The effect of increasing culture age on cell surface hydrophobicity (CSH) and cell surface electrostatic charge (measured as zeta potential) of conidia from five isolates of Coniothyrium minitans representing three different morphological types was examined. Conidial CSH of three isolates (A2 960/1, CH1 and CH2) decreased with culture age, whereas CSH of two others (B 1300/2 and IMI 134523) remained high for the whole 42 day experimental period. In contrast, cell surface electrostatic charge decreased uniformly in conidia of all five isolates for the first 34 d and then rose slightly at 42 d. The variation in cell surface electrostatic charge (spectrum width) of the sampled conidia decreased with age for all five isolates. In all five isolates cell surface electrostatic charge of conidia became increasingly negative as the pH of the buffer used to suspend conidia was increased from pH 3·0 to 9·0. No relationship between colony morphology of C. minitans and conidial CSH and cell surface electrostatic charge was found.

The phylogeny of the Agyriales was investigated using nucleotide sequences of ITS1, 5.8S, and ITS2 rDNA. Sequences of these regions from 15 new sequences of 13 agyrialean fungi were aligned to those of four representatives of Ostropales used as outgroup. Six different alignments were analysed cladistically using maximum parsimony. The Agyriales were supported as a monophyletic group in the strict consensus tree and the synonymy of the Rimulariaceae and Saccomorphaceae was confirmed. The Schaereriaceae is also shown to belong to the Agyriaceae. The distribution of selected characters in the Agyriales is investigated and it is shown that the ascus type is variable within the group, while the ascoma ontogeny is invariant within the Agyriaceae. Trapelia as usually circumscribed is polyphyletic. The T. geochroa group is not closely related to Trapelia s. str. and the new genus Ainoa is described to accommodate T. geochroa and T. mooreana.

An extracellular protease designated BBP (Beauveria bassiana protease) was purified from a B. bassiana isolate and subsequently characterized. It was produced in the late stages of growth in a gelatin medium, and was purified using a combination of ion-exchange chromatography and preparative gel electrophoresis. Inhibition by phenylmethylsulphonyl fluoride and chymostatin indicates that BBP is a serine type with chymotrypsin activity. Substrate specificity indicates that the protease has elastase activity as well. The protease was stable at 25 °C and had an alkaline pH optimum (7.5–9.5). Another protease was purified from the same isolate that produced BBP, and was identified as Pr1 based on its isoelectric point, amino terminal sequence, substrate specificity and cuticle degrading ability. A comparison between the two showed that BBP had a lower isoelectric point (pI 7.5) than Pr1 (pI [ges ] 10) and was 0.5 kDa smaller. The proteases also exhibited differences in substrate specificity with Pr1 being most active against Suc-Ala-Ala-Pro-Phe-pNA, and BBP being most active against MeOSuc-Ala-Ala-Pro-Met-pNA. The two proteases had equal cuticle-degrading activity and both were expressed in cuticle media, although the timing of their expression differed.

A novel source of α-amylase has been identified in Pycnoporus sanguineus. Spores incubated in the submerged system using wheat bran as a carbon source showed α-amylase activity. Induction of α-amylase was also observed from mycelium incubated in submerged fermentation (LM4) using starch or wheat bran as the carbon source. The effect of the inhibitors on the production of enzyme by mycelium incubated in the presence of glucose, suggested the existence of catabolic repression. A comparative study showed that in a solid state fermentation system (SSM), even in the presence of high levels of glucose, the production of α-amylase was 4-fold greater than that in a submerged system. High levels of β-glucosidase and xylanase activities were also found in SSM after 72 h incubation. P. sanguineus produces at least three important hydrolytic enzymes using economical procedures.

The introduction of recombinant DNA technology in the field of mushroom research has resulted in the cloning and characterization of a large number of genes. In order to study the genetics of compost colonization of A. bisporus, genes encoding enzymes involved in utilization of this substrate have been isolated. In addition, a number of genes which are induced in fruit bodies during fruit body development have been cloned and they will provide more insight in the genetics of this economically important aspect of the life cycle. Other genes that were cloned encode proteins of basic biochemical routes. They provide knowledge on the importance and regulation of these routes in the life cycle of A. bisporus and add to knowledge on the general architecture of A. bisporus genes. Here we present an overview of the currently available biochemical and molecular data of A. bisporus and we discuss the importance of the available genes as genetic markers for breeding purposes.

The root systems of Carex flacca and C. pilulifera, growing in the Burren (western Ireland), were shown to be colonized by Cortinarius (Dermocybe) cinnamomeus. This basidiomycete formed ectomycorrhiza-like structures, which possessed a distinct fungal mantle (85–100 μm thick), hyphal infection in epidermal cells, rhizomorphs and extramatrical hyphae, but lacked a Hartig net. These ectomycorrhiza-like structures were formed on first-order lateral roots but were distinct, morphologically and anatomically, from dauciform roots. Structures typical of arbuscular mycorrhizas were never observed. The colonization of root systems of C. flacca by C. cinnamomeus was confirmed by PCR/RFLPs and DNA sequencing of the ITS region of rDNA.

The Nitschkiaceae has been placed in the Coronophorales or the Sordariales in recent years. Most recently it was accepted in the Coronophorales and placed in the Hypocreomycetidae based on sequence data from large subunit nrDNA. To confirm and corroborate the taxonomic placement and monophyly of the Coronophorales, additional taxa representing the diversity of the group were targeted for phylogenetic analysis using partial sequences of the large subunit nrDNA (LSU). Based on molecular data, the Coronophorales is found to be monophyletic and its placement in the Hypocreomycetidae is maintained. The order is a coherent group with morphologies that include superficial, often turbinate, often collabent ascomata that may or may not contain a quellkorper and asci that are often stipitate and at times polysporous. Three species with accepted Nitschkia names, together with Fracchiaea broomeiana and Acanthonitschkea argentinensis, comprise the paraphyletic nitschkiaceous complex. Two new families, Chaetosphaerellaceae and Scortechiniaceae fams nov., are described for the clades containing Chaetosphaerella and Crassochaeta and the taxa having a quellkorper (Euacanthe, Neofracchiaea and Scortechinia) respectively. The Bertiaceae is accepted for the clade containing Bertia species. Three new species are described: Bertia tropicalis, Lasiobertia portoricensis, and Nitschkia meniscoidea spp. nov.

The adhesion of conidia of Drechmeria coniospora to the nematode Panagrellus redivivus was reduced after treatment of the conidia with Pronase E, or the detergents sodium dodecyl sulphate (SDS) and dodecyltrimethylammonium bromide (DTAB) suggesting involvement of proteinaceous compounds in the adhesion process. In the TEM the thick extracellular pad covering the adhesive bud of the conidia was completely removed after treatment with Pronase E. After treatment with SDS or DTAB the proteinaceous compounds appeared to be dissolved leaving mainly carbohydrates in the pad as observed on OsO4 without and OsO4 with Ruthenium red-stained material, respectively. The detergent extracts after SDS and DTAB treatments contained nine and seven peptides, respectively, with molecular masses in the range from 6 to 80 kDa on SDS-PAGE gels, and five biotinylated peptides were found in each extract, after blotting to nitrocellulose membranes, indicating that these were surface proteins. None of the detergent extracts was able to reduce adhesion of the conidia after treatment of the nematodes. The detergent extracts contained protease- and phosphatase activity. The protease inhibitor, chymostatin, inhibited infection of nematodes and growth of the conidia, suggesting the involvement of chymotrypsin-like proteases in the infection process. On gelatin-containing substrate gel electrophoresis two proteases were clearly chymostatin sensitive.

Eighty-four isolates of Penicillium chrysogenum and related species were examined by isozyme analysis. Four main groups could be defined by cluster analysis; P. chrysogenum var. chrysogenum, P. flavigenum sp. nov., P. chrysogenum var. dipodomyis and P. nalgiovense. P. flavigenum is described as new species, and P. chrysogenum var. dipodomyis raised to species level as P. dipodomyis. The genetic distance between these groups was calculated by employing a formula that accounts for the differences in alleles and loci obtained by isozyme analysis of these fungi. The genetic distances and cluster analysis suggest that the groups should be considered as four separate species. The practical implication of the separation and the ecological characteristics of the four species are discussed.

The majority of microbes living on forest trees are still unnamed and our knowledge of their species richness is vague. This paper describes the fungal diversity of the above ground parts of a 61 year old Norway spruce tree lacking visible signs of damage or disease. The problem involved with identification of the fungi to named species was circumvented by classifying them into operational chemotaxonomic units (OCTUs) by using their combined fatty acid and sterol profiles (FAST-profiles). The variation of these units was chosen to correspond to a within-species-variation determined for several morphologically defined taxonomic species occurring on Norway spruce. Ninety-nine OCTUs were identified from 666 fungal isolates obtained. Bacteria were found only occasionally from the inner bark samples (three isolates) and from needles (five isolates). Models describing the accumulation of OCTUs against the number of samples taken were used for extrapolation of the total number of fungal OCTUs in the above ground parts of the tree. Our results suggest that an undamaged, apparently healthy Norway spruce, harbours in its above ground parts nearly two hundred fungal species. The majority were estimated to be needle epiphytes. At the large forest area scale, the species richness may be one order of magnitude higher.

All 39 marine species described so far from Spartina spp. are listed. Anthostomella torosa sp. nov. is described from culms of Juncus roemerianus in North Carolina, and A. spissitecta sp. nov. is described from leaf sheaths of saltmarsh Spartina alterniflora in the USA (Connecticut, Florida, North Carolina, Rhode Island) and from S. densiflora in Argentina (Buenos Aires). Both species occur close to the rhizome and are classified as obligate marine fungi. The new species are compared with similar taxa, and a table comprises characteristics of the nine known aquatic Anthostomella species.

The effect of Trichoderma atroviride, T. harzianum, T. longibrachiatum, Clonostachys rosea and Bacillus subtilis isolates applied to wheat seeds against Rhizoctonia cerealis disease of seedlings was investigated under controlled greenhouse conditions. Most Trichoderma isolates significantly reduced the incidence of disease compared with the infected control. Bacillus subtilis was also effective against sharp eyespot, although less active than Trichoderma spp. Interactions between the antagonistic microorganisms and the cereal pathogenic fungus in dual culture experiments on agar growth medium were also studied. Almost all tested antagonists showed competitive activity against R. cerealis: inhibition of its mycelial growth and hyphal interaction. The production of extracellular β-N-acetylhexosaminidase, chitin 1,4-β-chitobiosidase, glucan 1,3-β-glucosidase and protease activity by the tested microorganisms in the presence of cell walls of R. cerealis was then determined. All isolates showed glucosaminidase and chitobiosidase activity. They also produced glucosidase activity, except B. subtilis, whereas only C. rosea, B. subtilis and one isolate of T. harzianum showed detectable levels of protease activity.

Basidiomes of Amanita muscaria were collected from sites in three 30–36-yr-old Pinus radiata plantations in New South Wales, Australia. Following DNA extraction from basidiome stipes, inter-simple sequence repeat PCR using the degenerate primers 5′DHB(CGA)5 and 5′DDB(CCA)5, was used to identify the mycelial genotypes from which basidiomes arose. Each site was found to contain 8–10 A. muscaria genotypes, with six genotypes common to all three sites and one common to two sites. The presence of common genotypes at the three sites is taken to indicate that they were introduced as vegetative inocula when seedlings were planted and have persisted for < 36 yr.

Several varieties have been distinguished within the phytopathogenic pycnidial fungus Phoma exigua on the basis of cultural characteristics. These varieties have been stated to correlate with differences in ability to attack different host plants. Molecular discrimination of the varieties was investigated using amplified fragment length polymorphism (AFLP) and sequence analysis of the ITS region of the rDNA cluster on a set of 43 strains and 7 outgroup strains. The ITS sequences of the 43 different P. exigua strains were highly similar and revealed no subgroups within P. exigua while the AFLP fingerprint patterns of two primer combinations showed clear clustering of most varieties.

During surveys of woodlands in Taiwan a previously undescribed fungus Dryadomyces amasae gen. sp. nov., was found in the sapwood of dead angiosperm timbers in the gallery systems of the scolytine ambrosia beetle Amasa concitatus. The fungus grows predominantly in the immediate vicinity of the feeding beetle larvae and serves as a nutritional ambrosia fungus. The transmission of D. amasae to new breeding substrates is ensured by an oral mycetangium, a paired organ which was found in A. concitatus and A. aff. glaber near the mandibles and contained multiple cells of the fungus. Morphologically D. amasae resembles species of Ambrosiella. However, phylogenetic analysis based on partial nucSSU rDNA placed the fungus with certain species of the genus Ambrosiella within the Ophiostomatales, whereas the type species of Ambrosiella, A. xylebori, was assigned to the Microascales. A. xylebori, as well as A. ferruginea and A. hartigii, demonstrated phialidic conidial development, leading to an emendation of the description of the genus Ambrosiella. In contrast, Dryadomyces exhibited conidial development by apical, sympodial wall formation with prominent denticles bearing conidia. Other, previously described Ambrosiella species exhibited non-phialidic conidiogenesis, but lacked denticles on the conidiophores. Consequently, their classification needs further revision.

A new Claviceps species that causes ergot in sorghum and Sudan grass in Japan is described. The new species is distinct from C. sorghi and C. africana, from India and Africa, respectively, in the colour and texture of sclerotia, colour and morphology of stromata arising from the sclerotia, and size of ascospores and conidia. The pathogen infected only Sorghum in inoculation tests using 11 gramineous plants. Hyphal growth of C. sorghicola was 1 mm d−1 on PDA at 25 °C. Production of a different ergoline alkaloid in the sclerotia was suggested.

The sexual system of Heterobasidion insulare was elucidated and 40 specimens originating from the mainland of China, Taiwan, and Japan were studied with the aid of mating tests. Most of the material was also investigated with DNA fingerprinting using random amplified microsatellite (RAMS) and M13 minisatellite primers. Denaturing gradient gel electrophoresis (DGGE) and sequencing were applied for detecting variation in the internally transcribed spacer (ITS) region of the ribosomal DNA cluster. H. insulare proved to be a species complex with bipolar mating system. Clamp connections occurred in both heterokaryotic and homokaryotic mycelia. Three intersterility groups were found: the ‘T group’ included specimens from Taiwan and Guizhou Province (southern China), the ‘N group’ specimens from northern China and Japan, and the ‘Y group’ specimens from Yunnan (south-west China). The T group seems to be totally intersterile with the two other groups, and was grouped separately in neighbour joining analysis based on DNA fingerprints. As concluded from mating experiments, groups N and Y are partially intersterile, but they are able to produce hybrid heterokaryons, no group specific DNA markers were found, and they were grouped together in neighbour joining analysis. The identity of H. insulare sensu stricto was studied from the type collections. It seems that none of the above-presented taxa can be referred to it, and we propose the term H. insulare sensu lato to be used in future studies.

Strains of Heterobasidion annosum belonging to the S and P intersterility groups (IGs) were compared in respect of laccase activity on various substrates. Biomass production was significantly lower in P cultures than in S cultures. By contrast, laccase activity, measured in relation to growth rate, was significantly higher for the P strains tested, compared with a similar number of S strains, particularly in substrates, rich in carbohydrates as well as in inorganic and organic nitrogen. Using the Bradford assay method, P cultures were shown to contain significantly higher amounts of protein than S. The pH optimum of laccase for both IGs was 4·5 with guaiacol as substrate and 5·3 with syringaldazine. Isozyme patterns varied greatly, depending on strain, substrate and incubation time. In P cultures 4–5 bands were obtained, whereas S strain laccase was mostly separated into 2–3 isozyme bands. Only one was significantly different in position between the IGs. The results may help to explain why P strains are more aggressive than S strains as pathogens and wood decayers.

Mycelial interactions were studied between 28 isolates of Amylostereum areolatum and 15 of A. chailletii from wounded spruce in 10 sample plots in Sweden and Lithuania. Based on somatic incompatibility, nine vegetative compatibility groups (VCGs) of A. areolatum were detected. Five of these VCGs were isolated from two or more trees and included 24 (86%) of all isolates. In A. chailletii, 12 VCGs were found and three contained two isolates each. Grouping of isolates into VCGs was more pronounced in A. areolatum. Two of the A. areolatum VCGs had wide geographical distribution. VCG A2 was isolated from 14 different trees in two Swedish and four Lithuanian sample plots. VCG A1 was found in one plot in Sweden and in two in Lithuania. VCGs of A. chailletii had a local distribution and were confined to a single sample plot or to a forest stand. In A. areolatum occurrence of compatible pairings between isolates was uniform in all spatial scales of investigation: within a plot, within each country and among both countries. The proportion of somatically compatible A. chailletii isolates was significantly lower and decreased with geographic distance.

Rhodotorula mucilaginosa produces the siderophore rhodotorulic acid (RA) when grown in iron-limited conditions. R. mucilaginosa grew at rates between 0.10 and 0.19 h−1 in iron-restricted conditions, depending on the carbon source, and at 0.23 h−1 in iron-sufficient conditions. In bioreactors inoculated with iron-starved pre-cultures, initial specific growth rates in batch culture were dependent on the iron concentration. The critical dilution rate (Dcrit, at which steady state cultures cannot be sustained) in continuous cultures was also dependent on the iron concentration and was lower than μmax in batch culture. Sucrose was the best carbon source for RA production [287±11 μmol (g biomass)−1] and production could be further increased by supplementing the medium with the precursors acetate [460±13 μmol (g biomass)−1], ornithine [376±6 μmol (g biomass)−1], or both [539±15 μmol (g biomass)−1]. Citric acid was an effective suppresser of RA production. RA was produced in a growth rate dependent manner and was optimally produced at pH 6.5.