Interrelations of fungal mycelium with other soil biota are of paramount importance in forestry and soil ecology. Here we present the results of statistical analysis of a comprehensive data set collected in the first (and the only) British fungus sanctuary over a period of four months. The variables studied included a number of soil properties, bacteria, protozoan flagellates, ciliates and amoebae, microbial and plant feeding nematodes, various microarthropods, and two fungal biomarkers – glomalin and ergosterol. One way ANOVA showed that the dynamics of the microbiota studied was influenced by seasonal changes. Superimposed on these changes, however, was variability due to biological interactions and habitat characteristics. Two fungal biomarkers, ergosterol and glomalin, were differently influenced by other biota and abiotic variables. The results indicate that the dynamics of soil fungi is influenced not only by soil microarthropods, but also by those found in forest litter. The overall outcome, therefore, is likely to be very complex and will depend upon specific conditions of any particular ecosystem.

A survey of Penicillium albocoremium was undertaken to identify potential taxonomic metabolite markers. One major and four minor metabolites were consistently produced by the 19 strains surveyed on three different media. Following purification and spectral studies, the metabolites were identified as the known protein farnesyl transferase inhibitors andrastin A (1) and barceloneic acid A (2) along with barceloneic acid B (3), barceloneic lactone (4), and methyl barceloneate (5). These compounds are significant taxonomic markers for P. albocoremium; moreover this is the first report of a methyl ester of a barceloneic acid being produced as a secondary metabolite. Tissue extracts created following pathogenicity trials involving P. albocoremium and Allium cepa confirmed the production of these five metabolites in planta. Barceloneic acid B was found to be biologically active against a P388 murine leukemia cell line.

Aquaphila albicans gen. and sp. nov. from submerged wood in the tropics is described and illustrated. It produces hyaline, multiseptate, fusoid or falcate conidia resembling the macroconidia of Fusarium species, but differs by its conidiogenesis and the aquatic habitat. The conidiophores of A. albicans are septate, hyaline, sympodially proliferating with conidiogenous denticles. Conidial development in A. albicans has been studied in culture and results are presented. The genus is compared with similar genera.

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.

The patterns of conidial discharge of Erynia neoaphidis were measured from three species of aphids: nettle aphid (Microlophium carnosum), grain aphid (Sitobion avenae) and pea aphid (Acyrthosiphon pisum). The effects of release height, morph of aphid and temperature on the horizontal and vertical discharge of conidia were studied. Numbers of conidia deposited in the dorsal and lateral directions were distributed with distance in truncated bell shaped patterns. Discharge distances ranged from 2 to 11 mm and half the conidia caught travelled further than 5 mm. There was little difference in the horizontal dispersal patterns of conidia when ejected from cadavers of different morphs of the same species, or from different species. The weight of infected aphids showed little correlation with discharge patterns. Values of maximum discharge distance, Dm, were generally between 6 and 9 mm. Temperature had a significant effect on conidia discharged from the dorsal surface of apterous A. pisum cadavers. Dm was greater at 18 °C than at 10 or 25 °. Vertical discharge distances for conidia released from A. pisum cadavers ranged between about 2 and 8 mm. The maximum height to which conidia were projected vertically was not affected by temperature. However, the average height tended to be greater at 18 °. Half of the conidia reached heights of about 3.5 mm while the maximum heights reached were about 8 mm. The initial speeds of conidia were estimated from measured discharge distances. The results suggest that conidia of E. neoaphidis may be ejected at speeds of about 8 ms−1.

Apple fruits and grapevine leaves were inoculated with four wild-type strains of Botrytis cinerea. Two types of symptoms were observed on apple, firm or soft rot. Extracts from infected tissues were analysed for polygalacturonase (PG) and pectin methylesterase (PME) isozyme profiles. They showed that different specific PG isozymes occur in grapevine leaves and in soft rot on apple fruit. PG and PME profiles were also studied in genetic transformants altered in their colonization ability and/or the type of rot that they caused on apple fruit. No PG activity was found in apples infected by firm-rotting wild-type strains or transformants, whereas two neutral PG isozymes (pIs 7.3 and 7.6) were expressed in soft-rotting strains. No association was found between the production of these PG isozymes and the colonization level. The PME patterns were not correlated with the rotting type nor the colonization level. In vitro, firm-rotting strains and transformants showed no PG activity in glucose medium whereas soft-rotting ones produced several PG isozymes. In polygalacturonic acid cultures, all strains and transformants produced a single, common acidic isozyme. Unlike PG, no variation in the PME isozyme patterns in vitro could be correlated with the culture substrate on apple or the rotting type. The variation in pectinase profiles between in vitro and in planta contexts and the role of PGs in B. cinerea pathogenicity on apple fruit is discussed. We suggest that neutral PG isozymes are involved in the soft-rotting type on apple fruit.

A thermostable extracellular glucoamylase from Thermomyces lanuginosus in static culture was purified to SDS–PAGE homogeneity by fractional ammonium sulphate precipitation, ion-exchange chromatography on DEAE–Toyopearl, Butyl–Toyopearl hydrophobic interaction chromatography, gel filtration on Sephacryl S-300 and ion-exchange chromatography on FPLC MonoQ. The molecular weight of the enzyme, consisting of a single polypeptide, was estimated to be 72000 by SDS–PAGE. The glucoamylase exhibited maximal activities at pH 5·0. The optimum temperature for the activity was 70°C. The enzyme was thermostable at 60° with half-lives at 70° of 20 min and 80° of about 6 min. The glucoamylase was a glycoprotein with 11·4% carbohydrate content. The enzyme hydrolysed soluble starch, amylose, amylopectin, dextrin, glycogen, maltotroise and maltose. Starch was the best substrate.

Cronartium flaccidum and Peridermium pini are rust fungi occurring on two-needle hard pines. According to previous molecular and morphological analyses, they are very closely related despite differences in their life-cycles. In this study we showed that although a low level of genetic differentiation occurs among populations of both P. pini and C. flaccidum, there is no overall differentiation between the two rusts, and in this respect they resemble a single taxon. We also observed evidence for linkage disequilibrium occurring between different alleles of separate loci of Peridermium pini suggesting that its population structure would be clonal. This suggests that strains of P. pini would have originated as asexual or self-fertilizing host range mutants of C. flaccidum.

Changes in pre- and post-fire biomass of hypogeous fungal sporocarps remaining at locations where an endangered mycophagous marsupial, the northern bettong (Bettongia tropica), had foraged, were assessed in fire-prone sclerophyll forest in northeastern Australia. Low to medium intensity experimental fires were set during the late dry season in 1995 and 1996 and post-foraging biomass of sporocarps (expressed as biomass of sporocarps remaining at recent B. tropica diggings) was measured at unburnt and burnt sites at approximately six-week intervals for a period of 14 months. Post-foraging biomass was significantly higher at burnt sites immediately following fire compared with control sites, solely due to increased biomass of hypogeous species belonging to the family Mesophelliaceae. Several months after fire, post-foraging biomass was significantly higher on unburnt sites compared with very low biomass on burnt sites. Twelve months after fire, the biomass on burnt and unburnt sites was not significantly different, having returned to biomass observed pre-fire. All evidence points toward mesophellioid fungi being greatly more available to bettongs on recently burnt ground, but fire may make several other sporocarp taxa considerably less available several months following fire.

The Claviceps purpurea population colonising British Spartina salt marsh stands is characterised by unusually long cylindrical conidia (average 9·5–12 μm) and sclerotia floating on the water surface. RAPD, AFLP and rDNA comparison defined these isolates as the third genetically distinct homogenous population (G3) of C. purpurea. The same morphological and genetical markers were found also in S. alterniflora isolates from Spartina from the USA. All G3 isolates belonged to a chemotype producing ergocristine and ergocryptine. In phylogenetic trees based on rDNA and AFLP, a G1 population from fields and meadows appeared as the sister clade to the one formed by G3 (Spartina) and G2 (wet and shady habitats), both with floating sclerotia and elongated conidia. British stands of S. anglica were probably colonised by isolates introduced from America, rather than by isolates from species of neighbouring biotopes.

Whole genome sequencing of several filamentous ascomycetes is complete or in progress; these species, such as Aspergillus nidulans, are relatives of Saccharomyces cerevisiae. However, their genomes are much larger and their gene structure more complex, with genes often containing multiple introns. Automated annotation programs can quickly identify open reading frames for hypothetical genes, many of which will be conserved across large evolutionary distances, but further information is required to confirm functional assignments. We describe a comparative and functional genomics approach using sequence alignments and gene expression data to predict the function of Aspergillus nidulans genes. By highlighting examples of discrepancies between the automated genome annotation and cDNA or EST sequencing, we demonstrate that the greater complexity of gene structure in filamentous fungi demands independent data on gene expression and the gene sequence be used to make confident functional assignments.

To identify Stagonospora avenae genetically, 53 isolates from Europe and North America were assessed by RFLP analysis and internal transcribed spacer (ITS) sequence polymorphisms in comparison with isolates of S. nodorum, S. arenaria and Septoria tritici. Our results also provided insight on possible phylogenetic divergence within the species currently identified by characteristic conidial morphology as S. avenae. Isolates from Europe, Canada, Minnesota and North Dakota showed little genetic variation. In RFLP and ITS sequence analyses, S. avenae isolates from New York and ATCC 12277 (origin unknown) had a closer relationship with barley-biotype S. nodorum, and S. avenae f. sp. triticea isolates of ATCC 26370 and ATCC 26377 from Minnesota were closely associated with wheat-biotype S. nodorum. Enzyme restrictions of ITS PCR-amplified products could facilitate the identification of Stagonospora on cereals.

Cellulose-acetate electrophoresis (CAE) was used to investigate isozyme polymorphisms of 34 Group 1 and Group 2 isolates of Fusarium graminearum. Of the 26 enzyme systems screened, alkaline phosphatase, NADP-dependent glutamate dehydrogenase, peptidase D and phosphoglucomutase were found to be suitable for distinguishing members of Group 1 from members of Group 2. A great deal of similarity in the isozyme patterns among the isolates of a same group, independently of geographic origin, indicated that isolates within a given group are descendant from a same ancestral population. Significant differences in the isozyme patterns between the isolates of Group 1 and Group 2 indicated, however, that the two groups were of different ancestry. Group 1 isolates have since been named F. pseudograminearum. Due to the short run time, minimal need of equipment, small sample volume and staining requirements, CAE generally provides a rapid and accurate method for isozyme analysis of Fusarium species.

A molecular study of the mycorrhizal symbiosis between barley and Glomus intraradices used differential display PCR and a synchronous colonization method to identify genes that are differentially expressed in symbiosis. Several PCR products were consistently differentially amplified. PCR amplification of genomic DNA from either G. intraradices or barley as templates showed that three such products were encoded by G. intraradices. Sequence analysis of the deduced amino acid sequences of the fungal fragments, following extension by 3′-RACE, revealed similarities to proteins from higher eukaryotes. One (GINMYC1) shows similarity to TRIP15, a human protein that interacts in a hormone-dependent manner with the thyroid receptor. A second (GINMYC2) is similar to O-linked N-acetylglucosamine transferases from vertebrates, and the third (GINHB1) contains a putative leucine zipper and a homeodomain which indicates that it binds DNA and may act as a transcriptional regulator. Fragments of the expected sizes were amplified by RT–PCR from mRNA of mycorrhizal barley roots for all three fungal cDNAs, which indicates that the corresponding genes are expressed during intraradical growth of G. intraradices. The results provide a promising insight to fungal gene expression early in formation of this compatible and mutualistic symbiosis.

On water agarose conidial germination of Ampelomyces quisqualis was greatly enhanced in the presence of powdery mildew conidia. All seven species of powdery mildews tested, representing five genera, were effective in increasing germination of the mycoparasite. Aqueous extract of conidia of Oidium euonymi-japonicae was also stimulatory to the mycoparasite. Germination of conidia of A. quisqualis on water agarose decreased as conidial concentration increased. Germination of the mycoparasite's conidia at a low concentration on water agarose was greatly decreased when they were separated by a polycarbonate membrane from conidia of the same species at a high concentration and incubated. After being incubated with conidia of the mycoparasite at a high concentration, water agarose became inhibitory to germination of conidia of the same species at a low concentration, suggesting the presence of a diffusible self-inhibitor. Water agarose is essentially free from nutrients which may support growth of contaminants that interfere with the effect of stimulators or inhibitors.

Variation in Gremmeniella abietina var. abietina was studied in three stands of Scots pine in northern Finland and in the Kola Peninsula. Eighty-four isolates of large- and small-tree types of G. abietina var. abietina (LTT and STT, respectively) were identified on the basis of tentative characteristics (spore morphology, disease type and host size), fatty acid and sterol profiles (FAST), and random amplified microsatellite technique (RAMS). Both LTT and STT occurred in all three stands. In general, the classifications obtained using the three methods agreed with one another, although a few contradicting results were observed. Variation in fatty acids and sterols in G. abietina var. abietina was rather low, although the amounts of some individual extractives showed statistically significant differences between the stands. All pathogenic and asymptotic G. abietina var. abietina isolates originating from branches located at heights above the annual snow cover were identified as LTT based on RAMS, but some were grouped to STT according to their FAST profiles. Both STT and LTT were detected among the isolates obtained from seedlings according to both FAST and RAMS. In addition, in two cases RAMS markers thought to be STT- or LTT-specific were found in the same isolate. The results presented here suggest that LTT of G. abietina var. abietina caused the devastating epidemics on pines in the first-thinning stage or middle age similar to pines in this study reported in northern Finland and in the Kola Peninsula during the 1980s.

Sordaria brevicollis is a filamentous ascomycete previously considered to be exclusively heterothallic. Here we show that S. brevicollis can exhibit homothallism, and describe the genetic and environmental factors favouring the process. Both mating types can produce perithecia in unmated cultures, but only in the uncrossed perithecia of mtA are ascospores formed (in <30% of the fruitbodies). In most cases, only one or two asci are produced in any uncrossed mtA perithecium, although as many as 13 have been observed. The ascospores show no segregation for mating type; all are mtA. Heterokaryon analysis established that meiosis and recombination occurred during ascospore production in both homothallic and heterothallic development. Homothallic fruiting is inhibited both by crossing with the opposite mating type, and by light.

The genus Leptographium was described in 1927 and currently includes 48 species, with L. lundbergii as the type species. In recent years, the taxonomic status of L. lundbergii has not been uniformly agreed upon and it has been the topic of considerable debate. The problem was compounded by the absence of a type specimen, and the species was epitypified at a later stage. Unfortunately, the whereabouts of the epitype is now unknown. In 1983, Wingfield & Marasas described L. truncatum, which is morphologically similar to L. lundbergii. Based on DNA comparisons and similarities in their morphology, this fungus was reduced to synonymy with L. lundbergii. The loss of the type specimen as well as variation in the morphology of strains identified as L. lundbergii prompted us to re-examine the taxonomic status of this species. A number of strains from various geographic areas were studied. These include a strain of L. lundbergii deposited at CBS by Melin in 1929 (CBS 352.29) as well as the ex-type strain of L. truncatum. The strains were compared based on morphology and comparison of multiple gene sequences. Three genes or genic regions, ITS2 and part of the 28S gene, partial β-tubulin and partial elongation factor 1-α were compared. Strains currently identified as L. lundbergii, represented a complex of species. Strains initially described as L. truncatum clustered separately from other L. lundbergii strains, could be distinguished morphologically and should be treated as a distinct taxon. L. lundbergii is provided with a new and expanded description based on a neotype designated for it. A third group was also identified as separate from the main L. lundbergii clade and had a distinct Hyalorhinocladiella-type anamorph, described here as H. pinicola sp. nov.

Species delimitation is still controversial in the Xerocomus chrysenteron complex. We have therefore established comprehensible and reliable species concepts based on statistical evaluation of morphological and ecological characters. We examined many collections from different geographical regions and different developmental stages within collections. Quantitative micromorphological characters (basidiospores, pileipellis end cells) were measured in statistically relevant numbers. The same material was used to generate 24 rDNA-LSU sequences, and the results of phylogenetic analyses clearly confirmed our species concepts: spore size and ornamentation, length of the pileipellis end cells and ‘pruinatus-hyphae’ are most valuable characters for the delimitation of species in this complex. Molecular data demonstrated that the X. chrysenteron complex is a monophyletic group. All the examined species (X. chrysenteron, X. cisalpinus, X. pruinatus, X. ripariellus, X. dryophilus, X. fennicus, X. porosporus, and X. rubellus) represent independent lineages. The faintly striate spores, a key character characterising species of section Striatulispori, probably evolved independently. In addition, the ‘pruinatus-hyphae’ have multiple origins, and truncate spore apices are derived at least twice. Xerocomus cisalpinus sp. nov. is characterised by striate spores, the presence of ‘pruinatus-hyphae’ and a pileipellis strongly reminiscent of X. chrysenteron. For reasons of discussion, microscopical data are presented on Boletellus episcopalis for the first time. Xerocomus fennicus (Boletellus) comb. nov. is proposed. We provide descriptions to all included taxa. Our results once more demonstrate that reliably identified and characterised voucher collections are the basic requirement for meaningful phylogenetic studies.

The suitability of random amplified polymorphic DNA for identification of Pythium aphanidermatum was investigated. Three oligoprimers were selected after testing three isolates of P. aphanidermatum and one isolate of P. ultimum with a total of 40 primers. The selected 10-mer primers were used with 20 isolates of P. aphanidermatum, four isolates of P. deliense, two isolates of P. ultimum, two isolates of P. irregulare and one isolate of P. paroecandrum. Most of the P. aphanidermatum isolates (13 of 20), were obtained from samples of Cucumis sativus, or water from cucumber greenhouses. The three selected primers gave identical fingerprints for 18 of the 20 P. aphanidermatum isolates, including all the isolates from cucumbers. Two of the primers gave fingerprints that could be used to differentiate between isolates of the Pythium species studied. The banding pattern of P. aphanidermatum given by the third primer could not easily be distinguished from the fingerprint of P. deliense. However, when used in conjunction with the other two primers, the third primer can be used to verify the identity of P. aphanidermatum.