species, showed antimicrobial activity. Antibacterial activity was more pronounced than antifungal activity. The proportion of extracts from basidiomycetes showing antimicrobial activity was similar to or above that obtained for representative orders of Ascomycetes, such as Pezizales and Xylariales, but lower than that produced by members of the orders Diaporthales, Eurotiales, Hypocreales, Leotiales and Sordariales. Suprageneric taxa (orders and families) did not show pronounced differences in their antimicrobial activities though such differences were observed at the genus level, suggesting that the ability to produce these bioactive compounds is not homogenously distributed amongst the basidiomycetes. Isolates from some species showed large differences in their ability to produce metabolites with antimicrobial activity, possibly reflecting genetic differences at the infraspecific level.

Anti-Bacterial Agents↗

PubMed2001

Molecular phylogenetics of the genus Rhodotorula and related basidiomycetous yeasts inferred from the mitochondrial cytochrome b gene.

Phylogenetic relationships of basidiomycetous yeasts, especially of the genus Rhodotorula, were studied using partial sequences of the mitochondrial cytochrome b gene. The results demonstrated that the basidiomycetous yeasts under investigation distributed into two main clusters: one containing Tremellales, Filobasidiales and their anamorphs and the other containing Ustilaginales, Sporidiales and their anamorphs. This clustering in turn correlates with cell wall biochemistry, presence or absence of xylose, and septal ultrastructure, dolipore or simple pore. Bullera, Bulleromyces, Filobasidiella, Cryptococcus and Trichosporon, yeasts of the former cluster, contain xylose in the cell wall and have dolipore septa. In contrast yeasts of the latter cluster, which included Bensingtonia, Erythrobasidium, Leucosporidium, Malassezia, Rhodosporidium, Rhodotorula, Sporidiobolus, Sporobolomyces and Ustilago, have no xylose in the cell wall and have a simple pore septum. Yeasts of the latter group could be further divided into four clades (A-D). Species of Rhodotorula were distributed in all of these clades, indicating the polyphyletic nature of the genus. A limited number of Rhodotorula species demonstrated identical sequences, for example Rhodotorula bacarum and Rhodotorula foliorum, Rhodotorula fujisanensis and Rhodotorula futronensis, Rhodotorula glutinis var. dairenensis and Rhodotorula mucilaginosa. However, all the other test species of the genus Rhodotorula were well separated based on their 396 bp nucleotide sequences. These results demonstrate the effectiveness of the use of cytochrome b sequences for both species identification and the study of phylogenetic relationships among basidiomycetous yeasts.

Amino Acid Sequence↗

PubMed1994

Prevalence of basidiomycete allergy in the USA and Europe and its relationship to allergic respiratory symptoms.

A total of 701 adults living in the USA or Western Europe who had symptoms of allergic respiratory disease were skin prick tested with extracts prepared from eight basidiomycetes species and four Fungi Imperfecti species. In these subjects, the presence of asthma, rhinitis, or both was determined by questionnaire. Overall, 178/701 (25.4%) of the participants reacted to at least one basidiomycete extract. There was no difference in the prevalence of reactivity detected in the USA and Europe (P < 0.005); however, the prevalence of reactors in individual centers from both areas varied significantly. Psilocybe cubensis was the most potent allergen source in both the USA (12.3% reacted) and Europe (16.0%). Pleurotus ostreatus was second overall (10.6%) and in the USA (10.7%), and third in Europe (10.3%). Pisolithus tinctorius and Coprinus quadrifidus produced the least potent allergens, with only 5.4% of the population reacting. There was a significant relationship (P < 0.005) between basidiospore reactivity and the presence of atopy, asthma, and asthma and rhinitis. Basidiospore reactivity was not associated with the presence of rhinitis alone (P = 0.312). These results suggest that basidiomycetes are important sources of aeroallergens in geographically disparate regions and may be particularly important in patients with asthma.

Adult↗

PubMed1996

The molecular basis for the natural resistance of the cytochrome bc1 complex from strobilurin-producing basidiomycetes to center Qp inhibitors.

Mitochondria from the strobilurin A producing basidiomycetes Strobilurus tenacellus and Mycena galopoda exhibit natural resistance to (E)-beta-methoxyacrylate inhibitors of the ubiquinol oxidation center(center Qp) of the cytochrome bc1 complex. Isolated cytochrome bc1 complex from S. tenacellus was found to be highly similar to that of Saccharomyces cerevisiae with respect to subunit composition, as well as spectral characteristics and midpoint potentials of the heme centers. To understand the molecular basis of natural resistance, we determined the exon/intron organization and deduced the sequences of cytochromes b from S. tenacellus, M. galopoda and a third basidiomycete, Mycena viridimarginata, which produces no strobilurin A. Comparative sequence analysis of two regions of cytochrome b known to contribute to the formation of center Qp suggested that the generally lower sensitivity of all three basidiomycetes was due to the replacement of a small amino acid residue in position 127 by isoleucine. For M. galopoda replacement of Gly143 by alanine and Gly153 by serine, for S. tenacellus replacement of a small residue in position 254 by glutamine and Asn261 by aspartate was found to be the likely causes for resistance to (E)-beta-methoxyacrylates. The latter exchange is also found in Schizosaccharomyces pombe, which we found also to be naturally resistant to (E)-beta-methoxyacrylates.

Amino Acid Sequence↗

PubMed2002

A molecular approach for early monitoring of decay basidiomycetes in wood chips.

A DNA-based method for the early detection and identification of decay basidiomycetes on wood chips is described. From 26 species of wood-decaying basidiomycetes and 20 species of wood-inhabiting ascomycetes, DNA fragments containing the internal transcribed spacers, 5.8S and partial 28S rDNA were amplified by PCR using the ITS1-F-NL2 rDNA primer pair and analysed through sequencing and restriction digestion. A decay basidiomycete-specific restriction fragment length polymorphism (RFLP) pattern was generated by DraI. Using this PCR-RFLP method the detection of decay fungi is possible 4 days after the inoculation of the wood chips with either a single culture or mixed species.

Basidiomycota↗