This issue of Mycological Research News points out that Micropeltopsis quinquinecladiopsis belongs in Lichenopeltella, and features a record number of ascomycetes found on a single tree in Papua New Guinea, and the rules of fungal melanins.

This part of Mycological Research includes 14 papers. The first reports the transformation of the endophyte Neotyphoidiumlolii with a green fluorescent protein gene. Molecular systematic studies clarify generic and species concepts in Phaeoacremonium and Phaeomoniella, reassess Neofabraea species, compare species of Hypoxylon and allied genera, clarify the identities of Oidium powdery mildews on tomato, and provide evidence for the reproductive mode in Phellinus nigrolimitatus.

The intracellular peptidases and proteinases of Serpula lacrymans have been investigated, and conidial discharge in Erynia neoaphidis examined.

Developmental studies are reported on apothecia of Pyrenopeziza brassicae and basidiomes in Mycena stylobates, and the germination of ascospores of Monosporascus cannonballus documented. The moss parasites Eocronartium and Jola have been studied in culture to elucidate their life-histories and revealed anamorphs.

Additional fungi have been found in the hypersaline Dead Sea, and seven new neotropical smut species are described.

The following new scientific names are introduced: Aurantiosporium pallidum, Kuntzeomyces ruiziana, Moreaua bulbostylidis, Oidium neolycopersici, Phaeoacremonium mortoniae, Thecaphora amaranthicola, T. smallanthi and Tilletia boliviana spp. nov.; and Lichenopeltella quinquinecladiopsis (syn. Micropeltopsis quinquinecladiopsis) comb. nov.

The green fluorescent protein gene was inserted into isolates of the endophyte Neotyphodium lolii from perennial ryegrass Lolium perenne using the method with polyethylene glycol treatment of protoplasts. The transformation rate was 3·2×10−6 of the original number of protoplasts 5 weeks after the transformation. Freshly isolated mycelium from the host plant was used in the transformation experiments, since difficulties of infecting the host plant with older isolates were observed. Isolates were identified using the universal primed (UP) PCR fingerprinting method.

The transformed mycelium readily expressed the green fluorescent protein gene. Fluorescence was observed in the cytoplasm of the mycelium cells in pure culture, both before infection and after re-isolation from the host plant. The fluorescent hyphae were easily detected in the host plant. No disturbing autofluorescence from the host plant leaf sheaths or from the wild type isolate was observed. The importance of mitotic stability is discussed, based on the transformed phenotype and southern hybridization pattern, since the Neotyphodium lolii transformants lost 1–2 copies of the integrated gene after re-isolation from the host plant without losing their GFP phenotype. This is the first report on transformation of a Neotyphodium sp. with a gfp gene. In the future, the green fluorescent protein may serve as a tool for studying gene expression and protein localization in the endophyte/host plant interaction using heterologous fusion proteins.

Based on ITS and β-tubulin sequence data of 33 isolates, the newly introduced genus, Phaeomoniella was confirmed as being distinct from Phaeoacremonium (Pm.). Phylogeny inferred from DNA sequences and cultural characteristics also confirmed the species status of Pm. aleophilum and Pm. angustius, which were recently reduced to synonymy. Pm. aleophilum has an optimum growth rate at 30 °C and the ability to grow at 35°, whereas Pm. angustius has an optimum growth rate at 25 °C and does not grow at 35°. Furthermore, ITS and β-tubulin sequence data showed Pm. viticola to be indistinguishable from Pm. angustius, while a new species, Pm. mortoniae, could be distinguished from this complex.

Three fungal species responsible for anthracnose canker, perennial canker, and bull's-eye rot of apple have been considered members of the genus Pezicula for a number of years. Recent studies, however, have provided evidence to (re-)classify these species as Neofabraea. There has been a long historical debate regarding the taxonomy of two of these fungi. In Europe, both Neofabraea malicorticis and N. perennans have generally been considered N. malicorticis, while in North America a species distinction has been maintained. Phylogenetic analyses of Neofabraea isolates were based on DNA sequences of the internal transcribed spacer region of nuclear rDNA (38 isolates), the mitochondrial rDNA small subunit (partial; 48 isolates), the β-tubulin gene (partial; 25 isolates), and a combined data set (21 isolates). Our work provides evidence for the existence of four distinct Neofabraea apple pathogens including N. malicorticis, N. perennans, N. alba, and a putative new Neofabraea species that was isolated in both Europe and eastern North America. Our results indicate that the primary Neofabraea species causing tree cankers and bull's-eye rot in North America are N. malicorticis and N. perennans in the west, and N. alba in eastern Canada. N. perennans, N. alba, and the undescribed Neofabraea species were found in Europe but the presence of N. malicorticis was not confirmed by our limited sampling. Inclusion of Rosa spp. in the host range of N. malicorticis is merited.

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.

The aim of this study was to clarify the reproductive mode in the wood-rotting fungus Phellinus nigrolimitatus, using molecular markers and pairing experiments of cultural isolates. A 511 bp segment of the translation elongation factor 1a (EF1a) gene and the internal and intergenic transcribed spacer sequences (ITS and IGS) of the nuclear ribosomal DNA (nrDNA), were amplified by PCR from somatic cultures derived directly from basidiomes and single spore cultures of the fungus. Sequence analyses of the partial EF1a sequence (511 bp) and ITS (732 bp) of single spore cultures gave five and fourteen variable sites, respectively. Five somatic isolates yielded double ITS sequences, which suggested the existence of different ITS types within the same isolate. Restriction digestion of the partial EF1a and the complete ITS and IGS (3·4±0·2 kbp) sequences with HinfI and HaeIII gave restriction fragment length polymorphisms (RFLPs) among the isolates. The sum of fragments exceeded the lengths of the initial PCR product in several somatic isolates, which indicated the existence of EF1a and nrDNA heterotypes within isolates. Segregation of ITS and IGS RFLPs in single spore isolates suggested heterokaryosis in somatic isolates of P. nigrolimitatus. The polymorphic EF1a and ITS restriction sites were used as codominant markers in a test for Hardy–Weinberg expectations. The molecular data suggests a predominant outcrossing reproductive mode in P. nigrolimitatus. Results from cultural pairing experiments with single spore isolates corresponded poorly with the molecular data, suggesting that laboratory pairing experiments do not necessarily provide reliable information of the mode of reproduction in natural populations of fungi.

A world-wide study of the Oidium species causing economic damage on tomato has identified two taxa using classical morphological, scanning electron microscope (SEM) and molecular phylogenetic analyses. The material consisted of a total of 25 tomato powdery mildew isolates and 29 herbarium specimens coming from all continents where tomatoes are grown. A taxon with non-catenate conidia widespread in Europe, Africa, North and South America and Asia was identified as an O. subgen. Pseudoidium species (teleomorph: Erysiphe sect. Erysiphe). Formerly mistaken for O. lycopersicum (or O. lycopersici), it is now recognised as a distinct species, O. neolycopersici sp. nov. A phylogenetic analysis of the internal transcribed spacer (ITS) sequences of the ribosomal DNA (rDNA) indicated that O. neolycopersici is closely related to Erysiphe macleayae, E. aquilegiae and other Pseudoidium species. Only a taxon with catenate conidia was found on Australian specimens. This was identified as a species of O. subgen. Reticuloidium (teleomorph: Golovinomyces sp.). Phylogenetic analysis of the rDNA ITS sequences showed that this species is closely related to O. longipes infecting eggplant. Because it is most likely to be the same species as the original O. lycopersicum, which was actually first described in Australia, this is here neotypified as O. lycopersici.

Serpula lacrymans is a basidiomycete cord-forming wood decay fungus which reallocates nitrogen within an extensive perennial mycelial system in response to spatial discontinuities in external nutrient supply. Intracellular stored protein is mobilised by conversion to amino acids at nutrient-poor sites within a mycelium or when a whole mycelium is starved. Intracellular peptidase and proteinase activities of the mycelium were investigated with the aim of identifying proteases specifically activated in response to a nitrogen demand. Mycelium for enzyme extraction grown as surface mats in static liquid culture was homogenised, and the extract used in assays for proteinase and peptidase with various synthetic peptide substrates conjugated to 4-nitroaniline. Activities against different substrates were characterised with respect to pH, inhibitor sensitivity, requirements for divalent metal ions, isoelectric point, and by changes in activities in starved mycelium. Four different activities were found, comprising two peptidases one of which had metalloprotease characteristics, a serine-type proteinase, and a proteinase active at pH 2.5 which was not affected by any of the inhibitors tried. Both the latter were most active in starved mycelium. Isoelectric focusing showed peaks with activities corresponding to the serine-type proteinase and one of the manganese-activated peptidases. Possible roles for these enzymes in nitrogen reallocation during mycelial foraging are discussed.

The development of apothecia of Pyrenopeziza brassicae (anamorph Cylindrosporium concentricum) on oilseed rape debris and compost malt agar was observed by scanning electron and light microscopy. On oilseed rape debris, apothecia developed directly beneath the epidermis as small globular structures of dense mycelium, which protruded through the epidermis as they increased in size. The apices of erumpent immature apothecia then developed small ostiolar openings which increased in diameter to expose the hymenia of mature apothecia containing dome-shaped asci interspersed with filiform paraphyses. On compost malt agar, hyphae of one mating type grew towards hyphae of the opposite mating type 3–4 d after inoculation of conidia onto agar surfaces. The subsequent development of apothecia on compost malt agar was similar to that on oilseed rape debris.

The patterns of conidial discharge of Erynia neoaphidis were measured from three species of aphids: nettle aphid (Microlophium carnosum), grain aphid (Sitobion avenae) and pea aphid (Acyrthosiphon pisum). The effects of release height, morph of aphid and temperature on the horizontal and vertical discharge of conidia were studied. Numbers of conidia deposited in the dorsal and lateral directions were distributed with distance in truncated bell shaped patterns. Discharge distances ranged from 2 to 11 mm and half the conidia caught travelled further than 5 mm. There was little difference in the horizontal dispersal patterns of conidia when ejected from cadavers of different morphs of the same species, or from different species. The weight of infected aphids showed little correlation with discharge patterns. Values of maximum discharge distance, Dm, were generally between 6 and 9 mm. Temperature had a significant effect on conidia discharged from the dorsal surface of apterous A. pisum cadavers. Dm was greater at 18 °C than at 10 or 25 °. Vertical discharge distances for conidia released from A. pisum cadavers ranged between about 2 and 8 mm. The maximum height to which conidia were projected vertically was not affected by temperature. However, the average height tended to be greater at 18 °. Half of the conidia reached heights of about 3.5 mm while the maximum heights reached were about 8 mm. The initial speeds of conidia were estimated from measured discharge distances. The results suggest that conidia of E. neoaphidis may be ejected at speeds of about 8 ms−1.

The ontogeny of the fruit bodies of Mycena stylobates was studied by light microscopy and scanning electron microscopy (SEM). The ontogeny was divided into two phases: first the primordium with all the structures of the mature fruit body was established, then the primordial stipe elongated rapidly and the exposed hymenium started producing spores immediately. The first detected stage of fruit body formation was an irregularly arranged hyphal structure within the substrate colonized. After the rupture of the surface of the substrate, the primordium established on the surface. Soon a layer of wrapping hyphae was differentiated, which covered the complete primordium. The structures of the stipe and the cap developed synchronously. The developing stipe, cap and basal disc together formed a secondary ring-like cavity, in which the development of the hymenophore took place. The lower side of the cap was covered by a layer of degenerated hyphae. The development of the hymenophore started with a number of small alveolae on the lower side of the cap, which were covered with a hymenophoral palisade. The margins of these alveolae formed the primary lamellae, which in the first stage of their development were covered by a layer of degenerated hyphae. The hymenophoral palisade spread from the developing alveolae to the lamellar edge; the edge of the primary lamellae was forked in the early stages. Secondary lamellae were formed by the down folding of ridges from the lower side of the cap. In contrast to the primary lamellae, they were covered with hymenophoral palisade from the beginning. Spore production started immediately after the elongation of the stipe. These results were compared with other known modes of ontogeny within the Agaricales and some comments on the terminology used for the description of basidiome morphogenesis are made.

The heterobasidiomycete moss parasites Eocronartium and Jola have figured prominently in theories of the evolution of the rust fungi and of the basidiomycetes. Such theories made implicit assumptions about the moss parasites, although very little of their life history was known. This is the first in a series of studies to elucidate the life history of the moss parasites. Polyspore monokaryotic cultures were obtained from basidiospores, and dikaryotic cultures were obtained from hyphae inside the moss host plant. In culture, the moss parasites produced Sporothrix-like anamorphs. The uninucleate conidia germinated by a germ tube, by production of single secondary conidia, or by iterative germination. Conidial-hyphal fusion was observed. A dikaryon was produced by mating single conidial isolates of Eocronartium muscicola, completing a part of its life history. Ultrastructural characteristics of wall break at branching, condensed chromatin during interphase, and simple septal pore morphology from fruiting bodies of Jola spp. and cultured isolates of Jola javensis and E. muscicola were consistent with those of related auricularioid phytoparasitic taxa, the rust fungi, and Pachnocybe ferruginea. Jola and Eocronartium can be grown in axenic culture and are not obligate parasites. In nature, the Sporothrix-like anamorph of these fungi may function in dispersal and mating. The previously unknown anamorph of the moss parasites may be instrumental in our understanding of the origin and evolution of the rust uredinial stage.

Ascospore germlings of Monosporascus cannonballus attached to melon roots were examined with the scanning electron microscope. Results showed that ascospores germinated with one to three germ tubes. Sixty percent of the ascospores germinated with two germ tubes. No germ pores were observed. Instead, germ tubes emerged from a single linear fissure on each ascospore. The largest fissure measured 25 μm in length and 6 μm at its greatest width. Germ tubes of ascospore germlings were firmly anchored to melon roots. However, no structures resembling appressoria were observed. Rather, the tips of germ tubes, upon contact with the epidermis, appeared to have penetrated the epidermis directly. When the germ tube at the point of attachment was dislodged, no infection peg was present at the germ tube tip.

To investigate the spatial and temporal diversity in the fungal community of the Dead Sea, we collected Dead Sea water samples at eight near-shore localities and at different stations offshore over a 1-year period (1999–2000). In addition, depth profiles were sampled at a deep station (304 m) in the centre of the sea. In the course of the study we obtained 476 isolates, comprising 38 species from 19 genera of Oomycota (1), Zygomycota (2), Ascomycota (13), and mitosporic fungi (3). This brings the total number of species recovered from the Dead Sea to 55. Approximately 43% of the isolates belonged to the genera Aspergillus and Eurotium. Most of the species found appeared only in winter. Fungal diversity increased near the outlets of less saline springs near the shore. The species Aspergillus terreus, A. sydowii, A. versicolor, Eurotium herbariorum, Penicillium westlingii, Cladosporium cladosporoides and C. sphaerospermum were isolated consistently and probably form a stable core of the community. The results suggest that a remarkably diverse fungal diversity may be found in the hypersaline Dead Sea waters. To what extent the fungal diversity recovered was present as dormant spores or as vegetative mycelia remains to be determined.

Seven new species of smut fungi from the neotropics are described. The new species are remarkable for their special morphological development, as new representatives of small genera, or by interesting host plants and contribution to our knowledge of neotropic biodiversity. Aurantiosporium pallidum from Bolivia is the third species known in its genus. Its sori develop in the hypertrophied tissue of the rachillae of female and male spikelets. Kuntzeomyces ruizianae from Colombia is the second species known in its genus, characterized by relatively small teliospores. Moreaua bulbostylidis from Bolivia and Venezuela is the first record of a species of this genus (formerly Tolyposporium) on the genus Bulbostylis (Cyperaceae). Thecaphora amaranthicola from Ecuador differs from the known species T. amaranthi on the same host genus by smaller balls of teliospores. Thecaphora smallanthi is the first species of Thecaphora known to include hyaline globose sterile cells in the centre of teliospore balls. Tilletia boliviana is the first species of Tilletia infecting a species of Bromus (Poaceae) and forming warty teliospores. Species of Urocystis on Poaceae are known as agents of stripes on leaves, stems, and inflorescences. Urocystis reinhardii from Bolivia is the first species of this genus with sori in the interior of hypertrophic peduncles of a grass.