Taxonomy, phylogeny and identification of Chaetomiaceae with emphasis on thermophilic species.

Chaetomiaceae comprises phenotypically diverse species, which impact biotechnology, the indoor environment and human health. Recent studies showed that most of the traditionally defined genera in Chaetomiaceae are highly polyphyletic. Many of these morphology-based genera, such as Chaetomium, Thielavia and Humicola, have been redefined using multigene phylogenetic analysis combined with morphology; however, a comprehensive taxonomic overview of the family is lacking. In addition, the phylogenetic relationship of thermophilic Chaetomiaceae species with non-thermophilic taxa in the family is largely unclear due to limited taxon sampling in previous studies. In this study, we provide an up-to-date overview on the taxonomy and phylogeny of genera and species belonging to Chaetomiaceae, including an extensive taxon sampling of thermophiles. A multigene phylogenetic analysis based on the ITS (internal transcribed spacers 1 and 2 including the 5.8S nrDNA), LSU (D1/D2 domains of the 28S nrDNA), rpb2 (partial RNA polymerase II second largest subunit gene) and tub2 (ß-tubulin gene) sequences was performed on 345 strains representing Chaetomiaceae and 58 strains of other families in Sordariales. Divergence times based on the multi-gene phylogeny were estimated as aid to determine the genera in the family. Genera were delimited following the criteria that a genus must be a statistically well-supported monophyletic clade in both the multigene phylogeny and molecular dating analysis, fall within a divergence time of over 27 million years ago, and be supported by ecological preference or phenotypic traits. Based on the results of the phylogeny and molecular dating analyses, combined with morphological characters and temperature-growth characteristics, 50 genera and 275 species are accepted in Chaetomiaceae. Among them, six new genera, six new species, 45 new combinations and three new names are proposed. The results demonstrate that the thermophilic species fall into seven genera (Melanocarpus, Mycothermus, Remersonia, Thermocarpiscus gen. nov., Thermochaetoides gen. nov., Thermothelomyces and Thermothielavioides). These genera cluster in six separate lineages, suggesting that thermophiles independently evolved at least six times within the family. A list of accepted genera and species in Chaetomiaceae, together with information on their MycoBank numbers, living ex-type strains and GenBank accession numbers to ITS, LSU, rpb2 and tub2 sequences is provided. Furthermore, we provide suggestions how to describe and identify Chaetomiaceae species. Taxonomic novelties: new genera: Parvomelanocarpus X.Wei Wang & Houbraken, Pseudohumicola X.Wei Wang, P.J. Han, F.Y. Bai & Houbraken, Tengochaeta X.Wei Wang & Houbraken, Thermocarpiscus X.Wei Wang & Houbraken, Thermochaetoides X.Wei Wang & Houbraken, Xanthiomyces X.Wei Wang & Houbraken; New species: Botryotrichum geniculatum X.Wei Wang, P.J. Han & F.Y. Bai, Chaetomium subaffine Sergejeva ex X.Wei Wang & Houbraken, Humicola hirsuta X.Wei Wang, P.J. Han & F.Y. Bai, Subramaniula latifusispora X.Wei Wang, P.J. Han & F.Y. Bai, Tengochaeta nigropilosa X.Wei Wang & Houbraken, Trichocladium tomentosum X.Wei Wang, P.J. Han & F.Y. Bai; New combinations: Achaetomiella gracilis (Udagawa) Houbraken, X.Wei Wang, P.J. Han & F.Y. Bai, Allocanariomyces americanus (Cañete-Gibas et al.) Cañete-Gibas, Wiederhold, X.Wei Wang & Houbraken, Amesia dreyfussii (Arx) X.Wei Wang & Houbraken, Amesia raii (G. Malhotra & Mukerji) X.Wei Wang & Houbraken, Arcopilus macrostiolatus (Stchigel et al.) X.Wei Wang & Houbraken, Arcopilus megasporus (Sörgel ex Seth) X.Wei Wang & Houbraken, Arcopilus purpurascens (Udagawa & Y. Sugiy.) X.Wei Wang & Houbraken, Arxotrichum deceptivum (Malloch & Benny) X.Wei Wang & Houbraken, Arxotrichum gangligerum (L.M. Ames) X.Wei Wang & Houbraken, Arxotrichum officinarum (M. Raza & L. Cai) X.Wei Wang & Houbraken, Arxotrichum piluliferoides (Udagawa & Y. Horie) X.Wei Wang & Houbraken, Arxotrichum repens (Guarro & Figueras) X.Wei Wang & Houbraken, Arxotrichum sinense (K.T. Chen) X.Wei Wang & Houbraken, Botryotrichum inquinatum (Udagawa & S. Ueda) X.Wei Wang & Houbraken, Botryotrichum retardatum (A. Carter & R.S. Khan) X.Wei Wang & Houbraken, Botryotrichum trichorobustum (Seth) X.Wei Wang & Houbraken, Botryotrichum vitellinum (A. Carter) X.Wei Wang & Houbraken, Collariella anguipilia (L.M. Ames) X.Wei Wang & Houbraken, Collariella hexagonospora (A. Carter & Malloch) X.Wei Wang & Houbraken, Collariella pachypodioides (L.M. Ames) X.Wei Wang & Houbraken, Ovatospora amygdalispora (Udagawa & T. Muroi) X.Wei Wang & Houbraken, Ovatospora angularis (Yu Zhang & L. Cai) X.Wei Wang & Houbraken, Parachaetomium biporatum (Cano & Guarro) X.Wei Wang & Houbraken, Parachaetomium hispanicum (Guarro & Arx) X.Wei Wang & Houbraken, Parachaetomium inaequale (Pidopl. et al.) X.Wei Wang & Houbraken, Parachaetomium longiciliatum (Yu Zhang & L. Cai) X.Wei Wang & Houbraken, Parachaetomium mareoticum (Besada & Yusef) X.Wei Wang & Houbraken, Parachaetomium muelleri (Arx) X.Wei Wang & Houbraken, Parachaetomium multispirale (A. Carter et al.) X.Wei Wang & Houbraken, Parachaetomium perlucidum (Sergejeva) X.Wei Wang & Houbraken, Parachaetomium subspirilliferum (Sergejeva) X.Wei Wang & Houbraken, Parathielavia coactilis (Nicot) X.Wei Wang & Houbraken, Parvomelanocarpus tardus (X.Wei Wang & Samson) X.Wei Wang & Houbraken, Parvomelanocarpus thermophilus (Abdullah & Al-Bader) X.Wei Wang & Houbraken, Pseudohumicola atrobrunnea (X.Wei Wang et al.) X.Wei Wang, P.J. Han, F.Y. Bai & Houbraken, Pseudohumicola pulvericola (X.Wei Wang et al.) X.Wei Wang, P.J. Han, F.Y. Bai & Houbraken, Pseudohumicola semispiralis (Udagawa & Cain) X.Wei Wang, P.J. Han, F.Y. Bai & Houbraken, Pseudohumicola subspiralis (Chivers) X.Wei Wang, P.J. Han, F.Y. Bai & Houbraken, Staphylotrichum koreanum (Hyang B. Lee & T.T.T. Nguyen) X.Wei Wang & Houbraken, Staphylotrichum limonisporum (Z.F. Zhang & L. Cai) X.Wei Wang & Houbraken, Subramaniula lateralis (Yu Zhang & L. Cai) X.Wei Wang & Houbraken, Thermocarpiscus australiensis (Tansey & M.A. Jack) X.Wei Wang & Houbraken, Thermochaetoides dissita (Cooney & R. Emers.) X.Wei Wang & Houbraken, Thermochaetoides thermophila (La Touche) X.Wei Wang & Houbraken, Xanthiomyces spinosus (Chivers) X.Wei Wang & Houbraken; New names: Chaetomium neoglobosporum X.Wei Wang & Houbraken, Thermothelomyces fergusii X.Wei Wang & Houbraken, Thermothelomyces myriococcoides X.Wei Wang & Houbraken; Lecto- and / or epi-typifications (basionyms): Botryoderma rostratum Papendorf & H.P. Upadhyay, Botryotrichum piluliferum Sacc. & Marchal, Chaetomium carinthiacum Sörgel, Thielavia heterothallica Klopotek. Citation: Wang XW, Han PJ, Bai FY, Luo A, Bensch K, Meijer M, Kraak B, Han DY, Sun BD, Crous PW, Houbraken J (2022). Taxonomy, phylogeny and identification of Chaetomiaceae with emphasis on thermophilic species. Studies in Mycology 101: 121-243. doi: 10.3114/sim.2022.101.03.

Phylogenetic re-evaluation of Thielavia with the introduction of a new family Podosporaceae.

The genus Thielavia is morphologically defined by having non-ostiolate ascomata with a thin peridium composed of textura epidermoidea, and smooth, single-celled, pigmented ascospores with one germ pore. Thielavia is typified with Th. basicola that grows in close association with a hyphomycete which was traditionally identified as Thielaviopsis basicola. Besides Th. basicola exhibiting the mycoparasitic nature, the majority of the described Thielavia species are from soil, and some have economic and ecological importance. Unfortunately, no living type material of Th. basicola exists, hindering a proper understanding of the classification of Thielavia. Therefore, Thielavia basicola was neotypified by material of a mycoparasite presenting the same ecology and morphology as described in the original description. We subsequently performed a multi-gene phylogenetic analyses (rpb2, tub2, ITS and LSU) to resolve the phylogenetic relationships of the species currently recognised in Thielavia. Our results demonstrate that Thielavia is highly polyphyletic, being related to three family-level lineages in two orders. The redefined genus Thielavia is restricted to its type species, Th. basicola, which belongs to the Ceratostomataceae (Melanosporales) and its host is demonstrated to be Berkeleyomyces rouxiae, one of the two species in the "Thielaviopsis basicola" species complex. The new family Podosporaceae is sister to the Chaetomiaceae in the Sordariales and accommodates the re-defined genera Podospora, Trangularia and Cladorrhinum, with the last genus including two former Thielavia species (Th. hyalocarpa and Th. intermedia). This family also includes the genetic model species Podospora anserina, which was combined in Triangularia (as Triangularia anserina). The remaining Thielavia species fall in ten unrelated clades in the Chaetomiaceae, leading to the proposal of nine new genera (Carteria, Chrysanthotrichum, Condenascus, Hyalosphaerella, Microthielavia, Parathielavia, Pseudothielavia, Stolonocarpus and Thermothielavioides). The genus Canariomyces is transferred from Microascaceae (Microascales) to Chaetomiaceae based on its type species Can. notabilis. Canariomyces is closely related to the human-pathogenic genus Madurella, and includes three thielavia-like species and one novel species. Three monotypic genera with a chaetomium-like morph (Brachychaeta, Chrysocorona and Floropilus) are introduced to better resolve the Chaetomiaceae and the thielavia-like species in the family. Chrysocorona lucknowensis and Brachychaeta variospora are closely related to Acrophialophora and three newly introduced genera containing thielavia-like species; Floropilus chiversii is closely related to the industrially important and thermophilic species Thermothielavioides terrestris (syn. Th. terrestris). This study shows that the thielavia-like morph is a homoplastic form that originates from several separate evolutionary events. Furthermore, our results provide new insights into the taxonomy of Sordariales and the polyphyletic Lasiosphaeriaceae.

Redefining Humicola sensu stricto and related genera in the Chaetomiaceae.

The traditional concept of the genus Humicola includes species that produce pigmented, thick-walled and single-celled spores laterally or terminally on hyphae or minimally differentiated conidiophores. More than 50 species have been described in the genus. Species commonly occur in soil, indoor environments, and compost habitats. The taxonomy of Humicola and morphologically similar genera is poorly understood in modern terms. Based on a four-locus phylogeny, the morphological concept of Humicola proved to be polyphyletic. The type of Humicola, H. fuscoatra, belongs to the Chaetomiaceae. In the Chaetomiaceae, species producing humicola-like thick-walled spores are distributed among four lineages: Humicola sensu stricto, Mycothermus, Staphylotrichum, and Trichocladium. In our revised concept of Humicola, asexual and sexually reproducing species both occur. The re-defined Humicola contains 24 species (seven new and thirteen new combinations), which are described and illustrated in this study. The species in this genus produce conidia that are lateral, intercalary or terminal on/in hyphae, and conidiophores are not formed or are minimally developed (micronematous). The ascospores of sexual Humicola species are limoniform to quadrangular in face view and bilaterally flattened with one apical germ pore. Seven species are accepted in Staphylotrichum (four new species, one new combination). Thick-walled conidia of Staphylotrichum species usually arise either from hyphae (micronematous) or from apically branched, seta-like conidiophores (macronematous). The sexual morph represented by Staphylotrichum longicolleum (= Chaetomium longicolleum) produces ascomata with long necks composed of a fused basal part of the terminal hairs, and ascospores that are broad limoniform to nearly globose, bilaterally flattened, with an apical germ pore. The Trichocladium lineage has a high morphological diversity in both asexual and sexual structures. Phylogenetic analysis revealed four subclades in this lineage. However, these subclades are genetically closely related, and no distinctive phenotypic characters are linked to any of them. Fourteen species are accepted in Trichocladium, including one new species, twelve new combinations. The type species of Gilmaniella, G. humicola, belongs to the polyphyletic family Lasiosphaeriaceae (Sordariales), but G. macrospora phylogenetically belongs to Trichocladium. The thermophilic genus Mycothermus and the type species My. thermophilum are validated, and one new Mycothermus species is described. Phylogenetic analyses show that Remersonia, another thermophilic genus, is sister to Mycothermus and two species are known, including one new species. Thermomyces verrucosus produces humicola-like conidia and is transferred to Botryotrichum based on phylogenetic affinities. This study is a first attempt to establish an inclusive modern classification of Humicola and humicola-like genera of the Chaetomiaceae. More research is needed to determine the phylogenetic relationships of "humicola"-like species outside the Chaetomiaceae.

Polyketides with different post-modifications from desert endophytic fungus Paraphoma sp.

Three new polyketides 4,6,8-trihydroxy-5-methyl-3,4-dihydronaphthalen-1(2H)-one (1), 5,7-dihydroxy-3-(1-hydroxyethyl)-3,4-dimethylisobenzofuran-1(3H)-one (2) and 1-(4-hydroxy-6-methoxy-1,7-dimethyl-3-oxo-1,3-dihydroisobenzofuran-1-yl) ethyl acetate (3) together with seven known analogues (4-10) were isolated from desert endophytic fungus Paraphoma sp. The structures of these compounds were elucidated by analysis of NMR data. The absolute configuration of (1-3) was established on the basis of CD experiments. The possible biosynthetic pathway of compounds (1-10) was suggested, which implied that these secondary metabolites might be originated from polyketide biosynthesis with different post-modification reactions. Compounds 2, and 5-8 were evaluated for bioactivities against plant pathogen A. solani, whereas none of them displayed any biological effects. In addition, compounds 1, 2 and 5-10 were also tested for cytotoxic activities against three human cancer cell lines (HepG2 cells, MCF-7 cells and Hela cells) without biological effects.

New Talaromyces species from indoor environments in China.

Talaromyces contains both asexual and sexually reproducing species. This genus is divided in seven sections and currently has 105 accepted species. In this study we investigated the Talaromyces isolates that were obtained during a study of indoor air collected in Beijing, China. These indoor Talaromyces strains are resolved in four sections, seven of them are identified as T. islandicus, T. aurantiacus, T. siamensis and T. albobiverticillius according to BenA sequences, while 14 isolates have divergent sequences and are described here as nine new species. The new species are placed in four sections, namely sections Helici, Islandici, Talaromyces and Trachyspermi. They are described based on sequence data (ITS, BenA, CaM and RPB2) in combination with phenotypic and extrolite characters. Morphological descriptions and notes for distinguishing similar species are provided for each new species. The recently described T. rubrifaciens is synonymised with T. albobiverticillius based on presented phylogenetic results.

Aspergillus section Nidulantes (formerly Emericella): Polyphasic taxonomy, chemistry and biology.

Aspergillus section Nidulantes includes species with striking morphological characters, such as biseriate conidiophores with brown-pigmented stipes, and if present, the production of ascomata embedded in masses of Hülle cells with often reddish brown ascospores. The majority of species in this section have a sexual state, which were named Emericella in the dual name nomenclature system. In the present study, strains belonging to subgenus Nidulantes were subjected to multilocus molecular phylogenetic analyses using internal transcribed spacer region (ITS), partial ß-tubulin (BenA), calmodulin (CaM) and RNA polymerase II second largest subunit (RPB2) sequences. Nine sections are accepted in subgenus Nidulantes including the new section Cavernicolus. A polyphasic approach using morphological characters, extrolites, physiological characters and phylogeny was applied to investigate the taxonomy of section Nidulantes. Based on this approach, section Nidulantes is subdivided in seven clades and 65 species, and 10 species are described here as new. Morphological characters including colour, shape, size, and ornamentation of ascospores, shape and size of conidia and vesicles, growth temperatures are important for identifying species. Many species of section Nidulantes produce the carcinogenic mycotoxin sterigmatocystin. The most important mycotoxins in Aspergillus section Nidulantes are aflatoxins, sterigmatocystin, emestrin, fumitremorgins, asteltoxins, and paxillin while other extrolites are useful drugs or drug lead candidates such as echinocandins, mulundocandins, calbistrins, varitriols, variecolins and terrain. Aflatoxin B1 is produced by four species: A. astellatus, A. miraensis, A. olivicola, and A. venezuelensis.

Dominant factor determining the conduction-type of nitrogen-doped ZnO film.

Nitrogen-doped zinc oxide (ZnO) film has been grown by molecular beam epitaxy. The as-grown sample showed p-type conduction with a hole concentration of 3.1 x 10(17) cm(-3). After an annealing process in O2 at 600 degrees C for 30 min, p-type conduction was still remained, and the hole concentration of the film decreased to 6.8 x 10(16) cm(-3). Secondary ion mass spectroscopy revealed that the concentration of both nitrogen and hydrogen decreased after the annealing process. It is demonstrated that the intrinsic compensation source has been decreased after the annealing process. Because the variation trend of the hole concentration in the ZnO:N film is opposite to that of hydrogen and intrinsic defects, but in good accordance with nitrogen, the extrinsically substituted nitrogen (N(o)) should be the dominant factor that determines the conduction-type of the ZnO:N film.

Early, sustained efficacy of adeno-associated virus vector-mediated gene therapy in glycogen storage disease type Ia.

The deficiency of glucose-6-phosphatase (G6Pase) underlies life-threatening hypoglycemia and growth retardation in glycogen storage disease type Ia (GSD-Ia). An adeno-associated virus (AAV) vector encoding G6Pase was pseudotyped as AAV8 and administered to 2-week-old GSD-Ia mice (n = 9). Median survival was prolonged to 7 months following vector administration, in contrast to untreated GSD-Ia mice that survived for only 2 weeks. Although GSD-Ia mice were initially growth-retarded, treated mice increased fourfold in weight to normal size. Blood glucose was partially corrected by 2 weeks following treatment, whereas blood cholesterol normalized. Glucose-6-phosphatase activity was partially corrected to 25% of the normal level at 7 months of age in treated mice, and blood glucose during fasting remained lower in treated, affected mice than in normal mice. Glycogen storage was partially corrected in the liver by 2 weeks following treatment, but reaccumulated to pre-treatment levels by 7 months old (m.o.). Vector genome DNA decreased between 3 days and 3 weeks in the liver following vector administration, mainly through the loss of single-stranded genomes; however, double-stranded vector genomes were more stable. Although CD8+ lymphocytic infiltrates were present in the liver, partial biochemical correction was sustained at 7 m.o. The development of efficacious AAV vector-mediated gene therapy could significantly reduce the impact of long-term complications in GSD-Ia, including hypoglycemia, hyperlipidemia and growth failure.

Freeze-substitution and low-temperature embedding of dairy products for transmission electron microscopy.

Dairy products are comprised largely of fat, air and water, which makes it difficult to preserve their ultrastructure for electron microscopy. Keeping the samples frozen throughout fixation and embedding protects the structure and distribution of the components of emulsions and foams. Therefore, dairy products were freeze-substituted and embedded at low temperature (-20 degrees C) to prepare them for transmission electron microscopy. Whipped cream, ice cream mix and dairy/non-dairy mixed systems were frozen by plunging in propane, at its boiling point (-187 degrees C). Ice cream, because it is already frozen, was fractured into 1-mm3 pieces in liquid nitrogen and then added to frozen fixative (-196 degrees C). Fixative solution consisted of glutaraldehyde, osmium tetroxide and uranyl acetate dissolved in either methanol or acetone. When material was to be stained after sectioning the fixative was limited to glutaraldehyde in methanol. The temperature was increased step-wise from -80 to -20 degrees C. Solvent was replaced with resin; the polar resin Lowicryl HM4, the non-polar resin Lowicryl HM20, LR White and LR Gold were tested. Samples were embedded and polymerized at -20 degrees C using ultraviolet light to cross-link the resin. Methanol proved to be the most effective solvent for substituting the ice; the hydrophobic resin Lowicryl HM20 was the most effective resin for retaining fat structure following osmium fixation.