The entomopathogenic fungus Metarhizium anisopliae is generally considered to be mesophylic, although some isolates have the ability to grow at 8 °C. In two cold-active isolates (DAT-1 from Tasmania and CLB2-1vi from Ontario, Canada) and one non-cold-active isolate (Ma 2575 from South Carolina), we (1) analyzed cold-induced intracellular proteins, using two-dimensional (2D) gel electrophoresis, and (2) analyzed cold-active transcripts, using a polymerase chain reaction (PCR) based subtractive hybridization technique. Protein differences, as observed by 2D gel electrophoresis, were observed in cold active isolates grown at 8 ° when compared with 25 °, but such differences were not observed in the non-cold active isolate. Fungi were also subjected to various stresses (45 °, pH, and salinity) to determine how proteins induced under these conditions compared to proteins induced under low temperature growth. Proteins induced were either specific to cold-active growth or were general stress proteins and differences were also observed in 2D patterns between the two cold-active isolates. Transcripts upregulated during growth at 8 ° and isolated by a PCR based subtractive hybridization technique also differed between the two cold-active isolates. Analysis of the transcripts showed several novel sequences but also included transcripts with similarities to actin, NADPH quinone oxidoreductase, a thiamine biosynthesis protein and a yeast-like membrane protein. The potential role of these proteins in cold-active growth is discussed.

Haptoglossa dickii is a biflagellate zoosporic parasite of bactivorous nematodes. The walled encysted zoospores germinate more or less synchronously to produce specialised infection cells known as gun cells. This paper documents the changes in the main cytoplasmic organelles, namely the nucleus, Golgi body, mitochondria, lipid droplets and dense-body vesicles, from the onset of encystment until the formation of the mature gun cell. The septum-delimited gun cells show a marked polarity in the distribution of their organelles from their inception, which seems in part determined by the temporal order by which the main organelles migrate out of the cyst and into the gun cell initial. In this species the majority of dense-body vesicles and mitochondria precede the nucleus and lipid droplets. The nucleus remains more or less central throughout the remaining course of gun cell differentiation. The single nucleus associated Golgi body changes its orientation to track (possibly even determining) the spatial position of the ingrowing injection tube. The mitochondria fuse to form one or more complex reticulate structures which are aligned close to the injection tube throughout its formation and which in the mature cells form a fine network entwined around the fully differentiated tube. The vesicles containing electron-dense inclusion granules (dense-body vesicles) are initially concentrated towards the gun cell anterior, but later migrate to the posterior end of the cell and fuse to give rise to the basal vacuole. Some of the likely functional aspects of these cytoplasmic changes are discussed.

The Cainiaceae is revived to accommodate Atrotorquata, Arecophila, Cainia, Ceriophora and Reticulosphaeria, all of which are described and illustrated. Ommatomyces is also included in the family and discussed. The essential characteristics of the family are ascomata which are usually immersed under a clypeus, unitunicate asci with a complex, J+, apical apparatus comprising a series of rings, and brown ascospores which may have longitudinal germ slits, striations, reticulate ornamentations and/or germ pores.

Urediospore-derived infection structures of Phakopsora apoda, a rust fungus on kikuyu grass (Pennisetum clandestinum), penetrate the host leaf directly through the cuticle and not through the stomata. A urediospore germinates on the leaf to form a short germ tube which is delimited from a terminal appressorium by a septum. Most appressoria form at the junctions between epidermal cells. Appressoria are often sessile to the urediospores. From the base of the appressorium, a penetration peg develops, which penetrates through the host cuticle and epidermal cell wall. Penetration of the epidermal cell wall occurs approx. 6 h post inoculation. Once inside the epidermal cell, the penetration peg expands to form a penetration hypha, which traverses the epidermal cell and emerges into the intercellular space of the mesophyll tissue. A septum is formed in the intercellular portion of the penetration hypha, delimiting a primary hypha which extends further into the mesophyll, before branching to form two secondary hyphae. This penetration process appears to be very similar to that of Phakopsora pachyrhizi on soybean.

Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis of the first intergenic region (IGR1) of nuclear ribosomal DNA was used to clarify the relationship between IGR1 variations and six Japanese biological species of Armillaria: A. gallica, A. nabsnona, A. ostoyae, A. cepistipes, A. mellea and Nagasawa's E (Nag. E: taxonomically unknown species). The procedure was then used to identify Armillaria species associated with Wynnea species (W. americana and W. gigantea) and Entoloma abortivum. By combining the RFLP patterns obtained using three endonucleases, HaeIII, HinfI and MspI, the IGR1s from 18 isolates of six Armillaria species were assigned to nine different RFLP phenotypes and the six species were distinguished from each other. Each of the RFLP phenotypes from the Armillaria isolates associated with Wynnea species or E. abortivum matched a corresponding phenotypes observed among the six Armillaria species. Based on this, all four isolates from W. gigantea were identified as A. mellea, two from W. americana as A. cepistipes, and all three from E. abortivum as Nag. E. These results provide new information on the biological species of Armillaria associated with Wynnea and E. abortivum.

Beauveria bassiana is an entomopathogenic fungus with a growing potential for pest control in different agro-ecosystems worldwide. Such potential brings the necessity of developing a strain specific typing system. In a previous study, we reported the identification of molecular variants in mitochondrial DNA (mtDNA) polymorphism in 15 North American isolates. Results indicated a highly conserved mitochondrial genome showing only two mitochondrial genotypes (mitotypes). In this study we used whole genomic DNA from 18 isolates of B. bassiana, two unidentified Beauveria spp., and one each of B. amorpha, B. cylindrospora and B. nivea from more diverse origins. By doing single- and double-restriction enzyme digestion of total genomic DNA with EcoRI, and HindIII and then probing with BbmtE2, the predominance of mitotypes A and B was observed again, along with three newly described mitotypes (C to E). Additionally, by using whole B. bassiana mtDNA digested with HpaII as probe, we further demonstrate up to nine different mitotypes within B. bassiana. With either of the two probes, distinguished between members of the genus Beauveria and from Paecilomyces farinosus and Metarhizium anisopliae. Phylogenetic analysis could not however distinguish B. amorpha and B. nivea isolates from B. bassiana, suggesting a close genetic relation between the three species of the genus. Altogether, these results show high variability in mitochondrial genome, which can be useful as a reliable tool for the biopesticide industry for both species and isolate specific identification.

The identification of Trichoderma strains is important for their application as biocontrol agents. We used a two-dimensional analysis, in which extracellular proteins of Trichoderma harzianum strains T-35, Y and TM were first separated according to their isoelectric point and then according to their molecular mass. Chitinase activities were detected in situ after the second separation. Each of the three strains exhibited a unique pattern of 3–5 different chitinases (1 or 2 N-acetyl-β-glucosaminidases, and 2 or 4 endochitinases). These unique profiles can be used to differentiate among strains within this species. The suggested procedure makes use of a property of T. harzianum that is directly involved in the mycoparasitic interaction, making it suitable for specific biocontrol applications.

To test the validity of R. tauricus as a separate species, its carbon source utilization, isoenzyme patterns and PCR-coupled RFLP of the ITS of its rDNA were analysed. Whilst R. tauricus differed considerably from R. pusillus and R. miehei in morphological traits as well as its ability to row on single carbon sources, the results obtained from all other experimental approaches suggest extensive similarity between R. tauricus and R. pusillus. These findings lead us to believe that R. tauricus is a mutant heterothallic strain of R. pusillus.

Crossing and selfing experiments were carried out with six field collections and five isolates belonging to three formae speciales, larici-epitea typica (LET), larici-daphnoides (LD), and larici-retusae (LR), of willow rust, Melampsora epitea. European larch (Larix decidua) was inoculated with basidiospores produced on overwintered telial leaves and the resulting spermagonial lesions on larch needles subsequently paired in vitro. In two crosses between LR and LD involving a total of 439 lesion pairs, only two cultures obtained were identified as hybrids. These two cultures were non-pathogenic to the maternal host Salix burjatica cv. Korso and weakly pathogenic to the paternal host S. daphnoides cv. Meikle. No identifiable hybrids were obtained from 56 lesion pairs between LET and LD. In a cross between LET as receptor and LD as donor, one-third of the lesions formed aecia. In the reciprocal combination, however, less than 1% developed aecia. All F1 cultures between LET and LR were weakly pathogenic to the parental hosts S. viminalis cv. Mullatin and S. burjatica cv. Korso. When three of the F1 cultures were used to produce telia, only one developed mature teliospores. Subsequent selfing experiments showed that this culture was predominantly self-sterile, with only 1% of lesions producing aecia. In selfing and crossing two pathotypes within the same f. spp., 20–40% of needles produced aecia and the rate of aecial formation was similar in both directions. The results obtained suggest that M. epitea is heterothallic and the sexual compatibility is controlled by a pair of alleles at a locus. It is concluded that the three f. spp. are genetically different populations and ecologically fit new pathotypes cannot arise easily as a result of hybridization between them. Within a f. sp., however, many pathotypes exist or will occur due to exchange of genes for virulence during the sexual life-cycle.

Pseudocercospora mori produced three kinds of hyphae – primary infection hyphae, internal hyphae and secondary infection hyphae. The primary infection hyphae were developed from conidia and they penetrated the leaf through stomata directly or by lateral penetration branches – the stomatopodia. Inside the stomatal chambers the stomatopodia swelled to become vesicle-like structures. The internal hyphae produced compact stromata in the sub-stomatal chambers. Each stromata carried 1–5 conidia at a time on short unbranched conidiophores. The secondary infection hyphae were developed from internal hyphae directly or from stromata. The secondary infection hyphae emerged through stomata, branched profusely on the leaf surface and produced conidia singly on short conidiophores. The secondary infection hyphae re-entered the leaf directly or by stomatopodia. Conidia were developed also from stomatopodia and from their substomatal vesicles. The conidia developed from substomatal stromata, and those produced from the secondary infection hyphae on leaf surface were similar in their morphology and size of conidiophores. The conidia were obclavate to cylindrical, 3–17 celled and measured 4–35×1.5–2 μm. Germ tubes were often found to emerge from the conidial cell nearer to stomata. The primary infection hyphae and secondary infection hyphae produced stomatopodia above stomata.

Paraphaeosphaeria barrii, a new species of the genus found on Agavaceae, is described from leaves of Yucca schidigera collected in Baja California, Mexico. The Coniothyrium anamorph is briefly described from cultured ascospores. This species was found to be most closely related to P. conglomerata, P. filamentosa, and P. quadriseptata all of which are from Agavaceae, produce 3 or 4 septate ascospores and pigmented, non-septate conidia from holoblastic, percurrently proliferating conidiogenous cells.

Heterothallism was confirmed in Mortierella umbellata from Japan. Anisogamic zygospore formation is described and illustrated with time-lapse light micrographs. The anisogamic behaviour of the species is discussed with reference to previous studies on anisogamic zygospore and azygospore formation in Zygomycetes.

Myconymphaea yatsukahoi gen. sp. nov. is described in the Kickxellaceae (Kickxellales, Zygomycetes). The fungus is characterised by unicellular sporocladia formed on apical vesicles of sporangiophores, conspicuously long sporangiospores and peculiar septal plugs with prominent protuberances. In addition, a key to all known genera of the Kickxellales is provided.

The electrophoretic karyotype of the Chinese straw mushroom, Volvariella volvacea, was determined. The haploid strain V34 of V. volvacea has 15 chromosomes ranging in size from 1.4 to 5.1 Mb. No chromosomal polymorphism in terms of size and number was seen in either of two growth stages: vegetative mycelia or fruit-body gill tissues. DNA fingerprints were prepared by the arbitrarily- primed polymerase chain reaction. Those obtained from different stages during fruit-body morphogenesis were identical. However, variation in DNA fingerprints was evident in protoplast regenerants derived from the same vegetative mycelium. Thus the haploid mycelium of strain V34 is heterokaryotic but the bulk genotype is stable during fruit-body development. F1 and F2 progenies germinated from the haploid, uninucleate basidiospores from self-fertilized fruit bodies also regularly segregate a range of mycelial morphological variants as well as phenotypic variation revealed by DNA fingerprints. We overview the known mechanisms to generate genetic variation and propose a novel mechanism that could account for the 1:1 segregation ratio of self-fertile to self-sterile progeny regularly obtained from selfed Volvariella volvacea fruit bodies.

A new species of Penicillium, Subgenus Biverticillium, was isolated from the root of a wheat seedling during a survey of the ability of rhizosphere fungi to solubilize phosphate. The species, described here as Penicillium radicum, has affinities with P. variabile, P. allahabadense and Talaromyces wortmannii, but can be distinguished by differences in morphology, secondary metabolite profiles and unique DNA banding patterns obtained using RAPD PCR.

The following foliicolous fungi on Tabebuia species are described and illustrated: Anhelia tabebuiae sp. nov. and Dictyonella tabebuiae sp. nov. (ascomycetes), Fumagospora tabebuiae sp. nov., Polychaeton tabebuiae sp. nov. and Septoria tabebuiae-impetiginosae sp. nov. (coelomycetes), Cercospora tabebuiae-impetiginosae sp. nov. and Pseudocercospora tabebuiae-roseo-albae sp. nov. (hyphomycetes). Uncinula peruviana was found and described on T. impetiginosa as a first record for Distrito Federal, Brazil.

The ITS region of ribosomal DNA was sequenced for 30 isolates or sporocarps (including five type specimens) representing ten European taxa of Antrodiella. Phylogeny was inferred from the sequences by parsimony with Steccherinum ochraceum as an outgroup. The phylogram is discussed in relation to morphological and ecological characteristics of the species. On the basis of both morphological and molecular data Antrodiella pallasii sp. nov. is described. The study did not support the separation of A. beschidica and A. farinacea from A. semisupina. A. faginea is reported from Fennoscandia.

Haloaleurodiscus gen. nov. (Homobasidiomycetes) is described from Japanese mangrove forests with one species H. mangrovei sp. nov. The genus is morphologically characterized by having resupinate basidioma, nodose-septate hyphae, sulphoaldehyde-positive gloeocystidia, dendrohyphidia and amyloid basidiospores with minute warts. These morphological features are similar to those of Aleurodiscuss. lat., but H. mangrovei differs from the Aleurodiscus primarily in occurring in white pocket-rot and is not closely related based on ribosomal DNA sequence analyses. Molecular data suggest that this species is phylogenetically placed in the root of the ‘Peniophorales’ clade. In addition, ecological and physiological features of the species are provided.

Endophyllum osteospermi is an autoecious, endocyclic rust fungus, which has only been recorded on Chrysanthemoides monilifera ssp. monilifera (Asteraceae, Calendulae), a perennial woody shrub. Both organisms are indigenous to South Africa. Because E. osteospermi is being considered for release in Australia as a biocontrol agent against C. monilifera ssp. monilifera, it was necessary to determine its host range and natural distribution in South Africa. To address this, natural stands of Chrysanthemoides species, as well as other South African asteraceous plants, were monitored for E. osteospermi between 1992 and 2003. A morphological and molecular comparison of specimens referable to Endophyllum was undertaken. Based on these results, E. osteospermi was recorded on C. monilifera sspp. monilifera, pisifera, rotundata, canescens, and subcanescens, C. incana, and an undescribed taxon. E. osteospermi was also recorded on Osteospermum ciliatum, O. polygaloides, and O. potbergense. Furthermore, a closely related but previously undescribed species, E. dimorphothecae sp. nov. is described on Dimorphotheca cuneata. Aecidium elytropappi is transferred to Endophyllum as E. elytropappi comb. nov., being recorded on Elytropappus rhinocerostis and Stoebe plumosa. This study shows that in South Africa E. osteospermi is restricted to a small group of related plant species in the Calenduleae. This rust is therefore considered suitable as a candidate agent for the biocontrol of C. monilifera ssp. monilifera, and pending the results of host specificity testing, would most likely be safe to introduce into Australia.

The new species Umbelopsis dimorpha sp. nov. was isolated from a soil sample in the Red Hills area of Mt Richmond Forest Park, in the northern part of the South Island of New Zealand. It has two kinds of pale pinkish sporangia: (1) single-spored, indistinguishable from those of U. versiformis; and (2) multi-spored, similar to those of U. vinacea. ITS sequences place the species in the immediate vicinity of the former species.