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.

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.

Two specimens of a new species of Hypocrella with large stromata were collected in Bolivia and Costa Rica. The morphology of the new species, H. macrostroma sp. nov., was compared with that of other species with large stromata, i.e. H. africana, H. gaertneriana, and H. schizostachyi. In addition, phylogenetic analyses of partial sequences from three genes, large subunit nuclear ribosomal DNA (LSU), translation elongation factor 1-α (EF1-α), and RNA polymerase II subunit 1 (RPB1), were conducted to determine the relationships of the new species to other species of Hypocrella/Aschersonia. Phylogenetic analyses show that H. macrostroma belongs to a strongly supported clade that includes H. africana, H. schizostachyi, and Aschersonia insperata, whereas other Hypocrella species belong to two sister clades. Hypocrella macrostroma is described and illustrated, and a lectotype is designated for H. gaertneriana.

Twenty-six fungi associated with leaf spots of palms in north Queensland have been identified, including 12 ascomycetes, one basidiomycete and 13 deuteromycetes. Eight were identified as species new to science. A new species of Pseudospiropes is described and descriptions of two unnamed species of Glomerella are also given.

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.

The electrophoretic karyotype (EK) of 24 strains of Botryotinia fuckeliana was resolved to determine the extent of chromosomal polymorphism among them. Based on EK variation, nine different profiles with a number of bands ranging from 5 to 8 were found. Those profiles were not related to the origins of the strains, hosts, or year of isolation. Inheritance of chromosomal bands was studied by analyzing the EK of the progeny from crosses between sexually compatible B. fuckeliana strains that had different karyotype. The EKs of 44 monoascospore strains showed 12 new EK profiles, suggesting that chromosomal rearrangements generated after meiotic recombination provides EK variability in this fungus. Also, we report here that the degree of EK variability indicates that sexual reproduction under natural conditions might be quite important in the life cycle of B. fuckeliana. Hybridization analysis showed that, in most strains, the rDNA is located in both a chromosomal band of high molecular weight and different size depending on the strain and a band of low molecular weight, revealing evidence for chromosome rearrangements in this plant pathogen.

The nematophagous fungus Pochonia chlamydosporia is a potential biocontrol agent against root knot and cyst nematodes. Genetic transformation of the fungus to introduce visual marker genes, novel traits, or changes in expression levels of endogenous genes, would greatly enhance understanding of its behaviour on nematode-infested roots and of its interactions with other soil and rhizosphere microorganisms. A transformation system for the introduction of novel genes into P. chlamydosporia has been developed. Methods to generate protoplasts, introduce DNA and regenerate transformed viable fungal mycelium have been optimised, using plasmids carrying the green fluorescent protein marker gene gfp and the hygromycin resistance gene hph. Cultures of P. chlamydosporia were resistant to high levels of a range of fungal inhibitors, including hygromycin, that are commonly used with dominant selectable marker genes in the transformation of other fungi. However, regenerating protoplasts transformed with hph could be selected by their ability to grow through an agar overlay containing 1 mg ml−1 hygromycin. Green fluorescence was observed in protoplasts and regenerating mycelium after transformation with gfp, but the GFP phenotype was lost on subculture. Maintenance of introduced genes was not stable, and during subculture, PCR assays indicated that the transformants lost both hph and gfp. When these genes were introduced on the same plasmid, segregation of hph and gfp was observed prior to their loss. It was unclear whether the introduced plasmids were able to replicate autonomously in P. chlamydosporia, or if they integrated transiently into the fungal genome. Possible reasons for the instability of the transformants are discussed.

A thermostable intracellular lipoxygenase from Thermomyces lanuginosus was purified to sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) homogeneity by ion-exchange chromatography on diethylaminoethyl (DEAE)-Sepharose, ion-exchange chromatography on carboxymethyl (CM)-Sepharose and Phenyl-Sepharose hydrophobic interaction chromatography. The molecular weight of the enzyme was estimated to be 100 kDa by SDS-PAGE. The lipoxygenase exhibited maximal activities at pH 6.0. The optimum temperature for the activity was 55 °C. The enzyme was thermostable at 50 ° with half-lives at 60 ° of 20 min and 70 ° of about 7 min. The lipoxygenase activity was completely lost by heating at 80 ° for 5 min. The lipoxygenase catalyzed the oxidation of linoleic acid into 13-L-hydroperoxy-9,11(Z,E)-octadecadienoic acid(13-HPOD) as the major product. The enzyme had an apparent Km of 8.5 μM and Vmax of 10.8 μmol mg−1 min−1 with linoleic acid as substrate.

Unidentified isolates of Cylindrocladium with 1–3-septate conidia and clavate vesicles were compared with C. theae, C. colhounii var. colhounii, C. colhounii var. macroconidiale, C. gracile, C. pteridis and Calonectria gracilis. Ribosomal DNA (rDNA) of these isolates were digested with the restriction enzymes EcoR I, Hind III and Xho I, and Southern analysis performed with the 6·3-kb rDNA repeat unit of Neurospora crassa as DNA probe. Based on differences in general morphology, supported by their rDNA restriction fragment length polymorphisms, isolates of all species and varieties could be distinguished. Furthermore, the Cylindrocladium anamorph of Calonectria gracilis was shown to be distinct from Cylindrocladium pteridis and C. gracile. The name Cylindrocladium pseudogracile sp. nov. is, therefore, proposed for the undescribed anamorph of Calonectria gracilis.

A study on the mangrove leaf litter fungi of two tropical mangroves in Singapore was carried out. Five Halophytophthora species and four isolates which did not fit the descriptions of known species were isolated from leaves of Avicennia alba, A. rumphiana, Rhizophora apiculata and R. mucronata. Halophytophthora exoprolifera, H. kandeliae, H. operculata, H. spinosa var. lobata and H. vesicula are new records for Singapore; the first three also being new records for tropical mangroves. The isolate of H. exoprolifera sporulated at 28 °C and may be a tropical strain. H. operculata is reported for the first time outside of Australia. The results of this study contribute to the little known distribution of the Halophytophthora in the Indian Pacific region. Thirteen deuteromycetes were also isolated from the mangrove leaf litter and represented both phylloplane fungi as well as possible marine fungi.

The relationship between hyphal growth and branching of the grape pathogen Botrytis cinerea was determined on solid media containing either glucose, fructose, sucrose, tartaric acid or malic acid. The concentration of the carbon source had little effect on specific growth rate or the specific rate of tip formation, but growth was inhibited at high concentrations of tartaric and malic acids. Hyphal growth unit length and hyphal extension rate increased with increasing sugar concentration and were always significantly greater than values on tartaric or malic acids. The data provide an explanation for colonization patterns of grape berries. Growth will be poor during the period from setting to the onset of ripening, when organic acids are the main carbon source produced by the berry. Following the onset of ripening, the production of sugars provides more favourable carbon sources for the fungus, enabling achievement of higher specific growth rates, greater hyphal extension rates and, hence, greater colonizing potential.

A new species of Chaetomium, C. cuyabenoensis, is described from the Ecuador rain forest. It is similar to Chaetomium longicolleum but differs by having smaller and, in face view, strongly limonifom to quadrangular ascospores.

A study was designed to isolate and identify the endophytic mycobiota in leaves and twigs of Rubus spectabilis and R. parviflorus, the long-term goal being biological control of these plants with endophytes. Endophytic fungi were isolated from healthy-looking leaves and twigs collected from each species on Vancouver Island, British Columbia, Canada. More than 90% of the leaves of both species were colonised by endophytes. In addition, one third of the twig samples of each species were also colonised. Most isolates belonged to Phomopsis, and five species were identified and characterised according to morphology of the cultures and fructification structures. All Phomopsis species occurred on both Rubus species, but, one primarily colonised R. parviflorus, whereas another preferentially colonised R. spectabilis. The suitability of the isolated endophytes to inhibit growth of R. spectabilis and R. parviflorus, which compete with conifers in reforestation sites, is discussed.

A Cylindrocladium-like hyphomycete collected on leaf litter of Celtis tala in Argentina had rDNA sequence data (ITS and LSU) that showed it resides in the Hypocreales, and is a member of the Nectriaceae, closely related to, but distinct from Cylindrocladium. A new genus, Dematiocladium and species, D. celtidis gen. sp. nov. is, therefore, introduced to accommodate this fungus. Based on morphology, it can be distinguished from other conidial hypocrealean genera with hyaline, penicillate conidiophores and cylindrical conidia by lacking stipe extensions and vesicles, and by the presence of brown to dark brown, thick-walled setae.

As a later homonym of Mycosphaerella aggregata (Schweinitz) Stevenson the name Mycosphaerella aggregata Carnegie & Keane is illegitimate. The name Mycosphaerella gregaria is proposed as a replacement with similar etymology. The original description is revised following further collections and includes new hosts and cultural characteristics. Originally thought to be a saprophyte, this species may be a primary pathogen of leaves of Eucalyptus.

The occurrence of an undescribed ergot species, Claviceps citrina, on the halophytic chloridoid grass Distichlis spicata, in the Texcoco region of central Mexico, is reported. RAPD and ITS1 sequences of this fungus were compared with other Claviceps species and the morphological uniqueness of the fungus was reinforced. Its sclerotia do not contain any alkaloids of the ergoline type. Germination of sclerotia occurred immediately after placing them on humid sand. After removal of the capitula, the remaining stipe was able to regenerate the capitulum.

A xylanase gene, XYL2, was identified and characterised in Cochliobolus sativus (anamorph Bipolaris sorokiniana), a necrotrophic cereal pathogen that attacks both shoots and roots. The fungus was grown on a xylanase inducing medium containing mineral salts, oat spelt xylan, cellulose, and peptone, RNA was isolated, and a complementary DNA (cDNA) library constructed. The library was screened with a xylanase (XYL1) cDNA clone from the maize pathogen Cochliobolus carbonum. Xylanase cDNA clones, all representing a single gene, were identified. Corresponding genomic DNA was amplified by PCR. Sequencing of the cDNA and the PCR products gave a nucleotide sequence of 2211 bp containing two introns in an open reading frame of 693 bp that codes for a xylanase from glycosyl hydrolase family 11. The most similar sequences to this gene in nucleotide sequence databases are the XYL2 gene of C. carbonum and a xylanase (XYL1) cDNA from a saprophytic fungus, Humicola insolens. Northern blot analysis and reverse transcription PCR (RT-PCR) showed expression of the gene when the fungus was grown on xylan or cellulose, but not when peptone or sucrose was the only carbon source. Expression of XYL2 in inoculated barley seedlings was detected by RT-PCR.