The mycorrhizal fungi in the roots of achlorophyllous Sciaphila japonica and S. tosaensis (Triuridaceae) were identified by molecular methods. The habitats of S. japonica were in a tree plantation of Japanese cypress, Chamaecyparis obtusa, and bamboo forests, and those of S. tosaensis were in a camellia forest and a bamboo forest. In the root cortical cells of both plants, aseptate hyphal coils were observed, which suggested the Paris-type arbuscular mycorrhiza (AM). A phylogenetic analysis based on a partial sequence of an AM fungal nuclear small subunit ribosomal RNA gene showed that the fungal DNA sequences of S. japonica were separated into three closely related clades. Those of S. tosaensis were separated into two clades, which were also closely related to each other. The AM fungi of S. japonica and S. tosaensis were completely separated in the phylogenetic tree even among those found in the same habitat, which suggests the high specificities in the plant-fungal partnerships. All the detected AM fungi in these plants belonged to Glomus-group A. Even though the habitats are in quite common environments, both plant species are known as endangered species in Japan. Such a definite specificity in AM symbioses seems to restrict the distribution of the myco-heterotrophic plants.

In this study, sequence-related amplification polymorphism (SRAP) and inter-simple sequence repeat (ISSR) were analyzed for accessing the genetic diversity of 37 Monascus isolates and 14 control strains. According to the dendrogram produced by SRAP data, all the tested strains were grouped into four clusters at a 78% similarity level. Comparatively, 51 tested strains were divided into four major groups at a similarity level of 74% based on the dendrogram generated via ISSR marker analysis. Based on the two sets of dendrograms, Monascus aurantiacus, M. purpureus, M. serorubescens, M. anka, and M. ruber were clustered in the same clade; M. albidus, M. fuliginosus, and M. barkeri were clustered with M. pilosus in a second clade; and M. lunisporas and M. argentinensis occurred together in a third cluster distinct from the other Monascus species. The cluster result produced by SRAP data shared great similarity with that by ISSR data with minor differences in the subgroups, which is basically in agreement with morphological observations. In general, SRAP and ISSR are more simple, rapid, and efficient, which may provide alternative molecular approaches to studying genetic diversity, classification, and identification of Monascus strains.

The protease activities in six edible mushrooms were surveyed using synthetic fluorogenic substrates that have different specificities for each protease group. The activity was determined by measuring the fluorogenic intensity of the 7-amino-4-methylcoumarin (AMC) liberated by an enzyme. Various types of activities were found in all mushrooms, and their activities depended largely on the mushroom species, but also on the pH and localization. Flammulina velutipes and Pleurotus eryngii had the widest and highest proteolytic activities among the six mushrooms examined. The proteasome-like protease activities were generally much higher than those of other proteases. High caspase activities, which occur during apoptosis in cells, were detected in two mushrooms, F. velutipes and Hypsizigus marmoreus. The pH optima of the proteolytic activities were largely divided into two groups, acidic pH 5–6 for caspases and neutral to alkaline (pH 6.5–11) for the others. In F. velutipes, higher proteolytic activity was observed in the basement of the stem than in the cap and stem. Purification and characterization of protease were also carried out to identify a protease from Grifola frondosa using t-butyloxycarbonyl-Leu-Arg-Arg-4-methylcoumaryl- 7-amide (Boc-LRR-MCA) as the substrate.

To study thraustochytrids from temperate and cold environments of Southern Argentina, the standard cultivation methodologies have been modified because many of the microorganisms detected by microscopic examination in both the original samples and the colonized baits failed to be successfully isolated in standard culture media. As a result, 35 strains, most of them having a very low growth rate, were isolated. Alternative procedures are proposed according to the nature of the sample, the characteristics of the thraustochytrid to be isolated, and the presence of contaminating microorganisms. Modifications proposed include the use of a newly formulated culture medium (Mar Chiquita, containing glucose, gelatine hydrolysate, peptone, and corn steep liquor as main carbon and nitrogen sources). In addition, the effects of the nutrient composition and agar concentration of culture media on the relative growth rates of the isolates were studied in an attempt to determine the most suitable conditions for the cultivation of new strains of thraustochytrids. The goal of this study is to develop a standard methodology, allowing us to grow baitable “elusive” thraustochytrid strains, and that could be applied to improve the isolation and the study of the undocumented biodiversity of this group of microorganisms from different environments.

Cercospora yakushimensis is proposed as a new combination, Pseudocercospora yakushimensis comb. nov.,and the morphology of the species is characterized from herbarium specimens, including the type. Six additional species belonging to the genus Cercospora and its allied genera are newly added to the Japanese mycoflora, namely,Cercospora dioscoreae-pyrifoliae on Dioscorea tokoro,Pseudocercospora catalpigena on Catalpa ovata, Pseudocercospora coriariae on Coriaria japonica, Pseudocercos pora lythri on two species of Lythrum, Pseudocercospora pteroceltidis on Celtis boninensis, and Passalora puncta on Foeniculum vulgare.

Paecilomyces tenuipes is a popular medicinal mushroom, and has received extensive attention for medical application because of its various physiological activities. However, there is limited information about the anticancer and immunomodulatory activities of Paecilomyces tenuipes. This study attempted to evaluate the effect of an extract from the cultured fruiting bodies of Paecilomyces tenuipes (PTE) on the expression of the interleukin- 18 (IL-18) gene in rat pheochromocytoma (PC) 12 cells and rat brain. Related mRNA levels were determined by reverse transcription-polymerase chain reaction (RT-PCR). Protein levels were measured by Western blot and immunohistochemistry. Our results demonstrated that PTE induced IL-18 gene expression both in vitro and in vivo. Treatment of PC12 cells and rat brain cells with 10 μg/ml and 20 mg/kg PTE, respectively, yielded significant increases of IL-18 levels. Significantly, IL-18-immunoreactive neurons were detected in the Purkinje cells of the cerebellum. IL18-immunohistochemical staining was markedly enhanced in animals treated with PTE compared to findings in the untreated controls. These results suggest that PTE could be a potential candidate as an immune activator or anticancer drug.

There were no published data about the occurrence of Libertella species in Slovakia. Three Libertella species, L. faginea on Fagus sylvatica, L. betulina on Betula pendula, and L. quercina on Castanea sativa, are described here. Morphological observations with descriptions of other Libertella species growing on Fagaceae were compared, and possible conspicuity with any of the known species is discussed.

Thirteen strains were isolated from different habitats, belonging to two genera, namely Conidiobolus and Basidiobolus, related to saprophytic Entomophthorales. Chitin flake colonization and agar-well diffusion tests were used to screen potential extracellular chitinase-producing strains in plate assays. Preliminary screening resulted in five chitinase producers that were further studied quantitatively. Results indicated that studied isolates of this group produced chitinase at different levels in chitin-containing as well as non-chitin-containing medium. Conidiobolus coronatus was found to be the most significant chitinase producer, giving 0.261 U/ml using colloidal chitin as a carbon source, among the isolates under study. This communication also reports the chitinolytic activity of Basidiobolus haptoporus, the effect of environmental and nutritional parameters on chitinase production, and utilization of fungal biomass as a carbon source, which hitherto had not been elaborated from this genus.

Reverse complementary DNA sequences—sequences that are inadvertently cast backward and in which all purines and pyrimidines are transposed—are not uncommon in sequence databases, where they may introduce noise into sequence-based research. We show that about 1% of the public fungal ITS sequences, the most commonly sequenced genetic marker in mycology, are reverse complementary, and we introduce an open source software solution to automate their detection and reorientation. The MacOSX/Linux/UNIX software operates on public or private datasets of any size, although some 50 base pairs of the 5.8 S gene of the ITS region are needed for the analysis.

Hydrophobins secreted by filamentous fungi self-assemble into an amphipathic film at hydrophilic/ hydrophobic interfaces. This unique property suggests that the hydrophobins have a high potential for industrial applications. However, the assemblages of class I hydrophobins are highly insoluble, making such commercial applications difficult. To enhance the solubility of class I hydrophobins, we have attempted to express class I hydrophobin PNH1 from Pholiota nameko fused with glutathione S-transferase (GST) in Escherichia coli. The GST–PNH1 was effectively isolated from the soluble fraction of transformed E. coli, and subsequent analysis revealed that the purified GST–PNH1 had almost the same emulsifying activity as PNH1.