An undescribed species of the heterobasidiomycetous genus Chionosphaera is carried by the bark beetles Dryocoetus autographus, Hylurgops palliatus, Ips acuminatus, Ips sexdentatus, Ips typographus, Orthotomicus laricis, Pityogenes chalcographus, Pityokteines spinidens, and Polygraphus poligraphus infesting conifers in Europe (Abies alba, Larix decidua, Picea abies, and Pinus sylvestris). Chionosphaera cuniculicola sp. nov. differs morphologically from C. apobasidialis in the more slender basidiospores. The segregation of the two species is supported by comparison of partial sequences of the large subunit of the ribosomal gene. Isotype material of Chionosphaera lichenicola was re-examined. In contrast to C. cuniculicola and C. apobasidialis, C. lichenicola exhibits clamps at the septa. Fibulostilbum phylacicola is considered as a clamp-bearing species of Chionosphaera.

The genetic structure of five Fennoscandian populations of the threatened wood-decay fungus Fomitopsis rosea (Basidiomycota) was investigated using codominant PCR–RFLP, allele specific amplification (ASA) markers, inter simple sequence repeat (ISSR) markers and mating studies. Sequence analyses of a subset of single spore isolates revealed sequence variation in four target sequences; internal transcribed spacer (ITS) and intergenic spacer (IGS1) of the nuclear ribosomal DNA, the translation elongation factor 1 alpha (efa) gene and the super oxide dismutase (sod) gene. No sequence variation was found in amplified portions of the mitochondrial large and small rRNA genes. Genotype distributions were mostly (90%) in accordance with Hardy–Weinberg expectations, and the nrDNA markers (ITS/IGS1), efa and sod were in most cases (87%) in linkage equilibrium, indicating an outcrossing reproductive mode, panmictic conditions and large population sizes of the fungus. Mating tests confirmed that F. rosea exhibits an outcrossing bipolar heterothallic mating system. Mating allele richness was high in two investigated populations. Phylogenetic analyses of ITS and IGS1 sequences from the five geographic populations revealed some geographic sub-structuring of the ITS sequences, but no sub-structuring of IGS1. The nrDNA (ITS/IGS1), efa and sod markers gave a low overall FST (0.013). The ISSR markers gave no clustering of the populations in UPGMA, and the between-population variance component was very low in AMOVA (0.4%), indicating a high level of gene flow.

Selenate-resistant (SelR) mutants of Botrytis cinerea were selected by plating conidia or mycelial plugs onto minimal medium amended with selenate and taurine. Ultraviolet irradiation of conidia to give 5% survival increased mutant yield twenty-fold. Mutants could be divided into three classes based on growth in the presence of selenate or chromate and on response to taurine in minimal media. Nitrate non-utilizing (Nit) mutants, generated as spontaneous sectors on minimal media amended with chlorate, behaved as nit1 mutants in growth tests (i.e. putatively defective in nitrate reductase apoenzyme). When nit1 mutants were paired on medium with nitrate as sole nitrogen source some pairings complemented, behaviour attributed to intragenic complementation. Selected crosses of SelR and nit1 mutants with wild-type strains gave 1[ratio ]1 segregation of both phenotypes and no evidence of linkage to either Mbc1 (benzimidazole resistance) or Daf1 (dicarboximide resistance) markers; loose linkage was confirmed between Mbc1 and Daf1. Strains showing the SelR phenotype may result from mutations in different genes; the genotypic symbol Sel 1 is allocated to one of these mutations. Four-point crosses involving the markers at Sel 1, nit1, Mbc1 and Daf1 generated progeny with all 16 genotypes. Both Sel1R and nit1 mutants were stable following subculture and retained pathogenicity in a bean seedling assay. Complementation was demonstrated between SelR and nit1 mutants selected from the same parent.

Extracts from entomopathogenic fungi of the genus Hypocrella (7 species) and its anamorph Aschersonia (11 species) were screened for cytotoxicity to Sf9 and C6/36 insect cells and L929, BHK(21)C13 and HepG2 mammalian cells. Cytotoxic extracts to insect cells (ID50's[les ]10 μg ml−1) but not to mammalian cells (ID50's[ges ]10 μg ml−1) conformed to the criteria of the project and were considered ‘lead’ extracts for further investigation. ‘Leads’ were found in two of the Hypocrella species: H. discoidea, and H. tamurai and in three of the Aschersonia species: A. samoensis, A. badia, and A. tamurai. Bioassay-guided fractionation of the cell extract of the fungus A. samoensis BCC 1393 led to the identification of two known anthraquinone dimers, (+)rugulosin (1) and skyrin (2) which showed selective toxicity towards insect cells. (+)Rugulosin (1) and skyrin (2) exhibited strong cytotoxic activity against the insect cell line Sf9 with respective ID50 values of 1.2 and 9.6 μg ml−1, but showed weak activity toward mammalian cells. This first report of (+)rugulosin (1) and skyrin (2) in A. samoensis is confirmed and demonstrated in another four strains of A. samoensis isolated in Thailand. The preferential cytotoxicity against Sf9 insect cells gives evidence that these insect-pathogenic fungi of the Hypocrella/Aschersonia group might be useful as an agent for pest control.

As one of the endangered breed of mycologists to have maintained an interest in oomycetes, I am compelled to defend the rationale in continuing to refer to these micro-organisms as fungi. This paper is stimulated by recent communications in which my mycological colleagues have, with apparent relish, pointed to what they perceive as inaccurate references to oomycetes as fungi. David Moore (1997) provides a prime example in his review of The Growing Fungus edited by N. A. R. Gow & G. M. Gadd (1995). Moore criticizes the editors for allowing so many pages of their book to be devoted to these micro-organisms, when ‘…Neurospora is probably more closely related to a cow's nose than it is to Saprolegnia’. While a phylogenetic chasm separates the oomycetes from other fungi, I shall argue that it is impractical to restrict the usage of the term fungus to those micro-organisms that qualify as members of the Phylum Fungi (Alexopoulos, Mims & Blackwell, 1996, provide a contemporary overview of fungal systematics). This argument should have been settled by a series of clear-sighted articles published after molecular genetic analyses confirmed the polyphyletic nature of the fungi (Christensen, 1990; Bruns, White & Taylor, 1991; Hawksworth, 1991; Barr, 1992). For example, Donald Barr (1992) suggested that oomycetes are fungi in the colloquial sense of the term and that it is the colloquial sense of the term that will continue to be most meaningful to mycologists. But since the message of these articles was apparently missed by many mycologists, I hope that this note will clarify a practical definition of the fungi that includes the oomycetes but which does not conflict with a natural classification of the Phylum Fungi that excludes them.

Restriction fragment length polymorphisms (RFLPs) were identified among total DNA from clonal lines of Uncinula necator when cloned sequences of total U. necator DNA were used as probes. Four probes, pUnP14, pUnP27, pUnE21 and pUnE4, hybridized to multiple-copy sequences and, with the exception of pUnE4, detected genetic variation among clonal lines of U. necator. Clones pUnP14, pUnP27 and pUnE4 produced banding patterns that were stable for DNA extracted from different asexual generations of U. necator clonal lines over at least 15 months. In addition, clones were evaluated for species specificity. Clones pUnP27 and pUnE4 detected only U. necator sequences in total DNA from infected grapevine leaves. Clones pUnP14 and pUnE4 did not hybridize to total DNA from a range of fungi.

Genetic diversity in a sample of the Australian U. necator population was investigated; 15 genotypes were identified among 29 U. necator clonal lines examined. Genetic variation was detected in samples collected within micro-geographical areas, for example, different genotypes representing both mating types of U. necator were detected on a single plant of Vitis amurensis. RFLP analysis of the banding patterns produced using pUnP14, pUnP27 and pUnE21, identified two broad genetic groups, designated A and B. Analysis of DNA fragment patterns obtained using the polymerase chain reaction (PCR) and the plant intron splice junction (ISJ) primer R1 also supported the allocation of U. necator clonal lines into groups A and B.

Sequences of the intergenic spacer between the 28S and the 5S rDNA genes (IGS 1) were compared for three European isolates belonging to three Puccinia striiformis f. sp. tritici races, of different geographic origins and virulence spectra. Two PCR products (IGS 1-1 and IGS 1-2, of 1.3 and 1.1 kb, respectively) were recovered from all spore multiplications. The analysis of IGS 1-1 or IGS 1-2 showed that a low mutational polymorphism existed between the three isolates (from 0.7 to 1.1%) and that each IGS 1 type was characterised by a succession of repetitions. The comparison of IGS 1-1 and IGS 1-2 to each other showed that the polymorphism between the two types of IGS was due to a length polymorphism consisting of a 216 bp insertion/deletion located in the repeated region. Two hypotheses can be proposed to explain the coexistence of two different IGS 1 types within one isolate: either each nucleus of this dikaryotic fungus contains one type of IGS 1 or there are two discrete ribosomal units resulting from recombination in the same nucleus.

The identity of strains identified as Trichoderma aureoviride/Hypocrea aureoviridis was reconsidered. Trichoderma aureoviride was isolated originally from a specimen identified as H. aureoviridis and thus is H. aureoviridis. The morphological and molecular characters of most strains identified as T. aureoviride differ from those of the ex-type but are more typical of T. harzianum, a member of sect. Pachybasium. Molecular data do not support inclusion of T. aureoviride in sect. Trichoderma, nor was there strong phenotypic similarity between H. aureoviridis and H. rufa. In the ITS phylogeny the T. aureoviride ex-type and other collections of H. aureoviridis form a strongly supported clade that is separate from any other recognized section of Trichoderma. Hypocrea vinosa, which was originally included in the T. aureoviride aggregate species concept, is distinct from T. aureoviride, but closely allied with H. rufa/T. viride. Trichoderma aureoviride/H. aureoviridis is a rare species, restricted to the UK and the Netherlands. We redefine T. aureoviride, limiting it to strains with very slow growth rate, effuse conidiation, and the ITS-1 and 2 sequence type D.

Twenty-seven Fusarium oxysporum isolates were studied by RAPD-PCR and isozyme analyses. Thirteen isolates were from barley and the rest from different hosts, most of which were dicotyledonous plants. All isolates could be distinguished from each other by RAPD-PCR analysis, and clustered into seven groups in NJ and parsimony consensus trees. Isozyme analysis detected polymorphism in five of the six enzymes and the isolates could be divided into 26 different electrophoretic groups. Five groups were supported by high branch support and bootstrap values in the approximate support tree of combined RAPD-PCR and isozyme data. These five groups were found also in NJ and parsimony consensus trees. The matrices from RAPD-PCR and isozyme data proved to be incongruent, but they did not totally disprove each other. Some correlation was found between geographical origin and phylogenetic relationships of isolates collected from barley. Representatives of the main clades of phylogenetic trees, were further studied by rDNA RFLP and rDNA sequence analyses, together with isolates of other Fusarium species. Isolates of F. oxysporum and F. avenaceum formed distinct groups in the phylogenetic analyses, except for two isolates of F. oxysporum which were grouped with isolates of F. redolens.

Five ophiostomatoid taxa have been found associated with the infructescences of Protea species, an ancient group of flowering plants endemic to South Africa. Two of these fungi are characterized by their unusual Knoxdavesia anamorphs and have been placed in Gondwanamyces. The three remaining species have Sporothrix anamorphs and have accordingly been accommodated in Ophiostoma. The phylogenetic relationships between the fungi associated with Protea spp. and other ophiostomatoid fungi in Ceratocystis and Ophiostoma are unknown. Large subunit ribosomal RNA sequence data was obtained for the fungi associated with Protea infructescences as well as for the type species of Ceratocystis and Ophiostoma. Both groups of ophiostomatoid fungi were phylogenetically distinct from either Ceratocystis or Ophiostoma, despite sharing morphological and physiological characters with these genera. The species of Ophiostoma associated with Protea infructescences group within the Ophiostomatales while species of Gondwanamyces group within the Microascales. Furthermore, the Ophiostoma spp. from Protea should reside in a separate genus and are a fascinating example of convergent evolution towards insect dispersal.

The name Marchandiobasidium aurantiacum gen. sp. nov. is introduced for the teleomorph of Marchandiomyces aurantiacus. Dolipore septa and septal pore caps of the closely related Marchandiomyces corallinus are typical of the Ceratobasidiales, and the basidiomatal characters of Marchandiobasidium aurantiacum are reminiscent of those of the monotypic genus Waitea. Morphological, ultrastructural and molecular data suggest that Marchandiobasidium should not be included in Waitea, but should be treated as a distinct genus.

Rabbit pellets collected from the field were colonized by Podospora pleiospora at the exclusion of other coprophilous fungi, suggesting antibiosis. In liquid culture, P. pleiospora produced sordarin (1); sordarin B (2), a new compound in which sordarose is replaced by rhamnose; hydroxysordarin (3); and sordaricin (4). The major compounds 1 and 2 exhibited minimum inhibitory concentrations of 0.5–2.5 μg ml−1 against the yeasts Nematospora coryli and Sporobolomyces roseus, but showed little or no activity against bacteria or coprophilous filamentous fungi. In liquid culture, the production of 1 and 2 together amounted to 2.7 μg ml−1, whereas in rabbit dung only 1 was produced at a similar concentration (2.3 μg g−1 fresh weight). The biosynthesis of these substances was unaffected by the presence of inoculum of other fungi tested (Sporobolomyces roseus or Penicillium claviforme) in liquid culture or on dung. Sordarin-type natural products are therefore synthesized by P. pleiospora at sufficiently high concentrations to account for antibiosis against yeasts, but not against filamentous fungi.

During the course of a survey of sapstaining fungi in Korea, a Leptographium was isolated from Pinus densiflora and P. koraiensis logs, infested with the bark beetle Tomicus piniperda. The fungus grew optimally at 25 °C on 2% malt extract agar and showed a high level of tolerance to cycloheximide. The Leptographium has unusually short conidiophores and is morphologically similar to L. pini-densiflorae, L. lundbergii, L. yunnanense, and the Leptographium anamorph of Ophiostoma crassivaginatum. Comparisons of DNA sequence data for three gene regions, as well as morphological characteristics, confirmed that this fungus represents an undescribed taxon. We consequently provide the name Leptographium koreanum sp. nov. for it here.

Pythium oligandrum is a parasite of cultivated Agaricus bisporus. Infection results in significant yield reductions and a disease referred to as ‘black compost’. In this study, P. oligandrum isolates were isolated from New Zealand mushroom composts, and their ribosomal DNA internal transcribed spacer (ITS) regions were amplified using the polymerase chain reaction (PCR). ITS nucleotide sequences obtained from New Zealand P. oligandrum isolates were compared with those previously identified P. oligandrum isolates and 23 described Pythium species. Although New Zealand P. oligandrum isolates had high ITS nucleotide identity with internationally identified P. oligandrum, the order of nucleotides in some regions varied when compared with other Pythium species. These varied nucleotides within the ITS region were used to design PCR primers (P.OLIG.F1 and P.OLIG.R04) for the specific amplification of a 384-bp fragment from P. oligandrum DNA. P.OLIG.F1 and P.OLIG.R04 were used to identify a major source of P. oligandrum inoculation on a New Zealand mushroom farm. Application of this diagnostic test will assist farming strategies implemented to prevent future P. oligandrum outbreaks. Furthermore, results presented identify a need for species resolution between P. oligandrum and P. hydnosporum.

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.

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.

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.

This study aims to clarify taxonomic relationships within the current concept of Xanthoria parietina in northern Europe. For comparison, X. calcicola was also included in the study. Morphological as well as molecular data were utilized. Morphology indicated the presence of three species, Xanthoria parietina, X. calcicola, and X. aureola, the latter of which is resurrected here from synonymy. The most important separating characters involve colour and thickness of the thallus, lobe width, morphology of laminar structures, and the texture of the upper surface. X. aureola, as recognized here, mostly occurs on seashore rocks. Part of the IGS region as well as the complete ITS were sequenced in 70 individual thalli representing ten geographical regions in Europe. In total, 19 different IGS haplotypes and 20 different ITS haplotypes were present in the data set. Owing to indications of possible recombination between the IGS and the ITS, the two data sets were analyzed separately. Haplotype networks were estimated, both of which indicate that X. parietina is distinct from X. aureola and X. calcicola. In our sample, the two latter do not share haplotypes, but are only separated by a few mutational steps.

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.