This issue of Mycological Research News features: New BMS website in action; How old are the fungi? Fourteen research papers are included. Molecular and experimental taxonomy papers address Psilocybe sect. Psilocybe, the affinities of Lepraria and Leproloma, variation in the bipolar Xanthoria elegans, the early detection of Tilletia tritici in wheat, repeated ITS1 sequences in a Nodulisporium, the cause of Cryphonectria canker on Tibouchina (Cryphonectria parasitica), variation in Beauveria bassiana isolates from different hosts (including the Coffee Berry Borer) and regions. Mating tests have been carried out to clarify the mating types in Ganoderma boninense, and to test species concepts in Polyporus s. str. species from South America.

The melanin biosynthesis pathway of various sapstain fungi has been identified, and protocols for the cryopreservation of Cantharellus cibarius developed.

Ecological papers examine the endophytes of Fagus sylvatica bark in the Apenines, the growth and development of entomogenous Alternaria alternata, fungi on moose dung from different habitats in Sweden, and the location of liquid water within two soil lichens.

No new scientific names are introduced in this part of Mycological Research.

The species of Psilocybe sect. Psilocybe, formerly classified in the genus Deconica, were investigated using morphology, mating behaviour and RAPD analysis. Psilocybe inquilinus and P. crobula do not seem to be closely related. Based on the morphology, two varieties could be accommodated in P. subviscida, namely as vars. subviscida and velata. The mating group of P. montana is characterized by rather thick-walled, dark spores with a fairly large germ-pore. Putative representatives of P. muscorum and P. physaloides freely interbreed with typical P. montana, and, consequently, these taxa are considered to represent one variable species. The ex-type strain of P. chionophila did not mate with isolates of P. montana. One collection of P. chionophila from a lowland habitat, morphologically resembling P. montana, was found to be interfertile with the ex-type strain of P. chionophila, but not with P. montana. We identified several collections as P. magica, which is morphologically similar to P. schoeneti. Mating studies showed that these specimen belong to the same biological species, but failed to mate with the ex-type of P. schoeneti.

The phylogenetic position of members of the entirely asexually reproducing lichen-forming genera Lepraria and Leproloma was investigated using sequence data from the ITS1-5.8S-ITS2 and small subunit (SSU) nuclear ribosomal DNA. Phylogenetic reconstructions were carried out using a likelihood-based Bayesian Markov chain Monte Carlo (MCMC) tree sampling technique and the unweighted least squares optimality criterion, the latter based on maximum likelihood distances obtained via an alignment-free distance estimation technique. The results indicate that most species currently referred to Lepraria and Leproloma form a single monophyletic group. This monophyletic group is the sister group to Stereocaulon and Muhria and belongs in the Stereocaulaceae (Lecanorales, Lecanoromycetes, Ascomycota). This indicates that an ancestor of Lepraria switched from a sexual to an asexual mode of dispersal. Subsequent speciation must have taken place in the absence of sexual processes, which contradicts the view of asexual taxa as ‘evolutionary dead ends’. Leproloma is polyphyletic and nested within Lepraria. Lepraria flavescens is a Lecanora, probably belonging in subgenus Glaucomaria. Lepraria lesdainii and L. obtusatica are unrelated to each other and to other species currently referred to Lepraria or Leproloma. Leprocaulon and Crocynia are distantly related to the core group of Lepraria and Leproloma.

Genetic and physiological diversity was assessed in isolates of the lichen-forming fungus Xanthoria elegans originating from a range of geographic localities and climatic regimes including polar and temperate regions. In vitro cultures were established and isolates examined by production of multi-locus RAPD markers and by sequence comparison of the ITS region of the rDNA tandem repeat unit. Both molecular techniques revealed significant variability. Phylogenetic analysis of RAPD profiles (226 markers) clustered isolates from particular locations together and Mantel's test showed a correlation between genetic divergence and extent of geographic separation. Phylogenies based on ITS sequence divergence were not well supported and did not correlate with the geographic origin of samples, suggesting a single locus approach to be less informative than RAPD data in a study of this nature. Physiological investigations also revealed significant variability between isolates with different geographic origins. Temperature had a significant effect on relative growth rate (RGR) such that X. elegans originating from sites with lower mean annual temperatures had significantly higher RGRs at all test temperatures between 2 ° and 18 °C. Enhanced metabolic activity might be an adaptation for growth in colder climates. The results demonstrate high genetic diversity within this morphologically variable species in response to geographic and environmental factors, and are discussed in relation to data obtained for non-lichenised fungal species and the possibility of cryptic speciation.

A method for the early detection and diagnosis of common bunt, caused by Tilletia tritici, has been developed. The technique is based on a nested PCR amplification of DNA from T. tritici in infected plant inflorescence tissue at the stage of internode elongation. Primers amplifying a 212 bp fragment in T. tritici were designed on the basis of sequences in the ITS2 region in the ribosomal RNA genes. Disease diagnosis obtained with nested PCR amplification correlated well with the conventional disease diagnosis, which involves manual observation of disease symptoms at maturity. With the present method, wheat varieties can be screened for resistance towards common bunt at the first node stage. There was no variation between the ITS2 region from T. tritici, T. laevis, T. controversa, and T. bromi and the primers are therefore not species specific. The PCR diagnosis method has only been validated for common bunt caused by T. tritici, but it is likely also to be applicable to T. laevis and T. controversa.

A Variable Number of Tandem Repeats (VNTR) polymorphism has been identified in the ITS1 region of the rDNA in 13 strains belonging to a species of Nodulisporium producing the novel indole diterpene nodulisporic acid. The number of tandem repeats found in these isolates varies from 2, 4 or 5 repetitions, and produces three length morphs in the ITS1 rDNA, spanning a length difference of 217, 308 and 352 nucleotides. Repeated units are rather variable, and they encompass three tandem repeated motifs of 11 nucleotides. In all the individuals, the variants are positioned in the same physical order, following the distribution of the most parsimonious tree. This suggests that this sequence have been enlarged by slipped-strand mispairing occurrences.

Cryphonectria cubensis is an important canker pathogen of plantation Eucalyptus spp. in tropical and sub-tropical areas of the world, including South Africa. It is best known on Eucalyptus spp., but it also occurs on Syzygium aromaticum (clove). In 1998, C. cubensis was found to cause cankers on the non-myrtaceous hosts Tibouchina urvilleana and T. lepidota in Colombia. In this study, we report on a similar canker disease that has recently been found in South Africa on T. granulosa, commonly grown as an ornamental tree. The identity of the pathogen was determined through morphological comparisons and phylogenetic analyses of ITS and β-tubulin gene sequences. The pathogenicity of the fungus was also tested on T. granulosa and E. grandis. Morphological as well as DNA sequence comparisons showed that the fungus on T. granulosa is the same as C. cubensis occurring on Eucalyptus spp. in South Africa. Pathogenicity tests on T. granulosa and E. grandis clones showed that the fungus from T. granulosa is able to cause cankers on both hosts.

Genetic variability of 95 isolates of Beauveria bassiana from different geographical regions and hosts was assessed using RAPDs and ITS-RFLP molecular markers in order to characterize its genetic variability. Clusterings were obtained using UPGMA and Principal Coordinates Analysis. Four groups were identified with RAPDs. A 930 bp fragment was amplified with primers ITS1 and PN16, and polymorphisms were observed with AluI and MspI, generating two main clusters. Results indicate low genetic variability present in the Colombian B. bassiana population, no association to host type or geographic locality, and a population structure that is not clonal. This characterization sets criteria for determination of bio-insecticide potentiality, introduction of foreign strains and improvement of entomopathogens within an IPM program.

Ganoderma boninense was determined to be heterothallic and tetrapolar with multiple alleles at both mating type loci. Mycelial interactions amongst siblings were assessed to determine if closely related isolates of G. boninense exhibited vegetative incompatibility. Interactions amongst monokaryotic and dikaryotic mycelia generally grouped into four categories ranging from strong antagonism to the absence of antagonism. Pairing of dikaryotic mycelia always resulted in antagonism and the degree of antagonism generally correlated with the relatedness of the isolates. Genetically different dikaryons therefore constitute discrete individuals. This is the first report of such interactions in G. boninense and these findings provide the basis for further studies on natural populations of this species.

Six species of the genus Polyporus s. str. from which monosporic cultures were obtained, were studied to investigate their sexual compatibility. All species tested have an heterothallic multifactorial tetrapolar system. Results of intraspecific matings indicated a certain reproductive isolation of the Argentine and Brazilian collections of P. tenuiculus. Interspecific matings confirmed the separation of the species P. ciliatus and P. tucumanensis, as well as P. dictyopus and P. melanopus. Di-mon matings allowed the separation of P. tenuiculus from P. philippinensis.

The presence of the 1,8-dihydroxynaphthalene (DHN) melanin biosynthesis pathway was demonstrated in several sapstain fungi including Ceratocystis and Ophiostoma, using both chemical inhibitors and molecular techniques. The inhibitor compounds tricyclazole and carpropamid effectively reduced pigmentation at low concentrations in all tested fungal species, but also lead to growth inhibition at higher concentrations. The inhibitor cerulenin prevented fungal growth in all tested fungi at all tested concentrations, likely due to its inhibitory effect on another enzyme, the metabolically critical fatty acid synthase. Partial DNA sequences for the gene encoding scytalone dehydratase (SD) were obtained from species of Ceratocystis and Ophiostoma and found to have homology with known respective DHN-SD gene sequences. Sequence analysis of the partial SD amino acid sequences showed greater than 80% similarity among the sapstain species, and corresponded well with known phylogenies of sapstain fungi based on rDNA sequences. Aside from the work carried out on the isolate O. floccosum 387N, this is the first known documentation of the melanin pigmentation pathway used by species of the sapstain fungi Ceratocystis, Leptographium and Ophiostoma. Furthermore, since no fungus has ever been found, to our knowledge, to have more than one melanin synthesis pathway, we can state that these species are likely only to use the DHN pathway for melanin production.

Cryopreservation of ectomycorrhizal fungi is often difficult, but needed for patent deposits and for enabling experiments on model strains with constant characteristics. In this study we report a protocol for successful cryopreservation of the ectomycorrhizal mushroom Cantharellus cibarius, together with a brief summary of some unsuccessful attempts. A slow freezing rate (0.3°C min−1) of agar cubes carrying the mycelium, and a gentle addition of the cryoprotectants sorbitol and DMSO were important features. This protocol was successfully repeated by ATCC who used it to deposit a patented strain of C. cibarius, and therefore the protocol should also be tested for other ectomycorrhizal basidiomycetes.

Thirty 3–4 yr-old twigs were collected at each of three sampling dates between May 1995 and May 1996 from each of ten approximately 120 yr-old European beech trees in the Tuscan-Emilian Apennines to examine the bark for the presence of endophytic fungi. Five trees with low crown transparency ([les ]5%) and five trees with high crown transparency ([ges ]25%) were compared. Almost all of the 900 examined bark samples were colonized by endophytic fungi. More than 30% of the twigs appeared to host three or more fungal species. Forty-four endophyte species were detected. An Aposphaeria and a Cryptosporiopsis species, Botryosphaeria quercuum, Discula umbrinella and Neohendersonia kickxii occurred most frequently in all of the three samplings and seemed to play a dominant role as endophytes in beech bark. Significant differences in endophyte assemblages between trees with low and trees with high crown transparency could be detected only with respect to the Aposphaeria species. The colonization of tissues by this fungus, possibly a weak pathogen, was predominant on trees with high crown transparency.

The entomopathogenic fungus Alternaria alternata grew equally well on potato dextrose agar (PDA) and also on potato extract agar (PEA). The latter was excellent for sporulation. The fungus grew at a wide temperature range of 15–35 °C with an optimum at 25 °. On PDA sporulation was much reduced as compared to that on PEA. The spores of the fungus germinated only at 100% relative humidity (rh). Darkness significantly increased the germination percentages in the early stages of germination at all temperatures tested. Differences in germination percentages between light and dark conditions were mostly levelled 24 h post incubation. The germ tubes formed distinct appressoria, both on glass slides and on the aphid host Anuraphis nerii, on which appressoria were formed on all parts of the insect bodies, even on the hardest ones, such as the head and thorax. Most appressoria were single globose and formed terminally on hyphal branches. Few were subapical and still fewer compound and multilobed. Detachment of appressoria from their original position revealed an exceedingly smoothened appression area, around which a mucilagenous material was secreted, apparently cementing them onto the host epicuticle. Optimum temperature for appressorium formation was 20–25 °. At these temperatures more than 80% of the emerging germ tubes formed appressoria. The factors affecting spore germination and appressorium formation are discussed in relation to the prevailing environmental conditions in the mediterranean area as well as their role in influencing the spread and activities of the pathogen and the infection of aphids under field conditions.

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.

The effects of the structural features of the thallus and the lichen–substratum interface on soil water infiltration were investigated using low temperature scanning electron microscopy and scanning electron microscopy with backscattered electron imaging. Two morphologically different lichen species were examined, the crustose Diploschistes diacapsis and the squamulose Squamarina lentigera. Water distribution within lichen-forming soil crusts explains the results of earlier infiltration studies. Several anatomical features of the upper cortex gave rise to similar impervious characteristics of the thallus. The fungal material comprising the lichen thallus showed significant hydrophobic features and only when in contact with calcium oxalate crystals or minerals in the soil was this material able to retain water. The lichen–substratum interface appears to play an important role in the hydric strategy of these lichens which in turn affects the penetration of water into the soil.