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Analyses of soil fungal communities in adjacent natural forest and hoop pine plantation ecosystems of subtropical Australia using molecular approaches based on 18S rRNA genes.

Soil fungal communities were studied using 18S rDNA-based molecular techniques. Soil DNA was analyzed using temperature gradient gel electrophoresis (TGGE), single-stranded conformational polymorphism (SSCP), cloning and sequencing methods, following community DNA extraction and polymerase chain reaction (PCR). The extracted community DNA was successfully amplified using the primer pair of EF4f-Fung5r which produced ca. 550bp 18S rDNA fragments. TGGE screening of the PCR products showed some differences in band position and intensity between two soil samples in adjacent natural forest (YNF) and hoop pine plantation (YHP) ecosystems at Yarraman in subtropical Australia. TGGE and SSCP could be used for screening PCR products. However, care must be exercised when interpreting the TGGE and SSCP results with respect to microbial diversity, because one band may not necessarily represent one species. It is recommended that the PCR products should be purified before TGGE or SSCP screening. SSCP screening of the clone sequences revealed differences among the clones. Sequence and phylogenetic analyses revealed that all obtained clones were affiliated to the kingdom Fungi, including three phyla, i.e., Zygomycota, Ascomycota and Basidiomycota. Our results suggested that community DNA extraction, PCR, cloning, SSCP screening of clones, sequencing of selected clones and phylogentic analyses could be a good strategy in investigation of soil fungal community and diversity.

Ascomycota↗

The mitochondrial apocytochrome b genes of two Agrocybe species suggest lateral transfers of group I homing introns among phylogenetically distant fungi.

The Agrocybe chaxingu and Agrocybe aegerita mitochondrial apocytochrome b coding sequences are highly similar (97% of nt identity), but have highly different sizes (2312 and 4867nt, respectively), due to the presence of three large group IB introns: two (iAae1 and iAae2) in A. aegerita, one (iAch1) in A. chaxingu. All these introns encode a homing endonuclease (HE) similar to those described in introns of mitochondrial genes (cob, cox1, and nad5) from various organisms. Phylogenetic trees were built with these HE sequences. From these trees, the Agrocybe coding introns argue for recent lateral transfers, i.e., occurring after the separation of the two Agrocybe species, involving phylogenetically distant fungi such as members of the Ascomycota phylum (for iAch1 and iAae2) and, for the first time to our knowledge, a member of the Chytridiomycota phylum (for iAae1). The grouping of the HE gene (HEG) sequences according to the mitochondrial gene (cob, cox1, and nad5) where they are inserted, suggests modifications of the interactions between the HE and the recognized sequences, leading to new target genes. The largest distribution of the iAch1 HE, shared by several cob and cox1 mitochondrial genes from Ascomycota, Basidiomycota, and Chytridiomycota phyla, suggests a higher target flexibility of this HE, perhaps related to the presence of two different LAGLIDADG motifs in the catalytic site of the enzyme.

Agaricales↗

Genetics of self/nonself recognition in Serpula lacrymans.

This study provides an analysis of the vegetative incompatibility system in Serpula lacrymans (Basidiomycota), a genetic system used to recognize nonself in fungi. Seventy-five worldwide isolates could be grouped into eight vegetative compatibility (VC) types, some of them distributed on different continents. Mating studies combined with vegetative incompatibility analyses revealed that the vegetative incompatibility response between isolates mainly could be explained by two biallelic vegetative incompatibility (vic) loci. The frequency distributions of the interpreted vic alleles do not seem to support the idea of frequency-dependent or balancing selection acting on the vic loci. We find little genetic variation at the vic loci and in one of the loci there was a significant heterozyote deficiency among strains in the overall material. The results may be explained by a recent worldwide dispersal of a few S. lacrymans isolates and, correspondingly, only a few vic alleles are being maintained in these populations.

Basidiomycota↗

Cantharellus pleurotoides, a new and unusual basidiomycete from Guyana.

Cantharellus pleurotoides sp. nov. (Cantharellaceae, Cantharellales, Basidiomycota) is described from the Pakaraima Mountains of Guyana, occurring in rainforests dominated by ectomycorrhizal Dicymbe spp. (Caesalpiniaceae). This fungus is singular among Cantharellus species described worldwide in possessing a pleurotoid basidioma. Macromorphological, micromorphological, and habitat data are provided for the new species.

Basidiomycota↗

Marine fungi from Mira river salt marsh in Portugal.

The occurrence of fungi in the Mira salt marsh, Portugal was investigated for 12 months. Baits of Spartina maritima stems were exposed to permanent or temporary submersion at the upper and lower limits of the intertidal zone. The baits were observed for fruit bodies and spores directly and after incubation in moist chambers. Twenty six marine species were identified (17 Ascomycota, two Basidiomycota and seven mitosporic fungi). Twenty four are new records for Portugal. Nia globospora Barata and Basílio was published as a new species. Species were characterized with respect to frequency of occurrence, colonization capability and substrate succession. The diversity and similarity indexes of the fungi under different conditions were determined.

Ascomycota↗

Parenthesome structure of some corticioid fungi.

The parenthesome structure of seven corticioid species, traditionally referred to the family Corticiaceae (Basidiomycota), were studied in order to better understand their taxonomic position: Phanerochaete velutina, Phlebia radiata, P. rufa, Rhizochaete americana (syn. Ceraceomyces americana), R. brunnea, R. filamentosa (syn. Phanerochaete filamentosa) and R. radicata (syn. Phanerochaete radicata). All possessed the perforate type of parenthesome that is commonly encountered in homobasidiomycetes. This feature excludes the above taxa from both the hymenochaetoid and the cantharelloid clades which are the only groups that have imperforate parenthesomes in the homobasidiomycetes.

Basidiomycota↗

Ectomycorrhizal mycobionts of Pisonia grandis on coral cays in the Capricorn-Bunker group, Great Barrier Reef, Australia.

The diversity of ectomycorrhizal mycobionts of Pisonia grandis (Nyctaginaceae) from coral cays in the Capricorn-Bunker group, Great Barrier Reef, Australia, was examined. Only two ectomycorrhiza morphotypes (brown and black) were identified in soil from seven cays and DNA from both morphotypes was subjected to ITS-RFLP and sequence analysis. The brown morphotype was present in soil from all cays but the black morphotype was only observed in soil from three cays. ITS-RFLP analysis showed that the brown and black morphotypes were formed by different fungal taxa, with the RFLP pattern for the black morphotype being consistent with that of the culture previously obtained from black ectomycorrhizal roots on Heron Island. Comparison with the GenBank database revealed that closest matches to both morphotypes were sequences for various Thelephoraceae (Basidiomycota), but the brown and black morphotypes had only 80% sequence similarity to each other. Neighbour-joining analysis of these sequences with sequences for other Thelephoraceae grouped the brown and black morphotypes in a well-supported clade with several Tomentella species, suggesting that both belong to this genus. The data are discussed in relation to ectomycorrhizal fungal diversity and the coral cay habitat.

Animals↗

New PCR assay detects rare tooth fungi in wood where traditional approaches fail.

Lu et al. (2002) described a method for identifying Hericium species by PCR, using the primers HT-U1 and HT-L1 which they specifically designed for this purpose. In our hands these primers do not appear to discriminate between tooth fungi and other wood decay species. Therefore PCR primers were designed that discriminated Creolophus cirrhatus from other species (HER2F/HER3R), and which discriminate Hericium alpestre, H. coralloides and H. erinaceus from other wood decay Ascomycota and Basidiomycota but not from each other (HER2F/HER2R). Using the HER2F/HER3R primers together with traditional isolation and direct incubation procedures, the location of C. cirrhatus in Turkey oak logs was mapped. The PCR approach often detected C. cirrhatus in locations where it was suspected to be, based on patterns of staining and decay, but where it was not revealed by isolation onto agar media, emphasising the value of adopting several approaches to unravel fungal community structure in wood.

Basidiomycota↗

Patterns of interaction between Populus Esch5 and Piriformospora indica: a transition from mutualism to antagonism.

Piriformospora indica (Sebacinaceae, Basidiomycota) is an axenically cultivable, plant growth promoting root endophyte with a wide host range, including Populus. Rooting of Populus Esch5 explants started within 6 days after transfer to WPM medium. If such plantlets with roots were inoculated with P. indica, there was an increase in root biomass, and the number of 2nd order roots was increased significantly. A totally different observation was recorded when the explants were placed into WPM with pre-grown P. indica. The interaction led to complete blocking of root production and severely inhibited plant growth. Additionally, branched aerial roots appeared which did not penetrate the medium. On contact with the fungal colony or the medium, the ends of the aerial roots became inflated. Prolonged incubation stimulated the fungus to colonize aerial parts of the plant (stem and leaves). Mycelium not only spread on the surface of the aerial parts, but also invaded the cortical tissues inter- and intracellularly. Detached Populus leaves remained vital for 4 - 5 weeks on sterile agar media or on AspM medium with pre-grown P. indica. When the fungus was pre-grown on culture media such as WPM, containing ammonium as the main source of nitrogen, leaves in contact with the cultures turned brownish within 4 - 12 h. Thereafter, the leaves bleached, and about one day later had become whitish. Thus, cultural conditions could alter the behaviour of the fungus drastically: the outcome of the interaction between plant and fungus can be directed from mutualistic to antagonistic, characterized by fungal toxin formation and extension of the colonization to Populus shoots.

Basidiomycota↗

The root endophytic fungus Piriformospora indica requires host cell death for proliferation during mutualistic symbiosis with barley.

Fungi of the recently defined order Sebacinales (Basidiomycota) are involved in a wide spectrum of mutualistic symbioses (including mycorrhizae) with various plants, thereby exhibiting a unique potential for biocontrol strategies. The axenically cultivable root endophyte Piriformospora indica is a model organism of this fungal order. It is able to increase biomass and grain yield of crop plants. In barley, the endophyte induces local and systemic resistance to fungal diseases and to abiotic stress. To elucidate the lifestyle of P. indica, we analyzed its symbiotic interaction and endophytic development in barley roots. We found that fungal colonization increases with root tissue maturation. The root tip meristem showed no colonization, and the elongation zone showed mainly intercellular colonization. In contrast, the differentiation zone was heavily infested by inter- and intracellular hyphae and intracellular chlamydospores. The majority of hyphae were present in dead rhizodermal and cortical cells that became completely filled with chlamydospores. In some cases, hyphae penetrated cells and built a meshwork around plasmolyzed protoplasts, suggesting that the fungus either actively kills cells or senses cells undergoing endogenous programmed cell death. Seven days after inoculation, expression of barley BAX inhibitor-1 (HvBI-1), a gene capable of inhibiting plant cell death, was attenuated. Consistently, fungal proliferation was strongly inhibited in transgenic barley overexpressing GFP-tagged HvBI-1, which shows that P. indica requires host cell death for proliferation in differentiated barley roots. We suggest that the endophyte interferes with the host cell death program to form a mutualistic interaction with plants.

Apoptosis↗

Chronic maxillary sinusitis associated with the mushroom Schizophyllum commune in a patient with AIDS.

Invasive infection with fungi of the Basidiomycota (rusts, smuts, toadstools, mushrooms, and puffballs) is extremely rare. We report such an infection in a patient with human immunodeficiency virus disease who presented with chronic maxillary sinusitis associated with the mushroom Schizophyllum commune. The organism was isolated from the surgical drainage material, and septate hyphae were seen invading the maxillary submucosa. The limited literature on this subject is reviewed.

Acquired Immunodeficiency Syndrome↗

Xylose reductase from the Basidiomycete fungus Cryptococcus flavus: purification, steady-state kinetic characterization, and detailed analysis of the substrate binding pocket using structure-activity relationships.

Xylose reductase has been purified to apparent homogeneity from cell extracts of the fungus Cryptococcus flavus grown on D-xylose as carbon source. The enzyme, the first of its kind from the phylum Basidiomycota, is a functional dimer composed of identical subunits of 35.3 kDa mass and requires NADP(H) for activity. Steady-state kinetic parameters for the reaction, D-xylose + NADPH + H(+)<--> xylitol + NADP(+), have been obtained at pH 7.0 and 25 degrees C. The catalytic efficiency for reduction of D-xylose is 150 times that for oxidation of xylitol. This and the 3-fold tighter binding of NADPH than NADP(+) indicate that the enzyme is primed for unidirectional metabolic function in microbial physiology. Kinetic analysis of enzymic reduction of aldehyde substrates differing in hydrophobic and hydrogen bonding capabilities with binary enzyme-NADPH complex has been used to characterize the substrate-binding pocket of xylose reductase. Total transition state stabilization energy derived from bonding with non-reacting sugar hydroxyls is approximately 15 kJ/mol, with a major contribution of 5-8 kJ/mol made by interactions with the C-2(R) hydroxy group. The aldehyde binding site is approximately 1.2 times more hydrophobic than n-octanol and can accommodate linear alkyl chains of <or=6 carbons. Hydrophobic interactions provide a total binding energy of approximately 10 kJ/mol. Specificity for the aldehyde substrate is achieved through large decreases in apparent K(m) ( approximately 100-fold) and smaller but significant increases in turnover number ( approximately 5-fold). We observed up to 250-fold preference of xylose reductase for reaction with pyridine carbaldehydes, 4-nitro-benzaldehyde, and alpha-oxo-aldehydes over reaction with D-xylose, perhaps reflecting a secondary role of this enzyme in detoxication metabolism of reactive endogenous aldehydes and compounds of xenobiotic origin.

Aldehyde Reductase↗

Bannoa hahajimensis gen. nov., sp. nov., and three related anamorphs, Sporobolomyces bischofiae sp. nov., Sporobolomyces ogasawarensis sp. nov. and sporobolomyces syzygii sp. nov., yeasts isolated from plants in Japan.

Fourteen ballistoconidium-forming yeast strains were isolated from leaves of plants collected in the Ogasawara Islands, which are isolated islands in the Pacific Ocean, about 1,000 km south of the Japanese mainland, in the subtropical zone. The 14 isolates were characterized by the absence of xylose in whole-cell hydrolysates, the presence of Q-10(H2) as the major ubiquinone isoprenologue, G+C contents of 47.6-52.0 mol%, the inability to ferment sugars or to assimilate nitrate and positive Diazonium blue B and urease reactions. They formed a phylogenetically coherent cluster within the Erythrobasidium lineage in the Urediniomycetes of the Basidiomycota based on 18S rDNA sequences. Analyses of the nucleotide sequences of internal transcribed spacer regions and DNA complementarity showed that four genospecies were recognized among the 14 isolates. A mating reaction was observed in one of the four genospecies, which produced one-celled basidia on dikaryotic hyphae with clamp connections. On the basis of the morphological, physiological, chemotaxonomic and phylogenetic analyses, a new teleomorphic genus, Bannoa, is proposed, in which one novel species is described, Bannoa hahajimensis gen. nov., sp. nov. (type strain OK-248(T) = JCM 10336T = CBS 9039(T)). The other three anamorphic genospecies are described as Sporobolomyces bischofiae sp. nov. (type strain OK-257T = JCM 10338T =CBS 9041T), Sporobolomyces ogasawarensis sp. nov. (type strain OK-14T = JCM 10326T = CBS 9038T) and Sporobolomyces syzygii sp. nov. (type strain OK-227T = JCM 10337T = CBS 9040T.

Basidiomycota↗

Novel anamorphic mite-associated fungi belonging to the Ustilaginomycetes: Meira geulakonigii gen. nov., sp. nov., Meira argovae sp. nov. and Acaromyces ingoldii gen. nov., sp. nov.

Three novel mite-associated basidiomycetous species are described in two new anamorph genera as Meira geulakonigii gen. nov., sp. nov. (type CBS 110052(T)=NRRL Y-27483(T)=AS 004(T)), Meira argovae sp. nov. (type CBS 110053(T)=NRRL Y-27482(T)=AS 005(T)) and Acaromyces ingoldii gen. nov., sp. nov. (type CBS 110050(T)=NRRL Y-27484(T)=AS 001(T)). Morphologically, these fungi are similar to the yeast-like fungi classified in the Ustilaginales, such as Pseudozyma species. However, analysis of the D1/D2 domain of the LSU rDNA suggests that they belong to two different lineages within the Exobasidiomycetidae of the Ustilaginomycetes (Basidiomycota). Furthermore, these fungi may be of interest for the biocontrol of mites, as they reduced mite numbers by approximately 80 % after inoculation.

Animals↗

Curvibasidium cygneicollum gen. nov., sp. nov. and Curvibasidium pallidicorallinum sp. nov., novel taxa in the Microbotryomycetidae (Urediniomycetes), and their relationship with Rhodotorula fujisanensis and Rhodotorula nothofagi.

Strains of Rhodotorula fujisanensis (Basidiomycota, Urediniomycetes, Microbotryomycetidae), including the type strain, are sexually compatible and produce clamped mycelium with teliospores. However, as teliospore germination had not been documented, the complete sexual cycle was not known. During the course of this work, the basidial stage of R. fujisanensis was characterized. In addition, mating studies employing isolates that were identified preliminarily as Rhodotorula nothofagi, a species that is related closely to R. fujisanensis, yielded mycelium with teliospores, which formed basidia and basidiospores. The new data were evaluated by using several criteria, including the available molecular phylogenetic framework for the Microbotryomycetidae. Curvibasidium gen. nov. is described here, to accommodate two teleomorphs: Curvibasidium cygneicollum sp. nov. (CBS 4551T), which is described as the sexual stage of R. fujisanensis, and Curvibasidium pallidicorallinum sp. nov. (CBS 9091T), which is related closely to R. nothofagi, but does not represent its sexual stage.

Basidiomycota↗

Molecular tools for the identification of ectomycorrhizal fungi--taxon-specific oligonucleotide probes for suilloid fungi.

Five taxon-specific oligonucleotide probes are described that can be used to help identify the fungal components of ectomycorrhizae. Comparisons among partial sequence from the mitochondrial large subunit rRNA gene (mt-LrRNA) were used to select the probes, which were intended to be specific to several taxa within the suilloid group of the Boletales (Basidiomycota). Probes S1, R1, and G1 were targeted at the genera Suillus, Rhizopogon and Gomphidius; probe G2 was designed to recognize the family, Gomphidiaceae, and probe US1 was designed to recognize all of these taxa and any other members of the suilloid group. The specificity of each probe was determined empirically by testing their ability to hybridize to PCR amplified fragments derived from 84 species of basidiomycetes. Although none of the probes exhibited their intended specificity, all specifically hybridized to useful subsets of taxa, and collectively they can be used to identify many suilloid taxa to the generic level or below. The probes were also tested for their ability to identify field collected mycorrhizae and were found to perform well.

Base Sequence↗

Multiple gene genealogies and AFLPs suggest cryptic speciation and long-distance dispersal in the basidiomycete Serpula himantioides (Boletales).

Serpula himantioides (Boletales, Basidiomycota) produces thin resupinate basidiocarps on dead coniferous wood worldwide and causes damage in buildings as well. In this study, we present evidence for the existence of at least three phylogenetically defined cryptic species (referred to as Sib I-III) within the morphospecies S. himantioides, a conclusion based on analyses of sequence data from four DNA regions and amplified fragment length polymorphisms (AFLPS). A low degree of shared sequence polymorphisms was observed among the three lineages indicating a long-lasting separation. The AFLPs revealed two additional subgroups within Sib III. Results from mating studies were consistent with the molecular data. In Sib III, no correspondence between genetic and geographical distance was observed among isolates worldwide, presumably reflecting recent dispersal events. Our results indicate that at least two of the lineages (Sib II and Sib III) have wide sympatric distributions. A population genetic analysis of Sib III isolates, scoring sequence polymorphisms as codominant SNP markers, indicates that panmictic conditions exist in the Sib III group. This study supports the view that cryptic speciation is a common phenomenon in basidiomycete fungi and that phylogenetic species recognition can be a powerful inference to detect cryptic species. Furthermore, this study shows that AFLP data are a valuable supplement to DNA sequence data in that they may detect a finer level of genetic variation.

Basidiomycota↗

Cryptotrichosporon anacardii gen. nov., sp. nov., a new trichosporonoid capsulate basidiomycetous yeast from Nigeria that is able to form melanin on niger seed agar.

Five yeast isolates obtained from cashew tree flowers in Nigeria resembled Cryptococcus neoformans phenotypically by producing brown pigmented colonies on niger seed agar, expressing a capsule, and being able to grow at 37 degrees C. However, rRNA gene sequences, including the 18S rRNA gene, the D1/D2 domains of the 26S rRNA gene and the ITS1+2 regions, suggested that these yeasts form a basal lineage within the Trichosporonales (Tremellomycetidae, Hymenomycetes, Basidiomycota, Fungi). Since the isolates could not be identified with any known genus and species within the Trichosporonales, we describe them as Cryptotrichosporon anacardii gen. et sp. nov. with CBS 9551(T) (=NRRL Y-27671) as the type strain. The taxonomic conflict between phenetic and molecular classification schemes within this group of fungi is discussed, and is resolved in favor of the latter.

Agar↗