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PubMed2006

Marine fungal lineages in the Hypocreomycetidae.

Phylogenetic analyses of DNA sequences from protein coding and ribosomal nuclear loci support the placement of a number of marine fungal species associated with a well-supported clade containing fungicolous species of Melanospora and wood inhabiting Coronophorales. Three subclades containing marine species were recovered including Torpedospora radiata plus T. ambispinosa, Swampomyces species plus Juncigena adarca, and two Etheirophora species plus additional Swampomyces species. The monophyly of these three subclades, as well as a subclade containing representatives of Coronophorales and Melanospora, is well supported statistically. However, relationships among the different subclades remain largely unresolved. A sister relationship for this group with Hypocreales is significantly supported by Bayesian and ML methods. In addition to the Halospheariales and Lulworthiales, this clade, which is characterized by considerable morphological and ecological diversity, represents a third major clade of marine Sordariomycetes.

Ascomycota↗

PubMed2003

Molecular detection of ascomycetes associated with Fucus serratus.

The association of ascomycetes with Fucus serratus was investigated by comparing the broad-based molecular and cultural diversities of healthy and dead fronds. Four PCR primer pairs were used to amplify the 18S (primers NS1-FR1; NS1-EF3) or 28S rRNA (primers NL209-NL912; NL359-NL912) genes directly from the DNA of algal thalli. Two novel primer pairs, NL209-NL912 and NL359-NL912 giving product sizes of 700 and 559 bp respectively, were designed to amplify the 28S rDNA from ascomycetes specifically. All primer combinations amplified DNA from 33 reference taxa isolated from Fucus serratus, and the products generated by primers NS1-FR1GC and NL359-NL912GC were separated in 18-38% and 38-60% denaturant gradients respectively after DGGE. The 18S rDNA DGGE system resolved eight bands from algal DNA, but many of the sequences separated were not fungal, whereas the 28S rDNA system resolved seven bands that were all identified as ascomycetes. Phylogenetic analysis and BLAST search results of environmental sequences revealed the presence of four main ascomycete groups: (1) the Halosphaeriales, (2) the Hypocreales, (3) an unidentified Lulworthiales complex, and (4) the Pleosporales. Few fungal isolates were detected molecularly suggesting that fungal colonisation of fronds was limited, mainly to species in dead casts.

Ascomycota↗

PubMed2004

Conspecificity of the cerulenin and helvolic acid producing 'Cephalosporium caerulens', and the hypocrealean fungus Sarocladium oryzae.

Fermentation processes for the biochemical reagents cerulenin and helvolic acid employ 'Cephalosporium caerulens,' an invalidly published designation that has been used for more than 40 years. However, its identity has never been critically examined because strains were unavailable from major culture collections. An authentic strain of C. caerulens', derived from the original strain KF-140, was recently found and compared to Sarocladium oryzae, another Acremonium-like fungus which also produces cerulenin and helvolic acid. Morphological comparisons, rDNA sequence data, and chromatography of secondary metabolites established that 'C. caerulens' and S. oryzae are conspecific. Sequence data from ribosomal DNA genes indicated S. oryzae belongs to the Hypocreales and is allied with members of the Ceratostomataceae, Scopinella species, Emericellopsis species and certain Acremonium-like anamorphs of uncertain familial relationships. At least two of the isolates of S. oryzae produced titres of cerulenin and helvolic acid similar to those of KF-140. This finding demonstrates that manufacture of cerulenin need not be limited to the original strain.

Acremonium↗

PubMed2005

On the relationships of Paecilomyces sect. Isarioidea species.

Phylogenetic relationships of Paecilomyces sect. Isarioidea species were analysed using the beta-tubulin gene and ITS rDNA. Maximum parsimony analyses showed that the section does not form a natural taxonomic group and is polyphyletic within the Hypocreales. However, a group was recognized, designated as the Isaria clade, to be monophyletic comprising of the following Paecilomyces species: P. amoeneroseus, P. cateniannulatus, P. cateniobliquus, P. cicadae, P. coleopterorus, P. farinosus, P. fumosoroseus, P. ghanensis, P. javanicus and P. tenuipes. Some of these species have teleomorphs in Cordyceps.

Cordyceps↗

PubMed2005

Molecular and immunochemical phylogeny of Verticillium species.

21 strains with all typical morphological characteristics of eight Verticillium species (Phyllachorales) were studied in this work, together with representatives from four Hypocreales species (11 strains), that were previously classified as members of the genus. The PCR products from two nuclear genes, i.e. the ITS1-5.8S-ITS2 region and RNA polymerase II largest subunit gene (rpb1), together with four mitochondrial genes, i.e. the small ribosomal rRNA subunit (rns), the two NADH dehydrogenase subunit genes (nad1 and nad3), and the cytochrome oxidase subunit III gene (cox3) were sequenced and analyzed. Similarly, antibodies raised against one strain of each of the species examined (V. nubilum and V. theobromae excluded) were used against the proteins of all other strains. The number and relative area of precipitates formed after crossed electrophoreses served to estimate the degree of immunochemical relatedness. Combined molecular and immunochemical data clarified the phylogenetic relationships of all true Verticillium species and provided a convincing insight into the evolutionary relation of the sect. Nigrescentia with members of the sect. Verticillium and sect. Prostrata that until recently were included in Verticillium.

Cytochromes↗

PubMed2000

Screening of basidiomycetes for antimicrobial activities.

As a part of a screening programme developed to evaluate the antimicrobial activity of basidiomycetes, 317 isolates representing 204 species collected in Spain were screened against a range of human clinical pathogens and laboratory controls. Extracts from 45% of the isolates, representing 109 species, showed antimicrobial activity. Antibacterial activity was more pronounced than antifungal activity. The proportion of extracts from basidiomycetes showing antimicrobial activity was similar to or above that obtained for representative orders of Ascomycetes, such as Pezizales and Xylariales, but lower than that produced by members of the orders Diaporthales, Eurotiales, Hypocreales, Leotiales and Sordariales. Suprageneric taxa (orders and families) did not show pronounced differences in their antimicrobial activities though such differences were observed at the genus level, suggesting that the ability to produce these bioactive compounds is not homogenously distributed amongst the basidiomycetes. Isolates from some species showed large differences in their ability to produce metabolites with antimicrobial activity, possibly reflecting genetic differences at the infraspecific level.

Anti-Bacterial Agents↗

PubMed2002

Phylogenetic relationships among strains of the entomopathogenic fungus, Nomuraea rileyi, as revealed by partial beta-tubulin sequences and inter-simple sequence repeat (ISSR) analysis.

AIMS: To elucidate the phyletic relationships among three members of the entomogenous fungal genus, Nomuraea, with an emphasis on N. rileyi. METHODS AND RESULTS: Relationships were evaluated by analysis of the beta-tubulin gene and of inter-simple sequence repeats (ISSR). The amplification product of the partial beta-tubulin gene was larger for N. atypicola than for N. rileyi, and sequencing of this gene fragment confirmed that N. atypicola possesses approximately 25 more nucleotides than N. rileyi and N. anemonoides. Based on neighbor joining and bootstrap analysis of the partial beta-tubulin gene, N. atypicola failed to form a monophyletic grouping with the other two species of Nomuraea. In contrast, the single isolate of N. anemonoides clustered with the N. rileyi isolates, and both taxa grouped with Epichloe typhina (Hypocreales: Clavicipitaceae). Results from this study suggested that N. rileyi and N. anemonoides are closely related to the Clavicipitaceae. In contrast, evidence indicated that N. atypicola is not closely related to this family, and that this taxon is not a Nomuraea. Based on the 83 polymorphic loci of ISSR, it was observed that isolates of N. rileyi from diverse geographical origins were distinctly different from both N. atypicola and N. anemonoides. Considerable heterogeneity was observed in the 18 isolates of N. rileyi tested, and several clusters contained isolates from disparate geographical locations and hosts. However, three isolates from the Philippines (three host species) and three strains isolated from velvetbean caterpillar (Anticarsia gemmatalis) larvae in South America did cluster together. Two other strains from Brazil (isolated from Spodoptera spp.) were distinct from the velvetbean caterpillar isolates from South America. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The beta-tubulin gene was generally too conserved to resolve intraspecies variability. However, ISSR did identify polymorphisms among the isolates of N. rileyi tested. The results of this study indicate that ISSR may be used as robust molecular markers for studying the population genetics of this entomopathogenic fungus.

Mitosporic Fungi↗

PubMed1999

The agricultural pathology of ant fungus gardens.

Gardens of fungus-growing ants (Formicidae: Attini) traditionally have been thought to be free of microbial parasites, with the fungal mutualist maintained in nearly pure "monocultures." We conducted extensive isolations of "alien" (nonmutualistic) fungi from ant gardens of a phylogenetically representative collection of attine ants. Contrary to the long-standing assumption that gardens are maintained free of microbial pathogens and parasites, they are in fact host to specialized parasites that are only known from attine gardens and that are found in most attine nests. These specialized garden parasites, belonging to the microfungus genus Escovopsis (Ascomycota: anamorphic Hypocreales), are horizontally transmitted between colonies. Consistent with theory of virulence evolution under this mode of pathogen transmission, Escovopsis is highly virulent and has the potential for rapid devastation of ant gardens, leading to colony mortality. The specialized parasite Escovopsis is more prevalent in gardens of the more derived ant lineages than in gardens of the more "primitive" (basal) ant lineages. Because fungal cultivars of derived attine lineages are asexual clones of apparently ancient origin whereas cultivars of primitive ant lineages were domesticated relatively recently from free-living sexual stocks, the increased virulence of pathogens associated with ancient asexual cultivars suggests an evolutionary cost to cultivar clonality, perhaps resulting from slower evolutionary rates of cultivars in the coevolutionary race with their pathogens.

Journal Article↗

PubMed2005

A new method to evaluate the biocontrol potential of single spore isolates of fungal entomopathogens.

Fifty Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) strains isolated from the coffee berry borer were used to develop a novel screening method aimed at selecting strains with the highest biocontrol potential. The screening method is based on percent insect mortality, average survival time, mortality distribution, percent spore germination, fungal life cycle duration, and spore production on the insect. Based on these parameters, only 11 strains merited further study. The use of a sound scientific protocol for the selection of promising fungal entomopathogens should lead to more efficient use of time, labor, and financial resources in biological control programs.

Animals↗

PubMed2001

Comparison of cell-wall polysaccharides from Nectria cinnabarina with those from the group of Nectria with Sesquicillium anamorphs.

Alkali-extractable and water-soluble polysaccharides were purified from cell walls of five species of Sesquicillium or its teleomorphs, Nectria lasiacidis and Nectria impariphialis, and from Nectria cinnabarina, the type species of Nectria, a heterogeneous genus that belongs to the Hypocreales. Methylation and NMR analyses for determination of linkage types and structure were performed and indicated differences between the polysaccharides purified during the present study and those isolated from other nectrioid fungi, namely the presence of 5-O-substituted galactofuranose (-->5)-Galf-(1-->) in the main chain together with 2,6-di-O-substituted galactofuranose (-->2,6)-Galf-(1-->) residues in Sesquicillium buxi and Sesquicillium pseudosetosum. The polysaccharide from N. impariphialis was similar to those obtained from the above species, although an additional residue of 6-O-substituted glucopyranose (-->6)-Glcp-(1-->), was detected in some side chains. In N. lasiacidis and Sesquicillium candelabrum the polysaccharide contained an additional branching point of 5,6-di-O-substituted galactofuranose (-->5,6)-Galf-(1-->) linked to terminal N-acetylglucosamine GlcNAc-(1-->). These chains were linked to a small mannan core. All these polysaccharides showed major differences to the polysaccharide of N. cinnabarina, which was formed by a main chain of (1-->6)-beta-linked galactofuranose units almost fully branched at positions 2-O by either single residues of glucopyranose or acidic chains containing glucuronic acid and mannose.

Ascomycota↗

PubMed1999

A morphological and molecular perspective of Trichoderma viride: is it one or two species?

Trichoderma (Ascomycetes, Hypocreales) strains that have warted conidia are traditionally identified as T. viride, the type species of Trichoderma. However, two morphologically distinct types of conidial warts (I and II) have been found. Because each type corresponds to a unique mitochondrial DNA pattern, it has been questioned whether T. viride comprises more than one species. Combined molecular data (sequences of the internal transcribed spacer 1 [ITS-1] and ITS-2 regions and of part of the 28S rRNA gene along with results of restriction fragment length polymorphism analysis of the endochitinase gene and PCR fingerprinting), morphology, physiology, and colony characteristics distinguish type I and type II as different species. Type I corresponds to "true" T. viride, the anamorph of Hypocrea rufa. Type II represents a new species, T. asperellum, which is, in terms of molecular characteristics, close to the neotype of T. hamatum.

Chitinases↗

PubMed2004

Detection and quantification of airborne conidia of Fusarium circinatum, the causal agent of pine pitch canker, from two California sites by using a real-time PCR approach combined with a simple spore trapping method.

Pinus radiata (Monterey pine), a tree native to coastal California and Mexico, is widely planted worldwide for timber production. A major threat to Monterey pine plantations is the fungal disease pine pitch canker, caused by Fusarium circinatum (Hypocreales). We present a novel trapping approach using filter paper in combination with a rapid molecular method to detect the presence of inoculum in the air. The assay is also useful for diagnosing the presence of the pathogen on plants. The test is based on the F. circinatum specific primer pair CIRC1A-CIRC4A, which amplifies a 360-bp DNA fragment in the intergenic spacer region of the nuclear ribosomal operon. Real-time PCR was used to calculate the number of fungal spores present in each reaction mixture by comparing the threshold cycle (Ct) of unknown spore samples to the Ct values of standards with known amounts of F. circinatum spores. The filter paper method allows prolonged and more sensitive spore sampling in the field compared to traditional traps using petri dishes filled with selective medium. A field test at two sites in coastal California infested with pine pitch canker was carried out during the summer and fall of 2002. Spore counts were in the range of ca. 1 x 10(3) to ca. 7 x 10(5)/m(2), with the highest spore counts in the fall, suggesting a seasonal fluctuation.

Air Microbiology↗

PubMed1999

Developments in fungal taxonomy.

Fungal infections, especially those caused by opportunistic species, have become substantially more common in recent decades. Numerous species cause human infections, and several new human pathogens are discovered yearly. This situation has created an increasing interest in fungal taxonomy and has led to the development of new methods and approaches to fungal biosystematics which have promoted important practical advances in identification procedures. However, the significance of some data provided by the new approaches is still unclear, and results drawn from such studies may even increase nomenclatural confusion. Analyses of rRNA and rDNA sequences constitute an important complement of the morphological criteria needed to allow clinical fungi to be more easily identified and placed on a single phylogenetic tree. Most of the pathogenic fungi so far described belong to the kingdom Fungi; two belong to the kingdom Chromista. Within the Fungi, they are distributed in three phyla and in 15 orders (Pneumocystidales, Saccharomycetales, Dothideales, Sordariales, Onygenales, Eurotiales, Hypocreales, Ophiostomatales, Microascales, Tremellales, Poriales, Stereales, Agaricales, Schizophyllales, and Ustilaginales).

Ascomycota↗

PubMedDate not supplied

Clarification of the host substrate of Ascopolyporus and description of Ascopolyporus philodendrus sp. nov.

During a recent collection trip to Barro Colorado Island, Panama, two species belonging to genus Ascopolyporus (Clavicipitaceae, Hypocreales) were collected. Species of Ascopolyporus are epibionts of their bamboo (Poaceae) host and long thought to be biotrophs of their plant hosts. However, based on morphological observations and phylogenetic evidence using large subunit ribosomal DNA data, we propose that genus Ascopolyporus is likely composed of pathogens of scale insects (Coccoideae, Homoptera). Phylogenetic analyses included Ascopolyporus spp. in a clade containing only entomopathogenic clavicipitaceous species (100% posterior probability), and the scale insect pathogen Hyperdermium bertonii was found to share the most recent common ancestor with the Ascopolyporus clade (98% posterior probability). In addition remnants of the scale insect were observed to be embedded within stromata during early stages of stroma development. Ascopolyporus philodendrus sp. nov. was described and distinguished from the type species of the genus, A. polychrous, based on perithecial size, ascus size, plant host substrate and phylogenetic evidence. Furthermore subfamily Clavicipitoideae (Clavicipitaceae) was included and well supported in a single clade (100% posterior probability).

Ascomycota↗

PubMedDate not supplied

Hypocrea voglmayrii sp. nov. from the Austrian Alps represents a new phylogenetic clade in Hypocrea/Trichoderma.

The holomorph of the new species Hypocrea voglmayrii (Hypocreales, Ascomycota, Fungi) is described by a combined approach, using morphology of the teleomorph, morphology of the anamorph, culture studies and phylogenetic analyses of ITS1 and 2, ech42 and rpb2 gene sequences. Its anamorph Trichoderma voglmayrii is described as a new anamorph species. Unlike most other species of Hypocrea the teleomorph of H. voglmayrii occurs on dry standing trunks and exhibits well defined black ostioles. Although exclusively collected at higher altitudes, this species grows at 35 C in culture. Hypocrea voglmayrii develops pale yellowish to greenish conidia, a yellowish pigment and a coconut-like odor on CMD. Phylogenetically, H. voglmayrii forms a distinct, isolated branch between the section Trichoderma and the H. pachybasioides clade but does not associate with any of these clades in different gene trees.

Austria↗

PubMedDate not supplied

Hypocrea crystalligena sp. nov., a common European species with a white-spored Trichoderma anamorph.

The new species Hypocrea crystalligena (Hypocreales, Ascomycota, Fungi) is described as a holomorph and characterized based on an integrated phenotypic and phylogenetic approach, using teleomorph and anamorph morphologies, culture studies and analyses of phylogenetic markers including internal transcribed spacer 1 and 2 (ITS1 and 2), two last introns of the translation elongation factor 1-alpha encoding gene (tef1), and a portion of the rpb2 gene, encoding the second largest RNA polymerase subunit. Stromata of H. crystalligena show similarities with those of species from Trichoderma sect. Trichoderma but differ in several respects, including color, presence of white crystals on the surface and small ascospores. Colonies on CMD appear distinct, form colorless to white crystals on isolation, a yellowish to brown pigment and an anamorph with hyaline conidia exhibiting verticillium-like to gliocladium-like structural elements. ITS1 and 2 sequences exhibit all genus-specific features but also contain several unique hallmarks permitting development of a species-diagnostic barcode. Based on the analyses of partial rpb2 and tef1 sequences, H. crystalligena constitutes a separate evolutionary lineage with H. megalocitrina and H. psychrophila as its nearest neighbors. All these species form one phylogenetic clade with the H. pulvinata/H. citrina node.

DNA, Fungal↗

PubMed2001

Microfungi on Rosaceae in eastern Westphalia (Sauerland, North Rhine-Westphalia, Germany).

Microfungi occuring on Rosaceae in the Sauerland were examined. 38 microfungi from 27 different genera were found, including Ascomycetes of the orders Erysiphales, Helotiales, Hypocreales and Taphrinales and Basidiomycetes of the order Uredinales as well as mitosporic fungi. The examined samples have been collected in the period of march to september 2000. New hosts could be found for the following species: Discosia potentillae Lib., Phragmidium acuminatum (Fr.) Cooke, Stigmina carpophila (Lev.) M.B. Ellis, Seimatosporium pestalozzioides (Sacc.) B. Sutton, Septocyta ruborum (Lib.) Petr. and Zythiostroma mougeotii (Fr.) Höhn. Discosia potentillae Lib. is recorded for the first time for Germany and Cercospora potentillae Chupp & Greene for the first time for Europe.

Fungi↗

PubMed2000

Antioxidant activity of the extracts from fruiting bodies of cultured Cordyceps sinensis.

Cordyceps sinensis is one of the most valued herbs in traditional Chinese medicine. We investigated the antioxidant activities of the cultured fruiting bodies of Cordyceps sinesis. The water and ethanol extracts of Cordyceps sinensis were found to possess a potent antioxidant activity. The scavenging effects of the extracts on superoxide were very weak, but the extracts moderately inhibited malondialdehyde formation via hydroxyl radical induced by SIN-1, a peroxynitrite generator. Of the extracts examined, the hot water extract (70 degrees C for 5 min) showed the greatest oxygen free radical scavenging activity. Also, when low-density lipoprotein (LDL) was incubated with macrophages in the presence of CuCl2 (1 microM), the hot water extract showed a strong inhibitory effect against lipid peroxidation in the medium and consequent accumulation of cholesteryl ester in macrophages. Their activities were comparable to that of authentic Cu/Zn SOD. These results suggest that the extracts of cultured Cordyceps sinensis possess potent antioxidant and anti-lipid peroxidation activities and inhibit accumulation of cholesteryl ester in macrophages via suppression of LDL oxidation.

Animals↗