Strains of Trichoderma harzianum possess biocontrol capabilities. As background for identification of strain-specific markers for monitoring strains of interest, the relationship of strains designated as T. harzianum, including representatives of three biological forms Th1, Th2 and Th3, were analysed by Universally Primed PCR, UP-PCR, using UP and random primers. Cross dot blot hybridization of UP-PCR products generated with either of two different UP primers or a random primer showed unequivocal differences among strains. Using this approach, T. harzianum strains were distributed into two different genotypic groups. One of the T. harzianum groups included forms Th1 and Th2 while the other group accounted for the Th3 form. The relatedness of strains of each group was estimated by UPGMA analysis based on markers revealed with three primers. It was found that both genotypic groups are heterogeneous, and Th2 form strains definitely cluster together with those of Th1. Division of the three biological forms of T. harzianum into two groups was also supported by rDNA-ITS1 analysis, where Sau3A digestion of the amplified ITS1 region gave a restriction fragment profile specific for each genotypic group. Two strains with known biocontrol capabilities were found to relate to the genotypic group containing Th1 and Th2 forms and, based on variation within this group, to belong to a homogeneous group of form Th1 strains. The robustness and reliability of UP-PCR fingerprinting were demonstrated by obtaining identical banding profiles using different conditions for PCR in different laboratories.

Fungal research and education has for many years been supported by public service genetic resource centres, whose roles have been to maintain, preserve and supply living cultures to the research community. In the genomic era, genetic resource centres are perhaps more important than ever before. The cultures held, many of which are described and validated by expert biosystematists, are valuable resources for the future. There is a need to supply genomic and proteomic research programmes with fully characterised organisms, as usage of organisms from unreliable sources can prove disastrous, not least in economical terms. However, mycologists often require more than just the organisms, for example, their associated information is vital for bioinformatic applications and some researchers may only require genomic DNA from the organism rather than the organism per se. Genetic resource centres are continually adapting to meet the needs of their users and the wider mycological research community, this associated with OECD international initiatives should ensure they exist to support research for many years to come. This review considers the impact of such initiatives, the current roles of fungal genetic resource centres, the mechanisms used to preserve organisms in a stable manner and the range of resources that are offered for genomic research.

The Lulworthiales consists of four genera: three that were removed from the Halosphaeriales, namely Lulworthia, Lindra, and Kohlmeyeriella; and Spathulospora, reassigned from the Spathulosporales. However, studies have shown that neither Lulworthia nor Lindra are monophyletic genera. This study was therefore undertaken to re-evaluate the genera of the Lulworthiales based on the SSU and LSU rDNA genes. Taxonomic revisions are proposed here for Lulworthia crassa, L. lignoarenaria, L. uniseptata and Lindra marinera: Lulworthia crassa is transferred into the genus Kohlmeyeriella; Lulwoidea gen. nov. is established for L. lignoarenaria; Lulwoana gen. nov. is established for L. uniseptata; and Lindra marinera is reduced to synonymy with L. thalassiae. Taxonomic descriptions are emended for the genus Lulworthia s. str., and for L. grandispora and Lindra thalassiae. A neotype is designated for Lulworthia grandispora.

Anthracnose caused by Colletotrichum is a severe foliar disease in rhododendron plantations in Sweden and Latvia. Isolates of this pathogen were collected and characterised based on morphological and molecular criteria. Out of 37 isolates examined, two with falcate spores were identified as C. dematium. The remainder of the isolates had straight cylindrical conidia, which were too variable to be reliably assigned to C. gloeosporioides (syn. C. azaleae) the reported causal agent of anthracnose of Rhododendron spp. These isolates were compared with the reference strains of C. gloeosporioides, with holo- and paratypes and with reference isolates of C. acutatum. The light and scanning electron microscopy examinations revealed that conidial shape was not a reliable criterion for separation of these two taxa and for the identification of our isolates. The two internal transcribed spacers (ITS1 and ITS2) and the 5·8S rRNA gene were PCR amplified and sequenced from all the rhododendron isolates and the reference strains, including the holo- and paratype materials. In addition, a segment of mitochondrial small subunit ribosomal DNA (mtSSU rDNA) and a fragment of a β-tubulin gene from several representatives of isolates of rhododendron as well as the reference strains were amplified and sequenced. The sizes of the amplified fragments among the isolates studied were heterogeneous only for mtSSU rDNA, which allowed separation of C. acutatum from C. gloeosporioides. Parsimony analysis of the individual and combined nucleotide sequence data sets were concordant and indicated that all isolates originating from rhododendrons belonged to C. acutatum. The grouping of the isolates was further supported by bootstrap analysis. Thus, C. acutatum was identified as the causal agent of anthracnose of Rhododendron in Sweden and Latvia. To our knowledge, this is the first report of C. acutatum and C. dematium on Rhododendron. This study is also the first to determine the genetic status of the holotype of C. acutatum and also one of the paratypes on the basis of ITS sequences in comparison with world-wide living isolates of this taxon.

Penicillium brevicompactum and other isolates with the compact, complex conidiogenous apparatus typical of the species were sequenced in one ribosomal and two protein coding regions. The aligned DNA sequences were analyzed by maximum parsimony and the data from different loci were tested for compatibility using the partition homogeneity test. Analysis of each of the three loci revealed three clades corresponding to P. brevicompactum, P. olsonii and P. biourgeianum. Using the phylogenetic species concept and the genetic isolation of the clades, P. hagemi, P. patrismei, P. stoloniferum, and P. griseobrunneum are all synonyms of P. brevicompactum. P. volgaense is a synonym of Penicillium olsonii, while P. biourgeianum is a distinct species closely related to P. brevicompactum and P. olsonii. Phenotypic distinctions between the species are mostly based on colony characteristics such as colour. P. bialowiezenze, often treated as a synonym of P. brevicompactum, is most closely related to P. polonicum.

Arctic and temperate strains of Hebeloma spp. were grown in axenic culture on glutamic acid, alanine, lysine and NH4+ as sole sources of nitrogen (N), with excess carbon (C) or deficient C (supplied as glucose). Their ability to utilize seed protein as a natural N source was also assessed. All strains tested had the capacity to assimilate amino acids and generally utilized alanine and glutamic acid more readily than NH4+. Some strains were able to utilize amino C when starved of glucose C, and could mineralize amino-N to NH3-N. Arctic strains, in particular, appeared to be pre-adapted to the utilization of seed protein N and glutamic acid N, which is often liberated in high concentrations after soil freezing. The results are discussed in relation to their possible ecological importance.

Anzia electra sp. nov. is described and illustrated from Baltic amber dating back approx. 40 Myr. The diagnostic features of the fossilised species include the small, narrow lobed foliose thallus, two-layered medulla, lack of chondroidal axis, moniliform spongiostratum, and rhizines born singly at the margins of spongiostratum cushions. The fossil species demonstrates that all distinguishing features in the thallus morphology of Anzia sect. Anzia have remained stable for tens of millions of years. Hence the divergence of anzioid lichens must have occurred in the distant past, most probably before the Tertiary. The fossil also provides the first hard evidence that the disjunct-Laurasian distributions of some modern lichens represent relicts of a formerly wider range. The close similarity of the fossil to extant species in East Asia and eastern North America suggests that the present range of Anzia sect. Anzia is highly relict.

A method is described for identifying strains of Metarhizium suitable for use in field experiments. It involves the restriction endonuclease digestion of a PCR product derived from the Pr1 protease gene and the analysis of the fragments by electrophoresis. Using this technique, 40 Metarhizium strains produced 15 different profile types and were clustered into four groups. Correlation between the profile of restriction fragments and geographic origin was observed for certain groups of strains. This PCR strategy allowed the identification of fungal strains, using as samples spores scraped from the surface of single insects killed by the fungus or single whole dead insects with external mycelium. The sequence of the Pr1 PCR product from two strains revealed that Pr1 gene has at least three introns.

The cultivation and competitiveness of Tuber borchii was investigated. Seedlings of Pinus pinea were infected either with T. borchii, Laccaria bicolor (strain S238), Hebeloma sinapizans, T. borchii and L. bicolor, or T. borchii and H. sinapizans. Uninoculated seedlings weakly infected with Sphaerosporella brunnea were used as controls. In 1990 the seedlings were transplanted to a coastal site at Marina di Ravenna, Italy. Examination of seedling roots in 1994 and 1995 showed that L. bicolor, H. sinapizans and S. brunnea ectomycorrhizas were replaced by T. borchii and other ectomycorrhizal fungi. The DNA sequence of the ITS region was analysed using the specific pair of primers, ITS1–ITS4, to type the ectomycorrhizas of Tuber borchii. The ectomycorrhizas were also identified by digestion of the PCR products using different restriction enzymes. T. borchii truffle production began in 1994 and for the next four years an average of 7 kg of truffles ha−1 were harvested. This study describes a simple method for the cultivation of T. borchii.

Phytophthora megakarya is an important pathogen of cocoa in Africa. We used isozyme and RAPD markers to estimate the genetic diversity and structuring among 161 isolates, from the known distribution area of the fungus which corresponds to the cocoa belt in Ghana, Togo, Nigeria, Cameroon, Gabon and Sao Tome. Thirty six and 44 multilocus patterns were identified with isozymes and RAPDs, respectively. Patterns were separated into two highly differentiated genetic groups with both types of markers, one located in Central Africa and the other in West Africa. This distribution coincides with two major biogeographical domains which may reflect an ancient evolution of P. megakarya in this part of Africa. The genotypic diversity was lower in West Africa as compared to Central Africa. Inside Central Africa, isolates from Gabon and Sao Tome were highly differentiated based on RAPDs. Four intermediate marker patterns corresponding to isolates sampled near the border between Nigeria and Cameroon were putatively derived from genetic exchanges between the two major groups. The mating type determination permitted to confirm the high prevalence of A1 over A2. Although clonal multiplication seems to be the rule, indices of other reproduction means have been detected.

Considerable taxonomic confusion exists regarding many species of Cantharellus. Several taxa have been classified in either Cantharellus or Craterellus depending on which morphological characters were emphasised. Recent phylogenetic studies based on sequence analyses suggest that some species now classified in Cantharellus should be in Craterellus. We extracted DNA from dried herbarium specimens and amplified, sequenced and cladistically analysed approximately 650 bp of the 5′ end of the nuclear large subunit ribosomal DNA gene. Our data confirm earlier results and provide molecular evidence for the classification of additional Cantharellus species (Ca. ignicolor and Ca. lutescens) within Craterellus. Furthermore, the data enable us to predict that all Leptocantharellus-like species currently classified within Cantharellus are more accurately classified within Craterellus. The species complexes of Cr. tubaeformis and Cr. cornucopioides were also investigated. These analyses did not support the monophyly of Ca. tubaeformis but the hypothesis that colour varieties of Cr. cornucopioides represent separate species was rejected.

Sowerby described Agaricus cossus in 1799. The fungus possessed a smell, resembling that of a wounded larva of Cossus cossus (Lepidoptera). The species belongs in Hygrophorus, and since more than one white Hygrophorus species has this distinctive smell the epithet cossus has been variously interpreted. The complete internal transcribed spacer (ITS) region of the original type collection made in 1794, preserved in the Royal Botanic Gardens Kew herbarium, was successfully sequenced. Comparison with the ITS sequences from four other white aromatic-acidulous smelling Hygrophorus species, including the type specimen of H. quercetorum, showed that H. cossus is a species associated with Quercus and an older name for H. quercetorum. The differences in basidiome colouration developing with age and host-tree association appear to be the most useful characters to discriminate between the four species with a Cossus cossus smell. A table of morphological and ecological characters is provided.

The tef1 gene coding for translation elongation factor EF1α was identified by a PCR-based approach using degenerate oligonucleotides derived from conserved regions seen in alignments of other EF1αs. The deduced amino acid sequence was compared to other EF1αs. In Southern blot analyses no evidence for a second gene encoding EF1α in Schizophyllum commune was detected. Abundant transcript levels were detected in Northern blot analyses. The open reading frame is interrupted by seven small introns ; however, the positions of these introns are not conserved between the homobasidiomycete Schizophyllum commune and the heterobasidiomycete Puccinia graminis genes.

Eutypa lata, the causal agent of Eutypa dieback of grapevines, is difficult to identify on the basis of colony morphology and is often out-competed by other fungi when isolated from wood. To facilitate diagnosis of the pathogen, we designed SCAR primers capable of amplifying DNA of E. lata and constructed a genomic DNA library from which DNA sequences specific to E. lata were identified and sequenced. SCAR primers were used to identify E. lata directly from culture without the requirement for DNA extraction or prolonged incubation periods and could also detect the pathogen in DNA isolated from grapevine wood. RFLP probes were used in slot-blot assays to detect the pathogen in DNA isolated from 1 yr old cane as well as from mature grapevine trunks. The markers developed in this study have the potential to be used as a research tool to gather information on the epidemiology of the disease and to assess the efficacy of potential control agents against E. lata.

The phylogenetic relationships of the stem rot pathogens Sclerotium rolfsii and S. delphinii were examined, based on their rDNA ITS sequences. The ITS regions were cloned and sequenced to identify three distinct ITS types: r-1, r-2, and r-3. Two different ITS types exist within S. rolfsii and S. delphinii strains. Japanese strains and one strain of S. rolfsii from the USA contain types r-1 and r-2, whereas another strain from the USA and one from Chile have only one ITS type, r-2. S. delphinii strains have types r-1 and r-3. We discuss the implications of the common presence of ITS type r-1 for the taxonomy and evolution of this species complex.

All hypocrealean species of the genus Geosmithia are anamorphic fungi with connections to bark beetles. G. fassatiae, G. langdonii and G. obscura are described as new sympatric species associated with Scolytus carpini, S. intricatus and S. rugulosus in Central Europe. The species represent a complex of three sister taxa with affinities to G. flava that may be distinguished by differences in morphology, unique RAPD patterns and by sequences of ITS region rDNA. Intraspecific variability and habitat specificity of new species is described and discussed. The high morphological, genetic and ecological uniformity suggest that these Geosmithia spp. are recently derived. A key to all accepted hypocrealean species of the genus is provided.

Mating type and RG57 fingerprints were determined for 141 isolates of Phytophthora infestans from 63 fields in all the important potato growing areas in Norway and Finland. Seventy-six multilocus genotypes were identified, of which 53 were detected only once. Norwegian and Finnish isolates were similar in terms of genotypic diversity and the genetic distances between the genotypes. The large number of genotypes and the high genetic distances between the genotypes indicate that sexual reproduction is contributing significantly to the genetic variation of P. infestans in Norway and Finland.

The mycoparasite Trichoderma harzianum, a biocontrol agent, forms coils and other mycoparasitic structures upon contact with host fungi. A biomimetic system consisting of nylon fibres was used to test the involvement of signal transduction pathways in the induction of coils. Two activators of G protein-mediated signal transduction induced coiling of hyphae around nylon fibres. The peptide toxin mastoparan increased coiling more than two-fold in comparison with controls. The activator fluoroaluminate (A1F4−) had a similar effect, whereas aluminium ions alone were ineffective. cAMP increased coiling about three-fold. Although the two G protein activators, mastoparan and fluoroaluminate, have very different modes of action, they share the Gα subunit as a target. We also found that two T. harzianum isolates differ in their extent of coiling around nylon fibres in the absence of effectors. The results reported here demonstrate that the biomimetic system can be used to study the biochemistry of coil induction, and will be a valuable assay to aid in the genetic manipulation of this pathway. It is proposed that a signal for mycoparasitic behaviour from the host cell surface is transduced by heterotrimeric G protein(s) and mediated by cAMP.

The saprotrophic wood-degrading abilities of strains from the intersterility groups S and P of the necrotrophic root and white rot fungus Heterobasidium annosum were tested using a conventional soil jar method and small wood blocks of Pinus sylvestris, Picea abies and Betula verrucosa. Weight loss in dry matter of wood blocks was compared with the ability of the same strains to secrete the phenol oxidase laccase under liquid and solid-state culture conditions. Results showed the much greater ability of H. annosum P strains to degrade wood, with weight losses comparable to those reported for other white rot fungi cultivated under similar conditions. In contrast, the S isolates except Br228 and Fr154 degraded the wood blocks poorly with a maximum weight loss of ca 12% recorded after 5 mo incubation. Light and scanning electron microscopy showed hyphal colonization and decay to be typical for white rot, with both simultaneous attack by cell wall thinning and preferential lignin degradation recorded for P strains. Results for wood block degradation correlated well with the ability of the intersterility groups to produce laccase in liquid culture and solid-state culture conditions, with P strains producing ca 5–6 times more laccase than S strains. Results indicate that great differences exist between H. annosum intersterility P and S groups ability to cause wood decay, but that P strains have a significantly greater competitive saprotrophic wood-degrading ability than previously realized.

Filamentous hyphal growth is inherently suited to kinetic analysis, and in many respects the fungal mycelium can be viewed as a very mechanical biological system, which lends itself to mathematical modelling. The mathematics of hyphal tip extension growth are well-established. However, even though a hyphal growth equation can be written with confidence, and we have a good understanding of the effects of tropisms on growth, it is not easy to form a mental picture of the behaviour of large populations of hyphal tips. What is required, and what we believe we have produced, is a mathematical model that is sufficiently sophisticated to produce a realistic visualization of fungal hyphal growth. This provides us with a cyberfungus that can be used for experimentation on the theoretical rules that might govern hyphal patterning, hyphal interactions, and tissue formation and organ development by actually visualizing the virtual hyphal growth patterns that result from different regulatory scenarios. From a series of model experiments the most significant observation is that complex fungal fruit body shapes can be simulated by applying the same regulatory functions to all of the growth points active in a structure at any specific time. No global control of fruit body geometry is necessary. No localized regulation is necessary. The shape of the fruit body emerges from the concerted response of the entire population of hyphal tips, in the same way, to the same signals.