Propagule size is not a good predictor for regional population subdivision or fine-scale spatial structure in lichenized fungi.

Propagule size has important consequences on the genetic structure of wind-dispersed species, as species with small propagules have higher capability of long-distance dispersal. Here, we studied reproductive modes and compared local and regional population structures in three Macaronesian lichenized fungi differing in propagule size. First, we quantified size distribution of propagules in each species. Second, genotype simulations based on microsatellite data were used to infer the reproductive mode. Third, using spatial analysis and population genetic approaches, we quantified the local and regional scale genetic structures of the fungal species. The three species differed in size distributions of propagules. The majority of populations exhibited clonal reproductive mode. Identical reproductive modes occurred often across species in the same sites, implying a possible relationship between reproductive mode and local site conditions. Contrary to expectation, at the local scale, the species exhibited similar patterns of spatial autocorrelation in genotypes. However, in agreement with the expectation based on propagule size, the species with highest frequency of small vegetative propagules (L. pulmonaria) exhibited lowest regional genetic differentiation. Nevertheless, altogether, our results show that propagule size is not a good predictor of population subdivision in lichenized fungi, neither at local nor regional spatial scale.

Agar block smear preparation: a novel method of slide preparation for preservation of native fungal structures for microscopic examination and long-term storage.

We describe a novel method of fungal slide preparation named "agar block smear preparation." A total of 510 agar block smears of 25 fungal strains obtained from culture collections, 90 QC fungal strains, and 82 clinical fungal strains from our clinical microbiology laboratory, which included a total of 137 species of yeasts, molds, and thermal dimorphic fungi, were prepared and examined. In contrast to adhesive tape preparation, agar block smears preserved the native fungal structures, such as intact conidiophores of Aspergillus species and arrangements of conidia in Scopulariopsis brevicaulis. Furthermore, agar block smears allowed examination of fungal structures embedded in the agar, such as the ascomata with ascomal hairs in Chaetomium funicola; pycnidium of Phoma glomerata; the intercalary ovoidal chlamydospores arranged in chains of Fusarium dimerum; and the lateral, spherical chlamydospores arranged in pairs of Fusarium solani. After 1 year of storage, morphological integrity was found to have been maintained in 459 (90%) of the 510 agar block smears. After 3 years of storage, morphological integrity was found to have been maintained in 72 (71%) of the 102 smears prepared in 2006. Agar block smear preparation preserves the native fungal structures and allows long-term storage and examination of fungal structures embedded in the agar, hence overcoming the major drawbacks of adhesive tape preparation. The major roles of agar block smear should be diagnosis for difficult cases, accurate identification of fungal species for clinical management of patients and epidemiological studies, and long-term storage for transportation of slides and education purposes.

The structure and histochemistry of sclerotia of Ophiocordyceps sinensis.

The structure and histochemistry of sclerotia of Ophiocordyceps sinensis (synonym: Cordyceps sinensis) are described. The remains of the caterpillar epidermis and sometimes setae of the caterpillar were attached to the pigmented layer that is external to the rind of the sclerotium. The outer aerial hyphae and hyphae of the inner medulla were densely interwoven around the epidermis of the caterpillar; these eventually differentiated into the rind of the sclerotium. The medulla of the sclerotium consisted of three intergrading regions of hyphal density: high, low and a region of intermediate hyphal density. All hyphae of the medulla contained large quantities of protein, polysaccharide and polyphosphate; only the region of high hyphal density was rich in beta-1,3 glucans; the center of the sclerotium was almost devoid of hyphae and contained what are most likely the remains of caterpillar tissue. These features are compared with those of sclerotia of other fungi, and their possible significance is discussed.

Magnaporthe grisea cutinase2 mediates appressorium differentiation and host penetration and is required for full virulence.

The rice blast fungus Magnaporthe grisea infects its host by forming a specialized infection structure, the appressorium, on the plant leaf. The enormous turgor pressure generated within the appressorium drives the emerging penetration peg forcefully through the plant cuticle. Hitherto, the involvement of cutinase(s) in this process has remained unproven. We identified a specific M. grisea cutinase, CUT2, whose expression is dramatically upregulated during appressorium maturation and penetration. The cut2 mutant has reduced extracellular cutin-degrading and Ser esterase activity, when grown on cutin as the sole carbon source, compared with the wild-type strain. The cut2 mutant strain is severely less pathogenic than the wild type or complemented cut2/CUT2 strain on rice (Oryza sativa) and barley (Hordeum vulgare). It displays reduced conidiation and anomalous germling morphology, forming multiple elongated germ tubes and aberrant appressoria on inductive surfaces. We show that Cut2 mediates the formation of the penetration peg but does not play a role in spore or appressorium adhesion, or in appressorial turgor generation. Morphological and pathogenicity defects in the cut2 mutant are fully restored with exogenous application of synthetic cutin monomers, cAMP, 3-isobutyl-1-methylxanthine, and diacylglycerol (DAG). We propose that Cut2 is an upstream activator of cAMP/protein kinase A and DAG/protein kinase C signaling pathways that direct appressorium formation and infectious growth in M. grisea. Cut2 is therefore required for surface sensing leading to correct germling differentiation, penetration, and full virulence in this model fungus.

Powdery mildew (Sphaerotheca fuliginea) resistance in melon is selectable at the haploid level.

The major cause of powdery mildew in melons (Cucumis melo L.) is the fungus Sphaerotheca fuliginea. There are several cultivar- and season-specific races of this fungus. In order to control powdery mildew, it is important to introduce resistance to fungal infection into new cultivars during melon breeding. Haploid breeding is a powerful tool for the production of pure lines. In this study, it was investigated whether powdery mildew resistance could be manifested at the haploid level from two disease-resistant melon lines, PMR 45 and WMR 29. the effects of various races of S. fuliginea on diploid and haploid plants of PMR 45 and WMR 29 and of a disease-susceptible line, Fuyu 3 were measured. The responses of haploid and diploid plants to powdery mildew were identical. In addition, haploids that were generated from hybrids between Fuyu 3 and disease-resistant lines were examined. Seven out of 13 haploids from a Fuyu 3xPMR 45 cross and 10 out of 12 haploids from a Fuyu 3xWMR 29 cross were classified as resistant plants because they showed the same responses as their disease-resistant diploid parents to the various fungal races. These results indicate that resistance in PMR 45 and WMR 29 is selectable at the haploid level. All of the plant responses were observed by microscopy. A possible mechanism for generating powdery mildew resistance in two different melon lines is discussed.

A novel gene, CBP1, encoding a putative extracellular chitin-binding protein, may play an important role in the hydrophobic surface sensing of Magnaporthe grisea during appressorium differentiation.

The conidial germ tube of the rice blast fungus, Magnaporthe grisea, differentiates a specialized cell, an appressorium, required for penetration into the host plant. Formation of the appressorium is also observed on artificial solid substrata such as polycarbonate. A novel emerging germ tube-specific gene, CBP1 (chitin-binding protein), was found in a cDNA subtractive differential library. CBP1 coded for a putative extracellular protein (signal peptide) with two similar chitin-binding domains at both ends of a central domain with homology to fungal chitin deacetylases and with a C-terminus domain rich in Ser/Thr related extracellular matrix protein such as agglutinin. The consensus sequence of the chitin-binding domain found in CBP1 has never been reported in fungi and is similar to the chitin-binding motif in plant lectins and plant chitinases classes I and IV. CBPI was disrupted in order to identify its function. Null mutants of CBP1 failed to differentiate appressoria normally on artificial surface but succeeded in normally differentiating appressoria on the plant leaf surface. Since the null mutant Cbp1- showed abnormal appressorium differentiation only on artificial surfaces and was sensitive to the chemical inducers, CBP1 seemed to play an important role in the recognition of physical factors on solid surfaces.

Microfungi on compositae in the Ruhr Basin.

Forty-three microfungi have been observed on thirty species of the Compositae occurring in several locations in the Ruhr Basin in North Rhine-Westphalia. Many fungi belong to the Ascomycetes (Erysiphales, Diaporthales, Dothideales, Leotiales and Pleosporales) and to the Deuteromycetes (Melanconiales, Moniliales and Sphaeropsidales). Other fungi wich were found in our investigation belong to the Basidiomycetes (Uredinales) and to the Oomycetes (Peronosporales). Some recorded microfungi have been discovered on new hosts in North Rhine-Westphalia and also in Germany for the first time. New for North Rhine-Westphalia are Ascochyta spec. Libert on Matricaria recutita L., Phoma exigua var. linicola (Naumov & Vassilevski) Maas on Tripleurospermum maritimum (L.) W. D. J. Koch, Phomopsis achillea (Sacc.) Höhn. on Achillea ageratum L., Diaporthe aff. arctii (Lasch) Nitschke on Solidago canadensis L. and on Achillea ageratum L., Lophiostoma caulium (Fr.) Ces. & De Not. on Anthemis tinctoria L. and Ophiobolus fructicum (Rob. ex Desm.) on Serratula tinctoria L. New for Germany are Ophiobolus cirsii (P. Karst.) Sacc. on Cichorium intybus L., Phomopsis cirsii Grove on Cirsium vulgare (Savi) Ten., Pleospora kansensis J. P. Ellis & M. B. Ellis and Pleospora phaeocomoides cf. var. infectoria on Centaurea jacea L.

Surveys of microfungi in a former industrial area in Duisburg-Nord.

One hundred and forty microfungi (Ascomycetes and Deuteromycetes) were collected in the "Landschaftspark Duisburg-Nord" located in North Rhine-Westphalia. New hosts for rarely found fungi are recorded for the first time. New for Germany are Massaria inquinans (Tode) De Not. and Nitschkia grevillei (Rhem) Nannf. on Acer pseudoplatanus L., Pirottaea nigrostriata Graddon on Artemisia vulgaris L., Ceratopycnis clematidis Höhn. on Clematis vitalba L., Dasyscyphus aff. humuli (W. Phillips) Dennis on Humulus lupulus L. and Leptosphaeria derasa (Berk. & Br.) Auersw. on Senecio inaequidens DC. New for North Rhine-Westphalia are Chaetosphaerella phaeostroma (Durieu & Mont.) E. Müller & Booth and Phomopsis platanoides (Cooke) Died. on Acer pseudoplatanus L., Microsphaeropsis pseudaspera Sutton, Mycosphaerella osborniae D. Hawksw. & Sivan. and Phomopsis oblita Sacc. on Artemisia vulgaris L., Leptosphaeria acuta (Fr.) P. Karst. and Leptosphaeria doliolum (Pers.) Ces. & De Not. on Bryonia dioica Jacq., Ophiobolus erythrosporus (Riess) G. Winter and Pleospora herbarum (Pers.) Rabenh. ex Ces. & De Not. on Dipsacus sylvestris (Huds), Keissleriella ocellata (Niessl) Bose on Hypericum perforatum L., Dactylaria aff. graminicola on Lolium perenne L., Siroplacodium aff. atrum on Oenothera beinnis L., Diatrypella favacea (Fr.) Sacc. on Prunus spec., Hapalosphaeria deformans (Syd.) Syd. and Microdiscula rubicola (Bres.) Höhn. on Rubus fructicosus agg. L., Cryptodiaporthe salicina (Pers.) Wehm. on Salix alba L. and Pleurophoma pleurospora (Sacc.) Höhn. on Salix caprea L.