Interactions between springtails (Onychiurus armatus, O. tuberculatus, Folsomia candida) and plant pathogenic fungi (Gaeumannomyces graminis var. tritici, Fusarium culmorum, Bipolaris sorokiniana and Rhizoctonia cerealis) living in the same soil layer were investigated under laboratory conditions. The fungi are responsible for the foot and root disease complex of winter cereals and can cause serious reduction in yield. Since these species coexist in agricultural soils, feeding preference tests were performed in such a way that the springtails were allowed to interact simultaneously with all the tested fungi. O. armatus, O. tuberculatus and F. candida fed on the mycelia of G. graminis var. tritici, F. culmorum and R. cerealis: in the first hours of the trials individuals of all springtail species distributed themselves among the colonies, though moving from one to the other. Subsequently F. culmorum mycelium became the preferred food of all species tested; however, the other two fungi continued to be consumed. Mycelia of G. graminis var. tritici, F. culmorum and R. cerealis were shown to be an adequate food source for reproduction of Folsomia candida. Bipolaris sorokiniana mycelium had both repellent and lethal effects on F. candida and O. armatus, whereas this fungus was lethal, although not repellent, for O. tuberculatus. Long-term experiments on F. candida indicate that conidia of B. sorokiniana were eaten and proved to be a sufficient diet for reproduction.

Selected isolates of Phytophthora infestans from around England and Wales were fingerprinted using both RG57, a multi-locus RFLP probe, and Amplified Fragment Length Polymorphisms (AFLPs). The larger number of polymorphisms detectable with the AFLP method allowed resolution of several similar AFLP genotypes among isolates with identical RG57 fingerprints. However, some isolates with the same RG57 genotype had remarkably dissimilar AFLP genotypes, suggesting that there has been convergent evolution of some RG57 fingerprints. Also, some isolates with dissimilar RG57 fingerprints had similar or identical AFLP fingerprints. Both techniques distinguished isolates of mitochondrial DNA haplotype Ia from those of haplotype IIa. However, with AFLPs only, most of the isolates of A2 mating type were very similar and were distinguished from those of A1 mating type, suggesting that gene flow between A1 and A2 genotypes is limited and that sexual recombination is rare.

Several fungi associated with leaf spots and stem cankers of Proteaceae are reported. The new species Kabatiella proteae, Phloeosporella protearum, Septoria grandicipis, Stilbospora proteae and Trimmatostroma elginensis are described. Several new teleomorph-anamorph connections are reported, including Mycosphaerella stromatosa and Pseudocercospora stromatosa, Teratosphaeria microsporum and Trimmatostroma microsporum, and Lembosia proteae and Clasterosporium proteae. Additional species described are Lophiostoma fuckelii and Verrucisporota proteacearum which are also newly recorded associated with leaf spots of Proteaceae in South Africa.

The genetic variation of Duddingtonia flagrans, which has become a promising biocontrol agent of animal parasitic nematodes, was investigated in a worldwide collection of 22 isolates. We analysed the sequence variation in four nuclear genes, tubA (β-tubulin), CMD1 (calmodulin), EF1α (translation elongation factor 1α), and PII (extracellular serine protease). 1428 aligned base pairs (bp) were analysed from the four genes, including 709 bp of introns. In addition, the variations in three anonymous genomic regions comprising 1155 bp were examined. Three single nucleotide polymorphisms (SNPs) were detected in the seven loci, none of them in the protein encoding genes. The genetic variation was significantly higher in the nematode-trapping fungus Arthrobotrys oligospora, the closest evolutionary relative to D. flagrans. Analysis of 12 isolates of A. oligospora revealed 30 SNPs in tubA, CMD1, EF1α and PII. The genetic variation in the isolates of D. flagrans was further examined using AFLP analysis. Five primer combinations were used to detect 159 bands, of which 94 (59.1%) were polymorphic. A neighbour-joining tree based on the AFLP data showed no clear association between genotype and geographical origin. Furthermore, the AFLP data suggest that D. flagrans is mainly clonal and no recombination could be detected, not even within the same country. The low genetic variation in D. flagrans suggests that this fungus has recently diverged from a single progenitor. Based on estimations of mutation rates, it was calculated that this most recent common ancestor lived about 16000–23000 years ago.

A pair of primers specific for Neonectria galligena were designed from comparisons of the internal transcribed spacer (ITS) regions 1 and 2 of 32 isolates of diverse origins with sequences from 7 other nectriaceous species: ‘Nectria’ ditissima, N. coccinea, N. coccinea var. faginata, N. punicea, N. fuckeliana, N. cinnabarina, and N. radicicola. Comparisons were also made with sequences from several commonly encountered fungal endophyte species of apple wood, in particular Fusarium lateritium. Under stringent PCR conditions, primers Ch1 and Ch2 amplify a 412 bp fragment from N. galligena DNA nectriaceous but not from other nectriaceous species, fungal apple endophyte species, rosaceous, or beech host tissues. The identity of PCR products from total DNA extracts from woody tissues was confirmed by Southern hybridization. A 500 bp heterologous internal standard incorporating identical primer recognition sites was generated for use as an internal positive control for individual PCR assays to identify false negatives due to the presence of high levels of co-precipitated PCR inhibitory compounds. Further adaptation of this approach allowed for semi-quantitative competitive PCR enabling the determination of target fungal levels in infected lignified tissues.

The abilities of 12 ericoid mycorrhizal endophytes isolated from Woollsia pungens (Epacridaceae) and three Hymenoscyphus spp. isolates to grow on a range of amino acids as sole nitrogen sources were investigated in axenic liquid culture. The majority of endophytes grew to some extent on all amino acids tested, with most isolates from W. pungens showing a significant preference for histidine and/or alanine. Poorest growth of the endophytes from W. pungens was observed on lysine. In contrast, the Hymenoscyphus spp. isolates showed either no preference or a preference for lysine over histidine. Although some isolates produced some growth on the sulphur-containing amino acid cysteine, biomass production on this substrate by all isolates was significantly enhanced in the absence of inorganic sulphur.

The teleomorphic state of the ericoid mycorrhizal Hymenoscyphus ericae is known only from the type deposition. The production of both the teleomorph and anamorph by an isolate recovered from Ledum groenlandicum collected in an acidic peatland in Alberta, Canada, provided an opportunity to describe and illustrate the holomorph for a North American collection as a new record and as a supplement to the original diagnosis. It also provided further evidence that Hymenoscyphus ericae and Scytalidium vaccinii represent states of a single species, a hypothesis that previously had been tested using nuclear ribosomal DNA analysis. Appropriate cultural conditions and the use of molecular markers are advocated in order to facilitate the identification of mycorrhizal isolates which often remain sterile in pure culture.

The ability of the preferentially saprotrophic fungus Armillaria cepistipes and the pathogenic A. ostoyae to capture fresh stump resources was investigated in managed Alpine Norway spruce (Picea abies) forests where both species occur sympatrically. The incidence of Armillaria species and genets as rhizomorphs in the soil as well as external and internal (heart rot) stump colonisation were determined in three comparable 1 ha plots. The results indicate that A. cepistipes and A. ostoyae have a very similar strategy to capture fresh stump resources. Both species produce dense networks of rhizomorphs in the soil enveloping living trees. After the felling of the trees, the stumps are rapidly captured through spread of mycelial fans in the cambial zone. Our study suggests that A. ostoyae is slightly more competitive than A. cepistipes in primary resource capture.

The potential as biocontrol agents of the apple scab pathogen, Venturia inaequalis and the effect of environmental factors on growth, pycnidial production and spore germination of four isolates of Microsphaeropsis were examined on culture media and leaf discs. V. inaequalis ascospore production was reduced by 52, 77, 89 and 95% on leaf discs treated with the isolates IMI 294735, P176A, DAOM 198536 and P130A, respectively. For all isolates, optimum temperature for mycelial growth was 25 °C. More pycnidia were produced on culture media than on leaf discs. On culture media, optimum temperature was from 15 to 25 °, while on leaf discs it varied among isolates. Pycnidial production was inhibited by darkness and a minimum of 8 h light d−1 was required. Spores of isolates P130A and DAOM 198536 germinated at temperatures above 15 ° and at pH 4–5, as compared to 10 ° and pH 4–8 for isolates P176A and IMI 294735. The best candidates were P130A and DAOM 198536 and optimum conditions for growth, pycnidial production and spore germination were temperatures between 20 and 25 °, pH approx. 4 and light.

Micropeltopsis cannabis sp. nov. and Orbilia luteola (Roum.) comb. nov. are proposed. New Cannabis host associations include binucleate Rhizoctonia spp., Curvularia cymbopogonis, Sphaerotheca macularis, Glomus mosseae, and Pestalotiopsis sp. The geographic ranges of Pseudoperonspora cannabina, Septoria neocannabina and Fusarium graminearum are expanded.

Beauveria bassiana is a popular biocontrol agent used as ‘green’ pesticide in crop insect pest management. Chromosome number has been variously reported as five, six, seven and eight in this species. The range of chromosome number and the minimum chromosome number in this economically important fungus were assessed through telomere fingerprint analysis of a sample of 17 isolates from different and similar hosts and distant and same geographic origin. Genomic DNA digested with EcoRI, which has no cutting site in the telomere repeat sequence arrays was probed with a radioisotope-labelled (5′-TTAGGG-3′)8 oligonucleotide. The probe-hybridised regions appeared as discrete bands – each representing a telomere. The number of bands in each lane was counted and halved to arrive at the chromosome number of that isolate. The chromosome number varied from 5 to 10 in the different isolates. The telomere probe hybridised bands were also scored for presence or absence in a 0–1 matrix and a dendrogram based on similarities between the isolates was constructed using the NTSYS-pc ver. 2.02i software. The isolates showed very little similarity; the overall similarity was 14%. Only two isolates which were of diverse host and geographic origin showed 100% similarity. Isolates from the same epizootic that showed 43% similarity in their telomere fingerprints had 96% similarity in their RAPD (Random amplified polymorphic DNA) fingerprints with 10 primers. The genetic distances computed from any one DNA fingerprinting method thus do not reflect the true genetic similarities of the isolates. The frequency distribution pattern of the pair-wise similarities computed from telomere fingerprints hinted at the occurrence of recombination in this fungus. Telomere fingerprinting proved very useful in typing isolates since each of them was found to have a unique fingerprint. Isolates with the same chromosome number neither showed a distinct morphology or virulence character nor a close similarity in telomere or RAPD fingerprints to merit their subgrouping into a taxonomically relevant or practically useful unit.

Double-stranded (ds) RNA of various types was detected by electrophoresis in 23 of 25 isolates of Helicobasidium mompa. These dsRNAs varied in size from ca. 2 kbp to more than 10 kbp. dsRNAs from an isolate V1 had two distinct nucleotide sequences for putative RNA-dependent RNA polymerase (RDRP). Their complete sequences revealed that V1 dsRNA1 was 2247 bp in length, with a single ORF that encoded a 706-amino acid residue polypeptide with a predicted molecular mass of 82.6 kDa, and that V1 dsRNA3 was 1776 bp in length, with a single ORF that encoded a 538-amino acid residue polypeptide with a predicted molecular mass of 62.6 kDa. RDRP-conserved motifs were identified in both predicted amino acid sequences. Phylogenetic analysis indicated that V1 dsRNA1 was most closely related to Fusarium poae virus 1, while V1 dsRNA3 was most closely related to Helicobasidium mompa 70 virus. These results indicate coinfection of isolate V1 by two distinct partitiviruses.

275 isolates of Ophiostoma novo-ulmi, sampled across the southern Great Lakes region of North America from Wisconsin to Ohio in 1996, were analysed for vegetative compatibility (vc) types. Over 60% of the sample was a clonal vc component, comprising only two vc types (the AMSG and EUSG); the remainder of the sample was highly heterogeneous for vc types. Vc diversity was highest in Indiana and Michigan; close to where NAN O. novo-ulmi probably first appeared in North America. Each vc clone exhibited a uniform RAPD haplotype and a lower frequency of the A compared with the B mating type. This is consistent with the clones being mainly asexually spread, but with a potential for inbreeding, an unusual feature in a ‘genetic clone’. The heterogeneous component, however, exhibited diverse RAPD haplotypes and a near 1[ratio ]1 ratio of A[ratio ]B mating types, indicating that the mainly novel vc types of this component arise via sexual recombination. In Europe, dominant vc clones have been quickly replaced by novel vc types. In the Great Lakes region, however, vc clones appear to have survived for over 50 years despite a high potential for emergence of new vc types via sexual recombination. No differences were found between the clonal and heterogeneous components with regard to growth rate and pathogenicity, two important fitness parameters. Two other factors, a low level of selection imposed by deleterious d-factor viruses and a density dependent effect associated with vegetative incompatibility, may have favoured prolonged survival of the clones in North America.

Radiata pine (Pinus radiata) lumber from Australia, Chile, and New Zealand is imported into Korea where it represents one of the most important sources of timber. Blue stain of this timber is a serious problem for which an integrated control strategy is being developed. One of the fungi that has been isolated from stained radiata pine sapwood in Korea is a Leptographium sp. that has a distinct Sporothrix synanamorph. The aim of this study was to classify this fungus. Morphological comparisons showed that this fungus is distinct from all other species of Leptographium and especially L. elegans and L. francke-grosmanniae that also have Sporothrix synanamorphs. Comparisons of sequence data for the ITS2 and part of the 28S rDNA genes as well as the β-tubulin gene also confirmed that this fungus represents an undescribed taxon, described here as Leptographium bistatum sp. nov.

Tuberculina species are mitosporic parasites of rust fungi. Phylogenetically they belong to the Urediniomycetidae, therefore being closely related to their rust fungal hosts. We reveal by means of molecular analyses, ultrastructural and morphological features, observations in the field, and infection experiments that species of the genus Tuberculina and the violet root rot (Helicobasidium/Rhizoctonia crocorum) are stages of the life-cycle of one holomorph. This opens up new perspectives on parasitic life strategies as the resulting life-cycle is based on interkingdom host jumping between rusts and spermatophytes. In addition, we point at the consequences for any practical application dealing with Helicobasidium as an economically important plant pathogen and Tuberculina as a biological agent in rust control.

The diversity and frequency of leaf endophytic fungi occurring in naturally regenerating and planted stands of manuka were investigated at several sites. A restricted and characteristic range of species was isolated from all sites, although marked differences were noted between natural and planted stands. In natural stands a Phyllosticta species was the single dominant endophyte. This fungus appears to be host species specific, as it was not isolated from the closely related kanuka (Kunzea ericoides) in mixed, even-aged stands of manuka and kanuka. The frequency of the Phyllosticta sp. fluctuated greatly between individual trees across single stands. No obvious environmental or other gradients were recognized which could explain this patchy distribution. The Phyllosticta sp., along with the two next most common species in natural stands, Diploceras leptospermi and an unidentified coelomycete, were absent from most of the planted stands sampled, and at very low frequencies when present. Although several of the planted stands were close to natural stands and had been established for several years, the difference between them has been maintained. The most common endophytes in planted stands were Botryosphaeria and Alternaria spp. These species, found at low levels in natural stands, are likely to be ‘opportunistic’ endophytes. They are probably also characteristic of the epiphytic mycota, invading only the living leaves of plants under stress. The difference in leaf endophyte communities between planted and natural stands of manuka may be an indication of wider differences in the microbial communities of the respective stands, and could prove significant for site restoration projects. Such projects using nursery-raised manuka plants as a pioneer species may be establishing stands lacking much of the natural diversity of manuka communities for extended periods following establishment.

Sarcodon imbricatus has long been used to extract blue and greenish pigments for wool dyeing. We found that fruit bodies growing with Pinus sylvestris seemed to be superior for dyeing compared to fruit bodies growing with Picea abies, and macroscopical differences between the forms indicated that they are different taxa. By studying sequences of rDNA ITS and macroscopical characters, two species were recognized. Sarcodon imbricatus grows in association with Picea, and S. squamosus with Pinus. The latter species, described by Schaeffer in 1774, has been lumped with S. imbricatus during the past 50 years, creating great confusion among wool dyers.

Colletotrichum acutatum is cosmopolitan and causes diseases in many crops including Hevea brasiliensis. It sporulated freely on PDA at 10–40 °C with peaks at around 15° and 25°. Self-inhibition of spore germination occurred at concentrations above 8×106 spores ml−1. Ultraviolet radiation (especially 254 nm) inactivated the spores. Spore germination was around 90% between 15–35°. Free water promoted spore germination but was not essential; high relative humidity (95%) was sufficient. Spores could also withstand temperatures up to 35°. It is concluded that the most favourable conditions for the spread of C. acutatum are prevalent in the major rubber growing areas in Sri Lanka throughout the monsoons.

The relation between the pattern of fumonisin production and the mating population and mating type of Fusarium isolates from maize in Argentina at different maturity stages has been evaluated. Fifty-one isolates of Fusarium species belonging to the Liseola section were identified to mating population and tested for their ability to produce fumonisins (FB1, FB2 and FB3). Only mating populations associated with maize, A (Fusarium moniliforme, 23 isolates), D (F. proliferatum, 24 isolates) and E (F. subglutinans, 4 isolates) were found. All but two isolates of populations A and D produced, when grown on maize substrate, high levels of fumonisins ranging from 0·01 to 3·99 mg g−1 (mean 2·00 mg g−1 and 1·69 mg g−1, respectively), whereas isolates of population E yielded less than 0·02 mg g−1 (mean 0·01 mg g−1). Five isolates of F. proliferatum, all belonging to mating type D−, produced more FB2, than FB1, which is consistent with finding a relatively large amount of FB2 (0·01 mg g−1; FB2/FB1=2·27) in one of the maize samples from which these isolates were obtained. Amounts of FB3 were similar to FB2 in cultures of mating population A (mean FB3/FB2=0·89), but much lower than FB2 in cultures of mating population D (mean FB3/FB2=0·21).

The edible mushroom, Agaricus bisporus, when cultivated commercially derives its nitrogen from the proteins present in compost, much of which is firmly bound to the lignin-humic complex. The proteinases involved in mobilization of nitrogen were studied in both the filtrates of liquid culture and mushroom compost. Proteinase activities were detected in liquid culture filtrates with a range of substrate specificities. One activity with chymotrypsin-like specificity was found by Western blotting to be the 27 kDa serine proteinase previously purified from senescent sporophores. Serine proteinase activity and level in the filtrates rapidly increased during mycelial growth on liquid medium containing humic fraction (extracted from mushroom compost) as sole nitrogen source. Activity and level remained low in filtrates of cultures grown on glutamate or casein. Non-denaturing gel analysis revealed the serine proteinase to be the most prominent proteolytic activity in compost filtrates. Regulation of the activity in compost was indicated by a doubling during the early stages of sporophore growth.