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The fungal mycelium represents the essence of the fungal lifestyle, and understanding how a mycelium is constructed is of fundamental importance in fungal biology and ecology. Previous studies have examined initial developmental patterns or focused on a few strains, often mutants of model species, and frequently grown under non-harmonized growth conditions; these factors currently collectively hamper systematic insights into rules of mycelium architecture. To address this, we here use a broader suite of fungi (31 species including members of the Ascomycota, Basidiomycota and Mucoromycota), all isolated from the same soil, and tested for ten architectural traits under standardized laboratory conditions. We find great variability in traits among the saprobic fungal species, and detect several clear tradeoffs in mycelial architecture, for example between internodal length and hyphal diameter. Within the constraints so identified, we document otherwise great versatility in mycelium architecture in this set of fungi, and there was no evidence of trait 'syndromes' as might be expected. Our results point to an important dimension of fungal properties with likely consequences for coexistence within local communities, as well as for functional complementarity (e.g. decomposition, soil aggregation).

Planting resistant cultivars is the most economical and effective measure to control wheat stripe rust caused by Puccinia striiformis f. sp. tritici (Pst), but the cultivars often lose their resistance due to the emergence of new physiological races. The UV-B-irradiated urediospores of the Pst physiological race CYR32 in China were inoculated on wheat cultivar Guinong 22 for screening virulence-mutant strains. CYR32 and mutant strains (CYR32-5 and CYR32-61) before and after UV-B radiation were used to conduct urediospore morphological and histopathological observations and an investigation of epidemiological components. The results showed that UV-B radiation affected the urediospore morphology of each strain. UV-B radiation inhibited urediospore invasion and hyphal elongation, which mainly manifested as decreases in germination rate, quantities of hyphal branches, haustorial mother cells and haustoria and hyphal length. After wheat cultivar Mingxian 169 was inoculated with the UV-B-irradiated urediospores, the incubation period was prolonged, and the infection efficiency, lesion expansion rate, total sporulation quantity and area under the disease progress curve were reduced. The results demonstrated that CYR32-5 and CYR32-61 may have more tolerance to UV-B radiation than CYR32. The results are significant for understanding mechanisms of Pst virulence variations and implementing sustainable management of wheat stripe rust., (© 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Sclerotium rolfsii is one of the most destructive fungal plant pathogens that can infect over 500 plants and can adapt to diverse environmental conditions. The present research work was carried out to evaluate the impact of both hexa- and trivalent chromium (Cr) on growth, morphology, enzymatic characteristics, and metal accumulation in S. rolfsii under laboratory conditions. Experiments were performed in both malt extract broth and agar growth medium amended with six different concentrations (10, 20, 40, 60, 80, and 100 ppm) of each Cr(III) and Cr(VI) ions inoculated with fungus and incubated for 6-7 days at 25 ± 3 °C. In broth medium, the total protein content was declined and activities of antioxidant enzymes were increased with an increase in metal concentrations. Lower concentrations (10 ppm) of the metal ions stimulated the growth of fungus and higher concentrations (60-100) inhibited it. The Fourier transform infrared spectroscopy (FTIR) assessment showed hydroxyl, carboxyl, and amine groups as major metal binding sites. In agar medium, tolerance index was decreased up to 0.56 at 10-80 ppm of Cr(III) and up to 0.62 at 10-60 ppm of Cr(VI). Considerable modifications were observed in hyphal and sclerotial morphology with an increase in concentration of metal ions. The current study concluded that interference of Cr with growth and physiological process of S. rolfsii could affect its infection level on its host plant. This study provides important information regarding cultivation of susceptible plant varieties in Cr-polluted soil as evidenced by pathogen growth up to 50 ppm of Cr(III) and Cr(VI) ions.

Fomitiporella accommodates polypores producing annual to perennial basidiocarps with an indistinct subiculum (very thin to almost lacking), mostly a dimitic hyphal structure, lacking any kind of setae, with brownish, thick-walled basidiospores, and causing a white rot. Previously, only a few samples of Fomitiporella were studied on the basis of morphological and nuc 28S rDNA (28S)-based phylogenetic analyses. In this study, we made a comprehensive study on Fomitiporella on the basis of collections from Central America, USA, Europe, and China. The phylogenetic analysis, including 28 nuc 28S rDNA and 29 nuc rDNA ITS1-5.8S-ITS2 (internal transcribed spacer [ITS]) sequences newly generated, discovered 14 new lineages. Combined with morphological evidence, 4 new lineages are described and illustrated as new species, viz., Fomitiporella americana, F. micropora, F. sinica, and F. subinermis; 10 other new lineages, each with a single collection, are still treated as unidentified taxa; three new combinations, viz., Fomitiporella tenuissima, F. chinensis, and F. resupinata, are proposed. In addition, F. inermis is redescribed. A key to the 12 known species of Fomitiporella is provided.

Three new species of Phylloporia from tropical China are described in this study: P. manglietiae, P. pendula, and P. pseudopectinata. Phylloporia manglietiae is characterized by triquetrous to ungulate basidiomata, with 6-8 pores/mm, a monomitic to dimitic hyphal system, and broadly ellipsoidal basidiospores, 3-3.5 × 2-2.5 μm. Phylloporia pendula has small, imbricate, and pendent basidiomata, with 7-9 pores/mm, a dimitic hyphal system, and broadly ellipsoidal basidiospores, 3.5-4 × 2.5-3 μm. Phylloporia pseudopectinata differs from other species of Phylloporia by its applanate basidiomata, with 8-9 pores/mm, a dimitic hyphal system, and subglobose basidiospores, 3-3.5 × 2-3 μm. Phylogenetic analyses, based on sequences from the D1-D2 domains of the 28S gene and the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2 = ITS) of the nuc rDNA, support the classification of the three new species in Phylloporia.

Two new species of Fomitiporia growing on Hippophae trees, F. norbulingka and F. subhippophaëicola, are described from southwest China based on morphological characters and phylogenetic analysis. Fomitiporia norbulingka is characterized by pileate basidiomata, mostly angular pores (6-9 per mm), slightly thick-walled generative hyphae, subglobose to globose basidiospores (6.5-7 × 5.5-7 μm), and absence of cystidioles. Fomitiporia subhippophaëicola is diagnostic by effused-reflexed to pileate basidiomata, angular pores (8-10 per mm), thick-walled generative hyphae, subglobose to obovoid basidiospores (6-8 × 5.5-7 μm), and presence of ventricose to fusoid cystidioles. Phylogenetic analysis inferred from combined sequences including the nuc rDNA ITS1-5.8-ITS2 region, 28S rDNA D1-D2 domains, partial sequences of translation elongation factor 1-α, and RNA polymerase II second largest subunits genes indicated that F. norbulingka and F. subhippophaëicola represent two new lineages which group together with F. hippophaëicola., (© 2016 by The Mycological Society of America.)

Phellinidium, including 13 accepted polypore species mostly with resupinate basidiocarps, is one of the most aggressive forest pathogenic genera. This genus is characterized by the combination of a monomitic hyphal structure, abundant hyphoid setae in the context and trama, and hyaline and thin-walled basidiospores. To explore the relationships among the species of Phellinidium, especially those between forest pathogens and saprophytes, we examined 29 specimens representing all 13 previously known species from Asia, Europe and America from morphological and phylogenetic perspectives. A new genus, Coniferiporia, was found to segregate from Phellinidium for three aggressive forest pathogens, and three new combinations, viz. Coniferiporia qilianensis (the generic type), Coniferiporia weirii and Coniferiporia sulphurascens, were proposed. Phellinidium cryptocystidiatum was treated as a synonym of C. sulphurascens. The circumscription of Phellinidium was delimited to accommodate Phellinidium asiaticum, Phellinidium ferrugineofuscum (the generic type), Phellinidium fragrans and Phellinidium pouzarii. Accordingly, the concept of Phellinidium was emended to accommodate resupinate species bearing cylindrical to oblong-ellipsoid or allantoid basidiospores. No species of Phellinidium under the new circumscription has been reported to be a forest pathogen. Phellinidium noxium and Phellinidium rufitinctum were excluded from Phellinidium, while the taxonomical positions of Phellinidium aciferum, Phellinidium lamaënse, and Phellinidium orientale are still uncertain., (Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Differentiation of haustoria on primary infection hyphae of the fungal pathogen Puccinia striiformis was studied in wheat seedlings with two-photon microscopy in combination with a classical staining technique. Our results showed a significant increase in the average haustorium size 22, 44, 68, 92 and 116 h after inoculation (hai). After 116 hai no significant change was observed until 336 hai. Haustorium morphology also changed significantly during the time of infection. Initially small spherical haustoria were seen, but as they grew the haustoria gradually became apically branched. At 22 hai all observed haustoria were spherical, but at 44 hai most haustoria had an irregular structure, and at 92 hai all observed haustoria appeared branched. Along with the changes of the haustorial body the haustorial neck changed from narrow and slender to having an expanded appearance with a rough and invaginated structure. The structural changes were similar in two susceptible wheat varieties, 514W and Cartago, although the mean haustorium size was larger in 514W than in Cartago at all intervals.

Molecular distinction between Rhizopogon vinicolor and R. vesiculosus has been made recently, but the diagnostic "yellow-brown (fresh) inflated cells" of R. vesiculosus, originally described by AH Smith, were not observed. These distinctive hyphal cells (vesicles) have not been reported since the type description. In that description they were said to collapse upon drying and described as being difficult to find. Here we report the rediscovery of these vesicles and describe their specific location on sporocarps of R. vesiculosus. We also report an original discovery that coiled, dark-walled hyphae on the sporocarps and in the mycorrhizae of R. vinicolor are of taxonomic value. The coiled hyphae, combined with the presence or absence of the elusive vesicles, allow R. vesiculosus and R. vinicolor to be morphologically distinguished with increased accuracy.

We describe and illustrate two Strobilomyces species, S. seminudus and S. hongoi sp. nov. These two species have been confused and treated as a single species (i.e. S. seminudus). However recent studies based on population genetics have implied that they are reproductively isolated. In the present study we found that they are phylogenetically and morphologically distinct. The molecular phylogenetic trees inferred from the partial sequences of the largest subunit of RNA polymerase II (RPB1) and the second-largest subunit of RNA polymerase II (RPB2) support the differentiation of these two species as well as their differentiation from other related species. Strobilomyces seminudus is characterized by a stipe with an annular zone, becoming distinctly thickened near the apex and mottled with appressed-tomentose scales near the base. In contrast S. hongoi is characterized by a stipe with a remarkable reticulum at the upper and middle part and with minutely warty scales downward. Stipe characteristics also are useful for distinguishing these two species from other related species. In addition the incompletely reticulated basidiospores of these two species are also distinct from those of related species (i.e. S. foveatus).

Here we report the characterization of fungi from 10 accretion ice sections (3300-5100 y old) as well as two deep glacial ice sections that are close to the bottom of the glacier (1,000,000-2,000,000 y old) from the Vostok, Antarctica, 5G ice core. Fungi were characterized by fluorescence microscopy culturing and sequence analyses of ribosomal DNA internal transcribed spacers. A total of 270 fungal colonies were cultured from the accretion ice of subglacial Lake Vostok and an additional 14 from the glacial ice immediately above the accretion ice. Mean concentrations were 0-4.42 cells mL(-1) ice meltwater of which 0-100% exhibited viability (as determined by fluorescence microscopy). Thirty-one unique fungal ribosomal DNA sequences (28 from accretion ice and three from glacial ice) were determined and compared to recent taxa. The results, plus tests for growth at low temperatures, indicated that Lake Vostok contains a mixture of heterotrophic psychrotolerant fungal species. This indicates that the lake is not sterile but contains a unique ecosystem.

Quantitative and qualitative relationships were found between secreted proteins and their activity, and the hyphal morphology of Moniliophthora perniciosa, the causal agent of witches' broom disease in Theobroma cacao. This fungus was grown on fermentable and non-fermentable carbon sources; significant differences in mycelial morphology were observed and correlated with the carbon source. A biological assay performed with Nicotiana tabacum leaves revealed that the necrosis-related activity of extracellular fungal proteins also differed with carbon source. There were clear differences in the type and quantity of the secreted proteins. In addition, the expression of the cacao molecular chaperone BiP increased after treatment with secreted proteins, suggesting a physiological response to the fungus secretome. We suggest that the carbon source-dependent energy metabolism of M. perniciosa results in physiological alterations in protein expression and secretion; these may affect not only M. perniciosa growth, but also its ability to express pathogenicity proteins.

Growth and organelle morphology in the wood rotting basidiomycete fungus Phanerochaete velutina were examined in Petri dishes, on agar-coated slides, and in submerged cultures, using DIC, fluorescence and four-dimensional (4-D; x,y,z,t) confocal microscopy, with several fluorescent probes. Phanerochaete is ideal for this work because of its fast growth, robustness, and use in a wide range of other studies. The probe carboxy-DFFDA, widely used for labelling vacuoles, has no effect either on hyphal tip extension or colony growth at the concentrations usually applied in labelling experiments. Carboxy-DFFDA labels the vacuoles and these form a tubular reticulum in hyphal tip cells. The probe also labels extremely small vesicles (punctate fluorescence) in the apex of tip cells, the Spitzenkörper, and short tubules that undergo sequences of characteristic movements and transformations to produce various morphologies, including ring-like structures. Their location and behaviour suggest that they are a distinct group of structures, possibly a subset of vacuoles, but as yet to be fully identified. Regular incursions of tubules extending from these structures and from the vacuolar reticulum into the apical dome indicate the potential for delivery of material to the apex via tubules as well as vesicles. Such structures are potential candidates for delivering chitin synthases to the apex. Spitzenkörper behaviour has been followed as hyphal tips with linear growth encounter obstacle hyphae and, as the hydrolysis product of carboxy-DFFDA only accumulates in membrane-enclosed compartments, it can be inferred that the labelled structures represent the Spitzenkörper vesicle cloud. Mitochondria also form a reticular continuum of branched tubules in growing hyphal tips, and dual localisation with DiOC6(3) and CMAC allows this to be distinguished from the vacuolar reticulum. Like vacuolar tubules, mitochondrial tubules also span the septa, indicating that they may also be a conduit for intercellular transport.

Fomitiporia punicata sp. nov., collected from Shaanxi Province, northern China, and so far exclusively on Punica granatum (Punicaceae), is described and illustrated, and its preliminary phylogenetic relationships are inferred based on sequence data from the ribosomal ITS-5.8S regions. Fomitiporia punicata belongs to the F. robusta-F. punctata complex, but is characterized by an effused-reflexed to pileate basidioma with triquetrous to ungulate pilei, pale yellowish brown to cinnamon brown pore surface, moderately thick-walled skeletal hyphae in the context, which are partly collapsed on drying, lack of setae, presence of fusoid to subulate cystidioles, and growth on Punica. It also forms a distinct clade within Fomitiporia, and is related to some species from the temperate or Mediterranean zone of the northern hemisphere. An identification key to the species of Fomitiporia recorded in China is provided. Fomitiporia tibetica is transferred to Pseudoinonotus.

A common resupinate thelephoroid fungus was collected in northern Guinean seasonal forests in central and north of Benin (West Africa). The species is reminiscent of Tomentella umbrinospora with respect to the color and thickness of basidiomata and rhizomorphs, the shape of basidiospores in frontal view and the size of subicular hyphae. Both species fall phylogenetically within two clades. Based on detailed anatomical comparison (mostly of rhizomorphs and basidiospores) with the holotype of T. umbrinospora and phylogenetic analyses including ITS rDNA sequences of 40 Tomentella species, T. africana is described as a new species. Genetic distance between the newly described species and T. umbrinospora is 12.1-12.9%, based on ITS rDNA sequences. T. africana is characterized anatomically by yellow-brown thick (0.3-0.8 mm) monomitic rhizomorphs that are commonly covered by irregularly shaped thin hyphae, thin- to thick-walled subicular hyphae of two size ranges, clavate and clamped basidia of 30-60 microm and light yellow to pale brown echinulate basidiospores with irregular shape in frontal view. Detailed anatomical and molecular dissimilarities between T. africana and close species are discussed. Differences between irregularly shaped surface thin hyphae and skeletal ones are highlighted. We stress the relevance of rhizomorphal structures in the discrimination of resupinate thelephoroid fungi.

Aims: To investigate the involvement of oxidative stress and thiol redox state (TRS) in sclerotial differentiation of Sclerotium rolfsii and Sclerotinia sclerotiorum., Methods and Results: Oxidative stress in these fungi was assessed by lipid peroxidation, which was higher in comparison with their nonsclerotiogenic counterpart strains. TRS [measured as glutathione (GSH) and cysteine] was associated with oxidative stress and differentiation using the TRS modulator and antioxidant Nu-acetylcysteine (AcCSH) and the GSH biosynthesis inducer and inhibitor l-2-oxo-thiazolidine-4-carboxylate and L-buthionine-S,R-sulphoximine (BSO) respectively. Differentiation and oxidative stress was decreased by AcCSH in both fungi. The decrease of differentiation by BSO was not associated with oxidative stress in these fungi., Conclusions: Differentiation and oxidative stress in both fungi depends on the availability of antioxidant noncytotoxic -SH groups and is not depended on any direct antioxidant role of GSH and its precursor cysteine., Significance and Impact of the Study: This study helps to understand the mechanism(s) of sclerotial differentiation in these agriculturally important phytopathogenic fungi and proposes that AcCSH can be used as potent fungicide by (i) acting as growth inhibiting cytotoxic oxidant and (ii) sustaining these fungi in their undifferentiated hyphal stage where they are vulnerable to degradation by soil micro-organisms.

The fruit body and in particular the gleba development of several species of Lycoperdaceae has been examined by light microscopic analysis of microtome sections of fruit body primordia in different ontogenetic stages. The gleba of Langermannia gigantea develops in a unique and previously unknown manner: the hymenium-forming palisade structures are borne on fan-like branches of hyphae. Hence the term of flabelloid glebal development is introduced. The other genera of Lycoperdaceae, including the genus Calvatia, are characterized by both lacunar and coralloid development of the gleba. Because these features cannot be distinguished clearly, this type of glebal development is referred to as coralloid-lacunar. Due to the peculiar ontogeny I suggest keeping the genus Langermannia separate from Calvatia.

The phylogenetic relationships of two Japanese Heterobasidion species, H. annosum sensu lato and an undetermined species, were revealed based on three gene loci, glyceraldehyde 3-phosphate dehydrogenase (gpd), heat shock protein (hsp) and elongation factor 1-alpha (ef). The tree, based on combined data of gpd, hsp and ef, showed that Japanese H. annosum s.l. was close to the European S-group, forming a subclade. The results of this study also provided strong support for the recognition of the undetermined Heterobasidion sp. as a distinct phylogenetic species closely related to H. araucariae.

Thirty single basidiospore isolates (SBIs) obtained from four field isolates of the basidiomycete fungus Thanatephorus cucumeris AG 2-2 IV were examined for heterokaryon formation. SBIs of three of four field isolates (Rh509, 92155 and R94) did not produce a tuft of mycelium in the hyphal interaction zone between paired isolates on 2% charcoal agar. Field isolates Rh509, 92155 and R94 indicated no death of interacting mycelium with their progenies on glass slide and microscopic examination. AFLP (amplified fragment length polymorphism) phenotypes of parent and their SBIs were identical. Field isolates Rh509, 92155 and R94 and their SBIs were homothallic. SBIs obtained from field isolate SA-1 were grouped into two mating types (SBI-M1 and SBI-M2), and a tuft of mycelium was formed between paired SBIs-M1 and -M2. SBIs of field isolate SA-1 indicated that no death and death of interacting mycelium were randomly observed. AFLP phenotypes among SBIs of isolate SA-1 were not identical and were also different from their parent isolate. AFLP phenotypes of tuft mycelia produced between heterothallic SBI-M1 and -M2 were heterokaryotic. The mating system of field isolate SA-1 and its SBIs was heterothallic. Both SBIs-M1 and -M2 further produced tuft mycelium with homothallic field isolates and their SBIs. AFLP banding patterns suggested that tuft mycelium was heterokaryotic produced from between heterothallic and homothallic isolates. Results from these experiments clarified that both homothallic and heterothallic isolates exist in population of T. cucumeris AG 2-2 IV, and that genetic exchange can occur between homothallic and heterothallic isolates.

Cord-forming basidiomycetes are important decomposers of dead wood in forest ecosystems but the impact of mycophagous soil invertebrates on their mycelia are little known. Here we investigate the effects of different grazing intensities of Collembola (Folsomia candida) on mycelial foraging patterns of the saprotrophic cord-forming basidiomycetes Hypholoma fasciculare, Phanerochaete velutina and Resinicium bicolor growing from beech (Fagus sylvatica) wood block inocula in dishes of non-sterile soil. Mycelial extension rate and hyphal coverage decreased with increased grazing intensity. R. bicolor was most affected, high grazing density resulting in only a few major cords remaining. Grazing of H. fasciculare often resulted in points of more rapid outgrowth as cords with a fanned margin. In grazed mycelia of P. velutina the main cords had fanned tips and lateral cords became branched. These results suggest that mycophagy by Collembola may hinder the growth of cord-forming fungi in woodlands, which might impact on the ability of these fungi to forage for and decompose dead organic material.

The Spitzenkörper is a dynamic structure present at the tips of hyphal cells with a single highly polarized growth site. It is closely connected with cell morphogenesis and polar growth, and is only present at actively growing sites. Morphogenesis of such highly polarized cells is complex, and requires the coordinated action of multiple protein complexes. We discuss the relevance of these complexes for the structure and function of the Spitzenkörper.

Field ectomycorrhizae sampled under Boletus edulis and Cistus ladanifer have been characterized and described in detail based on standard morphological and anatomical characters. The described ectomycorrhiza has traits typical of Boletales: whitish with three differentiated plectenchymatous layers in the mantle in plan view forming ring-like structures and rhizomorphs with highly differentiated hyphae. The inflated, smooth cystidia-like clavate end cells on the surface of the rhizomorphs and their slightly twisted external hyphae are additional characterizing features. The Hartig net occupies 1 1/2 rows of cortical cells, partly reaching the endodermis. Not all hyphae have clamps. The identification of the fungal symbiont as B. edulis was confirmed by ITS rDNA sequence comparison between mycorrhizas and sporocarps. The singularity of this symbiotic association, as well as its ecological and practical implications, are discussed.

Species in the genus Tomentella (Thelephoraceae) belong to the most frequent and widespread ectomycorrhizal (EM) fungi found in temperate and boreal forests. Although several unidentified tomentelloid morphotypes have been presented as common members of EM communities in coniferous and broad-leaved forests, few tomentelloid EM have been identified and described in detail. In this study, ten tomentelloid EM isolates collected from Populus alba, Quercus cerris and Picea abies stands in Hungary and Germany are characterized and documented by morphological-anatomical methods using light microscopy. The investigated ectomycorrhizae belong to the same brown-black tomentelloid morphotype but form two different anatomotype groups (At I and At II). Molecular taxonomical identification was accomplished using phylogenetic analysis (neighbor joining method) of 49 Tomentella nrDNA-ITS nucleotide sequences including the 10 new and 39 GenBank sequences. The EM isolates clustered into two adjoining clades identical with the two anatomotypes. At II clustered with Tomentella stuposa while At I could not be identified to species. Based on the morphological similarity and the low genetic difference it must be a closely related taxon. A comparison of the recently known tomentelloid EM to T. stuposa is presented. Ecological questions involving abundance and host relationships are discussed.

Auriporia brasilica sp. nov. is described and illustrated based on specimens collected from southern Brazil. It is characterized by a dimitic hyphal system, the shape and size of cystidia, small basidiospores and large pores. This is the first report of an Auriporia taxon registered from the tropics.

The membrane-selective fluorescent dye FM4-64, N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridium dibromide, was used to stain the apical vesicle cluster within the specialized Spitzenkörper of the germ tube of the rust fungi Uromyces vignae and Puccinia graminis f. sp. tritici grown on glass surfaces. The Spitzenkörper stained within 15 min following addition of the dye. Optical sectioning by confocal microscopy of stained hyphal tips showed that the Spitzenkörper was asymmetrically positioned close to the cell-substratum interface during germ tube growth. The Spitzenkörper showed variations in shape and positioning over short (5 s) time intervals. The movement to a new location in the hyphal dome was followed by new growth in that region, consistent with the view that the Spitzenkörper supplies secretory vesicles for germ tube growth. A pronounced Spitzenkörper disappeared at the onset of appressorium differentiation during swelling of the germ tube. However, a stained structure, similar in appearance to a Spitzenkörper, was again observed during the formation of the highly polarized penetration peg.

Tubule formation is a widespread feature of the endomembrane system of eukaryotic cells, serving as an alternative to the better-known transport process of vesicular shuttling. In filamentous fungi, tubule formation by vacuoles is particularly pronounced, but little is known of its regulation. Using the hyphae of the basidiomycete Pisolithus tinctorius as our test system, we have investigated the effects of four drugs whose modulation, in animal cells, of the tubule/vesicle equilibrium is believed to be due to the altered activity of a GTP-binding protein (GTP gamma S, GDP beta S, aluminium fluoride, and Brefeldin A). In Pisolithus tinctorius, GTP gamma S, a non-hydrolysable form of GTP, strongly promoted vacuolar tubule formation in the tip cell and next four cells. The effects of GTP gamma S could be antagonised by pre-treatment of hyphae with GDP beta S, a non-phosphorylatable form of GDP. These results support the idea that a GTP-binding protein plays a regulatory role in vacuolar tubule formation. This could be a dynamin-like GTP-ase, since GTP gamma S-stimulated tubule formation has only been reported previously in cases where a dynamin is involved. Treatment with aluminium fluoride stimulated vacuolar tubule formation at a distance from the tip cell, but NaF controls indicated that this was not a GTP-binding-protein specific effect. Brefeldin A antagonised GTP gamma S, and inhibited tubule formation in the tip cell. Given that Brefeldin A also affects the ER and Golgi bodies of Pisolithus tinctorius, as shown previously, it is not clear yet whether the effects of Brefeldin A on the vacuole system are direct or indirect., (Copyright 2002 Wiley-Liss, Inc.)

Our examination of the cytological characteristics of the vegetative incompatibility reaction in a filamentous basidiomycete, Helicobasidium monpa, by analyzing the fluorescence emitted by ethidium bromide and acridine orange stained nuclei is described. Hyphal anastomoses between strains belonging to different mycelium compatibility groups (MCG) were observed with cell death in fused hyphae, whose nuclei were intensified by ethidium bromide. In contrast, the nuclei in a living cell were not intensified by staining with ethidium bromide, but were intensified by staining with acridine orange. These results indicate that in H. monpa, ethidium bromide staining is a useful method for detecting dead cells. We also examined the relationships between the alternation of ploidy and hyphal anastomosis formation using the newly developed method on filamentous fungi. The tetraploid monokaryon strain derived from the original dikaryon strain by continuous subculture could not be fused to any wild type strains, but the original dikaryon strain could be fused without cell death to only the same MCG strain. In contrast, the haploid dikaryon strain derived from the original monokaryon strain fuses to several strains belonging to different MCGs without cell death. These results suggested that the cellular ploidy of this fungus is closely related to its mating system and, H. monpa may be a self-fertilizing fungus.

The genus Kondoa Y. Yamada, Nakagawa & Banno was erected to accommodate a single taxon, K. malvinella (Fell & Hunter) Y. Yamada, Nakagawa & Banno, which was transferred from the teliospore-forming genus Rhodosporidium Banno based on pronounced differences in the 5S and 26S ribosomal RNA (rRNA) nucleotide sequences to R. toruloides Banno. In contrast with the original description, reinvestigation of K. malvinella revealed the formation of transversely septate (auricularioid) basidia that did not arise on teliospores, but formed directly on the dikaryotic mycelium. The four-celled basidia developed sterigmata on which forcibly discharged asymmetric basidiospores (ballistospores) were produced. Additionally, a new taxon emerged from the study of recent isolates, for which the name K. aeria sp. nov. is proposed. This new species produced two-celled auricularioid basidia on hyphae with incomplete clamp connections. Ballistospores arose on the basidia at the tip of sterigmata and, after ejection, germinated by budding. These observations led us to present an emended diagnosis for the genus Kondoa. Analysis of the sequence data from the D1/D2 region of the 26S rRNA gene showed a very close resemblance between K. aeria and K. malvinella in a cluster that also contained several Bensingtonia species. Taxa in this cluster share specific physiological traits and produce characteristic pinkish-cream to mauve colonies; in contrast, formation of ballistoconidia is only observed in the Bensingtonia species. Sequence data supported placement of K. malvinella and K. aeria in the 'Agaricostilbum clade' of the Urediniomycetes.