Two unusual new species of Pleosporales: Anteaglonium rubescens and Atrocalyx asturiensis.

Two new species of Pleosporales, Anteaglonium rubescens (Anteagloniaceae) and Atrocalyx asturiensis (Lophiotremataceae), are described. Phylogenetic placement was determined by combined analyses of a DNA data matrix containing ITS, LSU, SSU, rpb2, and tef1. Anteaglonium rubescens is a stromatic fungus characterized by brown didymospores disarticulating within asci, and by the production of a red-orange to pink pigment produced in nature and in artificial culture. Atrocalyx asturiensis has massive ascomatal crests and brown phragmospores.

Three former taxa of Cucurbitaria and considerations on Petrakia in the Melanommataceae.

Based on phylogenetic analyses of an ITS-LSU-SSU-rpb2-tef1 sequence data matrix three taxa once classified in Cucurbitaria are referred to the Melanommataceae. Cucurbitaria rhododendri, also known as Melanomma rhododendri, is not congeneric with the generic type of Melanomma, M. pulvis-pyrius and thus classified in the new genus Alpinaria. Cucurbitaria piceae, known as Gemmamyces piceae, the cause of the Gemmamyces bud blight of Picea spp., belongs also to the Melanommataceae. The name Gemmamyces is conserved. For Cucurbitaria obducens, also known as Teichospora obducens, the new genus Praetumpfia is described, as it cannot be accommodated in any known genus. All species are redescribed and epitypified. Based on sequence data and morphology, Blastostroma, Mycodidymella and Xenostigmina are synonyms of Petrakia. The genus Petrakia is emended. We also provide sequences of additional markers for Beverwykella pulmonaria, Melanomma pulvis-pyrius, Petrakia echinata and Pseudotrichia mutabilis.

Stigmatodiscus pruni, a new dothideomycete with hysteriform ascomata.

Stigmatodiscus pruni (Stigmatodiscaceae, Stigmatodiscales) is described and illustrated from corticated dead twigs of Prunus spinosa collected in Austria and France. It is characterised by hysteriform ascomata with two lateral black lips, which are erumpent through the periderm of the host, and a black disc in combination with two-celled, asymmetric brown verruculose ascospores with a distinct sheath. Phylogenetic analyses of a multigene matrix containing a representative selection of Dothideomycetes from four genes (nuc 18S rDNA, nuc 28S rDNA, rpb2 and tef1) revealed a highly supported placement within Stigmatodiscales as sister species to Stigmatodiscus enigmaticus. Micromorphology of the sexual and asexual morph matches the genus Stigmatodiscus, except for the hysteriform shape of the ascomata and the two-celled ascospores.

Asterodiscus and Stigmatodiscus, two new apothecial dothideomycete genera and the new order Stigmatodiscales.

During a survey on corticolous Dothideomycetes, several collections with ascospores matching the genera Asteromassaria and Stigmatomassaria (Pleomassariaceae, Pleosporales) were revealed from dead corticated twigs of Acer, Carpinus and Tamarix. Closer morphological examination showed that their ascomata were apothecial, with a hamathecium consisting of septate, branched paraphyses, which are apically swollen at maturity. Several collections were cultured and sequenced, and a Blast search of their nuc 28S rDNA sequences revealed dothideomycetous affiliation, but without a close match to a specific family or order. Phylogenetic analyses of a multigene matrix containing a representative selection of Dothideomycetes from four genes (nuc 18S rDNA, nuc 28S rDNA, rpb2 and tef1) revealed placement within Dothideomycetes but without a supported familial or ordinal affiliation. Based on the phylogenetic analyses and morphological investigations, the new genera Asterodiscus and Stigmatodiscus, with the two new species A. tamaricis and S. enigmaticus, are described and illustrated, and placed in the new family Stigmatodiscaceae and new order Stigmatodiscales.

Two new species of Thyronectria from Mediterranean Europe.

Two new species of Thyronectria growing in Mediterranean vegetation are described from southern Spain; they are T. giennensis from Quercus ilex ssp. rotundifolia and T. pistaciae from Pistacia lentiscus. Both species are characterized by morphology of sexual and asexual morphs and by DNA data. They have olivaceous to green-brown muriform ascospores and are closely related to T. asturiensis and T. roseovirens, as determined by multigene phylogenetic analyses of a matrix containing six loci (ITS and 28S regions of nuc rDNA, ACT1, RPB1, RPB2, TEF1 and TUB2 genes). We also report that Cucurbitaria bicolor is a synonym of Thyronectria rhodochlora, the type species of Thyronectria.

Taxonomic position of the genus Bicornispora and the appearance of a new species Bicornispora seditiosa.

The discovery of a second species of Bicornispora in Spain, B. seditiosa, which is closely related to B. exophiala but has smaller ascospores, narrower asci and different ecology, gave us the opportunity to culture and sequence the fungus. Phylogenetic analyses of rDNA regions including partial nuc 28S rDNA (28S) and ITS1-5.8S-ITS2 (ITS) supported a close relationship with species of the genus Lambertella (Rutstroemiaceae), placing Bicornispora, previously ascribed to Coryneliales, within Helotiales. This result confirmed an evolutionary linkage between certain inoperculate discomycetes such as Lambertella palmeri and derived cleistothecial forms (Bicornispora spp.). Based on analyses of morphological study and molecular phylogenetic analyses, a new combination Rutstroemia asphodeli is proposed for Ciboria asphodeli.

The rise and fall of Sarawakus (Hypocreaceae, Ascomycota).

Species of Sarawakus are rarely encountered. Their teleomorphs resemble sexual stages of Trichoderma, formerly called Hypocrea, but differ from that genus by unicellular ascospores. The two green-spored species S. britannicus and the type species of Sarawakus, S. lycogaloides, recently were collected, compared with their types and cultured. We redescribe and illustrate these species and transfer them to Trichoderma, based on phylogenetic analysis of the translation elongation factor 1-alpha encoding gene (tef1), containing the two last introns and exon, and a part of the rpb2 gene, encoding the second largest RNA polymerase subunit. Trichoderma lycogaloides, was found to cluster with Hypocrea sulawesensis, an unusual species of Trichoderma, while T. britannicum is closely related to T. aerugineum of the Spinulosa clade. The anamorphs of the two examined species are characterized by (odd) verticillium-like conidiophores, large cylindrical phialides and conidia, which belong to the largest of those species forming green conidia, oval to subglobose in T. lycogaloides and oblong in T. britannicum. All species currently recognized in Sarawakus are transferred to Trichoderma, introducing the new combinations T. fragile, T. hexasporum, T. izawae, T. sordidum, T. subtrachycarpum, T. succisum and T. trachycarpum and the new name T. rosellum. Trichoderma trachycarpum is redescribed and illustrated from an isotype.

Front line defenders of the ecological niche! Screening the structural diversity of peptaibiotics from saprotrophic and fungicolous Trichoderma/Hypocrea species.

Approximately 950 individual sequences of non-ribosomally biosynthesised peptides are produced by the genus Trichoderma/Hypocrea that belong to a perpetually growing class of mostly linear antibiotic oligopeptides, which are rich in the non-proteinogenic α-aminoisobutyric acid (Aib). Thus, they are comprehensively named peptaibiotics. Notably, peptaibiotics represent ca. 80 % of the total inventory of secondary metabolites currently known from Trichoderma/Hypocrea. Their unique membrane-modifying bioactivity results from amphipathicity and helicity, thus making them ideal candidates in assisting both colonisation and defence of the natural habitats by their fungal producers. Despite this, reports on the in vivo-detection of peptaibiotics have scarcely been published in the past. In order to evaluate the significance of peptaibiotic production for a broader range of potential producers, we screened nine specimens belonging to seven hitherto uninvestigated fungicolous or saprotrophic Trichoderma/Hypocrea species by liquid chromatography coupled to electrospray high resolution mass spectrometry. Sequences of peptaibiotics found were independently confirmed by analysing the peptaibiome of pure agar cultures obtained by single-ascospore isolation from the specimens. Of the nine species examined, five were screened positive for peptaibiotics. A total of 78 peptaibiotics were sequenced, 56 (=72 %) of which are new. Notably, dihydroxyphenylalaninol and O-prenylated tyrosinol, two C-terminal residues, which have not been reported for peptaibiotics before, were found as well as new and recurrent sequences carrying the recently described tyrosinol residue at their C-terminus. The majority of peptaibiotics sequenced are 18- or 19-residue peptaibols. Structural homologies with 'classical representatives' of subfamily 1 (SF1)-peptaibiotics argue for the formation of transmembrane ion channels, which are prone to facilitate the producer capture and defence of its substratum.

Molecular systematics of Woswasia atropurpurea gen. et sp. nov. (Sordariomycetidae), a fungicolous ascomycete with globose ascospores and holoblastic conidiogenesis.

The monotypic genus Woswasia is described for wood-inhabiting and mycotrophic fungi classified in the Sordariomycetidae. It is characterized by unicellular, hyaline, globose to ellipsoid, verruculose ascospores; unitunicate long-stipitate asci with an apical annulus staining blue in aqueous cotton blue and perithecia with a long neck immersed in a stroma exhibiting a conspicuous pH-dependent color reaction. In vitro, it produces branched subhyaline to hyaline conidiophores with terminally arranged sympodial conidiogenous cells and holoblastic hyaline conidia. The remarkable morphological similarity of Woswasia to Amplistroma of the Amplistromataceae, although suggestive of a close relationship, was not confirmed by molecular data. Phylogenetic analysis based on two functional ribosomal genes (large and small subunits of the nuclear rDNA) and one protein-coding gene (second largest subunit of RNA polymerase II) supports the placement of Woswasia in the Sordariomycetidae incertae sedis. Woswasia atropurpurea, the type and only species of the genus, groups within a large heterogeneous clade containing other small or monotypic genera of wood-inhabiting saprobic fungi, which are distantly related and of which the majority lacks an ordinal or familial affiliation. Within the clade a relationship of Woswasia to the freshwater genus Cyanoannulus is suggested.

New combinations in Trichoderma (Hypocreaceae, Hypocreales).

Unitary nomenclature demands the use of a single name for pleomorphic fungi determined according to priority. For this reason combinations in Trichoderma are here provided for 46 species for which such a combination is lacking. Although many more such species are known, only those are included here that are dealt with in more recent papers and where some DNA data are available in GenBank, even if erroneous; for other species it is strongly recommended to consult databases like Index Fungorum or MycoBank. Information on types is provided for most species, and representative cultures, GenBank accessions for tef1 and rpb2, and important references are given for all species.

Hypocrea britdaniae and H. foliicola: two remarkable new European species.

Two new species of Hypocrea are added here to the European funga. Hypocrea britdaniae, a fungus with unknown anamorph and large, conspicuous stromata resembling basidiomata of a corticiaceous fungus, is a sister species to the Longibrachiatum clade, while H. foliicola, a leaf-dwelling species that forms pulvinate stromata, is recognized as an additional member of the pachybasium core group. Hypocrea foliicola sporulates in culture in a reduced verticillium-like manner, while it produces a white, typical pachybasium-like anamorph in nature. Ecologically H. foliicola is remarkable in inhabiting leaves, a substrate rarely recorded for Hypocrea. All relevant morphological teleomorphic and anamorphic traits are given. The phylogenetic placement of the new species within Hypocrea/Trichoderma was determined with combined analyses of rpb2 and tef1 exon sequences.

Blue pigment in Hypocrea caerulescens sp. nov. and two additional new species in sect. Trichoderma.

Three new species of Hypocrea/Trichoderma sect. Trichoderma (Hypocreaceae, Hypocreales, Ascomycota, Fungi) are described from recent collections in southern Europe and the Canary Islands. They have been characterized by morphological and molecular methods, including microscopic examination of the teleomorph in thin sections, the anamorph, growth rate experiments and phylogenetic analyses based on a part of the translation elongation factor 1-alpha encoding gene (tef1) containing the two last introns and a part of the rpb2 gene, encoding the second largest RNA polymerase subunit. Analyses involving tef1 did not unequivocally resolve the sister clade relationship of Hypocrea caerulescens relative to the Koningii and Viride clades, while analyses based on rpb2 clearly suggest a close relationship with the former, although the phenotype of H. caerulescens is similar to H. viridescens, particularly by its warted conidia and a coconut-like odor in CMD culture. Hypocrea hispanica and T. samuelsii however are clearly related to the Viride clade by both phylogenetic markers, despite their morphological similarity to H. koningii and its relatives. An apparently specific blue pigment is formed in CMD cultures by Hypocrea caerulescens but could not be obtained by extraction with organic solvents.

New and interesting penzigioid Xylaria species with small, soft stromata.

Six penzigioid Xylaria species that are characterized by small, discoid to pulvinate, soft stromata are included in this study. Xylaria albocinctoides, X. bicampaniformis and X. lechatii are described as new; Nummularia albocincta, Hypoxylon carabayense and H. discolor are moved to the genus Xylaria to form new combinations X. albocincta, X. carabayensis and X. discolor respectively. An identification key is provided for the major aggregates of Xylaria that harbor penzigioid species as well as the species of the X. frustulosa aggregate, to which the six studied penzigioid Xylaria species belong.

The Longibrachiatum Clade of Trichoderma: a revision with new species.

The Longibrachiatum Clade of Trichoderma is revised. Eight new species are described (T. aethiopicum, T. capillare, T. flagellatum, T. gillesii, T. gracile, T. pinnatum, T. saturnisporopsis, T. solani). The twenty-one species known to belong to the Longibrachiatum Clade are included in a synoptic key. Trichoderma parareesei and T. effusum are redescribed based on new collections or additional observations. Hypocrea teleomorphs are reported for T. gillesii and T. pinnatum. Previously described species are annotated.

Nectria eustromatica sp. nov., an exceptional species with a hypocreaceous stroma.

A new species with remarkable morphology, Nectria eustromatica, is described, based on morphology of the teleomorph and anamorph, ecology and molecular phylogenetic analyses. Nectria eustromatica is characterized by sphaeroid perithecia immersed in pseudoparenchymatous stromata formed singly or collectively on a subiculum. Despite its deviating teleomorph morphology, it is placed within Nectria sensu stricto in phylogenetic analyses of a combined dataset of LSU, ITS, rpb2 and tef1 sequences with high internal support. Nectria eustromatica has been collected specifically on Hippocrepis (Coronilla) emerus in southern Europe. The anamorph of N. eustromatica shares morphological traits with the genera Stilbella and Tubercularia but produces non-phialidic macroconidia in addition to phialoconidia.

Stromatonectria gen. nov. and notes on Myrmaeciella.

Myrmaeciella caraganae was recollected in and around Vienna, Austria and found to be morphologically different generically from the type species of Myrmaeciella, M. endoleuca. We redescribe M. caraganae in the new genus Stromatonectria. Phylogenetic analyses of LSU sequences place the genus in the Bionectriaceae, Hypocreales. S. caraganae occurs on branches of Caragana spp., Colutea arborescens and Laburnum anagyroides of the Fabaceae. It is characterized by spheroid perithecia partly or entirely immersed in a Hypocrea-like stroma, a Nectria-like centrum and bicellular hyaline ascospores. Conidia of S. caraganae are produced in compound pycnidia that are formed prior to or in association with perithecia. Sporodochia but no pycnidia are formed in culture. We discuss the genus Myrmaeciella and compare S. caraganae with species of the Nectriaceae, including Nectria balansae, N. eustromatica and N. paraguayensis.

European species of Hypocrea part II: species with hyaline ascospores.

To date 75 species of Hypocrea/Trichoderma forming teleomorphs are recognised in Europe. The 56 hyaline-spored species are here described in detail and illustrated in colour plates, including cultures and anamorphs. This number includes 16 new holomorphs, two new teleomorphs and nine anamorphs of species previously described as teleomorphs. Phylogenetic placement and relationships of the species are shown on the strict consensus tree, based on sequences of RNA polymerase II subunit b (rpb2) and translation elongation factor 1 alpha (tef1) exon, comprising 135 species of the genus Hypocrea/Trichoderma. All available holotypes of species described from Europe including some from North America have been examined. A dichotomous key to the species is provided primarily utilising ecological and morphological traits of the teleomorphs and, where necessary, morphology of the anamorphs and cultures, and growth rates. Species descriptions are subdivided among five chapters, arranged primarily according to the larger phylogenetic clades, viz. section Trichoderma with 13 species, the pachybasium core group with 13 species including four species with stipitate stromata ('Podostroma'), species forming large effused stromata with 10 species including the section Hypocreanum, 9 species of the Brevicompactum, Lutea and Psychrophila clades, and 11 residual species of various smaller clades or of unknown phylogenetic placement. Finally, a list comprising dubious names and species excluded from Hypocrea that are relevant for Europe, or species claimed to occur in Europe by other authors is provided. Hypocrea minutispora is by far the most common species in Europe. For H. moravica, H. subalpina and H. tremelloides the anamorphs are newly described. The anamorphs of the latter two species and H. sambuci produce hyaline conidia on unusual structures new to Trichoderma. These three species form a new subclade of the morphologically strikingly different section Longibrachiatum, which is currently only represented by H. schweinitzii in Europe as a holomorph. The subclade is not named yet formally due to low statistical support. H. fungicola f. raduli is described as the new species H. austriaca, while H. hypomycella was found not to belong to Hypocrea. The typification of H. pilulifera, H. tremelloides and H. lutea has been clarified. Gliocladium deliquescens, the anamorph of H. lutea, is combined in Trichoderma. Species are epitypified where appropriate. Anamorph names are established prospectively to avoid numerous new combinations in future when they may be possibly used as holomorphic names if the ICBN is altered accordingly.

Distribution, Function, and Evolution of a Gene Essential for Trichothecene Toxin Biosynthesis in Trichoderma.

Trichothecenes are terpenoid toxins produced by species in 10 fungal genera, including species of Trichoderma. The trichothecene biosynthetic gene (tri) cluster typically includes the tri5 gene, which encodes a terpene synthase that catalyzes formation of trichodiene, the parent compound of all trichothecenes. The two Trichoderma species, Trichoderma arundinaceum and T. brevicompactum, that have been examined are unique in that tri5 is located outside the tri cluster in a genomic region that does not include other known tri genes. In the current study, analysis of 35 species representing a wide range of the phylogenetic diversity of Trichoderma revealed that 22 species had tri5, but only 13 species had both tri5 and the tri cluster. tri5 was not located in the cluster in any species. Using complementation analysis of a T. arundinaceum tri5 deletion mutant, we demonstrated that some tri5 homologs from species that lack a tri cluster are functional, but others are not. Phylogenetic analyses suggest that Trichoderma tri5 was under positive selection following its divergence from homologs in other fungi but before Trichoderma species began diverging from one another. We propose two models to explain these diverse observations. One model proposes that the location of tri5 outside the tri cluster resulted from loss of tri5 from the cluster in an ancestral species followed by reacquisition via horizontal transfer. The other model proposes that in species that have a functional tri5 but lack the tri cluster, trichodiene production provides a competitive advantage.

Clonal species Trichoderma parareesei sp. nov. likely resembles the ancestor of the cellulase producer Hypocrea jecorina/T. reesei.

We have previously reported that the prominent industrial enzyme producer Trichoderma reesei (teleomorph Hypocrea jecorina; Hypocreales, Ascomycota, Dikarya) has a genetically isolated, sympatric sister species devoid of sexual reproduction and which is constituted by the majority of anamorphic strains previously attributed to H. jecorina/T. reesei. In this paper we present the formal taxonomic description of this new species, T. parareesei, complemented by multivariate phenotype profiling and molecular evolutionary examination. A phylogenetic analysis of relatively conserved loci, such as coding fragments of the RNA polymerase B subunit II (rpb2) and GH18 chitinase (chi18-5), showed that T. parareesei is genetically invariable and likely resembles the ancestor which gave raise to H. jecorina. This and the fact that at least one mating type gene of T. parareesei has previously been found to be essentially altered compared to the sequence of H. jecorina/T. reesei indicate that divergence probably occurred due to the impaired functionality of the mating system in the hypothetical ancestor of both species. In contrast, we show that the sexually reproducing and correspondingly more polymorphic H. jecorina/T. reesei is essentially evolutionarily derived. Phenotype microarray analyses performed at seven temperature regimens support our previous speculations that T. parareesei possesses a relatively high opportunistic potential, which probably ensured the survival of this species in ancient and sustainable environment such as tropical forests.

The genus Melanconis (Diaporthales).

The genus Melanconis (Melanconidaceae, Diaporthales) in the strict sense is here re-evaluated regarding phylogenetic structure, taxonomy, distribution and ecology. Using a matrix of sequences from ITS, LSU, ms204, rpb2, tef1 and tub2, eight species are recognised and their phylogenetic positions are determined. Based on phylogenetic, morphological and geographical differentiation, Melanconis marginalis is subdivided into four subspecies. Melanconis italica is reduced to a subspecies of Melanconis marginalis. The two species Melanconis larissae from Betula sp. and M. pacifica from Alnus rubra are described as new. Melanconis alni and M. stilbostoma are lectotypified and M. alni, M. marginalis and M. stilbostoma are epitypified. All GenBank sequences deposited as Melanconis alni are shown to actually represent M. marginalis and those as M. marginalis belong to the newly described M. pacifica. Currently, Alnus and Betula are the sole host genera of Melanconis. All species and subspecies are (re-)described and illustrated. In addition, the neotypification of Melanconium pterocaryae is here validated.