Ophiostoma babimostense and Sporothrix europaea (Ascomycota, Ophiostomatales), two new ophiostomatalean species, associated with ambrosia and bark beetles in Norway and Poland

121 Ophiostoma babimostense and Sporothrix europaea (Ascomycota, Ophiostomatales), two new ophiostomatalean species, associated with ambrosia and bark beetles in Norway and Poland Robert Jankowiak1, Halvor Solheim2, Piotr Bilański1, Filip Kawa1 1 Department of Forest Ecosystems Protection, University of Agriculture in Krakow, Al. 29 Listopada 46, 31-425 Krakow, Poland 2 Norwegian Institute of Bioeconomy Research, P.O. Box 115, 1431 Ås, Norway Corresponding author: Robert Jankowiak (rober[email protected]) Copyright: © Robert Jankowiak et al. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International – CC BY 4.0). Research Article Abstract The order Ophiostomatales includes many species important for forestry, causing plant diseases. They are common associates of barkand wood-dwelling beetles. Two new ophiostomatalean fungi viz. Ophiostoma babimostense sp. nov. and Sporothrix europaea sp. nov. are proposed, based on morphological characters and multigene phylogenies. Ophiostoma babimostense belongs to the Ophiostoma ulmi species complex and was isolated from fallen shoots of Scots pine pruned by Tomicus species in Poland. The fungus is characterised by the production of a typical pesotum-like and sporothrix-like asexual morphs. Sporothrix europaea belongs to the Sporothrix gossypina complex and was isolated from hardwood-infested by ambrosia and bark beetles in Poland and Norway. It is characterised by the occurrence of both a sexual and asexual morphs, with long necked ascomata bearing ostiolar hyphae and a sporothrix-like asexual morph. Key words: Beetle-associated fungi, hardwood, phylogenetics, Pinus sylvestris, taxonomy, two new taxa Introduction Ophiostomatalean fungi (Ophiostomatales, Ascomycota) are characterised by the formation of flask-shaped perithecia and various asexual morphs, including mononematous or synnematous forms (Wingfield et al. 1993; De Beer and Wingfield 2013). These fungi include treeor wood-infecting species causing a dark bluish discoloration in the sapwood or serious tree diseases and some of them are also the causal agents of human diseases. Ophiostomatalean species are well known to be closely associated with barkand wood-dwelling beetles and mites (De Beer et al. 2022). In Europe, surveys for ophiostomatalean fungi from different habitats are constantly improving. Combined with multi-locus molecular phylogenetic analysis has led to the identification of many species of ophiostomatalean fungi, including dozens of novel species (e.g. Linnakoski et al. (2016); Jankowiak et al. (2017, 2019b)). In particular, extensive surveys have been conducted on the diversity and taxonomy of ophiostomatalean and microascalean fungi in CenAcademic editor: Thorsten Lumbsch Received: 11 April 2025 Accepted: 18 September 2025 Published: 13 October 2025 Citation: Jankowiak R, Solheim H, Bilański P, Kawa F (2025) Ophiostoma babimostense and Sporothrix europaea (Ascomycota, Ophiostomatales), two new ophiostomatalean species, associated with ambrosia and bark beetles in Norway and Poland. MycoKeys 123: 121–145. https://doi. org/10.3897/mycokeys.123.155588 MycoKeys 123: 121–145 (2025) DOI: 10.3897/mycokeys.123.155588 122 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland tral and Northern Europe. For example, surveys of insect-associated mycobiomes in Norway and Poland, yielded descriptions of almost 30 new species of Ceratocystiopsis (Jankowiak et al. 2022), Graphilbum (Jankowiak et al. 2020), Graphium (Jankowiak et al. 2023a), Ophiostoma (Aas et al. 2018; Jankowiak et al. 2019a) and Sporothrix (Ostafińska et al. 2021). More recent discoveries generated descriptions of a new genus Hausneria (Crous et al. 2024) from a gallery of Dryocoetes alni in Norway and the new species, Ophiostoma juglandis from Dryocoetes himalayensis on Juglans regia in Czechia (Májek et al. 2025). In Poland, the screening of ophiostomatalean fungi was conducted from tree wounds (Jankowiak et al. 2019c), in forest soils (Bilański et al. 2023) and in bird’s nests (Błońska et al. 2021), yielding two new soil-inhabiting Sporothrix species (Bilański et al. 2023), one new species of Hawksworthiomyces from nests of Ciconia ciconia (Crous et al. 2023) and three new species of Leptographium from tree wounds (Jankowiak et al. 2018a). These surveys revealed a plethora of new beetle-fungus associations and led to the discovery and description of many new species, showing there is still a lot of unknown taxonomic and ecological diversity to be uncovered for the ophiostomatalean fungi in Europe. Ophiostoma is one of the largest genera within the Ophiostomatales and includes treeor wood-infecting fungi. Currently, more than 170 species are recognised worldwide (De Beer et al. 2022; Bhunjun et al. 2024; Feau et al. 2024; Májek et al. 2025; Wang et al. 2024). Members of Ophiostoma are characterised by pigmented ascomata with slender necks and allantoid, reniform, cylindrical to ossiform in side view ascospores. In addition, Hyalorhinocladiella-, Leptographium-, Pesotumor Sporothrix-like asexual morphs are produced in culture and nature (De Beer et al. 2022). Ophiostoma is mostly structured into six phylogenetically well-supported lineages or so-called species complex. They are the O. clavatum–, O. ips–, O. minus–, O. piceae–, O. pluriannulatum– and O. ulmi lineages and species complexes. In addition, some species do not form part of these lineages, such as O. piliferum and O. tetropii (De Beer et al. 2022; Wang et al. 2024). Ophiostoma can be found in a wide range of habitats, but most species are commonly associated with forest trees on which they form more or less stable associations together with barkand wood-dwelling beetles, mites (Six 2012) and, in a few cases, nematodes (Bhunjun et al. 2024). Some members of Ophiostoma, such as O. piliferum and O. minus, cause the economically important blue-stain in freshly exposed sapwood of softwood species (e.g. Seifert (1993); Uzunović and Byrne (2013); Jankowiak et al. (2021)) or sawn timber (Jankowiak et al. 2018b). In addition, O. novo-ulmi is an example of a highly virulent pathogen that has been responsible for Dutch elm disease in Europe, western Asia and North America (Brasier 1991). Ophiostoma has a worldwide distribution, but it is especially abundant in the north temperate to boreal conifer ecosystems, in Asia, Europe and North America. They have been observed from Abies, Larix, Picea, Pinus, Pseudotsuga and Tsuga. Ophiostoma species also grow from hardwood, such as Quercus or Fagus (Jankowiak et al. 2019b, c). Some species, such as Ophiostoma ips, O. piceae, O. piliferum and O. quercus, are globally widespread, possibly due to human activity and the movement of wood products around the globe (Taerum et al. 2018; Bhunjun et al. 2024). Sporothrix is also a large genus in the Ophiostomatales, with currently 70 recognised species arranged in five phylogenetically well-supported lineages, forming species complex, including the S. candida–, S. inflata–, S. stenoceras–, 123 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland S. gossypina– and S. pallida species complexes (De Beer et al. 2016, 2022; Wang et al. 2019; Bilański et al. 2023). Other species form part of several unresolved smaller phylogenetic lineages (Groups D-G) or two lineages Sporothrix insertae sedis (XVI & XIX) defined by De Beer et al. (2022). The S. gossypina complex is considered as the largest at present, with 18 accepted species. Members of this complex are characterised by the presence of globose ascomatal base with black necks and allantoid to reniform in side view ascospores. The asexual morphs include simple, micronematous to mononematous conidiophores, with denticulate or not denticulate conidiogenous cells showing sympodial growth. Conidia hyaline to that vary in size and shape, mostly subglobose to oblong, obovoid, clavate to strongly curved, guttuliform to fusiform (De Beer et al. 2022). Similar to Ophiostoma, members of Sporothrix are widely distributed across various climatic zones of the world, colonising diverse environments (De Beer and Wingfield 2013; De Beer et al. 2016). The greatest numbers of species are found on the bark and wood of different forest trees and in the infructescences of Protea spp. (e.g. Roets et al. (2013); De Errasti et al. (2016); Bilański et al. (2023)). Other species have been described from soil, ambrosia and bark beetles, mites and from the fruiting bodies of basidiomycetes (e.g. Constantinescu and Ryman (1989); Marmolejo and Butin (1990); De Meyer et al. (2008)). Several species are also well-known as human and animal pathogens (e.g. Lòpez-Romero et al. (2011); Zhang et al. (2015)). During a survey of ophiostomatalean fungi on hardwoods in Poland and Norway (Aas et al. 2018; Jankowiak et al. 2019b), isolates of an undescribed Sporothrix species with a sexual state resembling species in the S. gossypina species complex were isolated from different ambrosia and bark beetle species. In addition, isolates of an unknown fungal species with morphological attributes (synnemata) matching those described for species of the Ophiostoma were isolated from Scots pine (Pinus sylvestris L.) shoots infested by an unknown species of Tomicus. The purpose of this study is to characterise these fungi using the morphology, phylogenetic analyses and to formally describe them as novel species of Sporothrix and Ophiostoma. Materials and methods Isolates and herbarium specimens A collection of ten Polish and three Norwegian isolates were used in this study. Isolates of Ophiostoma sp. were collected in pure Scots pine stands in western Poland (Babimost: 15°50′33″N, 52°09′11″E) in October 2023. Isolations were made from fallen shoots of Scots pine pruned by Tomicus spp. as described by Jankowiak and Kolařík (2011). Isolates of Sporothrix sp. were collected during surveys of hardwood-infesting bark and ambrosia beetles in Poland and Norway (Aas et al. 2018; Jankowiak et al. 2019b). The cultures are maintained in the culture collection of the Department of Forest Ecosystems Protection, University of Agriculture in Krakow, Poland and in the culture collection of Norwegian Institute of Bioeconomy. The ex-type isolates and representative isolates of the new species described were deposited in the culture collection (CBS) of the Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands and in the culture collection (CMW) of the Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, 124 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland South Africa. Dried cultures were deposited as holotype specimens in the National Biodiversity Collection – Herbarium KRAM, the W. Szafer Institute of Botany Polish Academy of Science, Kraków, Poland. Two reference strains were also obtained from collections of CBS. These included a living culture of Sporothrix fusiformis (CBS 112912) and Sporothrix lunata (CBS 119444) (Table 1). PCR, sequencing and phylogenetic analyses DNA extraction, PCR and sequencing were performed as described by Jankowiak et al. (2019a). For DNA sequencing and phylogenetic analyses, five loci were amplified as follows: the internal transcribed spacer regions ITS1-5.8SITS2 (ITS), the partial 28S ribosomal large subunit (LSU), the partial beta-tubulin (TUB2), the partial calmodulin (CAL) and the partial translation elongation factor 1-alpha (TEF1). The primers used for polymerase chain reaction (PCR) and sequencing of the various gene regions were as follows: ITS1-F (Gardes and Bruns 1993) and ITS4 (White et al. 1990) for ITS, LR5 and LR0R (Vilgalys and Hester 1990) for LSU, Bt2a and Bt2b (Glass and Donaldson 1995) for TUB2, EF2F (Marincowitz et al. 2015) and EF2R (Jacobs et al. 2004) or F-728F (Carbone and Kohn 1999) and EF2 (O’Donnell et al. 1998) for TEF1 and CL1 and CL2a (O’Donnell et al. 2000) or CL3F and CL3R (De Beer et al. 2016) for CAL. BLAST searches using the BLASTn algorithm were performed to retrieve similar sequences from GenBank (http://www.ncbi.nlm.nih.gov) and accession numbers for these sequences are presented in the corresponding phylogenetic trees (Figs 1–4). Datasets were curated using Molecular Evolutionary Genetic Analysis (MEGA) 6.06 (Tamura et al. 2013). The ITS datasets included all available sequences for reference species of Ophiostoma and Sporothrix that could be retrieved from GenBank to resolve the placement of the isolates within the mentioned genera. Sequence alignments were performed using the online version of MAFFT 7 (Katoh and Standley 2013) using the E-INS-i strategy with a 200PAM/κ = 2 scoring matrix, a gap opening penalty of 1.53 and an offset value of 0.00. The alignments were checked manually with BioEdit 2.7.5 (Hall 1999). The resulting alignments and trees were deposited in TreeBASE (http://purl.org/phylo/treebase/phylows/study/TB2:S32067). All phylogenetic analyses were performed independently for each gene regions (ITS, TUB2, TEF1 for Ophiostoma and ITS, CAL, TUB2 for Sporothrix). Sequences of the LSU (for Ophiostoma, Sporothrix), CAL (for Ophiostoma) and TEF1 (for Sporothrix) genes were generated for future reference. Gene regions showing no conflicts in the grouping of isolates at the terminal clades (i.e. TUB2 and TEF1 for Ophiostoma and TUB2 and CAL for Sporothrix), were combined and analysed as concatenated datasets. Phylogenetic trees were inferred for each of the datasets using three different methods: Maximum Likelihood (ML), Maximum Parsimony (MP) and Bayesian Inference (BI). For ML and BI analyses, the best-fit substitution models for each dataset were determined using the corrected Akaike Information Criterion (AICc) in jModelTest 2.1.10 (Guindon and Gascuel 2003; Darriba et al. 2012). ML analyses were carried out using PhyML 3.0 (Guindon et al. 2010), utilising the Montpelier online server (http://www.atgc-montpellier.fr/phyml/). The ML analysis included bootstrap analysis (1000 bootstrap pseudoreplicates) in order to assess node support values. 125 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland 0.3 O. piceae CMW25034 KU184441 O. denticiliatum CMW29493 FJ804490 726J PV350321 O. brunneum CMW1027 KU184423 O. signatum CBS144267 MH055643 O. hylesinum CBS144296 MH055636 O. novo-ulmi subsp. novo-ulmi CBS298.87 AF198235 O. shanziensis CMW48329 MT637221 O. undulatum CMW19396 GU797218 O. breviusculum YCC-519 AB200421 O. catonianumCBS263.35 AF198243 O. minus CMW43873 OM501497 O. canumCMW29495 KU184424 O. peregrinum AE426 MG345116 O. haidaense MB13A OR437222 Ophiostoma tasmaniense CMW29088 GU797211 836J PV350325 O. minus OM1 AY542494 O. album CXY1622 KY094073 CBS152112 PV350324 O. perfectum CMW17153 OM501506 O. novo-ulmi subsp. novo-ulmi CBS144287 MH055667 O. gmelinii CMW40463 MW581495 O. allantosporum CBS185.86 AY934506 O. flexuosum CMW907 KU184427 O. himal-ulmi CBS374.67 AF198233 O. tsotsi CMW15239 FJ441287 O. nitidumCMW38905 KU184436 O. patagonicum CIEFAP465 KT362247 O. distortum CBS429.82 KU184426 O. taizhouense CFCC55740 OK104005 O. quercus CMW2465 MH248734 O. olgensis CXY1410 KU551303 O. huangnanens CFCC55624 MZ328850 CBS152111 PV350322 O. villosum CBS144272 MH055648 O. cupulatum CMW37441 OM501479 O. pseudominus CMW43878 OM501508 O. nikkoense CMW17194 KU184435 746J PV350323 O. rufum CBS144871 MH837040 O. xinganense CFCC52679 MK748186 O. pseudotsugaeD483 AY542501 O. jingoulingense CFCC57826 PP656989 O. pseudokarelicum CBS144275 MH055652 O. setosum CMW27834 KU184452 O. xiaojinense CFCC57836 PP656993 O. kryptum DAOM229701 AY304436 O. taphrorychi CBS144891 MH837052 O. bacillisporum MUCL45378 AY573258 O. karelicum CMW23094 EU443759 O. floccosum CBS799.73 AF198231 O. micansCMW38909 KU184433 O. introcitrinum CMW1601 OM501485 O. tibetense CFCC57832 PP656992 O. tetropii CBS428.94 AY934524 O. multisynnematum CFCC52677 MK748196 O. novo-ulmi subsp. americanaC510 AF198236 O. torulosum CMW10574 OM501518 O. nyangense CFCC57830 PP656990 O. australiae CMW6606 EF408603 O. ulmi CBS102.63 AF198232 O. wuyingense CMW44474 MH144061 O. qinghaiense CMW38906 KU184447 O. sugadairense YCC589 LC090226 O. pityokteinis CBS144879 MH837046 O. typographiCMW44483 MH144059 O. rachisporum CMW23274 KU184450 O. minus 1412 MK748202 O. borealis CMW18966 EF408593 O. genhense CFCC52675 MK748199 */100 99/94 */100 100/100 */100 */100 */100 98/* */100 */100 */100 */100 */75 */100 100/100 */100 97/92 */100 */* */100 96/97 100/100 100/100 100/93 */100 O. babimostense sp. nov. O. ulmi species complex O. minus species complex Group B Group K O. piceae species complex Figure 1. Phylogram from Maximum Likelihood (ML) analysis of ITS data for Ophiostoma spp. Polish isolates used in this study are in bold. Bootstrap values (if ≥ 75%) for ML and Maximum Parsimony (MP) analyses are presented at the nodes as follows: ML/MP. Bold branches indicate posterior probabilities values ≥ 0.95 obtained from Bayesian Inference (BI) analysis. * Bootstrap values < 75%. The tree is drawn to scale (see bar) with branch lengths measured in the number of substitutions per site. The species complexes were designated, based on the classification of De Beer et al. (2022) and Wang et al. (2024). Heinzbutinia solheimii and H. grandicarpa represent the outgroup. 126 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Figure 1. 0.3 O. adjunctiCMW1025 OM501462 O. maixiuense CFCC55627 MZ328852 O. macroclavatum CMW23115 HM031499 Heinzbutinia grandicarpa CBS250.88 KX590820 O. aggregatum CXY1876 MH555894 O. clavatum CMW37983 KU094685 O. poligraphiCMW38898 KU184443 O. montium CMW13221 AY546711 O. hongxingense CFCC52695 MK748194 O. fuscum CMW23195 OM501483 O. qinghense CFCC57854 PP657010 O. longiconidiatum CMW17574 EF408558 O. daofuense CFCC57746 PP656972 O. stebbingiCFCC57770 PP656979 O. tingens CBS366.53 MH857246 O. subannulatum CBS188.86 AY934522 O. palustre CMW44427 KU865595 O. pseudocatenulatum CMW43103 KU094686 O. piliferum JGI MT633063 O. multiannulatum MUCL19062 AY934512 O. brunneolum CMW23143 KU094684 O. ips CMW19371 OM501486 O. gilletteae CMW30681 MT637227 O. jiamusiensis CMW40512 MH144064 O. sanumCFCC55632 MZ328857 O. shennongense CFCC53921 MW459989 O. sparsiannulatum CMW17231 FJ906817 O. kunlunense CMW41927 MH121648 O. juglandis CCF6633 OR946482 O. manchongiCMW41954 MH121662 O. pluriannulatum CMW22803 DQ539508 O. araucariae CBS114.68 KU184418 O. schmutzenhoferi CFCC57762 PP656977 O. miyingense CFCC57753 PP656974 O. pacis CFCC57849 PP657005 O. shangrilae CBS136520 KU184453 O. ainoae CMW1903 HM031495 O. songshuiCMW44473 MH144065 O. californicum CBS796.73 KU756602 O. pseudobrevipilosi CFCC57760 PP656976 O. carpenteri CMW44611 OM501475 O. subelongatiCFCC52693 MK748200 O. armandii CFCC57737 PP656970 O. sexdentatiCFCC57812 PP656986 O. hongshiense CFCC57749 PP656973 O. bicolor CBS492.7 DQ268604 O. ponderosae CMW37953 OM501554 O. piliferi CXY4048 OM397409 O. brunneociliatum CMW39827 KU094683 O. altaiense CFCC57840 PP656996 O. yaluense CXY4059 OM397414 O. tonghuaense CXY4055 OM397412 O. novae-zelandiae CIEFAP423 KT362249 O. peniculi CFCC52687 MK748198 O. conicola CBS127.89 OM501478 O. japonicum YCC099 GU134169 O. brevipilosi CMW41662 MG205660 O. macrosporum CMW14176 OM501495 Heinzbutinia solheimii CBS144881 MH283134 O. triangulosporum DSMZ4934 AY934525 O. pseudobicolor CFCC52683 MK748188 O. tapionis CMW23269 HM031494 O. pulvinisporum CMW9022 AY546714 O. shigatseense CFCC57815 PP656987 O. yadongense CFCC57775 PP656981 86/86 77/* */* */* */* */* */76 99/92 93/98 77/* */86 100/100 */* 100/94 100/100 */* */100 94/98 */100 100/100 */100 87/99 96/98 */100 */98 */100 79/* 100/100 */* */* 97/97 100/100 */* */* 100/95 ×10 ×10 // // 100/100 O. pluriannulatum species complex Group H O. clavatum species complex Group C Group I O. ips species complex Group J Figure 1. Continued. MP analyses were performed using PAUP* 4.0b10 (Swofford 2003). Gaps were treated as the fifth state. Bootstrap analysis (1000 bootstrap replicates) was conducted to determine the levels of confidence for the nodes within the inferred tree topologies. Tree bisection and reconnection (TBR) was selected as the branch swapping option. The tree length (TL), consistency index (CI), retention index (RI), homoplasy index (HI) and rescaled consistency index (RC) were recorded for each analysed dataset after the trees were generated. 127 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Figure 2. Phylogram from Maximum Likelihood (ML) analysis of the combined datasets of TUB2+TEF1 for the Ophiostoma ulmi species complex. Polish isolates used in this study are in bold. Bootstrap values (if ≥ 75%) for ML and Maximum Parsimony (MP) analyses are presented at the nodes as follows: ML/MP. Bold branches indicate posterior probabilities values ≥ 0.95 obtained from Bayesian Inference (BI) analysis. * Bootstrap values < 75%. The tree is drawn to scale (see bar) with branch lengths measured in the number of substitutions per site. Ophiostoma tapionis represents the outgroup. 0.2 O. novo-ulmi CBS144287 836J O. quercus CMW2467 O. quercus CBS144291 O. catonianumCBS263.35 O. australiae CMW6589 O. villosum CBS144274 O. novo-ulmi CMW1463 Ophiostoma tasmaniense CMW29115 O. tasmaniense CMW3196 746J O. novo-ulmi CBS144289 O. patagonicum CMW38089 O. signatum CBS144268 O. undulatum CMW19397 O. himal-ulmi CMW22729 O. pseudokarelicum CBS144278 O. signatum CBS144269 O. tsotsi CBS123599 O. bacillisporum CBS771.71 O. australiae CMW6606 O. araucariae CBS114.68 O. tsotsi CMW3117 O. undulatum CBS127183 O. bacillisporum CMW2579 O. karelicum KFL97116RJSR O. ulmi W9 O. tapionis CMW23265 O. tasmaniense CBS127212 O. ulmi 785401 CBS152111 O. boreale CMW18966 O. hylesinumCBS144262 O. villosum CBS144272 O. novo-ulmi CMW10573 O. hylesinum CBS144296 O. pseudokarelicum CBS144281 CBS152112 O. tapionis 133SSW 726J O. denticiliatum KFL1075NLP16RJ O. karelicum CBS144283 100/100 100/100 100/98 100/100 100/100 91/96 */* 100/100 */* 80/98 100/100 100/100 100/100 99/97 100/100 94/83 100/100 100/100 98/96 95/78 100/100 100/100 100/100 100/100 100/100 100/87 100/96 100/100 100/100 96/88 98/98 100/100 // ×10 // O. babimostense sp. nov. ×10 BI analyses using Markov Chain Monte Carlo (MCMC) methods were carried out using MrBayes 3.1.2 (Ronquist and Huelsenbeck 2003). Four MCMC chains were run for 10 million generations, applying the best-fit model for each dataset. Trees were sampled every 100 generations, resulting in 100,000 trees. Tracer 1.4.1 (Rambaut and Drummond 2007) was utilised to determine the burn-in value for each data-set. The remaining trees were utilised to generate a 50% majority rule consensus tree, which allowed for calculating posterior probability values for the nodes. Morphology and growth rate Morphological characters were examined for selected isolates as well as for the herbarium specimens selected as types. Cultures were grown on 2% malt extract agar (MEA) made up of 20 g malt extract, 20 g bacteriological lab-agar (Biomaxima S.A., Lublin, Poland) in 1 litre deionised water. Fungal cultures were 128 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland 0.4 CBS151676 PV350327 S. fraxini CBS147936F MH283150 S. resoviensis CBS147927F MH740962 S. humicola CMW7618 AF484472 S. oleae CMW40361 MN298850 S. fusiformis KFL57016RJAD MH283155 S. fusiformis CBS149833 MH283159 S. fusiformis CBS149834 MH283158 S. silvicola CBS149241 OP594842 S. itsvo CMW40370 KX590840 S. guttiliformis CBS437.76 KX590839 S. protearum CMW1107 DQ316201 S. stenoceras CBS237.32 AF484464 S. prolifera CBS251.88 KX590829 S. abietina CBS125.89 AF484453 S. fusiformis CBS112912 AY280481 S. pallida CBS131.56 EF127880 S. euskadiensis CMW27318 DQ674369 S. chilensis CBS139891 KP711811 S. eucastanea CBS424.77 KX590814 S. macroconidia CFCC52628 MH555898 S. rossii CBS116.78 KX590815 Sporothrix lunata CBS112927 AY280485 S. schenckii CBS359.36 AY280495 S. cantabriensis CMW39766 KF951554 CBS149831 KY568169 S. uta CMW40316 OM501565 S. africanaCMW823 DQ316197 S. roztoczensis CBS147973 OP594847 S. palmiculminata CMW20677 DQ316191 S. fusiformis CBS149835 MH283154 C. neglecta CBS100596 MH862711 S. aurorae CBS118837 DQ396796 S. gossypina ATCC18999 KX590819 S. nsini CMW28602 EU660458 S. aemulophila CMW40381 OM501527 CBS151675 PV350326 S. dentifundaCBS115790 AY495434 S. cracoviensis CBS147942 MW768964 S. pseudoabietina CFCC52626 MH555896 S. stylites CMW14543 EF127883 S. mexicanaCBS120341 KX590841 S. protea-sedis CMW28601 EU660449 S. narcissi CBS138.50 AY194510 S. gemella CMW23057 DQ821560 S. cabralii CMW38098 KT362256 S. villosa SNM188 MW989428 S. dimorphospora CBS125442 KP017082 S. zambiensis CBS124912 EU660453 S. globosa CBS120340 KP017086 2016-1712/2/1 PV350328 S. luriei CBS937.72 AB128012 KFL77916RJSR MH283143 S. variecibata CMW23051 DQ821568 S. rostrocoronatum CBS434.77 AY194509 S. splendens CMW23050 OM501561 S. inflata CBS239.68 AY495426 Ceratocystiopsis neglecta CBS147960 OL309945 S. brasiliensis CBS120339 KX590832 CBS149832 MH283145 S. undulataCBS147929 MH740976 KFL39616RJSI MH283142 S. cryptarchumCBS147934 MW768966 S. polyporicola CBS669.88 KX590827 S. cavum CBS147943 MF782813 CBS149830 MH283141 S. tumidusCBS147970 OP594850 */* 100/100 */* 97/96 99/99 86 /80 99/99 75/* 90/* */* */* */* 92/98 97/98 99/99 100/100 87/* */* */99 */96 */* 90/99 84/* 93/83 */* 100/100 */96 76/97 75/* 75/* */* */80 88/93 100/100 88/100 98/82 */* */* 88/76 100/81 89/96 89/95 ×10 // ×10 // Sporothrix europaea sp. nov. S. stenoceras species complex Pathogenic clade S. inflata species complex S. candida species complex S. pallida species complex S. gossypina species complex Group O Figure 3. Phylogram from Maximum Likelihood (ML) analysis of ITS data for Sporothrix spp. Norwegian and Polish isolates used in this study are in bold. Bootstrap values (if ≥ 75%) for ML and Maximum Parsimony (MP) analyses are presented at the nodes as follows: ML/MP. Bold branches indicate posterior probabilities values ≥ 0.95 obtained from Bayesian Inference (BI) analysis. * Bootstrap values < 75%. The tree is drawn to scale (see bar) with branch lengths measured in the number of substitutions per site. The species complexes were designated, based on the classification of De Beer et al. (2016). Ceratocystiopsis neglecta represents the outgroup. 129 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Figure 4. Phylogram from Maximum Likelihood (ML) analysis of the combined datasets of TUB2+CAL for the Sporothrix stenoceras and S. gossypina complexes. Norwegian and Polish isolates used in this study are in bold. Bootstrap values (if ≥ 75%) for ML and Maximum Parsimony (MP) analysis are presented at the nodes as follows: ML/MP. Bold branches indicate posterior probabilities values ≥ 0.95 obtained from Bayesian Inference (BI) analysis. * Bootstrap values < 75%. The tree is drawn to scale (see bar) with branch lengths measured in the number of substitutions per site. Sporothrix brunneoviolacea represents the outgroup. Figure 4. 0.05 S. brunneoviolacea CBS124561 S. africana8.3 KFL39616RJSI CBS149830 S. zambensis CBS124912 CBS151675 S. rossii CBS116.78 S. cracoviensis CBS147941 S. stenoceras CBS237.32 S. variecibata CBS121961 S. gossypina ATCC18999 S. fusiformis KFL43916RJSR S. splendens CBS116379 S. stenoceras KFL17NKAS17 S. prolifera CBS251.88 CBS149832 Sporothrix lunata CBS112927 S. fusiformis CBS149835 CBS151676 S. prolifera KFL218N16TARAO S. eucastaneae CBS424.77 S. eucastaneae KFL1141N16DBRJ S. cracoviensis CBS147942 S. fusiformis KFL57016RJAD S. cantabriensis CMW39767 S. aurorae KFL8NBK17KAS S. villosa SNM188 S. resoviensis CBS147927 S. pseudoabietina CFCC52626 2016-1712/2/1 KFL77916RJSR S. brunneoviolacea KFL41PFDb S. fusiformis CBS112912 S. pseudoabietina CBS147969 S. protearum CBS116654 S. fraxini CBS147938 S. euskadiensis CBS122138 S. cantabriensis CMW39766 S. narcissiCBS138.5 CBS149831 S. fusiformis CBS149834 S. fusiformis CBS112925 S. aurorae CBS118837 S. fusiformis CBS149833 S. fraxini CBS147936 S. abietina CBS125.89 88/88 100/100 */99 88/85 */100 99/100 100/100 100/100 */* */77 100/100 */99 99/95 99/99 99/92 */* 83/89 */* 100/100 100/100 100/100 79/88 96/100 100/100 */99 100/100 ×10 // // */* Sporothrix europaea sp. nov. ×10 derived from single conidia spores. The cultures were incubated at 25 °C and monitored regularly for the appearance of sporulating structures. Attempts were made to induce the formation of sporulating structures, by placing autoclaved twigs of host trees at the centres of agar plates containing 2% MEA. To promote the production of ascomata, all isolates of each species were crossed in all possible combinations, following the technique described by Grobbelaar et al. (2009). These cultures were incubated 25 °C in darkness and monitored regularly for the appearance of sporulating structures. Morphological features were examined by mounting fungal tissue in 80% lactic acid on glass slides and fruiting structures were observed using a Nikon Eclipse 50i microscope (Nikon, Tokyo, Japan) with an Invenio 5S digital camera 136 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland strongly thickened, 10–19 in number, (14–)29–42.5(–48.5) μm long, 0.5–1.5 μm at the apex and 1–2.5 μm at the base. Asci evanescent. Ascospores one-celled, hyaline, allantoid in side view (3–)3.5–4(–5) x (0.5–)1–1.5(–2) μm, elliptical in front view (2.5–)3–4(–5) × (1–)1–1.5(–1.5) μm, sometimes with residual sheath up to 2 μm thick, accumulated in white-colour mass at the tip of the neck. Asexual morph Sporothrix-like: conidiophores hyaline, micronematous, simple or branched and bearing several conidiogenous cells, borne on upright undifferentiated hyphae. Conidiogenous cells cylindrical, terminal or intercalary, straight or curved, tapering towards the apex, (3.5–)15–36(–50.5) μm long, (0.5–)1–2(–2) μm wide at the base, the apical part swollen, (1–)2–3(–3) μm long, (1.5–)2–4(–5.5) μm wide, with multiple conidiogenous loci as denticles, born by sympodial proliferation. Conidia of two types: 1) abundant in cultures, hyaline, unicellular, smooth, variable in shape and size, guttuliform to fusiform, curved, often asymmetric, pointed at the base, (3–)3.5–5(–7)× (0.5–)1–1.5(–2.5) μm, formed directly on denticles; 2) sparse in cultures, directly on the side of submerged hyphae in malt agar, subhyaline to lightly pigmented, unicellular, smooth, subglobose to broadly obovate, (2.5–)3–4(–5.5)× (2–)2–3(–4) μm diam., formed singly. Culture characteristics. Colonies with optimal growth at 25 °C on 2% MEA reaching an average of 47 mm (± 0.07 mm) after 14 days, with radial growth rate 1.68 (± 0.24) mm/d, growth somewhat slower at 30 °C (40 mm diameter); white (3A1), flat, floccose, growing in a circular pattern with entire margins, reverse yellowish-white (3A2). Hyphae greenish-grey (1B7) in colour, smooth, with granules, submerged in the medium and aerial mycelium abundant, (0.5–)1– 1.5(–3) µm wide. Associated insects. Anisandrus dispar, Ips cembrae, Scolytus intricatus, Scolytus rugulosus, Xyleborinus saxesenii, Xyleborus monographus. Host trees. Fagus sylvatica, Larix decidua, Prunus domestica, Quercus robur Distribution. Norway, Poland Additional specimen examined. Poland • Prószków, from Xyleborus monographus infesting Quercus robur, May 2013, coll. P. Wieczorek, (culture CBS 149830=CMW 60554); Norway • Ås, Anisandrus dispar on Quercus robur, June 2016, coll. T. Aas, (culture CBS 151675). Notes. This species is phylogenetically distinct from the other Sporothrix species, based on phylogenetic analysis of combined TUB2 and CAL sequence data (Fig. 4). Sporothrix europaea is phylogenetically closely related to S. fusiformis and S. lunata described by Aghayeva et al. (2004). However, S. europaea has smaller ascomatal bases and necks compared to S. fusiformis (86–193 μm and 257–530 μm vs. 121–273 μm and 301–1168 μm). In turn, S. europea has larger ascomatal bases (86–193 μm vs. 60–178 μm) and longer necks (257–530 μm vs. 162–700 μm) compared to S. lunata (Aghayeva et al. 2004). In addition, S. europaea has two conidial types, guttuliform to fusiform and subglobose to obovate, whereas those in cultures of S. fusiformis are only guttuliform to fusiform. Spotothrix lunata has lunate conidia flattened at one side or curved with a blunt base (Aghayeva et al. 2004). The cultures of S. europaea grow on 2% MEA between 5 °C and 35 °C, whereas S. fusiformis and S. lunata did not growth below 10 °C and above 30 °C (Aghayeva et al. 2004). Sporothrix europaea was represented by five isolates collected from Poland. It corresponds to Sporothrix sp. 4 in the study of Jankowiak et al. (2019b). Sporothrix europaea was isolated from hardwoods in association with different bark beetles. 137 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Discussion In this study, a set of ophiostomatalean isolates from Norway and Poland were shown to represent two new taxa in the Ophiostomatales (Ascomycota), Ophiostoma babimostense and Sporothrix europaea. Morphological comparisons and multigene phylogenies placed O. babimostense as an additional species within the O. ulmi species complex. The new species described herein exhibit morphological resemblance to the other members of the O. ulmi complex producing pesotumand sporothrix-like morphs. Most of the members of the complex produce ascomata with long necks and allantoid ascospores that lack sheaths (De Beer and Wingfield 2013; De Beer et al. 2022). Ophiostoma babimostense is not easily distinguished morphologically from other members of the O. ulmi species complex, especially regarding morphology of the asexual morph. However, it is different from most members of the O. ulmi complex by the lack of a sexual morph in culture. Currently no sexual morphs are known for O. hylesinum and O. australiae. The first species was described from Fraxinus excelsior L. infested by bark beetles (Aas et al. 2018), while O. australiae was isolated from wounds on Acacia mearnsii De Wild. trees in Australia (Kamgan Nkuekam et al. 2008). Twenty-one known species were included previously in the O. ulmi complex (De Beer et al. 2022). The O. ulmi complex includes species what was previously referred to as the ‘hardwood clade’ in the O. piceae complex (Harrington et al. 2001; Linnakoski et al. 2010). It was later considered as the O. quercus complex (Kamgan Nkuekam et al. 2011). Finally, the O. ulmi species complex has been defined by De Beer and Wingfield (2013). The discovery of O. babimostense known hitherto only from conifers was unexpected because members of this complex had been, so far, only found in association with a variety of bark beetles on hardwoods. Nonetheless, recently, two other members of the O. ulmi complex, namely O. borealis and O. pseudokarelicum, have also been recorded from conifer hosts in the western Carpathian (Jankowiak et al. 2017). Furthermore, O. quercus, a widespread species that primarily infects sapstain in hardwood hosts (Taerum et al. 2018), has been isolated also from conifers (Reay et al. 2005; Thwaites et al. 2005; Zhou et al. 2006; Jankowiak 2012; Jankowiak and Bilański 2013a, 2013b; Jankowiak et al. 2017). It seems that some species of the O. ulmi complex can be found in a much broader range of hosts than what is commonly considered and host specificity is not as strict for some species. Ophiostoma babimostense has been isolated from Scots pine shoots infested by an unknown species of Tomicus, probably T. piniperda (L.) or T. minor (Hart.). This species was not observed in previous studies dealing with associations of Scots pine-infesting Tomicus spp. and fungi in Poland (Jankowiak 2006, 2008; Jankowiak and Bilański 2007), but was reported as an unknown species later on (Jankowiak and Kolařík 2011; labelled in this paper as Ophiostoma sp. 1). The ecological niche of O. babimostense remains incompletely unknown. It can be assumed that it is associated with Tomicus species in Poland although this should be confirmed. Sporothrix europaea forms part of the S. gossypina complex, bringing the total number of species in the complex to 19. This species complex was introduced by De Beer et al. (2016). Members of the S. gossypina complex are most often associated with bark and ambrosia beetles, although a few species have been isolated from stained oak wood or tree wounds (Aghayeva et al. 2004; De Beer et al. 2022). 138 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Sporothrix europaea was isolated from different beetle species and host trees (Jankowiak et al. 2017, 2019b; labelled in this paper as Sporothrix sp. 4), confirming that it is not host specific. In a Polish study, S. europaea was found in association with Scolytus intricatus and Xyleborus monographus on Quercus robur, Scolytus rugulosus on Prunus domestica and Anisandrus dispar and Xyleborinus saxesenii on Fagus sylvatica (Jankowiak et al. 2019b; Suppl. material 1). In addition, a single isolate of S. europaеа was isolated from Ips cembrae on Larix decidua (Jankowiak et al. 2017). The Norwegian isolates of S. europaea were collected from Q. robur infested by A. dispar and Trypodendron domesticum (H. Solheim, unpublished data). In addition, our data show that S. fusiformis is also commonly associated with diverse species of bark beetles and host trees, including A. dispar, Dryocoetes villosus, S. intricatus, S. ratzeburgi, X. monographus, X. saxesenii and Betula pendula, Fagus sylvatica, Larix decidua, Prunus domestica and Quercus robur (Jankowiak et al. 2019b; species labelled in this paper as Sporothrix sp. 9). This fungus has previously been described by Aghayeva et al. (2004), where it was found in Azerbaijan and Austria occurring on stained wood of Castanea sativa, L. decidua, Populus nigra, Quercus petraea (Aghayeva et al. 2004). In addition, S. fusiformis was previously found in association with Ips cembrae in Austria (Aghayeva et al. 2004). Species of the S. gossypina complex exhibit a relative morphological homogeneity. Based on DNA sequence comparisons, S. europaea is closely related to S. fusiformis and S. lunata. In addition to phylogenetic differences, these species can be separated from each other, based on characteristics of their sexual and asexual morphs. The present study provides novel information about the new species from the O. ulmi complex that includes non-pathogenic species like Ophiostoma quercus (Taerum et al. 2018) or the highly aggressive pathogenic fungus O. novo-ulmi, the causal agents of Dutch Elm Disease (DED) (Brasier 1991). The results of this study have also expanded our knowledge of Sporothrix. Broadly, the results suggest that Sporothrix species from the S. gossypina complex are common and ecologically diverse members of the Ophiostomatales in hardwood ecosystems in Europe. Additional information Conflict of interest The authors have declared that no competing interests exist. Ethical statement No ethical statement was reported. Use of AI No use of AI was reported. Funding This work was supported by the Ministry of Science and Higher Education of the Republic of Poland (SUB/040013–D019). 139 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Author contributions R.J. conceived the idea and performed the fungal isolates collection, analyzed the molecular data, morphological description, wrote the manuscript; H.S. performed the fungal isolates collection, analyzed the molecular data, P. B. analyzed the molecular data, phylogeny, wrote the manuscript, F.K. morphological description. Author ORCIDs Robert Jankowiak https://orcid.org/0000-0002-2804-5396 Piotr Bilański https://orcid.org/0000-0002-2750-6699 Data availability All of the data that support the findings of this study are available in the main text or Supplementary Information. References Aas T, Solheim H, Jankowiak R, Bilański P, Hausner G (2018) Four new Ophiostoma species associated with hardwood-infesting bark beetles in Norway and Poland. Fungal Biology 122: 1142–1158. https://doi.org/10.1016/j.funbio.2018.08.001 Aghayeva DN, Wingfield MJ, De Beer ZW, Kirisits T (2004) Two new Ophiostoma species with Sporothrix anamorphs from Austria and Azerbaijan. Mycologia 96: 866–878. https://doi.org/10.1080/15572536.2005.11832933 Bhunjun CS, Chen YJ, Phukhamsakda C, Boekhout T, Groenewald JZ, McKenzie EHC, Francisco EC, Frisvad JC, Groenewald M, Hurdeal VG, Luangsa-ard J, Perrone G, Visagie CM, Bai FY, Błaszkowski J, Braun U, de Souza FA, de Queiroz MB, Dutta AK, Gonkhom D, Goto BT, Guarnaccia V, Hagen F, Houbraken J, Lachance MA, Li JJ, Luo KY, Magurno F, Mongkolsamrit S, Robert V, Roy N, Tibpromma S, Wanasinghe DN, Wang DQ, Wei DP, Zhao CL, Aiphuk W, Ajayi-Oyetunde O, Arantes TD, Araujo JC, Begerow D, Bakhshi M, Barbosa RN, Behrens FH, Bensch K, Bezerra JDP, Bilański P, Bradley CA, Bubner B, Burgess TI, Buyck B, Čadež N, Cai L, Calaça FJS, Campbell LJ, Chaverri P, Chen YY, Chethana KWT, Coetzee B, Costa MM, Chen Q, Custódio FA, Dai YC, Damm U, de Azevedo Santiago ALCM, De Miccolis Angelini RM, Dijksterhuis J, Dissanayake AJ, Doilom M, Dong W, Alvarez-Duarte E, Fischer M, Gajanayake AJ, Gené J, Gomdola D, Gomes AAM, Hausner G, He MQ, Hou L, Iturrieta-González I, Jami F, Jankowiak R, Jayawardena RS, Kandemir H, Kiss L, Kobmoo N, Kowalski T, Landi L, Lin CG, Liu JK, Liu XB, Loizides M, Luangharn T, Maharachchikumbura SSN, Makhathini Mkhwanazi GJ, Manawasinghe IS, Marin-Felix Y, McTaggart AR, Moreau PA, Morozova OV, Mostert L, Osiewacz HD, Pem D, Phookamsak R, Pollastro S, Pordel A, Poyntner C, Phillips AJL, Phonemany M, Promputtha I, Rathnayaka AR, Rodrigues AM, Romanazzi G, Rothmann L, Salgado-Salazar C, Sandoval-Denis M, Saupe SJ, Scholler M, Scott P, Shivas RG, Silar P, Souza-Motta CM, Silva-Filho AGS, Spies CFJ, Stchigel AM, Sterflinger K, Summerbell RC, Svetasheva TY, Takamatsu S, Theelen B, Theodoro RC, Thines M, Thongklang N, Torres R, Turchetti B, van den Brule T, Wang XW, Wartchow F, Welti S, Wijesinghe SN, Wu F, Xu R, Yang ZL, Yilmaz N, Yurkov A, Zhao L, Zhao RL, Zhou N, Hyde KD, Crous PW (2024) What are the 100 most cited fungal genera? Studies in Mycology 108: 1–411. https://doi.org/10.3114/sim.2024.108.01 Bilański P, Jankowiak R, Solheim H, Fortuna P, Chyrzyński Ł, Warzecha P, Taerum SJ (2023) Soil-borne Ophiostomatales species (Sordariomycetes, Ascomycota) in 140 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland beech, oak, pine, and spruce stands in Poland with descriptions of Sporothrix roztoczensis sp. nov., S. silvicola sp. nov., and S. tumida sp. nov. MycoKeys 97: 41–69. https://doi.org/10.3897/mycokeys.97.97416 Błońska E, Lasota J, Jankowiak R, Michalcewicz J, Wojas T, Zbyryt A (2021) Biological and physicochemical properties of the nests of White Stork Ciconia ciconia reveal soil entirely formed, modified and maintained by birds. The Science of the Total Environment 763: 43020. https://doi.org/10.1016/j.scitotenv.2020.143020 Brasier CM (1991) Ophiostoma novo-ulmi sp. nov., causative agent of current Dutch elm disease pandemics. Mycopathologia 115: 151–161. https://doi.org/10.1007/ BF00462219 Carbone I, Kohn LM (1999) A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91: 553–556. https://doi.org/10.2307/3761358 Constantinescu O, Ryman S (1989) A new Ophiostoma on polypores. Mycotaxon 34: 637–642. https://doi.org/10.5962/p.416750 Crous PW, Costa MM, Kandemir H, Vermaas M, Vu D, Zhao L, Arumugam E, Flakus A, Jurjević Ž, Kaliyaperumal M, Mahadevakumar S, Murugadoss R, Shivas RG, Tan YP, Wingfield MJ, Abell SE, Marney TS, Danteswari C, Darmostuk V, Denchev C, Denchev TT, Etayo J, Gené J, Gunaseelan S, Hubka V, Illescas T, Jansen GM, Kezo K, Kumar S, Larsson E, Mufeeda KT, Piątek M, Rodriguez-Flakus P, Sarma PVSRN, Stryjak-Bogacka M, Torres-Garcia D, Vauras J, Acal DA, Akulov A, Alhudaib K, Asif M, Balashov S, Baral H-O, Baturo-Cieniewska A, Begerow D, Beja-Pereira A, Bianchinotti MV, Bilański P, Chandranayaka S, Chellappan N, Cowan DA, Custódio FA, Czachura P, Delgado G, Desilva NI, Dijksterhuis J, Dueñas M, Eisvand P, Fachada V, Fournier J, Fritsche Y, Fuljer F, Ganga KGG, Guerra MP, Hansen K, Hywel-Jones N, Ismail AM, Jacobs CR, Jankowiak R, Karich A, Kemler M, Kisło K, Klofac W, Krisai-Greilhuber I, Latha KPD, Lebeuf R, Lopes ME, Lumyong S, Maciá-Vicente JG, Maggs-Kölling G, Magistà D, Manimohan P, Martín MP, Mazur E, Mehrabi-Koushki M, Miller AN, Mombert A, Ossowska EA, Patejuk K, Pereira OL, Piskorski S, Plaza M, Podile AR, Polhorský A, Pusz W, Raza M, Ruszkiewicz-Michalska M, Saba M, Sánchez RM, Singh R, Liwa L, Smith ME, Stefenon VM, Strašiftáková D, Suwannarach N, Szczepańska K, Telleria MT, Tennakoon DS, Thines M, Thorn RG, Urbaniak J, Vandervegte M, Vasan V, Vila-Viçosa C, Voglmayr H, Wrzosek M, Zappelini J, Groenewald JZ (2023) Fungal Planet description sheets: 1550–1613. Persoonia 51: 280–417. https://doi.org/10.3767/persoonia.2023.51.08 Crous PW, Jurjević Ž, Balashov S, De la Peña-Lastra S, Mateos A, Pinruan U, Rigueiro-Rodrguez A, Osieck ER, Altés A, Czachura P, Esteve-Raventós F, Gunaseelan S, Kaliyaperumal M, Larsson E, Luangsa-ard JJ, Moreno G, Pancorbo F, Piątek M, Sommai S, Somrithipol S, Asif M, Delgado G, Flakus A, Illescas T, Kezo K, Khamsuntorn P, Kubátová A, Labuda R, Lavoise C, Lebel T, Lueangjaroenkit P, Maciá-Vicente JG, Paz A, Saba M, Shivas RG, Tan YP, Wingfield MJ, Aas T, Abramczyk B, Ainsworth AM, Akulov A, Alvarado P, Armada F, Assyov B, Avchar R, Avesani M, Bezerra JL, Bhat JD, Bilański P, Bily DS, Boccardo F, Bozok F, Campos JC, Chaimongkol S, Chellappan N, Costa MM, Dalecká M, Darmostuk V, Daskalopoulos V, Dearnaley J, Dentinger BTM, De Silva NI, Dhotre D, Carlavilla JR, Doungsa-ard C, Dovana F, Erhard A, Ferro LO, Gallegos SC, Giles CE, Gore G, Gorfer M, Guard FE, Hanson S-Å, Haridev P, Jankowiak R, Jeffers SN, Kandemir H, Karich A, Kisło K, Kiss L, Krisai-Greilhuber I, Latha KPD, Lorenzini M, Lumyong S, Manimohan P, Manjón JL, Maula F, Mazur E, Mesquita NLS, Młynek K, Mongkolsamrit S, Morán P, Murugadoss R, Nagarajan M, Nalumpang S, Noisripoom W, Nosalj S, Novaes Q, Nowak M, Pawłowska J, Peiger M, Pereira OL, Pinto A, Plaza M, Polemis E, Polhorský A, Ramos DO, Raza M, Rivas-Ferreiro M, Rodriguez-Flakus P, 141 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Ruszkiewicz-Michalska M, Sánchez A, Santos A, Schüller A, Scott PA, Şen İ, Shelke D, Śliwa L, Solheim H, Sonawane H, Strašiftáková D, Stryjak-Bogacka M, Sudsanguan M, Suwannarach N, Suz LM, Syme K, Taşkn H, Tennakoon DS, Tomka P, Vaghef N, Vasan V, Vauras J, Wiktorowicz D, Villarreal M, Vizzini A, Wrzosek M, Yang X, Yingkunchao W, Zapparoli G, Zervakis GI, Groenewald JZ (2024) Fungal Planet description sheets: 1614–1696. Fungal Systematics and Evolution 13: 183–440. https://doi. org/10.3114/fuse.2024.13.11 Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: More models, new heuristics and parallel computing. Nature Methods 9: 772. https://doi.org/10.1038/ nmeth.2109 De Beer ZW, Wingfield MJ (2013) Emerging lineages in the Ophiostomatales. In: Seifert KA, De Beer ZW, Wingfield MJ (Eds) The Ophiostomatoid Fungi: Expanding Frontiers. Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands, CBS Biodiversity Series 12, 21–46. De Beer ZW, Duong TA, Wingfield MJ (2016) The divorce of Sporothrix and Ophiostoma: Solution to a problematic relationship. Studies in Mycology 83: 165–191. https://doi. ogr/10.1016/j.simyco.2016.07.001 De Beer ZW, Procter M, Wingfield MJ, Marincowitz S, Duong TA (2022) Generic boundaries in the Ophiostomatales reconsidered and revised. Studies in Mycology 101: 57–120. https://doi.org/10.3114/sim.2022.101.02 De Errasti A, De Beer ZW, Coetzee MPA, Roux J, Rajchenberg M, Wingfield MJ (2016) Three new species of Ophiostomatales from Nothofagus in Patagonia. Mycological Progress 15: 17. https://doi.org/10.1007/s11557-016-1158-z De Meyer EM, De Beer ZW, Summerbell RC, Moharram AM, De Hoog GS, Vismer HF, Wingfield MJ (2008) Taxonomy and phylogeny of new woodand soil-inhabiting Sporothrix species in the Ophiostoma stenoceras–Sporothrix schenckii complex. Mycologia 100: 647–661. https://doi.org/10.3852/07-157R Feau N, Tanney JB, Herath P, Zeglen S, Hamelin RC (2024) Ophiostoma haidaense, sp. nov., a new member of the O. piceae species complex associated with yellow-cedar, Callitropsis nootkatensis. Mycologia 116(5): 694–707. https://doi.org/10.1080/0027 5514.2024.2357968 Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes: Application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113– 118. https://doi.org/10.1111/j.1365-294x.1993.tb00005.x Glass NL, Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61: 1323–1330. https://doi.org/10.1128/aem.61.4.1323-1330.1995 Grobbelaar JW, Aghayeva DN, De Beer ZW, Bloomer P, Wingfield MJ, Wingfield BD (2009) Delimitation of Ophiostoma quercus and its synonyms using multiple gene phylogenies. Mycological Progress 8: 221–236. https://doi.org/10.1007/s11557-009-0594-4 Guindon S, Gascuel O (2003) A simple, fast and accurate method to estimate large phylogenies by maximum-likelihood. Systematic Biology 52: 696–704. https://doi. org/10.1080/10635150390235520 Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology 59: 307–321. https://doi.org/10.1093/ sysbio/syq010 Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium 41: 95–98. 142 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Harrington TC, McNew D, Steimel J, Hofstra D, Farrell R (2001) Phylogeny and taxonomy of the Ophiostoma piceae complex and the Dutch elm disease fungi. Mycologia 93: 111–136. https://doi.org/10.1080/00275514.2001.12061284 Jacobs K, Bergdahl DR, Wingfield MJ, Halik S, Seifert KA, Bright DE, Wingfield BD (2004) Leptographium wingfieldii introduced into North America and found associated with exotic Tomicus piniperda and native bark beetles. Mycological Research 108: 411– 418. https://doi.org/10.1017/S0953756204009748 Jankowiak R (2006) Fungi associated with Tomicus piniperda in Poland and assessing their virulence using Scots pine seedlings. Annals of Forest Science 63: 801–808. https://doi.org/10.1051/forest:2006063 Jankowiak R (2008) Fungi associated with Tomicus minor on Pinus sylvetris in Poland and their succession into the sapwood of beetle-infested windblown trees. Canadian Journal of Forest Research 38: 2579–2588. https://doi.org/10.1139/X08-101 Jankowiak R (2012) Ophiostomatoid fungi associated with Ips sexdentatus on Pinus sylvestris in Poland. Dendrobiology (Poznań) 68: 43–54. Jankowiak R, Bilański P (2007) Fungal flora associated with Tomicus piniperda L. in an area close to a timber yard in southern Poland. Journal of Applied Entomology 131: 579–584. https://doi.org/10.1111/j.1439-0418.2007.01194.x Jankowiak R, Bilański P (2013a) Association of the pine-infesting Pissodes species with ophiostomatoid fungi in Poland. European Journal of Forest Research 132: 523–534. https://doi.org/10.1007/s10342-013-0693-2 Jankowiak R, Bilański P (2013b) Ophiostomatoid fungi associated with root-feeding bark beetles in Poland. Forest Pathology 43: 422–428. https://doi.org/10.1111/efp.12049 Jankowiak R, Kolařík M (2011) Ophiostomatoid fungi isolated from fallen shoots of Scots pine pruned by Tomicus species in Poland. Acta Mycologica 46(2): 201–210. https://doi.org/10.5586/am.2011.013 Jankowiak R, Strzałka B, Bilański P, Kacprzyk M, a Lukášová K, Linnakoski R, Matwiejczuk S, Misztela L, Rossa R (2017) Diversity of Ophiostomatales species associated with conifer-infesting beetles in the Western Carpathians. European Journal of Forest Research 136: 939–956. https://doi.org/10.1007/s10342-017-1081-0 Jankowiak R, Ostafińska A, Truls A, Solheim H, Bilański P, Linnakoski R, Hausner G (2018a) Three new Leptographium spp. (Ophiostomatales) infecting hardwood trees in Norway and Poland. Antonie van Leeuwenhoek Journal of General and Molecular Microbiology 111: 2323–2347. https://doi.org/10.1007/s10482-018-1123-8 Jankowiak R, Bilański P, Chyrzyński P, Strzałka B (2018b) Identification of sapstain fungi from Scots pine pallets and assessment of their staining ability. European Journal of Plant Pathology 150: 307–322. https://doi.org/10.1007/s10658-017-1279-5 Jankowiak R, Bilański P, Strzałka B, Linnakoski R, Bosak A, Hausner G (2019a) Four new Ophiostoma species associated with coniferand hardwood-infesting bark and ambrosia beetles from Czech Republic and Poland. Antonie van Leeuwenhoek Journal of General and Molecular Microbiology 112: 1501–15021. https://doi.org/10.1007/ s10482-019-01277-5 Jankowiak R, Strzałka B, Bilański P, Kacprzyk M, Wieczorek P, Linnakoski R (2019b) Ophiostomatoid fungi associated with hardwood-infesting bark and ambrosia beetles in Poland: Taxonomic diversity and vector specificity. Fungal Ecology 39: 152– 167. https://doi.org/10.1016/j.funeco.2019.02.001 Jankowiak R, Bilański P, Ostafińska A, Linnakoski R (2019c) Ophiostomatales associated with wounds on hardwood trees in Poland. Plant Pathology 68: 1407–1424. https://doi.org/10.1111/ppa.13061 143 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Jankowiak R, Solheim H, Bilański P, Marincowitz S, Wingfield MJ (2020) Seven new species of Graphilbum from conifers in Norway, Poland, and Russia. Mycologia 112(6): 1240–1262. https://doi.org/10.1080/00275514.2020.1778375 Jankowiak R, Szewczyk G, Bilański P, Jazłowiecka D, Harabin B, Linnakoski R (2021) Blue-stain fungi isolated from freshly felled Scots pine logs in Poland, including Leptographium sosnaicola sp. nov. Forest Pathology 51(2): e12672. https://doi. org/10.1111/efp.12672 Jankowiak R, Solheim H, Bilański P, Mukhopadhyay J, Hausner G (2022) Ceratocystiopsis spp. associated with pineand spruce-infesting bark beetles in Norway. Mycological Progress 21: 61. https://doi.org/10.1007/s11557-022-01808-x Jankowiak R, Gumulak N, Bilański P, Solheim H, Tomalak M, Wingfield MJ (2023a) Five new Graphium species from hardwood trees in Poland. Mycologia. https://doi.org/10 .1080/00275514.2023.2186676 Kamgan Nkuekam G, Jacobs K, de Beer ZW, Wingfield MJ, Roux J (2008) Pesotum australi sp. nov. and Ophiostoma quercus associated with Acacia mearnsii trees in Australia and Uganda, respectively. Australasian Plant Pathology 37: 406–416. https:// doi.org/10.1071/AP08027 Kamgan Nkuekam G, De Beer ZW, Wingfield MJ, Mohammed C, Carnegie AJ, Pegg GS, Roux J (2011) Ophiostoma species (Ophiostomatales, Ascomycota), including two new taxa on eucalypts in Australia. Australian Journal of Botany 59: 283–297. https://doi.org/10.1071/BT10231 Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7, improvements in performance and usability. Molecular Biology and Evolution 30: 772–780. https://doi.org/10.1093/molbev/mst010 Kornerup A, Wanscher JH (1978) Methuen Handbook of Colour. Third Edition. Eyre Methuen, London, 1–252. Linnakoski R, De Beer ZW, Ahtiainen J, Sidorov E, Niemelä P, Pappinen A, Wingfield MJ (2010) Ophiostoma spp. associated with pineand spruce-infesting bark beetles in Finland and Russia. Persoonia 25: 72–93. https://doi.org/10.3767/003158510X550845 Linnakoski R, Jankowiak R, Villari C, Kirisits T, Solheim H, de Beer ZW, Wingfield MJ (2016) The Ophiostoma clavatum species complex: A newly defined group in the Ophiostomatales including three novel taxa. Antonie van Leeuwenhoek Journal of General and Molecular Microbiology 109(7): 987–1018. https://doi.org/10.1007/ s10482-016-0700-y López-Romero E, Reyes-Montes Mdel R, Pérez-Torres A, Ruiz-Baca E, Villagómez-Castro JC, Mora-Montes HM, Flores-Carreón A, Toriello C (2011) Sporothrix schenckii complex and sporotrichosis, an emerging health problem. Future Microbiology 6(1): 85–102. https://doi.org/10.2217/fmb.10.157 Májek T, Kolařík M, Milenković I, Kašák J, Foit J, Tomšovský M (2025) New Ophiostoma species associated with Dryocoetes himalayensis and decline of Juglans regia in Czechia. Journal of Plant Pathology 107: 407–419. https://doi.org/10.1007/s42161024-01769-y Marincowitz S, Duong TA, De Beer ZW, Wingfield MJ (2015) Cornuvesica: A little known mycophilic genus with a unique biology and unexpected new species. Fungal Biology 119(7): 615–630. https://doi.org/10.1016/j.funbio.2015.03.007 Marmolejo JG, Butin H (1990) New conifer-inhabiting species of Ophiostoma and Ceratocystiopsis (Ascomycetes, Microascales) from Mexico. Sydowia 42: 193–199. O’Donnell K, Kistler HC, Cigelnik E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing Panama disease of banana: Concordant evidence from nucle- 144 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland ar and mitochondrial gene genealogies. Proceedings of the National Academy of Sciences of the United States of America 95: 2044–2049. https://doi.org/10.1073/ pnas.95.5.2044 O’Donnell K, Nirenberg H, Aoki T, Cigelnik E (2000) A multigene phylogeny of the Gibberella fujikuroi species complex: Detection of additional phylogenetically distinct species. Mycoscience 41: 61–78. https://doi.org/10.1007/BF02464387 Ostafińska A, Jankowiak R, Bilański P, Solheim H, Wingfield MJ (2021) Six new species of Sporothrix from hardwood trees in Poland. MycoKeys 82: 1–32. https://doi. org/10.3897/mycokeys.82.66603 Rambaut A, Drummond AJ (2007) Tracer v1.4. http://beast.bio.ed.ac.uk/Tracer Reay SD, Thwaites JM, Farrell RL (2005) A survey of Ophiostoma species vectored by Hylastes ater to pine seedlings in New Zealand. Forest Pathology 35: 105–113. https:// doi.org/10.1111/j.1439-0329.2004.00393.x Roets F, Wingfield MJ, Crous PW, Dreyer LL (2013) Taxonomy and ecology of ophiostomatoid fungi associated with Protea infructescences. In: Seifert KA, De Beer ZW, Wingfield MJ (Eds) The Ophiostomatoid Fungi: Expanding Frontiers. Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands, CBS Biodiversity Series 12, 177– 187. Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics (Oxford, England) 19: 1572–1574. https://doi. org/10.1093/bioinformatics/btg180 Seifert KA (1993) Sapstain of commercial lumber by species of Ophiostoma and Ceratocystis. In: Wingfield MJ, Seifert KA, Webber JF (Eds) Ceratocystis and Ophiostoma: taxonomy, ecology, and pathogenicity. APS Press, 141–151. Six DL (2012) Ecological and evolutionary determinants of bark beetle fungus symbioses. Insects 3: 339–366. https://doi.org/10.3390/insects3010339 Swofford DL (2003) PAUP* 4.0: phylogenetic analysis using parsimony (*and other methods). Sinauer Associates, Sunderland, Massachusetts. Taerum SJ, De Beer ZW, Marincowitz S, Jankowiak R, Wingfield MJ (2018) Ophiostoma quercus: An unusually diverse and globally widespread tree-infecting fungus. Fungal Biology 122: 900–910. https://doi.org/10.1016/j.funbio.2018.05.005 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 12: 2725– 2729. https://doi.org/10.1093/molbev/mst197 Thwaites JM, Farrell RL, Duncan SM, Reay SD, Blanchette RA, Hadar E, Hadar Y, Harrington TC, McNew D (2005) Survey of potential sapstain fungi on Pinus radiata in New Zealand. New Zealand Journal of Botany 43: 653–663. https://doi.org/10.1080 /0028825X.2005.9512982 Uzunović A, Byrne T (2013) Wood market issues relating to blue-stain caused by ophiostomatoid fungi in Canada. In: Seifert KA, De Beer ZW, Wingfield MJ (Eds) The Ophiostomatoid Fungi: Expanding Frontiers. Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands, CBS Biodiversity Series 12, 201–212. Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990 Wang HM, Wang Z, Liu F, Wu CX, Zhang SF, Kong XB, Decock C, Lu Q, Zhang Z (2019) Differential patterns of ophiostomatoid fungal communities associated with three sympatric Tomicus species infesting pines in south-western China, with a description of four new species. MycoKeys 50: 93–133. https://doi.org/10.3897/mycokeys.50.32653 145 MycoKeys 123: 121–145 (2025), DOI: 10.3897/mycokeys.123.155588 Robert Jankowiak et al.: Two new ophiostomatalean species from Norway and Poland Wang Z, Liang L, Wang H, Decock C, Lu Q (2024) Ophiostomatoid fungi associated with Ips bark beetles in China. Fungal Diversity 129: 283–364. https://doi.org/10.1007/ s13225-024-00546-7 White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (Eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1 Wingfield MJ, Seifert KA, Webber JF (1993) Ceratocystis and Ophiostoma: taxonomy, ecology, and pathogenicity. American Phytopathological Society, St. Paul, Minnesota, USA. Zhang Y, Hagen F, Stielow B, Rodrigues AM, Samerpitak K, Zhou X, Feng P, Yang L, Chen M, Deng S, Li S, Liao W, Li R, Li F, Meis JF, Guarro J, Teixeira M, Al-Zahrani HS, Pires De Camargo Z, Zhang L, de Hoog GS (2015) Phylogeography and evolutionary patterns in Sporothrix spanning more than 14 000 human and animal case reports. Persoonia 35: 1–20. https://doi.org/10.3767/003158515X687416 Zhou XD, De Beer ZW, Wingfeld MJ (2006) DNA sequence comparisons of Ophiostoma spp., including Ophiostoma aurorae sp. nov., associated with pine bark beetles in South Africa. Studies in Mycology 55: 269–277. https://doi.org/10.3767/003158510X551097 Supplementary material 1 Supplementary figures Authors: Robert Jankowiak, Halvor Solheim, Piotr Bilański, Filip Kawa Data type: docx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/mycokeys.123.155588.suppl1