Generic boundaries in the Ophiostomatales reconsidered and revised

Abstract

The Ophiostomatales was erected in 1980. Since that time, several of the genera have been redefined and others have been described. There are currently 14 accepted genera in the Order. They include species that are the causal agents of plant and human diseases and common associates of insects such as bark beetles. Well known examples include the Dutch elm disease fungi and the causal agents of sporotrichosis in humans and animals. The taxonomy of the Ophiostomatales was confused for many years, mainly due to the convergent evolution of morphological characters used to delimit unrelated fungal taxa. The emergence of DNA-based methods has resolved much of this confusion. However, the delineation of some genera and the placement of various species and smaller lineages remains inconclusive. In this study we reconsidered the generic boundaries within the Ophiostomatales. A phylogenomic framework constructed from genome-wide sequence data for 31 species representing the major genera in the Order was used as a guide to delineate genera. This framework also informed our choice of the best markers from the currently most commonly used gene regions for taxonomic studies of these fungi. DNA was amplified and sequenced for more than 200 species, representing all lineages in the Order. We constructed phylogenetic trees based on the different gene regions and assembled a concatenated data set utilising a suite of phylogenetic analyses. The results supported and confirmed the delineation of nine of the 14 currently accepted genera, i.e. Aureovirgo, Ceratocystiopsis, Esteya, Fragosphaeria, Graphilbum, Hawksworthiomyces, Ophiostoma, Raffaelea and Sporothrix. The two most recently described genera, Chrysosphaeria and Intubia, were not included in the multi-locus analyses. This was due to their high sequence divergence, which was shown to result in ambiguous taxonomic placement, even though the results of phylogenomic analysis supported their inclusion in the Ophiostomatales. In addition to the currently accepted genera in the Ophiostomatales, well-supported lineages emerged that were distinct from those genera. These are described as novel genera. Two lineages included the type species of Grosmannia and Dryadomyces and these genera are thus reinstated and their circumscriptions redefined. The descriptions of all genera in the Ophiostomatales were standardised and refined where this was required and 39 new combinations have been provided for species in the newly emerging genera and one new combination has been provided for Sporothrix. The placement of Afroraffaelea could not be confirmed using the available data and the genus has been treated as incertae sedis in the Ophiostomatales. Paleoambrosia was not included in this study, due to the absence of living material available for this monotypic fossil genus. Overall, this study has provided the most comprehensive and robust phylogenies currently possible for the Ophiostomatales. It has also clarified several unresolved One Fungus-One Name nomenclatural issues relevant to the Order.

Taxonomic novelties: New genera: Harringtonia Z.W. de Beer & M. Procter, Heinzbutinia Z.W. de Beer & M. Procter, Jamesreidia Z.W. de Beer & M. Procter, Masuyamyces Z.W. de Beer & M. Procter. New species: Masuyamyces massonianae M. Procter & Z.W. de Beer. New combinations: Dryadomyces montetyi (M. Morelet) M. Procter & Z.W. de Beer, Dryadomyces quercivorus (Kubono & Shin. Ito) M. Procter & Z.W. de Beer, Dryadomyces quercus-mongolicae (K.H. Kim et al.) M. Procter & Z.W. de Beer, Dryadomyces sulphureus (L.R. Batra) M. Procter & Z.W. de Beer, Graphilbum pusillum (Masuya) M. Procter & Z.W. de Beer, Grosmannia abieticolens (K. Jacobs & M.J. Wingf.) M. Procter & Z.W. de Beer, Grosmannia altior (Paciura et al.) M. Procter & Z.W. de Beer, Grosmannia betulae (Jankowiak et al.) M. Procter & Z.W. de Beer, Grosmannia curviconidia (Paciura et al.) M. Procter & Z.W. de Beer, Grosmannia euphyes (K. Jacobs & M.J. Wingf.) M. Procter & Z.W. de Beer, Grosmannia fenglinhensis (R. Chang et al.) M. Procter & Z.W. de Beer, Grosmannia gestamen (de Errasti & Z.W. de Beer) M. Procter & Z.W. de Beer, Grosmannia innermongolica (X.W. Liu et al.) M. Procter & Z.W. de Beer, Grosmannia pistaciae (Paciura et al.) M. Procter & Z.W. de Beer, Grosmannia pruni (Masuya & M.J. Wingf.) M. Procter & Z.W. de Beer, Grosmannia taigensis (Linnak. et al.) M. Procter & Z.W. de Beer, Grosmannia trypodendri (Jankowiak et al.) M. Procter & Z.W. de Beer, Harringtonia aguacate (D.R. Simmons et al.) M. Procter & Z.W. de Beer, Harringtonia brunnea (L.R. Batra) M. Procter & Z.W. de Beer, Harringtonia lauricola (T.C. Harr. et al.) Z.W. de Beer & M. Procter, Heinzbutinia grandicarpa (Kowalski & Butin) Z.W. de Beer & M. Procter, Heinzbutinia microspora (Arx) M. Procter & Z.W. de Beer, Heinzbutinia solheimii (B. Strzałka & Jankowiak) Z.W. de Beer & M. Procter, Jamesreidia coronata (Olchow. & J. Reid) M. Procter & Z.W. de Beer, Jamesreidia nigricarpa (R.W. Davidson) M. Procter & Z.W. de Beer, Jamesreidia rostrocoronata (R.W. Davidson & Eslyn) M. Procter & Z.W. de Beer, Jamesreidia tenella (R.W. Davidson) Z.W. de Beer & M. Procter, Leptographium cainii (Olchow. & J. Reid) M. Procter & Z.W. de Beer, Leptographium europioides (E.F. Wright & Cain) M. Procter & Z.W. de Beer, Leptographium galeiforme (B.K. Bakshi) M. Procter & Z.W. de Beer, Leptographium pseudoeurophioides (Olchow. & J. Reid) M. Procter & Z.W. de Beer, Leptographium radiaticola (J.J. Kim et al.) M. Procter & Z.W. de Beer, Masuyamyces acarorum (R. Chang & Z.W. de Beer) M. Procter & Z.W. de Beer, Masuyamyces ambrosius (B.K. Bakshi) M. Procter & Z.W. de Beer, Masuyamyces botuliformis (Masuya) Z.W. de Beer & M. Procter, Masuyamyces jilinensis (R. Chang et al.) M. Procter & Z.W. de Beer, Masuyamyces lotiformis (Z. Wang & Q. Lu) M. Procter & Z.W. de Beer, Masuyamyces pallidulus (Linnak. et al.) M. Procter & Z.W. de Beer, Masuyamyces saponiodorus (Linnak. et al.) M. Procter & Z.W. de Beer, Sporothrix longicollis (Massee & E.S. Salmon) M. Procter & Z.W. de Beer.

Citation: de Beer W, Procter M, Wingfield MJ, Marincowitz S, Duong TA (2022). Generic boundaries in the Ophiostomatales reconsidered and revised. Studies in Mycology 101: 57–120. doi: 10.3114/sim.2022.101.02

INTRODUCTION

The Ophiostomatales (Sordariomycetidae, Ascomycota), was described by Benny & Kimbrough (1980) accommodating the single family Ophiostomataceae. The Order includes species that are the causal agents of plant and human diseases as well as common associates of wood infesting insects such as bark beetles (Wingfield et al. 1993, Seifert et al. 2013). Well known examples are the Dutch elm disease fungi and the causal agents of sporotrichosis in humans and animals (Figs 1, 2). As described, this Order originally accommodated four genera, Ophiostoma, Ceratocystiopsis, Sphaeronaemella and Ceratocystis. These authors considered Ceratocystis distinct from Ophiostoma based on cell wall constituents and conidiogenesis (Weijman & De Hoog 1975, Seifert et al. 2013). Upadhyay (1981) treated Ophiostoma as a synonym of Ceratocystis based on morphological similarities such as their long-necked ascocarps that produce sheathed ascospores in sticky droplets to facilitate arthropod dispersal (Upadhyay 1981, Malloch & Blackwell 1993). In the 1990’s, DNA sequence data confirmed that Ophiostoma and Ceratocystis resided in two distinct Orders of the fungi (Hausner et al. 1993c, Spatafora & Blackwell 1994). Ceratocystis has subsequently been shown to represent several morphologically and ecologically distinct genera in the Ceratocystidaceae (Microascales) (De Beer et al. 2013a, 2014, Nel et al. 2018, Mayers et al. 2015, 2020).

Fig. 1.

Ecological niches in which genera and species residing in the Ophiostomatales are found. A. Ulmus americana street trees dying as a result of Dutch Elm Disease (photo: D.W. French). B. Symptoms of infection by the human pathogen Sporothix schenckii (photo: Prof. Dr Flávio de Queiroz Telles Filho, Federal University of Paraná, Brazil). C. Signage in Yellowstone National Park emphasising the important role that bark beetles (and by extension their fungal symbionts) play in the ecology of conifer ecosystems. D. Blue stain in conifer timber caused by numerous species of Ophiostomatoid fungi. E. Hylobius rhizophagus (root collar weevil) squashed onto the surface of agar medium containing cycloheximide selective for many genera and species of Ophiostomatales and in this case Leptographium procerum. F. Infructescences of a Protea species in which numerous species of Ophiostomatoid fungi can be found. G. Douglas fir (Pseudotsuga menzesii) trees dying as a result of black stain root disease caused by Leptographium wageneri var. pseudotsugae (Photo: F.W. Cobb). H. Pinus resinosa trees dying as a result of mass infestation by Ips pini and associated Ophiostoma minus.

Fig. 2.

Ecological niches of Ophiostomatoid fungi and micrographs providing examples of structures typical of these fungi. A. Transverse stellate gallery systems of Ips schmutzenhoferi in the bark of a Pinus spinulosa tree, showing blue-stain around nuptial chambers and female galleries. B. Section through a Eucalyptus stem infested by the ambrosia beetle Megaplatypus mutatus showing tunnels in which species of Ophiostomatales occur. C, D. Ascomata of Ophiostoma ulmi (C) (photo: D.W. French) and O. pilliferum (D) (photo: Z.W. de Beer) with sticky ascospores masses at their apices, illustrating the manner by which these fungi easily attach to the insects that carry them. E. Conidiophores of Leptographium procerum, illustrating asexual structures well suited to being vectored by insects. F. Conidiogenous cells of a Sporothrix sp. G. Typical single-celled conidia found in most species of Ophiostomatales. H. Many species of Ophiostomatales have ascospores with sheaths such as these pillow-shaped spores in Ophiostoma ips.

The Ophiostomatales as it is currently defined accommodates a single family, the Ophiostomataceae (De Beer et al. 2013a), which was initially described in 1932 (Nannfeldt 1932). At the time, it included Ophiostoma, with Endoconidiophora and Ceratocystis as synonyms (Melin & Nannfeldt 1934). The family was treated in various Orders prior to 1980 (De Beer et al. 2013a). Apparently unaware of Benny & Kimbrough’s (1980) study, Upadhyay (1981) re-defined the Ophiostomataceae with Ceratocystis as type genus, and Ophiostoma, Sphaeronaemella, Grosmannia and Europhium as synonyms, and with Ceratocystiopsis as a distinct new genus. However, the DNA-based distinction between Ceratocystis and Ophiostoma led to their inevitable separation and emended definitions of the Ceratocystidaceae and Ophiostomataceae (Wingfield et al. 1993, Réblová et al. 2011, De Beer et al. 2013a, 2016a).

The first attempt to resolve generic boundaries within the Ophiostomatales subsequent to its separation from the Ceratocystidaceae was made by Zipfel et al. (2006). Based on phylogenies constructed from ribosomal DNA and β-tubulin sequences and including 55 taxa, they recognised Ophiostoma, Ceratocystiopsis and Grosmannia as distinct sexual genera. Following the dual nomenclature system at the time, asexual Sporothrix and Leptographium species retained their names in these genera, although they respectively grouped in Ophiostoma and Grosmannia.

After 20 years of DNA-based taxonomy and in the wake of the abandonment of a dual nomenclature for the fungi (Hawksworth et al. 2011, Hawksworth 2012), De Beer & Wingfield (2013) revised the Ophiostomatales, considering all published ribosomal large subunit (LSU) and internal transcribed spacer (ITS) sequences, including 266 species. They redefined Ophiostoma sensu stricto, Raffaelea s.s., Ceratocystiopsis, Fragosphaeria and Graphilbum, and recognised 18 species complexes within the various genera of the Ophiostomatales. However, they concluded that the rDNA-based phylogenies were not sufficiently robust to resolve the generic status of the Sporothrix schenckii-Ophiostoma stenoceras species complex, or that of various lineages within what they defined as Leptographium sensu lato. Their phylogenies also suggested that ambrosial species previously treated as Raffaelea did not form a monophyletic group.

After the revision of the Ophiostomatales by De Beer & Wingfield (2013), the majority of studies focused on new species descriptions (Romón et al. 2014a, b, Musvuugwa et al. 2015, 2016, De Errasti et al. 2016, Simmons et al. 2016, Chang et al. 2017, Marincowitz et al. 2017, 2020, etc.) and on resolving issues within species complexes (Ando et al. 2016, Linnakoski et al. 2016, Jankowiak et al. 2017, Yin et al. 2019, 2020, etc.).

De Beer et al. (2016a) reconsidered the status of the S. schenckii-O. stenoceras complex, providing sequences for four gene regions of 65 species with sporothrix-like asexual morphs. They concluded that Sporothrix represented a distinct genus including 51 species and incorporated the characters of the sexual morphs of many of the species, previously treated as Ophiostoma, in the emended definition of Sporothrix. A lineage including some of the remaining sporothrix-like species that did not form part of the newly defined genus were provided with the new genus name, Hawksworthiomyces, in a subsequent paper (De Beer et al. 2016b). In addition, Van der Linde et al. (2016) and Bateman et al. (2017) described Aureovirgo and Afroraffaelea respectively as novel, monotypic genera in the Order. In 2018, a fungus was discovered preserved in amber alongside an ambrosia beetle, leading to the description of Paleoambrosia. The genus was treated in the Ophiostomatales based on morphological characters resembling Raffaelea species (Poinar & Vega 2018). Most recently, Nel et al. (2021) described two new genera, Chrysosphaeria and Intubia, from the abandoned combs of fungus-growing termites (Termitomyces) in South Africa.

In this study, we reconsidered and redefined the unresolved boundaries of genera including Leptographium, Raffaelea and some smaller lineages in the Ophiostomatales. To achieve this goal, we selected four gene regions based on a phylogenomic framework constructed from genome-wide sequence data for representative ophiostomatalean species. Sequence data for these four gene regions were then generated for as many species in the Order as possible, and phylogenetic analyses were conducted.

MATERIALS AND METHODS

Fungal isolates and DNA extraction

Fungal cultures used in this study were obtained from the Culture Collection (CMW) of the Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, South Africa, and the Westerdijk Fungal Biodiversity Institute (CBS), Utrecht, the Netherlands. Isolates were grown on 2 % malt extract agar (MEA: 20 g malt extract, 20 g agar, 1 L dH₂O) at room temperature; initially with streptomycin (0.4 g/L, Sigma-Aldrich, Kempton Park, South Africa) and cycloheximide (0.5 g/L, Sigma-Aldrich, Kempton Park, South Africa) supplemented in the media, then sub-cultured to MEA, and maintained at 4 °C after optimal growth had occurred. DNA was extracted following the protocol described by Duong et al. (2012).

Phylogenomic analyses

Available genome sequences for 31 species representing 11 of 14 currently recognised genera (excluding Afroraffaelea, Aureovirgo and Paleoambrosia) in the Ophiostomatales (Supplementary Table S1) were used to construct a phylogenomic tree. Two species of Diaporthales (Cryphonectria parasitica and Diaporthe ampelina), two species of Magnaporthales (Magnaporthe grisea and Magnaporthe poae), and one species of Togniniales (Phaeoacremonium minimum) were included in the analyses as outgroup taxa. Genome assemblies for all species were subjected to BUSCO v. 4.0.5 (Seppey et al. 2019) runs using the Sordariomycetes_odb10 dataset to obtain BUSCO genes. The amino acid sequences were extracted from BUSCO results and datasets were compiled for each of the BUSCO orthologous groups. All BUSCO orthologous groups with duplicated BUSCO genes were excluded from the analysis. PRANK (Löytynoja 2014) was used to align the datasets with default parameters.

Trimal v. 1.4 (Capella-Gutiérrez et al. 2009) was used for trimming of the alignments with “-resoverlap 0.8 -seqoverlap 75” parameters. Only datasets with lengths equal or larger than 100 aa after trimming step were retained for further analysis. Permutation Tail Probability (PTP) tests were conducted in PAUP v. 4.0a (Swofford 2003) to identify and remove datasets having no phylogenetic signal as well as those with less than 50 parsimony-informative characters. Individual gene trees for each BUSCO orthologous group were constructed using IQ-TREE v. 2 with an optimal substitution model automatically determined and 1 000 ultrafast bootstraps (Hoang et al. 2018, Minh et al. 2020). TreeShrink v. 1.3.7 (Mai & Mirarab 2018) was used to remove outliers (taxa with abnormal branch length) from all trees with default parameters. Newick utilities (Junier & Zdobnov 2010) was used to collapse branches with less than 10 % bootstrap support. A species tree was then constructed from the final set of trees under the multi-species coalescent model using ASTRAL v. 5.7.7 (Mirarab et al. 2014). Branch length of the species tree was optimized with RAxML v. 8.2.11 (Stamatakis 2014) using only BUSCO orthogroups that have all 36 taxa present and without any outliner taxon as identified with TreeShrink analysis.

Selection of gene regions for phylogenetic analyses

Sequences for eight gene regions commonly used in phylogenetic studies of the fungi were extracted from 31 draft genome sequences for species in the Ophiostomatales, as well as from those used as outgroups in the phylogenomic analysis (Supplementary Table S1). These included β-tubulin (β-tub), translation elongation factor 1 alpha (TEF-1α), internal transcribed spacer region (ITS), ribosomal large subunit (LSU), mini chromosome maintenance protein complex 7 (MCM7), DNA-directed RNA polymerase II second largest subunit (RPBII), DNA-directed RNA polymerase II largest subunit (RPBI) and ribosomal small subunit (SSU). A phylogenetic tree was constructed for each of these datasets using IQ-TREE v. 2 as indicated above. Based on the level of congruency between individual gene trees and the phylogenomic tree as well as the phylogenetic signal of these gene regions, the LSU, ITS, TEF-1α and RPBII gene regions were selected as markers to delineate genera in the Ophiostomatales. These regions included the primary (ITS) and secondary (TEF-1α) barcodes for fungi (Schoch et al. 2012, Stielow et al. 2015).

Primer selection

Existing primers were used to amplify and sequence the ITS (ITS1F: Gardes & Bruns 1993, ITS4: White et al. 1990), TEF-1α (EF2F: Marincowitz et al. 2015, EF2R: Jacobs et al. 2004) and LSU (LR5, LROR: Vilgalys & Hester 1990) gene regions. Since previously available primers for RPBII did not consistently amplify the targeted region in most of the isolates investigated, we designed new primers for this gene region based on the available genome sequences: Oph-RPB2F1 (5’ - GAYGAYCGIGAYCAYTTYGG - 3’), Oph-RPB2F2 (5’ - TICTGGCIAARCTNTTCCG - 3’) and Oph-RPB2R1 (5’ - CCCATRGCYTGYTTRCCCAT - 3’). A combination of Oph-RPBF1 and Oph-RPBR1 was used in most instances, while Oph-RPBF2 and Oph-RPBR1 were used in cases where the former combination was not successful.

PCR

For the ITS, TEF-1α and LSU regions, FastStart Taq DNA Polymerase (Roche, Germany) was used. For the RPBII gene region the Platinum^® Multiplex PCR Master Mix (Applied Biosystems, Foster City, California) was used. The ITS, LSU and TEF-1α gene regions were amplified following the protocol described by Duong et al. (2012). For the RPBII gene region, the protocol provided with the Platinum^® Multiplex PCR Master Mix was used but amended as follows: the PCR mixture was made up to a final volume of 12.5 μL, primers were added to a concentration of 1 μM each, and PCR was carried out with 40 cycles of denaturing at 94 °C for 30 s, annealing at 58 °C for 30 s, and elongation at 72 °C for 1 min.

Agarose gel electrophoresis (1 % agarose) was performed on all PCR products to confirm the success of amplification. PCR products were treated with ExoSAP (a mixture of exonuclease I and alkaline phosphatase; one unit of each enzyme was used for approximately 20 μL of PCR product). The mixture was then subjected to two incubation steps at 37 °C for 15 min (for enzymatic action) and 80 °C for 15 min (to deactivate the enzymes). The treated products were stored at 4 °C until PCR sequencing was carried out.

DNA sequencing

Sanger sequencing was performed for all ExoSAP treated PCR products. The PCR sequencing setup reaction (12 μL) consisted of 6.4 μL dH₂O, 2.1 μL 5× sequencing buffer, 0.5 μL BigDye v 3.1, 1 μL of the forward or reverse primer (10 mM), and 2 μL ExoSAP treated PCR product. The reaction was performed under the following conditions: 25 cycles of a denaturing step at 96 °C for 10 s, an annealing step at 55 °C for 5 s, and an elongation step at 60 °C for 4 min. The products were maintained at 4 °C until being used for precipitation.

PCR sequencing products were precipitated using the ethanol/NaOAc precipitation method. For each of the PCR sequencing products (total volume of 12 μL) 8 μL dH₂O, 2 μL NaOAc (3 M, pH 5.2) and 50 μL absolute ethanol (EtOH) was added. The tubes were incubated on ice for 10 min, then centrifuged at 13 400 rpm for 30 min at room temperature. After centrifugation, the supernatant was removed, and the pellet washed twice with 150 μL of 70 % EtOH and centrifuged for 10 min at 13 400 rpm at room temperature. After the final wash, the supernatant was removed, and the pellet was air-dried for approximately 15 min. The samples were kept at -20 °C until they could be analysed. The fragment separations were performed using an ABI PRISM^® 3100 Genetic Analyzer (Applied Biosystems). Consensus sequences were derived from sequences obtained with forward and reverse primers. All sequences generated in this study have been submitted to GenBank, and those from ex-type isolates will be included in the RefSeq Targeted Loci (RTL) database in GenBank (Schoch et al. 2014).

Phylogenetic analyses

For species with available genome sequences (Supplementary Table S1), gene region data were extracted from assembled genome sequences. Sequences were downloaded from GenBank for newly described species for which cultures were not available during the study period, as well as for species for which our sequence data were incomplete. Datasets were aligned using the online version of MAFFT v. 7 (Katoh & Stanley 2013) with default parameters. Alignments were refined with an online version of Gblocks v. 0.91b (Castresana 2000) using default parameters – i.e. no alternative options for more or less stringent selection were selected. Datasets for the various gene regions obtained from Gblocks were concatenated using FASconCAT-G (Kück & Meusemann 2010). Partitionfinder v. 2.1.1 (Lanfear et al. 2017) was used to determine best substitution models for the combined dataset.

Maximum Likelihood (ML) analyses using RAxML (Stamatakis 2014) were performed separately for all gene regions and for the concatenated dataset with raxmlGUI v. 1.3 (Silvestro & Michalak 2012) using the GTR+G+I substitution model and 1 000 thorough bootstrap replicates. Bayesian analysis was conducted using PhyloBayes-MPI v. 1.8 (Lartillot et al. 2013); two chains were run in parallel under the CAT-GTR model. The program bpcomp was used to assess the convergence in tree space. Runs were terminated when the maxdiff value obtained between two chains reached 0.1 or lower.

Morphology

Of the 11 ascospore morphotypes defined by De Beer & Wingfield (2013) for the Ophiostomatales, three (Type A, B, E) were used to describe ascospore types in the present study (Fig. 3). Type A ascospores are those described as allantoid, bean-shaped, crescent to sickle-shaped, clavate to ovate, curved, cylindrical and slightly curved, orange section, lunate or reniform. Type B ascospores are those that are bacilliform, elongate filiform or narrow clavate. Type E ascospores are those with sheaths and are box-shaped, cylindrical, oblong, pillow-shaped, rectangular or rod-shaped. For the asexual morphs, five conidiophore types, hyalorhinocladiella-like, leptographium-like, pesotum-like, raffaelea-like and sporothrix-like, were used as descriptors where applicable (Fig. 3). Three shades of grey were applied in the figures to depict the colours of structures. Thus, hyaline to subhyaline structures were shaded in pale grey. Brown to dark brown structures were a medium-tone grey and fuscous black to black structures were presented in dark grey.

Fig. 3.

Conidiophore and ascospore types mentioned in the paper. A–E. Conidiophore types. F–H. Ascospore types sensu De Beer & Wingfield (2013). A, B, D, F–H. Adapted from illustrations in De Beer & Wingfield (2013). Shades of grey depict colours of various structures ranging from hyaline to dematiaceous.

RESULTS

The phylogenomic tree constructed from the genome-wide sequence data for 31 ophiostomatalean species (Fig. 4) showed a similar topology to those obtained in previous studies (Nel et al. 2021, Vanderpool et al. 2018). The placement of Graphilbum fragrans was, however, different from that suggested by Vanderpool et al. (2018). This inconsistent placement for Gra. fragrans was also observed by Nel et al. (2021) where different phylogenomic approaches were applied. Species of Leptographium s.l. grouped in two distinct lineages; one of these accommodated the Grosmannia penicillata complex and the other the L. lundbergii, L. procerum and L. galeiforme complexes. Species of Raffaelea s.l. resolved in three distinct clades, which is consistent with the findings in two previous studies (Nel et al. 2021, Vanderpool et al. 2018). Otherwise, all remaining species and genera included in the analysis resided in the clades consistent with their current recognition as separate genera in the Ophiostomatales.

Fig. 4.

Phylogenomic tree obtained from supertree analysis with ASTRAL using gene trees constructed from 3 548 BUSCO genes (identified using the sordariomycetes_odb10 dataset; BUSCO v. 4.0.5). All 31 species in the Ophiostomatales for which genome sequence currently available were included in the analysis. Cryphonectria parasitica, Diaporthe ampelina, Magnaporthe grisea, Magnaporthe poae and Phaeoacremonium minimum were included as outgroup taxa. Gene concordance factors (gCF) and site concordance factors (sCF), which indicate the percentage of genes and sites that support a parcular nodes respectively, were determined using IQ-TREE2 are presented at nodes as gCF/sCF. Species names presented in double quotes denote old names which have been changed to their new respective genera subsequent to this study.

The maximum likelihood tree (Fig. 5) resulting from analyses of the concatenated dataset (LSU, ITS, TEF-1α and RPBII) for 264 isolates representing 249 species revealed 24 distinct lineages. Posterior probability values generated from Bayesian Inference analysis are indicated at the genus-level nodes (Fig. 5). Although the topologies of the individual gene trees (Figs S1–S4) were different to one another and to those in the combined tree, (apart from a few exceptions discussed below), the terminal clades were mostly consistent for the gene regions. To facilitate a discussion of the emerging results, the 24 lineages that represent genera or smaller groups were annotated using Roman numerals (I–XXIV). These were applied in the order of appearance in the concatenated tree (Fig. 5). Single species that grouped within major lineages, but not within defined species complexes, or not consistent within the same lineage in different trees, were labelled alphabetically (A–K).

Fig. 5.

Phylogenetic tree depicting the boundaries of currently accepted genera in the Ophiostomatales. This tree was generated using maximum likelihood analysis of the concatenated dataset of LSU, ITS, TEF1-α and RPBII gene regions. The dataset consisted of 264 isolates and 2 360 characters (including gaps). Bootstrap values above 60 % are shown. Bold lines indicate Bayesian posterior probabilities values above 0.8. Bootstrap values and Bayesian posterior probabilities values below the species complex level were removed for simplification. Purple blocks indicate existing genera, yellow blocks new genera described in this study, green blocks, genera that we redefine here, and blue blocks indicate genera that that have been reinstated and re-defined. (T = ex-type, E = ex-epitype, P = ex-paratype; L = ex-lectotype; A = authentic isolate, used in the original study; * Genome sequenced).

The overall topology of the LSU tree (Fig. S1) showed some differences from that of the concatenated tree, but 22 of the lineages corresponded between the two trees. The exceptions were Lineages III and XIX. Lineage III consisted of a single species in the concatenated tree, for which no LSU data were available, while Lineage XIX grouped outside the other genera in the concatenated tree, but as part of Sporothrix (Lineage XIV) in the LSU tree. Species complexes within Leptographium, Sporothrix and Ophiostoma were generally less well-defined in the LSU tree than in the concatenated tree.

The ITS tree (Fig. S2) showed little resolution below the genus level. Due to the variable nature of the ITS1 and ITS2 regions, the dataset was subjected to a strict Gblocks treatment (using automated parameters). The dataset consisted of 1 132 characters (including gaps) prior to treatment with Gblocks, and only 169 characters thereafter. The remaining dataset on which the tree (Fig. S2) is based, consisted predominantly of the 5.8S region. Nevertheless, we retained this analysis in the study because the ITS region is the officially recognised barcode for the fungi (Schoch et al. 2012). The ITS sequences were submitted to the GenBank Refseq database. The ITS tree (Fig. S2) supported separation of most of the genera, but Lineages XI, XII and XXII were not monophyletic in this tree, when compared to the concatenated tree. The relatively small dataset also failed to resolve most of the species complexes for the larger genera.

The TEF-1α tree (Fig. S3) resolved almost all the lineages representing species complexes, but failed to support monophyly of Lineages I, II, VIII, XI and XXII.

The RPBII tree (Fig. S4) supported the separation of all genera, species complexes and smaller lineages apart from Lineage XII that separated in four clades, and Lineage VIII that grouped within Lineage I.

Based on the concatenated tree (Fig. 5), Lineage I included species complexes previously defined in Leptographium s.l. (De Beer & Wingfield 2013), namely the Leptographium clavigerum, L. galeiforme, L. lundbergii, L. olivaceum, L. piceiperdum, L. procerum, L. serpens and L. wageneri complexes; as well as two species not forming part of these species complexes (A & B). Lineage II included the G. grandifoliae and G. penicillata species complexes and a smaller lineage (C). Leptographium piriforme was labelled as Lineage III and L. verrucosum as Lineage VI, both grouping outside of Leptographium (Lineage I). Lineage IV consisted of three isolates of the monotypic genus Esteya. The Raffaelea sulphurea complex formed Lineage V. Ceratocystiopsis spp. formed Lineage VII, Graphilbum formed Lineage VIII and Aureovirgo formed Lineage IX. Raffaelea vaginata grouped outside of Raffaelea (Lineage XII) and was labelled Lineage X. Lineage XI consisted of the R. lauricola complex, distinct from Raffaelea spp., which formed Lineage XII. The Ophiostoma tenellum complex formed Lineage XIII. Sporothrix and Ophiostoma, as defined by De Beer et al. (2016a), formed Lineages XIV and XV, respectively. Lineage XIV consisted of the S. gossypina and S. stenoceras species complexes (which grouped inseparably from each other), the S. candida, S. inflata and S. pallida species complexes, the pathogenic clade (including the type species of Sporothrix, S. schenckii), as well as groups D to G. Lineage XV included the O. clavatum, O. ips, O. minus, O. piceae, O. pluriannulatum and O. ulmi complexes as well as groups H to K. Sporothrix fumea and S. brunneoviolacea both grouped separate from Sporothrix, and were labelled as Lineages XVI and XIX, respectively. Three Ophiostoma species consistently grouped together and distinct from Ophiostoma and were labelled Lineage XVII. Lineage XVIII included O. valdivianum and Lineage XX consisted of O. denticulatum and O. angusticollis. Lineage XXI represented Fragosphaeria, and Lineage XXII represented Hawksworthiomyces. Ophiostoma noisomeae was labelled Lineage XXIII, while O. grandicarpum and O. microsporum together constituted Lineage XXIV.

Afroraffaelea was excluded from the final analyses because the placement of the type for this monotypic species, Afr. ambrosiae, was completely incongruent among the separate gene trees (data not shown), most often forming long branches, distinct from all other groups. This impacted negatively on the support for several lineages in the concatenated tree, which prompted the decision to exclude the species from the analyses. Likewise, the two species of Intubia and the monotypic Chrysosphaeria were excluded from the analyses due to their ambiguous generic placement when using traditionally applied phylogenetic markers (Nel et al. 2021).

TAXONOMY

Phylogenetic analyses for four gene regions revealed 16 lineages within the Ophiostomatales, which we now recognise as valid genera. Seven of these represent genera currently known and defined in the Order. They include Esteya (Lineage IV), Ceratocystiopsis (Lineage VII), Graphilbum (Lineage VIII), Aureovirgo (Lineage IX), Sporothrix (Lineage XIV), Fragosphaeria (Lineage XXI) and Hawksworthiomyces (Lineage XXII). Two Lineages (Lineage II and V), which were clearly distinct from all other genera, included the ex-type cultures of Grosmannia penicillata and Dryadomyces amasae respectively. These species have previously been treated in the genera Grosmannia/Leptographium and Raffaelea respectively and we have consequently reinstated and redefined the genera Grosmannia and Dryadomyces with emended descriptions. Four lineages (Lineage XI, XIII, XVII and XXIV) are recognised as representing new genera and are described as such. Characters of these new genera were previously included in the descriptions of Leptographium, Ophiostoma and Raffaelea, and we have consequently emended their descriptions. Although Afroraffaelea, Chrysosphaeria, Intubia, and the fossil genus Paleoambrosia were not included in our analyses, we recognise these genera as valid, retaining them in the Ophiostomatales.

The placement of the remaining lineages (III, VI, X, XVI, XVIII, XIX, XX and XXIII) remains uncertain, as most of these lineages were represented by single species that grouped inconsistently in our analyses. We have chosen not to describe new monotypic genera for these lineages, but rather to delay this decision until additional taxa are discovered that support establishing novel genera. In addition to describing and redefining genera, new combinations have been provided for species where necessary.

Circumscription of the Ophiostomatales and Ophiostomataceae

At present there is no need to revise the description of the Ophiostomatales. This is because the emended description by De Beer et al. (2013a) broadly encompasses the morphologies of all genera, including the novel genera described in the present study. With more than 300 species and clearly distinct broad morphological groups in the Order, it would make sense to provide a narrower definition for the Ophiostomataceae, and to introduce one or more additional families. However, in view of the lack of support for the deeper nodes in our analyses, we have refrained from doing so at present. We suggest that this should be done only when related taxa outside the Ophiostomatales, such as those included in the LSU and SSU phylogenies of De Beer et al. (2013a), can be incorporated in multigene phylogenies to provide more robust context within the Sordariomycetidae.

Currently accepted genera in the Ophiostomatales

All currently accepted genera in the Ophiostomatales are defined below, and based on our results, descriptions have been emended where necessary. Genera and species complexes are discussed in alphabetical order, with lineage numbers corresponding to their appearance in the concatenated phylogenetic tree (Fig. 5).

New combinations

Where required, new combinations have been provided, and these are listed under the relevant genera. Species that have been treated in a particular genus but were shown based on our data to reside in a different genus, for which a name already exists in the appropriate genus, have been listed under ‘current name’ in Table 1, and thus not in the following section For example, Grosmannia serpens is now treated as Leptographium serpens but did not require a new combination and is listed in Table 1.

Table 1.

Taxa described in the Ophiostomatales, based on currently published data.

Previous name	Current name	CMW1	CBS or other1	Type2	Isolated from	Country	Collector	GenBank Accession Numbers3
								ITS	LSU	TEF1-α	RPBII
Afroraffaelea
Afr. ambrosiae	Afr. ambrosiae	48331	141678	T	Premnobius cavipennis	Florida, USA	C. Bateman	OM632703	OM584293	OM631576	OM631577
Aureovirgo
A. volantis	A. volantis	41250	139649	P	Cyrtogenius africus on Euphorbia ingens	South Africa	J.A. van der Linde	OM501369	OM514700	OM631743	OM631579
A. volantis	A. volantis	38929	140081		Cyrtogenius africus on Euphorbia ingens	South Africa	J.A. van der Linde	OM501368	OM514699	OM631742	OM631578
Ceratocystiopsis
Cop. brevicomis	Cop. brevicomis	40952	333.97	T	Dendroctonus brevicomis	California, USA	T. Harrington	EU913722	EU913683	–	–
Cop. collifera	Cop. collifera	7074	126.89	T	Dendroctonus valens on Pinus teocote	Mexico	J. Marmolejo	EU913721	EU913681	–	–
Cop. concentrica	Cop. concentrica	–	WIN(M)71-07		Pinus banksiana	Canada	J. Reid, A. Olchowecki	–	AF135571	–	–
Cop. conicicollis	Cop. conicicollis	–	WIN(M)69-25		Abies balsemea	Canada	J. Reid, A. Olchowecki	–	–	–	–
Cop. longispora	Cop. longispora	–	UM48		Pinus sp.	Canada	A. Olchowecki	EU913723	EU913684	–	–
Cop. lunata #	Cop. lunata	55897	47171	T	Xylosandrus crassciusculus	South Africa	W.J. Nel	MW028169	MW028141	–	–
Cop. manitobensis	Cop. manitobensis	13792	UAMH9813	T	Manitoba beetle gallery in Pinus resinosa	Canada	J. Reid	EU913714	EU913674	–	–
Cop. minima	Cop. minima	162	182.86		Pinus banksiana	Wisconsin, USA	M.J. Wingfield	OM501370	OM514701	OM631744	–
Cop. minuta*	Cop. minuta	4352	138717		Ips cembrae	Poland	T. Kirisits	OM501372	OM514703	OM631745	OM631581
Cop. minuta-bicolor *	Cop. minuta-bicolor	1020	635.66	A	Gallery of Ips sp. in Pinus contorta	USA	R.W. Davidson	OM501371	OM514702	–	OM631580
Cop. neglecta	Cop. neglecta	22403	100596	I	Hylurgops palliatus	Germany	R. Kirschner	OM501373	OM514704	OM631746	OM631582
Cop. ochracea	Cop. ochracea	–	DAOM100148		Picea mariana	Canada	H.D. Griffin	–	–	–	–
Cop. pallidobrunnea	Cop. pallidobrunnea	–	UM51		Populus tremuloides	Canada	J. Reid	–	EU913682	–	–
Cop. parva	Cop. parva	43830	UAMH9650	T	Abies balsamea	Canada	A. Olchowecki	OM501374	–	OM631747	OM631583
Cop. ranaculosa	Cop. ranaculosa	13940	119683		Pinus echinata	North Carolina, USA	F. Hains	OM501375	OM514705	OM631748	OM631584
Cop. rollhanseniana	Cop. rollhanseniana	43831	UAMH9774	T	Unknown beetle on Pinus sylvestris	Norway	J. Reid	OM501376	OM514706	OM631749	–
Cop. spinulosa	Cop. spinulosa		DAOM110151	T	Tilia americana	Canada	H.D. Griffin	–	–	–	–
Cop. synnemata #	Cop. synnemata	–	NRIF 16918DA	T	Dryocoetes alni infesting Populus tremula	Poland	K. Miœkiewicz	MN900988	MN900988	MN901018	–
Cop. yantaiensis #	Cop. yantaiensis	–	SNM650	T	Pinus thunbergii	China	R. Chang	MW989411	MZ819924	MZ853080	–
Cop. weihaiensis #	Cop. weihaiensis	–	SNM649	T	Pinus thunbergii	China	R. Chang	MW989413	MZ819926	MZ853082	–
Chrysosphaeria
Chr. jan-nelii #	Chr. jan-nelii	47058	141570	T	Termitomyces fungal comb of Macrotermes natalensis	South Africa	W.J. Nel	MT637038	MT637006	–	–
Dryadomyces (R. sulphurea complex)
R. amasae	D. amasae	25542	116694	T	Amasa concitatus on Angiosperms	Taiwan	H. Gebhardt	–	MT629750	OM631750	OM631585
R. montetyi	D. montetyi	25537	463.94	T	Platypus cylindrus on Quercus suber	France	D. Vouland	–	MT629761	OM631751	–
R. quercivora *	D. quercivorus	36263	122982		Quercus mongolica	Japan	T. Kubono	MT633072	MT629762	OM631752	OM631586
R. quercus-mongolicae	D. quercus-mongolicae	37749	KACC44403	P	Quercus mongolica	South Korea	K.H. Kim	MT633073	MT629764	OM631754	OM631588
R. quercus-mongolicae *	D. quercus-mongolicae	37751	KACC44405		Platypus koryoensis-infested Quercus	South Korea	K.H. Kim	MT633074	MT629763	OM631753	OM631587
R. sulphurea*	D. sulphureus	25529	380.68		Xyleborus saxesenii gallery in Populus deltoides	Kansas, USA	L.R. Batra	MT633077	MT629768	OM631755	OM631589
Esteya
E. vermicola	E. vermicola	37427	100821		Olea europeae	Italy	S. Frisullo	–	OM514708	OM631757	OM631591
E. vermicola	E. vermicola	37428	156.82		Pinus sp.	Taiwan	T. Tatsuno	OM501378	OM514709	OM631758	–
E. vermicola*	E. vermicola	–	115803		S. intricatus and its galleries in oak trees	Czech Republic	L. Marvanova	OM501377	OM514707	OM631756	OM631590
Fragosphaeria
F. purpurea*	F. purpurea	48515	133.34	A	Fagus sp.	England, UK	C.G.C. Chesters	OM501379	OM514710	OM631759	OM631592
F. reniformis	F. reniformis	48516	134.34	A	Fagus sp.	England, UK	E.W. Mason	OM501381	–	OM631760	OM631593
Graphilbum
Gra. acuminatum #	Gra. acuminatum	54769	145828	T	Ips acuminatus gallery on Pinus sylvestris	Poland	R. Jankowiak	MN548902	–	MN548952	–
Gra. brunneocrinitum	Gra. brunneocrinitum	–	TRTC34581	T	Abies balsamea	Canada	–	–	–	–	–
Gra. carpaticum #	Gra. carpaticum	43141	145835	T	Pissodes piceae gallery on Abies alba	Poland	P. Majka	KY568116	–	MN548956	–
Gra. curvidentis #	Gra. curvidentis	54779	145832	T	Pitokteines curvidens gallery on Abies alba	Poland	P. Bilański	KY568111	–	KY56850	–
Gra. crescericum	Gra. crescericum	22828	130864	T	Hylurgops palliatus on Pinus radiata	Spain	P. Romón	OM501403	OM514749	OM631779	OM631604
Gra. curvicolle	Gra. curvicolle	–	WIN(M)70-25	T	Abies balsamea	Canada	J. Reid, A. Olchowecki	–	–	–	–
Gra. fragrans *	Gra. fragrans	19357	138720		Pinus patula	South Africa	X. D. Zhou	OM501404	OM514750	OM631780	OM631605
Gra. furuicola #	Gra. furuicola	44770	145813	T	Tomicus piniperda in Pinus sylvestris	Norway	R.H. Lindseth, T.H. Sundt	MN548907	–	MN548961	–
Gra. gorcense #	Gra. gorcense	34153	146203	T	Tetropium sp. in Picea abies	Poland	R. Jankowiak	MN548919	–	MN548972	–
Gra. interstitiale #	Gra. interstitiale	54780	145816	T	Hylurgops interstitialis in Pinus sylvestris	Russia	H. Solheim	MN548909	–	MN548963	–
Gra. ipis-grandicollis #	Gra. ipis-grandicollis	–	VPRI43762		Ips grandicollis gallery on Pinus radiata	Australia	A.J. Carnegie	MW046071	MW046117	MW066405	–
Gra. kesiyae	Gra. kesiyae	41729	139652	T	Polygraphus szemaoensis on Pinus kesiya	China	S. Taerum	MG205669	–	–	–
Gra. microcarpum	Gra. microcarpum	17196	YCC439	T	Cryphalus montanus	Japan	Y. Yamaoka	OM501405	OM514751	OM631781	OM631606
Gra. nigrum	Gra. nigrum	1097	163.61	A	Abies lasiocarpa	Colorado, USA	R.W. Davidson	OM501406	OM514752	–	OM631607
Gra. niveum #	Gra. niveum	–	SNM145	T	Pinus thunbergii	China	R. Chang	MW989418	–	MZ019548	–
Gra. puerense	Gra. puerense	41673	139640	T	Ips acuminatus on Pinus kesiya	China	S. Taerum	MG205671	–	–	–
Gra. rectangulosporium	Gra. rectangulosporium	29364	MAFF 238952	T	Polygraphus proximus on Abies mariesii	Japan	N. Ohtaka	MG205671	–	–	–
Gra. roseum #	Gra. roseum	40349	141074	T	Curtisia dentata	South Africa	T. Musvuugwa	KY050751	–	–	–
Gra. sexdentatum #	Gra. sexdentatum	54773	145814	T	Ips sexdentatus in Pinus sylvestris	Norway	H. Solheim, M.E. Waalberg	MN548915	–	MN548968	–
Gra. sparsum *	Gra. sparsum	37437	405.77	T	Bark beetle gallery on Picea glauca	Alaska, USA	R.W. Davidson	OM501409	OM514755	–	OM631608
Gra. translucens #	Gra. translucens	–	SNM144	T	Pinus thunbergii	China	R. Chang	MW989416	–	MZ019546	–
Gra. tsugae	Gra. tsugae	–	UAMH11701	T	Tsuga heterophylla	Canada	J. Reid, B. Reid	KJ661745	–	–	–
Gra. tubicolle	Gra. tubicolle	43837	UAMH9686	T	Pinus banksiana	Canada	A. Olchowecki	–	–	–	–
O. pusillum	Gra. pusilllum	14490	–	T	Pinus densiflora	Japan	H. Masuya	OM501407	OM514753	–	–
Grosmannia: G. penicillata complex
G. abiocarpa	G. abiocarpa	65	594.85	L	Ips sp. on Picea engelmannii	Colorado, USA	R.W. Davidson	OM501384	OM514715	OM631764	–
G. americana	G. americana	495	497.96	T	Dendroctonus simplex on Larix laricina	Vermont, USA	D. Bergdalh	OM501385	OM514718	OM631765	OM631595
G. aoshimae	G. aoshimae	29360	MAFF238948	T	Polygraphus proximus on Abies mariesii	Japan	N. Ohtaka	OM501386	OM514719	OM631766	OM631596
G. crassifolia #	G. crassifolia	38885	136505	T	Polygraphus poligraphus in Pinus crassifolia	China	X.D. Zhou, S. Taerum	–	MN644475	MN647897	–
G. dryocoetis	G. dryocoetis	473	376.66	T	Dryocoetes confusus on Abies lasiocarpa	Canada	A.C. Molnar	OM501392	OM514727	OM631768	OM631597
L. fenglinhense	G. fenglinhensis	44579	141896	T	Ips typographus on Pinus sp.	China	R. Chang, S.F. Chen	MH144128	–	MH124404	–
G. maixiuense	G. maixiuense	38884	136502	T	Polygraphus poligraphus, Ips shangrila in Picea crassifolia	China	M. Yin, S. Taerum, X.D. Zhou	OM501396	MN644474	MN647900	OM631599
G. penicillata *	G. penicillata	2644	116008		Picea abies	Norway	H. Solheim	OM501397	OM514737	OM631774	OM631600
G. purpurea	G. purpurea	38886	136975	T	Ips shangrila on Picea purpurea	China	M. Yin, S. Taerum, X.D. Zhou	OM501399	MN644476	MN647914	–
G. tibetensis	G. tibetensis	–	CFCC53415	T	Orthotomicus sp. on Pinus likiangensis var. balfouriana	Tibet	Z. Wang, Q. Lu	MT269759	–	MT268756	–
G. xeno-abietinum	G. xeno-abietinum	2410	136514	T	Pinus ponderosa	California, USA	T. Harrington	OM501402	MN644471	MN647894	OM631603
G. xianmiense #	G. xianmiense	38892	136500	T	Polygraphus poligraphus in Pinus crassifolia	China	X.D. Zhou, S. Taerum	–	MN644479	MN647911	–
G. zekuensis #	G. zekuensis	41876	141901	T	Bakerdania sp. in gallery of Ips nitidus on Picea crassifolia	China	S.J. Taerum	MH121683	MH121683	MH124546	–
L. abieticolens	G. abieticolens	2865	115248	T	Abies balsamea	Vermont, USA	D. Bergdalh	AF343701	–	–	–
L. abietinum	G. abietina	275	118590		Picea engelmannii	Canada	A. Molnar	OM501383	OM514713	OM631763	OM631594
L. altius	G. altior	12471	123619	T	Picea koraiensis	China	X.D. Zhou, Z.W. de Beer	HQ406851	–	HQ406875	–
L. bistatum	G. bistata	3804	120192	T	Pinus radiata	Japan	J.J. Kim	OM501388	OM514721	OM631795	–
L. chlamydatum	G. chlamydata	36631	128840		Pityogenes chalcographus on Picea abies	Finland	Z.W. de Beer	JF279965	–	JF280080	–
L. curviconidium	G. curviconidia	12441	123617	P	Ips typographus on Picea koraiensis	China	X.D. Zhou, Z.W. de Beer	OM501390	OM514725	OM631767	–
L. curvisporum	G. curvispora	17260	123914	T	Picea abies	Norway	M.J. Wingfield, H. Solheim	OM501391	OM514726	EU979347	–
L. engelmannii	= G. abietina	759	–		Picea engemannii	Canada	R.W. Davidson	–	OM631763	OM631762	–
L. eucalyptophilum	G. eucalyptophila	5211	–	A	Eucalyptus urophylla × E. pellita	Democratic Republic of Congo	J. Roux	OM501393	OM514728	OM631769	–
L. euphyes	G. euphyes	259	109701	T	Pinus strobus	New Zealand	M. Dick	AF343686	–	–	–
L. fruticetum	G. fruticetum	20605	–	T	Picea engelmannii × P. glauca	Canada	S. Massoumi Alamouti	OM501394	OM514730	OM631770	–
L. hughesii	G. hughesii	4052	109709	T	Aquilaria sp.	Vietnam	B. Lanchette	OM501395	OM514732	OM631772	OM631598
L. pistaciae	G. pistaciae	12499	123626	T	Pistacia chinensis	China	X.D. Zhou, Z.W. de Beer	HQ406846	–	HQ406870	–
Grosmannia: G. grandifoliae complex
G. grandifoliae	G. grandifoliae	703	ATCC28746	T	Fagus grandifolia	Iowa, USA	R.W. Davidson	–	–	OM631771	–
L. betulae	G. betulae	43191	142734	T	Scolytus ratzeburgi on Betula verrucosa	Poland	R. Jankowiak	KY801840	–	KY801817	–
L. pruni	G. pruni	10418	120197	P	Polygraphus ssiori on Prunus jamasakura	Japan	H. Masuya	OM501398	OM514740	OM631775	–
L. trypodendri	G. trypodendri	43182	142724	T	Trypodendron domesticum on Fagus sylvatica	Norway	R. Jankowiak	KY801828	–	KY801805	–
Grosmannia: Group C
G. abieticola	G. abieticola	17199	–	T	Dryocoetes hectographus on Abies mariesii	Japan	Y. Yamaoka	OM501382	OM514712	OM631761	–
L. innermongolicum	G. innermongolica	–	MUCL55158	T	Ips subelongatus on Larix sp.	China	Q. Lu	KM236107	–	KM981763	–
L. taigense	G. taigensis	36629	–	P	Ips typographus on Picea abies	Russia	Z.W. de Beer	OM501400	OM514744	OM631777	–
L. gestamen	G. gestamen	38096	CIEFAP453	T	Nothofagus dombeyi	Argentina	A. de Errasti	KT362234	KT362232	KT381300	–
Harringtonia gen. nov. (previously Raffalea lauricola complex)
R. aguacate	Har. aguacate	38067	141672	T	Persea americana	Florida, USA	C.L. Harmon	–	KJ909296	–	–
R. aguacate *	Har. aguacate	–	Raff. sp. 272		Persea americana	Florida, USA	C.L. Harmon	MT633065	MT629748	OM631783	OM631613
R. brunnea	Har. brunnea	–	378.68		Monarthrum sp.	USA	L.R. Batra	–	EU177457	–	–
R. lauricola *	Har. lauricola	–	Raff. sp. 570		Xyleborus sp. on Persea sp.	Florida, USA	J. Smith	MT633071	MT629759	OM631784	OM631614
R. lauricola	Har. lauricola	36261	PL159		Xyleborus glabratus	Georgia, USA	S. Fraedrich	OM501411	MT629760	OM631785	OM631615
Hawksworthiomyces
Haw. crousii	Haw. crousii	37531	MUCL55928	T	Bamboo chips	South Korea	J.J. Kim	KX396551	KX396548	OM652622	OM631609
Haw. hibbettii	Haw. hibbettii	37663	MUCL55929	T	Trachymyrmex sp.	Texas, USA	U. Mueller	KX396550	KX396547	OM652623	OM631610
Haw. lignivorus*	Haw. lignivorus	18600	119148	T	Eucalyptus pole	South Africa	E.M. de Meyer	OM501410	OM514756	OM631782	OM631611
Haw. taylorii	Haw. taylorii	20741	MUCL55927	T	Eucalyptus pole	South Africa	E.M. de Meyer	KX396549	KX396546	OM652624	OM631612
‘Haw. sequentia ENAS’	‘Haw. sequentia ENAS’	–	nik62104a_03C_19	T	Picea log	Sweden	–	HQ611296	–	–	–
Heinzbutinia gen. nov.
O. grandicarpum	He. grandicarpa	1600	250.88	T	Quercus robor	Poland	H. Butin	OM501412	OM514757	OM631786	OM631616
O. longicollum	He. longicolla	–	JCM10198	T	Quercus mongolica var. grosseserrata infested by Platypus quercivorus	Japan	H. Masuya	–	–	–	–
O. microsporum	He. microspora	17152	440.69	T	Quercus sp.	Virginia, USA	R.W. Davidson	–	OM514758	OM631787	OM631617
O. solheimii #	He. solheimii	52050	144881	T	Anisandrus dispar infesting Quercus robur	Poland	P.Wieczorek	MH283134	–	MH283488	–
Intubia
I. macrotermitinarum #	I. macrotermitinarum	46496	141560	T	Termitomyces fungal comb of Macrotermes natalensis	South Africa	W.J. Nel	MT637025	–	–	–
I. oerlemansii #	I. oerlemansii	47048	141564	T	Termitomyces fungal comb of Macrotermes natalensis	South Africa	W.J. Nel	MT637024	–	–	–
Jamesreidia gen. nov. (previously O. tenellum complex)
O. coronatum	J. coronata	43836	UAMH9685	T	Pinus sp.	Canada	A. Olchowecki	OM501413	OM514759	OM631788	OM631618
O. nigricarpum	J. nigricarpa	651	638.66		Pseudotsuga menziesii	Idaho, USA	R.W. Davidson	AY280490	DQ294356	–	–
O. rostrocoronata	J. rostrocoronata	456	434.77		Pulpwood chips	Wisconsin, USA	R.W. Davidson	AY194509	KX590871	–	–
O. tenellum	J. tenella	37439	189.86		Pinus sp.	Colorado, USA	R.W. Davidson	OM501414	OM514760	OM631789	–
Leptographium: L. clavigerum complex
G. aurea	L. aureum	667	438.69	A	Pinus contorta var. latifolia	Canada	R. W. Davidson	OM501387	OM514720	OM631793	OM631621
G. clavigera *	L. clavigerum	–	SL-Kw1407		Sapwood associated with Dendroctonus ponderosae	Canada	S. Lee	OM501421	OM514723	OM631799	OM631624
G. robusta	L. robustum	668	–	T	Pinus ponderosa	Idaho, USA	R.C.R. Jeffrey, R.W. Davidson	–	AY544619	JF798465	–
L. longiclavatum	L. longiclavatum	20608	–	P	Pinus contorta	Canada	S. Lee	–	OM514771	–	–
L. pyrinum	L. pyrinum	509	120181	T	Dendroctonus adjunctus	USA	R.W. Davidson	OM501445	OM514781	OM631819	OM631642
L. terebrantis	L. terebrantis	29841	337.70	T	Dendoctronus terebrantis	Louisiana, USA	S.J. Baras	JF798477	–	JF798470	–
L. tereforme	L. tereforme	37432	125736	T	Hylurgus ligniperda	California, USA	S.J. Kim	OM501455	OM514786	OM631828	OM631649
L. wingfieldii	L. wingfieldii	4741	–		Pinus densiflora	Japan	H. Masuya	OM501461	OM514789	OM631834	OM631654
Leptographium: L. galeiforme complex
G. galeiformis*	L. galeiforme	5290	115711	E	Pinus sylvestris	Scotland	T. Kirisits	OM501428	OM514731	OM631806	OM631631
G. radiaticola	L. radiaticola	9482	–		Hylurgus ligniperda on Pinus radiata	Chile	X.D. Zhou	OM501446	OM514742	OM631820	OM631643
L. doddsii #	L. doddsii	34479	143470	T	Dendroctonus valens	California, USA	M.J. Wingfield	MT637215	MT637212	MT637205	–
L. gordonii #	L. gordonii	34619	143477	T	Dendroctonus valens in Pinus resinosa	New Hampshire, USA	M.J. Wingfield	MT637226	MT637213	MT637207	–
L. koraiense	L. koraiense	44461	141898	T	Ips typographus on Pinus koraiensis	China	R. Chang	MH144096	–	MH124372	–
L. owenii #	L. owenii	34448	143467	T	Dendroctonus valens	California, USA	M.J. Wingfield	MT637217	KF515912.1	KF515884.1	–
L. seifertii #	L. seifertii	34620	143478	T	Dendroctonus valens on Pinus resinosa	New Hampshire, USA	M.J. Wingfield	MT637224	KF515911.1	KF515885.1	–
Leptographium: L. lundbergii complex
G. koreana	L. koreanum	14199	KUC2078	T	Tomicus piniperda on Pinus koraiensis	South Korea	J.J. Kim	OM501431	OM514733	OM631808	OM631633
G. yunnanensis	L. yunnanense	5152	–		Pinus yunnanensis	China	X.D. Zhou	AY707207	–	–	–
L. absconditum	L. absconditum	39763	136527	T	Orthotomicus laricis on Pinus nigra	Spain	P. Romón	OM501415	OM514761	OM631790	–
L. celere	L. celere	12422	123628	T	Pinus semaonensis	China	X.D. Zhou, Z.W. de Beer	HQ406834	–	HQ406858	–
L. conjunctum	L. conjunctum	12473	123631	T	Pinus yunnanensis	China	X.D. Zhou, Z.W. de Beer	HQ406831	–	HQ406855	–
L. lundbergii *	L. lundbergii	2190	138716		Pinus sylvestris	Norway	H. Roll-Hansen	OM501432	OM514772	OM631809	OM631634
L. manifestum	L. manifestum	12436	123622	T	Larix olgensis	China	X.D. Zhou, Z.W. de Beer	HQ406839	–	HQ406863	–
L. pinicola	L. pinicola	2398	–	T	Hylastes sp. on Pinus sp.	Canada	J. Juzwik	OM501439	OM514775	–	–
L. shansheni	L. shansheni	44462	141895	T	Ips typographus on Picea sp.	China	R. Chang, S.F. Chen	MH144097	–	MH124373	–
L. sosnaicola #	L. sosnaicola	52084	147023	T	Pinus sylvestris	Poland	D. Jazłowiecka	MT210337	MT210353	MT210397	–
L. truncatum	L. truncatum	28	929.85	T	Pinus taeda	South Africa	M.J. Wingfield	DQ062052/AY935626	–	DQ062019	–
L. wushanense	L. wushanense	–	YMF1.04936	T	Pinus sp.	China	J. Lu	MG878407	–	MG878409	–
Leptographium: L. olivaceum complex
G. cucullata *	L. cucullatum	1035	218.83	T	Ips typographus	Norway	H. Solheim	OM501423	OM514724	OM631801	OM631626
G. davidsonii	L. davidsonii	–	YCC611		Logs of Larix kaempferi infested with Ips subelongatus	Japan	Y. Yamaoka	GU134165	–	–	–
G. olivacea	L. olivaceum	23348	–		Pinus sylvestris	Finland	Z.W. de Beer, P. Niemelä	OM501434	OM514735	OM631811	–
G. olivaceapini	L. olivaceapini	116	504.86	E	Pinus ponderosa in Dendroctonus sp.	Arizona, USA	T. Hinds	OM501433	OM514736	OM631810	OM631635
G. sagmatospora	L. sagmatosporum	43846	UAMH6971		Pinus strobus	Canada	B. Grylls, K. Seifert	OM501449	OM514743	OM631823	–
G. vesca	L. vescum	34186	800.73		Ips pilifrons, Dendroctonus engelmanni in Picea engelmannii	Colorado, USA	F.F. Lombard, R. W. Davidson	OM501457	OM514745	–	–
L. brevicolle	L. brevicolle	–	150.78		Beetle gallery in Populus tremuloides	Colorado, USA	R.W. Davidson	MH055549	AF155670	MH055635	–
L. breviuscapum	L. breviuscapum	38888	136507	T	Picea crassifolia infested with Polygraphus poligraphus	China	M.L. Yin, X.D. Zhou	OM501419	OM514763	MN517742	–
L. conplurium	L. conplurium	23289	128834	P	Pinus sylvestris	Finland	Z.W. de Beer, P. Niemelä	OM501422	OM514765	OM631800	OM631625
L. duchongi	L. duchongi	44455	141897	T	Ips typographus on Pinus koraiensis	China	R. Chang	MH144122	–	MH124398	–
L. erubescens	L. erubescens	40672	278.54	E	Pinus sylvestris	Sweden	A. Mathiesen-Käärik	OM501425	OM514767	OM631803	OM631628
L. flavum	L. flavum	51797	144099	T	Quercus robur	Poland	R. Jankowiak	MH055548	–	MH055634	–
L. francke-grosmanniae	L. francke-grosmanniae	445	356.77	T	Hylecoetus dermestoides on Quercus sp.	Germany	H. Francke-Grosmann	OM501427	OM514768	OM631805	OM631630
L. sylvestris	L. sylvestris	23300	128833	P	Picea abies	Finland	Z.W. de Beer, P. Niemelä	OM501454	OM514777	OM631827	–
L. pseudoalbum	L. pseudoalbum	40671	276.54	T	Blastophagus piniperda in Pinus sylvestris	Sweden	A. Mathiesen-Käärik	MN516723	MN516723	MN517755	OM631641
L. raffai #	L. raffai	34451	143468	T	Dendroctonus valens	California, USA	M.J. Wingfield	MT637219	MT637211	MT637206	–
L. rhizoidum	L. rhizoidum	22809	136512	T	Hylastes ater on Pinus radiata	Spain	P. Romón, X.D. Zhou	MN516724	MN516724	MN517748	OM631644
L. tardum	L. tardum	51789	144091	T	Trypodendron domesticum on Fagus sylvatica	Poland	R. Jankowiak	MH055529	–	MH055615	–
L. vulnerum	L. vulnerum	51794	144096	T	Fagus sylvatica	Poland	R. Jankowiak	MH055534	–	MH055620	–
L. xiningense#	L. xiningense	38891	136509	T	Polygraphus poligraphus in Picea crassifolia	China	M.L. Yin, X.D. Zhou	MN516732	MN516732	MN517752	OM631637
Leptographium: L. piceiperdum complex
G. aenigmatica	L. aenigmaticum	2199	–	T	Ips typographus japonicus on Picea jezoensis	Japan	Y. Yamaoka	OM501416	OM514716	OM631791	OM631619
G. europhioides	L. europhioides	2811	115245		Picea rubens	New York, USA	T. Harrington	OM501426	OM514729	OM631804	OM631629
G. laricis	L. laricis	1913	120188		Larix sp.	Japan	Y. Yamaoka	–	DQ294393	–	–
G. piceiperdum *	L. piceiperdum	16768	138719		Picea glauca	Canada	K. Harrison	OM501435	OM514738	OM631812	OM631636
G. pseudoeurophioides	L. pseudoeurophioides	–	WIN(M)42		–	Canada	J. Reid	EU879136	–	–	–
L. heilongjiangense	L. heilongjiangense	44456	141702	T	Ips typographus on Pinus koraiensis	China	R. Chang	MH144098	–	MH124374	–
L. zhangii	L. zhangii	–	MUCL55162	T	Ips subelongatus on Larix gmelinii	China	X. Liu	KM236108	–	KM974275	–
Leptographium: L. procerum complex
L. bhutanense	L. bhutanense	18649	122076	T	Pinus wallichiana	Bhutan	M.J. Wingfield	OM501418	OM514762	OM631794	–
L. gracile	L. gracile	12398	123623	T	Pinus armandii	China	X.D. Zhou	OM501430	OM514769	–	–
L. latens	L. latens	12438	124023	T	Picea koraiensis	China	X.D. Zhou, Z.W. de Beer	HQ406845	–	HQ406869	–
L. longiconidiophorum	L. longiconidiophorum	2004	135624	T	Hylastes sp. on Pinus densiflora	Japan	M.J. Wingfield	KM491421	–	KM491471	–
L. peucophilum	L. peucophilum	2876	120191		Picea sp.	New York, USA	D. Bergdalh	–	–	–	–
L. pini-densiflorae	L. pini-densiflorae	5157	115261	T	Pinus densiflora	Japan	H. Masuya	OM501438	OM514774	OM631815
L. procerum *	L. procerum	34542	138288	E	Dendroctonus valens on Pinus resinosa	Maine, USA	M.J. Wingfield	OM501442	OM514778	OM631816	OM631639
L. profanum	L. profanum	10552	120307	T	Carya sp.	Alabama, USA	L. Eckhardt	OM501443	OM514779	OM631817	OM631640
L. sibiricum	L. sibiricum	4481	115260	T	Monochamus urussoni on Abies sibitica	Russia	V.P. Vetrova	OM501451	OM514783	–	–
L. sinense	L. sinense	38171	316515	P	Pinus elliottii	China	M. Yin, R. Chang, X.D. Zhou	OM501452	OM514784	OM631825	OM631647
L. sinoprocerum	L. sinoprocerum	29990	MUCL46532	T	Pinus tabuliformis	China	Q. Lu	OM501453	OM514785	OM631826	OM631648
L. yichunense	L. yichunense	44464	141705	T	Ips typographus on Picea sp.	China	R. Chang, S.F. Chen	MH144114	–	MH124390	–
Leptographium: L. serpens complex
G. alacris	L. alacre	621	128830	T	Pinus pinaster	Portugal	M. de Famtima Moniz	OM501417	OM514717	OM631792	OM631620
G. serpens	L. serpens	305	141.36	T	Pinus sylvestris	Italy	G. Goidànich	OM501450	–	OM631824	OM631646
L. castellanum	L. castellanum	2320	128698	P	Pinus occidentalis	Dominican Republic	R. Webb	OM501420	OM514764	OM631798	OM631623
L. gibbsii	L. gibbsii	36371	–	T	Hylastes opacus on Pinus sylvestris	England, UK	J. Gibbs	OM501429	–	OM631807	OM631632
L. rhodanense	L. rhodanense	16438	138284	T	Pinus sylvestris	Switzerland	U. Heiniger	OM501448	–	OM631822	OM631645
L. yamaokae	L. yamaokae	4726	129732	T	Pinus densiflora	Japan	H. Masuya	–	–	OM631835	OM631655
Leptographium: L. wageneri complex
G. wageneri *	L. wageneri var. ponderosum	279	–		Pinus sp.	USA	T. Harrington	OM501458	OM514746	OM631831	OM631651
L. douglasii *	L. douglasii	2076	–	P	Pseudotsuga menziesii	New Mexico, USA	M. Midke	OM501424	OM514766	OM631802	OM631627
L. neomexicanum	L. neomexicanum	2079	168.93	T	Pinus ponderosa	New Mexico, USA	T. Harrington, W Livingston	AY553382	–	AY536176	–
L. reconditum	L. reconditum	15	116348		Zea mays	South Africa	W. Jooste	AF343690	–	AY536177	–
L. wageneri var. pseudotsugae*	L. wageneri var. pseudotsugae	154	115246		Pseudotsuga menziesii	USA	T. Harrington	OM501459	OM514747	OM631832	OM631652
L. wageneri var. wageneri*	L. wageneri var. wageneri	53	139665		Pinus ponderosa	California, USA	T. Harrington	OM501460	OM514788	OM631833	OM631653
Leptographium: Group A
L. pineti	L. pineti	3837	115257		Pinus sp.	Indonesia	M.J. Wingfield	–	–	OM631814	OM631638
L. ningerense	L. ningerense	41786	139663	T	Coccotrypes cyperi on Pinus kesiya	China	S. Taerum	MG205674	–	MG205765	–
Leptographium: Group B
G. cainii	L. cainii	24907	–		Picea sp.	Canada	C. Breuil	OM501389	OM514722	OM631797	OM631622
Leptographium Incertae sedis (based on our data)
G. huntii	G. huntii	2868	118780		Pinus strobus	North Carolina, USA	V. Lackner	AY553394	–	DQ354938	–
G. leptographioides	G. leptographioides	481	144.59		Quercus sp.	New York, USA	R.W. Davidson	–	DQ294382	–	–
G. truncicola	G. truncicola	–	–		Dendroctonus sp. on Picea sp.	USA	–	–	–	–	–
L. albopini	L. albopini	^26	–		Hylastes sp. on Pinus sp.	USA	–	AF343695	–	–	–
L. alethinum	L. alethinum	^3763	–		Galleries of Hylobus abietis on log of Pinus nigra var. maritima	England, UK	A. Uzunovic	AY553391	–	AY536185
L. calophylli	L. calophylli	752	277.51		Cryphalus sp. on Calophyllum sp.	Mauritius	J.A. Stevenson	MH856855	MH868375	–	–
L. microsporum	L. microsporum	–	–	T	Fagus sp.	Mississippi, USA	R.W. Davidson	–	–	–	–
L. obscurum	L. obscurum	37429	125.39		Pinus sp.	USA	R.W. Davidson	–	–	–	–
L. pityophilum	L. pityophilum	2840	109706		Pinus nigra	Italy	S. Frisullo	OM501441	OM514776	–	–
L. rostrocylindricum	L. rostrocylindricum	–	–		Quercus sp.	Connecticut, USA	R.W. Davidson	–	–	–	–
Leptographium & Grosmannia incertae sedis (based on our data)
L. guttulatum	L. guttulatum	1310	120185	T	Tomicus piniperda on Pinus sylvestris	England, UK	J. Gibbs	–	OM514770	–	–
Lineage III
G. crassivaginata	G. crassivaginata	90	120178		–	–	T. Hinds	AF343673	–	–	–
L. alneum#	L. alneum	52076	144901	T	Dryocoetes alni infesting Populus tremula	Poland	K. Miœkiewicz	MN900997	MN900997	MN901024	–
L. piriforme	L. piriforme	25381	UAMH10681	P	Beetle caught in a trap baited with coyote dung	Canada	M.D. Greif	OM501440	–	–	–
Lineage VI
L. verrucosum	L. verrucosum	17160	112420	T	Xyleborus dryographus	Germany	H. Gebhardt	OM501456	OM514787	OM631830	OM631650
Masuyamyces gen. nov.
O. ambrosium	M. ambrosius	1024	210.64		Wood of Pinus sylvestris	Netherlands	J.A. von Arx	OM501465	OM514793	OM631836	OM631656
O. acarorum	M. acarorum	41850	139748	T	Orthotomicus angulatus on Pinus kesiya	China	S. Taerum	MG205657	–	–	–
O. botuliforme	M. botuliformis	14493	–		Cryphalus jeholensis	Japan	H. Masuya	OM501471	OM514799	OM631837	–
O. jilinense	M. jilinensis	40491	141894	T	Ips typographus on Picea sp.	China	X. D. Zhou	MH144094	–	MH124370	–
O. lotiforme #	M. lotiformis	–	MUCL55165	T	Ips subelongatus on Pinus sylvestris var. mongolica	China	X. Meng	MK748185	–	–	–
O. massoniana	M. massonianae	–	MUCL55179	T	Monochamus sp. on Pinus sp.	China	Q. Lu	KY094067	–	–	–
O. pallidulum	M. pallidulus	23278	128118	T	Pinus sylvestris	Russia	Z.W. de Beer, P. Niemelä	HM031510	–	–	–
O. saponiodorum	M. saponiodorus	28135	128302		Pinus sylvestris	Russia	R. Linnakoski	OM501512	OM514838	OM631838	OM631657
Ophiostoma: O. clavatum complex
O. ainoae	O. ainoae	1037	205.83	T	Picea abies	Norway	H. Solheim	OM501463	OM514791	–	–
O. brevipilosi	O. brevipilosi	41873	139660	T	Tomicus brevipilosus on Pinus kesiya	China	S. Taerum	MG205660	–	MG205732	–
O. brunneociliatum	O. brunneociliatum	5212	–		Larix sp.	Scotland, UK	T. Kirisits	KU184422	–	KU184379	–
O. brunneolum	O. brunneolum	23145	–	A	Picea abies	Russia	J. Ahtiainen, P. Niemelä	OM501472	OM514800	OM631843	OM631663
O. clavatum	O. clavatum	37983	141080	E	Ips acuminatus on Pinus sylvestris	Sweden	C. Villari	OM501477	OM514805	OM631848	–
O. hongxingense #	O. hongxingense	–	CFCC52695	T	Ips subelongatus on Larix gmelinii	China	Q. Lu	MK748194	–	MN896068	–
O. japonicum	O. japonicum	2202	YCC099	T	Ips typographus japonicus on Picea jezoensis	Japan	Y. Yamaoka	OM501492	–	OM631855	OM631673
O. jiamusiensis	O. jiamusiensis	40512	141893	T	Ips typographus on Picea sp.	China	X.D. Zhou	MH144064	–	MH124343	–
O. macroclavatum	O. macroclavatum	23115	141081	T	Pinus sylvestris	Russia	Z.W. de Beer	HM031499	–	KU094765	–
O. peniculi #	O. peniculi	–	CFCC52687	T	Ips subelongatus infesting Larix gmelinii	China	Q. Lu	MK748198	–	MN896063	–
O. poligraphi	O. poligraphi	38899	136517	T	Polygraphus poligraphus on Picea crassifolia	China	M. Yin, S. Taerum, X.D. Zhou	OM501507	OM514832	OM631871	OM631688
O. pseudocatenulatum	O. pseudocatenulatum	43103	141276	T	Ips cembrae on Larix decidua	Poland	R. Jankowiak	KU094686	–	KU094774	–
O. shangrilae	O. shangrilae	38901	136519	T	Ips shangrila on Picea purpurea	China	M. Yin, S. Taerum, X.D. Zhou	OM501514	OM514840	OM631879	OM631695
O. songshui	O. songshui	44473	141707	T	Ips typographus on Picea sp.	China	R. Chang, S.F. Chen	MH144065	–	MH124344	–
O. subelongati #	O. subelongati	–	CFCC52693	T	Ips subelongatus infesting Larix gmelinii	China	Q. Lu	MK748200	–	MN896064	–
O. tapionis	O. tapionis	23266	128122	P	Picea abies	Russia	Z.W. de Beer, P. Niemelä	OM501516	OM514842	OM631881	OM631697
Ophiostoma sp. 3 (Hyalorhinocladiella sp. 2)	Ophiostoma sp. 3 (Hyalorhinocladiella sp. 2)	43851	UAMH10642		Ips sp.	Canada	S. Massoumi Alamouti	OM501524	OM514851	OM631888	–
Ophiostoma sp. 4 (Hyalorhinocladiella sp. 1)	Ophiostoma sp. 4 (Hyalorhinocladiella sp. 1)	43848	UAMH10639		Ips sp.	Canada	S. Massoumi Alamouti	OM501525	OM514852	OM631889	OM631704
Ophiostoma: O. ips complex
O. adjuncti	O. adjuncti	1025	314.77	T	Stained sapwood in Pinus ponderosa	New Mexico, USA	R.W. Davidson	OM501462	OM514790	–	OM631658
‘O. arborea’	‘O. arborea’	–	WIN(M)69-23		Picea mariana	Canada	A. Olchowecki, J. Reid	–	–	–	–
O. bicolor	O. bicolor	23169	–		Pinus sylverstris	Russia	J. Ahtiainen, P. Niemelä	OM501470	OM514798	OM631842	OM631662
O. columnare	O. columnare	–	WIN(M)71-27		Pinus banksiana	Canada	A. Olchowecki, J. Reid	–	–	–	–
O. fuscum	O. fuscum	23195	128124	P	Pinus sylverstris	Finland	Z.W. de Beer, P. Niemelä	OM501483	OM514810	–	OM631670
O. gilletteae #	O. gilletteae	30681	143458	T	Dendroctonus valens	Washington, USA	N. Gillette	MT637227	–	–	–
O. guatemalensis	O. guatemalensis	44221	–	T	Pinus patula	Guatemala	I. Barnes, J. Garnas	–	–	–	–
O. hyalothecium	O. hyalothecium	–	ATCC28825		Pinus contorta	Wyoming, USA	R.W. Davidson	–	AF137284	–	–
Tuberculariella ips	O. ips-like sp. 6	14175	435.34	T	Ips sp. on Pinus sp.	Minnesota, USA	J.G. Leach	OM501487	OM514813	OM631854	OM631672
O. ips *	O. ips	19371	138721		Pinus taeda	Louisiana, USA	X.D. Zhou	OM501486	OM514812	OM631853	OM631671
O. manchongi #	O. manchongi	41954	141906		Uropodoidea sp. in Ips shangrila gallery on Picea purpurea	China	S.J. Taerum	MH121662	–	–	–
O. montium	O. montium	15419	–		Pinus contorta	Idaho, USA	B. Bentz	OM501498	OM514822	OM631861	OM631678
O. pseudobicolor #	O. pseudobicolor	–	CFCC52683	T	Ips subelongatus in Larix gmelinii	China	Q. Lu	MK748188	–	–	–
O. pulvinisporum	O. pulvinisporum	9022	118673	T	Pinus pseudostrobus	Mexico	X.D. Zhou	OM501509	OM514835	OM631874	OM631691
Ophiostoma: O. minus complex
‘O. album’	‘O. album’	–	MUCL55189	T	Monochamus alternatus gallery on Pinus massoniana	China	Q. Lu, Y.Y. Lun	KY094073	–	–	–
O. exiguum	O. exiguum	–	–		Pinus virginiana	West Virginia, USA	G.G. Hedgcock	–	–	–	–
O. kryptum	O. kryptum	–	116190	T	Tetropium on Picea	Austria	T. Kirisits	AY305685	–	–	–
O. minus	O. minus	43873	UAMH4917		Dendroctonus ponderosae on Pinus flexillis	Canada	P. Muruyama	OM501497	OM514821	OM631860	OM631677
‘O. olgensis’	‘O. olgensis’	–	CXY1410	T	Ips subelongatus on Larix olgensis	China	Q. Lu	KU551303	–	KU551297	–
O. minus (in Europe O. pini)	O. minus (in Europe O. pini)	43346	–		Pinus sylvestris	Poland	T. Tomasz	–	–	–	–
O. pseudominus	O. pseudominus	43878	UAMH9721		Pseudotsuga menziesii	Canada	J. Reid, B Reid	OM501508	OM514834	OM631873	OM631690
O. pseudotsugae	O. pseudotsugae	–	D48/3		–	Canada	H. Solheim	AY542501	–	–	–
O. wuyingense	O. wuyingense	44474	141706	T	Ips typographus on Picea sp.	China	R. Chang, S.F. Chen	MH144061	–	MH124340	–
Ophiostoma: O. piceae complex
O. arduennense (= O. distortum)	O. distortum	40266	MUCL44866	T	Fagus sylvatica	Belgium	F.X. Carlier, T. Defrance	OM501468	OM514796	KU184376	–
O. brunneum	O. brunneum	1027	161.61	A	Standing dead Abies sp.	Colorado, USA	R.W. Davidson	OM501473	OM514801	OM631844	OM631664
O. canum (Pachnodium)	O. canum (Pachnodium)	5023	118668		Tomicus minor on Pinus sylvestris	Austria	T. Kirisits	OM501474	OM514802	OM631845	OM631665
O. cupulatum	O. cupulatum	37441	102358	T	Pseudotsuga menziesii	Washington, USA	T. Harrington	OM501479	OM514806	OM631849	OM631667
O. distortum	O. distortum	467	397.77		Picea engelmannii	Arizona, USA	R.W. Davidson	OM501481	OM514808	OM631850	OM631668
O. flexuosum	O. flexuosum	907	208.83	T	Picea abies	Norway	H. Solheim	OM501482	OM514809	OM631851	OM631669
O. floccosum	O. floccosum	34182	799.73	T	Wood	Sweden	A. Mathiesen-Käärik	KU184431	–	KU184388	–
O. genhense #	O. genhense	–	CFCC52675	T	Ips subelongatus infesting Larix gmelinii	China	Q. Lu	MK748199	–	MN896074	–
O. kunlunense #	O. kunlunense	41927	141903	T	Uropodoidea sp. in gallery of Ips shangrila on Picea purpurea	China	S.J. Taerum	MH121648	–	MH124515	–
O. micantis	O. micantis	38903	136523	T	Dendroctonus micans on Picea crassifolia	China	M. Yin, S. Taerum, X.D. Zhou	OM501496	OM514820	OM631859	OM631676
O. multisynnematum #	O. multisynnematum	–	CFCC52677		Ips subelongatus infesting Larix gmelinii	China	Q. Lu	MK748196	–	MN896071	–
O. nikkoense	O. nikkoense	17193	YCC430	T	Polygraphus proximus	Japan	Y. Yamaoka	OM501500	OM514824	OM631863	OM631679
O. nitidi	O. nitidi	38907	136525	T	Picea crassifolia	China	M. Yin, S. Taerum, X.D. Zhou	OM501501	OM514825	OM631864	OM631680
O. perfectum	O. perfectum	17153	600.85		Cat hairs	Germany	G.S. de Hoog	OM501506	OM514831	OM631868	OM631685
O. peregrinum	O. peregrinum	–	CIEFAP426	T	Pinus radiata	Argentina	A. de Errasti	MG345116	–	–	–
O. piceae *	O. piceae	–	UAMH11346		Pine saw timber	Canada	A. Uzunovic	MT633062	MT629745	OM631869	OM631686
O. pityokteinis #	O. pityokteinis	52056	144879	T	Pityokteines curvidens infesting Abies alba	Poland	P. Majka	MH837046	–	MH837055	–
O. qinghaiense	O. qinghaiense	38902	136521	T	Picea crassifolia	China	M. Yin, S. Taerum, X.D. Zhou	OM501510	OM514836	OM631875	–
O. rachisporum	O. rachisporum	28021	–		Picea abies	Russia	R. Linnakoski	OM501511	OM514837	OM631877	OM631693
O. rufum #	O. rufum	52062	144871	T	Ips cembrae galleries on Larix decidua	Czech Republic	K. Lukášová	MH837040	–	KY568647	–
O. setosum	O. setosum	27833	AU160-38	T	Tsuga sp.	Canada	A. Uzunovic	OM501513	OM514839	OM631878	OM631694
O. shanziensis #	O. shanziensis	48329	MUCL46456	T	Phloem adjacent to Dendroctonus valens gallery in Pinus tabuliformis	China	Q. Lu, C. Decock	MT637221	–	–	–
O. sugadairense	O. sugadairense	–	YCC589	T	Polygraphus sp. on Larix sp.	Japan	Y. Yamaoka	LC090227	–	AB934343	–
O. taphrorychi #	O. taphrorychi	52045	144891	T	Taphrorychus bicolor infesting Fagus sylvatica	Poland	P. Bilánski	MH837052	–	MH837062	–
O. typographi	O. typographi	44483	141709	T	Picea sp.	China	R. Chang, S.F. Chen	MH144059	–	MH124337	–
O. torulosum	O. torulosum	10574	CTK106		Fagus sylvatica	Germany	T. Kirisits	OM501518	OM514844	OM631883	OM631699
O. xinganense #	O. xinganense	–	CFCC52679		Ips subelongatus infesting Larix gmelinii	China	Q. Lu	MK748186	–	MN896078	–
Ophiostoma: O. pluriannulatum complex
O. californicum	O. californicum	143	796.73		Prunus domestica	California, USA	R.W. Davidson	KU756602	AF137280	–	–
O. carpenteri	O. carpenteri	44611	UAMH9696	P	Trypodendron lineatum	Oregon, USA	S. Carpenter	OM501475	OM514803	OM631846	–
O. conicola	O. conicola	2561	127.89	T	Pinus cembroides	Mexico	H. Butin	OM501478	–	–	–
O. longiconidiatum	O. longiconidiatum	14265	121350	A	Faurea saligna	South Africa	G. Kamgan Nkuekam	OM501494	OM514818	OM631857	–
O. longirostellatum	O. longirostellatum	–	134.51	T	Quercus sp.	Scotland, UK	B.K. Bakshi	–	AF155688	–	–
O. multiannulatum	O. multiannulatum	2567	357.77		Pinus sp.	North Carolina, USA	R.W. Davidson	OM501499	OM514823	OM631862	–
O. novae-zelandiae	O. novae-zelandiae	–	CIEFAP423	T	Dead fallen wood	New Zealand	–	KT362249	KT362226	–	–
O. palustre	O. palustre	44403	140596	T	Barringtonia racemosa	South Africa	J.A. Osorio	KU865593	–	–	–
O. pluriannulatum	O. pluriannulatum	–	MUCL18372		Quercus sp.	Oregon, USA	R.W. Davidson	AY934517	DQ294365	–	–
O. populinum	O. populinum	50149	212.67	T	Populus tremuloides	Colorado, USA	R.W. Davidson	–	OM514833	OM631872	OM631689
O. retusi	O. retusi		ATCC22324		Populus sp.	Colorado, USA	R.W. Davidson	–	L05783	–	–
O. sparsiannulatum	O. sparsiannulatum	17231	122815	T	Pinus taeda	Georgia, USA	L. Eckhardt	OM501515	OM514841	OM631880	OM631696
O. subannulatum	O. subannulatum	518	188.86		Pinus sp.	USA	W.H. Livingston, R.W. Davidson	AY934522	DQ294364	–	–
Ophiostoma: O. ulmi complex
O. allantosporum	O. allantosporum	163	185.86		Pinus resinosa	Wisconsin, USA	M.J. Wingfield	OM501464	OM514792	OM631839	OM631659
O. araucariae	O. araucariae	40665	114.68	T	Araucaria araucana	Chile	H. Butin	OM501467	OM514795	–	–
O. australiae	O. australiae	6606	121025	T	Acacia mearnsii	Australia	M.J. Wingfield	EF408603	–	–	–
O. bacillisporum	O. bacillisporum	2579	MUCL45378	T	Xyloterus domesticus on Fagus sylvatica	Germany	F.X. Carlier	OM501469	OM514797	OM631841	OM631661
O. borealis	O. borealis	18966	123222	T	Betula logs	Norway	G. Kamgan Nkuekam, H. Solheim	EF408593	–	–	–
O. catonianum	O. catonianum	11535	263.35	A	Pyrus communis	Italy	G. Goidanich	AF198243	–	–	–
O. denticiliatum	O. denticiliatum	29493	124497	T	Betula pendula	Norway	R. Linnakoski	FJ804490	–	–	–
O. himal-ulmi	O. himal-ulmi	22729	ATCC36204		Ulmus sp.	India	H. Rebel	OM501484	OM514811	OM631852	–
O. hylesini	O. hylesini	51680	144296	T	Hylesinus crenatus on Fraxinus excelsior	Poland	P. Wieczorek	MH055636	MH055675	MH062835	–
O. introcitrinum	O. introcitrinum	–	UAMH9549	T	Betula sp.	Canada	A. Olchowecki	OM501485	–	–	–
O. karelicum	O. karelicum	23099	123219	T	Scolytus sp. on Betula sp.	Russia	Z.W. de Beer, P. Niemelä	OM501493	OM514817	OM631856	OM631674
O. novo-ulmi subsp. americana	O. novo-ulmi subsp. americana	43874	UAMH5030		Ulmus americana	–	–	–	OM514828	–	OM631683
O. novo-ulmi subsp. novo-ulmi*	O. novo-ulmi subsp. novo-ulmi	–	H327		Ulmus sp.	Slovakia	H. Jamnicky	OM501503	OM514827	OM631865	OM631682
O. novo-ulmi subsp. novo-ulmi	O. novo-ulmi subsp. novo-ulmi	43870	UAMH10443		Ulmus sp.	Iran	M. Rahju, K. Rahnama	OM501504	OM514829	OM631866	OM631684
O. patagonicum	O. patagonicum	38089	CIEFAP431	T	Nothofagus pumilio	Argentina	A. de Errasti	OM501505	OM514830	OM631867	–
O. pseudokarelicum	O. pseudokarelicum	51704	144281	T	Trypodendron domesticum on Alnus incana	Norway	T. Aas	MH055659	MH055693	MH062859	–
O. quercus *	O. quercus	2465	117912		Quercus robur	France	M. Morelet	MT633064	MT629747	OM631876	OM631692
O. signatum	O. signatum	51689	144269	T	Trypodendron signatum on Alnus incana	Norway	G. Kvammen	MH055645	MH055682	MH062844	–
O. tasmaniense	O. tasmaniense	29088	127212	T	Eucalyptus nitens stumps	Australia	G. Kamgan Nkuekam	GU797211	–	GU797223	–
O. tsotsi	O. tsotsi	18134	123599	A	Julbenardia globiflora	Malawi	J. Roux	OM501520	OM514846	–	OM631701
O. ulmi *	O. ulmi	–	W9		Ulmus sp.	–	–	OM501521	OM514847	OM631885	OM631702
O. undulatum	O. undulatum	19396	127183		Eucalyptus grandis	Australia	M.J. Wingfield	–	OM514848	–	–
O. villosum	O. villosum	51694	144274	T	Dryocoetes villosus on Quercus robur	Norway	T. Aas, K.D. Hansen	MH055650	MH055685	MH062849	–
Ophiostoma: Group H
O. triangulosporum	O. triangulosporum	1033	138.77	T	Araucaria angustifolia	Brazil	H. Butin	OM501519	OM514845	OM631884	OM631700
Ophiostoma: Group I
O. macrosporum *	O. macrosporum	14176	367.53	T	Pinus sylvestris	Sweden	H. Francke-Grosmann	OM501495	OM514819	OM631858	OM631675
O. tingens	O. tingens	25530	366.53		Xyleborus saxesenii gallery in Populus deltoides	Sweden	H. Francke-Grosmann	–	EU177474	–	–
Ophiostoma sp. 1 (Ambrosiella sp. 1)	Ophiostoma sp. 1 (Ambrosiella sp. 1)	43827	UAMH10633		Ips sp.	Canada	S. Massoumi Alamouti	OM501522	OM514850	OM631887	OM631703
Ophiostoma sp. 2 (Ambrosiella sp. 2)	Ophiostoma sp. 2 (Ambrosiella sp. 2)	43828	UAMH10634		Ips sp.	Canada	S. Massoumi Alamouti	OM501523	–	–	–
Ophiostoma: Group J
O. piliferum *	O. piliferum	–	–		–	–	–	MT633063	MT629746	OM631870	OM631687
O. ponderosae	O. ponderosae	37953	ATCC26665	T	Blue stain of Pinus ponderosa	Arizona, USA	R.W. Davidson	OM501554	OM514882	–	–
Ophiostoma: Group K
O. tetropii	O. tetropii	4470	428.94		Picea abies	Austria	T. Kirisits	OM501517	OM514843	OM631882	OM631698
Ophiostoma incertae sedis (based on our data)
Lineage XVIII
O. valdivianum	O. valdivianum	449	454.83	T	Nothofagus alpiva	Chile	H. Butin	–	OM514849	OM631886	–
Lineage XX
O. angusticollis	O. angusticollis	152	–		Pinus banksiana	Wisconsin, USA	M.J. Wingfield	OM501466	OM514794	OM631840	OM631660
O. denticulatum	O. denticulatum	146	ATCC38087	T	Gnathotrichus sp. on Pinus sp.	Colorado, USA	R.W. Davidson	OM501480	OM514807	–	–
O. sejunctum	O. sejunctum	–	Ophi 1A	T	Tomicus sp. on Pinus sp.	Spain	M.R. Bareal	AY934519	–	–	–
Lineage XXIII
O. noisomeae	O. noisomeae	40326	–	T	Rapanea melanophloeos	South Africa	T. Musvuugwa	OM501502	OM514826	–	OM631681
Paleoambrosia
P. entomophila	P. entomophila	–	No. B-F-7	T	Palaeotylus femoralis in burmese amber	Myanmar	G.O. Poinar, F.E. Vega	–	–	–	–
Raffaelea
R. albimanens*	R. albimanens	25532	271.70	T	Platypus externedentatus in Ficus sycomorus	South Africa	D.B. Scott	MT633066	MT629749	OM631890	OM631705
R. ambrosiae*	R. ambrosiae	25533	185.64	T	Platypus cylindrus tunnel in Quercus sp.	England, UK	J.M. Bakeer	MT633067	MT629751	OM631891	OM631706
R. arxii	R. arxii	25534	273.70		Xyleborus torquatus on Cussonia sp.	South Africa	D.B. Scott	–	MT629754	OM631892	OM631708
R. borbonica #	R. borbonica	51553	PPRI27953	T	Leucaena leucocephala	Réunion, France	M.J. Wingfield, P.W. Crous	MT633059	MT629742	–	–
R. campbelliorum	R. campbelliorum	44800	139943	T	Xyleborus glabratus on Persea palustris	Florida, USA	A.S. Campbell	–	KR018414	–	–
R. canadensis	R. canadensis	25536	168.66	T	Platypus wilsonii in Pseudotsuga menziessii	Canada	A. Funk	GQ225699	MT629755	–	–
R. canadensis (=A. sulcati)	R. canadensis	25528	805.70	T	Gmnathotrichus sulcatus (mycangium) on Pseudotsuga menziesii	Canada	A. Funk	–	EU177459	–	–
R. crossotarsi	R. crossotarsi	44793	141675	T	Mycangium extract of Crossotarsus emancipatus in Lithocarpus sp.	Taiwan	J. Hulcr, A. Black, D.R. Simmons	KX267138	MT629756	OM631893	OM631709
R. cyclorhipidii	R. cyclorhipidii	44790	141676	T	Cyclorhipidion sp. on Lithocarpus sp.	Taiwan	J. Hulcr, A. Black, D.R. Simmons	MT633069	MT629757	–	OM631710
R. ellipticospora	R. ellipticospora	38056	121569	T	Xyleborus glabratus on Persea sp.	South Carolina, USA	S. Fraedrich	MT633070	MT629758	OM631894	–
R. fusca	R. fusca	38798	121570	T	Xyleborus glabratus on Persea sp.	South Carolina, USA	S. Fraedrich	–	EU177449	–	–
R. gnathotrichi	R. gnathotrichi	25523	379.68	T	Gnathotrichus retusus on Picea engelmannii	Colorado, USA	L.R. Batra	–	EU177460	–	–
R. promiscua #	R. promiscua	55899	147173	T	Xyleborinus saxesenii	South Afica	W.J. Nel	MW028176	–	–	–
R. rapaneae	R. rapaneae	40357	140084	T	Platypodinae sp. on Rapanea melanophloeos	South Africa	T. Musvuugwa	KT192596	KT182930	–	–
R. santoroi	R. santoroi	25539	399.67	T	Platypus sp. bore hole	Argentina	J. Wright	MT633075	MT629765	–	–
R. scolytodis	R. scolytodis	23001	CCF3566	A	Scolytodes sp. on Cecropia sp.	Costa Rica	M. Kolarik	AM267264	–	–	–
R. seticollis	R. seticollis	1031	634.66	T	Tsuga canadensis	New York, USA	R.W. Davidson	MT633076	MT629766	OM631895	OM631711
R. subalba	R. subalba	38797	121568	T	Xyleborus sp. on Persea sp.	South Carolina, USA	S. Fraedrich	–	MT629767	–	OM631712
R. subfusca	R. subfusca	38055	121571	T	Xyleborus glabratus	South Carolina, USA	S. Fraedrich	–	EU177450	–	–
R. sulcati	R. sulcati	25540	806.70	T	Gnatotrichus sulculcattus mycangium in Pseudotsuga menziesii	Canada	A. Funk	–	EU177462	–	–
R. tritirachium	R. tritirachium	25541	726.69	T	Monarthrum mali tunnel in Quercus sp.	Pennsylvania, USA	D.B. Scott	–	EU177464	–	–
R. xyleborini	R. xyleborini	45859	Hulcr6099	T	Xyleborinus andrewesii	Florida, USA	C. Bateman	MT633078	MT629769	–	–
Raffaelea sp. A (PL1001)	Raffaelea sp. A (PL1001)	38062	–		Persea sp.	California, USA	A. Eskalen	–	KJ909293	–	–
Raffaelea incertae sedis (based on our data)
R. deltoideospora	R. deltoideospora	–	WIN(M)41		Pinus sp.	Canada	J. Reid	EU879121	KT182932	–	–
Lineage X
R. vaginata	R. vaginata	40365	140086	T	Lanurgus sp. on Olea capensis	South Africa	T. Musvuugwa	KT192602	–	–	–
Sporothrix: S. candida complex
S. aemulophila	S. aemulophila	40381	140087	T	Rapanea melanophloeos	South Africa	T. Musvuugwa	OM501527	OM514854	OM631897	–
S. cabralii	S. cabralii	38098	CIEFAP456	T	Nothofagus pumilio	Argentina	A. de Errasti	OM501533	OM514861	OM631903	OM631716
S. candida	S. candida	26484	129713	T	Eucalyptus cloeziana	South Africa	G. Kamgan Nkuekam	–	OM514862	OM631904	OM631717
S. itsvo	S. itsvo	40370	141063	T	Rapanea melanophloeos	South Africa	T. Musvuugwa	KX590840	–	–	–
S. oleae #	S. oleae	40362	142082	T	Olea capensis ssp. macrocarpa wound	South Africa	T. Musvuugwa	MN298851	–	–	–
S. rapaneae	S. rapaneae	40369	141060	T	Rapanea melanophloeos	South Africa	T. Musvuugwa	OM501558	OM514885	OM631926	OM631735
Sporothrix: S. inflata complex
S. dentifunda	S. dentifunda	13016	115790	T	Quercus wood	Hungary	C. Delatour	OM501535	OM514865	OM631907	OM631718
S. dimorphospora	S. dimorphospora	12529	553.74	T	Soil	Canada	R.A.A. Morall	OM501536	OM514866	OM631908	OM631719
S. guttuliformis	S. guttuliformis	17167	437.76	T	Soil	Malaysia	T. Furukawa	OM501542	OM514873	OM631915	OM631725
S. inflata	S. inflata	12527	239.68	T	Wheat field soil	Canada	W. Gams	OM501544	–	OM631917	OM631727
Spumatoria longicollis	S. longicollis	49345	141464	E	Dung	Netherlands	J. vd Lee	–	OM514895	OM631933	–
Sporothrix: S. stenoceras & S. gossypina complexes
S. abietina *	S. abietina	22310	125.89	T	Pseudohylesinus gallery on Abies vejari	Mexico	J.G. Marmolejo	OM501526	OM514853	OM631896	OM631713
S. africana	S. africana	823	116571		Protea gaguedi	South Africa	M.J. Wingfield	OM501528	OM514855	OM631898	OM631714
S. aurorae	S. aurorae	19362	118837	T	Hylastes angustatus on Pinus elliottii	South Africa	X.D. Zhou	–	OM514857	OM631900	–
S. cantabriensis	S. cantabriensis	39766	136529	T	Hylastes attenuates on Pinus sylvestris	Spain	P. Romón	KF951554	–	–	–
S. cracoviensis #	S. cracoviensis	–	147942	T	Adult of Tryopodendron domesticum beetle on Fagus sylvaticum	Poland	R. Jankowiak	MW768964	–	–	–
S. eucastaneae	S. eucastaneae	1124	424.77	T	Canker on Castanea dentata	North Carolina, USA	R.W. Davidson	OM501539	OM514868	OM631910	OM631720
S. euskadiensis	S. euskadiensis	27318	122138	T	Hylurgops palliatus on Pinus radiata	Spain	X.D. Zhou	DQ674369	–	–	–
S. fraxini #	S. fraxini	–	147936	T	Gallery of Hylesinus varius on Fraxinus excelsior	Poland	R. Jankowiak	MH283150	–	–	–
S. fusiformis	S. fusiformis	9968	112912	T	Populus nigra	Azerbaijan	D. Aghayeva	–	OM514870	OM631912	OM631722
S. gossypina	S. gossypina	1116	ATCC18999	T	Pinus ponderosa	New Mexico, USA	R.W. Davidson	–	–	OM631914	OM631724
S. villosa #	S. villosa	–	SNM188	T	Pinus thunbergii	China	R. Chang	MW989428	–	MZ853078	–
S. lunata	S. lunata	10563	112927	T	Carpinus betulus	Austria	T. Kirisits	OM501545	–	OM631918	OM631728
S. narcissi	S. narcissi	22311	138.50	T	Narcissus sp.	Netherlands	D.P. Limber	OM501548	OM514876	OM631920	–
S. nsini	S. nsini	28602	143281	T	Protea caffra	South Africa	F. Roets	EU660458	–	–	–
S. prolifera	S. prolifera	37435	251.88	T	Quercus robur	Poland	T. Kowalski	OM501555	–	OM631924	OM631732
S. protearum	S. protearum	1107	116654		Protea caffra	South Africa	M.J. Wingfield	OM501557	OM514884	OM631925	OM631734
S. resoviensis #	S. resoviensis	–	147927	T	Wound on Betula pendula	Poland	R. Jankowiak	MH740962	–	–	–
S. rossii	S. rossii	1118	116.78	T	Dendroctonus adjunctus gallery on Pinus ponderosa	New Mexico, USA	R.W. Davidson	OM501559	OM514886	OM631927	OM631736
S. splendens *	S. splendens	23050	138722		Oodinychus sp. on Protea repens	South Africa	F. Roets	OM501561	OM514888	OM631929	OM631738
S. stenoceras	S. stenoceras	3202	237.32	T	Pine pulp	Norway	H. Robak	AF484462	DQ294350	–	–
S. uta	S. uta	40316	141069	P	Rapanea melanophloeos	South Africa	T. Musvuugwa	OM501565	OM514892	OM631930	OM631739
S. variecibata	S. variecibata	23051	121961	T	Trichouropoda sp. from Protea repens	South Africa	F. Roets	OM501566	OM514893	OM631931	OM631740
S. zambiensis	S. zambiensis	29077	124914	T	Protea caffra	Zambia	F. Roets	OM501567	OM514894	OM631932	OM631741
Sporothrix: S. pallida complex
S. albicans (syn S. pallida)	S. albicans (syn S. pallida)	17203	302.73	T	Soil	England, UK	S.B. Saksena	OM501529	OM514856	OM631899	OM631715
S. chilensis	S. chilensis	49343	139890	T	Soil	Chile	R. Cruz Choappa	–	OM514863	OM631905	–
S. gemella	S. gemella	23057	121959	T	Tarsonemus sp. on Protea caffra	South Africa	F. Roets	DQ821560	DQ821531	–	–
S. humicola	S. humicola	7618	118129	T	Soil	South Africa	H.F. Vismer	OM501543	–	OM631916	OM631726
S. mexicana	S. mexicana	29129	120341	T	Soil	Mexico	A. Espinosa	OM501547	OM514875	OM631919	–
S. pallida	S. pallida	17209	131.56	T	Stemonitis fusca	Japan	K. Tubaki	OM501550	OM514878	OM631921	OM631729
S. palmiculminata	S. palmiculminata	20677	119590	T	Protea repens	South Africa	F. Roets	OM501551	OM514879	OM631922	OM631730
S. protea-sedis	S. protea-sedis	29074	124911	P	Protea caffra	Zambia	F. Roets	OM501556	OM514883	–	OM631733
S. stylites	S. stylites	14543	118848	T	Pine utility poles	South Africa	E.M. de Meyer	OM501563	OM514890	–	–
Sporothrix: Pathogenic clade
S. brasiliensis	S. brasiliensis	29127	120339	T	Human skin	Brazil	M.D.S. Lazera	OM501531	OM514859	OM631901
S. globosa	S. globosa	29128	120340	T	Human face	Spain	C. Rubio	OM501541	OM514872	OM631913	OM631723
S. luriei	S. luriei	17210	937.72	T	Human skin	South Africa	L. Lurie	OM501546	–	–	–
S. schenckii*	S. schenckii	38850	138723		Clinical isolate	South Africa	H.F. Vismer	OM501560	OM514887	OM631928	OM631737
Sporothrix: Group D
S. cavum #	S. cavum	–	147943	T	Cavity of Dendrocopos major on Salix fragilis	Poland	R. Jankowiak	MF782813	MF782813	–	–
S. polyporicola	S. polyporicola	5461	669.88	T	Fomitopsis pinicola	Sweden	S. Ryman	OM501553	OM514881	–	–
Sporothrix: Group E
S. phasma*	S. phasma	20676	119722	T	Protea laurifolia	South Africa	F. Roets	OM501552	OM514880	OM631923	OM631731
Sporothrix: Group F
S. bragantina	S. bragantina	17149	474.91	T	Soil	Brazil	W. Gams	OM501530	OM514858	–	–
S. curviconia	S. curviconia	17164	959.73	T	Terminalia ivorensis	Ivory Coast	J. Devois	OM501534	OM514864	OM631906	–
S. epigloea	S. epigloea	22308	573.63	T	Tremella fuciformis	Argentina	R. T. Guerrero	KX590817	KX590854	–	–
S. eucalyptigena	S. eucalyptigena	45431	139899	T	Eucalyptus sp.	Australia	P. A. Barber	OM501538	–	OM631909	–
S. nebularis	S. nebularis	27319	122135	T	Hylastes attenuatus on Pinus radiata	Spain	P. Romón	KX590823	KX590862	–	–
S. nigrograna	S. nigrograna	14487	MAFF410943	T	Pinus densiflora	Japan	H. Masuya	OM501549	OM514877	–	–
S. smangaliso	S. smangaliso	50502	143341	T	Protea gaguedi	South Africa	N.P. Ngubane	MF103773	–	–	–
S. thermara	S. thermara	38930	139747	T	Cyrtogenius africus on Euphorbia ingens	South Africa	J.A. van der Linde	OM501564	OM514891	–	–
S. zhejiangensis	S. zhejiangensis	–	MUCL55183	T	Monochamus sp. on Pinus sp.	China	Q. Lu, Y.Y. Lun	KY094071	–	–	–
Sporothrix: Group G
S. dombeyi	S. dombeyi	1023	455.83	T	Nothofagus sp.	Chile	H. Butin	OM501537	OM514867	–	–
Sporothrix incertae sedis (based on our data)
Lineage XVI
S. fumea	S. fumea	26813	129712	T	Phoracantha sp. galleries on Eucalyptus	South Africa	C. Perez	OM501540	OM514869	OM631911	OM631721
Lineage XIX
S. brunneoviolacea	S. brunneoviolacea	37442	124560	P	Soil	Spain	C. Silvera	OM501532	OM514860	OM631902	–
Sporothrix incertae sedis (based on published data)
S. cryptarchum #	S. cryptarchum	–	147934	T	Adult of Cryptarcha undata	Poland	R. Jankowiak	MW768966	–	–	–
S. hypoxyli #	S. hypoxyli	47441	141569	T	Hypoxylon petriniae on Fraxinus wood	Netherlands	E. Osieck & W.J. Nel	MT637058	MW012948	–	–
S. undulata #	S. undulata	–	147929	T	Adult of Epuraea guttata	Poland	R. Jankowiak	MH740976	–	–	–
Ophiostomatales insertae sedis (based on published data)
L. antibioticum	L. antibioticum	2777	DAOM84338	T	Pinus taeda	Georgia, USA	S. Alexander	AF343677	–	–	–
L. brachiatum	L. brachiatum	2855	C388	T	Picea rubens	New York, USA	S. Alexander	AF343676	–	–	–
L. elegans	L. elegans	2245	115241		Chamaecyparis / Hinoke	Taiwan	M.J. Wingfield	AF343675	–	–	–
O. crenulatum	O. crenulatum	–	WIN(M)70-17	T	Pinus banksiana	Canada	J. Reid	–	AF135589	–	–
O. fasciatum	O. fasciatum	–	UM56		Pseudotsuga menziesii	Canada	A. Olchowecki	EU913720	EU913680	–	–
O. breviusculum	O. breviusculum	–	YCC522		Single ascospore isolate from YCC-494	Japan	Y. Yamaoka	AB200423	–	–	–
O. ssiori	O. ssiori	–	MAFF410973	T	Polygraphus ssiori on Prunus sp.	Japan	H. Masuya	AB096209	–	–	–
O. subalpinum	O. subalpinum	–	YCC408		Cryphalus sp. on Abies sp.	Japan	Y. Yamaoka	AB200424	–	LC090750	–
O. pseudonigrum	O. pseudonigrum	–	WIN(M)71-13		Picea mariana	Canada	J. Reid	–	AF135577	–	–
O. pehueninum	O. pehueninum	–	142995	T	Araucaria araucana	Chile	V. Sepúlveda	MF576438	–	MF576446	–
O. tremulo-aureum	O. tremulo-aureum	–	361.65		Black canker on Populus tremuloides	Colorado, USA	R.W. Davidson	–	AF135573	–	–

ATCC = American Type Culture Collection, Maryland, USA; CBS = the culture collection of Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands; CFCC = China Forestry Culture Collection Center, Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing, China; CIEFAP = the culture collection of the Centro de Investigación y Extensión Forestal Andino Patagónico, Argentina; CMW = the culture collection of the Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, South Africa; CTK = the culture collection of the Institute of Forest Entomology, Forest Pathology and Forest Protection, Department of Forest and Soil Sciences, University of Natural Resources and Applied Life Sciences, Vienna, Austria; CXY = the culture collection of the Chinese Academy of Forestry, China; DAOM = Canadian National Mycological Herbarium, Ottawa, Canada; KACC = Korean Agricultural Culture Collection, Suwon, Korea; KUC = Korea University Culture Collection, Seoul, Korea; MAFF = the culture collection of National Institute of Agrobiological Resources, Japan; MUCL = BCCM/MUCL Agro-food & Environmental Fungal Collection, Université catholique de Louvain, Belgium; PPRI = National Collection of Fungi, Pretoria, South Africa; RWD = the private collection of R.W. Davidson; SNM = the microbial culture collection of Shandong Normal University, Jinan, Shandong, China; TRTC = Royal Ontario Museum Fungarium, Toronto, Canada; UAMH = University of Alberta Mold Herbarium and Culture Collection, Edmonton, Canada; VPRI = Victorian Plant Pathology Herbarium, Victoria, Australia; WIN(M) = University of Manitoba, Microbiology and Botany (J. Reid’s personal collection); YCC = the private collection of Y. Yamaoka.

² T = ex-type, E = ex-epitype, P = ex-paratype; L = ex-lectotype; A = authentic isolate, used in the original study.

^* Genome data used in this study.

^# Species published after 2018, thus not included in the current analyses.

^ CMW culture replaced with a different fungus.

³ Sequences with accession numbers preceded with “OM” were generated in the current study.

Afroraffaelea (not included in the phylogenetic analyses)

Afroraffaelea C.C. Bateman et al., Fungal Ecol. 25: 46. 2017. MycoBank MB 816236. Fig. 6A, B.

Fig. 6.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Afroraffaelea. C, D. Aureovirgo. E, F. Ceratocystiopsis (a. Cop. collifera; b. Cop. ochracea; c. Cop. manitobensis; d. Cop. concentrica; e. Cop. rollhanseniana). G, H. Chrysosphaeria. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: ‘Prefix Afro - indicating its likely African origin, and ‘raffaelea’ to recognise the ecological similarity to the closely related (though probably not monophyletic) ambrosia fungus genus Raffaelea’ (Bateman et al. 2017).

Sexual morph: Unknown.

Asexual morph: No reproductive structures observed. Colonies white becoming brown with age, aerial hyphae present, abundant, subhyaline, extensively branched often perpendicularly, thickened and melanized intercalarily, distal hyphae denticulate, submerged hyphae subhyaline, constricted at septa, frequently branched, with tapering distal ends, monillioid mycelia occurring in peptone-containing medium, fragmented unicellularly or bicellularly.

Type species: Afroraffaelea ambrosiae C.C. Bateman et al., Fungal Ecol. 25: 46. 2017. MycoBank MB 816237.

No other species known.

Notes: Afroraffaelea was described by Bateman et al. (2017) and has a unique morphology relative to the rest of the Ophiostomatales. Within a few days of sub-culturing, Afr. ambrosiae (the only species in this genus) produces masses of aerial hyphae, unlike any other ophiostomatalean species, and changes colour from white to brown as the culture matures. The fungus is the dominant fungal symbiont of the ambrosia beetle Premnobius cavipennis, presumably introduced into North America from Africa (Bateman et al. 2017). The phylogenies of Bateman et al. (2017) showed Afr. ambrosiae grouping close to Fragosphaeria, but there was insufficient phylogenetic support for these authors to determine the taxonomic placement of Afroraffaelea in the Ophiostomatales.

The taxonomic placement of Afroraffaelea in our phylogenetic trees is not conclusive. The four known isolates of Afr. ambrosiae consistently grouped together as one species. However, the clade grouped inconsistently in the single gene trees. For example, in the LSU and TEF-1α trees, Afr. ambrosiae isolates grouped in Lineage VIII, but with Lineage V in ITS, and with Lineage XXIII in RPBII (data not shown). We consequently excluded these isolates from our datasets. We believe that the taxonomic placement of this fungus will be resolved when additional species are found, or possibly with whole genome data. Until such time, we treat Afroraffaelea as incertae sedis in the Ophiostomatales.

Aureovirgo (Lineage IX)

Aureovirgo J.A. van der Linde et al., Antonie van Leeuwenhoek 109: 593. 2016. MycoBank MB 813870. Fig. 6C, D.

Etymology: ‘Refers to the golden appearance of the immature ascomata and the pure white colour of the cultures (‘‘Aureovirgo’’ refers to a golden maiden with an unstated overtone of virginal whiteness)’ (Van der Linde et al. 2016).

Sexual morph: Ascomatal bases subglobose to globose, pale brown when young, becoming darker with age; necks cylindrical, dark brown. Ostiolar hyphae hyaline, parallel. Ascospores hyaline, 1-celled, enclosed in a sheath, falcate, endospores (the body of ascospores) allantoid.

Asexual morph: Leptographium-like; conidiophores macronema-tous, mononematous, hyaline; conidiogenous cells cylindrical, hyaline; conidia hyaline, 1-celled, oblong to ellipsoidal.

Type species: Aureovirgo volantis J.A. van der Linde et al., Antonie van Leeuwenhoek 109: 593. 2016. MycoBank MB 813872.

No other species known.

Notes: Aureovirgo was described from galleries of the ambrosia beetle Cyrtogenius africus in dead and dying Euphorbia ingens trees in South Africa. It is a monotypic genus accommodating A. volantis (Van der Linde et al. 2016). The ascomatal bases are honey-coloured when immature, while the rest of the culture is white. This is the only genus in the Ophiostomatales associated with Eurphorbia trees. In our dataset, Aureovirgo formed a well-supported, distinct lineage.

Ceratocystiopsis (Lineage VII)

Ceratocystiopsis H.P. Upadhyay & W.B. Kendr., Mycologia 67: 799. 1975. MycoBank MB 889. Fig. 6E, F.

Etymology: ‘Resembling the genus Ceratocystis’ (Upadhyay & Kendrick 1975, Upadhyay 1981). At the time of the description, Upadhyay & Kendrick (1975) and others treated Ophiostoma as a synonym of Ceratocystis in the Ophiostomatales. Ceratocystis was later shown to form part of the Microascales, and Ceratocystiopsis to be related to Ophiostoma in the Ophiostomatales (Zipfel et al. 2006). Although somewhat of a misnomer, the name Ceratocystiopsis remains valid and represents a distinct genus in the Ophiostomatales.

Synonym: Hyalorhinocladiella H.P. Upadhyay & W.B. Kendr., Mycologia 67: 800. 1975. MycoBank MB 8582. [Type species Hyalorhinocladiella minuta-bicolor (R.W. Davidson) H.P. Upadhyay & W.B. Kendr.]

Sexual morph: Ascomatal bases subglobose, globose, obpyriform, black or pale brown, upper part surrounded with collar-like structure or corona of globose cells; necks short conical to elongate, cylindrical, black or paler than base. Ostiolar hyphae absent or present, convergent, parallel, divergent. Asci evanescent, clavate, broadly fusiform, 8-spored. Ascospores hyaline, 1-celled, enclosed in a sheath, falcate in side view, fusiform, acicular in face view; endospores elongate orange segment-shaped in side view, cylindrical in face view; cucullate, sometimes forming bulbous swelling toward one end.

Asexual morph: Hyalorhinocladiella-like; conidiophores micronema-tous, semimacronematous, macronematous, mononematous, hyaline; conidiogenous cells integrate or discrete, polyblastic, sympodial, with denticle scars; conidia hyaline, 1-celled, obovoid, ellipsoidal, oval to globose, clavate, T-shaped, Y-shaped, cylindrical.

Type species: Ceratocystiopsis minuta (Siemaszko) H.P. Upadhyay & W.B. Kendr., Mycologia 67: 800. 1975. MycoBank MB 310480.

Other species: Listed in Table 1.

Notes: In their revision of the genus Ceratocystis, Upadhyay & Kendrick (1975) erected Ceratocystiopsis for species with falcate ascospores and a Hyalorhinocladiella asexual morph. Hausner et al. (1993a) reduced Ceratocystiopsis to synonymy with Ophiostoma based on rDNA sequences, but Zipfel et al. (2006) included additional taxa in their study and reinstated the genus that is typified by Ceratocystiopsis minuta. Species in this genus have elongated sickle-shaped (falcate) ascospores that are sheathed, produced in very short necked ascomata, characters not seen in any other genus in the Ophiostomatales (Upadhyay & Kendrick 1975, Zipfel et al. 2006, De Beer & Wingfield 2013). Species of Ceratocystiopsis are found in the galleries of conifer-infesting bark beetles (Dendroctonus species in North America or Ips species in Japan and Europe) in the Northern Hemisphere (Plattner et al. 2009).

Ceratocystiopsis spp. formed a well-supported monophyletic clade in all our phylogenies, supporting the decision of Zipfel et al. (2006) to reinstate the genus. Our data also supported the placement of Cop. neglecta (previously Ophiostoma neglectum) in Ceratocystiopsis, as shown by De Beer & Wingfield (2013).

Chrysosphaeria

Chrysosphaeria W.J. Nel et al., Mycologia 113: 1206. 2021. MycoBank MB 837564. Fig. 6G, H.

Etymology: ‘From Latin chryso-, golden, and -sphaera, sphere or orb, referring to the light colour of the ascoma bases in the type species’ (Nel et al. 2021).

Sexual morph: Ascomatal bases globose, light to golden brown; necks light brown, cylindrical, tapering towards apex, flexible. Ostiolar hyphae present, slightly divergent, hyaline. Ascospores hyaline, 1-celled, no sheath, short cylindrical to bean-shaped.

Asexual morph: Sporothrix-like; conidiophores micronematous, mononematous, hyaline; conidiogenous cells hyaline, denticulate; conidia hyaline, 1-celled, oblong, occasionally producing secondary spores hyaline, 1-celled, obovoid.

Type species: Chrysosphaeria jan-nelii W.J. Nel et al., Mycologia 113: 1206. 2021. MycoBank MB 837566.

Notes: Unlike most species in the Ophiostomatales, which occur on wood and have associations with bark or ambrosia beetles, Chyrysosphaeria was isolated from abandoned combs of the fungus-growing termite Macrotermes natalensis.

Dryadomyces (Lineage V)

Dryadomyces Gebhardt, Mycol. Res. 109: 693. 2005. MycoBank MB 28937, emend. Z.W. de Beer & M. Procter Fig 7A, B.

Fig. 7.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Dryadomyces. C, D. Esteya. E, F. Fragosphaeria. G, H. Graphilbum. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: ‘dryads, the tree nymphs in Greek mythology; referring to the habitat of these fungi in woody plants’ (Gebhardt et al. 2005).

Sexual morph: Unknown.

Asexual morph: Colony confluent, mucilaginous. Conidiophores single or aggregated in sporodochia, macronematous, mononematous, hyaline, smooth. Conidiogenous cells monillioid or oblong, sympodial, with denticles or inconspicuous scars. Conidia hyaline to subhyaline, 1-celled, globose to subglobose, obovoid to pyriform with truncate base, smooth, producing secondary spores. Aleuriospores absent or present, hyaline, globose to subglobose, terminal.

Type species: Dryadomyces amasae Gebhardt, Mycol. Res. 109: 693. 2005. MycoBank MB 369332.

Notes: Based on our analyses, Dryadomyces now includes five ambrosia beetle-associated species, previously described in either Ambrosiella, Raffaelea or Dryadomyces (Gebhardt et al. 2005, De Beer & Wingfield 2013). Raffaelea sulphurea was isolated from hardwood-infesting ambrosia beetles, as was its sister species, R. montetyi (Massoumi Alamouti et al. 2009). Two Asian species in this complex, R. quercivora from Japan and R. quercus-mongolicae from Korea, have been implicated in contributing to the death of large numbers of Quercus spp. in their native ranges (Dreaden et al. 2014).

The fifth species in Clade V of our analyses (Fig. 5), Drydomyces amasae, was initially described as the type species of the monotypic genus Dryadomyces. It was isolated from the ambrosia beetle Amasae concitatus in Taiwan (Gebhardt et al. 2005). Although the conidia of D. amasae differ from other Raffaelea species, Harrington et al. (2008) suggested that all ambrosial fungi in the Ophiostomatales should be treated in Raffaelea, until further studies revealed the taxonomic identities of these fungi. The more robust phylogenies constructed by Massoumi Alamouti et al. (2009) revealed a monophyletic lineage including D. amasae, R. sulphurea and R. montetyi. However, Harrington et al. (2010) treated D. amasae in Raffaelea, not considering the differences between conidia of D. amasae and other Raffaelea species as sufficient to separate Dryadomyces from Raffaelea.

De Beer & Wingfield (2013) named the lineage containing R. amasae, R. montetyi, R. suphurea, R. quercus-mongolicae and R. quercivora as the R. sulphurea complex, referring to the oldest of the five described species. This complex grouped within Leptographium s.l. in their phylogenies. They did, however, note that the R. sulphurea complex is most probably not part of the larger Leptographium s.l., but their data were insufficient to show otherwise. They suggested that when more robust phylogenies support the R. sulphurea complex as a monophyletic lineage separate from other genera, Dryadomyces would be a suitable name as the complex includes D. amasae.

Dreaden et al. (2014) investigated the monophyly of Raffaelea and showed that the R. sulphurea complex grouped close to isolates of E. vermicola within Leptographium s.l. as defined by De Beer & Wingfield (2013). They concluded that a larger study including additional Leptographium and Raffaelea species should provide clarity on the placement of these outlying Raffaelea lineages.

The R. sulphurea complex consistently formed a well-defined lineage distinct from Leptographium in our phylogenies. This supported re-instating the name Dryadomyces for this lineage. The description of Dryadomyces by Gebhardt et al. (2005) is emended here to accommodate other species in this lineage.

New combinations:

1) Dryadomyces montetyi (M. Morelet) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840313.

Basionym: Raffaelea montetyi M. Morelet, Ann. Soc. Sci. Nat. Archéol. Toulon Var 50: 189. 1998. MycoBank MB 445315.

Description: Morelet (1998: 189–191, fig. A).

Phylogenetic data: Gebhardt et al. (2005), Massoumi Alamouti et al. (2009), Harrington et al. (2010), Matsuda et al. (2010), De Beer & Wingfield (2013), Dreaden et al. (2014), Musvuugwa et al. (2015), De Beer et al. (2016a, b), Vanderpool et al. (2017), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: It forms part of Dryadomyces (Fig. 5), previously designated as the R. sulphurea complex (De Beer & Wingfield 2013).

2) Dryadomyces quercivorus (Kubono & Shin. Ito) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840316.

Basionym: Raffaelea quercivora Kubono & Shin. lto, Mycoscience 43: 256. 2002. MycoBank MB 483997.

Description: Kubono & lto (2002: 256–259, figs 1–11).

Phylogenetic data: Kim et al. (2009), Seo et al. (2010), Matsuda et. al. (2010), Endoh et al. (2011), Dreaden et al. (2014), Musvuugwa et al. (2015), Simmons et al. (2016), Van der Linde et al. (2016), De Beer et al. (2016a, b), Vanderpool et al. (2017), De Errasti et al. (2018), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: Dryadomyces quercivorus groups in Dryadomyces and was previously accommodated in the R. sulphurea complex (Fig. 5; De Beer & Wingfield 2013, De Errasti et al. 2018, Li et al. 2018, Saucedo-Carabez et al. 2018).

3) Dryadomyces quercus-mongolicae (K.H. Kim et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840318.

Basionym: Raffaelea quercus-mongolicae K.H. Kim et al., Mycotaxon 110: 193. 2009. MycoBank MB 515072.

Description: Kim et al. (2009: 193–195, fig. 2).

Phylogenetic data: Kim et al. (2009), Seo et al. (2010), De Beer et al. (2016a), Vanderpool et al. (2017), De Errasti et al. (2018), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: Dryadomyces quercus-mongolicae groups in Dryadomyces, and was formerly in the R. sulphurea complex (Fig. 5; De Beer & Wingfield 2013, Li et al. 2018, Saucedo-Carabez et al. 2018).

4) Dryadomyces sulphureus (L.R. Batra) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840321.

Basionym: Ambrosiella sulphurea L.R. Batra, Mycologia 59: 992. 1967. MycoBank MB 326145.

Synonyms: Raffaelea sulphurea (L.R. Batra) T.C. Harr., Mycotaxon 111: 353. 2010. MycoBank MB 515298.

Cephalosporium luteum Verrall, J. Agric. Res. 66: 141. 1943. MycoBank MB 284848.

Description: Batra (1967: 992–998, figs 20, 21, 26–29), Verrall (1943: 141, 142. fig 4).

Phylogenetic data: Cassar & Blackwell (1996), Rollins et al. (2001), Gebhardt et al. (2005), Massoumi Alamouti et al. (2009), Harrington et al. (2010), Matsuda et al. (2010), De Beer & Wingfield (2013), Dreaden et al. (2014), Musvuugwa et al. (2015), Simmons et al. (2016), De Beer et al. (2016a, b), Vanderpool et al. (2017), Saucedo-Carabez et al. (2018).

Notes: This taxon groups in Dryadomyces (Fig. 5), previously designated as the R. sulphurea species complex (De Beer & Wingfield 2013), and has a heterotypic synonym, Cephalosporium luteum (Verrall 1943). Gharabigloozare (2015) proposed C. luteum as conspecific with R. sulphurea. These two species were isolated from Xyleborinus saxeseni (syn. Xyleborus pecanis) and their galleries. The culture morphologies and conidial dimensions of Batra (1967) and Verrall (1943) match and we support their conspecificity. In terms of nomenclatural priority, Cephalosporium luteum (1943) has precedence over A. sulphurea (1967) and the former epithet has a priority over the latter as a basionym. However, since De Beer & Wingfield (2013) introduced R. sulphurea complex, the name R. sulphurea has been widely used and recognised, and we have chosen not to introduce a new epithet for this species.

Esteya (Lineage IV)

Esteya J.Y. Liou et al., Mycol. Res. 103: 243. 1999. MycoBank MB 28256, emend. Z.W. de Beer & M. Procter Fig. 7C, D.

Etymology: ‘Named in honour of Prof. Ralph H. Estey (Macdonald College, McGill University, Canada), in recognition of his contribution to the study of nematophagous fungi and nematology’ (Liou et al. 1999).

Sexual morph: Unknown.

Asexual morph: Conidiophores macronematous, mononematous, simple, subhyaline to pigmented, smooth to verrucous. Conidiogenous cells flask-shaped. Conidia hyaline, 1-celled, smooth, asymmetrically ellipsoidal in face view, concave, lunate in side view, ends moderately apiculate, with a layer of adhesive mucus on concave surface, containing ovoid endospore-like structures. Hyalorhinocladiella-like; conidiophores macronematous, mononematous, simple or branched, subhyaline to pigmented, smooth to verrucous; conidiogenous cells cylindrical to subulate; conidia hyaline, 1-celled, cylindrical, smooth.

Type species: Esteya vermicola J.Y. Liou et al., Mycol. Res. 103: 243. 1999. MycoBank MB 450702.

No other species known.

Notes: The genus Esteya was first described by Liou et al. (1999) and includes the single species, E. vermicola (Wang et al. 2015). Esteya vermicola produces two different types of conidia depending on abiotic and biotic factors, with one of the forms lethal to nematodes when consumed (Liou et al. 1999). This species was first isolated from the pine wood nematode Bursaphelencus xylophilus. It is the only nematode-associated fungus in the Ophiostomatales, clearly distinguishing Esteya from other genera in the Order. Wang et al. (2008) discovered another isolate of E. vermicola, and upon performing a BLAST search in GenBank found that members of the Ophiostomatales were the closest phylogenetic relatives. They subsequently performed molecular phylogenetic analyses including various Ophiostomatales species and found that Esteya grouped close to but separately from a clade containing two Leptographium and two Grosmannia species, and distinct from any other genus in the Ophiostomatales. The phylogenies of De Beer & Wingfield (2013) did not resolve the generic placement of Esteya in the Ophiostomatales, showing that Esteya grouped within Leptographium s.l. based on both ITS and LSU sequence data. These authors chose not to reduce Esteya to synonymy with Leptographium and recommended that Esteya should not be treated in Leptographium but be retained as a distinct genus until further studies could be conducted on the group. Later studies using LSU data again placed Esteya in Leptographium s.l. (Musvuugwa et al. 2015, De Beer et al. 2016a). Dreaden et al. (2014) used SSU, LSU and β-tub genes, which also placed Esteya in Leptographium s.l. In the phylogenies of the TEF-1α and β-tub genes generated by Wang et al. (2014), the separate treatment of Esteya from Leptographium s.l. was well supported.

In our datasets, Esteya grouped within what was previously known as Leptographium s.l. The separation of Grosmannia and Leptographium based on our data, allows the recognition of Esteya as a distinct genus, forming a well-supported clade including two of the known E. vermicola isolates. Our data did not include the type specimen of E. vermicola, because cultures of this fungus were not available in the culture collections from which isolates were sought. It is interesting that a nematode-associated fungus groups within a family of fungi not associated with nematodes. The discovery of other Esteya isolates may shed light on the evolution of this unique lifestyle within the Ophiostomatales.

Fragosphaeria (Lineage XXI)

Fragosphaeria Shear, Mycologia 15: 124. 1923. MycoBank MB 2011, emend. Z.W. de Beer & M. Procter Fig. 7E, F.

Etymology: Name is derived from the word “fragor” meaning “breaking to pieces” and “sphaera” (Shear 1923), referring to the spherical ascomata of the two species in the genus that easily break into fragments when handled.

Sexual morph: Ascomatal bases dark, globose, cleistothecial, walls easily fragmented. Asci subglobose. Ascospores yellowish-brown, 1-celled, broadly bean-shaped.

Asexual morph: Conidia hyaline, 1-celled, oblong to ellipsoidal, subalantoid, inequilateral.

Type species: Fragosphaeria purpurea Shear, Mycologia 15: 124. 1923. MycoBank MB 275760.

Other species: Listed in Table 1.

Notes: Fragosphaeria was first associated with the Ophiostomatales when a study by Suh & Blackwell (1999) showed that F. purpurea, the type species of this genus, grouped with some Ophiostoma species. The genus currently includes only two species, F. purpurea (Shear 1923) and F. reniformis (Saccardo 1881). The latter species was originally described in Cephalotheca, which Chesters (1935) reduced to synonymy with Fragosphaeria based on the similarities between F. purpurea, Cephalotheca sulfurea and C. reniformis. Malloch & Cain (1970) reinstated Fragosphaeria, transferring Cephalotheca reniformis to Fragosphaeria.

Both Fragosphaeria species are associated with stained wood within and surrounding bark beetle galleries in hardwoods and have been isolated in Britain and North America (Chesters 1935, De Beer & Wingfield 2013). Drawings by Chesters (1935) show a sporothrix-like asexual morph and allantoid ascospores produced by the cephalothecoid ascomata, which link this genus with other ophiostomatalean genera. However, the spherical cephalothecoid ascomata distinguish it from other Ophiostomatales (De Beer & Wingfield 2013). A study by Yaguchi et al. (2006) showed that there was no evident relationship between species of Cephalotheca and Fragosphaeria and confirmed that the two Fragosphaeria isolates group together. Our data confirm the treatment of Fragosphaeria as a distinct, monophyletic genus in the Ophiostomatales.

Graphilbum (Lineage VIII)

Graphilbum H.P. Upadhyay & W.B. Kendr., Mycologia 67: 800. 1975. MycoBank MB 8393, emend. Z.W. de Beer et al. In The Ophiostomatoid Fungi: Expanding Frontiers: 268. 2013. Fig. 7G, H.

Etymology: Graphilbum was described as the hyaline analogue of the genus Graphium (Upadhyay & Kendrick 1975). Graphium was later shown to form part of the Microascales (Okada et al. 1998), while Graphilbum resides in the Ophiostomatales (Zipfel et al. 2006). Although somewhat of a misnomer, the name Graphilbum remains valid and represents a distinct genus in the Ophiostomatales.

Sexual morph: Ascomatal bases globose, subglobose, black; necks dark brown to black, nearly cylindrical, straight or slightly curved. Ostiolar hyphae present or absent, parallel or divergent. Asci evanescent, broadly clavate to subglobose. Ascospores hyaline, 1-celled, cylindrical to oblong in side view, globose in end view, enclosed in rectangular sheath.

Asexual morph: Conidiophores micronematous, semimacrone-matous, macronematous, mononematous, synnematous, branched, unbranched. Hyalorhinocladiella-like; conidiophores simple or sparingly branched; conidiogenous cells sympodial, subulate; conidia hyaline, 1-celled, cylindrical or clavate to broadly clavate, oblong or ellipsoidal, sometimes slightly curved. Pesotum-like; stipes pale to dark pigmented, biverticillate; conidiogenous cells sympodial.

Type species: Graphilbum sparsum H.P. Upadhyay & W.B. Kendr., Mycologia 67: 800. 1975. MycoBank MB 314730.

Other species: Listed in Table 1.

Notes: Graphilbum was originally described to accommodate the asexual morph of some Ophiostoma (then Ceratocystis) species (Upadhyay & Kendrick 1975). Okada et al. (1998) treated Graphilbum together with other genera in the Ophiostomatales, which have synnematous asexual morphs, as a synonym of Pesotum. However, De Beer et al. (2013a) showed that the type species of Pesotum, O. ulmi, grouped within Ophiostoma s.s., while the type species of Graphilbum, O. sparsum, formed a lineage distinct from Ophiostoma s.s. They consequently reduced Pesotum to synonymy with Ophiostoma, and reinstated Graphilbum with Gra. sparsum as the type species, including seven additional species in the genus. Graphilbum species are characterised by Type E ascospores (Fig. 3H) and either a hyalorhinocladiella- or pesotum-like asexual morph (Fig. 3A, C; sensu De Beer & Wingfield 2013). This is with the exception of Gra. tsugae that produces a continuum of hyalorhinocladiella-like and pesotum-like asexual morphs. Graphilbum species are commonly found in galleries of conifer-infesting bark beetles in the Northern Hemisphere (De Beer et al. 2013b, Romón et al. 2014a, Reid & Hausner 2015, Jankowiak et al. 2020).

In our analyses, Graphilbum formed a well-supported monophyletic lineage, distinct from other genera. Interestingly, Ophiostoma pusillum consistently grouped with Graphilbum, supporting previous suggestions that the species might form part of this genus (De Beer & Wingfield 2013).

New combination:

1) Graphilbum pusillum (Masuya) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840324.

Basionym: Ophiostoma pusillum Masuya, Mycoscience 44: 302. 2003. MycoBank MB 489291.

Description: Masuya et al. (2003: 302, figs 1–10).

Phylogenetic data: Fig. 5.

Notes: Masuya et al. (2013) treated this species as part of the O. ips complex based on morphology. However, due to morphological similarities with the fungus known as Ophiostoma nigrum and Ceratocystis tubicollis (Masuya et al. 2003), De Beer et al. (2013b) suggested that it may rather belong in Graphilbum. Both O. nigrum (now Gra. nigrum) and C. tubicollis (now Gra. tubicolle) were treated in Graphilbum by De Beer et al. (2013b), and our sequence data also confirm the placement of O. pusillum in this genus (Fig. 5). The name should not be confused with S. pusilla U. Braun & Crous [= Quambalaria pusilla (U. Braun & Crous) J.A. Simpson] (De Beer et al. 2006) or Graphium pusillum (Wallr.) Sacc. (De Beer et al. 2013b).

Grosmannia (Lineage II)

Grosmannia Goid., Boll. Staz. Patol. Veg. Roma 16: 27. 1936. MycoBank MB 2141, emend. Z.W. de Beer & M. Procter. Fig. 8A–D.

Fig. 8.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Grosmannia grandifoliae complex. C, D. Grosmannia penicillata complex. E, F. Harringtonia. G, H. Hawksworthiomyces. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: Goidànich (1935) named the genus after the German forest pathologist, Helene Grosmann, later Francke-Grosmann (1900–1990), who described both the asexual morph (as Leptographium penicillatum Grosmann) and the sexual morph (as Ceratostomella penicillata Grosmann) of the type species of this genus.

Synonym: Verticicladiella S. Hughes, Canad. J. Bot. 31: 653. 1953. MycoBank MB 10394. [Type species V. abietina (Peck) S. Hughes].

Sexual morph: Ascomatal bases subglobose to globose, black; necks nearly cylindrical, straight or slightly curved, black. Ostiolar hyphae absent or present, divergent or variable. Asci evanescent. Ascospores hyaline, 1-celled, enclosed in sheath or not, endospores orange segment-shaped in side view, cylindrical in face view, globose to ellipsoidal in end view.

Asexual morph: Conidiophores macronematous, micronematous, mononematous, synnematous, singly or in groups. Hyalorhinocladiella-like; conidia hyaline, 1-celled, oblong to obovoid. Leptographium-like; conidiophores branched; stipes pale brown, branches pale brown, becoming hyaline towards apex, bases simple or rhizoid-like; conidiogenous cells hyaline; conidia hyaline, 1-celled, oblong, broadly ellipsoidal to slightly obovoid, sometimes allantoid, narrowly obovoid to clavate, straight or distinctly curved. Pesotum-like; conidiophores single or in groups; conidia hyaline, oblong, 1-celled. Sporothrix-like; conidiophores simple; conidiogenous cells hyaline, cylindrical, sympodial, denticulate; conidia hyaline, 1-celled, oblong, ovoid to ellipsoidal, clavate, sometimes developing into larger ramoconidia, producing secondary spores. Ramoconidia hyaline, 1-celled, clavate. Single ascospore cultures yeasty, budding hyaline conidia, ovoid to elongate, colony later darkened. Aleuriospores hyaline or pigmented, globose to subglobose, oval to ellipsoid.

Type species: Grosmannia penicillata (Grosmann) Goid., Boll. Staz. Patol. Veg. Roma 16: 46. 1936. MycoBank MB 253870.]

Notes: Grosmannia was originally erected by Goidànich (1935) to accommodate sexual species of Ceratostomella with leptographium-like asexual morphs (Davidson 1942). The genus was later treated as synonym of both Ophiostoma (Siemaszko 1939) and Ceratocystis (Bakshi 1951). Zipfel et al. (2006) showed that Ophiostoma and Grosmannia were distinct from each other based on ITS and LSU sequences, and separated the two genera based on these and morphological differences. However, the focus of the Zipfel et al. (2006) study was primarily on sexually reproducing species.

De Beer & Wingfield (2013) included sequence data for many more asexual Leptographium spp. in their study and showed that Leptographium and Grosmannia spp. grouped together, along with other previously unassociated Ophiostoma spp. They applied the older name, Leptographium to this group, rather than Grosmannia, which following the dual nomenclature system (McNeill et al. 2012) had preference because it was considered a sexual genus (De Beer & Wingfield 2013). They referred to the lineage as Leptographium s.l., even though the lineage did not show strong monophyletic support. The type species of Grosmannia (G. penicillata) grouped in a lineage distinct from the type species of Leptographium (L. lundbergii). However, they recommended that novel species grouping in what they referred to as the G. penicillata complex should be treated as Grosmannia species. This was until more robust analyses could confirm whether the G. penicillata complex should be treated as a distinct genus.

Based on the data emerging from the present study, we have reinstated Grosmannia as a genus distinct from Leptographium, for species that produce leptographium-like asexual morphs and have allantoid, hyaline and aseptate ascospores. These species are commonly associated with conifer-infesting bark beetles in the Northern Hemisphere (Jacobs & Wingfield 2001, Linnakoski et al. 2012). Based on our phylogenetic analyses, Grosmannia includes the fungus previously known as G. penicillata s.s. (referred to here as the G. penicillata complex), G. abieticola and L. taigense (Lineage C), and the newly recognised G. grandifoliae complex (Jankowiak et al. 2017).

Species complexes:

The G. penicillata complex

The G. penicillata complex as defined by Six et al. (2011), Linnakoski et al. (2012) and De Beer & Wingfield (2013) included 18 species. Our results and those emerging from other studies add another seven species to the complex within a well-supported lineage. All 25 species are associated in some way with conifer-infesting bark and ambrosia beetles in the Northern Hemisphere. The complex is named based on the first species in the complex to be described, G. penicillata [as Ceratostomella penicillata (Grosmann 1932)], which is also the type species of Grosmannia.

The G. grandifoliae complex

The second species complex included in our definition of Grosmannia, the G. grandifoliae complex, emerged when two new species of Leptographium were described from Poland (Jankowiak et al. 2017). Both species formed a well-supported lineage with G. grandifoliae and L. pruni, sister to the G. penicillata complex. Prior to the description of the new species, L. pruni and G. grandifoliae did not form part of the G. penicillata complex or any of the other species complexes recognised by De Beer et al. (2013b) in Leptographium s.l. Jankowiak et al. (2017) suggested that G. grandifoliae should be the name-bearing species of this small complex, because it is the oldest known name among the species that it accommodates.

Grosmannia grandifoliae is the only species for which a sexual morph has been observed and that is characterised by ascomata with very long necks (Jacobs & Wingfield 2001). With the addition of the two new species from Poland, L. trypodendri and L. betulae, it became evident that this complex represents a well-supported hardwood-infesting lineage in Grosmannia (Fig. 5) (Jankowiak et al. 2017). Leptographium trypodendri was isolated mostly from Trypodendron (beetle) species and L. betulae in association with Scolytus species (Jankowiak et al. 2017). This is the first species complex in Leptographium and Grosmannia that accommodates only hardwood-infesting species (Jankowiak et al. 2017).

In our datasets, the G. grandifoliae complex grouped within Lineage II in all except the TEF-1α tree (Fig. S3) where it grouped with Group C (below) among species complexes in Leptographium. The clear ecological distinction and the well-supported monophyletic lineage of the G. grandifoliae complex, supports its treatment as novel genus. In the future, this will need to be verified using more robust phylogenies including additional gene regions, and if possible, inclusion of a greater number of species in the complex.

Group C

In the phylogenies of Linnakoski et al. (2012), L. taigense grouped in a distinct lineage between the G. grandifoliae and G. penicillata complexes. In the LSU tree generated by De Beer & Wingfield (2013), L. taigense also grouped outside of the G. penicillata complex, close to, but not in a well-supported lineage with G. abieticola. With the additional gene regions included in this study, L. taigense and G. abieticola emerged together in a well-supported lineage distinct from, but between the two complexes defined above. Both these species are associated with conifers, as is true for all species in the G. penicillata complex. For the present, we have chosen to treat them in Grosmannia, but inclusion of newly discovered species in this lineage in the future could justify the description of a new genus to accommodate these species.

New combinations:

1) Grosmannia abieticolens (K. Jacobs & M.J. Wingf.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840331.

Basionym: Leptographium abieticolens K. Jacobs & M.J. Wingf., Mycoscience 41: 599. 2000. MycoBank MB 466545.

Description: Jacobs & Wingfield (2001: 46–48, figs 19–21).

Phylogenetic data: Jacobs et al. (2001b), Kim et al. (2004, 2005a), Masuya et al. (2004), Massoumi Alamouti et al. (2006), Paciura et al. (2010), Six et al. (2011), Duong et al. (2012), De Beer & Wingfield (2013), Huang & Chen (2014), Musvuugwa et al. (2015), Chang et al. (2019).

Notes: Sexual morph unknown. While it was not included in our study, G. abieticolens groups in the G. penicillata complex based on previously published data (Six et al. 2011, De Beer & Wingfield 2013, Chang et al. 2019). The name should not be confused with Grosmannia abieticola that formed part of Group C in our analyses.

Notes on Grosmannia abietina: Sexual morph unknown. Grosmannia abietina groups in the G. penicillata species complex (Fig. 4; Six et al. 2011, Linnakoski et al. 2012, De Beer & Wingfield 2013). The species should not be confused with Sporothrix abietina, that resides in Sporothrix. Jacobs et al. (1998) proposed the synonymy of G. engelmannii (as L. engelmannii) with G. abietina (as L. abietinum), which was supported by Jacobs & Wingfield (2001). In our analyses, an isolate of L. engelmannii (CMW 759) grouped with G. abietina in the TEF1-α tree (Fig. S3) and separate from that species in the LSU tree (Fig. S1).

2) Grosmannia altior (Paciura, Z.W. de Beer & M.J. Wingf.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840369.

Basionym: Leptographium altius Paciura et al., Persoonia 25: 106. 2010. MycoBank MB 516740.

Description: Paciura et al. (2010: 106, fig. 7h–m).

Phylogenetic data: Paciura et al. (2010), Duong et al. (2012), Linnakoski et al. (2012), De Beer & Wingfield (2013), Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. This species forms part of the G. penicillata complex based on previously published data (Paciura et al. 2010, Linnakoski et al. 2012, De Beer & Wingfield 2013, Liu et al. 2017, Chang et al. 2019).

3) Grosmannia betulae (Jankowiak et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840370.

Basionym: Leptographium betulae Jankowiak et al., Antonie van Leeuwenhoek 110: 1550. 2017. MycoBank MB 821670.

Description: Jankowiak et al. (2017: 1550, fig. 6).

Phylogenetic data: Jankowiak et al. (2017, 2018).

Notes: Sexual morph unknown. Groups in the G. grandifoliae species complex (Fig. 5; Jankowiak et al. 2017).

4) Grosmannia curviconidia (Paciura et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840373.

Basionym: Leptographium curviconidium Paciura et al., Persoonia 25: 104. 2010. MycoBank MB 516739.

Description: Paciura et al. (2010: 104–105, figs 7a–g).

Phylogenetic data: Paciura et al. (2010), Duong et al. (2012), Linnakoski et al. (2012), De Beer & Wingfield (2013), Huang & Chen (2014), Musvuugwa et al. (2015), Jankowiak et al. (2017, 2018), Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. Grosmannia curviconidia groups in the G. penicillata species complex (Fig. 5; Linnakoski et al. 2012, De Beer & Wingfield 2013, Huang & Chen 2014, Musvuugwa et al. 2015, Jankowiak et al. 2017, 2018, Liu et al. 2017, Chang et al. 2019).

5) Grosmannia euphyes (K. Jacobs & M.J. Wingf.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840376.

Basionym: Leptographium euphyes K. Jacobs & M.J. Wingf., Mycol. Res. 105: 497. 2001. MycoBank MB 467761.

Descriptions: Jacobs et al. (2001c: 496–498, figs 15–21), Jacobs & Wingfield (2001: 96–99, figs 70–72).

Phylogenetic data: Jacobs et al. (2001a), Kim et al. (2004, 2005a), Masuya et al. (2004), Massoumi Alamouti et al. (2006), Paciura et al. (2010), Six et al. (2011), Duong et al. (2012), De Beer & Wingfield (2013), Jankowiak et al. (2017, 2018), Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped in the G. penicillata complex based on previous data (Six et al. 2011, De Beer & Wingfield 2013, Jankowiak et al. 2017, 2018, Liu et al. 2017, Chang et al. 2019).

6) Grosmannia fenglinhensis (R. Chang et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840377.

Basionym: Leptographium fenglinhense R. Chang et al., Persoonia 42: 67. 2018 (2019). MycoBank MB 825092.

Description: Chang et al. (2019: 66–67, fig. 21).

Phylogenetic data: Chang et al. (2019)

Notes: Sexual morph unknown. Groups in the G. penicillata species complex (Fig. 5; Chang et al. 2019).

7) Grosmannia gestamen (Errasti & Z.W. de Beer) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840381.

Basionym: Leptographium gestamen Errasti & Z.W. de Beer, Mycol. Prog. 15: 11. 2016. MycoBank MB 814177.

Descriptions: De Errasti et al. (2016: 11–13, fig. 7).

Phylogenetic data: De Errasti et al. (2016), Jankowiak et al. (2017, 2018), Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped close to G. taigensis (as L. taigense) in the phylogenies of Jankowiak et al. (2017) and Chang et al. (2019), and with G. abieticola (Jankowiak et al. 2018). We have consequently included it in Grosmannia, as part of the complex defined by the genus in group C (Fig. 5).

8) Grosmannia innermongolica (X.W. Liu et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840384.

Basionym: Leptographium innermongolicum X.W. Liu et al., Mycol. Prog. 16: 8. 2017. MycoBank MB 811204.

Description: Liu et al. (2017: 8–10, fig. 6).

Phylogenetic data: Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped with L. taigense (Group C in Fig. 5) in the phylogenies of Liu et al. (2017) and Chang et al. (2019). It is consequently included in Grosmannia (Fig. 5).

9) Grosmannia pistaciae (Paciura et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840385.

Basionym: Leptographium pistaciae Paciura et al., Persoonia 25: 104. 2010. MycoBank MB 516738.

Description: Paciura et al. (2010: 104, figs 6g–l).

Phylogenetic data: Paciura et al. (2010), Duong et al. (2012), Linnakoski et al. (2012), De Beer & Wingfield (2013), Jankowiak et al. (2017, 2018), Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. Leptographium pistaciae has previously been shown to reside in the G. penicillata complex (Paciura et al. 2010, Linnakoski et al. 2012, De Beer & Wingfield 2013, Jankowiak et al. 2017, 2018, Liu et al. 2017, Chang et al. 2019).

10) Grosmannia pruni (Masuya & M.J. Wingf.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840386.

Basionym: Leptographium pruni Masuya & M.J. Wingf., Mycologia 96: 553. 2004. MycoBank MB 488574.

Description: Masuya et al. (2004: 553–555, figs 1–16).

Phylogenetic data: Masuya et al. (2004, 2013), Massoumi Alamouti et al. (2006), Matsuda et al. (2010), Duong et al. (2012), De Beer & Wingfield (2013), De Errasti et al. (2016), Jankowiak et al. 2017, 2018), Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped with G. grandifoliae and two other species to form the G. grandifoliae species complex (Jankowiak et al. 2017).

11) Grosmannia taigensis (Linnak. et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840387.

Basionym: Leptographium taigense Linnak. et al., Antonie van Leeuwenhoek 102: 387. 2012. MycoBank MB 564881.

Description: Linnakoski et al. (2012: 387–388, fig. 7).

Phylogenetic data: Linnakoski et al. (2012), De Beer & Wingfield (2013), Jankowiak et al. (2017, 2018), Liu et al. (2017), Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped with G. abieticola, separated from other species complexes in Grosmannia (Fig. 5), based on LSU (Fig. S1) and TEF1-α (Fig. S3) sequence data.

12) Grosmannia trypodendri (Jankowiak et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840388.

Basionym: Leptographium trypodendri Jankowiak et al., Antonie van Leeuwenhoek 110: 1546. 2017. MycoBank MB 821669.

Description: Jankowiak et al. (2017: 1546–1550, fig. 5).

Phylogenetic data: Jankowiak et al. (2017, 2018).

Notes: Sexual morph unknown. Groups in the G. grandifoliae species complex (Fig. 5).

Other species: Listed in Table 1.

Harringtonia (Lineage XI)

Harringtonia Z.W. de Beer & Procter, gen. nov. MycoBank MB 840400. Fig. 8E, F.

Etymology: Named for the American mycologist Prof. Thomas C. Harrington, who described the type species of this genus as Raffaelea lauricola (Harrington et al. 2008), and who has contributed substantially to our understanding of the biology and taxonomy of the Ophiostomatales (Harrington 1981, 1988, Harrington & Cobb 1987, Harrington et al. 2001, 2008, 2010, 2011).

Synonym: Raffaelea lauricola complex sensu Z.W. de Beer & M.J. Wingf., In The Ophiostomatoid Fungi: Expanding Frontiers: 34. 2013.

Sexual morph: Unknown.

Asexual morph: Colonies initially yeast-like at centre becoming cottony in 2 wk (Har. lauricola), cream-coloured, having submerged mycelium at margins, aging to dark green on MEA (Har. aguacate). Colonies when repeatedly subcultured forming a filamentous mycelium, with surface becoming finely tomentose (Har. brunnea). Conidiophores single or aggregated in sporodochia, hyaline, macronematous, semimacronematous, mononematous, simple or branched. Conidiogenous cells with inconspicuous annellations at point of conidial dehiscence. Conidia hyaline, 1-celled, oblong to ellipsoidal, obovoid, truncated at base, occasionally borne sessile, in some species producing secondary budding cells. Associated with ambrosia beetles.

Type species: Harringtonia lauricola (T.C. Harr. et al.) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840401.

Basionym: Raffaelea lauricola T.C. Harr. et al., Mycotaxon 104: 401. 2008. MycoBank MB 511590.

Description: Fraedrich et al. (2008: 219–220, fig. 5).

Phylogenetic data: Fraedrich et al. (2008), Harrington et al. (2008, 2010, 2011), Kim et al. (2009), Massoumi Alamouti et al. (2009), Matsuda et al. (2010), De Beer & Wingfield (2013), Dreaden et al. (2014), Musvuugwa et al. (2015), Simmons et al. (2016), De Beer et al. (2016a, b), Bateman et al. (2017), Vanderpool et al. (2018), Wingfield et al. (2017), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: Harringtonia lauricola consistently forms a clade together with Har. brunnea and Har. aguacate, that has previously been referred to as the R. lauricola complex (De Beer & Wingfield 2013). This complex has always formed a monophyletic lineage close to, but distinct from Raffaelea s.s. in previous studies (Massoumi Alamouti et al. 2009, De Beer & Wingfield 2013, Dreaden et al. 2014, Simmons et al. 2016). When De Beer & Wingfield (2013) first defined this species complex, they chose R. lauricola as the name bearing species, because it is the best-known species in this complex. This is despite the fact that it was not the first species in the complex to be described. They suggested that this lineage would likely emerge as a new genus when more robust phylogenetic analyses were performed, as is the case in the present study.

In the analyses of our datasets, the R. lauricola complex grouped close to Raffaelea s.s. and together they formed a monophyletic clade, although this relationship was not suppoted. Phylogenomic analyses conducted in the current study and previous studies (Nel et al. 2021; Vanderpool et al. 2018) showed that the R. lauricola complex and Raffaelea s.s. did not form a monophyletic lineage. Based on these results, we have removed the R. lauricola complex from Raffaelea and have described the complex as a new genus. While fully cogniscient of the fact that describing a new genus to accommodate an important tree pathogen might result in some early discomfort, this change is based on robust taxonomic data, not only from the present study. Failing to make this change perpetuates a taxonomically confused situation. In our view this decision will also provide opportunities to better understand the biology and evolutionary characteristics of two phylogenetically distinct and important groups of fungi.

All species in the R. lauricola complex produce raffaelea-like asexual morphs (De Beer & Wingfield 2013). Harringtonia lauricola is the causal agent of laurel wilt, a vascular wilt disease affecting Lauraceae species in the south-eastern USA. It is believed to have been introduced into the USA from Asia in the mycangia of Xyleborus glabratus, an ambrosia beetle invasive to the USA (Harrington et al. 2008). The other species in this complex are Har. brunnea, associated with ambrosia beetles infesting hardwoods (De Beer & Wingfield 2013), and Har. aguacate, originally isolated from avocado (Persea americana) (Simmons et al. 2016) and later shown to be vectored by X. bispinatus (Saucedo-Carabez et al. 2018).

Other new combinations:

1) Harringtonia aguacate (D.R. Simmons et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840402.

Basionym: Raffaelea aguacate D.R. Simmons et al., IMA Fungus 7: 269. 2016. MycoBank MB 817170.

Description: Simmons et al. (2016: 269, fig. 2).

Phylogenetic data: Simmons et al. (2016), Vanderpool et al. (2018), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: This species forms part of Harringtonia (Fig. 5), previously the R. lauricola complex (De Beer & Wingfield 2013).

2) Harringtonia brunnea (L.R. Batra) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840403.

Basionym: Ambrosiella brunnea L.R. Batra, Mycologia 59: 980. 1968 (1967). MycoBank MB 326140.

Synonyms: Monilia brunnea Verrall, J. Agric. Res. 66: 142. 1943. MycoBank MB 440994. nom. illegit., Art. 53. 1.

Raffaelea brunnea (L.R. Batra) T.C. Harr., Mycotaxon 111: 351. 2010. MycoBank MB 515296.

Descriptions: Verrall (1943: 142–143, fig. 5), Batra (1967: 1004–1007, figs 43, 45, 46).

Phylogenetic data: Cassar & Blackwell (1996), Rollins et al. (2001), Gebhardt et al. (2005), Massoumi Alamouti et al. (2009), Harrington et al. (2010), Matsuda et al. (2010), De Beer & Wingfield (2013), Dreaden et al. (2014), Musvuugwa et al. (2015), De Beer et al. (2016a, b), Vanderpool et al. (2018), Wingfield et al. (2017), Li et al. (2018), Saucedo-Carabez et al. (2018).

Notes: This species forms part of Harringtonia (Fig. 5), previously the R. lauricola species complex (De Beer & Wingfield 2013). This species should not be confused with Monilia brunnea, a soil inhabiting species.

Hawksworthiomyces (Lineage XXII)

Hawksworthiomyces Z.W. de Beer et al., Fungal Biol. 120: 1329. 2016. MycoBank MB 815685. Fig. 8G, H.

Etymology: ‘Named for Dr David Hawksworth, in recognition of the leading role that he has played in guiding the global mycological community through the controversial and often challenging transition from a dual nomenclature to a One Fungus-One Name-based system’ (De Beer et al. 2016b).

Sexual morph: Unknown.

Asexual morph: Conidiophores hyaline, micronematous to macronematous, mononematous, simple or branched. Conidiogenous cells hyaline, polyblastic, denticulate. Conidia hyaline, 1-celled, broadly ellipsoidal to cylindrical, producing secondary cells.

Type species: Hawksworthiomyces lignivorus (De Mey. et al.) Z.W. de Beer et al., Fungal Biol. 120: 1329. 2016. MycoBank MB 815686.

Other species: Listed in Table 1.

Notes: Hawksworthiomyces was erected to accommodate Sporothrix lignivora (now Haw. lignivora) and three other species not previously associated with the Ophiostomatales. De Beer & Wingfield (2013) listed S. lignivora as “incertae sedis”, as this species formed a lineage on its own, separate from other species in Ophiostoma and Sporothrix, even though its morphology resembled species of Sporothrix (De Meyer et al. 2008). BLAST searches on GenBank later revealed sequences from environmental studies and isolates from diversity studies with strong similarity to S. lignivora. Consequently, De Beer et al. (2016) proceeded to erect the new genus, Hawksworthiomyces, to incorporate S. lignivora and these “new” species. In the analyses of our data, Haw. lignivora, Haw. taylorii, Haw. hibbettii and Haw. crousii formed a well-supported monophyletic lineage, supporting their treatment in a distinct genus. An ENAS (Environmental Nucleic Acid Sequence) species, named Haw. sequentia ENAS, was also included in Hawksworthiomyces by De Beer et al. (2016). Hawksworthiomyces species have been isolated from diverse sources (e.g. Eucalyptus wood poles, insect fungal gardens, rhizospheres of plants) in South Africa, North and Central America, South Korea and Europe (De Beer et al. 2016). In the analyses of our datasets based on protein coding genes (Figs S3, S4) Hawksworthiomyces spp. formed two separate lineages. However, in the concatenated (Fig. 5) and LSU (Fig. S1) datasets, a monophyletic lineage emerged.

Heinzbutinia (Lineage XXIV)

Heinzbutinia Z.W. de Beer & M. Procter, gen. nov. MycoBank MB 840404. Fig. 9A, B.

Fig. 9.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Heinzbutinia. C, D. Intubia. E, F. Jamesreidia. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: Named for Prof. Heinz Butin (1928–2021), a German forest pathologist who described 10 new species in the Ophiostomatales between 1968 and 1990, including the type species of this novel genus.

Sexual morph: Ascomatal bases black, subglobose to globose; necks black, cylindrical, curved. Ostiolar hyphae absent. Asci elongate ovoid or clavate, evanescent. Ascospores hyaline, 1-celled, orange segment-like in side view, ellipsoidal in face view.

Asexual morph: Sporothrix-like; conidiophores micronematous, macronematous, mononematous, simple or branched, hyaline; conidiogenous cells sympodial, denticulate; conidia hyaline, 1-celled, ellipsoid, often curved, reniform, secondary conidia originating from swollen ellipsoidal conidia, oblong, straight or curved.

Type species: Heinzbutinia grandicarpa (Kowalski & Butin) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840405.

Basionym: Ceratocystis grandicarpa Kowalski & Butin, J. Phytopathol. 124: 243. 1989. MycoBank MB 134497.

Synonym: Ophiostoma grandicarpum [as ‘grandicarpa’] (Kowalski & Butin) Rulamort, Bull. Soc. Bot. Centre-Ouest 21: 511. 1990. MycoBank MB 130251.

Phylogenetic data: Villarreal et al. (2005), De Beer & Wingfield (2013), Reid & Hausner (2015), De Beer et al. (2016a, b), De Errasti et al. (2018).

Notes: The O. grandicarpum complex was described by De Beer et al. (2016b) for the monophyletic lineage formed by O. grandicarpum and O. microsporum among other genera in the Ophiostomatales (De Beer & Wingfield 2013, Reid & Hausner 2015, De Beer et al. 2016a, De Errasti et al. 2018). In our expanded phylogenies, the two species again consistently formed a well-supported lineage distinct from all other genera in the Ophiostomatales (Figs 5, S2–S4). Both species are characterised by very long (more than 1 mm) ascomatal necks (Kowalski & Butin 1989, Hunt 1956), sporothrix-like asexual morphs and reniform ascospores (Fig. 3F; Type A as categorised by De Beer & Wingfield 2013). Both species were isolated from hardwoods. Based on the distinct phylogenetic placement and supported by a unique morphology and ecology, we have designated the O. grandicarpum complex as the new genus Heinzbutinia.

Other new combinations:

1) Heinzbutinia microspora (Arx) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840406.

Basionym: Ophiostoma microsporum Arx, Antonie van Leeuwenhoek 18: 211. 1952. MycoBank MB 302079.

Synonyms: Ceratostomella microspora R.W. Davidson, Mycologia 34: 650. 1942. MycoBank MB 284860. nom. illegit., Art. 53.1, later homonym for Cs. microspora Ellis & Everh., see De Beer et al. 2013, Section C.1.

Ceratocystis perparvispora J. Hunt, Lloydia 19: 46. 1956. MycoBank MB 294227. Art. 52.1, superfluous nom. nov.

Ceratocystis microspora (R.W. Davidson) R.W. Davidson & Aoshima, Ph.D. thesis, University of Tokyo: 20. 1965. nom. inval., Arts 29.1, 39.1 or 39.2.

Ceratocystis microspora (Arx) R.W. Davidson, J. Colorado-Wyoming Acad. Sci. 6: 16. 1969. MycoBank MB 453851.

Descriptions: Hunt (1956: 46–47), Griffin (1968: 710), De Hoog (1974: 63–64, fig. 25), Olchowecki & Reid (1974: 1709), Upadhyay (1981: 50, figs 104–108), Maekawa et al. (1987: 8, 10, figs 1–6).

Phylogenetic data: Hausner et al. (1993b), Mullineux et al. (2011), De Beer & Wingfield (2013), Musvuugwa et al. (2015), De Beer et al. (2016a, b).

Notes: This species grouped with He. grandicarpa and distinct from other Ophiostoma spp. (De Beer et al. 2016a, b), in the O. grandicarpum complex as designated by De Beer et al. (2016b). See note under He. grandicarpa. The name He. microspora should not be confused with Leptographium microsporum or Ceratostomella microspora (see De Beer et al. 2013, section C.2).

2) Heinzbutinia solheimii (Strzałka & Jankowiak) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840407.

Basionym: Ophiostoma solheimii Strzałka & Jankowiak, Antonie van Leeuwenhoek 112: 1517. 2019. MycoBank MB 830198.

Description: Jankowiak et al. (2019: 1516–1517, fig. 8).

Phylogenetic data: Jankowiak et al. (2019).

Notes: This species is closely related to and formed a well-supported lineage with He. grandicarpa and He. microspora (Jandowiak et al. 2019, figs 1, S4).

Intubia

Intubia W.J. Nel et al., Mycologia 113: 1206. 2021. MycoBank MB 837565. Fig. 9C, D.

Etymology: ‘From the Xhosa language, Intubi for termite, recognising the source where the fungus was found’ (Nel et al. 2021).

Sexual morph: Ascomatal bases dark brown, globose; necks uniformly dark, often slightly curved, tapering towards apex, extremely long. Ostiolar hyphae absent. Asci not seen. Ascospores produced in slimy droplet at apex of neck, hyaline, 1-celled, cylindrical, sometimes slightly curved, no sheath.

Asexual morph: Conidiophores micronematous, hyaline, arising singly. Hyalorhinocladiella-like; conidiogenous cells smooth; conidia hyaline, 1-celled, bacilliform tapering toward one end, occasionally producing secondary spores. Sporothrix-like; conidiogenous cells denticulate; conidia of two types, first type formed on vegetative hyphae, hyaline, 1-celled, round to obovoid, second type formed on conidiophores, hyaline, 1-celled, bacilliform, secondary conidia present.

Type species: Intubia macrotermitinarum W.J. Nel et al., Mycologia 113: 1208. 2021. MycoBank MB 837567.

Other species: Listed in Table 1.

Notes: Distinguished from other phylogenetically related genera in the Ophiostomatales for their unique habitat with dark coloured ascomata embedded in the substrate of abandoned Termitomyces combs.

Jamesreidia (Lineage XII)

Jamesreidia Z.W. de Beer & M. Procter, gen. nov. MycoBank MB 840408. Fig. 9E, F.

Etymology: Named for the Canadian mycologist, Dr James Reid, who described 28 novel ophiostomatoid species, including the type species of this new genus. He was also involved in producing some of the first phylogenetic data based on ribosomal sequences distinguishing between the Ophiostomatales and Microascales (Olchowecki & Reid 1974, Hausner et al. 1992).

Synonym: Ophiostoma tenellum complex sensu Z.W. de Beer & M.J. Wingf., In The Ophiostomatoid Fungi: Expanding Frontiers: 34. 2013.

Sexual morph: Ascomatal bases black, globose; necks absent or present, black, straight or bent. Ostiolar hyphae divergent. Asci evanescent, clavate to subglobose. Ascospores hyaline, 1-celled, orange segment-like in side view, oblong in face view, broadly ellipsoidal to globose in end view.

Asexual morph: Conidiophores macronematous, semimacronema-tous, mononematous, hyaline, simple or branched. Conidiogenous cells polyblastic, denticulate. Conidia hyaline, 1-celled, broadly ovoid to nearly cylindrical, slightly curved.

Type species: Jamesreidia tenella (R.W. Davidson) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840409.

Basionym: Ceratocystis tenella R.W. Davidson, Mycologia 50: 666. 1958. MycoBank MB 294240.

Synonyms: Ophiostoma tenellum (R.W. Davidson) M. Villarreal, Mycotaxon 92: 263. 2005. MycoBank MB 346181.

Ceratocystis capitata H.D. Griffin, Canad. J. Bot. 46: 699. 1968. MycoBank MB 327623.

Descriptions: Griffin (1968: 713, 715, fig. 93, pI. Ill), Olchowecki & Reid (1974: 1708, pI. XVI figs 307, 308, 311, 312), Upadhyay (1981: 114, figs 408–412), Maekawa et al. (1987: 10–11, figs 19, 20), Hutchison & Reid (1988a: 68).

Phylogenetic data: Villarreal et al. (2005), Linnakoski et al. (2010, 2016), De Beer & Wingfield (2013), De Errasti et al. (2018), De Beer et al. (2016a), De Errasti et al. (2018), Wang et al. (2018), Chang et al. (2019).

Notes on the type species: Jamesreidia tenella groups with J. coronata, J. nigricarpa (Fig. 5; Linnakoski et al. 2010, De Beer & Wingfield 2013) and J. rostrocoronata (in the phylogenies of De Beer et al. 2016a) to form Jamesreidia, previously designated as the O. tenellum species complex (De Beer & Wingfield 2013). This species serves as the type species of the new genus, due to the fact that it is the oldest described species in the genus.

Notes on the genus: The O. tenellum complex was defined in Ophiostoma s.l. by De Beer & Wingfield (2013), who noted that members of the complex shared a similar morphology with species in the S. schenckii-O. stenoceras complex (also treated in Ophiostoma s.l. at the time, but now recognised as the genus Sporothrix). The asexual morphs of all species can be described as sporothrix-like and sexual morphs form Type A ascospores (Fig. 3F; as categorised by De Beer & Wingfield 2013). All species in this complex colonise conifer wood in North America (Linnakoski et al. 2010).

De Beer et al. (2016a) noted that the generic status of the O. tenellum complex should be further investigated, because it grouped distinctly from both Sporothrix and Ophiostoma s.s. in their phylogenies. In the present study, this complex grouped separately from what we have designated as Ophiostoma and Sporothrix in our phylogeny (Fig. 5) as well as in the phylogenies of other authors (Villarreal et al. 2005, Linnakoski et al. 2010, De Beer & Wingfield 2013, De Beer et al. 2016a). We have thus elevated the status of the O. tenellum complex to that of a genus, for which we have provided the name Jamesreidia.

Other new combinations:

1) Jamesreidia coronata (Olchow. & J. Reid) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840410.

Basionym: Ceratocystis coronata Olchow. & J. Reid, Canad. J. Bot. 52: 1705. 1974. MycoBank MB 310490.

Synonym: Ophiostoma coronatum (Olchow. & J. Reid) M. Villarreal, Mycotaxon 92: 263. 2005. MycoBank MB 346368.

Description: Hutchison & Reid (1988a: 66, 68).

Phylogenetic data: Hausner et al. (1993b), Thwaites et al. (2005), Villarreal et al. (2005), Linnakoski et al. (2010), Mullineux et al. (2011), De Beer & Wingfield (2013), Linnakoski et al. (2016), De Beer et al. (2016a), De Errasti et al. (2018), Wang et al. (2018), Chang et al. (2019).

Notes: This species grouped in Jamesreidia (Fig. 5), previously designated as the O. tenellum species complex (De Beer & Wingfield 2013).

2) Jamesreidia nigrocarpa (R.W. Davidson) M. Procter & Z.W. de Beer, comb. nov. [MycoBank MB 840411]

Basionym: Ceratocystis nigrocarpa R.W. Davidson, Mycopathol. Mycol. Appl. 28: 276. 1966. MycoBank MB 327637.

Synonym: Ophiostoma nigrocarpum (R.W. Davidson) de Hoog, Stud. Mycol. 7: 62. 1974. MycoBank MB 319024.

Descriptions: De Hoog (1974: 62–63, fig. 24), Olchowecki & Reid (1974: 1709), Upadhyay (1981: 104, figs 378–381), Benade et al. (1997: 1110–1111, figs 6–11).

Phylogenetic data: Aghayeva et al. (2004), Zhou et al. (2004a, 2006), Roets et al. (2006, 2008, 2010), Zipfel et al. (2006), De Meyer et al. (2008), Linnakoski et al. (2010), Madrid et al. (2010), Romón et al. (2014b), De Errasti et al. (2016, 2018), Linnakoski et al. (2016), De Beer et al. (2016a), Giraldo et al. (2017), Chang et al. (2019).

Notes: This species forms part of Jamesreidia (Fig. 5), previously the O. tenellum species complex (De Beer & Wingfield 2013).

3) Jamesreidia rostrocoronata (R.W. Davidson & Eslyn) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840412.

Basionym: Ceratocystis rostrocoronata R.W Davidson & Eslyn, Mem. N.Y. Bot. Gard. 28: 50. 1976. MycoBank MB 310518.

Synonym: Ophiostoma rostrocoronatum (R.W. Davidson & Eslyn) de Hoog & R.J. Scheff., Mycologia 76: 297. 1984. MycoBank MB 107077.

Descriptions: Upadhyay (1981: 112), Hutchison & Reid (1988a: 76–78).

Phylogenetic data: Hausner et al. (1993b), Jacobs et al. (2003), Villarreal et al. (2005), Linnakoski et al. (2010), De Beer & Wingfield (2013), Reid & Hausner (2015), De Beer et al. (2016b), Osorio et al. (2016), Jankowiak et al. (2017b), Wang et al. (2018).

Notes: A LSU sequence generated by Jacobs et al. (2003) was shown to group in Sporothrix (De Beer & Wingfield 2013, Osorio et al. 2016, Jankowiak et al. 2017, Wang et al. 2018). However, sequence data generated by De Beer et al. (2016b) for the same isolate, placed O. rostrocoronatum with the O. tenellum complex. Based on analyses of sequences from four gene regions and morphological characteristics, De Beer et al. (2016b) proceeded to treat O. rostrocoronatum in the O. tenellum complex. We have consequently treated this species as part of the lineage described here as Jamesreidia.

Leptographium (Lineage I)

Leptographium Lagerb. & Melin, Svenska Skogsv.-Fören. Tidskr. 25: 257. 1927. MycoBank MB 8749, emend. Z.W. de Beer & M. Procter. Figs 10, 11.

Fig. 10.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Leptographium clavigerum complex. C, D. Leptographium galeiforme complex. E, F. Leptographium lundbergii complex. G, H. Leptographium olivaceum complex. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Fig. 11.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Leptographium piceiperdum complex. C, D. Leptographium procerum complex. E, F. Leptographium serpens complex. G, H. Leptographium wageneri complex. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: ‘A thin, small brush. From the Greek adjective, λɛπτóζ (thin) and the Greek noun γραϕισν (a small brush). The generic name refers to the conidiophores that resemble small brushes’ (Jacobs & Wingfield 2001).

Synonyms: Scopularia Preuss, Linnaea 24: 133. 1851. MycoBank MB 22345, nom. illegit., Art. 53.1. (Type species Sc. venusta Preuss).

Phialographium H.P. Upadhyay & W.B. Kendr., Mycologia 66: 183. 1974. MycoBank MB 9340. (Type species Ph. sagmatosporae H.P. Upadhyay & W.B. Kendr.).

Graphiocladiella H.P. Upadhyay, A Monograph of Ceratocystis and Ceratocystiopsis: 138. 1981. MycoBank MB 8394. (Type species G. clavigera H.P. Upadhyay).

?= Europhium A.K. Parker, Canad. J. Bot. 35: 175. 1957. MycoBank MB 1938. [Type species E. trinacriforme A.K. Parker; Placement uncertain, see De Beer et al. (2013b)].

Sexual morph: Ascomata cleitothecial or perithecial; bases black, subglobose to globose; necks absent or present, very short, elongated, cylindrical, straight or flexuous. Ostiolar hyphae absent. Asci evanescent, clavate to broadly clavate. Ascospores hyaline, 1-celled, enclosed in sheath, reniform or cucullate in side view, oblong or rectangular in face view, ovoid, globose or triangular in end view.

Asexual morph: Conidiophores macronematous, micronematous, mononematous, singly or in groups, upright or prone, branches brown, becoming paler towards upper branches, bases simple or rhizoid-like. Conidiogenous cells hyaline, cylindrical. Conidia hyaline, 1-celled, oblong, pyriform, broadly ellipsoidal to obovoid, with truncate base. Hyalorhinocladiella-like; conidiophores simple or branched, upright; conidia hyaline, 1-celled, oblong to ellipsoid. Pesotum-like; conidiophores pale to dark brown, hyaline to light grey, reddish brown at apex; conidia hyaline, 0–4-celled, oblong, cylindrical to clavate, broadly fusiform, or subglobose to ellipsoid with truncate base. Ramoconidia hyaline, clavate to ellipsoid, 1-celled spores. Serprentine hyphae present in L. lundbergii, L. serpens complexes and L. douglasii in L. wageneri complex.

Type species: Leptographium lundbergii Lagerb. & Melin, Svenska Skogsv.-Fören. Tidskr. 25: 248. 1927. MycoBank MB 269891.

Synonym: Scopularia lundbergii (Lagerb. & Melin) Goid., Ann. Mycol. 31: 138. 1933. MycoBank MB 253103.

Notes on the genus: Leptographium was originally described as an asexual genus with L. lundbergii as type species (Lagerberg et al. 1927). Soon afterwards, the sexual genus Grosmannia was described to accommodate the sexual morph of another species, Leptographium penicillatum (Goidànich 1935). During the course of the next 65 years, an additional 28 asexual species were described in Leptographium and 17 species with known sexual morphs that were treated in different genera (Harrington 1988, Wingfield 1993). This prompted a revision of the genus in a comprehensive monograph by Jacobs & Wingfield (2001), who treated the sexual morphs of these species in Ophiostoma, of which Grosmannia was listed as one of several synonyms (Jacobs & Wingfield 2001). As mentioned in the introduction to this study, Zipfel et al. (2006) reinstated Grosmannia for all sexual species with Leptographium asexual morphs. However, they included only one asexual species, L. lundbergii, in their phylogeny.

De Beer & Wingfield (2013) were the first authors to apply the One Fungus-One Name principles to the Ophiostomatales, and listed Grosmannia as synonym of the older name, Leptographium. However, they recognised that the type species for these two genera grouped in different lineages but concluded that their LSU and ITS phylogenies were not sufficiently robust to resolve the generic status of these sub-lineages within what they referred to as ‘Leptographium sensu lato’. They listed 94 valid species in this broad concept but refrained from making new combinations for species that did not have names in Leptographium, recommending that phylogenies based on a greater number of gene regions should be used to resolve the genera.

Prior to the study of De Beer & Wingfield (2013), several authors defined phylogenetic species complexes in ‘Leptographium sensu lato’ (Massoumi Alamouti et al. 2011, Six et al. 2011, Duong et al. 2012, Linnakoski et al. 2012). The results of the present study clearly show that ‘Leptographium sensu lato’ represents two distinct genera. These include Leptographium (Lineage I) with L. lundbergii as type species and including eight species complexes defined below and Grosmannia (Lineage II) defined by G. penicillata including two species complexes defined above. A third complex could emerge as new species are described in Grosmannia.

The last formal morphological description of Leptographium was provided by Jacobs & Wingfield (2001) for asexual fungi forming mononematous conidiophores and branched conidiogenous apparatuses. In view of the One Fungus-One Name principles, we have emended the description of Leptographium to accommodate the morphology of the sexual morphs, treated previously in Grosmannia, Ophiostoma, Ceratocystis or Europhium (Upadhyay 1981, Jacobs & Wingfield 2001, De Beer & Wingfield. 2013). Morphology of synnematous asexual morphs previously excluded from Leptographium and treated variously as species of Graphium, Pesotum, Phialographium or Graphiocladiella (Upadhyay & Kendrick 1974, Upadhyay 1981, Seifert & Okada 1993, Okada et al. 1998) have also been incorporated in the revised circumscription of the genus.

Species complexes:

The L. clavigerum complex

The L. clavigerum complex was first described by Massoumi Alamouti et al. (2011) as the G. clavigera complex. Grosmannia clavigera is also the type species of Graphiocladiella, a genus originally described for the asexual morphs of ‘Ceratocystis’ species forming mononematous and synnematous conidiophores (Upadhyay 1981, Harrington 1988). Harrington (1988) proceeded to reduce Graphiocladiella to synonymy with Graphium (in which he also included species producing leptographium-like conidiophores). Species in the G. clavigera complex produce mostly synnematous aggregates of leptographium-like conidiophores, cleistothecal ascomata and reniform ascospores with hat-shaped sheaths (Linnakoski et al. 2012, De Beer & Wingfield 2013).

The first whole genome sequence produced for species in the Ophiostomatales was that for G. clavigera. This work was justified by the close association of G. clavigera with the mountain pine beetle (Dendroctonus ponderosae), a native but invasive pest causing devastating losses in the Northwestern USA and Canada (Kim et al. 2004, Massoumi Alamouti et al. 2011, De Beer & Wingfield 2013). Genome data were then used to show that what was previously thought to be a highly variable population of G. clavigera, actually represents a complex including multiple species (Massoumi Alamouti et al. 2011). Later studies contributed additional species to this new species complex (Six et al. 2011, De Beer & Wingfield 2013). In the present study, the G. clavigera complex formed a well-supported monophyletic lineage, and currently includes eight well-defined species, as well as one undescribed species (Massoumi Alamouti et al. 2011).

The L. galeiforme complex

The L. galeiforme complex was first described as the O. galeiformis complex when Zhou et al. (2004b) designated an epitype for the species (Bakshi 1951). Kim et al. (2005b) showed that G. radiaticola was part of the complex, and that this species was conspecific with the asexual species, Hyalopesotum pini. The latter species name was thus used for the asexual morph of G. radiaticola (Kim et al. 2005b, De Beer & Wingfield 2013). However, when the One Fungus–One Name principles are applied, the oldest epithet ‘pini’ (Hutchison & Reid 1988b) should take precedence over ‘radiaticola’ (Kim et al. 2005b). We have consequently provided a new combination, L. pini, to represent this species. It was previously believed that G. radiaticola and G. galeiformis were the same species (Thwaites et al. 2005). However, data from the studies of Kim et al. (2005b) and Linnakoski et al. (2012) showed that these are distinct species, and that in addition to these, the complex includes two as yet undescribed cryptic species. Chang et al. (2017) reported L. pini (as G. radiaticola) from China, as well as another possible new species in the complex.

All species in the L. galeiforme complex produce synnemata that appear to be loose aggregates of leptographium-like conidiophores (Zhou et al. 2004b, Linnakoski et al. 2012, De Beer & Wingfield 2013). The distinction between the two species is also supported by the ecology of these fungi, with L. galeiforme being associated with conifer-infesting bark beetles in Europe, and L. pini (as G. radiaticola) associated with pine-infesting bark beetles with a wider distribution (South Africa, Chile, Europe, China, Korea, and New Zealand; Linnakoski et al. 2012).

The L. lundbergii complex

The L. lundbergii complex was first defined by Jacobs et al. (2005). In a study emerging from a survey of ophiostomatoid fungi in Finland and Russia, Linnakoski et al. (2012) expanded the complex to include additional species that group close to L. lundbergii. Although these species always grouped close together, they never formed a well-supported monophyletic lineage, neither in the analyses of Linnakoski et al. (2012), various subsequent analyses (De Beer & Wingfield 2013, Jankowiak et al. 2017, 2018, Chang et al. 2017, Pan et al. 2020) nor those in the present study. However, for the sake of convenience, we have chosen to treat these species as a group. All the species produce relatively short conidiophores that have conidia with truncate bases, and the two sexually reproducing species produce cucullate ascospores (Linnakoski et al. 2012, De Beer & Wingfield 2013). Most of the species in this complex have been described from conifers in Asia and they have also been found in Europe, Canada, South Africa and New Zealand as the causal agents of sapstain in lumber (Linnakoski et al. 2012).

The L. olivaceum complex

The L. olivaceum complex accommodates species that are characterised by brownish synnematous asexual morphs and cucullate ascospores produced in ascomata with almost cylindrical necks and prominent ostiolar hyphae (Yin et al. 2019). The complex was first demarcated in a phylogeny by Six et al. (2011), more comprehensively defined by Linnakoski et al. (2012), and subsequently recognised by De Beer & Wingfield (2013) and Jankowiak et al. (2017, 2018). The complex includes L. sagmatosporum, the type species of the genus Phialographium (Upadhyay & Kendrick 1974), and its inclusion in the complex renders that name a synonym of Leptographium (De Beer & Wingfield 2013).

Most recently, Yin et al. (2019) added six new species to the complex. All species in this complex are associated with various bark beetles, mostly infesting spruce and pine trees in North America and Eurasia (Linnakoski et al. 2012, Jankowiak et al. 2017, 2018, Yin et al. 2019). The only exception is L. francke-grosmanniae that groups peripheral to the complex, has a morphology quite distinct from other species and was isolated from a bark beetle gallery on oak trees in Europe (Davidson 1971, Mouton et al. 1992, Yin et al. 2019). It is possible that this species represents a distinct species complex that cannot be distinguished at the present time.

The L. piceiperdum complex

Linnakoski et al. (2012) first defined this species complex when they showed that isolates described as G. piceiperda separated into five distinct species, two of which were unnamed. Ando et al. (2016) focused on isolates from different sources in Japan, revealing several additional cryptic species. Both Linnakoski et al. (2012) and Ando et al. (2016) concluded that epitypification is needed for L. piceiperdum and L. europhioides in order to confirm the placement of these species, because sequences for them were derived from herbarium type material. All members of this complex form typical leptographium-like asexual morphs, have cucullate ascospores (De Beer & Wingfield 2013, Ando et al. 2016), and have been found in association with conifer infesting bark beetles (mostly Ips species) in Europe, North America, Russia and Japan (Linnakoski et al. 2012, Ando et al. 2016).

The L. procerum complex

The L. procerum complex was first defined by De Beer & Wingfield (2013), and studied by Yin et al. (2015). These authors used seven gene regions to evaluate the species boundaries in the complex, and showed that it accommodates nine species. No sexual morph has been observed for any species in this complex, and asexual morphs can be described as leptographium-like (Jacobs et al. 2000b, 2006, Jacobs & Wingfield 2001, Lu et al. 2008, Paciura et al. 2010, De Beer & Wingfield 2013, Yin et al. 2015). Species in this complex are associated with conifer-infesting bark beetles in North America and Eurasia, with most described from East Asia. The exception is L. profanum that occurs on hardwood trees in the USA (Linnakoski et al. 2012, Yin et al. 2015). Most species in this complex are not widely distributed, with single species being found only in single countries. However, L. procerum has been isolated from Pinus spp. in China, Russia and the USA (Yin et al. 2015).

The L. procerum complex has received considerable attention by researchers. This is due to the fact that L. procerum, the type species of the complex has been associated with a root and root-collar disease of Pinus spp. in the USA known as white pine root decline (Kendrick 1962, Dochinger 1967, Wingfield 1983, Wingfield et aI. 1994). Furthermore, L. procerum is associated with the red turpentine beetle (Dendroctonus valens), which is native to North America and possibly western Europe and was introduced into China resulting in devastating losses of Chinese native pines (Yan et al. 2005, Lu et al. 2008, Taerum et al. 2012, 2013, 2017, Zhou et al. 2013, Sun et al. 2013, Yin et al. 2015).

The L. serpens complex

The L. serpens species complex was first recognised based on phylogenetic analyses by Six et al. (2011). In a focused study to resolve typification issues in the complex, Duong et al. (2012) showed that L. serpens, previously recognised as a single taxon represented five distinct species. All of these species produce typical leptographium-like conidiophores, with characteristic serpentine hyphae on agar, while ascospores produced by two of the species are sheathed (Wingfield & Marasas 1980, Jacobs & Wingfield 2001, Duong et al. 2012). Interestingly, most of the members of the complex have relatively narrow distributions, most being known from only a single country. For example, L. serpens occurs in Italy, L. gibbsii in the UK, L. yamaoake in Japan and L. castellanum in Spain and the Dominican Republic. The exception here is L. alacre that is more widely distributed and associated with conifer-root infesting bark beetles in South Africa and Italy (Duong et al. 2012), although its occurrence in South Africa is certainly due to an accidental introduction likely from Europe. Marincowitz et al. (2017) described a morphologically similar species, L. rhodanense from Switzerland that groups between the L. serpens and the L. wageneri complexes.

The L. wageneri complex

Six et al. (2011) first defined the L. wageneri species complex based on phylogenetic analyses. All species in this complex form typical leptographium-like asexual morphs. However, the sexual morph has been observed only for L. wageneri var. ponderosae, and this was described as G. wageneri (Goheen & Cobb 1978, Zipfel et al. 2006). This fungus produces allantoid ascospores, but unlike other Leptographium ascospores, no sheath was reported (Goheen & Cobb 1978, Jacobs & Wingfield 2001). Most of the species in the complex are associated with conifer-root infesting beetles in the USA (Cobb et al. 1974, 1984, Hansen et al. 1988, Witcosky & Hansen 1985, Witcosky et al. 1986). The only exception is L. reconditum, which was isolated from the roots of Triticum species in South Africa (Jooste 1978).

The L. wageneri complex includes three host-specific varieties that are important pathogens of conifers restricted to the western USA and causing black stain root disease (Cobb 1988, Hessburg & Hansen 2000). These three formally described varieties can be distinguished from each other based on their morphological characteristics and the host tree species that they infect (Harrington & Cobb 1984, 1987, Jacobs & Wingfield 2001). Leptographium wageneri var. pseudotsugae occurs on Douglas fir (Pseudotsuga menziesii), L. wageneri var. ponderosae on hard pines (Pinus ponderosa, P. jeffreyi and P. contorta), and L. wageneri var. wageneri on pinyon pines (P. monophylla and P. edulis). Phylogenetic analyses in this study support the lineage defined as the G. wageneri complex by De Beer & Wingfield (2013) and Six et al. (2011).

Groups A & B

Leptographium pineti grouped singly in Leptographium and this is consistent with phylogenies emerging from other studies (Paciura et al. 2010, Six et al. 2011, Duong et al. 2012, De Beer & Wingfield 2013). Chang et al. (2017) described a new species from China, L. ningerense (not included in our data), which grouped with L. pineti in their datasets.

Grosmannia cainii formed a lineage distinct from others in Leptographium in our analyses (Fig. 5) and this was also seen in the phylogenies of De Beer & Wingfield (2013).

New combinations:

1) Leptographium cainii (Olchow. & J. Reid) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840389.

Basionym: Ceratocystis cainii Olchow. & J. Reid, Canad. J. Bot. 52: 1697. 1974. MycoBank MB 310486.

Synonyms: Ophiostoma cainii (Olchow. & J. Reid) T.C. Harr., Mycotaxon 28: 41. 1987. MycoBank MB 128912.

Grosmannia cainii (Olchow. & J. Reid) Zipfel, Z.W de Beer & M.J. Wingf., Stud. Mycol. 55: 89. 2006. MycoBank MB 500812.

Descriptions: Upadhyay (1981: 39, figs 43–47), Seifert & Okada (1993: 32, fig. 3D).

Phylogenetic data: Hausner et al. (2000), Masuya et al. (2004), Kim et al. (2005b), Six et al. (2011), Duong et al. (2012), De Beer & Wingfield (2013), Jankowiak et al. (2017, 2018), Liu et al. (2017), De Errasti et al. (2018), Chang et al. (2019).

Notes: Leptographium cainii groups alone and distinct from other species complexes in Leptographium in our data (Fig. 5). This is consistent with the data presented by De Beer & Wingfield (2013).

2) Leptographium europhioides (E.F. Wright & Cain) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840392.

Basionym: Ceratocystis europhioides E.F. Wright & Cain, Canad. J. Bot. 39: 1222. 1961. MycoBank MB 327627.

Synonyms: Ophiostoma europhioides (E.F. Wright & Cain) H. Solheim, Nordic J. Bot. 6: 203. 1986. MycoBank MB 102979.

Grosmannia europhioides (E.F. Wright & Cain) Zipfel, Z.W. de Beer & M.J. Wingf., Stud. Mycol. 55: 90. 2006. MycoBank MB 500818.

Ceratocystis shikotsuensis Aoshima, Ph. D. thesis, University of Tokyo: 10. 1965. nom. inval., Arts 29.1, 39.1 or 39.2.

Descriptions: Davidson et al. (1967: 929–930), Griffin (1968: 709, 713), Olchowecki & Reid (1974: 1699, pI. XIII, figs 259–261), De Hoog & Scheffer (1984: 295, fig. 2), Yamaoka et al. (1997: 1221–1222), Jacobs et al. (1998: 290–291), Jacobs et al. (2000a: 239).

Phylogenetic data: Hausner et al. (1993b, 2000), Okada et al. (1998), Schroeder et al. (2001), Masuya et al. (2004), Greif et al. (2006), Mullineux & Hausner (2009), Matsuda et al. (2010), Paciura et al. (2010), Six et al. (2011), Linnakoski et al. (2012), De Beer & Wingfield (2013), Musvuugwa et al. (2015), Ando et al. (2016), Yamaoka (2017), De Errasti et al. (2018).

Notes: Leptographium europhiodes groups in the L. piceiperdum complex (Fig. 5). De Beer & Wingfield (2013) noted that species such as G. europhioides, previously treated as synonyms of G. piceiperda based on morphology (Jacobs et al. 2000a, Jacobs & Wingfield 2001), were possibly distinct species (Linnakoski et al. 2012). Ando et al. (2016) investigated the phylogenetic relationships of Japanese isolates assigned to the L. piceiperdum complex and recognised 13 lineages within the complex. These included lineages representing G. aenigmatica, G. laricis, G. piceiperda group D and eight representing distinct, but undescribed species (Ando et al. 2016, Yamaoka 2017). Ando et al. (2016) concluded that although L. piceiperdum (as G. piceiperda) and L. europhioides (as G. europhioides) are valid species, epitypification is needed for both species to resolve the identity of the remaining undescribed taxa.

3) Leptographium galeiforme (B.K. Bakshi) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840393.

Basionym: Ceratocystis galeiformis Bakshi, Mycol. Pap. 35: 13. 1951. MycoBank MB 294208.

Synonyms: Ophiostoma galeiforme (B.K. Bakshi) Math.-Käärik, Meddeland. Statens Skogs-Forstningsinst. 43: 47. 1953. MycoBank MB 302075 (as ‘galeiformis’).

Grosmannia galeiformis (B.K. Bakshi) Zipfel, Z.W. de Beer & M.J. Wingf., Stud. Mycol. 55: 90. 2006. MycoBank MB 500820.

Descriptions: Mathiesen-Käärik (1953: 47–50), Hunt (1956: 33), Wingfield (1993: 48, fig. 8), Zhou et al. (2004b: 1309–1311, fig. 2).

Phylogenetic data: Hausner et al. (2000), Zhou et al. (2004b), Kim et al. (2005b, 2011), Thwaites et al. (2005), Greif et al. (2006), Zipfel et al. (2006), Lu et al. (2009), Mullineux & Hausner (2009), Harrington et al. (2010), Matsuda et al. (2010), Paciura et al. (2010), Six et al. (2011), Duong et al. (2012), Linnakoski et al. (2012), De Beer & Wingfield (2013), Taerum et al. (2013), Masuya et al. (2013), Huang & Chen (2014), Wang et al. (2014), Musvuugwa et al. (2015), De Beer et al. (2016a, b), Chang et al. (2017, 2019), Wingfield et al. (2017), Jankowiak et al. (2017), Liu et al. (2017), De Errasti et al. (2018).

Notes: This species grouped with L. radiaticola and undescribed species (Linnakoski et al. 2012, De Beer & Wingfield 2013) to form the L. galeiforme species complex (Fig. 5). In the phylogenies of Chang et al. (2019), L. koraiensis grouped within this species complex. The latter species was not included in our analyses.

4) Leptographium pseudoeurophioides (Olchow. & J. Reid) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840413.

Basionym: Ceratocystis pseudoeurophioides Olchow. & J. Reid, Canad. J. Bot. 52: 1700. 1974. MycoBank MB 310514.

Synonyms: Ophiostoma pseudoeurophioides (Olchow. & J. Reid) Hausner et al., Canad. J. Bot. 71: 1264. 1993. MycoBank MB 362667.

Grosmannia pseudoeurophioides (Olchow. & J. Reid) Zipfel et al., Stud. Mycol. 55: 91. 2006. MycoBank MB 500826.

Descriptions: Olchowecki & Reid (1974: 1700, figs 219–229).

Phylogenetic data: Hausner et al. (1993b, 2000), Masuya et al. (2004), Mullineux & Hausner (2009), Mullineux et al. (2011), De Beer & Wingfield (2013), Musvuugwa et al. (2015), De Errasti et al. (2018).

Notes: Leptographium pseudoeurophioides was assigned to the G. penicillata complex (Hausner et al. 1993b, 2000). However, based on a short LSU sequence produced by Hausner et al. (1993b, 2000) and ascospore morphology, De Beer & Wingfield (2013) suggested that L. pseudoeurophioides should form part of the L. piceiperdum complex, and that additional collections and fresh material would be required to confirm its placement. For the present, we treat this species in Leptographium.

5) Leptographium radiaticola (J.J. Kim et al.) M. Procter & Z.W. de Beer comb. nov. [MycoBank MB 840396]

Basionym: Ophiostoma radiaticola J.J. Kim et al., Mycotaxon 91: 486. 2005. MycoBank MB 500832.

Synonyms: Grosmannia radiaticola (J.J. Kim et al.) Z.W. de Beer & M.J. Wingf., Stud. Mycol. 55: 91. 2006. MycoBank MB 500827.

Hyalopesotum pini L.J. Hutchison & J. Reid, New Zealand J. Bot. 26: 90. 1988. MycoBank MB 135442.

= Pesotum pini (L.J. Hutchison & J. Reid) G. Okada & Seifert, Canad. J. Bot. 76: 1504. 1998. MycoBank MB 446360.

Descriptions: Hutchison & Reid (1988b: 90–91, figs 32–35, of Hy. pini), Kim et al. (2005b: 486–489, figs 1–14).

Phylogenetic data: Masuya et al. (2004), Kim et al. (2005a, b), Thwaites et al. (2005), Zipfel et al. (2006), Lu et al. (2009), Mullineux & Hausner (2009), Paciura et al. (2010), Six et al. (2011), Duong et al. (2012), Linnakoski et al. (2012), De Beer & Wingfield (2013), Taerum et al. (2013), Huang & Chen (2014), Wang et al. (2014), Romón et al. (2014a), Musvuuwga et al. (2015), Chang et al. (2017, 2019), Liu et al. (2017), De Errasti et al. (2018).

Notes: This species grouped in the L. galeiforme species complex (Fig. 5; Linnakoski et al. 2012, De Beer & Wingfield 2013). Pesotum pini has been recognised as the asexual morph of L. radiaticola (Kim et al. 2005b). Following the One Fungus-One Name principles, the oldest name, irrespective of morph, should take precedence in the selection of the basionym. In this case, Hyalopesotum pini would serve that purpose. However, to avoid confusion with the name O. pini, currently treated as a synonym of O. minus, but possibly a distinct taxon (De Beer & Wingfield, 2013), we have designated O. radiaticola as basionym.

Other species: Listed in Table 1.

Leptographium and Grosmannia incertae sedis (Lineages III & VI)

Leptographium piriforme (Lineage III)

De Beer & Wingfield (2013) applied a relatively wide taxonomic concept for Leptographium s.l. and their phylogenies placed L. piriforme in a distinct lineage with G. crassivaginata. The data in the present study did not include G. crassivaginata. Greif et al. (2006) noted some morphological similarities between L. piriforme and G. crassivaginata when they described L. piriforme. Consequently, G. crassivaginata may form part of Lineage III if it were included in analyses.

Leptographium piriforme produces curved conidia and pear-shaped cells, distinguishing it from other Leptographium species (Greif et al. 2006, Jankowiak & Kolařik 2010). In the concatenated dataset utilised in the present study, L. piriforme grouped basal to Lineage I and II (Fig. 5). Although the phylogeny presented by Greif et al. (2006) was much smaller than that presented here, L. piriforme also grouped outside of the lineage containing Leptographium and Grosmannia. For the present, we have retained L. piriforme in Leptographium although its taxonomic placement clearly deserves further consideration.

Leptographium verrucosum (Lineage VI)

Gebhardt et al. (2002) was the first to describe L. verrucosum (as O. verrucosum) and their description was based solely on morphology. De Beer & Wingfield (2013) transferred O. verrucosum to Leptographium based on LSU sequences, and their recommendation was that new species grouping with either Leptographium or Grosmannia should be described in Leptographium. In our LSU dataset (Fig. S1), L. verrucosum grouped basal to Lineage I (Leptographium s.s.), but this grouping was not seen in the other gene trees. For the present and until additional material is available for further analyses, we have chosen to retain it in Leptographium.

Masuyamyces (Lineage XVII)

Masuyamyces Z.W. de Beer & M. Procter, gen. nov. MycoBank MB 840414. Fig. 12A, B.

Fig. 12.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Masuyamyces. C, D. Ophiostoma clavatum complex. E, F. Ophiostoma ips complex. G, H. Ophiostoma minus complex. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Etymology: Named for Dr Hayato Masuya, a Japanese mycologist who has described 17 species of ophiostomatoid fungi, including the type species of this novel genus.

Sexual morph: Ascomatal bases black, subglobose to globose; necks black, nearly cylindrical, curved or straight. Ostiolar hyphae absent or present. Asci evanescent, clavate to sub-globose when young. Ascospores hyaline, 1-celled, oblong or cylindrical in front view, allantoid in side view, globose in end view, enclosed in uniform hyaline sheath.

Asexual morph: Conidiophores macronematous, mononematous to synnematous. Hyalorhinocladiella-like; conidiophores simple or branched, hyaline; conidiogenous cells annellidic, cylindrical; conidia hyaline, obovoid to globose, oblong to ellipsoidal, 1-celled. Pesotum-like; conidiophores loosely compacted, stipes hyaline to pale brown; conidiogenous cells hyaline annellidic; conidia hyaline, oblong to ellipsoidal.

Type species: Masuyamyces botuliformis (Masuya) Z.W. de Beer & M. Procter, comb. nov. MycoBank MB 840415.

Basionym: Ophiostoma botuliforme Masuya, Mycoscience 44: 304. 2003. MycoBank MB 489292.

Description: Masuya et al. (2003: 304, figs 11–21).

Phylogenetic data: Fig. 5.

Notes on the type species: No DNA sequence data were previously available for this species (De Beer & Wingfield 2013). Although M. botuliformis resembles O. allantosporum morphologically (Masuya et al. 2003), it did not group with the latter species, which groups in Ophiostoma (Fig. 5). Recently, sequences of M. botuliformis (as O. botuliforme) have been deposited in GenBank, which are identical to our ITS and LSU sequences. See note under M. saponiodorus.

Notes on the genus: In our phylogenies, Lineage XVII consistently formed a monophyletic group, distinct from Ophiostoma. This group includes O. ambrosium, O. botuliforme, O. pallidulum (not included in our analyses) and O. saponiodorum. All described from conifers and all having reniform ascospores (Fig. 3F; Type A as categorised by De Beer & Wingfield 2013). Although O. pallidulum was not included in our analyses, it grouped with O. saponiodorum in the phylogenies of Linnakoski et al. (2010) and De Beer & Wingfield (2013).

The isolate of O. ambrosium included in our analyses is not the ex-type culture but was designated as an “isolate from type collection” by Hausner et al. (1993b). Our LSU sequence is almost identical to that of Hausner et al. (1993b), although theirs is very short. However, the morphology of this isolate does not match that of the type of O. ambrosium, and although this isolate is from the type collection, it may represent a different species.

Our isolate of O. saponiodorum was also not the ex-type isolate for the species, but our ITS sequence is identical to the one produced by Linnakoski et al. (2010). The latter species grouped with isolate CMW 30883 and was used in the cross to produce the holotype of the species. Therefore, although two species in this clade are not represented by ex-type isolates, we have sufficient confidence in the sequences to justify describing this lineage as a new genus.

Masuyamyces ambrosius was described from Betula albae infested by the beetle Trypodendron domesticum in Great Britain (Bakshi 1950). Masuyamyces botuliformis was isolated from Pinus densiflora in Japan (Masuya et al. 2003), M. pallidulus was from P. sylvestris infested by Hylastes brunneus in Finland (Linnakoski et al. 2010), and M. saponiodorus was from Picea abies infested with Ips typographus and Pityogenes chalcographus in Russia as well as Finland (Linnakoski et al. 2010). The asexual morph of M. botuliformis was described as pesotum-like (Masuya et al. 2003), that of M. ambrosium as raffaelea-like (De Beer & Wingfield 2013), that of M. pallidulus as hyalorhinocladiella-like, and that of M. saponiodorus as pesotum-like with a hyalorhinocladiella-like synasexual morph (Linnakoski et al. 2010).

Other new combinations:

1) Masuyamyces acarorum (R. Chang & Z.W. de Beer) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840416.

Basionym: Ophiostoma acarorum R. Chang & Z.W. de Beer, MycoKeys 28: 40. 2017. MycoBank MB 823693.

Description: Chang et al. (2017: 40–41, fig. 7).

Phylogenetic data: Chang et al. (2017, 2019).

Notes: Sexual morph unknown. This species resides in Masuyamyces and grouped with M. saponiodorus (as O. saponiodorum) in the phylogenies of Chang et al. (2017, 2019). See note under M. saponiodorus.

2) Masuyamyces ambrosius (B.K. Bakshi) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840526.

Basionym: Ceratocystis ambrosia B.K. Bakshi, Trans. Brit. Myc. Soc. 33: 116. 1950. MycoBank MB 294192.

Synonym: Ophiostoma ambrosium (B.K. Bakshi) Georg Hausner, J. Reid & Klassen, Canad. J. Bot. 71: 1264. 1993. MycoBank MB 362662.

Description: Bakshi (1950: 116–118, fig. 2)

Notes: Sexual morph unknown.This species grouped with M. saponiodorus and M. botuliformis (Fig. 5).

3) Masuyamyces jilinensis (R. Chang et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840417.

Basionym: Ophiostoma jilinense R. Chang et al., MycoKeys 28: 63. 2019. MycoBank MB 825086.

Description: Chang et al. (2019: 63–65, fig. 16).

Phylogenetic data: Chang et al. (2019).

Notes: Sexual morph unknown. This species grouped with M. saponiodorus (as O. saponiodorum) in the phylogeny of Chang et al. (2019), and is consequently included in the the genus. See note under M. saponiodorus.

4) Masuyamyces lotiformis (Z. Wang & Q. Lu) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840418.

Basionym: Ophiostoma lotiforme Z. Wang & Q. Lu, IMA Fungus 11: 17. 2020. MycoBank MB 830612.

Description: Wang et al. (2020: 17–18, fig. 14)

Phylogenetic data: Wang et al. (2020).

Notes: Sexual morph unknown. This species grouped with M. saponiodorus (as O. saponiodorum) in the phylogeny of Wang et al. (2020), and is consequently included in the genus. See note under M. saponiodorus.

5) Masuyamyces massonianae M. Procter & Z.W. de Beer, sp. nov. MycoBank MB 840419.

Synonym: Ophiostoma massonianae H.M. Wang & Q. Lu, MycoKeys 39: 15. 2018. nom. inval., Art. 40.7. MycoBank MB 827856.

Description: Wang et al. (2018: 15–17, fig. 4).

Holotype: PREM 63310.

Phylogenetic data: Wang et al. (2018)

Notes: Sexual morph unknown. This species grouped with M. saponiodorus (as O. saponiodorum) in the phylogenies of Wang et al. (2018), and is thus included in the genus. See note under M. saponiodorus.

6) Masuyamyces pallidulus (Linnak. et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840420.

Basionym: Ophiostoma pallidulum Linnak. et al., Persoonia 25: 86. 2010. MycoBank MB 518884.

Description: Linnakoski et al. (2010: 86, 88, fig. 9).

Phylogenetic data: Linnakoski et al. (2010, 2016), De Beer & Wingfield (2013), De Beer et al. (2016a, b), Chang et al. (2017, 2019), Wang et al. (2018).

Notes: Sexual morph unknown. Although not included in our analyses, this species was shown to consistently group with M. saponiodorus (as O. saponiodorum) and is thus treated in Masuyamyces (Linnakoski et al. 2010, 2016, De Beer & Wingfield 2013, Chang et al. 2017, 2019, Wang et al. 2018). See note under M. saponiodorus.

7) Masuyamyces saponiodorus (Linnak. et al.) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 840421.

Basionym: Ophiostoma saponiodorum Linnak. et al., Persoonia 25: 88. 2010. MycoBank MB 518885.

Description: Linnakoski et al. (2010: 88, fig. 10).

Phylogenetic data: Linnakoski et al. (2010, 2016), Six et al. (2011), De Beer & Wingfield (2013), Romón et al. (2014b), Chang et al. (2017, 2019), Wang et al. (2018).

Notes: Masuyamyces saponiodorus consistently grouped separately from what we have defined as Ophiostoma in our phylogenies (Fig. 5, S1–S4), and with the other species listed above in previously published phylogenies (Linnakoski et al. 2010, 2016, De Beer & Wingfield 2013, Chang et al. 2017, 2019, Wang et al. 2018).

Ophiostoma (Lineage XV)

Ophiostoma Syd. & P. Syd., Ann. Mycol. 17: 43. 1919. MycoBank MB 3614, emend. Z.W. de Beer & M. Procter. Figs 12C–13F.

Fig. 13.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Ophiostoma piceae complex. C, D. Ophiostoma pluriannulatum complex. E, F. Ophiostoma ulmi complex. G, H. Raffaelea. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Synonyms: Linostoma Höhn., Ann. Mycol. 16: 91. 1918. MycoBank MB 2885. nom. illegit. Art. 53.1, De Beer et al. 2013a. [Type species Linostoma piliferum (Fr.) Höhn.].

Pesotum J.L. Crane & Schokn., Amer. J. Bot. 60: 347. 1973. MycoBank MB 9270. [Type species Pesotum ulmi (M.B. Schwarz) J.L. Crane & Schokn.].

Hyalopesotum H.P. Upadhyay & W.B. Kendr., Mycologia 67: 801. 1975. MycoBank MB 8579. (Type species H. introcitrinum H.P. Upadhyay & W.B. Kendr.).

Pachnodium H.P. Upadhyay & W.B. Kendr., Mycologia 67: 802. 1975. Asexual synonym: MycoBank MB 9189 (Type species P. canum H.P. Upadhyay & W.B. Kendr.).

Etymology: Ophio- from the Greek for ‘snake-like’ and ‘stoma’ meaning mouth, referring to the long, tubular necks of the ascomata.

Sexual morph: Ascomatal bases black, subglobose to globose; necks black, cylindrical, straight or slightly curved. Ostiolar hyphae absent or present, divergent. Asci evanescent, clavate. Ascospores hyaline, 1-celled, enclosed in sheath, allantoid, reniform, cylindrical to ossiform in side view, ellipsoid in face view, globose in end view.

Asexual morph: Conidiophores macronematous, micronematous, mononematous, synnematous. Hyalorhinocladiella-like; conidiophores simple or branched; conidia hyaline, 1-celled, oblong to ellipsoid. Leptographium-like; conidiophores upright or prone, bases simple or rhizoid-like; conidiogenous cells hyaline, subcylindrical; conidia hyaline, 1-celled, oblong to obovoid with truncate base. Pesotum-like; conidiophores single or in groups, black becoming paler towards apex; stipes extending beyond conidiogenous cells, becoming seta-like structures (O. cupulatum); conidia hyaline, mostly 1-celled, oblong to ellipsoid, round apex, tapering towards base, clavate to obovoid; conidia elongated cylindrical or clavate with tapering towards base, 1–3-celled (O. nikkonse), giving rise to secondary conidia (O. nikkoense). Sporothrix-like; conidiogenous cells proliferating, sympodial, denticulate; conidia hyaline, 1-celled, oblong to ellipsoid, elongate or broadly cylindrical, round apex, tapering towards base, clavate or obovoid, obpyriform, giving rise to secondary spores (O. cupulatum, O. flexuosum).

Type species: Ophiostoma piliferum (Fr.) Syd. & P. Syd., Ann. Mycol. 17: 43. 1919. MycoBank MB 431882.

Other species: Listed in Table 1.

Notes on the genus: The definition of Ophiostoma emerging from this study relates to what was previously treated as Ophiostoma s.l. by De Beer & Wingfield (2013). In this case it excludes the S. schenckii-O. stenoceras complex (now Sporothrix) and the O. tenellum complex. Species in the newly defined Ophiostoma sensu stricto typically produce Type A or B ascospores (Fig. 3F, G; as categorised by De Beer & Wingfield 2013) and a variety of asexual morphs including those that are sporothrix-like, pesotum-like, hyalorhinocladiella-like; some having more than one morph and commonly a continuum between these morphs (De Beer & Wingfield 2013).

Species complexes:

The O. clavatum complex

The O. clavatum complex was defined by Linnakoski et al. (2016). Species in this complex generally have rectangular to cylindrical shaped ascospores (Fig. 3F; Type A as categorised by De Beer & Wingfield 2013), spirally coiled ostiolar hyphae that are brown in colour and they produce pesotum-like asexual morphs (Mathiesen-Käärik 1960, Linnakoski et al. 2016). These fungi are found in Eurasia and are mostly associated with conifer-infesting bark beetles residing in the genus Ips and some species cause blue stain in the sapwood of trees (Linnakoski et al. 2016).

Ophiostoma ainoae and O. tapionis grouped peripherally to other species in the O. clavatum complex in the datasets of Linnakoski et al. (2016). Ophiostoma ainoae has ascospores similar in shape to species in the O. ips complex, but groups within the O. clavatum complex (Linnakoski et al. 2016). Ophiostoma ainoae also has a hyphal morphology similar to that of O. clavatum but it grouped peripheral to that complex (Linnakoski et al. 2016). Due to its morphological similarity with other species in the complex, O. ainoae is best treated as part of the O. clavatum species complex. This grouping was also supported in our analyses (Fig. 5).

The O. ips complex

The O. ips complex was first defined by De Beer & Wingfield (2013). Species in the complex all have cylindrical ascospores in a pillow-shaped sheath (Fig. 3H; Type E as categorised by De Beer & Wingfield 2013) and this distinguishes them from other species in Ophiostoma. These species form pesotum- to hyalorhinocladiella-like asexual morphs (De Beer & Wingfield 2013). Members of this complex are associated with bark beetles in the genera Ips and Dendroctonus, infesting conifers mostly in North America and areas where pine trees have been introduced (Upadhyay 1981, Zhou et al. 2004a, Linnakoski et al. 2010).

The ex-type isolate of the asexual species Hyalorhinocladiella ips (CMW 14175) grouped in our analyses within the O. ips complex, similar to the results of De Beer & Wingfield (2013). Hyalorhinocladiella ips was originally described as Tuberculariella ips (Leach et al. 1934), later treated in Ambrosiella (Batra 1967) and most recently in Hyalorhinocladiella (Harrington et al. 2010). This fungus has been suggested to represent the asexual morph of O. montium, because the two species share morphological similarities and there are minimal differences in their sequence data (Massoumi Alamouti et al. 2009, De Beer & Wingfield 2013, De Beer et al. 2013b). In our analyses (Fig. 5) O. montium and “H. ips” grouped very close to each other. De Beer et al. (2013b) formally listed H. ips as synonym of O. montium (Rumbold 1941), even though H. ips is the older of the two names (Leach et al. 1934) and should thus be designated as basionym for the species. However, the epithet “ips” is already in use for O. ips, the name bearing species of this complex, that has Ceratostomella ips (Rumbold 1931) as basionym. To avoid the creation of a later homonym by using H. ips as basionym, we have chosen to select Ceratostomella montium (Rumbold 1941) as basionym for the species and for which O. montium (Von Arx 1952) is then the appropriate current name. The O. ips complex formed a well-supported lineage that might be excluded from Ophiostoma s.s. in future to be treated as a distinct genus. However, for the present and in the absence of more robust datasets, we have retained species in this complex in Ophiostoma s.s.

The O. minus complex

The O. minus complex was well supported in our phylogenies even though this was not true in the phylogenies of De Beer & Wingfield (2013). The O. minus complex was first defined by Jacobs & Kirisits (2003). Species in this complex produce hyalorhinocladiella-like asexual morphs (Fig. 3A; as categorised by De Beer & Wingfield 2013) and have been isolated from conifers in North America and Europe. Ophiostoma minus is a well-known causal agent of blue stain in conifers. It is associated with the southern pine beetle (Dendroctonus frontalis), a native but invasive pest in the USA (Jacobs & Kirisits 2003, Gorton et al. 2004).

It must be noted that our phylogenies included isolates of only the North American form of O. minus and O. pseudominus, both available in the CMW collection. This was due to the fact that there were no available cultures of other species included in the complex by previous authors (Jacobs & Kirisits 2003, Gorton et al. 2004, Linnakoski et al. 2010, De Beer & Wingfield 2013).

The O. piceae complex

Harrington et al. (2001) first defined the O. piceae complex, but the complex was not applied by De Beer & Wingfield (2013). Because the complex did not have any phylogenetic support in their trees, De Beer and Wingfield (2013) treated the species in the complex as part of Ophiostoma s.s. Yin et al. (2016) redefined the O. piceae complex, including the discripiton of three new species described from spruce-infesting bark beetles in China. The two sub-clades seen to be formed by the O. piceae complex (Harrington et al. 2001) are now known as the O. ulmi complex and the O. piceae complex. The O. ulmi complex includes what was previously referred to as the ‘hardwood clade’, and the newly redefined O. piceae complex includes what was previously referred to as the ‘conifer clade’ (Harrington et al. 2001, Grobbelaar et al. 2010, 2011, Kamgan Nkuekam et al. 2011, De Beer & Wingfield 2013, Yin et al. 2016). Species in this complex form ascomata with long necks of up to 500 μm and can form either sporothrix- or pesotum-like asexual morphs, or both, while some form hyalorhinocladiella-like asexual morphs (Fig. 3A, C, E; as categorised by De Beer & Wingfield 2013). Members of this complex have been isolated from conifers mostly in the Northern Hemisphere, and one species (O. cupulatum) was described from New Zealand (Yin et al. 2016). In our phylogenies, the O. piceae complex grouped distinct from the O. ulmi complex.

The O. pluriannulatum complex

The O. pluriannulatum complex was initially defined as the O. multiannulatum complex (Villarreal et al. 2005) and later renamed to the O. pluriannulatum complex (Kamgan Nkuekam et al. 2008, Zanzot et al. 2010, De Beer & Wingfield 2013). Ascospores of species in the complex are allantoid (Fig. 3F; Type A in De Beer & Wingfield 2013) (Hunt 1956, Kamgan Nkuekam et al. 2008, Zanzot et al. 2010, De Beer & Wingfield 2013). This is with the exception of O. carpenteri that has Type B (Fig. 3G) narrowly clavate ascospores (Hausner et al. 2003, De Beer & Wingfield 2013). Other than O. carpenteri, all members of the O. pluriannulatum complex have very long ascomatal necks relative to other Ophiostoma species (Zanzot et al. 2010, De Beer & Wingfield, 2013). Ophiostoma carpenteri has very short necks but does produce a sporothrix-like asexual morph as in all other members of the complex (Hausner et al. 2003, De Beer & Wingfield 2013). Species in this complex have been found in Eurasia, North America and Africa on conifers and hardwoods, and some associated with bark beetles in the genera Ips and Dendroctonus (Zhou et al. 2004a, Linnakoski et al. 2010, Pacuira et al. 2010). This complex may represent a discrete genus, but our analyses were not sufficiently robust to support such a distinction.

The O. ulmi complex

The O. ulmi complex was first known as the ‘hardwood clade’ in the O. piceae complex (Harrington et al. 2001, Linnakoski et al. 2010). It was later referred to as the O. quercus complex (Kamgan Nkuekam et al. 2011), until De Beer & Wingfield (2013) defined it as the O. ulmi species complex. Most of the members of the complex produce allantoid ascospores (Fig. 3F; Type A) that lack sheaths (Hunt 1956, Brasier 1991, De Beer & Wingfield, 2013). Species in this complex occur in Eurasia, Australia, Africa and North and South America and are associated with a variety of bark beetles (Upadhyay 1981, Brasier 1991, Brasier & Mehrotra 1995, Villarreal et al. 2005, Harrington et al. 2001, Kamgan Nkuekam et al. 2010, 2011, Linnakoski et al. 2008, 2009, Grobbelaar et al. 2010, Paciura et al. 2010). De Beer & Wingfield (2013) chose to have O. ulmi as the name bearing species for this complex, justified because it is the best-known species in this complex, as one of the causal agents of Dutch Elm Disease (DED). Ophiostoma novo-ulmi and O. himal-ulmi are also causal agents of DED (Brasier 1991, Harrington et al. 2001). DED is one of the most catastrophic tree disease pandemics in the Northern Hemisphere, leading to the death of large populations of Elm trees (Brasier 1979, 1991, Gibbs 1978). Another species in this complex, O. quercus, is associated with substantial economic losses due to sapstain in Quercus spp. (Harrington et al. 2001).

Ophiostoma bacillisporum, O. torulosum and O. undulatum have asexual morphs that are different in morphology to other members of the O. ulmi complex. Their asexual morphs are mycelial while other members of the complex form sporothrix- and pesotum-like asexual morphs. The ascospores of the species are, however, similar to those in the O. ulmi complex (De Beer & Wingfield, 2013). We have supported the decision of De Beer & Wingfield (2013) to include species residing in what was known as the O. quercus complex (Kamgan Nkuekam et al. 2011) in the O. ulmi complex. This decision is justified based on the fact that O. quercus groups within the O. ulmi complex. However, our analyses grouped O. torulosum outside the O. piceae complex (Fig. 5) and its position requires further consideration.

Group H

De Beer & Wingfield (2013) included O. triangulosporium in the O. ulmi complex but noted that its unique ascospore morphology (allantoid ascospores with triangular sheaths) suggested that its placement in the complex required further study, including inspection of the type material and a greater number of isolates. In our analyses (Fig. 5), O. triangulosporium grouped peripheral to other species in the O. ulmi complex.

Group I

Ophiostoma macrosporum and an undescribed Ophiostoma species (previously treated as a Hyalorhinocladiella sp.) formed a small lineage outside of any of the species complexes in Ophiostoma.

Group J

The type species of Ophiostoma, O. piliferum, grouped on its own within Ophiostoma. Although it is the type of the genus, it does not group in any species complex (De Beer & Wingfield 2013, Yin et al. 2016, De Beer et al. 2016a, b).

Group K

Ophiostoma tetropii grouped peripheral to the O. minus and O. piceae complexes (Fig. 5). Linnakoski et al. (2010) included O. tetropii in the O. minus complex. However, the ascospores of O. tetropii have a distinct morphology different to species in the O. minus complex (De Beer & Wingfield 2013). The phylogenies of De Beer & Wingfield (2013) also showed that O. tetropii grouping separately from the O. minus complex and it clearly requires further study.

Ophiostoma incertae sedis (Lineages XVIII, XX & XXIII)

Ophiostoma valdivianum (Lineage XVIII)

As mentioned above, O. valdivianum formed a lineage with S. fumea in the phylogenies of De Beer et al. (2016a), grouping basal to most of Sporothrix. However, in the phylogenies arising from the present study, these two species did not group together.

Ophiostoma angusticollis and O. denticulatum (Lineage XX)

Ophiostoma angusticollis and O. denticulatum consistently formed a lineage distinct from Ophiostoma. In the phylogenies of De Beer & Wingfield (2013), O. angusticollis formed a lineage with O. sejunctum (not included in the present study) close to the O. tenellum complex (Lineage XIII) but distinct from Ophiostoma. We have chosen not to describe this lineage as a new genus because we were not able to include the holotype of O. angusticollis.

Ophiostoma noisomeae (Lineage XXIII)

Ophiostoma noisomeae consistently grouped distinct from Ophiostoma in our datasets. In the phylogenies of De Beer et al. (2016a), O. noisomeae also grouped distinct from Ophiostoma.

Paleoambrosia (No sequence data available)

Paleoambrosia Poinar & F.E. Vega, Fungal Biol. 122: 1160. 2018. MycoBank MB 840457. nom. inval., Art. F.5.1.

Etymology: ‘From the Greek “palaios” = ancient, and the Greek “ambrosia” = immortal, referring to the fossilized ambrosial fungus discovered with a Platypodine beetle in ~100 million-year-old amber in Myanmar’ (Poinar & Vega 2018).

Sexual morph: Unknown.

Asexual morph: Conidiophores borne in sporodochia, simple, black. Conidia black, 1-celled, globose to obovoid, borne in slimy droplet terminally. Yeast-like cells present in mycangium.

Type species: Paleoambrosia entomophila Poinar & F.E. Vega, Fungal Biol. 122(12): 1160 (2018). MycoBank MB 840458. nom. inval., Art. F.5.1.

No other species known at present.

Notes: This genus was recently described from a specimen found in amber and was included in the current study for completeness, despite the fact that its placement in the Ophiostomatales (Poinar & Vega 2018) is open to debate. The fossil beetle associated with this fungus could have been misidentified and might not be a member of the Platipodinae. Likewise, whether the cavity shown on the beetle reflects a functional mycangium is unclear. The growth form of the fungus growing alongside the beetle is unusual for any beetle-related ophiostomatalean fungus, and mucilage-bound phialidic spores could be those of an asexual morph of an entomopathogenic or another fungus producing phialidica conidiophores.

Raffaelea (Lineage XII)

Raffaelea Arx & Hennebert, Mycopathol. Mycol. Appl. 25: 310. 1965. MycoBank MB 9685, emend. Z.W. de Beer & M. Procter. Fig 13G, H.

Etymology: Named for the Italian mycologist and plant pathologist, Prof. Raffaele Ciferri (1897–1964).

Sexual morph: Ascomatal bases black, subglobose to globose; necks black, straight or curved. Ostiolar hyphae present or absent. Asci evanescent. Ascospores hyaline, 1-celled or rarely 2-celled, cylindrical to oblong, enclosed in hyaline rectangular sheath.

Asexual morph: Colony confluent, mucilaginous. Conidiophores micronematous, macronematous, mononematous, solitary or aggregated in sporodochia, hyaline, simple or branched, monilioid. Conidiogenous cells flask-shaped, proliferating percurrently or sympodially, with denticles, inconspicuous scars or annellations. Conidia hyaline, ellipsoidal, obovoid, globose, pyriform, sometimes T- or Y-shaped, 1-celled, producing secondary cells through budding. Associated with ambrosia beetles.

Type species: R. ambrosiae Arx & Hennebert, Mycopathol. Mycol. Appl. 25: 310. 1965. MycoBank MB 338171.

Other species: Listed in Table 1.

Notes: Raffaelea was originally described to accommodate a number of fungal mutualists of wood-boring ambrosia beetles (Von Arx & Hennebert 1965). In the same paper, Von Arx & Hennebert (1965) validated a similar genus, Ambrosiella, with a mode of conidial development different to that in Raffaelea. However, confusion persisted regarding the generic placement of species in these two genera until DNA sequence data showed that Ambrosiella belonged in the Ceratocystidaceae (Microascales) and Raffaelea in the Ophiostomatales (Batra 1967, Cassar & Blackwell 1996, Massoumi Alamouti et al. 2009, Harrington et al. 2010, De Beer et al. 2013a, 2014). Species of Raffaelea have reduced conidiophore morphology, distinguishing them from other Ophiostomatalean genera (De Beer & Wingfield 2013). All species in the genus are mutualists of ambrosia beetles in the sub-families Scolytinae and Platypodinae (Curculionidae), respectively, in North America, South Africa, Europe and Asia (Harrington et al. 2010, Hulcr & Stelinski 2017).

Prior to the study of Musvuugwa et al. (2015), Raffaelea was considered as an exclusively asexual genus. Musvuugwa et al. (2015) described a Raffaelea species forming sexual structures and emended the generic description of Raffaelea to accommodate these structures. In our analyses, R. vaginata (labelled as Lineage X) did not consistently group within Raffaelea (see below). However, our analyses support the inclusion of R. seticollis in Raffaelea, which was one of the other two sexually reproducing species treated in Raffaelea by Musvuugwa et al. (2015). This species was originally described from ambrosial galleries in a hemlock (Tsuga canadensis) stump in New York State (Davidson 1966) but has not been recorded again. Musvuugwa et al. (2015) also transferred O. deltoideosporum to Raffaelea based on a short LSU sequence of the fungus produced by Hausner et al. (1993b), and morphological similarities to O. seticolle. We did not have access to material of this species and could therefore not include it in our analyses. For the present, we have followed the suggestion of Musvuugwa et al. (2015) and include it in Raffaelea.

Raffaelea incertae sedis (Lineage X)

Raffaelea vaginata

Notes: This species was described from a beetle in the genus Lanurgus collected from a native tree in the Southern Cape of South Africa (Musvuugwa et al. 2015). The fungus formed a sexual morph similar to O. deltoideosporum and O. seticolle that grouped in Raffaelea in the study of De Beer & Wingfield (2013). As stated above, analyses of our data supported the inclusion of R. seticollis (O. seticolle) in Raffaelea. Because R. vaginata did not consistently group within Raffaelea in our analyses, it is not included in our narrower definition of Raffaelea. While it might respresent a novel genus, there is insufficient information to make this decision and it is thus treated as incertae sedis.

Sporothrix (Lineage XIV)

Sporothrix Hektoen & C.F. Perkins, J. Exp. Med. 5: 80. 1900. MycoBank MB 10046, emend. Z.W. de Beer et al., Stud. Mycol. 83: 171. 2016. Figs 14, 15.

Fig. 14.

Genera of the Ophiostomatales redrawn from published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Sporothrix candida complex. C, D. Sporothrix gossypina complex. E, F. Sporothrix inflata complex. G, H. Sporothrix pallida complex. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Fig. 15.

Genera of the Ophiostomatales redrawn published images with sexual morphs (if known) on the left and asexual morphs on the right. A, B. Sporothrix pathogenic clade. C, D. Sporothrix stenoceras complex. (Pale grey shading reflects hyaline to subhyaline colouration, medium-tone grey brown to dark brown and dark grey reflects fuscous black to dark black colouration).

Synonyms: Spumatoria Massee & E.S. Salmon, Ann. Bot. (Oxford) 15: 350. 1901. MycoBank MB 5175. [Type species Spumatoria longicollis Massee & E.S. Salmon, Ann. Bot. (Oxford) 15: 351 (1901)].

Sporotrichopsis Guég., Arch. Parasitol. 15: 104. 1911. MycoBank MB 10048. [Type species S. beurmannii (Matr. & Ramond) Guég.; nom. inval., Art. 38.1].

Dolichoascus Thibaut & Ansel, Compt. Rend. Hebd. Seances Acad. Sci. 270: 2173. 1970. MycoBank MB 1684. (Type species D. schenckii Thibaut & Ansel; nom. inval., Art. 40.1).

Etymology: The name is derived from the Latin for ‘spore hair’ (De Hoog 1974).

Sexual morph: Ascomatal bases black, globose; necks black, becoming paler towards apex, straight or slightly curved. Ostiolar hyphae parallel to divergent. Asci evanescent. Ascospores hyaline, 1-celled, allantoid to reniform in side view, almost triangular-shaped, ovoid to oblong.

Asexual morph: Conidiophores micronematous, semi-macronematous, mononematous, simple. Conidiogenous cells showing sympodial growth, terminal or intercalary, cylindrical, denticulate or not denticulate, hyaline. Conidia hyaline to subhyaline, 1-celled, subglobose to oblong, obovoid, clavate to strongly curved, guttuliform to fusiform with round apices and pointed base that sometimes slightly curved, sometimes slightly curved at base; (pathogenic clade and S. inflata complex) borne directly on hyphae (sessile) brown to dark brown, subglobose to globose, narrowly obovoid, ellipsoid, 1-celled. Secondary conidia borne by yeast-like budding, absent or present.

Type species: Sporothrix schenckii Hektoen & C.F. Perkins, J. Exp. Med. 5: 77. 1900.

Other species: Listed in Table 1.

Notes: Sporothrix was described as part of a medical case study concerning a child who had contracted a fungal infection after injuring his finger with a hammer (Hektoen & Perkins 1900). The fungus isolated from the infection was described as Sporothrix schenckii. The epithet referred to B.R. Schenck who described, but did not name a similar fungus in an earlier study. The genus description of Sporothrix by Hektoen & Perkins (1900) was invalid, and the name was validated years later by Nicot & Mariat (1973). In the late 1960’s, S. schenckii was suggested to represent the asexual morph of O. stenoceras (Nicot & Mariat 1973), and subsequently the asexual morphs of many other Ophiostoma species were also treated in Sporothrix under the dual nomenclature system (De Hoog 1974, De Beer & Wingfield 2013).

The phylogenetic placement of S. schenckii in the Ophiostomatales based on ribosomal DNA sequences by Berbee & Taylor (1992) became the first example where asexual (anamorph) and sexual (teleomorph) genera were connected based on phylogenetic inference. This represented a first important step towards the abandonment of the the dual nomenclature system for the fungi, which culminated in the Amsterdam Declaration (Hawksworth et al. 2011) and subsequent changes to the Code where one fungus could only have one species name.

Following the adoption of the One Fungus-One Name convention, De Beer & Wingfield (2013) listed Sporothrix as a synonym of Ophiostoma, with the type species of Sporothrix (S. schenckii) forming part of the S. schenckii-O. stenoceras complex in a broad concept for Ophiostoma. De Beer & Wingfield (2013) noted that this species complex could represent a genus separate from other species in Ophiostoma s.l. However, the available data at the time were insufficient to formally separate the two genera. In a more comprehensive study including sequences of all Sporothrix spp. and all Ophiostoma spp. with Sporothrix asexual morphs, De Beer et al. (2016a) showed that the S. schenckii-O. stenoceras complex formed a well-supported monophyletic lineage, distinct from other species in Ophiostoma. Because this lineage accommodated the type species of Sporothrix, they reinstated the genus, transferring all Ophiostoma species in this lineage to Sporothrix. De Beer et al. (2016a) also emended the description of Sporothrix to include sexual morphs. Furthermore, they defined several species complexes within Sporothrix. The analyses emerging from the present study support the separation of Sporothrix and Ophiostoma based on strong monophyletic lineages. There are a few exceptions where species formed independent lineages and these are discussed below.

Species complexes:

See De Beer et al. (2016a) and below.

The S. candida complex

The S. candida complex accommodates four species that have been described from hardwoods in South Africa (Kamgan Nkuekam et al. 2012, Musvuugwa et al. 2016) and one species from hardwoods in Argentina (De Errasti et al. 2016). Sporothrix itsvo was previously included in this species complex, but it grouped within the S. gossypina/S. stenoceras complex in our phylogenetic analyses (Fig. 5).

The S. inflata complex

The S. inflata complex accommodates species that have been isolated from oak in Europe, soil in Europe, Canada and Malaysia (De Beer et al. 2016a). It also includes the epitype isolate of Spumatoria longicollis, a coprophilous fungus. In a recent study forming part of The Genera of Fungi project (http://www.mycobank.org), Giraldo et al. (2017) showed an isolate of the monotypic genus Spumatoria (Sp. longicollis) grouping within Sporothrix, based on ITS, LSU and BT sequence data. These authors considered this as the first report of the fungus after it was initially described. The type of this species has been lost (De Beer et al. 2014) and consequently Giraldo et al. (2017) designated an epitype, isolated from cow dung in the Netherlands, which was used in the present study. Although this species has a sporothrix-like asexual morph, its septate ascospores distinguished it from all other ophiostomatalean species. Furthermore, its coprophilous nature and pale coloured ascomata distinguish it from other Sporothrix species (De Beer et al. 2014, Giraldo et al. 2017). De Beer et al. (2013a) excluded Spumatoria from the Ophiostomatales, based on these atypical characteristics and characteristics shared with Kathistes, which is phylogeneticially distant from the Ophiostomatales. Our analyses included LSU and TEF-1α sequence data for the epitype isolate, which placed it within the S. inflata species complex (Figs 5, S1, S3).

The S. stenoceras and S. gossypina complexes

The S. stenoceras and S. gossypina species complexes formed an aggregated lineage in the analyses of our datasets (Figs 5, S1, S3). Interestingly, the lineage within the S. stenoceras complex including species isolated from Protea (De Beer et al. 2016a, Ngubane et al. 2018), and S. gemella (also from Protea), formed a lineage distinct from the other species in these complexes, none of which are associated with Protea.

The S. gossypina complex accommodates species that have diverse ecologies (De Beer et al. 2016a). Some species are associated with conifer-infesting bark beetles (Davidson 1971, Marmolejo & Butin 1990, Zhou et al. 2004a, 2006, Lu et al. 2009, Linnakoski et al. 2010, Six et al. 2011, Taerum et al. 2013), others specifically with pine-infesting bark beetles (Davidson 1971, Zhou et al. 2006, Romón et al. 2014a, b), and a few species that have been isolated from stained oak wood (Aghayeva et al. 2004). Interestingly, there is also a species isolated from cankers on chestnut caused by the aggressive tree pathogen Cryphonectria parasitica (Davidson 1978). Species are widely distributed in the USA, Europe, Asia and South Africa (De Beer et al. 2016a).

Sporothrix stenoceras has been isolated from soils and hardwoods on many continents. This could explain why it grouped with species in the S. gossypina complex.

Sporothrix gemella was previously included in the S. pallida complex, but it grouped within the complex in the analyses of our combined dataset (Fig. 5). This species is found on Protea species in South Africa and is vectored by mites, a niche similar to that of S. splendens (Roets et al. 2013, De Beer et al. 2016a).

Ophiostoma ponderosae grouped in this species complex in the analyses of our combined dataset (Fig. 5). The LSU sequence generated by De Beer et al. (2016a) for the same isolate placed this species in Ophiostoma s.s. Our sequence data for O. ponderosae differed from the sequence generated by De Beer et al. (2016a) in only a small number of bases.

The S. pallida complex

Three species forming part of the S. pallida complex (S. pallida, S. mexicana and S. chilensis) have been reported as rare and opportunistic pathogens of humans, but mostly occurring in soil along with other species in this complex. Two species are associated with Protea species occurring in South Africa. Species in this complex have been found in Japan, Mexico, Chile, England and South Africa (De Beer et al. 2016a).

The pathogenic clade

Most Sporothrix species are relatively harmless to humans, but there are four species (S. schenckii, S. luriei, S. brasiliensis and S. globosa) that are human and animal pathogens. These species are the causal agents of the disease known as sporotrichosis (Teixeira et al. 2014). All four species form pigmented blastoconidia, in addition to the more typical sporothrix-like morphology of the asexual morphs. This is possibly an adaptation to both the soil inhabiting and animal-pathogenic lifestyle of these fungi (De Beer et al. 2016a). Collectively, these pathogens have an almost global distribution. Sporothrix schenckii has been isolated in the Americas, Australia, Southern Africa and Europe, and S. globosa in Central America and parts of Eurasia. However, S. brasiliensis is known only from Brazil, where it is responsible for serious epidemic (Rodrigues et al. 2013, 2014). Sporothrix luriei is known based on only a single isolate from South Africa (Zhang et al. 2013, Teixeira et al. 2014).

Group D

Sporothrix polyporicola grouped alone in Sporothrix in our analyses. (Fig. 5). This is consistent with the phylogenies of Osorio et al. (2016) and De Beer et al. (2016a).

Group E

In a similar manner, S. polyporicola and S. phasma grouped alone in Sporothrix in our analyses (Fig. 5). Both species were closely related in the LSU dataset of De Beer et al. (2016a).

Group F

Group F accommodated S. nigrograna, S. curviconia, S. thermara, S. eucalyptigena and S. bragantina. Sporothrix thermara and S. bragantina grouped together as “Lineage H” and S. curviconia as “Lineage G” in the phylogenies of De Beer et al. (2016a).

Group G

Sporothrix dombeyi formed a single lineage basal to Sporothrix in our concatenated dataset (Fig. 5). Previously S. dombeyi (as O. nothofagi) was transferred to Sporothrix and renamed by De Beer et al. (2016a). This species also grouped basal to Sporothrix in their datasets (along with other species treated here as Lineage XVI, XVIII, XIX).

New combination:

1) Sporothrix longicollis (Massee & E.S. Salmon) M. Procter & Z.W. de Beer, comb. nov. MycoBank MB 841005.

Basionym: Spumatoria longicollis Massee & E.S. Salmon, Ann. Bot. 15: 351. 1901. MycoBank MB 171713.

Description: Massee & Salmon (1901: 350–351, fig. 27), Giraldo et al. (2017: 344–345, fig. 9).

Phylogenetic data: Giraldo et al. (2017).

Notes: Sporothrix longicollis is different from other Sporothrix spp. by having 1-septate ascospores, pale-coloured ascomata and coprophilous biology. The inclusion of this species in Sporothrix was also suggested by Giraldo et al. (2017).

Sporothrix incertae sedis (Lineages XVI & XIX)

Sporothrix fumea (Lineage XVI)

Sporothrix fumea grouped basal to all the above-mentioned lineages in the analyses of our combined dataset (Fig. 5). In the phylogenies of De Beer et al. (2016a), S. fumea formed a lineage with O. valdivianum (Lineage XVIII), but in our phylogenies these species grouped separately.

Sporothrix brunneoviolaceae (Lineage XIX)

In the phylogenies of De Beer & Wingfield (2013), S. brunneoviolaceae formed a distinct lineage in Ophiostoma s.l. with S. fumea and O. fasciatum but these were not available for the present study. However, this species grouped basal to Sporothrix in the phylogenies of De Beer et al. (2016a). In the analyses for the present study, S. brunneoviolaceae consistently grouped separately from Sporothrix.

CONCLUSIONS

The results of this study have provided support for the delineation of most of the genera that have been recognised during the course of the last decade in the Ophiostomatales. This study also revealed robust novel lineages described here in four new genera. In addition to phylogenetic support, most of these genera include coherent groups of species that are characterised by similar morphology, ecology and/or geographical origin. The study has also resolved remaining One Fungus-One Name issues that were not dealt with in the revision of De Beer & Wingfield (2013). This is especially by providing new combinations for several species in Grosmannia and Leptographium.

This study is the most extensive and most comprehensive ever undertaken on the taxonomy of the Ophiostomatales. The concatenated dataset included 264 taxa representing all major lineages in the Order. It was not possible to resolve the taxonomic status of some smaller lineages. These lineages may represent new genera or reside in existing genera that will emerge as new species are discovered. We have consequently chosen to retain species in these lineages in their current genera until such data can provide clarity on their taxonomic placement.

The phylogenies generated in this study will serve as a framework for future taxonomic studies on the Ophiostomatales. In the short term, they will facilitate the appropriate generic placement of novel species. But they will also provide the required lists of the appropriate species to include in smaller phylogenies of single genera and/or species complexes that will make it possible to confirm whether studied isolates represent novel taxa.

Supplementary Material: https://studiesinmycology.org/

Fig. S1.

Phylogenetic tree derived from maximum likelihood analysis of the LSU gene region. The dataset consisted of 233 isolates and 859 characters before and after Gblocks treatment. Bootstrap values above 60 % are shown. Purple blocks indicate existing genera, yellow blocks new genera described in this study, green blocks genera that we redefine here, and blue blocks indicate genera that we here re-instate and re-define.

Fig. S2.

Phylogenetic tree derived from maximum likelihood analysis of the ITS gene region. The dataset consisted of 235 isolates and 169 characters after Gblocks treatment (1 171 characters prior to Gblocks, including gaps). ML values above 60 % are shown. Purple blocks indicate existing genera, yellow blocks new genera described in this study, green blocks genera that we redefine here, and blue blocks indicate genera that we here re-instate and re-define.

Fig. S3.

Phylogenetic tree derived from maximum likelihood analysis of the TEF1-α gene region. The dataset consisted of 207 isolates and 380 characters after Gblocks treatment (655 characters prior to Gblocks, including gaps). ML values above 60 % are shown. Purple blocks indicate existing genera, yellow blocks new genera described in this study, green blocks genera that we redefine here, and blue blocks indicate genera that we here re-instate and re-define.

Fig. S4.

Phylogenetic tree derived from maximum likelihood analysis of the RPBII gene region. The dataset consisted of 171 isolates and 952 characters after Gblocks treatment (1 108 characters prior to Gblocks, including gaps). ML values above 60 % are shown. Purple blocks indicate existing genera, yellow blocks new genera described in this study, green blocks genera that we redefine here, and blue blocks indicate genera that we here re-instate and re-define.

Table S1.

Taxa included in the phylogenomic analyses and their genome sequence statistics