About spirals and pores: Xylariaceae with remarkable germ loci.

Based on phylogenetic analyses of a multi-gene matrix of nuITS-LSU rDNA, RPB2 and TUB2 sequences and morphology, xylariaceous species with uni- to pauciperitheciate stromata and ascospores having a spirally coiling (helicoid) germ slit are revised and reclassified, including detailed descriptions and illustrations. The genus Helicogermslita is redefined and restricted to seven species with massive, erumpent, clypeus-like carbonaceous stromata, and Rosellinia somala is combined in Helicogermslita. Within the core Xylariaceae, the poorly known Leptomassaria simplex is shown to be closely related to Anthostoma insidiosum, for which the new genus Oligostoma is established, and Anthostoma rhenanum is demonstrated to be synonymous with O. insidiosum. The new genus Albicollum, characterised by immersed ascomata and a collar of white pseudostromatic tissues surrounding the ostioles, is established for Amphisphaeria canicollis, Anthostoma chionostomum, Sordaria (= Helicogermslita) fleischhakii and Anthostoma vincensii. Anthostoma ostropoides is synomymised with Albicollum canicolle, and Al. berberidicola, Al. longisporum and Al. novomexicanum are described as new species. Rosellinia (= Helicogermslita) gaudefroyi is transferred to the new genus Spiririma. Anthostoma amoenum and Euepixylon udum, both with a poroid germ locus, are shown to be only distantly related, and An. amoenum is reclassified within the asexual genus Digitodochium. Based on phylogeny, the genus Euepixylon is treated as a synonym of Nemania. A new species, Nemania ethancrensonii, which is closely related to the two formerly accepted Euepixylon species (E. sphaeriostomum, E. udum) but strongly deviates from the morphological concept of Euepixylon and Nemania, is described from the eastern USA. The genera Anthostomelloides, Clypeosphaeria, Digitodochium, Emarcaea, Induratia, Linosporopsis, Magnostiolata, Occultitheca and Spiririma are revealed to form a morphologically heterogeneous lineage in a basal position of Xylariaceae. Anthostoma vincensii, Quaternaria simplex and Rosellinia gaudefroyi are lectotypified, and Amphisphaeria canicollis, Anthostoma amoenum, An. rhenanum, An. vincensii, Quaternaria simplex, Rosellinia gaudefroyi and Valsa insidiosa are epitypified. Keys to uni- to pauciperitheciate xylariaceous genera with sigmoid to helicoid germ slits and to species of Albicollum are provided. Citation: Voglmayr H, Tello S, Jaklitsch WM, et al. 2022. About spirals and pores: Xylariaceae with remarkable germ loci. Persoonia 49: 58-98. https://doi.org/10.3767/persoonia.2022.49.02.

Multi-locus phylogenetic analysis of lophiostomatoid fungi motivates a broad concept of Lophiostoma and reveals nine new species.

Recent studies on the fungal families Lophiostomataceae and Lophiotremataceae (Pleosporales) have provided varying phylogenetic and taxonomic results concerning constituent genera and species. By adding DNA sequences of 24 new strains of Lophiostomataceae and nine new strains of Lophiotremataceae to a sequence data matrix from international databases, we provide a new understanding of the relationships within these families. Multigene analysis of the four molecular markers ITS, LSU, TEF1-α, and RPB2 reveals that the genera within Lophio-tremataceae are phylogenetically well supported. Lophiostoma myriocarpum is recognised as a species of Lophiotrema in contrast to earlier concepts. In Lophiostomataceae, we resurrect a broad generic concept of the genus Lophiostoma and reduce 14 genera to synonymy: Alpestrisphaeria, Biappendiculispora, Capulatispora, Coelodictyosporium, Guttulispora, Lophiohelichrysum, Lophiopoacea, Neopaucispora, Neotrematosphaeria, Platystomum, Pseudocapulatispora, Pseudolophiostoma, Pseudoplatystomum, and Sigarispora. Nine new species are described based on molecular data and in most cases supported by morphological characters: Antealophiotrema populicola, Atrocalyx nordicus, Lophiostoma carpini, Lophiostoma dictyosporium, Lophiostoma erumpens, Lophiostoma fusisporum, Lophiostoma jotunheimenense, Lophiostoma plantaginis, and Lophiostoma submuriforme. Lophiostoma caespitosum and Lophiotrema myriocarpum are lecto- and epitypified to stabilise their species concepts. High intraspecific variability of several morphological traits is common within Lophiostomataceae. Citation: Andreasen M, Skrede I, Jaklitsch WM, et al. 2021. Multi-locus phylogenetic analysis of lophiostomatoid fungi motivates a broad concept of Lophiostoma and reveals nine new species. Persoonia 46: 240-271. https://doi.org/10.3767/persoonia.2021.46.09.

Fenestelloid clades of the Cucurbitariaceae.

Fresh collections and their ascospore and conidial isolates backed up by type studies and molecular phylogenetic analyses of a multigene matrix of partial nuSSU-, complete ITS, partial LSU rDNA, rpb2, tef1 and tub2 sequences were used to evaluate the boundaries and species composition of Fenestella and related genera of the Cucurbitariaceae. Eight species, of which five are new, are recognised in Fenestella s.str., 13 in Parafenestella with eight new species and two in the new genus Synfenestella with one new species. Cucurbitaria crataegi is combined in Fenestella, C. sorbi in Synfenestella, Fenestella faberi and Thyridium salicis in Parafenestella. Cucurbitaria subcaespitosa is distinct from C. sorbi and combined in Neocucurbitaria. Fenestella minor is a synonym of Valsa tetratrupha, which is combined in Parafenestella. Cucurbitaria marchica is synonymous with Parafenestella salicis, Fenestella bavarica with S. sorbi, F. macrospora with F. media, and P. mackenziei is synonymous with P. faberi, and the latter is lectotypified. Cucurbitaria sorbi, C. subcaespitosa and Fenestella macrospora are lecto- and epitypified, Cucurbitaria crataegi, Fenestella media, F. minor and Valsa tetratrupha are epitypified in order to stabilise the names in their phylogenetic positions. A neotype is proposed for Thyridium salicis. A determinative key to species is given. Asexual morphs of fenestelloid fungi are phoma-like and do not differ from those of other representatives of the Cucurbitariaceae. The phylogenetic structure of the fenestelloid clades is complex and can only be resolved at the species level by protein-coding genes, such as rpb2, tef1 and tub2. All fungal species studied here occur, as far as has been possible to determine, on members of Diaporthales, most frequently on asexual and sexual morphs of Cytospora.

Two new classes of Ascomycota: Xylobotryomycetes and Candelariomycetes.

Phylogenetic analyses of a combined DNA data matrix containing nuclear small and large subunits (nSSU, nLSU) and mitochondrial small subunit (mtSSU) ribosomal RNA and the largest and second largest subunits of the RNA polymerase II (rpb1, rpb2) of representative Pezizomycotina revealed that the enigmatic genera Xylobotryum and Cirrosporium form an isolated, highly supported phylogenetic lineage within Leotiomyceta. Acknowledging their morphological and phylogenetic distinctness, we describe the new class Xylobotryomycetes, containing the new order Xylobotryales with the two new families Xylobotryaceae and Cirrosporiaceae. The two currently accepted species of Xylobotryum, X. andinum and X. portentosum, are described and illustrated by light and scanning electron microscopy. The generic type species X. andinum is epitypified with a recent collection for which a culture and sequence data are available. Acknowledging the phylogenetic distinctness of Candelariomycetidae from Lecanoromycetes revealed in previous and the current phylogenetic analyses, the new class Candelariomycetes is proposed.

Lichens or endophytes? The enigmatic genus Leptosillia in the Leptosilliaceae fam. nov. (Xylariales), and Furfurella gen. nov. (Delonicicolaceae).

Based on DNA sequence data, the genus Leptosillia is shown to belong to the Xylariales. Molecular phylogenetic analyses of ITS-LSU rDNA sequence data and of a combined matrix of SSU-ITS-LSU rDNA, rpb1, rpb2, tef1 and tub2 reveal that the genera Cresporhaphis and Liberomyces are congeneric with Leptosillia. Coelosphaeria fusariospora, Leptorhaphis acerina, Leptorhaphis quercus f. macrospora, Leptorhaphis pinicola, Leptorhaphis wienkampii, Liberomyces pistaciae, Sphaeria muelleri and Zignoëlla slaptonensis are combined in Leptosillia, and all of these taxa except for C. fusariospora, L. pinicola and L. pistaciae are epitypified. Coelosphaeria fusariospora and Cresporhaphis rhoina are lectotypified. Liberomyces macrosporus and L. saliciphilus, which were isolated as phloem and sapwood endophytes, are shown to be synonyms of Leptosillia macrospora and L. wienkampii, respectively. All species formerly placed in Cresporhaphis that are now transferred to Leptosillia are revealed to be non-lichenized. Based on morphology and ecology, Cresporhaphis chibaensis is synonymised with Rhaphidicyrtis trichosporella, and C. rhoina is considered to be unrelated to the genus Leptosillia, but its generic affinities cannot be resolved in lack of DNA sequence data. Phylogenetic analyses place Leptosillia as sister taxon to Delonicicolaceae, and based on morphological and ecological differences, the new family Leptosilliaceae is established. Furfurella, a new genus with the three new species, F. luteostiolata, F. nigrescens and F. stromatica, growing on dead branches of mediterranean fabaceous shrubs from tribe Genisteae, is revealed to be the closest relative of Delonicicola in the family Delonicicolaceae, which is emended. ITS rDNA sequence data retrieved from GenBank demonstrate that the Leptosilliaceae were frequently isolated or sequenced as endophytes from temperate to tropical regions, and show that the genus Leptosillia represents a widely distributed component of endophyte communities of woody plants.

A preliminary account of the Cucurbitariaceae.

Fresh collections, type studies and molecular phylogenetic analyses of a multigene matrix of partial nuSSU-ITS-LSU rDNA, rpb2, tef1 and tub2 sequences were used to evaluate the boundaries of Cucurbitaria in a strict sense and of several related genera of the Cucurbitariaceae. Two species are recognised in Cucurbitaria and 19 in Neocucurbitaria. The monotypic genera Astragalicola, Cucitella, Parafenestella, Protofenestella, and Seltsamia are described as new. Fenestella is here included as its generic type F. fenestrata (= F. princeps), which is lecto- and epitypified. Fenestella mackenzei and F. ostryae are combined in Parafenestella. Asexual morphs of Cucurbitariaceae, where known, are all pyrenochaeta- or phoma-like. Comparison of the phylogenetic analyses of the ITS-LSU and combined matrices demonstrate that at least rpb2 sequences should be added whenever possible to improve phylogenetic resolution of the tree backbone; in addition, the tef1 introns should be added as well to improve delimitation of closely related species.

Juglanconis gen. nov. on Juglandaceae, and the new family Juglanconidaceae (Diaporthales).

Molecular phylogenetic analyses of ITS-LSU rDNA sequence data demonstrate that Melanconis species occurring on Juglandaceae are phylogenetically distinct from Melanconis s.str., and therefore the new genus Juglanconis is described. Morphologically, the genus Juglanconis differs from Melanconis by light to dark brown conidia with irregular verrucae on the inner surface of the conidial wall, while in Melanconis s.str. they are smooth. Juglanconis forms a separate clade not affiliated with a described family of Diaporthales, and the family Juglanconidaceae is introduced to accommodate it. Data of macro- and microscopic morphology and phylogenetic multilocus analyses of partial nuSSU-ITS-LSU rDNA, cal, his, ms204, rpb1, rpb2, tef1 and tub2 sequences revealed four distinct species of Juglanconis. Comparison of the markers revealed that tef1 introns are the best performing markers for species delimitation, followed by cal, ms204 and tub2. The ITS, which is the primary barcoding locus for fungi, is amongst the poorest performing markers analysed, due to the comparatively low number of informative characters. Melanconium juglandinum (= Melanconis carthusiana), M. oblongum (= Melanconis juglandis) and M. pterocaryae are formally combined into Juglanconis, and J. appendiculata is described as a new species. Melanconium juglandinum and Melanconis carthusiana are neotypified and M. oblongum and Diaporthe juglandis are lectotypified. A short description and illustrations of the holotype of Melanconium ershadii from Pterocarya fraxinifolia are given, but based on morphology it is not considered to belong to Juglanconis. A key to all treated species of Juglanconis is provided.

Corynespora, Exosporium and Helminthosporium revisited - New species and generic reclassification.

Molecular phylogenetic analyses of a multigene matrix of partial nuSSU-ITS-LSU rDNA, rpb2 and tef1 sequences were performed to investigate the phylogenetic relationships of Corynespora, Exosporium and Helminthosporium species. Based on phylogenetic analyses and morphology, the genus Exosporium is synonymised with Helminthosporium, and the genus Corynespora is revealed as polyphyletic. Corynespora smithii is confirmed to be closely related to the generic type C. cassiicola and its morphology is described and illustrated. Exosporium tiliae, Corynespora caespitosa, C. endiandrae, C. leucadendri and C. olivacea are recognised in Helminthosporium, and Splanchnonema quercicola and S. kalakadense are combined in Helminthosporium. Based on pure culture studies and DNA sequence data, Massaria heterospora and Massarinula italica are shown to be the sexual morphs of Helminthosporium tiliae and H. microsorum, respectively. European accessions of Splanchnonema quercicola are recognised to differ from the North American type and are described as Helminthosporium quercinum. The sexual morph of H. oligosporum is recorded and described for the first time. The generic type of Helminthosporium, H. velutinum, is epitypified with a recent collection from the type host, Fagus sylvatica. Based on sequence data, Helminthosporium genistae is recognised as a distinct species. Several species for which subperidermal stromata have been reported are shown to be fungicolous on Diaporthales, the "stromata" representing aborted and transformed host stromata or conidiomata: H. caespitosum, H. microsorum, H. quercicola and H. quercinum on Coryneum spp.; H. hispanicum on conidiomata of Juglanconis juglandina; H. juglandinum on conidiomata of Diaporthe sp.; H. oligosporum and H. tiliae on Hercospora tiliae. The newly described H. austriacum is fungicolous on Amphisphaeria cf. millepunctata (Xylariales).

Hidden diversity in Thyridaria and a new circumscription of the Thyridariaceae.

A multigene analysis of a combined ITS-LSU-SSU-rpb2-tef1 sequence data matrix was applied to infer the phylogenetic position of the genus Thyridaria in the Pleosporales. The generic type of Thyridaria, T. broussonetiae (syn. T. incrustans), is situated in a clade currently named Roussoellaceae, which becomes a synonym of Thyridariaceae. However, Thyridaria rubronotata does not belong to this clade, but is here recognised as Cyclothyriella rubronotata in its own family Cyclothyriellaceae. The Thyridariaceae contain the genera Thyridaria, Roussoella, Roussoellopsis, Neoroussoella and the new genus Parathyridaria. Roussoella acaciae is combined in Thyridaria and Roussoella percutaenea in Parathyridaria. Ohleria modesta and an additional new thyridaria-like genus, Hobus, are found to represent isolated lineages with unresolved phylogenetic affinites within the Pleosporales. For Ohleria the new family Ohleriaceae is established. Melanomma fuscidulum belongs to Nigrograna, and three new species are described in this genus. A strain named Biatriospora marina clusters with Nigrograna. Based on the newly recognised species in Nigrograna, morphology and ecology do in no way correlate among these genera, therefore we erect the new family Nigrogranaceae for Nigrograna and recommend to discontinue the use of the family name Biatriosporaceae until fresh material of B. marina becomes available for sequencing.

Resolution of morphology-based taxonomic delusions: Acrocordiella, Basiseptospora, Blogiascospora, Clypeosphaeria, Hymenopleella, Lepteutypa, Pseudapiospora, Requienella, Seiridium and Strickeria.

Fresh material, type studies and molecular phylogeny were used to clarify phylogenetic relationships of the nine genera Acrocordiella, Blogiascospora, Clypeosphaeria, Hymenopleella, Lepteutypa, Pseudapiospora, Requienella, Seiridium and Strickeria. At first sight, some of these genera do not seem to have much in common, but all were found to belong to the Xylariales, based on their generic types. Thus, the most peculiar finding is the phylogenetic affinity of the genera Acrocordiella, Requienella and Strickeria, which had been classified in the Dothideomycetes or Eurotiomycetes, to the Xylariales. Acrocordiella and Requienella are closely related but distinct genera of the Requienellaceae. Although their ascospores are similar to those of Lepteutypa, phylogenetic analyses do not reveal a particularly close relationship. The generic type of Lepteutypa, L. fuckelii, belongs to the Amphisphaeriaceae. Lepteutypa sambuci is newly described. Hymenopleella is recognised as phylogenetically distinct from Lepteutypa, and Hymenopleella hippophaëicola is proposed as new name for its generic type, Sphaeria (= Lepteutypa) hippophaës. Clypeosphaeria uniseptata is combined in Lepteutypa. No asexual morphs have been detected in species of Lepteutypa. Pseudomassaria fallax, unrelated to the generic type, P. chondrospora, is transferred to the new genus Basiseptospora, the genus Pseudapiospora is revived for P. corni, and Pseudomassaria carolinensis is combined in Beltraniella (Beltraniaceae). The family Clypeosphaeriaceae is discontinued, because the generic type of Clypeosphaeria, C. mamillana, is a member of the Xylariaceae. The genus Seiridium, of which the sexual morph Blogiascospora is confirmed, is unrelated to Lepteutypa, as is Lepteutypa cupressi. The taxonomy of the cypress canker agents is discussed. The family Sporocadaceae is revived for a large clade of the Xylariales that contains Hymenopleella, Seiridium and Strickeria among a number of other genera. Neotypes are proposed for Massaria fuckelii and Sphaeria hippophaës. Didymella vexata, Seiridium marginatum, Sphaeria corni, Sphaeria hippophaës, Sphaeria seminuda are epitypified, Apiosporina fallax, Massaria occulta, Sphaeria mamillana and Strickeria kochii are lecto- and epitypified. We also provide DNA data for Broomella vitalbae, Cainia desmazieri and Creosphaeria sassafras.

Valsaria and the Valsariales.

More than 100 recent collections of Valsaria sensu lato mostly from Europe were used to elucidate the species composition within the genus. Multigene phylogeny based on SSU, LSU, ITS, rpb2 and tef1 sequences revealed a monophyletic group of ten species within the Dothideomycetes, belonging to three morphologically similar genera. This group could not be accommodated in any known family and are thus classified in the new family Valsariaceae and the new order Valsariales. The genus Valsaria sensu stricto comprises V. insitiva, V. robiniae, V. rudis, V. spartii, V. lopadostomoides sp. nov. and V. neotropica sp. nov., which are phylogenetically well-defined, but morphologically nearly indistinguishable species. The new monotypic genus Bambusaria is introduced to accommodate Valsaria bambusae. Munkovalsaria rubra and Valsaria fulvopruinata are combined in Myrmaecium, a genus traditionally treated as a synonym of Valsaria, which comprises three species, with M. rubricosum as its generic type. This work is presented as a basis for additional species to be detected in future.

Biodiversity of Trichoderma (Hypocreaceae) in Southern Europe and Macaronesia.

The first large-scale survey of sexual and asexual Trichoderma morphs collected from plant and fungal materials conducted in Southern Europe and Macaronesia including a few collections from French islands east of Africa yielded more than 650 specimens identified to the species level. Routine sequencing of tef1 revealed a genetic variation among these isolates that exceeds previous experience and ca. 90 species were recognized, of which 74 are named and 17 species newly described. Aphysiostroma stercorarium is combined in Trichoderma. For the first time a sexual morph is described for T. hamatum. The hitherto most complete phylogenetic tree is presented for the entire genus Trichoderma, based on rpb2 sequences. For the first time also a genus-wide phylogenetic tree based on acl1 sequences is shown. Detailed phylogenetic analyses using tef1 sequences are presented in four separate trees representing major clades of Trichoderma. Discussions involve species composition of clades and ecological and biogeographic considerations including distribution of species.

Phylogenetic and taxonomic revision of Lopadostoma.

The genus Lopadostoma (Xylariaceae, Xylariales) is revised. Most species formerly assigned to Lopadostoma do not belong to the genus. Twelve species are herein recognised, of which two are only known from morphology. Ten species, of which six (L. americanum, L. fagi, L. insulare, L. lechatii, L. meridionale and L. quercicola) are newly described, are characterised by both morphology and DNA phylogeny using LSU, ITS and rpb2 sequences. Morphologically, ecologically and phylogenetically Lopadostoma is a well-defined genus comprising exclusively species with pustular pseudostroma development in bark of angiospermous trees. Phaeosperma ailanthi, Phaeosperma dryophilum and Sphaeria linosperma are combined in Lopadostoma. Lopadostoma gastrinum is neotypified and L. turgidum is lecto- and epitypified. Species with asci and ascospores similar to those of Lopadostoma but having perithecia immersed in wood, particularly those of Lopadostoma subg. Anthostomopsis have been determined to be unrelated to the genus. DNA data confirm that Anthostoma is unrelated to Lopadostoma. Its type and currently only confirmed species Anthostoma decipiens belongs to Diatrypaceae. DNA data also show that L. pouzarii and Barrmaelia macrospora are unrelated to Lopadostoma. A commentary is provided for names in Lopadostoma and those names in Anthostoma that may be putative species of Lopadostoma based on their protologues. Anthostoma insidiosum is an older name for Anthostomella (Diatrype) adusta.

Stilbosporaceae resurrected: generic reclassification and speciation.

Following the abolishment of dual nomenclature, Stilbospora is recognised as having priority over Prosthecium. The type species of Stilbospora, S. macrosperma, is the correct name for P. ellipsosporum, the type species of Prosthecium. The closely related genus Stegonsporium is maintained as distinct from Stilbospora based on molecular phylogeny, morphology and host range. Stilbospora longicornuta and S. orientalis are described as new species from Carpinus betulus and C. orientalis, respectively. They differ from the closely related Stilbospora macrosperma, which also occurs on Carpinus, by longer, tapering gelatinous ascospore appendages and by distinct LSU, ITS rDNA, rpb2 and tef1 sequences. The asexual morphs of Stilbospora macrosperma, S.longicornuta and S. orientalis are morphologically indistinguishable; the connection to their sexual morphs is demonstrated by morphology and DNA sequences of single spore cultures derived from both ascospores and conidia. Both morphs of the three Stilbospora species on Carpinus are described and illustrated. Other species previously recognised in Prosthecium, specifically P.acerophilum, P. galeatum and P. opalus, are determined to belong to and are formally transferred to Stegonsporium. Isolates previously recognised as Stegonsporium pyriforme (syn. Prosthecium pyriforme) are determined to consist of three phylogenetically distinct lineages by rpb2 and tef1 sequence data, two of which are described as new species (S. protopyriforme, S. pseudopyriforme). Stegonsporium pyriforme is lectotypified and this species and Stilbospora macrosperma are epitypified. Based on DNA sequence data, the North American Stegonsporium acerophilum is recorded from Europe for the first time, and new hosts from Acer sect. Acer are reported for S. opalus and S. pyriforme. Stilbospora and Stegonsporium are classified within the revived family Stilbosporaceae. Prosthecium appendiculatum, P. auctum and P. innesii are shown to be unrelated to the Stilbosporaceae and are recognised in three distinct genera, Phaeodiaporthe appendiculata, Alnecium auctum n. gen. and Calosporella innesii within Diaporthaceae, Gnomoniaceae and Sydowiellaceae, respectively. The generic types of these three monotypic genera are briefly described, illustrated and lecto- and epitypfied.

Persistent hamathecial threads in the Nectriaceae, Hypocreales: Thyronectria revisited and re-instated.

Based on type studies and freshly collected material we here re-instate the genus Thyronectria (Nectriaceae, Hypocreales). Species of this genus were recently for the most part classified in the genera Pleonectria (Nectriaceae) or Mattirolia (Thyridiaceae), because Thyronectria and other genera had been identified as members of the Thyridiaceae due to the presence of paraphyses. Molecular phylogenies based on several markers (act, ITS, LSU rDNA, rpb1, rpb2, tef1, tub) revealed that the Nectriaceae contain members whose ascomata are characterised by long, more or less persistent, apical paraphyses. All of these belong to a single genus, Thyronectria, which thus has representatives with hyaline, rosy, green or even dark brown and sometimes distoseptate ascospores. The type species of Thyronectria, T. rhodochlora, syn. T. patavina, syn. T. pyrrhochlora is re-described and illustrated. Within the Nectriaceae persistent, apical paraphyses are common in Thyronectria and rarely also occur in Nectria. The genus Mattirolia is revised and merged with Thyronectria and also Thyronectroidea is regarded as a synonym of Thyronectria. The three new species T. asturiensis, T. caudata and T. obscura are added to the genus. Species recently described in Pleonectria as well as some species of Mattirolia are combined in the genus, and a key to Thyronectria is provided. Five species are epitypified. The type species of the genus Thyridium (Thyridiaceae), T. vestitum, is included in phylogenetic analyses to illustrate the phylogenetic distance of Thyronectria from the Thyridiaceae.

Disentangling the Trichoderma viridescens complex.

Trichoderma viridescens is recognised as a species complex. Multigene analyses based on the translation elongation factor 1-alpha encoding gene (tef1), a part of the rpb2 gene, encoding the second largest RNA polymerase subunit and the larger subunit of ATP citrate lyase (acl1) reveals 13 phylogenetic species with little or no phenotypic differentiation. This is the first use of acl1 in Trichoderma phylogenetics. The typification of T. viridescens s.str. is clarified and Hypocrea viridescens is replaced by the new name T. paraviridescens. Besides these two species, eleven are phylogenetically recognised and T. olivascens, T. viridarium, T. virilente, T. trixiae, T. viridialbum, T. appalachiense, T. neosinense, T. composticola, T. nothescens and T. sempervirentis are formally described and illustrated. Several species produce yellow diffusing pigment on cornmeal dextrose agar, particularly after storage at 15 °C, while T. olivascens is characterised by the formation of an olivaceous pigment. The results are compared with earlier publications on this group of species.

Intracellular location of enzymes involved in citrate production by Aspergillus niger.

The intracellular distribution and maximal activities of nine enzymes involved in the biosynthesis and degradation of citric acid in Aspergillus niger were determined under conditions of growth and of citric acid production. Under these conditions the intracellular location of the enzymes in most cases resembled that described for other filamentous fungi. Pyruvate carboxylase was found predominantly or exclusively in the cytosol. A single isoenzyme of NADP-isocitrate dehydrogenase was present, which appeared to be localised in the mitochondrion. No significant differences in maximal enzyme activities were observed except for NADP-isocitrate dehydrogenase, which showed decreased activity in production-phase mycelia. The results obtained support the scheme proposed by C.P. Kubicek for the intracellular organisation of citric acid formation but provide little evidence that this process is controlled at the level of the biosynthesis of any of the enzymes examined here.

Homocitrate synthase from Penicillium chrysogenum. Localization, purification of the cytosolic isoenzyme, and sensitivity to lysine.

Subcellular fractionation of cell-free extracts obtained by nitrogen cavitation showed that Penicillium chrysogenum Q176 contains a cytosolic as well as a mitochondrial homocitrate synthase activity. The cytosolic isoenzyme was purified about 500-fold, and its kinetic and molecular properties were investigated. Native homocitrate synthase shows a molecular mass of 155 +/- 10 kDa as determined by gel filtration and a pH of 4.9 +/- 0.1 as determined by chromatofocusing. The kinetic behaviour towards 2-oxoglutarate is hyperbolic, with Km = 2.2 mM; with respect to acetyl-CoA the enzyme shows sigmoidal saturation kinetics, with [S]0.5 = 41 microM and h = 2.6. The enzyme was inhibited strongly by L-lysine (Ki = 8 +/- 2 microM; 50% inhibition by 53 microM at 6 mM-2-oxoglutarate), competitively with 2-oxoglutarate, in protamine sulphate-treated and desalted cell-free extracts and in partially purified preparations. The extent of this inhibition was strongly pH-dependent. Both isoenzymes are equally susceptible to inhibition by lysine. The same inhibition pattern is shown by the enzyme from strain D6/1014A, which is a better producer of penicillin than strain Q176.

Lysine biosynthesis in Penicillium chrysogenum is regulated by feedback inhibition of alpha-aminoadipate reductase.

A partially purified preparation of alpha-aminoadipate reductase (EC 1.2.1.31) from Penicillium chrysogenum is competitively inhibited by lysine (Ki of 0.26 mM). Exogenous addition of 10 mM L-lysine to resting mycelia of P. chrysogenum increased the intracellular lysine pool concentration 2-fold, but decreased the incorporation of (6-14C)-alpha-aminoadipate into protein-bound lysine to a fifth. The distribution of radioactivity in the pathway metabolites alpha-aminoadipate, saccharopine and lysine was consistent with the assumption of a lysine sensitive enzyme step in vivo between alpha-aminoadipate and saccharopine. Hence lysine inhibition of alpha-aminoadipate reductase may be of physiologic importance.

alpha-Aminoadipate pool concentration and penicillin biosynthesis in strains of Penicillium chrysogenum.

Intracellular amino acid pools in four Penicillium chrysogenum strains, which differed in their ability to produce penicillin, were determined under conditions supporting growth without penicillin production and under conditions supporting penicillin production. A significant correlation between the rate of penicillin production and the intracellular concentration of alpha-aminoadipate was observed, which was not shown with any other amino acid in the pool. In replacement cultivation, penicillin production was stimulated by alpha-aminoadipate, but not by valine or cysteine. Exogenously added alpha-aminoadipate (2 or 3 mM) maximally stimulated penicillin synthesis in two strains of different productivity. Under these conditions intracellular concentrations of alpha-aminoadipate were comparable in the two strains in spite of the higher rate of penicillin production in the more productive strain. Results suggest that the lower penicillin titre of strain Q 176 is due to at least two factors: (i) the intracellular concentration of alpha-aminoadipate is insufficient to allow saturation of any enzyme which is rate limiting in the conversion of alpha-aminoadipate to penicillin and (ii) the level of an enzyme, which is rate limiting in the conversion of alpha-aminoadipate to penicillin, is lower in Q 176 (relative to strain D6/1014/A). Results suggest that the intracellular concentration of alpha-aminoadipate in strain D6/1014/A is sufficiently high to allow saturation of the rate-limiting penicillin biosynthetic enzyme in that strain. The basis of further correlation of intracellular alpha-aminoadipate concentration and penicillin titre among strains D6/1014/A, P2, and 389/3, the three highest penicillin producers studied here, remains to be established.(ABSTRACT TRUNCATED AT 250 WORDS)