Almost 900 species were encountered during a seven year survey of fungal fruit bodies in 13 forests and grasslands plots measuring about 1 ha. The data were comparable to those from a Swiss forest plot monitored for 21 years. Species richness and abundance were almost linearly correlated on a log/log scale. Abundance and yearly frequency of species were tightly correlated. The geographic range of species may also be related to these two parameters. Rare species scored low on abundance, yearly frequency and geographic range. About half of the species were rare, and occurred in only one out of seven years. The two major functional groups of saprotrophic and mycorrhizal species behaved similarly over the years, as in the Swiss study.

Annual variation was high for species richness alone and for richness combined with abundances. The log transformed species richness of plots correlated well with parameters that take the abundances or yearly frequencies of species into account. If other mushroom assemblages show similar regularities, it may be possible to omit abundance counts in future surveys. The species composition of the plots varied strongly over the years. Many species did not reach their maximum abundance in the richest year. Species showed their maximum abundances in different plots in the same year more often than expected by chance. Thus the presence and abundance of species depended on factor(s) other than the general productivity of a year, and years showed (a) different aspect(s) than productivity alone. Long-term surveys are important for understanding the structure of mushroom assemblages and their biodiversity.

Fusarium equiseti and F. pallidoroseum are frequently reported as secondary colonizers of plant tissues. In this study they were isolated from the embryos of weathered cottonseed. Most isolates tested produced equisetin, an antibiotic, when grown on potato dextrose agar, rice, surface-sterilized cottonseed, or autoclaved cottonseed. This is the first report of equisetin from F. pallidoroseum. Equisetin was extracted from cultures of F. equiseti and F. pallidoroseum with acetone and dichloromethane, and partially purified by TLC. Two epimers of equisetin, designated as EQ and epi-EQ, were separated by HPLC. EQ or epi-EQ at 2·5–10 μg ml−1 suppressed germination or inhibited growth of various monocotyledonous and dicotyledonous seed, when the seed were incubated at 30°C under aqueous shake conditions. The two epimers also inhibited the growth of young seedlings and caused necrotic lesions on the roots, cotyledons, and coleoptiles of tested plant seedlings. The results suggest that equisetin may be a pathogenic factor of F. equiseti and F. pallidoroseum on seed and seedling health of cotton and other plants.

The current circumscription of Lasiosphaeria includes taxa with a wide variety of ascomatal walls, ascomatal wall vestitures, and ascospore morphologies and a broad range of putative anamorphs. Despite the complexity of morphological characters in the genus, species within Lasiosphaeria can be arranged into four groups based on ascospore morphology. Taxa which possessed ascospores in each of the four groups were used in phylogenetic analyses of partial nuclear large subunit (LSU) rDNA sequences to test the monophyly of the genus and determine relationships among its species. Lasiosphaeria was found to be highly polyphyletic in that species segregated into seven well-supported monophyletic clades dispersed among several orders. Three new genera, Echinosphaeria, Hilberina, and Immersiella, are erected for three of these clades while the genus Lasiosphaeris is reintroduced for a fourth clade. These data support Ruzenia as a previously established genus and the transfer of Lasiosphaeria raciborskii to Chaetosphaeria. The circumscription of Lasiosphaeria has been considerably narrowed to better reflect a natural classification. These taxonomic changes are additionally supported by a combination of morphological characters which are discussed in relation to the phylogenetic trees.

Botrytis cinerea (anamorph of Botryotinia fuckeliana) is a filamentous ascomycete that causes grey mould especially on grapevine. Based on the presence or the absence of two transposable elements (Boty and Flipper) two sibling sympatric populations named transposa and vacuma have been described. Among the vacuma population, some strains (designated HydR1) were found to be resistant to fenhexamid (a sterol C-4 demethylase inhibitor) and to show an increased sensitivity to 14α-demethylase inhibitors (DMIs). In order to assess whether or not mutations at the target gene level (CYP51) could underlie increased sensitivity to DMIs in HydR1 strains, we cloned the CYP51 gene and determined its DNA sequence in various B. cinerea strains. The gene was highly polymorphic, with mutations detected at 58 positions in the 35 strains analysed. The polymorphisms did not discriminate between transposa and vacuma strains, but did distinguish between HydR1 and non-HydR1 ones. Two expressed mutations were present in all HydR1 strains, namely phenylalanine to leucine at position 15 of the inferred protein, and serine to asparagine at position 105. These data, combined with the existence of morphological differences and somatic incompatibility between HydR1 and non-HydR1 strains, suggest that these two groups comprise distinct genetic entities.

In a continuation of studies started by de Ruiter et al. (1993), all known species of the Mortierella isabellina-group (Micromucor/Umbelopsis clade of O'Donnell et al. 2001) and a few other Mucorales and species of Mortierella were investigated by RFLP (including ITS1, 5.8S, ITS 2 and the 5′ end of the large subunit rDNA gene) and ITS1 sequence analyses. This monophyletic group is unrelated to Mortierella and is only distantly related to the core group of the Mucoraceae. M. longicollis falls outside the Umbelopsis clade. Molecular data resolved two subclades within the M. isabellina-group; however, they are not correlated with any differences in sporangial wall and shape, spore pigmentation and shape, or sporangiophore branching. Therefore we subsume all taxa in one genus, Umbelopsis. The new family Umbelopsidaceae and the new combinations U. isabellina, U. ramanniana, and U. autotrophica are proposed.

Phenotypic and genotypic variability of 18 endophytic Guignardia strains from different host plants (Anacardiaceae: Anacardium giganteum, Myracrodruon urundeuva, Spondias mombin; Apocynaceae: Aspidosperma polyneuron; Ericaceae: Rhododendron sp.; Fabaceae: Bowdichia nitida; Leguminosae: Cassia occidentalis; Rutaceae: Citrus aurantium) growing in different sites in Brazil was assessed by means of morphometric measurements and inter-single-sequence-repeat-anchored polymerase chain reaction (ISSR-PCR) amplifications of the DNA. Morphology of conidia and ascospores and growth rates of the Brazilian isolates corresponded well with those of G. mangiferae. Multiple correspondence analysis (MCA) of the ISSR-PCR data yielded three groups of strains, which did, however, not correspond either to the host or to the geographic origin. The same individual tree hosted genotypically different strains indicating multiple infections. Phylogeny based on ITS1-5.8S-ITS2 sequence analyses confirmed conspecificity of the Brazilian isolates with G. mangiferae (syn. G. endophyllicola).

Macrophomina phaseolina, the causal agent of charcoal root or collar rot, is an important plant pathogen especially in soybean and cotton. Single primers of simple sequence repeats (SSR) or microsatellite markers have been used for the characterization of genetic variability of different populations of M. phaseolina obtained from soybean and cotton grown in India and the USA. Genetic similarity between isolates was calculated, and cluster analysis was used to generate a dendrogram showing relationships between isolates collected from the two hosts. Forty isolates could be clustered into three major groups corresponding to their hosts and geographical region. The wide distribution of microsatellites in M. phaseolina genome was assessed by agarose gel electrophoresis of the PCR products generated by direct amplification of inter SSR regions DNA. This is the first report of the use of microsatellite markers to characterize the charcoal root rot pathogen. The SSR fingerprints (0.25–3.5 kb) generated using DNA from different populations of M. phaseolina of two hosts indicated that these repeats are interspersed within the genome of this pathogen. The variability found within closely related isolates of M. phaseolina indicated that such microsatellites are useful in population studies and represents a step towards identification of potential isolate diagnostic markers specific to soybean and cotton.

The life-cycle of many plant-pathogenic fungi, especially those infecting aerial plant organs, contains several specific developmental stages. If these are sufficiently distinct in their physiology from vegetative hyphal growth, they present potential targets for non-fungitoxic plant protectants. The present review identifies such targets especially in the pre-penetration stages of the infection cycle of Magnaporthe grisea and other fungi infecting from air-borne spores. Examples of non-toxic natural products with activity against spore germination, attachment, appressorium formation, appressorium maturation and penetration of the host surface are given. In contrast, no substances selectively active against in planta growth or sporulation appear to be known. The selective activity of numerous secondary metabolites against specific infection stages without accompanying toxicity against vegetatively growing hyphae indicates a direction for the development of future natural product-derived fungicides which are more easily degraded in the environment and possess fewer non-target effects. Such substances are produced by many saprotrophic and endophytic fungi in pure culture. The paucity of data on the production of biologically active substances in natural situations limits the interpretation of their ecophysiological significance for the producer.

Conserved primers were used in a polymerase chain reaction to amplify the ITS region of the rDNA of 100 Microdochium nivale isolates collected from different turfgrasses in southern Ontario. The profile of the restriction digestion of the amplified ITS region revealed that all the M. nivale isolates analysed belonged to var. nivale. RAPD profiling and RFLP analyses of the IGS regions of rDNA revealed extensive genetic diversity within var. nivale. With RAPD markers, the average similarity coefficient was 66% and the estimate of genotypic diversity was 0·179. Population subdivision analysis showed that 92·2% of the total genetic diversity was found among individuals within populations compared to 7·8% among populations. In dendrograms derived from genetic distances using RAPD and IGS-RFLP markers, there was some evidence for host specialization. Most RAPD markers were shared by individuals from different turfgrasses, and the populations were not highly differentiated. The high level of genotypic diversity detected within populations and the low level of genetic differentiation among populations show that recombination and migration are likely playing important roles in the population biology of M. nivale var. nivale.

Progress in understanding the biology of arbuscular mycorrhizal fungi is hampered by the limited number of species that can be successfully propagated and studied in vitro. We report the establishment of monoxenic cultures of Glomus etunicatum in association with excised Ri T-DNA transformed carrot roots. The fungus can be propagated in vitro using monoxenically formed resting spores and/or colonized root fragments. Modified White's medium buffered with 10 mM MES (pH 6) or MOPSO (pH 6·5) was most optimal for the host root growth as well as for G. etunicatum spore germination and mycorrhiza formation. The number of resting spores formed in vitro correlated positively with the length of roots occupied by arbuscular mycorrhizal structures, including arbuscules and vesicles. Spores first appeared in dual cultures within two weeks of root inoculation. Sporulation was asynchronous and continued until root senescence. Under applied culture conditions, spores achieved mature appearance within 5–7 d after their initiation. Approximately 6% of monoxenic spores were aborted at different stages of their development. Although G. etunicatum spores formed in vitro exhibited general morphological and anatomical similarity to soil-borne inoculum, they were significantly smaller and had thicker spore walls than their soil-borne counterparts. Caution should, therefore, be exercised in utilizing the in vitro system as a model of growth and development of glomalean fungi in soil.

Trichoderma harzianum is a ubiquitously distributed asexual soil fungus that produces a variety of antibiotic compounds. Colonisation of soil inhabited by competing microbiota is facilitated by the antibiotic activity of these compounds. In addition, T. harzianum produces hydrolytic enzymes that degrade the cell wall components of many microorganisms, which can then be used as a source of nutrients. Recently, biotypes of T. harzianum differing morphologically from those originally described by Rifai were isolated on commercial mushroom (Agaricus bisporus) farms. These ‘aggressive’ biotypes cause devastating crop loss on mushroom farms. The aggressive biotype in North America was originally known as ‘Th4’ but has been recently renamed Trichoderma aggressivum f. aggressivum. In contrast, ‘non-aggressive’ biotypes, have no noticeable effect on the crop, are similar to T. harzianum and are commonly found on mushroom farms. The mechanism of disease establishment is unknown. We have identified a metabolite produced by T. aggressivum isolates in vitro that inhibits growth of A. bisporus and other fungi. This antifungal compound is not produced by ‘non-aggressive’ T. harzianum isolates under the culture conditions tested and is identified as 3,4-dihydro-8-hydroxy-3-methylisocoumarin. Another compound was isolated from both liquid culture and infested compost. Although its chemical structure could not be precisely determined, this compound also inhibits A. bisporus growth, is predominant in infested compost and likely has a inhibitory effect on the mycelia present in mushroom compost, resulting in devastating crop loss.

This paper synthesizes the work done on decolourization of bleach plant effluent and synthetic dyes using fungi from marine habitats. Three fungi obtained from marine habitats removed up to 60–95% of Kraft paper mill bleach plant effluent colour. One of the cultures could bring about decolourization of bleach plant effluent at an alkaline pH of 8.2. All the three fungi produced the lignin-modifying enzymes laccase and manganese-dependent peroxidase (MNP) and one of the cultures, Flavodon flavus (isolate 312), also produced lignin peroxidase (LIP) when grown in low nitrogen medium. Production of these enzymes increased in the presence of paper mill effluent in all the three isolates. Isolate Flavodon flavus (312) decolourized synthetic dyes like Azure B, Brilliant green, Congo red, Crystal violet, Poly-R, Poly-B and Remazol Brilliant Blue R in low nitrogen medium prepared with distilled as well as with artificial sea water. Decolourization of dyes was also observed in a rich nutrient medium like malt extract broth. About 60–90% of decolourization was achieved in these media within 3–10 days. Visible spectra of most of the dyes incubated with this culture indicated degradation at the end of the experiment. Manganese-dependent peroxidase enzyme was always produced in cultures of 312 containing dyes and in some cases, a direct correlation existed between % decolourization and MNP activity, suggesting its involvement in this process. This culture with its capacity to degrade paper mill effluent and synthetic dyes appears to be a potential candidate for bioremediation of coloured industrial effluents.

The effect of simulated rain on the persistence of oil and water formulations of conidia of the entomogenous fungus Metarhizium anisopliae when applied to oilseed rape foliage was investigated, using third instar larvae of the mustard beetle (Phaedon cochleariae) as the target host. Rain significantly (P < 0.01) reduced the susceptibility of the beetle larvae to M. anisopliae but the amount of inoculum removed was influenced by the formulation. Larvae exposed to plants treated with conidia formulated in aqueous Tween, Shellsol T, or sunflower oil/Shellsol T resulted in 55, 82.5 and 72.5%, mortality, respectively. The mortality for these respective formulations was reduced by 42, 57 and 51% if the plants were exposed for 1 h to simulated rain. Laboratory and field studies showed that more inoculum collected beneath plants sprayed with conidia formulated in Shellsol T or aqueous Tween than in the more viscous sunflower/Shellsol T mixture. Mortality studies on leaves taken from field plots suggested that conidia on leaf surfaces could be replenished by repeated application. The number of conidia isolated from field plots was greater where inoculum was applied bi-weekly than once weekly.

Although Eutypa lata is the main agent of Eutypa dieback of grapevine, another species of Eutypa has been isolated from diseased grapevines in Northern California. Stromata of this recently discovered Eutypa were also collected from Acer macrophyllum, A. negundo, and Fraxinus latifolia in the vicinity of vineyards, and appeared commonly on Umbellularia californica in some mixed-evergreen forests of Napa and Sonoma counties. This second species of Eutypa was distinguished from E. lata because of the sulcate ostiole of the perithecium and smaller ascospores. A morphological comparison with type specimens revealed identical features between the Californian isolates and E. leptoplaca sensu Rappaz (1987). This identification was confirmed through phylogenetic analyses of Eutypa spp. based on the complete sequence of the internal transcribed spacer (ITS) of the rDNA and partial sequence of the β-tubulin gene. These analyses also separated collections of E. maura, E. sparsa, E. lejoplaca, E. tetragona, E. leptoplaca and E. lata, confirming the previously proposed species concepts. The pathogenicity of E. leptoplaca on grapevine was established using isolates collected from Vitis vinifera, U. californica, and A. macrophyllum. The importance of E. leptoplaca in relation to Eutypa dieback and its role as a necrotrophic pathogen are discussed.

Pycnidial fungi belonging to the genus Ampelomyces are the most common natural antagonists of powdery mildews worldwide. During a study of the interactions between apple powdery mildew (Podosphaera leucotricha) and Ampelomyces mycoparasites, 52 new Ampelomyces isolates were obtained from P. leucotricha and, in addition, 13 new isolates from other species of the Erysiphaceae in four European countries. Their genetic diversity was screened using single-stranded conformation polymorphism (SSCP) analysis of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA). For comparison, 24 isolates obtained from genetic resource collections or other sources were included in this study. Based on the ITS-SSCP patterns, the isolates were placed in eight groups. The isolates belonged to two types based on their growth in culture. The faster-growing and the slower-growing isolates were included in different SSCP groups. A phylogenetic analysis of the ITS sequences of representatives of these groups confirmed the results obtained with the SSCP method, and showed that the faster-growing isolates do not belong to Ampelomyces as suggested by earlier studies. All the isolates from P. leucotricha fell into a distinct SSCP group of genetically homogeneous isolates. This suggests that Ampelomyces mycoparasites which occur in apple powdery mildew are slightly different from the other Ampelomyces groups which contain mycoparasites from various powdery mildew species. This may be because the main growth period of Ampelomyces mycoparasites in apple powdery mildew is isolated in time from that of Ampelomyces isolates that occur in other species of the Erysiphaceae. P. leucotricha starts its life-cycle early in the season, usually in March–April, while most powdery mildews are active in the same environments only late in the year.

This study investigated the morphology and infectivity of fine endophyte in a mediterranean environment. Five morphological groups of fine endophyte differing in up to 11 characters were present within two pot cultures of fine endophyte from south- western Australia. Occurrence and abundance of a particular type of fine endophyte were not affected by the application of calcium carbonate. It appeared that there may be more than one fungus included among the two sources of fine endophyte, however it was not possible to separate the fungi. A second experiment demonstrated that infective propagules of fine endophyte can be very small (<38 μm) and fresh roots acted as a potential source of infective propagules. There was no infectivity of fine endophyte in the soil at the end of the growing season but it reoccurred in similar quantity in the next growing season.

Sequence analysis of the ITS region of rDNA was used to investigate the level of genetic diversity occurring within Hyaloperonospora parasitica, and to show the relationship between phylogenies of these fungi and their hosts (Brassicaceae). Maximum parsimony and neighbour-joining analyses were performed using sequences from 32 isolates of Hyaloperonospora and Perofascia, which infect core brassicaceous hosts. For comparison, five isolates of Peronospora were also studied. The constructed phylogenetic trees showed trichotomy, showing that Hyaloperonospora, Perofascia, and Peronospora have different evolutionary histories. Although isolates from Peronospora and Perofascia clearly formed respective clades, the Hyaloperonospora group allowed separation of the isolates into four distinct clades, which shared significantly low sequence similarities. We suggest that H. parasitica infecting brassicaceous hosts should be divided into a number of distinct species. The comparison of the phylogeny of H. parasitica and that of the Brassicaceae suggests that this fungus is closely related with tribes Arabidae and Brassiceae within this host family, illustrating the potential of downy mildews for co-evolution with their hosts.

We evaluated the possibilities of exploitation of several mycoparasitic fungi in the biocontrol of cucurbit powdery mildew (Sphaerotheca fusca) in melon greenhouses. To simplify the screening, an in vitro biological control test on detached leaves of melon has been used and a detailed microscopic analysis of the interactions between mycoparasites and S. fusca conducted. In this context, the effect of mycoparasitic fungi on the formation of infection structures of S. fusca such as haustoria, conidia and conidiospores has been quantified. On the basis of the microscopic data, effect of mycoparasites on severity and incidence of cucurbit powdery mildew has been discussed. Our results show that, under controlled environmental conditions, Acremonium alternatum, Ampelomyces quisqualis and especially Lecanicillium lecanii, when applied in early stages of infection, were able to significantly reduce cucurbit powdery mildew symptoms and S. fusca development on melon leaves. These results indicate that these mycoparasites are promising candidates for the biocontrol of cucurbit powdery mildew in melon greenhouses.

Phylogenetic analyses of partial sequences (1002 basepairs) from the nuclear encoded small subunit ribosomal RNA (18S rRNA) were performed to determine the phylogenetic relationships of the ascostromatic fungi, Botryosphaeria, Guignardia, Leptosphaerulina, Didymella, Venturia, Sporormia, and Sporormiella. Our studies confirm the concept that the subclass Loculoascomycetidae is paraphyletic if Chaetothyriales is included, as this order and the plectomycetes constitute a monophyletic clade that is highly supported in our and some previous phylogenetic analyses. In general, taxa that lack pseudoparaphyses (Dothideales) or have reduced hamathecial tissue, such as Botryosphaeria and Guignardia (sensu Sivanesan), are a sister group to the Pleosporales, the most derived group, whose members have pseudoparaphyses. The two species of Leptosphaerulina, which also lack pseudoparaphyses, clustered within the Pleosporales, rather than within the Dothideales, with high confidence from bootstrap values. Didymella nested within the Pleosporales with a high degree of confidence, and is the sister group to Leptosphaerulina. The taxonomic placement of Venturia and Sporormiella corresponds to the Pleosporales.

Phylogeny and biogeography of Lentinula, which includes cultivated shiitake mushrooms, were investigated using parsimony analyses of an expanded nuclear ribosomal DNA dataset. Lentinula occurs in the New World as well as Asia and Australasia. The Asian–Australasian Lentinula populations appear to form a clade, but species limits within this group are controversial. We refer to the entire Asian–Australasian Lentinula clade as shiitake. Thirty-seven wild-collected isolates of shiitake were examined, representing Australia, Borneo, China, Japan, Korea, Nepal, New Zealand, Papua New Guinea (PNG), Tasmania and Thailand. Five isolates of the New World species, L. boryana, were included for rooting purposes. Levels of sequence divergence between North and Central American L. boryana isolates are higher than those between the most divergent shiitake isolates. In shiitake, five independent lineages of rDNA were identified, which we call groups I–V, but relationships among these lineages are not well resolved. Group I includes populations from northeast Asia to the South Pacific. Group II includes populations from PNG, Australia and Tasmania. Group III is limited to New Zealand. Group IV is from PNG. Finally, group V is from eastern China and Nepal. The distribution of rDNA lineages suggests a complex biogeographic history. Although many areas remain unsampled, our results suggest that certain areas have particularly high levels of diversity and should be targeted for further study and conservation.