Genetic variation within a collection of 25 isolates of Monascus from red rice and sofu was assessed with RAPD markers using genetic distances (1-Jaccard's coefficient) calculated for all combinations of isolates. Cluster analysis based on genetic distance was performed using the Ward's minimum variance method. Five distinct clusters were revealed based on genetic distances between and among clusters. The robustness (reproducibility) of the cluster assignments was tested by resampling (bootstrap) analysis. Cluster distribution was visualized as a three-dimensional graph based on multiple correspondence analysis. A dendrogram, based on the clusters, was constructed to examine the relationships. Three clusters accounted for 21 isolates; the fourth cluster consisted of three isolates which were quite distinct from each other and all other isolates. The fifth cluster, a single isolate from Japan, was very different from all others in RAPD patterns and was used as an outgroup. Resampling analysis indicated that the 25 isolates represented four genetic lineages of red rice fungi, suggesting that a relatively narrow genetic source of Monascus isolates is used in food products in Asia.

Compared with other Armillaria species, natural melanized rhizomorphs of A. tabescens are rarely observed. Growing in autoclaved Vitis stem segments, A. tabescens isolates from the Ozark Mountains in the central USA formed fully melanized rhizomorphs, thinner and shorter than those observed for other Armillaria species under field conditions, and only under conditions of both high oxygen availability, [ges ]2 μg cm−2 min−1 and moisture near saturation. Conducive conditions were used to compare the rhizomorph generation capacities of A. tabescens, A. gallica, and A. mellea, which have overlapping host and geographic ranges in central North America. While the rhizomorphs of A. tabescens were significantly shorter than those of the other two species, A. gallica and A. tabescens produced similar numbers of rhizomorph initials. Finally, we demonstrated the ability of melanized A. tabescens rhizomorphs to span woody food bases and thereby establish viable infections by penetration of intact bark of Vitis stem segments. We hypothesize that A. tabescens rhizomorphs form under conditions of periodic saturation which promote rapid water movement through naturally occurring lacunae in the soil. Thus, the role of A. tabescens rhizomorphs in the spread of the fungus should be re-evaluated.

RAPD amplifications of 119 Mexican Colletotrichum gloeosporioides isolates from wild native Stylosanthes plants, of two Australian (internal reference isolates of type A and type B) and of two African (previously demonstrated as different from the A and B types) allowed the generation of four polymorphic clusters. The RAPD cluster I contained the Australian type A and 103 Mexican isolates from other species than S. guianensis. The cluster II contained the Australian type B isolate and 14 Mexican isolates from S. guianensis. The clusters III and IV contained one African and one Mexican isolate each.

A phylogenetic study was performed using the ITS1 sequences from 32 representative Mexican isolates selected in the four RAPD clusters, from the two Australian and the two African used as references, from 13 C. gloeosporioides from various hosts and origins available in the EMBL databank and from three phylogenetically related species of Colletotrichum. Two clusters (S and VHO) were generated. Cluster S contained the 30 isolates clustered in RAPD I and II, and the type A and B isolates from Stylosanthes selected in EMBL. Cluster VHO contained the two isolates belonging to RAPD clusters III and IV as well as the remaining Colletotrichum spp. and C. gloeosporioides isolates from various hosts and origins selected in EMBL.

Severe type A lesions were observed on several Stylosanthes species, including S. guianensis, after inoculation with the 26 Mexican isolates selected in RAPD cluster I. The four Mexican isolates selected in RAPD cluster II induced extensive type B lesions only on S. guianensis genotypes. The virulence spectra of the isolates was not linked to variations in their ITS1 region sequence. A wide range of anthracnose lesions was induced by the Mexican isolates in interactions with the various Stylosanthes genotypes.

On the basis of results obtained from the molecular and pathogenic experiments in the present study, of morphological and molecular data from previous work, as well as from other authors, a taxonomic infraspecific differentiation was proposed, as C. gloeosporioides f. stylosanthis f. sp. stylosanthis and f. sp. guianensis.

Conidial germination in Neurospora crassa was inhibited by the anticalmodulin (CaM) agents calmidazolium (CaMZ) and trifluoperazine (TFP), indicating the importance of CaM-dependent events during vegetative growth. An increase in intracellular CaM concentration, observed during conidial germination, was associated with enhanced incorporation of [3H]phenylalanine into CaM and increased CaM-dependent protein kinase activity. Phosphorylation of two proteins (71 and 25 kDa) during conidial germination, and of four proteins (71, 47, 38 and 32 kDa) during hyphal elongation was noted to be CaM-dependent since these phosphorylations were inhibited in vitro by CaMZ. The possible regulatory functions of CaM during conidial germination are discussed in relation to phosphorylation of specific proteins during vegetative growth in N. crassa.

The phylogeny of the Dermatocarpon miniatum-complex (Verrucariales, lichenized Ascomycota) was studied using nuclear ITS sequence data by both parsimony and Bayesian inference of phylogeny. The ITS region contains a substantial amount of variation which resolves the relationships of terminal groups, while the more basal clades have low support in the analyses. D. miniatum var. miniatum and var. complicatum are polyphyletic, while var. cirsodes is monophyletic but located within the complex, as are both D. leptophyllum and D. linkolae. The variation within the D. miniatum-complex is significantly greater than that between some transatlantic species such as D. luridum and D. meiophyllizum. The new names D. taminium sp. nov. from the Greater Sonoran area, and D. tenue comb. nov. (syn. D. muehlenbergii var. tenue) are introduced.

Byssosphaeria, Chaetosphaeria and Niesslia are discussed and illustrated in relation to their occurrence on palms. New collections of B. schiedermayeriana are reported including Chaetosphaeria exima, which is a new synonym. Chaetosphaeria arecacensis, C. hongkongensis and C. palmicola spp. nov. are described and new collections of C. callimorpha are reported. Cryptophiale is reported as a new anamorph connection for Chaetosphaeria. Niesslia palmicola sp. nov. is described and new collections of N. exosporioides are recorded. Ornatispora gen. nov. is introduced to accommodate several Chaetosphaeria-like species from palms. In Ornatispora, ascomata are superficial, black, papillate, smooth-walled or covered in setae and paraphyses are filiform, non-septate and deliquesce in dried material. Asci are 8-spored, clavate, and lack an apical apparatus and deliquesce at maturity, while ascospores are ellipsoidal, 1-septate, hyaline, verrucose and surrounded by a mucilaginous sheath. One species is closely associated with Didymostilbe aurantiospora, which is considered to be the anamorph. Three new species of Ornatispora, and one new combination are described and illustrated. Ornatispora is compared with Bertia, Chaetosphaeria, Peethambara and Niesslia.

The taxonomy of the marine species of Collemopsidium in northwest Europe was investigated using morphological and molecular evidence. 210 specimens were collected from the west coasts of Norway and Ireland, and morphological and ecological variables recorded. ITS1 rDNA sequences were obtained from 24 specimens. A phylogenetic analysis, resulting in a single optimal tree, was performed under the unweighted least squares optimality criterion based on maximum likelihood distances obtained from unaligned sequences. Principal components analysis (PCA) was performed on morphological variables of the sequenced specimens, and classification was carried out by maximizing agreement between the phylogenetic tree and the PCA. Thallus immersion, and perithecial immersion and size, were the most important characters for discriminating between taxa. Apart from substratum, niche separation between taxa was small but statistically significant as shown by a redundancy analysis (RDA). Variance partitioning indicated that genetic variation is vastly more important than ecology for explaining phenotypic variation. Five species of marine Collemopsidium are recognized, including two new combinations: C. foveolatum (syn. Arthopyrenia foveolata) and C. ostrearum (syn. Lecanactis ostrearum) combs. nov.

The ear rot severity of 15 maize hybrids and the accumulation of fumonisin B1 (FB1), fumonisin B2 (FB2), beauvericin (BEA) and fusaproliferin (FP) after artificial inoculation in the field with a toxigenic strain of Fusarium proliferatum has been investigated in Poland during the seasons 1996, 1997 and 1999. The year of inoculation proved a significant influence on ear infection degree and mycotoxin accumulation. Inoculated ears contained 11–71% Fusarium-damaged kernels. Mycotoxins were detected in all hybrids at levels of 18–231.9 μg g−1 for FB1, 0.4–26.5 μg g−1 for FB2, 0.2–19.6 μg g−1 for BEA and 0.3–6.4 μg g−1 for FP. Mycotoxin concentrations were higher in Fusarium-damaged kernels (up to 361.5, 41.1, 44.3 and 10.0 μg g−1 for FB1, FB2, BEA and FP, respectively) than in healthy-looking kernels (up to 26.0, 2.3, 1.9, 0.3 μg g−1 for FB1, FB2, BEA and FP, respectively). All hybrids showed high susceptibility to the fungal infection and high toxin content in kernels during the three years of investigation.

The forest pathogen Dothistroma pini, which produces the polyketide toxin, dothistromin, was transformed with the pAN7-1 plasmid containing the Escherichia coli hygromycin B phosphotransferase gene (hph). The frequency of hygromycin resistant transformants was 9–48 μg−1 DNA. Southern blot analysis indicated that a single copy of the vector DNA had integrated into the genome in at least six of the eight transformants tested and that the site of integration was different in each case. All transformants analysed were mitotically stable through several subcultures without selective pressure.

The external morphological changes occurring during initiation and early stages of fruit body development of a Morchella sp., before the development of asci, were examined by scanning electron microscopy. Four stages of primordial development were distinguished. First, disk-shaped knots of 0.5–1.5 mm were observed on the surface of the substrate. Next, the knot inflated and a primordial stem emerged from its centre. Afterward, the stem lengthened, oriented upward, and two types of hyphal elements developed: long, straight and smooth basal hairy hyphae and short stem hyphae, some of which were inflated and projected out of a cohesive layer of tightly packed hyphal elements. Finally, when the stem was 2–3 mm long, pre-apothecia emerged in the apical end, with ridges and pits having distinguished types of paraphyses. Extracelluar mucilage covered the ridge layer and helped give the tissue its shape and rigidity.

Development of specific DNA probes targeting rDNA internal transcribed spacer (ITS-1 or -2) sequences is described for the detection of strains representing uninucleate and binucleate Rhizoctonia spp. and Suillus bovinus. Discriminatory taxon/species-specific target sequences were identified following full length ITS sequence alignment of the test fungal sequences and those of other root associated pathogenic or mycorrhizal fungi. Both long (124–151 bp) and shorter (20–25 bp) probes were generated for assessment in Southern dot blot and liquid hybridization ITS capture-fragment length polymorphism assays. Fungal genomic DNA was presented as the target in dot blot protocols using the longer DIG (digoxigenin) labelled probes whilst the shorter 3′ biotin-labelled oligonucleotide probes were hybridized to PCR amplified full length ITS (ITS1-5.8S-ITS2) in both dot blot and liquid hybridization assays. The optimal hybridization temperatures for dot blot detection also produced maximal target specific signals in the liquid hybridization protocol. The latter protocol was found to be more discriminatory as target fungi were detected on the basis of combined probe hybridization-ITS capture and 5′ Cy-5 labelled ITS length polymorphism analysis (±5 bp) following denaturing sequencing gel electrophoresis in a ALFexpress DNA sequencer.

Species belonging to Hypoxylon and related genera are bark pathogens and wood decay agents of angiosperms, sometimes able to live asymptomatically in host tissues. In the present work the ITS regions of the ribosomal DNA from 21 isolates belonging to five species were sequenced and then compared with each other. A phylogenetic parsimony analysis among isolates was carried out. ITS2 appears to be the most informative among the sequenced portions of rDNA. Results are strictly consistent with the latest taxonomic arrangements obtained with classical morphological approaches. The ITS sequence analysis is an efficient and accurate method for discriminating among taxa of Hypoxylon and related genera.

Pseudohelicomyces albus gen. et sp. nov. is described. It is characterised by the presence of hyaline, cylindrical arthroconidia originating from either spiral vegetative hyphae or from spiral, branched conidiophores. The hyphae and conidiophores may or may not possess clamp connections. The characteristics of the basidiocarps formed in culture were identical to material collected from nature. P. albus is the anamorphic stage of Psilocybe merdaria and shows similarities to Helicomyces.

The correct identity of the ergot strains Claviceps fusiformis PRL 1980 (ATCC 26245), C. fusiformis 129 and 59, and Pepty 695/ch-I, formerly described as Claviceps purpurea, was proven by means of RAPD, genomic DNA restriction patterns as well as by a literature search about the strain origin.

Candelabrum desmidiaceum sp. nov. and C. clathrosphaeroides sp. nov. are illustrated and described. Morphologically, they represent a new type of conidium development previously unknown in Candelabrum: the formation of a basal plate by dicho- or trichotomous branching of the conidial body in combination with branches growing upwards at the outer margin of the basal plate which are distinctly verrucose outside, forming a hollow sphere enclosing an air bubble in the centre. In Austria, both species were found mainly on submerged beech leaves, but in the U.S.A., C. clathrosphaeroides was isolated from submerged leaves of many different deciduous species, indicating a wider host range. Both species are aero-aquatic fungi, and C. clathrosphaeroides, at least, is widely distributed in temperate climates. A key to the species of Candelabrum is provided.

Lewia avenicola sp. nov. (anamorph Alternaria sp.) is described. The fungus was isolated from oat grain in Norway. The fungus produced conidia within a month, and ascomata with fully mature ascospores within 8 months following inoculation, when stored on slants of synthetic nutrient agar (SNA) at 4 °C in darkness. A comparison with other known Lewia species is made, and a key to the five known Lewia species is included.

The entomopathogenic Beauveria bassiana is a potential biopesticide. Fungicide-tolerant isolates of this fungus would have an added advantage of being compatible with the conventional chemical methods of pest control. Therefore, 30 isolates of the fungus were screened for tolerance to bavistin a commonly used benzimidazole fungicide containing methyl benzimidazol-2-yl carbamate (MBC). Germination and growth bioassays in the presence of 0.05% bavistin were conducted for screening. Three tolerant isolates were identified, showing tolerance up to 2% bavistin. Mutation in the β-tubulin gene is known to confer tolerance to MBC; the nine known mutation sites in the gene involved were sequenced in the tolerant strains. The β-tubulin gene from codons 1–405 was amplified using two pairs of degenerate primers, designed for the conserved region of the β-tubulin gene and sequenced. From the sequence suitable primers were designed for the regions flanking the nine known sites involved in mutations conferring MBC tolerance. The amplified products with these primers from the MBC-tolerant isolates were sequenced and in two (ARSEF 1315 and ARSEF 1316) a mutation was detected in the 198 codon resulting in replacement of glutamic acid with lysine. In the third tolerant isolate (ITCC 913) no mutation could be detected in any of the nine known sites conferring tolerance to MBC. To complete the sequencing of the β-tubulin gene, the remaining part (from codon 405 onwards) of the gene was isolated by a three-prime gene walk. The gene sequence showed a close homology to fungal β-tubulin genes with four introns.

Colletotrichum dematium is the cause of a new fungal stem disease on cowpea in South Africa. The pre-penetration and infection process of this fungus on cowpea stems was studied by light microscopy and SEM. Conidia began germinating at 6 h post inoculation (hpi), forming appressoria directly or at the ends of germ-tubes. By 14 hpi appressoria had melanized and direct penetration of host tissue had begun. At 20 hpi infection vesicles formed in epidermal cells. Thick, knotted primary hyphae formed from these vesicles and entered adjacent cells. At ±40 hpi C. dematium produced secondary hyphae which were highly branched, and grew extensively inter- and intracellularly. After approximately 48 hpi light brown lesions appeared on the stem, associated with the invasion of secondary hyphae into cells and cell necrosis. Acervuli with one or two melanized setae were visible on lesions by 70 hpi.

The utilization profile of 49 carbohydrates, based on API 50 CH biochemical tests, was used for the identification and the discrimination of 75 isolates of Beauveria and Tolypocladium. The data were used to construct a most parsimonious tree in which Beauveria and Tolypocladium were separated into two main branches. Although the Beauveria isolates could not be separated at the species level, eight different clusters corresponding to the eight Tolypocladium species studied were revealed. The reproducible carbohydrate utilization profiles of B. bassiana, B. brongniartii, T. cylindrosporum, T. niveum, T. geodes and T. tundrense have permitted the construction of an identification key that could serve as a tool to complement the morphological description of selected Beauveria and Tolypocladium isolates for their correct identification at the species level. The API 50 CH method is time-saving and can complement effectively more classical techniques for the discrimination between Beauveria and Tolypocladium and for the taxonomy of Tolypocladium at the species level.