The influences of temperature on the fruit body induction of Flammulina velutipes were investigated. The fruit body could be induced after reducing of the ambient temperature under complete darkness, but could not be induced after light irradiation without temperature reduction. Fruit bodies formed under complete darkness elongated without pileus formation (pinhead fruit body). Protein expression patterns were investigated by two-dimensional electrophoresis, and 22 protein spots were found to be newly expressed in the mycelia and fruit bodies after reducing of the ambient temperature, but few protein spots were newly expressed in the mycelia after light irradiation without temperature reduction. Four of the spots had identical N-terminal sequences to polypeptides, Pf1, Pf3, Pfd3, and Pf6 which are previously sequenced as polypeptides expressed specifically during fruit body formation of F. velutipes.

Ceratocystis polonica and C. laricicola are two morphologically similar species that occur on conifers and reside in the Ceratocystis coerulescens species complex. They, however, represent two ecologically distinct entities. C. polonica causes blue stain on Norway spruce (Picea abies) and other spruce species (Picea spp.) in Eurasia and is associated with the bark beetles Ips typographus, I. typographus japonicus, I. amitinus and I. duplicatus. In contrast, C. laricicola lives in a symbiotic relationship with the bark beetles Ips cembrae and I. subelongatus that infest various larch species (Larix spp.). The objective of this study was to consider the phylogenetic relationships of C. polonica and C. laricicola and more specifically to determine the identity of Japanese isolates from both spruce and larch, based on sequences derived from the ITS regions of the rRNA operon, the β-tubulin gene and the HMG box of the MAT-2 gene. Isolates were also compared based on morphology and cultural characteristics. Comparisons of anamorph and teleomorph structures confirmed that C. polonica and C. laricicola are indistinguishable based on morphology. Both species had an optimal growth temperature of 25 °C. However, at temperatures between 31–33 °, C. polonica isolates grew slowly or not at all, while C. laricicola isolates grew more actively at these temperatures. Thus, a growth test at 32 ° can differentiate these species. Phylograms generated using parsimony for the three gene regions were strongly congruent. These showed three distinct clades supported by high bootstrap values. Two of the clades clearly separate C. laricicola from Europe and C. polonica, supporting the view that they represent two discrete taxa. A third clade included isolates obtained from galleries of Ips subelongatus on Larix kaempferi in Japan. This fungus clearly represents a discrete taxon that is closely related to, but distinct from C. laricicola, which is described here as C. fujiensis sp. nov.

Assessment of genetic and phenotypic diversity is necessary to confidently distinguish genotypes of Pleurotus eryngii when seeking traits of interest and to identify strains with high yield potential. We studied 154 strains from Italy for quantitative (shape, size and yield), qualitative (colour, malformations and growing behaviour), and molecular (RAPD and minisatellite) traits. This population consisted of isolates mostly belonging to P. eryngii var. eryngii, var. ferulae or var. nebrodensis, from different regions of Italy. A replicated cultivation trial, with three blocks and three replicates for each strain within the block, was used as an experimental design to calculate trait estimates. Significant differences were observed between strains for basidiome number and weight, while no significant differentiation for quantitative morphological traits was observed between geographical origins and taxonomic groups. Qualitative morphological traits were efficient in differentiating isolates of P. eryngii var. nebrodensis. On average, yield per strain (basidiome weight) was correlated more with basidiome number than with size. The most stable yield traits were basidiome number and weight per strain. An average heritability of 0.31 was estimated for yield related traits. A significant difference between var. ferulae and var. eryngii populations was detected for basidiome production measured as ‘average harvest time’. Molecular markers showed a high level of heterogeneity within populations and a low, but significant, degree of differentiation among populations defined a priori. No population-specific marker was detected and the differential pattern of variation between vars. ferulae and eryngii was due to frequency-dependent alleles. The nebrodensis type was more differentiated from var. eryngii than from var. ferulae using either molecular or qualitative morphological traits.

The phylogenetic relationships of two Meliola spp., one Meliolina sp., and 36 other ascomycetes were inferred from sequences of the nuclear small subunit ribosomal RNA gene. Phylogenetic analyses revealed that the Meliolaceae are members of the unitunicate Pyrenomycetes, and are not closely related to the bitunicate ascomycetes. The Meliolaceae have been hypothesised to be closely related to the Erysiphaceae, but the cladistic analyses showed that the two families are not closely related. The phylogenetic position of Meliolina is found amongst the bitunicate ascomycetes making it distantly related to the Meliolaceae.

To obtain an understanding of the phylogenetic relationships among the Georgefischeriales, septation, cellular interactions, teliospores, basidia, cultures and nucleotide sequences from the 5′ terminal domain of the nuclear large subunit rRNA gene were studied. Analyses of both morphological and molecular characters yield similar phylogenetic conclusions. The Georgefischeriales are divided into three groups, corresponding to the Eballistraceae, Georgefischeriaceae, and Tilletiariaceae. The basal dichotomy is between the Eballistraceae and the branch uniting the Georgefischeriaceae and Tilletiariaceae. The Tilletiariaceae are phragmobasidiate, whereas the Eballistraceae and the Georgefischeriaceae are holobasidiate. The Eballistraceae differ from the Georgefischeriaceae and Tilletiariaceae in the lack of the ballistospore mechanism. The systematic position of Tilletiopsis minor is unclear. The Eballistraceae, Eballistra and Phragmotaenium are proposed as new taxa. The descriptions of the Tilletiariaceae and Jamesdicksonia are emended. Except for Entyloma majus, E. parvum, Georgefischeria, Jamesdicksonia brunkii, J. obesa, Tilletiaria anomala, and Tolyposporella chrysopognis, the teleomorphic species of the Georgefischeriales are presented as new combinations.

Dactylaria tunicata sp. nov. from submerged wood in a freshwater stream from Australia is described and illustrated. It differs from all previously described species of Dactylaria in having uniseptate conidia with a hyaline sheath. The 41 species of Dactylaria accepted by de Hoog are listed, together with a further 41 validly published species of Dactylaria post de Hoog. A taxonomic key to 37 of the 41 post de Hoog species is provided, based on the literature, together with a composite illustration of their conidial morphology. The other four species are discussed, but they are not considered to be Dactylaria species (sensu de Hoog).

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.

A protocol for the generation of high yields of viable protoplasts has been developed for several highly branched (colonial) strains of the Quorn® myco-protein fungus, Fusarium graminearum A3/5. Driselase was found to produce higher protoplast yields (ca 109 g−1 wet weight) than the other lytic enzymes tested (Glucanex, Novozyme, β-glucuronidase, Sigma lytic enzyme, or ICN lytic enzyme), although yields differed for the various strains. Protoplast regeneration frequencies of 25–50% were observed when glucose (1·0 M) or sucrose (1·0 M) was used as the osmotic stabilizer. A highly branched strain of F. graminearum CC1-5, which grows better in submerged culture than the more sparsely branched wild-type strain (A3/5) was transformed using the hygromycin B resistance plasmid pAN7-1.

In response to the need for a faster, more reliable method for identifying Phytophthora cinnamomi in cork oak soils in Portugal, a simple, fast, sensitive molecular identification method is described. It is based on a colorimetric assay which involves an oligonucleotide capture probe covalently immobilised on microtitration wells, a multi-biotinylated oligonucleotide detection probe and the PCR-amplified target DNA. The target DNA is a 349 bp DNA fragment partially covering the 3′-translated and 3′-untranslated regions of the cinnamomin gene. When the specificity of the PCR reaction was evaluated in vitro using isolates of P. cinnamomi and eight other Phytophthora species, including the related P. cambivora, it was specific to P. cinnamomi. When 30 isolates of P. cinnamomi from oak roots in southern Portugal were assayed, 26 gave a strong positive response. The assay has a sensitivity of about 2–5 genome equivalents of P. cinnamomi. The reason for the negative response of four isolates remains unclear.

The growth of three wood inhabiting basidiomycetes (Microporus xanthopus, Pycnoporus sanguineus and Schizophyllum commune) isolated from a tropical coastal forest of Papua New Guinea is characterized in relation to temperature and NaCl salinity. P. sanguineus and M. xanthopus present a very similar response to NaCl salinity, but P. sanguineus has a wider tolerance for temperature variations. S. commune withstands wide temperature variations, but it is the only one to present a marked salt resistance. This correlates with its abundance along the sea. The presence of P. sanguineus in the same habitat is rather unexpected since it is very sensitive to NaCl salinity. Its high growth rate probably allows it to take advantage of periods of high rains that limit salt accumulation.

The identification of plant-associated isolates of Aphanomyces spp. has been mainly based on morphological characters. These types of features, however, can be unreliable due to their high variation and degree of overlap between different taxa. In this work, strains of Aphanomyces, with plant pathogenic and non-pathogenic characteristics, derived mainly from the roots of several leguminous crops in Sweden, as well as a number of reference strains of A. euteiches, A. cladogamus and A. cochlioides were compared by morphology, AT-rich DNA RFLPs and sequence analysis of the total ITS/5.8S rRNA gene region. Parsimony analysis of the sequence data of the total ITS region of 21 strains separated the taxa at the specific level, while two intra-specific groups were also detected within A. euteiches. One comprised all pathogenic isolates of A. euteiches from Sweden, together with two isolates from France and the USA. These isolates formed a homogenous group with identical ITS regions and a single AT-rich DNA RFLP banding pattern. The second ITS subspecific group included reference strains of A. euteiches with preferential pathogenicity to bean and alfalfa, all of which originated from the USA. The bean and alfalfa pathotypes were further distinguished by their different AT-rich DNA RFLPs. The non-pathogenic isolates of Aphanomyces from Sweden associated with the roots of several legumes, formed a genotypically uniform group with an identical ITS sequence and a single AT-rich DNA RFLP pattern. The ITS sequence of this group was identical to that of reference strains of A. cladogamus, suggesting that the non-pathogenic isolates belonged to this species. Intra-specific AT-rich DNA banding profiles were also obtained for isolates of A. cladogamus and these patterns corresponded well to the original hosts.

Two species of Melanochaeta and three species of Sporoschisma are reported from Thailand. Two connections of Sporoschisma and Melanochaeta are confirmed. Melanochaeta hemipsila has Sporoschisma saccardoi as its anamorph with a Chalara synanamorph in culture. Melanochaeta garethjonesii is described as a new species and linked with S. uniseptatum. S. nigroseptatum was recorded but no evidence of a Melanochaeta teleomorph state could be found.

Isolates of Cylindrocladium floridanum obtained from diseased roots of white and black spruce seedlings and surrounding soil in bareroot nurseries in Ontario, Minnesota and Wisconsin area were compared by mitochondrial DNA (mtDNA) and ribosomal DNA (rDNA) restriction fragment length polymorphisms (RFLPs), human minisatellite DNA fingerprints and polymerase chain reaction. Nucleotide sequences of rDNA ITS regions of selected isolates were also performed. Based on the nuclear DNA fingerprinting patterns, these isolates could be divided into three groups, namely S1, S2 and S3. Isolates of S1 and S2 showed the same mtDNA RFLPs which were slightly different from that of S3. Isolates representing each of the three groups showed an identical nucleotide sequence of rDNA ITS regions. These data suggest that all the isolates tested belong to the same species which may be divided into three biotypes or populations.

Collybia, as understood by Antonin & Noordeloos, comprises four species: C. racemosa, C. tuberosa, C. cirrhata and C. cookei. Collybia tuberosa, C. cirrhata and C. cookei are morphologically similar and are primarily distinguished from each other by the presence or absence and the colour of sclerotia. All four share a common and unique habitat. Phylogenetic reconstructions using DNA sequences of the ribosomal ITS1–5.8S–ITS2 support four distinct clades, each corresponding to a morphological species, with the C. tuberosa, C. cirrhata and C. cookei clades forming a larger group. Analyses of ribosomal Large Subunit DNA sequences confirmed that Collybia tuberosa, C. cirrhata and C. cookei formed a monophyletic group. In both analyses, the C. racemosa sequence was highly divergent from those of the other three species of the complex and we propose a separate genus name, Dendrocollybia, for this species. Simple diagnostic RFLP patterns were identified for the four species and were used to validate morphological designations and distributions.

Sclerotinia homoeocarpa is a common and severe fungal pathogen responsible for dollar spot disease on turfgrasses. The RAPD–PCR technique was used to identify genetic variation among 26 isolates of S. homoeocarpa. DNA polymorphisms were detected among different isolates using 15 oligonucleotide primers and a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA) for cluster analysis. Isolates from different geographical locations in the United States were separable into three distinct groups based on RAPD profiles and the cluster analysis. These three groups, however, were clearly more closely related to each other than to the isolate from Belize which was unique compared to the other S. homoeocarpa isolates examined. RAPD profiles suggest that transport of the pathogen from east coast to west coast locations in the U.S.A. may account for the recent identification of the disease in the Pacific Northwest. The results indicate that RAPD–PCR is a valuable method for differentiating isolates of S. homoeocarpa.

Types of asexual sporulation in 17 Claviceps species and the closely related Corallocytostroma ornicopreoides were revised in relation to the phylogeny of clavicipitaceous fungi. We observed: (1) enteroblastic conidiation from branched phialidic conidiophores typical of the genus (anamorph Sphacelia) in all species including Corallocytostroma; (2) widespread and often sequential formation of terminal holoblastic secondary conidia on tapering hyphae arising from sphacelial macroconidia; and (3) in addition to sphacelial conidiation, sympodial holoblastic conidiation of the Ephelis-type in cultures of C. zizaniae and in both the culture and sphacelial tissue of C. citrina. Secondary conidiation was not found in C. purpurea, C. citrina and C. sorghicola. During sphacelial fructification, most species produced macroconidia and microconidia. Only macroconidia formed in planta underwent secondary conidiation whereas microconidia did not germinate at all. In C. phalaridis, the formation of holoblastic 2–3 celled appendaged conidia was observed, similar to that of Aciculosporium and Neoclaviceps. In dendrograms based on ITS1-5.8S rDNA-ITS2 sequences, genera and species with appendaged conidia grouped on a highly supported clade with ancestral Corallocytostroma. The clade was placed inside a group of tropical species of Claviceps, without any relationship to Balansiae.

Several species of Typhula cause diseases of cereals and grasses at low temperatures. Taxonomic separation of T. phacorrhiza, T. incarnata and T. ishikariensis was investigated using RAPD, ITS-RFLP and nucleotide sequencing of the ITS region. Using 10 RAPD primers on genomic DNA or eight restriction enzymes on PCR-amplified ITS regions, banding patterns were generated for each of 19 isolates from five species or varieties. Polymorphisms were found between species and varieties, with fewer among isolates within species or varieties. RAPD analysis suggested that T. ishikariensis var. idahoensis may be distinct from the other varieties (var. ishikariensis and var. canadensis), although this was not supported by ITS-RFLP. To confirm relationships, ITS nucleotides were obtained from single isolates of each of the five Typhula taxa, and sequence alignments were subjected to distance and parsimony analysis. Although there were differences in branching between dendrograms derived from sequencing vs. RAPD or ITS-RFLP, sequence data supported the distinction of the three main Typhula species, with the T. ishikariensis varieties grouping together and showing differences at an infraspecific level. ITS sequence data also indicated the closer taxonomic affinity of Typhula species to the agaricoid Tricholoma sejunctum compared to the clavarioid Thelephora terrestris.

Ophiostoma setosum is described as a new species commonly associated with exposed sapwood of Tsuga heterophylla in western North America. It is characterized by the production of a typical ophiostomatoid teleomorph, a Pesotum anamorph with conspicuous, dark seta-like hyphae on the synnema stipes and apices, and a Sporothrix mycelial anamorph with solitary or short-catenate conidia. Mating experiments demonstrate that the species has two mating types controlled by two alleles at a single locus and is reproductively isolated from similar biological species of the O. piceae complex. Phylogenetic analysis of DNA sequences from the ITS2 and 5′ end of the large nuclear subunit (26S) of the ribosomal DNA supports the close relationship of O. setosum and O. piceae.

Isolates of Pithomyces chartarum obtained from pasture samples originating from Australia, Brazil, New Zealand and Uruguay were evaluated for their ability to produce sporidesmin. Only isolates that produced enough spores to confirm the presence or absence of sporidesmin were evaluated. Samples from the countries examined gave proportions of sporidesmin-producing isolates of: Australia, 67% of 207 isolates; Brazil, 2% of 51 isolates; New Zealand, 86% of 391 isolates and Uruguay, 28% of 182 isolates. Considerable variations in the ability to produce sporidesmin were found, not only between countries but between samples taken from the same country.

Using leaf disc techniques, nine isolates of the mycoparasite Sphaerellopsis filum were examined for pathogenicity to Melampsora epitea, the most important species of rust in renewable energy willow plantations. Five isolates were derived from Melampsora spp. on willows and poplar, one from Puccinia coronata on couch grass, two from Phragmidium violaceum on blackberry and one from Triphragmiopsis laricinum on larch. Two inoculation experiments were carried out. In the first, S. filum and rust were applied simultaneously on to leaf discs of Salix burjatica cv. Korso. In the second, leaf discs were inoculated with rust initially, then the resulting rust uredinial pustules were inoculated with S. filum. In both experiments, all S. filum isolates from Melampsora spp., and that from P. coronata, developed pycnidia on willow rust. No pycnidia were produced from the other three isolates. Among isolates which produced pycnidia, the frequency of rust pustules colonized by S. filum and suppression of rust spore production differed markedly. In the first experiment, 55–99% pustules were colonized and rust spore production was suppressed by 64–98%. In the second, the proportions were 33–97% and 53–73%, respectively. Suppression of rust sporulation was closely correlated with the frequency of rust pustules bearing S. filum pycnidia. The results suggest that S. filum is composed of pathogenically specialized populations differing widely in their virulence.