A Multilocus Phylogeny Places Hymenula cerealis (syn. Cephalosporium gramineum) in the Helotiales, Leotiomycetes.

The soilborne fungus Hymenula cerealis causes Cephalosporium stripe, a vascular wilt disease of wheat and other grasses in the United States and other wheat-producing countries where winter wheat is subjected to snow cover and frozen soil. No sexual stage is known for H. cerealis, and consequently, its phylogenetic position relative to other fungi has been difficult to establish. The purpose of this study was to conduct a multilocus sequence analysis to determine the phylogenetic position of H. cerealis. Sequence data for five genes, the internal transcribed spacer (ITS), partial large subunit nrDNA (LSU), partial RNA polymerase II second largest subunit region (RPB2), ß-tubulin gene, and translation elongation factor 1-α (TEF1-α), from a diverse set of C. gramineum isolates and other related fungi was obtained from GenBank or directly from isolates in the Murray lab and used to construct maximum-likelihood and Bayesian trees. Based on phylogenetic analysis of the single LSU and ß-tubulin genes, Cephalosporium gramineum is closely related to the Drepanopezizaceae and Ploettnerulaceae of Helotiales. Based on analyses of the DNA sequence of the ITS, RPB2, and TEF1-α genes, as well as the combined five-gene data set, C. gramineum belongs to the family Drepanopezizaceae, which is a sister taxon to the Ploettnerulaceae, and formed a well-supported clade (MLBP/BIPP = 95%/100%). In conclusion, H. cerealis belongs to the Helotiales, Leotiomycetes.

Production of the antibiotic secondary metabolite solanapyrone A by the fungal plant pathogen Ascochyta rabiei during fruiting body formation in saprobic growth.

Fungi are noted producers of a diverse array of secondary metabolites, many of which are of pharmacological importance. However, the biological roles of the vast majority of these molecules during the fungal life cycle in nature remain elusive. Solanapyrones are polyketide-derived secondary metabolites produced by diverse fungal species including the plant pathogen Ascochyta rabiei. This molecule was originally thought to function as a phytotoxin facilitating pathogenesis of A. rabiei. Chemical profiling and gene expression studies showed that solanapyrone A was specifically produced during saprobic, but not parasitic growth of A. rabiei. Expression of the gene encoding the final enzymatic step in solanapyrone biosynthesis was specifically associated with development of the asexual fruiting bodies of the fungus on certain substrates. In confrontation assays with saprobic fungi that were commonly found in chickpea debris in fields, A. rabiei effectively suppressed the growth of all competing fungi, such as Alternaria, Epicoccum and Ulocladium species. Solanapyrone A was directly detected in the inhibitory zone using a MALDI-imaging mass spectrometry, and the purified compound showed significant antifungal activities against the potential saprobic competitors. These results suggest that solanapyrone A plays an important role for competition and presumably the survival of the fungus.

A Novel Penicillium sp. Causes Rot in Stored Sugar Beet Roots in Idaho.

Penicillium vulpinum along with a number of other fungi can lead to rot of stored sugar beet roots. However, Penicillium isolates associated with necrotic lesions on roots from a recent sugar beet storage study were determined to be different from P. vulpinum and other recognized Penicillium species. Phylogenies based on sequencing of the internal transcribed spacer (ITS)-5.8S, ß-tubulin (BenA), and RNA polymerase II second largest subunit (RPB2) DNA regions indicate that these isolates are novel, but most closely related to the following Penicillium spp. in the section Fasiculata: P. aurantiogriseum, P. camemberti, and P. freii. Macro- and micromorphological data also support designating these isolates as a new species for which we propose the name, Penicillium cellarum sp. nov. Inoculation studies with the P. cellarum isolates on roots of the commercial sugar beet cultivar B-7 led to the formation of necrotic lesions 23 to 25 mm in diameter after 86 days in storage. These lesions were similar to those observed on sugar beet roots in commercial storage piles. These data indicate that P. cellarum is a pathogen which can cause root rot in stored sugar beet roots.

Neocosmospora sp.-derived resorcylic acid lactones with in vitro binding affinity for human opioid and cannabinoid receptors.

Bioassay-guided fractionation of a fungus Neocosmospora sp. (UM-031509) resulted in the isolation of three new resorcylic acid lactones, neocosmosin A (2), neocosmosin B (3), and neocosmosin C (4). Three known resorcylic acid lactones, monocillin IV (1), monocillin II (5), and radicicol (6), were also isolated and identified. The structures of these compounds were established on the basis of extensive 1D and 2D NMR spectroscopic analysis, mass spectrometric (ESIMS) data, and X-ray crystallography. Compounds 4-6 show good binding affinity for the human opioid receptors. These findings have important implications for evaluating the potential psychoactive effects with this class of compounds.

Antimicrobial and antiprotozoal activities of secondary metabolites from the fungus Eurotium repens.

In this study, we examined in vitro antibacterial, antifungal, antimalarial, and antileishmanial activities of secondary metabolites (1-8) isolated from the fungus Eurotium repens. All compounds showed mild to moderate antibacterial or antifungal or both activities except 7. The activity of compound 6 was the best of the group tested. The in vitro antimalarial evaluation of these compounds revealed that compounds 1-3, 5, and 6 showed antimalarial activities against both chloroquine-sensitive and chloroquine-resistant strains of Plasmodium falciparum with IC(50) values in the range of 1.1-3.0 µg/ml without showing any cytotoxicity to the mammalian cells. Compound 5 displayed the highest antimalarial activity. Antileishmanial activity against Leishmania donovani promastigotes was observed for compounds 1-6 with IC(50) values ranging from 6.2 to 23 µg/ml. Antileishmanial activity of compounds 5 and 6 (IC(50) values of 7.5 and 6.2 µg/ml, respectively) was more potent than 1-4 (IC(50) values ranging from 19-23 µg/ml). Compounds 7 and 8 did not show any antiprotozoal effect. Preliminary structure and activity relationship studies indicated that antibacterial, antifungal, antimalarial, and antileishmanial activities associated with phenol derivates (1-6) seem to be dependent on the number of double bonds in the side chain, which would be important for lead optimization in the future.

Benzyl derivatives with in vitro binding affinity for human opioid and cannabinoid receptors from the fungus Eurotium repens.

Bioassay-guided fractionation of the fungus Eurotium repens resulted in the isolation of two new benzyl derivatives, (E)-2-(hept-1-enyl)-3-(hydroxymethyl)-5-(3-methylbut-2-enyl)benzene-1,4-diol (1) and (E)-4-(hept-1-enyl)-7-(3-methylbut-2-enyl)-2,3-dihydrobenzofuran-2,5-diol (2), along with seven known compounds (3-9) including five benzaldehyde compounds, flavoglaucin (3), tetrahydroauroglaucin (4), dihydroauroglaucin (5), auroglaucin (6), and 2-(2',3-epoxy-1',3'- heptadienyl)-6-hydroxy-5-(3-methyl-2-butenyl)benzaldehyde (7), one diketopiperazine alkaloid, echinulin (8), and 5,7-dihydroxy-4-methylphthalide (9). The chemical structures of these compounds were established on the basis of extensive 1D and 2D NMR and HRMS data. Compounds 1-4 and 6 showed good binding affinity for human opioid or cannabinoid receptors. These findings have important implications for psychoactive studies with this class of compounds.

Erysiphe trifolii Causing Powdery Mildew of Lentil (Lens culinaris).

The taxonomy of the powdery mildew fungus infecting lentil in the Pacific Northwest (PNW) of the United States was investigated on the basis of morphology and rDNA internal transcribed spacer (ITS) sequences. Anamorphic characters were in close agreement with descriptions of Erysiphe trifolii. However, teleomorphs formed chasmothecial appendages with highly branched apices, whereas E. trifolii has been described as producing flexuous or sometimes loosely branched appendages. Branched appendages have been described in Erysiphe diffusa, a fungus reported from species of Lens, Glycine, and Sophora, raising the possibility that the PNW fungus could be E. diffusa. Examination of morphological characters of an authentic specimen of E. trifolii from Austria determined that it included chasmothecial appendages resembling those seen in PNW specimens. Furthermore, ITS sequences from five powdery mildew samples collected from lentils in PNW greenhouses and fields from 2006 to 2008 were identical to one another, and exhibited higher similarity to sequences of E. trifolii (99%) than to those of any other Erysiphe spp. available in GenBank. Parsimony analysis grouped the lentil powdery mildew into a clade with Erysiphe baeumleri, E. trifolii, and E. trifolii-like Oidium sp., but indicated a more distant relationship to E. diffusa. In greenhouse inoculation studies, the lentil powdery mildew fungus did not infect soybean genotypes known to be susceptible to E. diffusa. The pathogenicity of E. trifolii on lentil was confirmed using modified Koch's postulates. This is the first report of E. trifolii infecting lentil. E. diffusa and E. trifolii have different host ranges, so the discovery of E. trifolii on lentil has implications both for determining species of powdery mildews on cool-season grain legumes, and in disease management.

Didymella pisi sp. nov., the teleomorph of Ascochyta pisi.

The anamorphic pycnidial fungus Ascochyta pisi is one member of a species complex that causes Ascochyta blight of pea, a potentially devastating disease. The teleomorphic state of this fungus was induced under laboratory conditions. Using morphological and molecular characters, we placed the teleomorph within the genus Didymella as D. pisi and describe a heterothallic mating system using a PCR-based mating type assay and in vitro crosses. We compare D. pisi with other Didymella spp. with which it might be confused.

Evaluation of the potential role of water in spread of conidia of the Neotyphodium endophyte of Poa ampla.

Neotyphodium endophytes are asexual, filamentous fungi, mutualistically associated with diverse cool season grasses. Infected seeds and vegetative organs of infected host plants are the only known modes of propagation of the asexual endophytes. In the last decade certain Epichloë and Neotyphodium-infected grass species have been shown to have epiphyllous structures of the endophytes, hyphae, conidiophores, and conidia, growing on leaf blades. The production of epiphyllous conidia suggests the possibility that some of these endophytes may have the ability for plant-to-plant spread. The objective of this study was to determine the possible mechanisms involved in liberation and dispersal of asexual spores of Neotyphodium growing in vitro and epiphyllously on leaves of Poa ampla. Our results indicate that water dispersal is the most likely means of dissemination of conidia of the Neotyphodium sp. Wind generated by dry compressed air does not dislodge the conidia from slide cultures or from P. ampla leaves.