Taxonomic revision of Bisifusarium (Nectriaceae).

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 (β-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.

The genus Exophiala is an anamorphic ascomycete fungus in the family Herpotrichiellaceae of the order Chaetothyriales. Exophiala species have been classified as polymorphic black yeast-like fungi. Prior to this study, 63 species had been validated, published, and accepted into this genus. Exophiala species are known to be distributed worldwide and have been isolated in various habitats around the world. Several Exophiala species have been identified as potential agents of human and animal mycoses. However, in some studies, Exophiala species have been used in agriculture and biotechnological applications. Here, we provide a brief review of the diversity, distribution, and taxonomy of Exophiala through an overview of the recently published literature. Moreover, four new Exophiala species were isolated from rocks that were collected from natural forests located in northern Thailand. Herein, we introduce these species as E. lamphunensis, E. lapidea, E. saxicola, and E. siamensis. The identification of these species was based on a combination of morphological characteristics and molecular analyses. Multi-gene phylogenetic analyses of a combination of the internal transcribed spacer (ITS) and small subunit (nrSSU) of ribosomal DNA, along with the translation elongation factor (tef), partial β-tubulin (tub), and actin (act) genes support that these four new species are distinct from previously known species of Exophiala. A full description, illustrations, and a phylogenetic tree showing the position of four new species are provided.

The variability within Aspergillus fumigatus Fresenius and related species was examined using macro-, micro-morphology, growth temperature regimes and extrolite patterns. In addition, DNA analyses including partial β-tubulin, calmodulin and actin gene sequences were used. Detailed examination of strains, considered as A. fumigatus earlier, showed that they could be divided into four groups including A. fumigatus sensu stricto, A. lentulus and two new species. The intraspecific genetic variability within A. fumigatus sensu stricto was low, the sequence differences among 23 strains of the species was at most two bases in each partial β-tubulin and calmodulin gene. However, intraspecific morphological diversity within the species was high and delineation of the species was equivocal. Therefore, β-tubulin and calmodulin gene sequences could be critical determinants for the delineation of the A. fumigatus sensu stricto species. A. lentulus including isolates from clinical origin, Korean soil and from a dolphin clustered into an isolated group based on β-tubulin, calmodulin and actin gene sequences, differing from A. fumigatus by morphological characters, growth temperature and extrolite profile. A. lentulus produces the extrolites auranthine, cyclopiazonic acid, a dimeric indole of unknown structure, neosartorin, some pyripyropens, terrein and some tryptoquivalins and tryptoquivalons. Two pair of isolates (CBS 117194, 117186 and 117520, 117519) clustered into separate groups from A. fumigatus and the other Aspergillus section Fumigati species, including the teleomorph Neosartorya, are proposed as two new species. A. fumigatiaffinis spec. nov. produces the extrolites auranthine, cycloechinulin, helvolic acid, neosartorin, palitantin, pyripyropens, tryptoquivalins and tryptoquivalons, and A. novofumigatus spec. nov. produces the extrolites cycloechinuline, helvolic acid, neosartorin, palitantin and terrein.

Blueberry (Vaccinium spp.) has been cultivated in more than 20 provinces in China as reported in 2015. In July 2016, a root rot disease of blueberry causing plant death was found in Qujing (24°49′N; 103°59′E), Yunnan Province. Approximately 5% of plants in the field showed symptoms characterized by leaf chlorosis, black rot on roots and stems, and finally the whole plant wilted in about 20 days. In order to identify the causal agent of this disease, typical disease samples were collected. Twenty infected root tissues were sterilized and put on potato dextrose agar (PDA) at 25°C (Fang 1998) and nine fungal isolates were obtained. The isolate YNQJ01Aa was chosen for further testing. Its colony on PDA was white to pale yellow. The conidia were collected from SNA colony after growing for 10 days. Conidia presented with one to three septa (mostly one septa). Fifty conidia from one isolate measured 59.37 to 71.33 × 5.93 to 7.91 μm (avg. 62.25 × 6.22 µm). Ascospores were produced in perithecia on PDA and were 30.16 to 38.89 × 5.78 to 7.76 µm (avg. 34.97 × 6.52 µm). By DNA sequencing of the internal transcribed spacer (ITS) rDNA region (GenBank accession no. MF785081), the identity of the isolate (YNQJ01Aa) was confirmed to be the genus Calonectria (99% homologous to several Calonectria spp. strains). The partial β-tubulin (TUB) gene sequence was also analyzed (MF785082) and the result of a BLAST search showed that it was 100% homologous to C. ilicicola strain (CBS 190.50). Further species identification was analyzed by phylogenic tree based on TUB (partial β-tubulin gene, T1/Bt2b), tef1 (translation elongation factor 1 alpha gene, EF1-728F/EF1-986R), and HIS3 (histone 3 gene, H3-1a/H3-1b) (Chen et al. 2011; Crous 2002). Sequences obtained were deposited in GenBank (TUB: MF785082, tef1: MF785083, HIS3: MF785084). Alignments were made using CLUSTAL X v.2.0, and a maximum likelihood phylogram was generated by MEGA v.7.0. Isolate YNQJ01Aa clustered together with C. ilicicola reference strain CBS190.50 (98% bootstrap). Three healthy 1-year-old ‘O’Neal’ blueberry plants were used to conduct pathogenicity tests by wound inoculation. Each plant was inoculated with 10 ml of conidial suspension (1 × 10⁵ conidia/ml) by the root irrigation method. Three plants were inoculated with the same amount of sterile water as control. All blueberry plants were incubated at 25°C. After 20 days, the test plants showed typical symptom of black root rot and finally the whole plant wilted and died, while the control plants remained healthy. The same colonial fungus was reisolated. This is the first report of C. ilicicola (anamorph Cylindrocladium parasiticum) infecting blueberry in China. Root rot disease can be difficult to detect early and therefore it is important to identify the etiology to develop appropriate disease management strategies. This fungus was previously reported on Medicago sativa in China (Pei et al. 2015).

During a study on the mycobiota of brazil nuts (Bertholletia excelsa) in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial β-tubulin, calmodulin and ITS sequences to characterize this taxon. A. bertholletius is represented by nineteen isolates from samples of brazil nuts at various stages of production and soil close to Bertholletia excelsa trees. The following extrolites were produced by this species: aflavinin, cyclopiazonic acid, kojic acid, tenuazonic acid and ustilaginoidin C. Phylogenetic analysis using partial β-tubulin and camodulin gene sequences showed that A. bertholletius represents a new phylogenetic clade in Aspergillus section Flavi. The type strain of A. bertholletius is CCT 7615 ( = ITAL 270/06 = IBT 29228).

Maize (Zea mays L.) is one of the most important crops in China. Ear rot is a serious disease that often leads to a decline in maize yield and quality and is a hazard to human and livestock health owing to potential mycotoxin contamination of grain. The incidence of maize ear rot usually ranges from 20 to 40% at different fields, even reaching as high as 70% at certain plots in Chongqing and surrounding areas (Zhou et al. 2018). In August 2016, a survey to determine population composition and distribution of Fusarium spp. causing maize ear rot was conducted in 22 counties in Chongqing. Maize ears with white, pink, or salmon-colored mold were collected from fields for further analysis. Symptomatic kernels were soaked in 2% sodium hypochlorite solution for 3 min, rinsed with sterile water three times, placed on acidified potato dextrose agar (PDA), and incubated at 26 ± 1°C for 3 to 5 days. The newly grown-out fungal colonies displaying morphological characteristics of Fusarium spp. were transferred onto fresh PDA and Spezieller Nahrstoffarmer agar and purified by the single-spore isolation method (Yang et al. 2008). Fusarium spp. were identified based on morphological characteristics (Leslie and Summerell 2006) and sequence analysis of the translation elongation factor-1α (TEF-1α), β-tubulin, and mitochondrial small subunit (mtSSU) genes (Jiang et al. 2018). The predominant Fusarium spp. were Fusarium verticillioides, F. proliferatum, and F. meridionale, which were the main causal agents of maize ear rot in these areas. In addition, morphological characteristics of several isolates from Beibei, Jiangjin, Xiushan, Rongchang, Kaixian, and Wanzhou Districts were found to be identical to those of Fusarium fujikuroi Nirenberg, which grouped into the Gibberella fujikuroi species complex. Colonies on PDA produced abundant white aerial mycelia initially, but the mycelia became pale pinkish and dark violet with age. Microconidia were abundant and small in size (4.9 to 11.3 × 2.2 to 3.2 µm), colorless, oval to club shaped, and 0- to 1-septate. Macroconidia were slender, almost straight, sickle shaped, medium length, with three to five septa (17.1 to 39.2 × 2.6 to 3.3 µm). No chlamydospores were observed. Identity of the fungus was also confirmed by sequence comparison of the partial TEF-1α, β-tubulin, and mtSSU genes of two representative isolates. The resulting sequences of the TEF-1α, β-tubulin, and mtSSU genes showed 100, 100, and 99% sequence identity to those of F. fujikuroi (GenBank accession nos. MH263736.1, MH263737.1, and MF984420.1, respectively). DNA sequences of partial TEF-1α, β-tubulin, and mtSSU genes from two representative isolates (FCQD90 and FCQD97) were deposited in GenBank under accession numbers MK328875 and MK328876, MK385626 and MK385627, and MK385628 and MK385629, respectively. A pathogenicity test was performed on maize inbred lines B73 and Ye 107. Four days after silk emergence, 2 ml of conidial suspension (1 × 10⁶ conidia /ml) of each isolate was injected into each of 10 maize ears through the silk channel. Control plants were inoculated with sterile distilled water. The typical Fusarium ear rot symptom (white or white-pink mold) was observed on inoculated ears and the infected kernels. No symptom developed on the water-inoculated controls. F. fujikuroi was reisolated from the symptomatic ears but not from the control. F. fujikuroi was previously reported associated with seedling damping-off on soybean, stem wilt of Canna edulis, and root rot of Reineckia carnea (Jiang et al. 2018; Pedrozo et al. 2015; Sun et al. 2018). To our knowledge, this is the first report of F. fujikuroi causing maize ear rot in China, and this disease should be paid sufficient attention owing to a serious risk of mycotoxin contamination in maize.

Coprophilous fungi are saprotrophic organisms that show great diversity, mainly on herbivore dung. The physico-chemical characteristics of this peculiar substrate combined with the high level of fungal adaptation to different environmental conditions offer the perfect setting for discovering new taxa. This study focused on the species diversity of penicillium-like fungi isolated mainly from herbivore dung collected at different Spanish locations. From 130 samples, a total of 104 isolates were obtained, and 48 species were identified. Preliminary identifications were based on morphology and partial β-tubulin (tub2) gene sequences. Putative new taxa were characterized by a multi-gene sequencing analysis testing the tub2, the internal transcribed spacer rDNA (ITS), calmodulin (cmdA), and RNA polymerase II second largest subunit (rpb2) genes, and a detailed phenotypic study. Using this polyphasic approach and following the genealogical concordance phylogenetic species recognition (GCPSR) method, we propose the new genera Penicillago (for Penicillium nodositatum) and Pseudopenicillium (for Penicillium megasporum and P. giganteum) in the family Aspergillaceae, and 11 new species, including seven Penicillium, three Talaromyces and one Pseudopenicillium. A lectotype and epitype are designed for Penicillium nodositatum. Our results show that the species diversity of penicillium-like fungi on herbivore dung has not been widely studied and that this substrate seems to be a good reservoir of interesting Eurotialean fungi.

Stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is one of the most devastating diseases of wheat (Triticum spp.) worldwide. Indian isolates were characterised based on their phenotypic reaction on differential hosts carrying different Yr genes. Based on virulence/avirulence structure, isolates were characterised into ten different pathotypes viz. 70S0-2, 67S64, 70S4, 66S0, 70S64, 66S64-1, 38S102, 47S102, 46S119, and 78S84. These Indian pathotypes of P. striiformis f. sp. tritici and 38 pathotypes of other rust species (P. graminis tritici and P. triticina) were used in this study to analyze their molecular phylogenetic relationship. The nucleotides of rDNA-ITS, partial β-tubulin and ketopantoate reductase genes of all the pathotypes were sequenced directly after PCR. Based on sequence data of rDNA-ITS and β-tubulin, three phylogenetic groups corresponding to three different species of Puccinia were obtained. Asian isolates formed a distinct evolutionary lineage than from those derived from USA. The sequence similarity of Indian pathotypes with other Asian (China and Iran) isolates indicated the same origin of pathotypes. The results will allow rapid identification of Indian P.striiformis f. sp. tritici pathotypes causing stripe rust in wheat, assist in making predictions regarding potential rust pathotypes, and identifying sources of resistance to the disease in advance.

Isolates of Venturia species isolated from pear in Japan, China, Taiwan and Israel were used in this study to analyze their molecular phylogenetic relationship. The nucleotides of rDNA-ITS, partial β-tubulin and elongation factor 1α genes were sequenced directly after PCR. Based on these sequence data two phylogenetic groups could be distinguished. Isolates collected from Asian pears such as Japanese and Chinese pears formed a distinct evolutionary lineage from those derived from European and Syrian pears. This result corroborated the early taxonomic separation of V. nashicola from V. pirina. In addition, trees from single-locus data sets and the combined data set showed that all isolates of V. nashicola were included in a monophyletic group and representative isolates of five pathological races originating from different locations and cultivars formed a single lineage. In contrast, two distinct evolutionary lineages were revealed in V. pirina and isolates of five races were scattered in two lineages. Israeli isolates of race 2 as well as two Japanese isolates of V. pirina formed a distinct lineage from other isolates of this species, while other Israeli isolates belonging to races 1, 3, 4, and 5 were closely related to each other and formed another lineage. It was indicated that the evolution of pathological races in V. nashicola might have occurred relatively recently as compared with V. pirina.

Phylogeny of bipolar Cladonia arbuscula and Cladonia mitis (Lecanorales, Euascomycetes)

Pestalotiopsis revisited

During the continuous investigation into inconspicuous xylarialean species in northern Thailand, two amphisphaeria-like taxa were found in samples collected between July and October 2022. Both taxa have solitary, immersed ascomata with a two-layered peridium and unitunicate asci that are morphologically similar to Amphisphaeria species. Macro–micro morphological comparisons with related taxa and multigene phylogenetic analyses based on combined internal transcribed spacer (ITS), partial 28S large subunit rDNA (LSU), partial RNA polymerase II second largest subunit (rpb2), and partial β-tubulin (tub2) sequences revealed that the two taxa are novel to science. Amphisphaeria chiangmaiensis sp. nov. is distinguished from known taxa by having larger ascomata, a J- apical ring in Melzer’s reagent, and hyaline ascospores. Amphisphaeria hydei sp. nov. differs from its sister taxa in having larger ascomata, a longer ostiolar canal, and longer asci. Morphological descriptions, illustrations, and phylogenetic analyses of two novel taxa are provided herein.

Leaf blight is a common disease affecting Sansevieria trifasciata in many countries, including Malaysia. In the present study, Fusarium isolates were consistently recovered from the diseased leaves collected from various locations throughout the country. Based on morphology and multigene phylogenetic analysis using mitochondrial small subunit ( mtSSU ), intergenic spacer region (IGS) and translation elongation factor 1‐α ( TEF1‐α ) gene sequences, seven Fusarium species were identified, with F. oxysporum being the most prevalent (67.6%) among 34 isolates. Pathogenicity tests resulted in the discovery of pathogenic isolates that belonged to F. oxysporum , F. proliferatum , and F. pseudocircinatum , whereas all isolates of F. brachygibbosum , F. concentricum , F. mangiferae , and F. solani were nonpathogenic. The results suggest that several Fusarium species are accountable for causing disease on S. trifasciata in Malaysia.

First report of Alternaria arborescens causing white spot disease on Chinese chive in China

Paraboeremia was recently introduced for a distinct lineage in the family Didymellaceae. Currently, three species are included, i.e. P. adianticola, P. putaminum and P. selaginellae, all of which are plant pathogens. Paraboeremia is morphologically similar to Phoma but phylogenetically distinct. In this paper, three new species, i.e. Paraboeremia camelliae isolated from Camellia sp., P. litseae from Litsea sp., and P. oligotrophica from cave limestone, are described, illustrated and compared with closely related taxa. Phylogenetic analysis based on the multi-locus sequences of the internal transcribed spacer regions 1 and 2 and 5.8S nuclear ribosomal RNA gene (ITS), partial large subunit 28S nrDNA region (LSU), partial β-tubulin (TUB2) gene and RNA polymerase II (RPB2) gene regions confirmed the distinction of these species in Paraboeremia. These three new species were discovered from habitats and hosts that are previously unknown from this genus.