Mycological Research News features: In this issue; Molecules vs morphology; The name Fusarium moniliforme should no longer be used; and Differential use of Termitomyces by termites.

Molecular methods are used in this issue to investigate the: phylogeography of Hyphoderma setigerum, Mycosphaerellaceae on Proteaceae, Xerocomus chrysenteron complex, identification of endophytes from Pinus tabulaeformis, root-inhabiting Cryptosporiopsis spp., discrimination of Pisolithus spp., variation in Pyrenochaeta lycopersici, Penicillium nalgiovense pencillin production impaired strains, and laccase genes in Trametes sp. Sexual reactions in Zygomycota are shown to be mediated by trisporoid compounds.

The use of particle filtration to study the diversity of rainforest leaf litter fungi is applied to Neolitsea dealbata, an unclamped Lentinellus has been rediscovered (and a key to allied European species provided), and the first fossil clavarioid fungus is described from Cretaceous amber.

The following new scientific names are introduced: Palaeoclavariaceae fam. nov.; Palaeoclavaria gen. nov.; Cryptosporiopsis rhizophila, P. burmites, Xerocomus cisalpinus spp. nov.; Lentinellus herbarum (syn. Lentinus flabelliformis var. herbarum), Mycosphaerella concentrica (syn. Gibbelina concentrica), M. fibrillosa (syn. Teratosphaeria fibrillosa), M. microspora (syn. T. microspora), M. proteae-arboreae (syn. T. proteae-arboreae), X. fennicus (syn. Boletellus fennicus) combs. nov. A lectotype is designated for the name Thelephora setigera (i.e. Hyphoderma setigerum), and a neotype for Lentinus flabelliformis var. herbarum (i.e. Lentinellus herbarum).

Previous studies of morphological variation in the homobasidiomycete Hyphoderma setigerum have lead to suspicions of a species complex. This study explores variation in DNA sequences from the nuclear ribosomal ITS region of 45 specimens from America, Asia, and Europe in a phylogeographic context. Based on molecular analysis, morphological studies, and crossing tests, nine preliminary taxa are shown to exist inside the species complex, and the two previously described segregate species H. subsetigerum and H. nudicephalum are confirmed. The molecular analysis shows evidence of allopatric differentiation over intercontinental distances. Only one of the nine well-supported clades has a geographic distribution spanning more than one continent, probably indicating the importance of vicariance in the evolution of this species complex. The basionym of H. setigerum, Thelephora setigera, is neotypified to fix the application of that name.

Many plant pathogenic foliicolous fungi recorded on Proteaceae hosts in South Africa are reminiscent of members of the Mycosphaerellaceae and their anamorphs. However, these fungi are often unusual and specific to the Proteaceae, and have proved difficult to classify in the past. To address this issue, a phylogenetic analysis of the ITS-1, 5.8S and ITS-2 DNA sequence data was performed to determine relationships between members of the Mycosphaerellaceae and some taxa apparently unique to the Proteaceae. Results from this study have confirmed Batcheloromyces to be affiliated to the Mycosphaerellaceae. Within Mycosphaerella, Batcheloromyces was also shown to be distinct from Stigmina, which clustered separately from Pseudocercospora. The separation between Mycosphaerella and Teratosphaeria (anamorph Trimmatostroma) was shown to be artificial, which was further supported by several Trimmatostroma species clustering within Mycosphaerella. From these data it is concluded that Teratosphaeria should be reduced to synonymy under Mycosphaerella, and that Trimmatostroma represents yet another additional anamorph of Mycosphaerella.

Species delimitation is still controversial in the Xerocomus chrysenteron complex. We have therefore established comprehensible and reliable species concepts based on statistical evaluation of morphological and ecological characters. We examined many collections from different geographical regions and different developmental stages within collections. Quantitative micromorphological characters (basidiospores, pileipellis end cells) were measured in statistically relevant numbers. The same material was used to generate 24 rDNA-LSU sequences, and the results of phylogenetic analyses clearly confirmed our species concepts: spore size and ornamentation, length of the pileipellis end cells and ‘pruinatus-hyphae’ are most valuable characters for the delimitation of species in this complex. Molecular data demonstrated that the X. chrysenteron complex is a monophyletic group. All the examined species (X. chrysenteron, X. cisalpinus, X. pruinatus, X. ripariellus, X. dryophilus, X. fennicus, X. porosporus, and X. rubellus) represent independent lineages. The faintly striate spores, a key character characterising species of section Striatulispori, probably evolved independently. In addition, the ‘pruinatus-hyphae’ have multiple origins, and truncate spore apices are derived at least twice. Xerocomus cisalpinus sp. nov. is characterised by striate spores, the presence of ‘pruinatus-hyphae’ and a pileipellis strongly reminiscent of X. chrysenteron. For reasons of discussion, microscopical data are presented on Boletellus episcopalis for the first time. Xerocomus fennicus (Boletellus) comb. nov. is proposed. We provide descriptions to all included taxa. Our results once more demonstrate that reliably identified and characterised voucher collections are the basic requirement for meaningful phylogenetic studies.

Sterile mycelia isolated from Pinus tabulaeformis were grouped into white morphotype strains based on cultural characteristics. Eighteen of the isolates were randomly selected and identified to various taxonomic levels based on nuclear ribosomal DNA (nrDNA) sequence analysis. The 5.8S gene and flanking internal transcribed spacer (ITS1 and ITS2) regions of nrDNA were amplified and sequenced. Phylogenetic analysis of the 5.8S gene sequences indicated that the white morphotype strains were Ascomycota. Further identification was achieved by means of sequence similarity comparison and phylogenetic analysis of the ITS regions. Results showed that strains WMS9 and WMS10 were Lophodermium species (Rhytismataceae), while strains WMS11, WMS13 and WMS18 were species of Rhytismataceae. Strains WMS2, WMS3, WMS4, WMS5 and WMS6 were identified to Rosellinia, strain WMS1 to Entoleuca, and strain WMS14 to Nemania (Xylariaceae). Strains WMS7, WMS8, WMS12, WMS15, WMS16 and WMS17 were xylariaceous species. The potential of using DNA sequence analysis in the identification of endophytic fungi is discussed.

Three Cryptosporiopsis species have thus far been isolated from roots of woody plants. A fourth species, which was recently isolated from roots of Calluna vulgaris, Erica tetralix, Vaccinium vitis-idaea, and V. myrtillus in The Netherlands, is described here as new. Sporulation on the natural substratum has not been observed and the morphological description of this fungus is therefore based on characters expressed on oatmeal and malt extract agars. The phenotypic characters indicated a close relationship with the other root-inhabiting species of Cryptosporiopsis and species of the associated teleomorph genus Pezicula. This relationship was confirmed by phylogenetic analyses using sequence data of the 5.8S nuclear rDNA and flanking internal transcribed spacers. In order to facilitate recognition of this possibly under-recognized category of root inhabitants, a key to the root-inhabiting Cryptosporiopsis species based on characters in vitro is given.

Inter-simple sequence repeat PCR (ISSR-PCR) was used to develop markers for simple sequence repeat-rich (SSR) regions for investigation of genetic relatedness of Pisolithus isolates collected from eastern mainland Australia. Primers were designed to amplify ten SSR-rich regions and these were used to screen 14 Pisolithus isolates. Two amplified loci showed size polymorphisms among the isolates (regarded as polymorphic), two were monomorphic for all isolates, while the remainder amplified alleles for only some isolates. UPGMA analysis of the alleles for each isolate at each locus together with ITS-RFLP analysis, separated the isolates into two groups. These two groups appear to correspond to isolates that ITS sequence data have previously separated as P. albus and P. microcarpus.

Corky root of tomato caused by Pyrenochaeta lycopersici is a disease of concern in Italy and for many tomato growing areas in the world. Isolates of the fungus were characterized at both the physiological and molecular level. The optimal in vitro growth temperature for all isolates was 23 °C. All Italian isolates of P. lycopersici showed similar RAPD and esterase banding patterns. No relevant polymorphisms were detected after enzymatic digestion of PCR-amplified ITS and IGS regions. The overall results indicate a low degree of genetic variability within a collection of 43 Italian isolates. These data are of interest in breeding programs for resistance against corky root of tomato and they provide useful information for the development of molecular diagnostic tools for the rapid identification and detection of P. lycopersici.

The presence of some fungi on a variety of food products, like cheeses or cured meat products, is beneficial for the ripening of the product and for the development of specific flavour features. The utilization of these fungi as starters, which are inoculated normally as asexual spores on the food products at the beginning of the ripening process, is becoming a usual procedure in the food industry. The starter culture also prevents undesirable fungi or bacteria from growing on the product. Penicillium nalgiovense is the most frequently used starter for cured and fermented meat products, but the fact that this fungus can secrete penicillin to the meat product makes it important to get strains unable to synthesize this antibiotic. In this work we report that P. nalgiovense strains impaired in penicillin production can be obtained by disruption of the pcbAB gene (the first gene of the penicillin biosynthetic pathway). When applied as starter on cecina (a salted, smoke-cured beef meat product from the region of León, Spain), the pcbAB-disrupted strain showed no differences with respect to the parental penicillin-producing strain in its ability to colonize the meat pieces and to control their normal mycoflora. Both strains exerted a similar control on the presence of bacteria in cecina. A similar proportion of penicillin-sensitive and penicillin-resistant bacteria were isolated from pieces inoculated with the penicillin-producing or the non-producing P. nalgiovense strains. The decrease of the bacterial population on the surface of cecina seems to be due to the higher competition for nutrients as a consequence of the inoculation and development of the P. nalgiovense mycelium and not due to the production of penicillin by this fungus. Penicillin production was less affected than growth in a solid medium with high NaCl concentrations; this suggests that the high salt concentration present in cecina is not a limiting factor for penicillin production by P. nalgiovense.

The strain Trametes sp. I-62 (CECT 20197) is a white-rot fungus with great potential for biotechnological applications in the fields of industrial waste water decolorization and clean up. Three laccase genes: lcc1, lcc2 and lcc3 have been cloned and sequenced from this basidiomycete. In this work, the coding regions of the corresponding cDNAs have been synthesized, cloned, and sequenced. They are 1563, 1563 and 1575 bp in length, respectively. Former putative intron/exon structures from genomic DNA are fully confirmed by match analysis with our cDNA sequences. Using Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR–RFLP) analysis, an additional laccase cDNA was also identified, corresponding to a new gene, lcc1A, which displayed 99.6% identity with lcc1 at protein level. Such high similarity between lcc1 and lcc1A sequences, and the comparison with reports from other basidiomycete laccases, suggest that in this strain these two genes are allelic variants.

Several species of Mortierella (Mortierellales, Zygomycota) were examined for substances regulating their sexual reactions. Compounds isolated from both mated and single growing Mortierella strains were purified by thin layer chromatography. Some of these compounds showed UV absorbance-characteristics similar to those of trisporoids, a group of compounds involved in sexual regulation in Mucorales. A compound with a 4-dihydromethyltrisporate-like absorbance spectrum was detected. To test for the interspecific sexual responses typically induced by trisporoids, the compounds extracted from Mortierella spp. were tested against the Mucorales Mucor mucedo and Phycomyces blakesleeanus and were found to induce sexual reactions in both tester strains. A gene encoding 4-dihydromethyltrisporate dehydrogenase was identified in several Mortierella species and the activity of the gene product was shown using a histochemical assay. We suggest that the regulation of sexual processes by trisporoids is common to both Mucorales and Mortierellales and may be more widespread within the Zygomycota.

Fungal species richness and abundance were assessed in leaf litter of the Australian rainforest tree Neolitsea dealbata (Lauraceae) using particle filtration. Results were comparable to the species richness and abundance reported in previous studies of tropical leaf litter microfungi. Eight leaf samples yielded 1365 strains. In an assessment of the effect of surface treatments, 736 strains in 112 morphotaxa were isolated, while 639 strains in 141 morphotaxa were recovered to assess the effect of surface treatment. Isolation rates in airdried leaves stored at room temperature for four weeks declined linearly, while the number of morphotaxa remained essentially constant for the first three weeks. Isolates of common morphotaxa were lost preferentially over those of rarer taxa. Such losses of isolates may be acceptable if only presence/absence data are collected. If frequency data are vital for community analysis, only fresh material should be utilised. Two surface sterilisation treatments were applied to N. dealbata leaves. An ethanol/sodium hypochlorite treatment was considered unsuitable because it significantly reduced the number of morphotaxa derived from the leaf lamina. A sodium hypochlorite treatment reduced the number of detectable propagules in the wash water without changing the number of morphotaxa derived from leaf particles in comparison with those of the control group.

The fungus first described as Lentinus flabelliformis var. herbarum has been found fruiting on dead Epilobium angustifolium stalks at La Chaise-Dieu (France), the first record since its original diagnosis. The basidiospore characters, trama and hymenium structure indicate the species belongs Lentinellus sect. Omphalodei (Hericiales), but the absence of clamps is a remarkable character, new for this genus. The new combination L. herbarum comb. nov. (syn. Lentinus flabelliformis var. herbarum) is made and a neotype designated. A key to the eight European taxa of sect. Omphalodei is included.

Palaeoclavaria burmitis gen. et sp. nov. (Palaeoclavariaceae fam. nov., Hymenomycetes) is described from a series of fruit bodies and hyphae in Cretaceous amber from Burma (about 100 Myr). This is the first fossil record of the Aphyllophorales and establishes certain basic morphological and ecological characters for the group.