Reassessment of type specimens of Cordyceps and its allies described by Dr. Yosio Kobayasi, preserved in the National Museum of Nature and Science. Part 2: Cordyceps (Elaphocordyceps) on Elaphomyces

Four holotype specimens of Cordyceps parasitic on Elaphomyces that were originally described by Dr. Yosio Kobayasi and Mr. Daisuke Shimizu were relocated, and TNS (National Museum of Nature and Science, Japan) registration numbers (TNS-F-number) were assigned: viz. Cordyceps minazukiensis (TNS-F-197989), C. ophioglossoides var. cuboides (TNS-F-230312), C. valvatistipitata (TNS-F-230284), and Cordyceps × jezoensoides (TNS-F-230286). Two specimens were selected as lectotypes: C. delicatistipitata (TNS-F-230293) and C. ophioglossoides f. alba (TNS-F-18223). Those taxa, except for Cordyceps × jezoensoides, have been treated as members of the genus Elaphocordyceps in the latest taxonomy. The new combination Elaphocordyceps × jezoënsoides (Kobayasi) is proposed. In addition, Elaphomyces asahimontanus Kobayasi, the host of Cordyceps delicatistipitata, is lectotypified (TNS-F-230293).     

The phylogenetic placement of hypocrealean insect pathogens in the genus Polycephalomyces: An application of One Fungus One Name

Understanding the systematics and evolution of clavicipitoid fungi has been greatly aided by the application of molecular phylogenetics. They are now classified in three families, largely driven by reevaluation of the morphologically and ecologically diverse genus Cordyceps. Although reevaluation of morphological features of both sexual and asexual states were often found to reflect the structure of phylogenies based on molecular data, many species remain of uncertain placement due to a lack of reliable data or conflicting morphological characters. A rigid, darkly pigmented stipe and the production of a Hirsutella-like anamorph in culture were taken as evidence for the transfer of the species Cordyceps cuboidea, Cordyceps prolifica, and Cordyceps ryogamiensis to the genus Ophiocordyceps. Data from ribosomal DNA supported these species as a single group, but were unable to infer deeper relationships in Hypocreales. Here, molecular data for ribosomal and protein coding DNA from specimens of Ophiocordyceps cuboidea, Ophiocordyceps ryogamiensis, Ophiocordyceps paracuboidea, Ophiocordyceps prolifica, Cordyceps ramosopulvinata, Cordyceps nipponica, and isolates of Polycephalomyces were combined with a broadly sampled dataset of Hypocreales. Phylogenetic analyses of these data revealed that these species represent a clade distinct from the other clavicipitoid genera. Applying the recently adopted single system of nomenclature, new taxonomic combinations are proposed for these species in the genus Polycephalomyces, which has been historically reserved for asexual or anamorphic taxa.     

Reassessment of type specimens of Cordyceps and its allies described by Dr. Yosio Kobayasi preserved in the mycological herbarium of the National Museum of Nature and Science (TNS). Part 1: the genus Torrubiella

Nineteen holotype specimens of the genus Torrubiella described by Dr. Yosio Kobayasi and Mr. Daisuke Shimizu were relocated and accession numbers (TNS-F number) were given. A new scientific name, Torrubiella plana Hiroki Sato, Ban, Masuya & Hosoya nom. nov. (TNS-F-12061), was proposed for T. minutissima Kobaysi & Shimizu (homonym of T. minutissima Lagarde). The other 18 species follow: T. alboglobosa (TNS-F-12067), T. aurantia (TNS-F-12069), T. corniformis (TNS-F-12064), T. ellipsoidea (TNS-F-12055), T. formosana (TNS-F-12059), T. fusiformis (TNS-F-234548), T. globosostipitata (TNS-F-12057), T. longissima (TNS-F-12071), T. mammillata (TNS-F-12060), T. miyagiana (TNS-F-12062), T. neofusiformis (TNS-F-12058), T. oblonga (TNS-F-12070), T. ooaniensis (TNS-F-12063), T. pallida (TNS-F-12789), T. rosea (TNS-F-12065), T. ryogamimontana (TNS-F-12058), T. ryukyuensis (TNS-F-11932), and T. superficialis (TNS-F-12072).     

Molecular evidence of a teleomorph-anamorph connection between Cordyceps scarabaeicola and Beauveria sungii and its implication for the systematics of Cordyceps sensu stricto

A teleomorph of Beauveria sungii S.A. Rehner & Humber is identified as Cordyceps scarabaeicola Y. Kobayasi based on the teleomorphic material of the B. sungii isolate used in the recent phylogenetic study. Cordyceps Fr. 1818 is an older generic name than Beauveria Vuill. 1912 in Cordyceps sensu stricto. Here, C. scarabaeicola is suggested to be adopted for B. sungii on the priority basis of both generic name and species epithet in compliance with the recent revision of Article 59 of the Melbourne Code (recently changed to the ‘International Code of Nomenclature for algae, fungi, and plant’ or ICN). The pros and cons of a unified nomenclature for pleomorphic fungi are briefly discussed with reference to Beauveria.     

New teleomorph combinations in the entomopathogenic genus Metacordyceps

The genus Metacordyceps contains arthropod pathogens in Clavicipitaceae (Hypocreales) that formerly were classified in Cordyceps sensu Kobayasi et Mains. Of the current arthropod pathogenic genera of Hypocreales, the genus Metacordyceps remains one of the most poorly understood and contains a number of teleomorphic morphologies convergent with species of Cordyceps s.s. (Cordycipitaceae) and Ophiocordyceps (Ophiocordycipitaceae). Of note, the anamorph genera Metarhizium and Pochonia were found to be associated only with Metacordyceps and demonstrated to be phylogenetically informative for the clade. Several species of Cordyceps considered to have uncertain placements (incertae sedis) in the current taxonomic framework of clavicipitoid fungi were collected during field expeditions mostly in eastern Asia. Species reclassified here in Metacordyceps include Cordyceps atrovirens Kobayasi & Shimizu, Cordyceps indigotica Kobayasi & Shimizu, Cordyceps khaoyaiensis Hywel-Jones, Cordyceps kusanagiensis Kobayasi & Shimizu, Cordyceps martialis Speg., Ophiocordyceps owariensis Kobayasi, Cordyceps pseudoatrovirens Kobayasi & Shimizu and Ophicordyceps owariensis f. viridescens (Uchiy. & Udagawa) G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora. Incorporation of these species in a multigene phylogenetic framework of the major clades of clavicipitoid fungi more than doubled the number of species in Metacordyceps and allowed for refinement of morphological concepts for the genus consistent with the phylogenetic structure. Based on these findings we then discuss evolution of this genus, subgeneric relationships, anamorph connections, and suggest additional species that should be confirmed for possible inclusion in Metacordyceps.     

Teleomorph-anamorph relationships and reclassification of <Emphasis Type="Italic">Cordyceps cuboidea</Emphasis> and its allied species

Based on morphological characteristics and molecular phylogeny, we reclassified Cordyceps cuboidea and allied species C. alboperitheciata, C. prolifica, and Ophiocordyceps ryogamiensis. We investigated their teleomorph-anamorph relationships and revealed that these four species have Hirsutella-like anamorphs with morphological differences between them. By analyzing their molecular phylogeny, inferred from DNA sequences of internal transcribed spacer (ITS) and large subunit (LSU) D1/D2 region of rDNA, they were separated into four close-knit clades. Although C. prolifica and O. ryogamiensis formed their own clades, isolates of C. cuboidea separated into two clades, i.e., a true C. cuboidea clade and one resembling a new species, the O. paracuboidea clade. The latter two species are distinguished by the fruiting region of the stroma. In addition, C. alboperitheciata is regarded as a synonym of C. cuboidea. From the morphology, teleomorph-anamorph relationships, and molecular phylogeny, we concluded these species should be assigned to the genus Ophiocordyceps.     

高雄山虫草及其细脚拟青霉无性型

报道采自安徽省天堂寨自然保护区的高雄山虫草Cordyceps takaomontana及其无性型细脚拟青霉Paecilomyces tenuipes,应用单子囊孢子分离鉴定和微循环产孢的方法确证了两者的对应关系,并修订了高雄山虫草的原始描述。双梭孢虫草C．Bifusispora可能为高雄山虫草的同物异名。同时采集的具数个红色子座且有双梭形子囊孢子的虫草不是同一个种。     

Un nuovo ascomicete entomogeno rinvenuto in grotta: Cordyceps riverae

Abstract

A new entomogenous Ascomycete found in cave: Cordyceps riverae.—A new species of entomogenous Cordyceps, belonging to Subgenus Racemella (Ces.) Sacc., found on an adult Lepidotteron in a cave near L'Aquila (Abruzzo, Italy) is described. The characters of this species, named Cordyceps riverae, are compared with those of other similar species of Cordyceps.     

Cordyceps owariensis f. viridescens and its new Nomuraea anamorph

Cordyceps owariensis f. viridescens forma nov. (Nom. Jap., Amami-yarinohosemitake) is described from an underground nymph of Platypleura kuroiwae collected in Amami-oshima Island, southwestern Japan. Cultural isolations were made from ascospores of the fresh material of this fungus, and consequently a new hyphomycete was developed as its anamorph. Nomuraea owariensis is described to accommodate this anamorphic state. This is the first report on the association of a Nomuraea anamorph with cicadicolous Cordyceps species. Received: June 6, 2001 / Accepted: December 18, 2001     

The medicinal fungus Cordyceps militaris: research and development

Ophiocordyceps sinensis (syn. Cordyceps sinensis) is a highly valued medicinal fungus. This entomopathogen has a limited distribution, has been overharvested in the wild, and its stromata have not been artificially cultivated. Another entomopathogenic fungus, Cordyceps militaris (commonly known as orange caterpillar fungus), has chemical capacities similar to those of O. sinensis, but unlike O. sinensis, its stromata can be easily cultivated. Consequently, C. militaris is being studied as an alternative to O. sinensis, and the large-scale production of stromata is receiving substantial attention. Significant research has been conducted on the genetic resources, nutritional and environmental requirements, mating behavior, and biochemical and pharmacological properties of C. militaris. The complete genome of C. militaris has recently been sequenced. This fungus has been the subject of many reviews, but few have focused on its biology. The current paper reviews the biological aspects of the fungus including host range, mating system, cytology and genetics, insect- and non-insect nutritional requirements, environmental influence on stroma development, and commercial development.     

Genetics of Cordyceps and related fungi

Ascomycete Cordyceps sensu lato consists of hundreds of species of fungi capable of infecting different insects. Species of these fungi are either valued traditional Chinese medicines or used for biocontrol of insect pests. Phylogenomic analysis indicated that fungal entomopathogenicity has evolved for multiple times, and the species of Cordyceps were diverged from the mycoparasite or plant endophyte. Relative to plant pathogens and saprophytes, Cordyceps species demonstrate characteristic genome expansions of proteases and chitinases that are used by the fungi to target insect cuticles. Only a single mating-type gene identified in the sequenced species of Cordyceps sensu lato indicates that these fungi are sexually heterothallic, but the gene structure of the mating-type loci and frequency in performing sexual cycle are considerably different between different species. Similar to the model fungus Neurospora crassa, Cordyceps and related fungi contain the full components for RNA interference pathways. However, the mechanism of repeat-induced point mutation varies between different fungi. Epigenetic rather than genetic alterations are majorly responsible for the frequent occurrence of culture degeneration in Cordyceps-related species. Future genetic and epigenetic studies of fungal sexuality controls and culture degeneration mechanisms will benefit the cost-effective applications of Cordyceps and related fungi in pharmaceuticals and agriculture.     

Cordyceps cuncunae (Ascomycota, Hypocreales), a new pleoanamorphic species from temperate rainforest in southern Chile

Cordyceps cuncunae Palfner sp. nov. is reported from Valdivian rainforest in southern Chile, parasiting larvae of an unidentified ghost moth species (Lepidoptera, Hepialidae) which probably feed on roots of Laureliopsis philippiana. Morphology and anatomy of stromata as well as morphological and molecular characteristics of mycelium in pure culture which produces two anamorphs, one of them Lecanicillium-like, are described. The systematic position of the new taxon within the most recent generic concept is discussed. This is the first record of an endemic Cordyceps species from Chile.     

Heterothallism in Cordyceps takaomontana

Perithecium formation of an entomopathogenic fungus Cordyceps takaomontana was promoted by treating the mycelia with cell wall-degrading enzymes and PEG 4000. Perithecia were formed in the mixed culture of both mating-type strains MAT1 and MAT2, and not in the culture of MAT1 or MAT2 alone. The MAT1 strains did not possess a mating-type gene MAT1-1-3, but could produce perithecia. These results strongly suggested that C. takaomontana is heterothallic, and does not need MAT1-1-3 for the perithecium formation. MAT1-1-3 was also not found in another entomopathogenic fungus Cordyceps militaris. On the other hand, phytopathogenic fungi Balansia sp., Claviceps purpurea and Epichloë typhina possessed MAT1-1-3. The structures of mating-type locus MAT1-1 of these phytopathogenic fungi in the family Clavicipitaceae were similar to that of a phytopathogenic fungus Gibberella fujikuroi in the family Nectriaceae, which is closely related to Clavicipitaceae. These results suggested that phytopathogen might be more ancestral group than entomopathogen in Clavicipitaceae, and that MAT1-1-3 might be lost in the course of the host shift from plants to insects.     

Variations of SSU rDNA group I introns in different isolates of Cordyceps militaris and the loss of an intron during cross-mating

Cordyceps militaris, the type species of genus Cordyceps, is one of the most popular mushrooms and a nutraceutical in eastern Asia. It is considered a model organism for the study of Cordyceps species because it can complete its life cycle when cultured in vitro. In the present study, the occurrence and sequence variation of SSU rDNA group I introns, Cmi.S943 and Cmi.S1199, among different isolates of C. militaris were analyzed. Based on the secondary structure predictions, the Cmi.S943 intron has been placed in subgroup IC1, and the Cmi.S1199 intron has been placed in subgroup IE. No significant similarity between Cmi.S943 and Cmi.S1199 suggested different origins. Three genotypes, based on the frequency and distribution of introns, were described to discriminate the 57 surveyed C. militaris strains. It was found that the genotype was related to the stroma characteristics. The stromata of all of the genotype II strains, which possessed only Cmi.S943, could produce perithecium. In contrast, the stromata of all genotype III strains, which had both Cmi.S943 and Cmi.S1199, could not produce perithecium. Cmi.S1199 showed the lowest level of intra-specific variation among the tested strains. Group I introns can be lost during strain cross-mating. Therefore, we presumed that during cross-mating and recombination, intron loss could be driven by positive Darwinian selection due to the energetic cost of transcribing long introns.     

Cordylactam,a new alkaloid from the spider pathogenic fungus Cordyceps sp. BCC 12671

Cordylactam (1), a new lactam-fused 4-pyrone was isolated from the spider pathogenic fungus Cordyceps sp. BCC 12671. The structure was elucidated on the basis of NMR spectroscopic and mass spectrometry data, which was further confirmed by the spectroscopic analyses of a semisynthetic derivative. This is the first report of a new compound from spider pathogenic Cordyceps species.     

<Emphasis Type="Italic">Cordyceps bassiana</Emphasis> inhibits smooth muscle cell proliferation via the ERK1/2 MAPK signaling pathway

Cordyceps belongs to a genus of acormycete fungi and is known to exhibit various pharmacological effects. The aim of this study was to investigate the effect of Cordyceps species on the proliferation of vascular smooth muscle cells (VSMC) and their underlying molecular mechanism. A cell proliferation assay showed that Cordyceps bassiana ethanol extract (CBEE) significantly inhibited VSMC proliferation. In addition, neointimal formation was significantly reduced by treatment with CBEE in the carotid artery of balloon-injured rats. We also investigated the effects of CBEE on the extracellular signal-regulated kinase (ERK) signal pathway. Western blot analysis revealed increased ERK 1/2 phosphorylation in VSMCs treated with CBEE. Pretreatment with U0126 completely abrogated CBEE-induced ERK 1/2 phosphorylation. In conclusion, CBEE exhibited anti-proliferative properties that affected VSMCs through the ERK1/2 MAPK signaling pathway. Our data may elucidate the inhibitory mechanism of this natural product.     

Preservation affects the vegetative growth and fruiting body production of <Emphasis Type="Italic">Cordyceps militaris</Emphasis>

Cordyceps militaris is a model species of Cordyceps fungi, and has been traditionally used as an edible and medicinal fungus due to its richness of bioactive and pharmacological metabolites. The fruiting bodies of this fungus are widely used as healthy food and nutrition supply. In industrial production, fruiting bodies are cultivated on artificial media, but their yield and quality are usually affected by the quality of fungal strains. In this study, the effect of colony growth rate of the fungal strains, fungal age and repeated subculturing on the fungal biomass accumulation was investigated. The results indicated that the fungal biomass was positively correlated with the colony growth rate and not affected by fungal age and the repeated subculturing. The preservation conditions for stock cultures, including choice of cultures, lyophilization, temperature and protective agents were optimized based on the mycelial formation and conidia production in artificial inoculum. The development of fruiting bodies from the fungal strains stored under the optimized preservation conditions were further analyzed to determine the ideal time period of preservation. Results indicated that storing the fungus at 4 °C could maintain the fungal vitality and fruiting body producing capacity for at least 12 months. This study established practical criteria of fungal inoculum for artificial cultivation of fruiting body and provided a simple and efficient preservation method for C. militaris. The results may shed light on preservation for other Cordyceps species and other edible fungi.     

Bioactive naphthoquinones from Cordyceps unilateralis

Six bioactive naphthoquinone derivatives, erythrostominone, deoxyerythrostominone, 4-O-methyl erythrostominone, epierythrostominol, deoxyerythrostominol and 3,5,8-trihydroxy-6-methoxy-2-(5-oxohexa-1,3-dienyl)-1,4-naphthoquinone, were isolated from the insect pathogenic fungus Cordyceps unilateralis BCC1869. While the latter is synthetically known, both it and 4-O-methyl erythrostominone are products of fungus strain C. unilateralis BCC1869.