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Grapevine trunk diseases (GTDs), caused by a complex of fungi, are a global threat to vineyard longevity. Identifying the fungi in each growing region is crucial for interpreting symptoms and under...

In the version of this Perspective originally published, the author Vincent Robert’s affiliation with the Westerdijk Fungal Biodiversity Institute was mistakenly omitted. In addition, the author Marco Thines’s affiliation with the Senckenberg Biodiversity and Climate Research Centre was mistakenly omitted. Vincent Robert’s affiliation has now been corrected to: Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy; and Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands. Marco Thines’s affiliation has now been corrected to: International Commission on the Taxonomy of Fungi (ICTF), https://www. fungaltaxonomy.org/; Institute of Ecology, Evolution and Diversity, Goethe University, Frankfurt (Main), Germany; and Senckenberg Biodiversity and Climate Research Centre, Frankfurt (Main), Germany.

The identification and proper naming of microfungi, in particular plant, animal and human pathogens, remains challenging. Molecular identification is becoming the default approach for many fungal groups, and environmental metabarcoding is contributing an increasing amount of sequence data documenting fungal diversity on a global scale. This includes lineages represented only by sequence data. At present, these taxa cannot be formally described under the current nomenclature rules. By considering approaches used in bacterial taxonomy, we propose solutions for the nomenclature of taxa known only from sequences to facilitate consistent reporting and communication in the literature and public sequence repositories.

High-throughput sequencing technologies using amplicon approaches have changed the way that studies investigating fungal distribution are undertaken. These powerful and time-efficient technologies have the potential for the first time to accurately map fungal distributions across landscapes or changes in diversity across ecological or biological gradients of interest. There is no requirement for a fungus to form a fruiting body to be detected, and both culturable and nonculturable organisms can be detected. Here we use high-throughput amplicon sequencing from bulk DNA extracts to test the impact that biases associated with culture-based methods had on an earlier study that compared the influence of site and host on fungal diversity in Nothofagaceae forests in New Zealand. Both detection methods sampled tissue from the same set of symptomless, living leaves. We found that both the culturing and high-throughput approaches show that host is a stronger driver of fungal community structure than site, but that both methods have some taxonomic biases. We also found that the individual trees selected for high-throughput sampling can impact the alpha-diversity detected and through this could potentially affect subsequent analyses based on a comparison of this diversity.

Fungal taxonomy and classification, and fungal identification tools, are increasingly based on DNA sequencing technologies. In contrast, many historical records of fungi are based on morphologically identified specimens. Scientific collections of specimens or living cultures provide a resource to enable these early records to be genetically validated using modern techniques. This project uses a set of cultures deposited prior to 2008 in the International Collection of Microorganisms from Plants (ICMP) culture collection as Phoma, a genus of high biosecurity importance that is notoriously difficult to identify accurately using morphological characters. Of the 265 cultures sequenced, 23% had been misidentified. These misidentifications mean that six species previously thought to be present in New Zealand probably do not occur there. Fifty specimens had not previously been identified to species level; amongst these were eight species newly reported from New Zealand. Genetic validation projects such as this, using existing scientific collections, are essential for maintaining a robust global biosecurity system.

Species- and population-specific differences in fungicide resistance and aggressiveness within Botrytis makes basic data on genetic diversity important for understanding disease caused by this fungus. Genetic diversity of Botrytis was surveyed between 2008 and 2012 from grapes from five New Zealand wine-growing regions. A total of 1226 isolates were gathered from symptomless flower buds at the start of the growing season and 1331 isolates from diseased fruit at harvest. Two species were found, B. cinerea and B. pseudocinerea. Botrytis pseudocinerea was common in both Auckland vineyards sampled, and infrequent elsewhere. However, even in Auckland, it was rarely isolated from diseased fruit. The presence of the Boty and Flipper transposons was assessed. Isolates with all four transposon states (Boty only, Flipper only, both Boty and Flipper, no transposons) were found for both species. Both vineyards in the Auckland region had high numbers of Flipper-only isolates at flowering; both vineyards from the Waipara region had high numbers of Boty-only isolates at flowering. Most isolates from diseased fruit at harvest contained both transposons. These observations suggest that B. pseudocinerea, and isolates with one or both of the transposons missing, may be less aggressive than B. cinerea, or than isolates with both transposons present. Two clades were resolved within B. pseudocinerea, only one of which has been reported from European vineyards. Phylogenetic diversity within B. cinerea in New Zealand was similar to that known from Europe, including isolates that appear to match Botrytis ‘Group S’. The taxonomic implications of this genetic diversity are discussed.

Se evaluaron seis regiones de ADN como posibles códigos de barras de ADN para Hongos, el segundo reino más grande de vida eucariota, por un consorcio multinacional y de laboratorios múltiples. La región de la mitocondria se utilizó la subunidad 1 de la citocromo c oxidasa como código de barras animal excluido como un marcador potencial, porque es difícil de amplificar en hongos, a menudo incluye intrones grandes y puede ser insuficientemente variable. Tres subunidades del cistrón de ARN ribosómico nuclear fueron en comparación con las regiones de tres genes de codificación proteica representativos (la subunidad más grande de la ARN polimerasa II, la segunda más grande subunidad de la ARN polimerasa II y mantenimiento de minicromosomas proteína). Aunque las regiones de genes que codifican proteínas a menudo tenían un porcentaje más alto de identificación correcta en comparación con ribosomal marcadores, baja amplificación por PCR y éxito de secuenciación eliminados ellos como candidatos para un código de barras universal para hongos. Entre la regiones del cistrón ribosómico, el espaciador transcrito interno (ITS) tiene la mayor probabilidad de identificación exitosa para la gama más amplia de hongos, con la brecha de código de barras más claramente definida entre la variación inter e intraespecífica. El nuclear subunidad ribosómica grande, un marcador filogenético popular en ciertos grupos, tuvieron una resolución de especies superior en algunos grupos taxonómicos, como los primeros linajes divergentes y las levaduras ascomicetas, pero por lo demás era ligeramente inferior al ITS. El ribosomal nuclear La subunidad pequeña tiene una resolución pobre a nivel de especie en los hongos. SU será propuesto formalmente para su adopción como el código de barras fúngico principal marcador al Consorcio para el Código de Barras de la Vida, con la posibilidad de que se desarrollen códigos de barras suplementarios para grupos taxonómicos estrechamente circunscritos., Six DNA regions were evaluated as potential DNA barcodes for Fungi, the second largest kingdom of eukaryotic life, by a multinational, multilaboratory consortium. The region of the mitochondrial cytochrome c oxidase subunit 1 used as the animal barcode was excluded as a potential marker, because it is difficult to amplify in fungi, often includes large introns, and can be insufficiently variable. Three subunits from the nuclear ribosomal RNA cistron were compared together with regions of three representative proteincoding genes (largest subunit of RNA polymerase II, second largest subunit of RNA polymerase II, and minichromosome maintenance protein). Although the protein-coding gene regions often had a higher percent of correct identification compared with ribosomal markers, low PCR amplification and sequencing success eliminated them as candidates for a universal fungal barcode. Among the regions of the ribosomal cistron, the internal transcribed spacer (ITS) region has the highest probability of successful identification for the broadest range of fungi, with the most clearly defined barcode gap between inter- and intraspecific variation. The nuclear ribosomal large subunit, a popular phylogenetic marker in certain groups, had superior species resolution in some taxonomic groups, such as the early diverging lineages and the ascomycete yeasts, but was otherwise slightly inferior to the ITS. The nuclear ribosomal small subunit has poor species-level resolution in fungi. ITS will be formally proposed for adoption as the primary fungal barcode marker to the Consortium for the Barcode of Life, with the possibility that supplementary barcodes may be developed for particular narrowly circumscribed taxonomic groups.

Rhytismatales (Leotiomycetes, Pezizomycotina, Ascomycota) are an order of mostly plant-associated ascomycetes with a global distribution. Well known taxa include the Rhytisma tar spots on Acer spp. and several needle-cast pathogens in genera Lophodermium and Meloderma. Critical studies are lacking at all taxonomic ranks from order to species, and in particular the genus taxonomy in the order has been criticized for being unnatural. We used nuclear LSU and mitochondrial SSU sequences in Bayesian phylogenetic analyses to define a core clade of Rhytismatales sensu stricto. Some of the genera traditionally placed within the Rhytismatales, Ascodichaena, Marthamyces, Mellitiosporium, Potebniamyces, Propolis and Pseudophacidium, are shown to be phylogenetically distinct, all related to various other taxa at present placed in the polyphyletic Helotiales. Within the core clade only Cudonia, Spathularia and Terriera are supported as monophyletic. The large genera Coccomyces, Hypoderma and Lophodermium all are polyphyletic as are a few smaller genera. The traditionally used characters of ascoma and spore shape are shown to be unreliable for the delimitation of monophyletic genera but in some cases can be useful when combined with other characters. In this study we provide 72 new nrLSU and 64 new mtSSU sequences. Together with publicly available sequences data for 103 specimens representing 91 species of Rhytismatales are now available. Despite this taxon sampling intensity is still too low to propose an alternative generic taxonomy.

The diversity and distribution of fungal endophytes in the leaves of four podocarps (Dacrydium cupressinum, Prumnopitys ferruginea, Dacrycarpus dacrydioides, and Podocarpus totara, all Podocarpaceae) and an angiosperm (Kunzea ericoides, Myrtaceae) occurring in close stands were studied. The effects of host species, locality, and season on endophyte assemblages were investigated. Host species was the major factor shaping endophyte assemblages. The spatial separation of sites and seasonal differences played significant but lesser roles. The mycobiota of each host species included both generalist and largely host-specialised fungi. The host-specialists were often observed at low frequencies on some of the other hosts. There was no clear evidence for family-level specialisation across the Podocarpaceae. Of the 17 species found at similar frequencies on several of the podocarp species, 15 were found also on Kunzea. Many of the endophytes isolated appear to represent species of fungi not previously recognised from New Zealand.

We present a 6-gene, 420-species maximum-likelihood phylogeny of Ascomycota, the largest phylum of Fungi. This analysis is the most taxonomically complete to date with species sampled from all 15 currently circumscribed classes. A number of superclass-level nodes that have previously evaded resolution and were unnamed in classifications of the Fungi are resolved for the first time. Based on the 6-gene phylogeny we conducted a phylogenetic informativeness analysis of all 6 genes and a series of ancestral character state reconstructions that focused on morphology of sporocarps, ascus dehiscence, and evolution of nutritional modes and ecologies. A gene-by-gene assessment of phylogenetic informativeness yielded higher levels of informativeness for protein genes (RPB1, RPB2, and TEF1) as compared with the ribosomal genes, which have been the standard bearer in fungal systematics. Our reconstruction of sporocarp characters is consistent with 2 origins for multicellular sexual reproductive structures in Ascomycota, once in the common ancestor of Pezizomycotina and once in the common ancestor of Neolectomycetes. This first report of dual origins of ascomycete sporocarps highlights the complicated nature of assessing homology of morphological traits across Fungi. Furthermore, ancestral reconstruction supports an open sporocarp with an exposed hymenium (apothecium) as the primitive morphology for Pezizomycotina with multiple derivations of the partially (perithecia) or completely enclosed (cleistothecia) sporocarps. Ascus dehiscence is most informative at the class level within Pezizomycotina with most superclass nodes reconstructed equivocally. Character-state reconstructions support a terrestrial, saprobic ecology as ancestral. In contrast to previous studies, these analyses support multiple origins of lichenization events with the loss of lichenization as less frequent and limited to terminal, closely related species.

We constructed a two-locus database, comprising partial translation elongation factor (EF-1 alpha) gene sequences and nearly full-length sequences of the nuclear ribosomal intergenic spacer region (IGS rDNA) for 850 isolates spanning the phylogenetic breadth of the Fusarium oxysporum species complex (FOSC). Of the 850 isolates typed, 101 EF-1 alpha, 203 IGS rDNA, and 256 two-locus sequence types (STs) were differentiated. Analysis of the combined dataset suggests that two-thirds of the STs might be associated with a single host plant. This analysis also revealed that the 26 STs associated with human mycoses were genetically diverse, including several which appear to be nosocomial in origin. A congruence analysis, comparing partial EF-1 alpha and IGS rDNA bootstrap consensus, identified a significant number of conflicting relationships dispersed throughout the bipartitions, suggesting that some of the IGS rDNA sequences may be non-orthologous. We also evaluated enniatin, fumonisin and moniliformin mycotoxin production in vitro within a phylogenetic framework

The B trichothecene toxin-producing clade (B clade) of Fusarium includes the etiological agents of Fusarium head blight, crown rot of wheat and barley and stem and ear rot of maize. B clade isolates also have been recovered from several wild and cultivated grasses, including Dactylis glomerata (orchard grass or cock's foot), one of the world's most important forage grasses. Two isolates from the latter host are formally described here as F. dactylidis. Phenotypically F. dactylidis most closely resembles F. ussurianum from the Russian Far East. Both species produce symmetrical sporodochial conidia that are similar in size and curved toward both ends. However, conidia of F. ussurianum typically end in a narrow apical beak while the apical cell of F. dactylidis is acute. Fusarium dactylidis produced nivalenol mycotoxin in planta as well as low but detectable amounts of the estrogenic mycotoxin zearalenone in vitro. Results of a pathogenicity test revealed that F. dactylidis induced mild head blight on wheat.

Phylogenetic relationships of one of the largest nonlichen-forming ascomycetous groups, the Leotiomycetes, were inferred from genes encoding three rDNA regions (SSU+LSU+5.8S rDNA). A dataset was prepared with rDNA sequences data from 108 isolates, among which we sampled 85 taxa representing four orders and 16 families in the Leotiomycetes. Equally weighted parsimony and Bayesian analyses were performed. Bootstrap proportion and Bayesian posterior probability under the GTR+gamma+iota model were estimated along the branches. Based on our results the Leotiomycetes is relatively well defined as a class and it includes the Cyttariales, Erysiphales, Helotiales, Rhytismatales and two families of uncertain position, Myxotrichaceae and Pseudeurotiaceae. The placements of the Thelebolales and Ascocorticiaceae are not examined and are accepted as tentative in the Leotiomycetes. Our results agree with previous studies to remove the Geoglossaceae, including Geoglossum, Trichoglossum and Sarcoleotia, from the Leotiomycetes. Positions of the Erysiphales and Rhytismatales in the Leotiomycetes are confirmed. The Helotiales and Myxotrichaceae are paraphyletic. Close relationships are supported strongly among the Hemiphacidiaceae, Rutstroemiaceae and Sclerotiniaceae, among Loramycetaceae, the northern hemisphere Vibrisseaceae, the Dark Septate Endophyte fungus Phialocephala fortinii and Mollisia cinerea, and between species of Bulgaria and Phadidiopycnis. Sequence data of rDNA regions are not adequate to resolve the relationships among major groups of the Leotiomycetes.

The highly divergent characters of morphology, ecology, and biology in the Helotiales make it one of the most problematic groups in traditional classification and molecular phylogeny. Sequences of three rDNA regions, SSU, LSU, and 5.8S rDNA, were generated for 50 helotialean fungi, representing 11 out of 13 families in the current classification. Data sets with different compositions were assembled, and parsimony and Bayesian analyses were performed. The phylogenetic distribution of lifestyle and ecological factors was assessed. Plant endophytism is distributed across multiple clades in the Leotiomycetes. Our results suggest that (1) the inclusion of LSU rDNA and a wider taxon sampling greatly improves resolution of the Helotiales phylogeny, however, the usefulness of rDNA in resolving the deep relationships within the Leotiomycetes is limited; (2) a new class Geoglossomycetes, including Geoglossum, Trichoglossum, and Sarcoleotia, is the basal lineage of the Leotiomyceta; (3) the Leotiomycetes, including the Helotiales, Erysiphales, Cyttariales, Rhytismatales, and Myxotrichaceae, is monophyletic; and (4) nine clades can be recognized within the Helotiales.

A diverse collection of isolates identified as Colletotrichum acutatum, including a range of fruit-rot and foliar pathogens, was examined for mtDNA RFLPs and RFLPs and sequence variation of a 900-bp intron of the glutamine synthetase (GS) gene and a 200-bp intron of the glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene. RFLPs of mtDNA, RFLPs of the 900-bp GS intron and sequence analysis of each intron identified the same seven distinct molecular groups, or clades, within C. acutatum sensu lato. Sequence analysis produced highly concordant tree topologies with definitive phylogenetic relationships within and between the clades. The clades might represent phylogenetically distinct species within C. acutatum sensu lato. Mating tests also were conducted to assess sexual compatibility with tester isolates known to outcross to form the teleomorph Glomerella acutata. Mating compatibility was identified within one clade, C, and between two phylogenetically distinct clades, C and J4. The C clade represented isolates from a wide range of hosts and geographic origins. J4 clade contained isolates from Australia or New Zealand recovered from fruit rot and pine seedlings with terminal crook disease. That isolates in two phylogenetically distinct clades were capable of mating suggests that genetic isolation occurred before reproductive isolation. No other isolates were sexually compatible with the mating testers, which also were in groups C and J4. Certain clades identified by mtDNA and intron analysis (D1, J3 and J6) appeared to represent relatively host-limited populations. Other clades (C1, F1 and J4) contained isolates from a wide range of hosts. Isolates described as C. acutatum f. sp. pineum were clearly polyphyletic.

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Reannotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi., B.R. and C.L.S. acknowledge support from the Intramural Research Program of the National Institutes of Health, National Library of Medicine

The genetic relationships between several morphological groups recognized within Colletotrichum acutatum sensu lato were investigated using RAPD analysis and vegetative compatibility analysis. Isolates were examined from fruit rots originating in New Zealand and Australia, from Lupinus spp. in New Zealand, Canada, France and the U.K., and from Pinus radiata in New Zealand and Australia. The genetic distinctness of the groups recognized morphologically is confirmed. Two genetically distinct groups of C. acutatum-like pathogens are recognized from lupin, one comprising isolates from New Zealand and the U.K., the other isolates from Canada and France. C. acutatum f. sp. pineum isolates from New Zealand and Australia form two genetically distinct groups.

The diversity and frequency of leaf endophytic fungi occurring in naturally regenerating and planted stands of manuka were investigated at several sites. A restricted and characteristic range of species was isolated from all sites, although marked differences were noted between natural and planted stands. In natural stands a Phyllosticta species was the single dominant endophyte. This fungus appears to be host species specific, as it was not isolated from the closely related kanuka (Kunzea ericoides) in mixed, even-aged stands of manuka and kanuka. The frequency of the Phyllosticta sp. fluctuated greatly between individual trees across single stands. No obvious environmental or other gradients were recognized which could explain this patchy distribution. The Phyllosticta sp., along with the two next most common species in natural stands, Diploceras leptospermi and an unidentified coelomycete, were absent from most of the planted stands sampled, and at very low frequencies when present. Although several of the planted stands were close to natural stands and had been established for several years, the difference between them has been maintained. The most common endophytes in planted stands were Botryosphaeria and Alternaria spp. These species, found at low levels in natural stands, are likely to be ‘opportunistic’ endophytes. They are probably also characteristic of the epiphytic mycota, invading only the living leaves of plants under stress. The difference in leaf endophyte communities between planted and natural stands of manuka may be an indication of wider differences in the microbial communities of the respective stands, and could prove significant for site restoration projects. Such projects using nursery-raised manuka plants as a pioneer species may be establishing stands lacking much of the natural diversity of manuka communities for extended periods following establishment.

Isolates of Colletotrichum associated with fruit rots in New Zealand were used to test the applicability of morphological, cultural, and rDNA sequence data to clarify relationships within this genu...

We constructed several multilocus DNA sequence datasets to assess the phylogenetic diversity of insecticolous fusaria, especially focusing on those housed in the Agricultural Research Service Collection of Entomopathogenic Fungi (ARSEF), and to facilitate molecular identifications of unknowns via the FUSARIUM-ID and Fusarium MLST online databases and analysis packages. Analyses of a 190-taxon two-locus dataset, which included 159 isolates from insects, indicated that: 1) insect-associated fusaria were nested within 10 species complexes spanning the phylogenetic breadth of Fusarium, 2) novel, putatively unnamed insecticolous species were nested within 8/10 species complexes, and 3) Latin binomials could be applied with confidence to only 18/58 phylogenetically distinct fusaria associated with pest insects. Phylogenetic analyses of an 82-taxon three-locus dataset nearly fully resolved evolutionary relationships among the 10 clades containing insecticolous fusaria. Multilocus typing of isolates within four species complexes identified surprisingly high genetic diversity in that 63/65 of the fusaria typed represented newly discovered haplotypes. The DNA sequence data, together with corrected ABI sequence chromatograms and alignments, have been uploaded to the following web-accessible sites dedicated to identifying fusaria: FUSARIUM-ID (http://isolate.fusariumdb.org) at The Pennsylvania State University's Department of Plant Pathology and Fusarium MLST (http://www.cbs.knaw.nl/fusarium) at the Centraalbureau voor Schimmelcultures (CBS-KNAW) Fungal Biodiversity Center.

Single-ascospore isolates from perithecia belonging to a species of Gibberella found on diseased gorse ( Ulex europaeus ) plants produced an anamorph in culture identified as Fusarium tumidum . A teleomorph has not previously been reported for F. tumidum and it is described as Gibberella tumida sp. nov. ascospore size distinguishing it from all previously described species.

Lophodermium comprises ascomycetous fungi that are both needle-cast pathogens and asymptomatic endophytes on a diversity of plant hosts. It is distinguished from other genera in the family Rhytismataceae by its filiform ascospores and ascocarps that open by a longitudinal slit. Nucleotide sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA were used to infer phylogenetic relationships within Lophodermium. Twenty-nine sequences from approximately 11 species of Lophodermium were analyzed together with eight sequences from isolates thought to represent six other genera of Rhytismataceae: Elytroderma, Lirula, Meloderma, Terriera, Tryblidiopsis and Colpoma. Two putative Meloderma desmazieresii isolates occurred within the Lophodermium clade but separate from one another, one grouped with L. indianum and the other with L. nitens. An isolate of Elytroderma deformans also occurred within the Lophodermium clade but on a solitary branch. The occurrence of these genera within the Lophodermium clade might be due to problems in generic concepts in Rhytismataceae, such as emphasis on spore morphology to delimit genera, to difficulty of isolating Rhytismataceae needle pathogens from material that also is colonized by Lophodermium or to a combination of both factors. We also evaluated the congruence of host distribution and several morphological characters on the ITS phylogeny. Lophodermium species from pine hosts formed a monophyletic sister group to Lophodermium species from more distant hosts from the southern hemisphere, but not to L. piceae from Picea. The ITS topology indicated that Lophodermium does not show strict cospeciation with pines at deeper branches, although several closely related isolates have closely related hosts. Pathogenic species occupy derived positions in the pine clade, suggesting that pathogenicity has evolved from endophytism. A new combination is proposed, Terriera minor (Tehon) P.R. Johnst.

A new genus, Claviradulomyces, morphologically typical of Odontotremataceae, is proposed to accommodate the new species Claviradulomyces dabeicola. The genus is characterised by its ornamented periphysoids, and long-cylindric, sigmoid, acuminate ascospores. Genetically, C. dabeicola falls within the Ostropales, but the Odontotremataceae have been very poorly sampled genetically and their phylogenetic relationship within the order is unclear. C. dabeicola has been collected consistently and exclusively on the rainforest tree Erythroxylum mannii in the Cote d'Ivoire and Ghana, where it is associated with bark abnormalities on branches and stems of living trees.

Asci of most members of the Coryneliaceae were examined. All species of Corynelia, Coryneliospora Fitzpatrickiella, Lagenulopsis , and Tripospora , and representative species of Caliciopsis and Coryneliopsis were studied by light microscopy. Two species of Corynelia were also examined using transmission electron microscopy. Except in Coryneliopsis , all had asci with more than one functional wall layer with the outer layer breaking during ascus elongation, long before ascospores are ready to be released. It is concluded that the order Coryneliales, containing the single family Coryneliaceae, should be retained, and that Coryneliopsis should probably be removed from the family.