The phylogenetic relationship of 52 specimens representing 30 species of Anthracoidea (Ustilaginales) was investigated by molecular analyses using sequence data from the large subunit (LSU) of nuclear rDNA. Phylogenetic trees were inferred with neighbour-joining (NJ), maximum parsimony (MP), and Bayesian Markov chain Monte Carlo (MCMC) methods. The results are discussed with respect to the species concept and the subdivision of the genus into subgenera and sections. Collections from different hosts and localities were compared. Our analyses can neither support nor significantly reject the hypothesis of the bipartition of the genus Anthracoidea. Thus, the representatives of the subgenus Proceres appeared in the NJ analysis as a moderately supported monophylum, whereas MCMC analysis revealed a polyphyletic topology for this group. Paraphyly of the subgenus Anthracoidea was supported by all methods used. Sections Echinosporae and Leiosporae were each represented by two species in our analyses which grouped together with high support. Section Anthracoidea should be restricted to a highly supported group with extremely irregular to angular teliospore shape. However, these three sections do not cover the whole diversity of the subgenus Anthracoidea. Molecular data largely supported the traditional circumscription of species, and species delimitations are discussed.

The mountain pine beetle, Dendroctonus ponderosae, and its fungal associates are devastating the lodgepole pine forests in British Columbia, Canada. During our fungal survey, an unknown Leptographium species has been consistently isolated from both D. ponderosae and infested lodgepole pine (Pinus contorta var. latifolia). This Leptographium species has similar morphology with the Leptographium anamorph of Ophiostoma clavigerum whose association with the D. ponderosae is well known. However, thorough morphological comparisons showed that this fungus is distinct from all the other Leptographium species described in the literature, and especially from O. clavigerum. Comparison of DNA sequences of multiple loci and the profiles by the PCR-RFLP marker also confirmed that this Leptographium species represents an undescribed taxon. Based on its distinct morphological, physiological characteristics and phylogenetic position, we describe it as L. longiclavatum sp. nov.

14 different yeasts were isolated from the gut of a variety of insects, including beetles, lacewings, fishflies, craneflies, and a cockroach. One of the yeasts was found both in the gut and on the body surface of a beetle larva. Based on ribosomal DNA sequence comparisons and phenotypic characters, the yeasts were identified as Candida membranifaciens, C. tenuis, Pichia nakazawae, and nine undescribed taxa in Saccharomycotina. All the undescribed taxa reproduced only asexually, and they fit within the limits of the polyphyletic genus Candida. The new species and their type strains are Candida blattariae NRRL Y-27703T, C. amphixiae NRRL Y-27704T, C. michaelii NRRL Y-27705T, C. cerambycidarum NRRL Y-27706T, C. gorgasii NRRL Y-27707T, C. endomychidarum NRRL Y-27708T, C. temnochilae NRRL Y-27763T, C. sinolaborantium NRRL Y-27765T, and C. lessepsii NRRL Y-27766T spp. nov. Phylogenetic analysis of combined small and large subunit ribosomal DNA sequences placed C. amphixiae, C. michaelii, C. cerambycidarum, C. gorgasii, C. endomychidarum, and C. lessepsii in a statistically well supported clade with C. blattariae, C. membranifaciens, C. friedrichii, and C. buinensis as sisters to the clade. The other two new taxa, C. temnochilae and C. sinolaborantium, formed an independent clade basal to the major clade containing C. membranifaciens and closely related taxa. C. sinolaborantium occurred in both Panama and the USA, but there were genetic differences between the isolates from the two places.

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.

The shelled grains of glutinous broomcorn millet, Panicum miliaceum, were used as solid substrate to prepare granular cultures of Zoophthora radicans, an entomophthoralean biocontrol agent against numerous insect pests. Steamed millet grains were inoculated by mixing 15 g millet grains (D.W.) with mashed pieces of half a 60-mm-dish colony in 3 ml modified Sabouraud dextrose broth and incubated at 15 °C and L[ratio ]D 12[ratio ]12 for up to 24 d. 20 grains were sampled at 3 d intervals from day six onwards and individually assessed for their sporulation capacity using a self-designed device for spore collection. The millet cultures after [ges ]12 d incubation produced 12.0–14.9×104 spores grain−1 during a 7 d period. The maximal sporulation capacity associated with the 21 d-old culture was about half of that of Z. radicans-killed Plutella xylostella larvae (28.7×104 spores cadaver−1), which individually were at least three times larger than the millet grains. Based on the time–concentration–mortality responses of second-instar P. xylostella larvae to Z. radicans in three independent bioassays, the spores ejected from the cultured millet grains, from the mycelial mats from liquid culture, and from larval cadavers displayed insignificant variations in infectivity to the host species, and yielded similar LC50 and LT50 estimates. Conclusively, the millet-based technology for production of granular cultures of Z. radicans was easy, inexpensive and highly efficient, and it could be superior to previous methods used in mass production of mycelium-based preparations of Entomophthorales since this new approach requires no special additives, drying, freezing and milling. This technology may suit to mass production of culturable but nutritionally fastidious entomopathogens from the Entomophthorales.

The effects of grazing by the collembolan Folsomia candida on mycelial foraging patterns of Hypholoma fasciculare growing from beech (Fagus sylvatica) wood block inocula in trays of non-sterile soil was investigated. The wood inocula differed in size, state of decay (time for which wood has been colonized: 2 yr, 1 yr, 6 and 3 months) and nutrient status (inocula colonized on malt agar or nutrient agar). Mycelia were most luxuriant, had greater hyphal coverage and extended more rapidly from 2 yr old than younger inocula, from 4 cm3 than 1 cm3 inocula, and from inocula colonized on malt extract agar rather than on distilled water agar. Grazing dramatically reduced coverage and extension, especially in the less luxuriant systems characterized by many fine hyphae and fewer mycelial cords. Grazing by collembola often resulted in points of more rapid outgrowth as cords with a fanned margin. Results are discussed in terms of fungal foraging strategies.

To determine the relationship of Australian members of the genus Lepista with those from other parts of the world, genetic variation of isolates representing 27 accessions was assayed by cleaved amplified polymorphic sequences (CAPS) and random amplified polymorphic DNA (RAPD). CAPS and RAPD identified eight and seven groups, respectively. CAPS Groups 1 and 2 and RAPD Group 1 consisted of French and Australian accessions classified as morphospecies L. nuda. CAPS Group 3 and RAPD Groups 2 and 2A consisted of mostly Australian isolates identified as L. sordida var. sordida or L. sordida var. umbonata. Isolates earlier identified as morphospecies L. sp. were also placed in CAPS Group 3 and RAPD Group 2A indicating that these isolates are L. sordida var. sordida. In addition, three smaller groups were distinguished. A French isolate of L. sordida var. sordida was placed in distinctly separate CAPS and RAPD groups to Australian L. sordida var. sordida Groups 4,4 respectively. A French isolate of L. sordida var. aianthina was placed in CAPS and RAPD Groups 3,3. An accession of L. saeva was placed in CAPS Group 6 and RAPD Group 5, separate from other isolates. RAPD Groups 6 and 7 consist, respectively, of Greek and American accessions of L. nuda that were only distantly related to the Australian and French accessions of this morphospecies: CAPS also separated these isolates from each other and from all other isolates.

The data suggest that the classification of morphospecies and varieties within Lepista cannot be determined on the basis of morphology alone. The Greek and American accessions of L. nuda are separated from the French and Australian accessions and may not be L. nuda. Similarly, the Australian accessions currently classified as L. sordida var. sordida together with the accessions of L. sordida var. umbonata are distinct from the French accessions of L. sordida var. sordida suggesting that Australian isolates may represent a new species or variety. Further studies using a combination of morphological and genetic characterization are required to provide a more accurate understanding of Lepista species.

Macromycetes recorded from beds of creeping willow, Salix repens, in the northern archipelagos of Shetland and Orkney are listed and discussed. Two components are demonstrated, an arctic-alpine group and one which is more typical of lowland frondose woodland communities in mainland Britain. The importance of maintaining, even encouraging the development of creeping willow beds with their attendant ectomycorrhizal fungi in conservation terms are explored.

The ascomycetous genus Cordyceps accommodates endoparasitic species that attack arthropods or other fungi. Analyses of ITS nrDNA sequence data of 72 taxa from the teleomorph genera Cordyceps, Claviceps, Epichloë, and the anamorph genera Akanthomyces, Beauveria, Metarhizium, Hirsutella, Hymenostilbe, Paecilomyces, Polycephalomyces, and Tolypocladium assigned the taxa to four main evolutionary lineages not reflected in the current classification of Cordyceps. Ten subclades were recognized from separate analyses of data subsets. Judged from the ITS phylogenies, Cordyceps spp. with branched stromata were highly supported as a divergent lineage. Host specificity was found to be of limited phylogenetic significance, and several host shifts are suggested to have occurred during the evolution of Cordyceps. Similar ascospore morphology was not reflected in the phyletic groups, and closely related taxa showed large interspecific variation with respect to the number of segments in which the ascospores are divided. However, combinations of selected characters were found to delimitate some lineages, e.g. all Cordyceps spp. that attack hosts in the insect orders Coleoptera and Lepidoptera, and with non-immersed perithecia and clavate to acicular, brightly yellowish to reddish stromata, constituted a separate clade. Furthermore, all Cordyceps spp. with perithecia obliquely immersed in the stroma were recognized as a distinct monophyletic group. This clade is additionally characterized by the formation of anamorphs ascribable to the genus Hymenostilbe. The mycogenous Cordyceps spp. grouped in a separate subclade, interspersed by two cicadaen parasites and all Tolypocladium spp. except T. parasiticum. Tolypocladium and Beauveria were found to be polyphyletic. The included Claviceps and Epichloë taxa appeared to be derived within Cordyceps, thus making Cordyceps paraphyletic as suggested in other studies.

Rust fungi in the genus Phragmidium are frequent pathogens of both wild and cultivated roses. We investigated the occurrence and relationships of rusts on dog roses, Rosa sect. Caninae (Rosa canina, R. corymbifera and R. rubiginosa) in Germany. Two Phragmidium species, P. mucronatum and P. tuberculatum, were able to infect each of the three dog rose species examined. However, the overall infection of R. rubiginosa was significantly lower, which could be important for rose breeding. Despite overlapping host ranges, the evolutionary background of P. tuberculatum and P. mucronatum is quite distinct. Phylogenetic analyses of the D1/D2 region of the LSU rDNA suggest that P. mucronatum shares a common ancestor with other rose rusts, whereas P. tuberculatum evolved from a Rubus-Sanguisorba rust clade and must have undergone a host shift to Rosa spp.

Ericoid endomycorrhizal fungi (two isolates of Hymenoscyphus ericae obtained from unpolluted heathlands and two H. ericae-type endophytes isolated from Calluna vulgaris growing on Cu-contaminated mine spoil) were grown for 14 d on 10% Rorison's solution containing sodium phytate as the sole P source and either trace (0.16 μM) or elevated (0.25 mM) concentrations of Cu. The elevated levels of Cu in the medium had no effect on the growth of the two H. ericae-type endophytes from mine spoil sites but caused a significant reduction in growth of the two H. ericae isolates from unpolluted sites. Wall, cytoplasmic and extracellular fractions were assayed for phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) activity. Km and Vmax values varied between the different endophytes and both were highest in the wall fractions. Wall-bound phosphatase activity, excluding PDEase of one H. ericae-type endophyte, was generally unaffected after the isolates had been grown on medium containing 0.25 mM Cu. Extracellular PDEase of the two H. ericae-type endophytes from mine spoil sites was stimulated by 0.25 mM Cu in the growth medium. Cu concentrations up to 5.0 mM in the assay medium did not inhibit wall-bound phosphatase activity whereas three of the isolates showed a stimulation of extracellular activity with increasing Cu. The results are discussed in relation to phosphatase activity of ericoid endophytes on Cu-contaminated substrates.

Basidiomes of 43 samples of eight Amanita species were gathered from different habitats of Hungary. The mineral composition (22 elements) was analysed by the ICP method in three independent replications, and mineral compositions found as discussed and compared. The Amanita species analysed were very different in As-, Cd-, Cr-, Mo-, Mn-, Se- and mainly in V-content. Other elements (Al, B, Ba, Ca, Co, Cu, Fe, Mg, Na, Ni, Sr, Ti, Zn) occur in the basidiomes in balanced concentrations. The K and P contents have the lowest differences. Summarizing all the data (n=43), the average mineral status of species of Amanita can be deduced. The lowest variability measured was for K and P, and the highest for chromium, nickel and vanadium.

Specific, significant accumulation was found only for vanadium, due to the previously demonstrated occurrence of a binding molecule ‘amavadine’ in the basidiomes of A. muscaria. Remarkable Cd-levels were estimated in A. pantherina and A. muscaria (11.4 and 12.3 mg kg−1D.M., respectively). The higher contents of other elements (e.g. K, practically in all species; Se in A. strobiliformis) are analytical facts but, not accumulations.

The mineral compositions of the ectomycorrhizal genus Amanita, of litter decomposing Agaricus and of wood decaying Trametes were compared. Some significant differences were found (AsAmanita<AsAgaricus; CdAmanita<CdAgaricus; CuAmanita<CuAgaricus; PAmanita<PAgaricus; KAmanita>KTrametes; PAmanita>PTrametes) but it seems that the mineral composition of the basidiomes is practically independent of the ectomycorrhizal habit. The specificities of the fungi-tree symbiotic interactions are known, and well documented (higher uptake and transport of certain elements first of all of P), however, the differences found in the mineral components are due to other factors (e.g. substrates, accumulating ability) and not to the mycorrhizal status.

On nutrient-poor rock surfaces, yeast-like black fungi (also called microcolonial fungi, MCF) may derive organic carbon either from the atmosphere or from interactions with other rock-inhabiting microorganisms. Interactions between free-living rock inhabiting heterotrophic fungi and phototrophic algae were investigated using axenic cultures. Five typical MCF strains were incubated with pure cultures of four lichen photobionts isolated from lichens growing in similar locations. After 2–12 months of combined cultivation, the fungal and algal colonies developed an overlapping structure involving both partners. Microscopic, histological and ultrastructural methods were used to investigate the changes that occur after prolonged contact. Histological analysis demonstrated changes in the spatial organisation of mixed colonies when in contact with each other. The branching of fungal hyphae was more pronounced in the vicinity of algal cells, thus increasing the potential contact surface. Photobiont cells were not changed in size, but a reduction of sporulation could be observed in some Trebouxia strains. Close cell wall contacts between fungal and algal cells were formed and mucilage accumulated at the contact places. The ability of MCF to form associations with potential photobionts present on rock surfaces was thus demonstrated for pure cultures in vitro.

A new Hypocrella species with white pulvinate stromata collected in Puerto Rico and Costa Rica is described as H. zhongdongii sp. nov. Morphological and molecular evidence confirms that the new species of Hypocrella is the teleomorph of Aschersonia incrassata. It most closely resembles H. andropogonis; both A. incrassata and A. andropogonis are common yellow-spored species. The relationships of H. zhongdongii with other species in the genus are elucidated through phylogenetic analyses of three different genetic loci (LSU, RPB2, and mtSSU). Our analysis also sheds light on current subgeneric concepts in Aschersonia, in which the presence or absence of conidiomatal paraphyses is a major character to separate the genus into two subgenera. The present phylogenetic tree suggests that paraphyses have been lost or gained multiple times during evolutionary history, and do not define monophyletic groups.

Three polypeptides with manganese superoxide dismutase (MnSOD) activity were found in mycelium, zoospores and germinated cysts of Phytophthora nicotianae. Their relative molecular weights in non-denaturing gels were approximately 34.5, 36 and 50 kDa. No evidence for the presence of either iron or copper/zinc SODs was detected at any of the developmental stages examined. The level of activity of the MnSOD polypeptides was similar in mycelia and spores. Degenerate PCR was used to amplify partial genes of two different MnSODs, designated PnMnSOD1 and PnMnSOD2, from P. nicotianae. Southern blot analysis indicated that there are two PnMnSOD1 genes in the P. nicotianae genome. Full length sequence was obtained for one of these genes, PnMnSOD1a, from a P. nicotianae bacterial artificial chromosome (BAC) library. RNA blots probed with PnMnSOD1 showed similar levels of expression in vegetative and sporulating hyphae, lower levels in germinated cysts and no detectable expression in zoospores. PnMnSOD1a had 96%, 97% and 99% amino acid identity with homologous genes from P. ramorum, P. infestans and P. sojae, respectively. The second gene cloned from P. nicotianae, PnMnSOD2, had only 38% amino acid identity with PnMnSOD1a and was homologous to MnSODs that possessed an N-terminal mitochondrial targeting sequence in Phytophthora species and other eukaryotes. Southern blots indicated that there is one copy of PnMnSOD2 in the P. nicotianae genome. PnMnSOD2 was expressed at similar levels in mycelia and germinated cysts but PnMnSOD2 transcripts were not detectable in zoospores.

The anamorphic basidiomycete genus Marchandiomyces presently includes two common lichenicolous (lichen-inhabiting) species, M. corallinus and M. aurantiacus (teleomorph Marchandiobasidium aurantiacum). We describe here a new species, M. lignicola sp. nov., that is similar to M. corallinus in the colour of its sclerotia, but differs in having a wood-inhabiting (lignicolous) habit. The phylogenetic position of this lignicolous fungus was compared with the lichenicolous species of Marchandiomyces and related species currently placed in the basidiomycetous families Corticiaceae and Ceratobasidiaceae using parsimony, likelihood, and Bayesian analyses of complete sequences of the nuclear small subunit and internal transcribed spacers ribosomal DNA, and a portion of the nuclear large subunit ribosomal DNA. These DNA sequences were obtained from isolated cultures of freshly collected specimens. Significant Bayesian posterior probabilities, as well as maximum likelihood and parsimony analyses, indicate that the new lignicolous species is closely related to M. corallinus, the type species of Marchandiomyces. In most analyses these two species are monophyletic with the lichenicolous M. aurantiacus, although this relationship is not strongly supported. Since M. lignicola is more closely related to M. corallinus than to M. aurantiacus, either a transition to the lignicolous habit occurred recently within an ancestral lichenicolous group or, more likely, transition to the lichenicolous habit arose recently and in parallel from an ancestral lignicolous habit.

The entomophthoralean fungi Batkoa sp., Furia sp. and Neozygites floridana have been suggested for biocontrol of insect pests: the first two for control of spittlebug pests of pasture and sugarcane, and the third for mites of agricultural importance. To develop these agents as biopesticides and bioacaricides, it is important to have available culture media that maximize production at low cost. The research reported here evaluates, in different combinations and concentrations, the effect of four complex sources of nitrogen on production of mycelium or hyphal bodies in liquid media of all three species. Yeast extract allowed the highest production of Batkoa sp., with a concentration of 0.5% being the most suitable for vegetative (mycelial) growth. The combination of 0.33% each of yeast extract+beef extract+skim milk allowed the highest production of Furia sp. Mycelium. The combination of yeast extract+skim milk (0.5% of each) allowed the second highest production of Furia sp., and was the most suitable for mass production due to the lower cost. The combination of 1%each of yeast extract+peptone+skim milk was the most suitable for production of N.floridana hyphal bodies.

Cordyceps unilateralis, a fastidious insect pathogenic fungus, was found to produce red naphthoqinones, which possess anti-malarial activity. A method for the isolation and in vitro cultivation of this fungus was, therefore, developed in this study. Ascospores from ant cadavers were used as starting materials, and their secondary spores were allowed to germinate first on PDA for 2 d. Secondary spores were then transferred to Grace's Insect Cell Medium+2 mM glutamine and 10% fetal bovine serum, incubated for 7 d, after which the formation of mycelia was stimulated by transfer to potato dextrose broth. As a result, the growth of C. unilateralis from the steps of secondary spore germination, blastospore formation and mycelial growth was successfully completed for the first time.

Records of the Piptocephalidaceae and Chaetocladiaceae from over 800 samples of dung collected worldwide are analysed. Piptocephalis species were frequent, occurring especially on dung of rabbit (31% of all samples) in temperate regions. Chaetocladium and Syncephalis species were much less frequent. The status of these fungi as coprophils and reasons for the differences in distribution and substrate preference are discussed.