伴 さやか

Cordyceps is a genus of ascomycete fungi with some of them being edible and/or having a long tradition in Chinese medicine. The chemical characterization of a solvent extract of the entomopathogenic fungus Cordyceps bifusispora afforded four undescribed coumarins, bifusicoumarin A-D (1-4), along with previously reported metabolites (5-8). Structural elucidation was performed via NMR, UV and HRMS analyses, X-ray single crystal diffraction and experimental ECD. A high throughput resazurin reduction assay, that measures cell viability, indicated that 5 has a IC50 between 1 and 15 μM for several assayed tumor lines. Moreover, a protein-interaction network indicated that C. bifusispora is a promising source of additional antitumor metabolites based on SwissTargetPrediction software predictions.

Dr. Kobayasi and Mr. Shimizu described 31 species of Cordyceps infecting Lepidoptera. Holotype specimens of 14 species and two au-thentic specimens of one of the 31 species were rediscovered from a herbarium of the National Museum of Nature and Science (TNS). Registration numbers (TNS-F-number) were given to these 16 specimens, and one was lectotypified as follows. Holotypes: Metarhizium indigoticum TNS-F-230337; Yosiokobayasia kusanagiensis TNS-F-197994 (Clavicipitaceae); Beauveria hepialidicola (Kobayasi & Shimizu) Hirok. Sato, S. Ban & Hosoya, comb. nov. TNS-F-197986; Cordyceps ampullacea TNS-F-197981, Cordyceps militaris f. alba TNS-F-230340, Cordyceps ochraceostromata TNS-F-195471, and Cordyceps rosea TNS-F-197972 (Cordyceps sensu stricto, Cordycipitaceae); Ophiocordy-ceps aurantia TNS-F-195485, Ophiocordyceps cochlidiicola TNS-F-195470, and Ophiocordyceps hiugensis TNS-F-197978 (Ophiocordy-cipitaceae); and Cordyceps changpaishanensis TNS-F-195501, Cordyceps ootakiensis TNS-F-197976, Cordyceps shimizui TNS-F-197995, and Cordyceps sulfurea TNS-F-197974 (Cordyceps sensu lato). Lectotype: Cordyceps bulolensis TNS-F-230327 (Cordyceps sensu lato). A new combination Beauveria hepialidicola comb. nov., is proposed for Cordyceps hepialidicola based on morphological observations.

The Aspergillus series Nigri contains biotechnologically and medically important species. They can produce hazardous mycotoxins, which is relevant due to the frequent occurrence of these species on foodstuffs and in the indoor environment. The taxonomy of the series has undergone numerous rearrangements, and currently, there are 14 species accepted in the series, most of which are considered cryptic. Species-level identifications are, however, problematic or impossible for many isolates even when using DNA sequencing or MALDI-TOF mass spectrometry, indicating a possible problem in the definition of species limits or the presence of undescribed species diversity. To re-examine the species boundaries, we collected DNA sequences from three phylogenetic markers (benA, CaM and RPB2) for 276 strains from series Nigri and generated 18 new whole-genome sequences. With the three- gene dataset, we employed phylogenetic methods based on the multispecies coalescence model, including four single-locus methods (GMYC, bGMYC, PTP and bPTP) and one multilocus method (STACEY). From a total of 15 methods and their various settings, 11 supported the recognition of only three species corresponding to the three main phylogenetic lineages: A. niger, A. tubingensis and A. brasiliensis. Similarly, recognition of these three species was supported by the GCPSR approach (Genealogical Concordance Phylogenetic Species Recognition) and analysis in DELINEATE software. We also showed that the phylogeny based on benA, CaM and RPB2 is suboptimal and displays significant differences from a phylogeny constructed using 5 752 single-copy orthologous proteins; therefore, the results of the delimitation methods may be subject to a higher than usual level of uncertainty. To overcome this, we randomly selected 200 genes from these genomes and performed ten independent STACEY analyses, each with 20 genes. All analyses supported the recognition of only one species in the A. niger and A. brasiliensis lineages, while one to four species were inconsistently delimited in the A. tubingensis lineage. After considering all of these results and their practical implications, we propose that the revised series Nigri includes six species: A. brasiliensis, A. eucalypticola, A. luchuensis (syn . A. piperis), A. niger (syn. A. vinaceus and A. welwitschiae), A. tubingensis (syn. A. chiangmaiensis, A. costaricensis, A. neoniger and A. pseudopiperis) and A. vadensis. We also showed that the intraspecific genetic variability in the redefined A. niger and A. tubingensis does not deviate from that commonly found in other aspergilli. We supplemented the study with a list of accepted species, synonyms and unresolved names, some of which may threaten the stability of the current taxonomy.

A new antifungal polyketide, named hakuhybotric acid (1), was isolated from a cultured broth of a mycoparasitic fungus Hypomyces pseudocorticiicola FKI-9008, together with two known analogs, F2928-1 (2) and Cladobotric acid E (3). Their structures were elucidated by MS and NMR analyses. The structure of hakuhybotric acid was the two epoxy groups of F2928-1 converted to olefins. All compounds showed antifungal activity against four different species of Aspergillus spp., the causative agents of aspergillosis. It was suggested that mycoparasitic fungi are a useful source to search antifungal drug lead compounds.

In this study, we aimed to clarify the phylogenetic distribution of Exophiala dermatitidis in Japan and describe the characteristics of genotypes. We examined 67 clinical and environmental isolates that were morphologically identified and preserved as E. dermatitidis and we confirmed the identification on the basis of the ribosomal DNA internal transcribed spacer region. Genotype sequences were aligned and compared using maximum likelihood phylogenetic tree analyses of the ITS1 region. Additionally, the strains of each genotype were tested for mycological characteristics, such as growth temperature, growth rate, and drug sensitivity. The 67 strains examined were isolated from Japan, the United States, Brazil, Venezuela, and China. In accordance with the establishment of a phylogenetic tree for the ITS1 region, 45 of the 49 Japanese strains were classified as genotype A, two as genotype B, and two as genotype D (A2 according to the method of Matos et al. (2003)). Chinese strains were divided into genotypes A and D (A2), and South American strains were classified into genotypes A, B, B2, and C2, while all strains from the U.S. belonged to genotype A. New genotype groups B2 and C2 were identified in Brazilian and Venezuelan strains, respectively. There were no specific differences among the genotypes or isolated regions in the antifungal susceptibility test for all E. dermatitidis isolates. However, genotypes B2 and D (A2) exhibited growth at higher temperatures than the other genotypes.

A new compound, 3-O-(4-O-methyl-β-D-glucopyranosyl) xanthone (1) was isolated from the culture of Conoideocrella luteorostrata NBRC106950. The structure of 1 was mainly determined by 1H, 13C, 2D-NMR and HREIMS spectral analyses. The absolute configuration of 4-O-methylglucopyranosyl moiety was determined by the optical rotation of aqueous layer of hydrolyzed 1 as D-configuration.

During the search for new antitrypanosomal drug leads, four antitrypanosomal compounds, of three depsipeptides and one nortriterpenoid, were isolated from cultures of the mutant strain IU-3 of the insect pathogenic fungus Ophiocordyceps coccidiicola NBRC 100683. Their structures were identified by the analysis of high resolution-electron ionization (HR-EI)-MS and HR-FAB-MS, and H-1- and C-13-NMR spectra, including extensive two dimensional (2D)-heteronuclear NMR experiments, and comparison with literature data for destruxin A (1), destruxin B (2), destruxin E chlorohydrin (3) and helvolic acid (4). Compounds 1-4 showed in vitro antitrypanosomal activity against Trypanosome brucei brucei GUTat3.1 with IC50 values of 0.33, 0.16, 0.061 and 5.08 mu g/mL, respectively.

The entomogenous anamorphic fungus lsaria takamizusanensis has not been resolved clearly in its teleomorphic state. We succeeded in inducing ascostroma formation by incubating conidiomata of I takamizusanensis on cicada adults in a moist chamber. We observed the ascostroma and conducted a phylogenetic analysis based on ITS rDNA and EF-l alpha genes. The morphology of the ascostroma was identical to that of Cordyceps ryogamimontana. In the phylogenetic tree inferred from EF-la, the isolate from the partspores grouped with nine strains derived from conidia of I. takamizusanensis, which was distinct from a clade including Purpureocillium lilacinum. Moreover, a conidial structure identical to that of I. takamizusanensis was rediscovered on the holotype specimen of C. ryogamimontana. As a result, we propose a new name, Purpureocillium takamizusanense, which is a combination of the teleomorph-anamorph connection of C. ryogamimontana-I. takamizusanensis, in accordance with the 'one fungus, one name' concept of the International Code of Nomenclature for Algae, Fungi, and Plants (ICN).

Three new Ophiocordyceps species from Japan are described: 1) O. coenomyia, which is parasitic to awl-fly larvae; 2) O. arborescens; and 3) O. macroacicularis, which parasitize moth larvae. Ophiocordyceps coenomyia is characterized by a globose, alutaceous fertile part at the apex of cylindrical, luteofulvous-to-pale yellow stroma, immersed lanceolate perithecia, and ascospores dividing into partspores. Based on its morphology and phylogenetic position, the new species is closely related to O. heteropoda. The latter two species produce superficial oval perithecia at the apex of cylindrical stroma, multiseptate ascospores, and Hirsutella-type anamorphs on the culturemedia. However, they are distinguished by the shape and size of their stroma. Multi-locus sequence analysis revealed that O. arborescens and O. macroacicularis were in the same phylogenetic clade, which is a sister to the clade containing O. sinensis. The distribution of anamorphic types in Ophiocordyceps is discussed.

Opaliferin, a polyketide with a unique partial structure in which a cyclopentanone and tetrahydrofuran were connected with an external double bond, was isolated from the insect pathogenic fungus Cordyceps sp. NBRC 106954. The structure and relative configuration of opaliferin were determined by spectroscopic analysis and X-ray crystallography. The absolute configuration was established by anomalous dispersion effects in X-ray diffraction measurements on the crystal of di(p-bromobenzoyl) ester of opaliferin. A plausible biosynthetic pathway for opaliferin is proposed.

Ophiocordycipitaceae is a diverse family comprising ecologically, economically, medicinally, and culturally important fungi. The family was recognized due to the polyphyly of the genus Cordyceps and the broad diversity of the mostly arthropod-pathogenic lineages of Hypocreales. The other two cordyceps-like families, Cordycipitaceae and Clavicipitaceae, will be revised taxonomically elsewhere. Historically, many species were placed in Cordyceps, but other genera have been described in this family as well, including several based on anamorphic features. Currently there are 24 generic names in use across both asexual and sexual life stages for species of Ophiocordycipitaceae. To reflect changes in Art. 59 in the International Code of Nomenclature for algae, fungi, and plants (ICN), we propose to protect and to suppress names within Ophiocordycipitaceae, and to present taxonomic revisions in the genus Tolypocladium, based on rigorous and extensively sampled molecular phylogenetic analyses. When approaching this task, we considered the principles of priority, monophyly, minimizing taxonomic revisions, and the practical utility of these fungi within the wider biological research community.

During the search for new antitrypanosomal drug leads, three new antitrypanosomal compounds, cardinalisamides A-C (1-3), were isolated from cultures of the insect pathogenic fungus Cordyceps cardinalis NBRC 103832. Their structures were elucidated using MS analyses and extensive 2D-heteronuclear NMR. The absolute configurations of 1-3 were addressed by chemical degradation and Marfey's analysis. 1-3 showed in vitro antitrypanosomal activity against Trypanosoma brucei brucei with IC50 values of 8.56, 8.65 and 8.63 mu g ml(-1), respectively.

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. Published by Elsevier Ltd on behalf of The British Mycological Society.

Twenty taxa of Cordyceps s. l. (belonging to three genera in the latest taxonomic system) from Cicadidae had been described by Kobayasi and Shimizu. Among them, 13 holotype specimens of Cordyceps were rediscovered from preserved specimens partially without ordering, and their registration numbers (TNS-F-number) were given: viz. (1) Elaphocordyceps inegoensis TNS-F-230289, (2) Elaphocordyceps paradoxa TNS-F-230313, (3) Elaphocordyceps toriharamontana TNS-F-230288, (4) Ophiocordyceps heteropoda TNS-F-230294, (5) Ophiocordyceps longissima TNS-F-230285, (6) Ophiocordyceps prolifica f. terminalis TNS-F-230295, (7) Ophiocordyceps pseudolongissima TNS-F-197983, (8) Ophiocordyceps takaoensis TNS-F-3026, (9) Cordyceps kanzashiana TNS-F-198015, (10) Cordyceps kobayasii Koval' (a parts per thousand C. sinclairii Kobayasi, not C. sinclairii Berk.) TNS-F-212384, (11) Cordyceps minuta TNS-F-11933, (12) Cordyceps ramosopulvinata TNS-F-197979, and (13) Cordyceps ryogamimontana TNS-F-230292. Two specimens were selected as lectotype: viz. (14) Ophiocordyceps prolifica TNS-F-230300 and (15) Ophiocordyceps yakusimensis TNS-F-230287. A paratype specimen of (16) Cordyceps pleuricapitata TNS-F-197965, and an authentic specimen of (17) Cordyceps imagamiana TNS-F-197966, were also rediscovered.

A new Ingoldian hyphomycete was isolated from fallen leaves in Bach Ma National Park, Vietnam, and is described here as Condylospora vietnamensis. This fungus is different from four known Condylospora species in morphological characteristics, having U- or N-shaped conidia.

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 Ophiocardyceps (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 & Spatafbra. 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.

Adding a overlayer of liquid paraffin to formalin and subsequently removing it with oil-absorbent sheets in order to observe the specimen are very useful procedures when handling specimens preserved in formalin.

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 x 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 x jezoensoides, have been treated as members of the genus Elaphocordyceps in the latest taxonomy. The new combination Elaphocordyceps x jezoA << nsoides (Kobayasi) is proposed. In addition, Elaphomyces asahimontanus Kobayasi, the host of Cordyceps delicatistipitata, is lectotypified (TNS-F-230293).