Two distinct and unrelated groups of ophiostomatoid fungi are associated with Protea infructescences. Two species, Gondwanamyces proteae and G. capensis, have unique anamorphs that reside in Knoxdaviesia. This anamorph genus is infrequently found amongst species of Ceratocystis sensu lato. The second group includes Ophiostoma splendens and O. protearum that have Sporothrix anamorphs typical of Ophiostoma. During recent collections of Protea gaguedi infructescences from the Northern Province in South Africa, an apparently new species of Ophiostoma with a Sporothrix anamorph was discovered. It can be distinguished from O. splendens and O. protearum based on its short ostiolar hyphae and its associated plant host. This teleomorph is described as O. africanum and the anamorph as S. africanum. Five species of ophiostomatoid fungi are now known from Protea infructescences and a key is presented for them.

Cercospora leaf spot of olives is a serious defoliating disease attributed to Pseudocercospora cladosporioides. Although the disease is well distributed throughout olive growing regions of the world, its epidemiology and population structure remains unknown. The aim of this study was to establish the genetic variability of Spanish isolates of P. cladosporioides using DNA sequence data from the ITS region, as well as two protein-coding genes, actin and calmodulin. Phylogenetic data obtained here support P. cladosporioides as closely related to other Pseudocercospora species that cluster within Mycosphaerella. Spanish isolates clustered in two clades: isolates from Catalonia were different from those collected in Andalusia. However, isolates appeared to be genetically relatively homogeneous, suggesting that chemical control of this disease via a managed spraying programme may prove a viable option for controlling the disease in Spain.

Clonal isolates of Uncinula necator, the grapevine powdery mildew pathogen, were characterized by mating type, RFLP and PCR analyses. Five random genomic clones, consisting of three multi-copy and two low-copy sequences, were used as probes for hybridization. The oligonucleotides (CAC)5, (GACA)4, the core sequence of the M-13 minisatellite region and the consensus region from a plant intron splice junction (R1), were used as primers in PCR reactions. The DNA probes detected high levels of genetic diversity in U. necator, identifying 34 unique haplotypes among the 81 isolates analysed, with a Shannon diversity index of D′=0·85. In comparison, 15 haplotypes were resolved in PCR reactions with the four primers, resulting in a diversity index of D′=0·59. The overall diversity within the collection, by combining RFLP and PCR data, was D′=0·88, distinguishing 37 haplotypes. The RFLP analysis provided greater discrimination of individual haplotypes than did PCR. Phenetic analysis of the RFLP and PCR data separately and in combination revealed two broad phenetic groups. The average overall similarity between the two groups was 40%. Mating of isolates from the two phenetic groups resulted in viable ascospores. The combination of RFLP, PCR and mating techniques was effective in determining levels of diversity and provided insight into the genetic relatedness of U. necator isolates.

The ability of Peridermium pini and Cronartium flaccidum aeciospores and mycelium to infect known (Vincetoxicum spp., Pedicularis spp., Paeonia spp.) and potential (Melampyrum spp., Pyrola sp., Dactylorhiza sp., Solidago sp., Salix sp., Geranium sp. and Maianthemum sp.) alternate hosts was tested. None of the mycelial cultures and only 10% of the aeciospore samples produced uredinia or telia on the tested species suggesting that most aeciospores in Finland belong to the autoecious P. pini. Aeciospores from three locations in northern Finland, however, produced uredinia or telia on Vincetoxicum mongolicum, V. nigrum, V. fuscatum, Paeonia anomala, three P. officinalis cultivars, Melampyrum sylvaticum and Pedicularis palustris either in vitro or in vivo, indicating that these aeciospores belong to the heteroecious C. flaccidum, which occurs sporadically in Finland. Interestingly, the host-specificity of C. flaccidum encountered in Finland was very low (e.g. one sample produced uredinia or telia on eight species). This, added to the wide distribution of Melampyrum spp. over northern Fennoscandia, suggests that the main alternate hosts for C. flaccidum in Finland may be in Melampyrum rather than Pedicularis.

Comparison of type specimens of A. gerdemannii and A. appendicula revealed the spores to be identical in organization and subcellular structure, indicating they are conspecific. The Glomus morph of A. appendicula was not assigned a name in the protologue, but 11 living cultures started from or producing spores of A. gerdemannii also generated Glomus-like spores of identical morphology to those in type specimens of Glomus leptotichum. These spores were indistinguishable from those in type specimens of G. fecundisporum, indicating they are conspecific. Both species were described in the same paper, so the name G. leptotichum is given priority because type specimens are in better condition and specimens from a reference culture have been identified by an author of both prologues. Absence of a teleomorph and confirmation of dimorphism from single and multiple-spore inoculations of host plants in culture provide the basis for designating A. gerdemannii and G. leptotichum as synanamorphs of one vesicular-arbuscular mycorrhizal fungus.

Erynia neoaphidis is the most widespread aphid pathogen in temperate regions. A few authors have identified this species as infecting some non-aphid hosts. It exhibits high variability with respect to morphology and cultural characteristics that underlines the need for assessment of the genetic variability within this species. We used size of PCR-amplified ribosomal DNA internal transcribed spacer (ITS) coupled with RAPD-PCR pattern in order to investigate genetic variability within a sample of 30 isolates identified as E. neoaphidis and one isolate of E. kondoiensis from diverse countries and hosts. Isolates fell into four discrete ITS size groups: 575 base pairs (bp) in isolates from non-aphid hosts, 1000 (including the E. kondoiensis isolate), 1100, or 1450 bp in those from Aphididae. The 1100-type seems to be the one of most widespread, since two-thirds of the isolates from diverse locations belong to this type. ITS types can be correlated with morphological and physiological characteristics. A great deal of genetic variability exists within ITS types, as shown by RAPD-PCR. RAPD groupings could be related with geographical origin in some cases. There was no apparent relationship between host and ITS type or RAPD pattern among isolates from Aphididae. PCR-based technologies provide evidence that could be used to clarify the taxonomy of E. neoaphidis and closely related species, and will help identify markers related to epidemiological characteristics.

The phylogenetic position of Uncinula forestalis within the Erysiphales has been inferred from 5.8S, 18S, and 28S rDNA sequences. Although the appendages of the ascomata are Uncinula-like, i.e. unbranched with curved-coiled apices, U. forestalis is situated at the very base of the large Erysiphales cluster, far from the ‘Pseudoidium’ clade (Erysiphe, including Microsphaera and Uncinula) and separate from the recently introduced basal genus Parauncinula. U. forestalis differs morphologically from the species of Erysiphe sect. Uncinula in having terminal, fasciculate (as in Podosphaera tridactyla), septate ascoma appendages and a Euoidium-like anamorph (conidia catenate). In Parauncinula, the appendages are also terminal but not fasciculate, the ascospores are curved, and the anamorph is lacking. Uncinula forestalis is a basal, tree-inhabiting powdery mildew with some additional ancestral characteristics, including Uncinula-like appendages and 6–8-spored asci. The new genus Caespitotheca gen. nov. is described with C. forestalis comb. nov. (syn. Uncinula forestalis) as the type species. We calculated the timing of the divergence of U. forestalis and P. septata using a molecular clock of the Erysiphales (6.5×10−10 changes per site per year in domains D1 and D2 of 28S rDNA) and a 28S rDNA data set. The results suggest that the divergence of U. forestalis and P. septata from other Erysiphales occurred between 90 and 80 Myr ago; i.e. the divergence of the two ancestral species may have occurred in the late Cretaceous.

The ITS region of the rRNA genes was amplified using DNA extracted from Terfezia pfeilii fruit-bodies. Two types of restriction fragment profiles were revealed among 11 fruit-bodies by the Hinf I restriction enzyme. An additional four truffles collected from the same field one year later were found to have a mixed profile combining both types. Germinated spores from one of these mixed-profile individuals had a single profile belonging to one or the other of the two types. Roots collected around and underneath one Terfezia fruit-body were traced to different plants. Roots of a watermelon plant (Citrullus vulgaris) showed a non-vesicular-arbuscular endomycorrhizal association. Restricted ITS amplification fragments of mycorrhized roots from C. vulgaris were shown to match T. pfeilii fruit-body ITS-RFLP fragments. The significance of this finding with regard to T. pfeilii potential host range is discussed.

The cultivated mushroom Agaricus bisporus was grown and fruited in wheat straw compost. RNA was extracted from samples of mycelium growing in compost during colonization and fruit body development. Laccase gene (combined lcc1 and lcc2) expression and cel3 gene (cellobiohydrolase) expression were determined by Northern blot analysis and by competitive RT-PCR. The level of laccase transcripts was highest in colonized compost (prior to the onset of fruiting), declined during the period of fruit body enlargement and increased again after harvesting and during the second flush of fruit body production. No transcripts of the cel3 gene were detectable in colonized or pre-fruiting cultures, but the level became detectable and rose to a maximum at the veil-break stage of fruiting, declined after harvesting and rose again during the second flush. Gene expression for laccase and cellobiohydrolase correlated with the known changes of laccase and cellulase enzyme activities during the fruiting life cycle.

We describe a 16 yr dataset of ectomycorrhizal fruit bodies under Scots pine (Pinus sylvestris), starting from seedlings, and explore the effects of a 50% thinning treatment imposed in year 12. The over-riding pattern in the data was of successional development, with Paxillus involutus and Laccaria proxima in the earliest years, followed by Suillus species, while in later years Amanita and Cortinarius species became prominent. The typical pattern was for each species in turn to increase to a maximum count then gradually decline without actually disappearing from the community (an addition succession). For nine out of the 30 species recorded the time profiles were fitted significantly by a log-normal curve, although the best-fit lognormal models consistently under-estimated the peak count. Thinning increased counts of Suillus bovinus, Gomphidius roseus and Cortinarius semisanguineus, but overall its effect on the community was minor. Rainfall in September was weakly correlated with successional advancement (measured as DCA first axis scores). Mean fruiting date increased significantly as the stands aged.

Pseudotulostoma volvata gen. sp. nov. is described from the south-central Pakaraima Mountains of Guyana. Pseudotulostoma volvata is associated with ectomycorrhizal Dicymbe corymbosa trees (Caesalpiniaceae) and placed in the Ascomycota, Eurotiales, Elaphomycetaceae. Included are a description of the genus and species, illustrations of the macroscopic and microscopic features, and a discussion of the distinctive features and phylogenetic placement of this fungus.

Ribosomal DNA sequences were assessed for their usefulness in distinguishing among Trichoderma isolates and for their robustness in resolving their phylogenetic relationships. DNA sequences from the D2 region of the 28S rRNA gene were determined for 50 Trichoderma isolates representing seven species. Eight distinct sequence types existed, and were mostly consistent with groupings based on morphology. Sequence variability within the D2 region alone was not sufficient to provide a reliable phylogeny. Sequences from the ITS1, 5.8S and ITS2 regions were subsequently determined for 18 of the isolates. Eight distinct ITS sequence types were detected among these 18 isolates. The ITS sequence types were generally consistent with morphology, ITS1 sequence data supported the identification of the Th3 T. harzianum group of Muthumeenakshi et al. (1994) as T. atroviride. The data also confirmed that the biocontrol strains of this study were different from those causing disease problems in the mushroom industry in Europe and North America. Results from the phylogenetic analysis stress the importance of testing the robustness of data used to predict phylogenies. Two ITS sequence data sets for the same group of isolates produced significantly different phylogenies. Congruence analysis detected that T. inhamatum (GJS90-90) was corrupting tree topologies and ‘first order pruning’ was performed by removing its sequence from the two ITS data sets. Subsequent differences in the topologies of pruned ITS1 and ITS2 trees were attributed to a lack of phylogenetic information in the ITS2 sequence region. Although ITS sequences successfully differentiated among morphologically distinct isolates within Trichoderma, it did not provide a sufficient phylogenetic signal to resolve all of their relationships.

Cryphonectria canker caused by Cryphonectria cubensis is one of the most destructive diseases of Eucalyptus plantations in South Africa. To implement a meaningful management of plantation diseases, it is important to have an understanding of the population diversity of the pathogen. In this study, trees were surveyed to determine whether C. cubensis reproduces sexually in South Africa. The diversity of the South African C. cubensis population was assessed based on vegetative compatibility tests. Field inoculations were used to determine whether VC groups correlated with virulence. Only pycnidia were found on cankered trees, indicating that sexual reproduction does not occur. Only 23 VC groups were found amongst 100 isolates each collected from single diseased trees. A low degree of genetic diversity also indicated that sexual reproduction is absent or rare in the South African C. cubensis population. Inoculation studies revealed that isolates belonging to different VC groups differ significantly in their ability to cause lesions. The low level of genetic diversity enhances opportunities to capitalise on hypovirulence to reduce the impact of the pathogen in the future. It also supports the view that the fungus was recently introduced into South Africa.

This study aimed at testing the hypothesis that the genetically distinct Tuber aestivum population on the island of Gotland, Sweden, is adapted to habitats different from French T. aestivum populations. The soil structure, soil chemistry, bedrock, climate, vegetation and host tree continuity of 18 T. aestivum sites on Gotland were analysed and compared with data from France. We conclude that T. aestivum can grow in soils with a broad soil structure range and that no striking differences in soil chemistry were found. No T. aestivum indicator plants other than the host trees were found, but the host tree continuity on the T. aestivum sites on Gotland was more than 300 yr. If the T. aestivum population on Gotland constitutes an ecotype it is rather an adaptation to the colder and drier climate on Gotland. Selecting local T. aestivum inoculum for truffle orchards in northern Europe could be important for successfull truffle production.

In the mushrooms Pleurotus ostreatus and Agrocybe aegerita, expression of the hemolytic proteins ostreolysin and aegerolysin, which belong to the aegerolysin family, has been shown to be initiated specifically during formation of primordia and fruiting bodies. We used rabbit anti-ostreolysin and fluorescent rhodamine-labelled secondary goat antibodies for monitoring ostreolysin and aegerolysin in situ during the mushrooms' development. In parallel, the protein level in developing tissues was monitored with SDS-PAGE and hemolytic assay. Immunolocalization of ostreolysin, visualized by epifluorescence, together with biochemical tests, confirmed specific expression of ostreolysin and aegerolysin in the primordia and fruiting bodies but not in the vegetative mycelia. In the primordia, the proteins were disposed diffusely. In growing and mature fruit bodies they persisted in the lower part of the pileus, in particular in basidia and basidiospores, while in other parts only a few focal remains were observed. Confocal microscopy of immunolabelled sections showed that intracellular ostreolysin was located specifically along the inner edges of hyphae. Since both proteins were found preferentially in the rapidly growing primordia, and in the basidia and basidiospores of maturing fruit bodies, it is suggested that they might play a role in the processes of fructification and/or sporulation.

Pulse-field gel electrophoresis (PFGE) was used to determine the genome size and characterize karyotypic differences in isolates of the cacao biotype of Crinipellis perniciosa (C-biotype). The karyotype analysis of four isolates from Brazil revealed that this biotype could be divided into two genotypes: one presenting six chromosomal bands and the other presenting eight. The size of the chromosomes ranged from 2.7 to 5.3 Mb. The different genotypes correlate with telomere-based PCR analysis. The isolates with six chromosomal bands had two that appeared to be doublets, as shown by densitometric analysis, indicating that the haploid chromosome number for this biotype is eight. The size of the haploid genomes was estimated at approximately 30 Mb by both PFGE and Feulgen-image analysis. DNA hybridization revealed that the rDNA sequences are clustered on a single chromosome and these sequences were located on different chromosomes in an isolate dependent manner. This is the first report of genome size and chromosomal polymorphism for the C-biotype of C. perniciosa.

This study aimed to test whether fungal community on moose (Alces alces) dung is affected by habitat. We used dung of homogenous origin, composition, and age. Dung was placed in three different habitats in north-eastern Sweden, and was checked again after 35–36 d. Of the 26 species of fungi found, 12 were new to the region, 17 had never been observed on moose dung, and two were not previously described. We found a significant difference in species composition between the habitats, with a low number of species in the spruce forest and about a threefold increase in the pine forest and the open mire. Species diversity was negatively associated with degree of insect attack. This suggests that insects feeding either on the dung or the fungi (spores, mycelia) may be an important factor explaining the observed pattern. In order to test this hypothesis we need to run experiments excluding insects.

Mycobiont isolation experiments were performed on six species of Ramalina from Brazil: R. celastri, R. complanata, R. dendriscoides, R. gracilis, R. peruviana and R. sprengelii. This study aimed to optimize the culture conditions and nutrient requirements of the selected mycobionts. The aposymbiotic R. complanta was successfully isolated from ascospores, while aposymbiotic R. peruviana was obtained from thallus fragments. In R. peruviana the production of secondary metabolites was investigated under aposymbiotical growth conditions using HPLC. When cultivated on solid medium, this mycobiont produced the typical chemosyndrome (sekikaic acid and satellite compounds), found in the voucher lichen thallus. When cultivated in liquid medium (immersed in malt yeast medium in the absence of agar), only one, the major lichen substance, sekikaic acid, was synthesized by the fungus. In addition, atranorin was formed, but was not detected in any of the voucher specimens. Red pigments were found in solid and liquid cultures. These were separated into two compounds, but could not be fully identified. R. celastri spores germinated, but did not form mycelia. R. dendriscoides, R. gracilis and R. sprengelii were not successfully cultivated in aposymbiotic conditions, although eight different culture media were tested.

Osmotrophic eukaryotes with a cell wall during the assimilative phase are ‘fungi’: the term ‘fungus’ is an essentially physiological concept, thus flagellate fungi conforming to this definition are not ‘pseudofungi’. But mycologists may also work with flagellate organisms which are phagotrophic or lack a plasmodial cell wall and, therefore, are not strictly ‘fungi’. In fungi the flagellate stage is confined to planonts (asexual zoospores and gametes).

Flagellar form and function have many conserved characteristics, but although ultrastructural diversity, particularly in kinetosome/flagellar root structure, has been intensively studied, other morphological features of the zoospore have been less thoroughly explored. The numbers, lengths, orientations and structural ornaments of flagella all provide data for biodiversity assessments at the species and ecological levels. The structural and functional biodiversity of fungal flagella and zoospores are reviewed, particularly with respect to the isokont/anisokont flagellar lengths and the isokont/heterokont flagellar ornaments of the zoospore. The straminipilous ornamentation has particular functional significance.

Correlations between molecular biology and flagellar ultrastructure indicate that several independent phylogenetic lines have evolved flagellate fungi. The strengths and weaknesses of the databases for these phylogenies are discussed in historical and current contexts.