Introduction

The Genera of Fungi project (http://www.GeneraOfFungi.org; Crous et al. 2014a) aims to revise the generic names of fungi that are currently accepted (Kirk et al. 2013) and anticipated to obtain ‘protected status’ at the next International Botanical Congress (IBC XIX) in China in 2017. Because the majority of fungal genera were described before the DNA era, their type species need to be recollected, epi- or neotypes designated and assigned an MBT number to ensure traceability of the nomenclatural act (Robert et al. 2013), and sequence data obtained to resolve their true higher order phylogeny. Furthermore, to move to a single nomenclature for fungi (Crous et al. 2015a), their sexual–asexual links also need to be confirmed.

Materials and methods

Isolates

Several genera were re-described based on cultures obtained from the Westerdijk Fungal Biodiversity Centre in Utrecht, The Netherlands (formerly known as CBS-KNAW) and the working collection of P.W. Crous (CPC), housed at the CBS. For fresh collections, leaves and twigs were placed in damp chambers and incubated at room temperature for 1–2 days. Single conidial or ascospore colonies were established from sporulating conidiomata or ascomata in Petri dishes containing 2% malt extract agar (MEA), as described previously (Crous et al. 1991). In the case of the dung samples, small pieces were placed on moist sterile filter paper on the lid of Petri dishes containing MEA, until the spores were shot onto the surface of the agar. After 1–2 days, single spores were picked up and transferred to fresh MEA plates. Colonies were sub-cultured onto 2% potato dextrose agar (PDA), oatmeal agar (OA), MEA (Crous et al. 2009c), autoclaved pine needles on 2% tap water agar (PNA) (Smith et al. 1996) and incubated at 25 °C under continuous near-ultraviolet light to promote sporulation. Reference strains and specimens are maintained at the CBS and CPC.

DNA isolation, amplification and analyses

Genomic DNA was extracted from fungal colonies growing on MEA using the UltraClean™ Microbial DNA Isolation Kit (MO BIO Laboratories, Inc., Solana Beach, CA, USA), according to the manufacturer’s protocol. The primers V9G (de Hoog and Gerrits van den Ende 1998) or ITS5 (White et al. 1990) and LR5 (Vilgalys and Hester 1990) were used to amplify part (ITS) of the nuclear rDNA operon spanning the 3′ end of the 18S nrRNA gene, the first internal transcribed spacer (ITS1), the 5.8S nrRNA gene, the second ITS region (ITS2) and approximately 900 bp of the 5′ end of the 28S nrRNA gene. The primers ITS4 (White et al. 1990) and LROR (Vilgalys and Hester 1990) were used as internal sequence primers to ensure good quality sequences over the entire length of the amplicon. Amplification conditions followed Cheewangkoon et al. (2008). The program SeqMan v.7.0.0 (DNASTAR, Madison, WI, USA) was used to obtain consensus sequences of each isolate. BLAST searches using ITS and LSU sequences were performed for each isolate and the closest matches were retrieved from GenBank and included in the phylogenetic analysis. The multiple sequence alignments and subsequent phylogenetic analyses of the LSU data were carried out using ClustalW and maximum likelihood (ML), respectively, under MEGA v.6.06 (Tamura et al. 2013). Sequence data were deposited in GenBank (Table 1) and the alignments and trees in TreeBASE (http://www.treebase.org). In addition, 186 LSU sequences retrieved from GenBank were included in the phylogenetic analyses (Table 2).

Table 1 Details of the strains included in this study
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Table 2 Taxa used in the phylogenetic analyses, along with their GenBank accession numbers
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Morphology

Slide preparations were mounted in lactic acid or Shear’s mounting fluid from colonies sporulating on the media previously mentioned. Sections of fruiting bodies were made by hand. Observations were made with a Zeiss V20 Discovery stereo-microscope (Zeiss, Oberkochen, Germany) and with a Zeiss Axio Imager 2 light microscope using differential interference contrast (DIC) illumination and a Nikon DS-Ri2 camera and software. Colony characters and pigment production were noted after 1 month of growth on MEA and OA (Crous et al. 2009c) incubated at 25 °C. Colony colours (surface and reverse) were rated according to the colour charts of Rayner (1970). Taxonomic novelties and new typifications were deposited in MycoBank (http://www.MycoBank.org; Crous et al. 2004).

Results

Phylogeny

The LSU alignment was used to resolve the generic placement of strains (Figs. 1, 2, 3 and 4) and the ITS (not shown) was used for species identification. According to the BLAST search results using the LSU and ITS sequences of Blastacervulus eucalypti, it was closely related with members of Asterinales (Dothideomycetes). Thus, the currently accepted members of this order as well as other orders from Dothideomycetes were included in our analysis. The final alignment contained 62 taxa including the outgroup, and 862 characters including alignment gaps; 338 of these were phylogenetically informative, 40 were variable and phylogenetically-uninformative, and 454 were constant. Tamura–Nei with Gamma distribution (TN93 + G) was used as the best-fit nucleotide substitution model for the ML analysis. The single isolate of Blastacervulus eucalypti was clustered in a well-supported clade (99% BS) together with Aulographina eucalypti and species of Alysidiella, being distinct from those species. This clade was phylogenetically related [98% bootstrap support (BS) value] with other members of Asterinaceae (Guatimosim et al. 2015) and all of them were grouped within the Asterinales clade (Fig. 1).

Fig. 1
figure 1

Maximum composite likelihood tree based on the analysis from the partial LSU sequences from species included in different orders from Dothideomycetes. Bootstrap support values above 70% are shown at the nodes. Clades are delineated based on the phylogeny of Guatimosim et al. (2015). The tree was rooted with Saccharomyces cerevisiae

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Fig. 2
figure 2

Maximum composite likelihood tree based on the analysis from the partial LSU sequences from species included in different families from Xylariales. Bootstrap support values above 70% are shown at the nodes. Species from which their LSU sequences were generated here are shown in bold. The tree was rooted with Diaporthe padi and D. perjuncta (Diaporthales)

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Fig. 3
figure 3

Maximum composite likelihood tree based on the analysis from the partial LSU sequences from species included in different families from Pleosporales. Bootstrap support values above 70% are shown at the nodes. Clades are delineated based on the phylogeny of Tanaka et al. (2015). Species from which their LSU sequences were generated here are shown in bold. The tree was rooted with Neoscytalidium novaehollandiae and N. dimidiatum (Botryosphaeriales)

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Fig. 4
figure 4

Maximum composite likelihood tree based on the analysis from the partial LSU sequences from species included in the Ophiostoma and Sporothrix clades from the Ophiostomatales according to De Beer et al. (2016) and van der Linde et al. (2016). Bootstrap support values above 70% are shown at the nodes. Species from which their LSU sequences were generated here are shown in bold. The tree was rooted with Ceratocystiopsis species (Ophiostomatales)

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The BLAST search result using the LSU and ITS regions of the five isolates of Clypeophysalospora latitans showed them to be the closest taxa members of Xylariales and, therefore, representatives of different families if this order were added to the alignment. The final alignment included 42 taxa, including the outgroup sequences. The alignment comprised 794 bp, including alignment gaps; 179 of these were phylogenetically informative, 195 were variable and phylogenetically-uninformative, and 598 were conserved. Kimura 2-parameter with Gamma distribution (K2 + G) was used as the best-fit nucleotide substitution model for the ML analysis. The five isolates of Clypeophysalospora were placed in a fully supported monophyletic clade (100% BS), phylogenetically related with the type species of Plectosphaera eucalypti, Neophysalospora eucalypti and species of Bagadiella (Fig. 2). The clade containing these genera represents a new lineage within the Xylariales and, therefore, a new family is proposed below to accommodate them.

The analysis of the LSU and ITS sequences from the five isolates of Neohendersonia kickxii using BLAST searches showed them to be the closest matches to members of Pleosporales (Dothideomycetes) but without resolution at the family level. The manually adjusted LSU alignment was constructed with representatives of different families of that order and contained 75 sequences (including the outgroup sequences) and 809 characters, including alignment gaps. In total, 224 bp were phylogenetically informative, 252 bp were variable and phylogenetically-uninformative, and 548 bp were constant. The Kimura 2-parameter model with Gamma distribution and invariable sites (K2 + G + I) was used as the best-fit nucleotide substitution model for the ML analysis. In this analysis, the isolates of Neohendersonia kickxii clustered in a strongly supported monophyletic clade (100% BS), representing a novel family lineage within Pleosporales (Fig. 3).

The BLAST search result using the LSU, ITS and β-tubulin regions of Spumatoria longicollis showed them to be the closest matches to members of Ophiostomataceae (Ophiostomatales) and, therefore, they were included in the phylogenetic tree (Fig. 4). The final alignment included 21 taxa, including the outgroup, and 689 characters, including alignment gaps; 69 of these were phylogenetically informative, 91 were variable and phylogenetically-uninformative, and 598 were constant. Tamura–Nei with Gamma distribution and invariable sites (TN93 + G + I) was used as the best-fit nucleotide substitution model for the ML analysis. Spumatoria longicollis was placed on a single branch, phylogenetically separated from the species included (De Beer et al. 2013, 2016; van der Linde et al. 2016).

The genera

Aleurocystis Lloyd ex G. Cunn., Trans. & Proc. Roy. Soc. N.Z. 84: 234 (1956).

Synonym: Matula Massee, J. Roy. Microscop. Soc. 4: 176 (1888).

Classification: Incertae sedis, Agaricales, Agaricomycetes.

Current generic circumscription: Basidiomata annual, as small pustules, cupulate-discoid to resupinate with raised margin, pale ochre to light pink, gelatinous when fresh, horny when dry. Hyphal system monomitic, generative hyphae thin- to thick-walled, with clamps. Lamprocystidia (metuloid cystidia) present, hymenial to subhymenial, thick-walled. Gloeocystidia present in one species. Dendrohyphidia present in some species, usually only slightly branched. Basidia clavate, with four sterigmata. Basidiospores large, subglobose to ellipsoid, walls thin, smooth, non-amyloid.

Type species: Aleurocystis hakgallae (Berk. & Broome) G. Cunn. 1956.

Aleurocystis hakgallae (Berk. & Broome) G. Cunn. [as ‘habgallae’], Trans. & Proc. Roy. Soc. N.Z. 84: 235 (1956). Fig. 5a, b

Fig. 5
figure 5

Species of Corticium, Aleurocystis and Licrostroma. a Corticium hakgallae (holotype, K). b Aleurocystis hakgallae (Gilbertson RLG-23290, CFMR). c Corticium subgiganteum (holotype, K). d Conidiomata of Licrostroma subgiganteum (Pinette RJP-7289, CFMR). e Basidiomata of Licrostroma subgiganteum (Pinette RJP-7290, CFMR). Scale bars: 2 cm

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MycoBank: MB 292367

Basionym. Corticium hakgallae Berk. & Broome [as ‘habgallae’], J. Linn. Soc., Bot. 14: 72 (1873) [1875].

Synonyms. Peniophora hakgallae (Berk. & Broome) Cooke, Grevillea 8: 20 (1879).

Matula poroniiforme (Berk. & Broome) Massee [as ‘poroniaeformis’], J. Roy. Microscop. Soc. 4: 176 (1888).

Cytidia cornea Lloyd, Mycol. Writ. 5: 656 (1917).

Aleurodiscus capensis Lloyd, Mycol. Writ. 6 (Letter 62): 930 (1920).

Cytidia hakgallae (Berk. & Broome) G.W. Martin, Lloydia 5: 160 (1942).

Aleurodiscus hakgallae (Berk. & Broome) Donk, Persoonia 1: 68 (1959).

Description: Basidiomata annual, cupulate-discoid to resupinate with raised margin, as small separate pustules, round to orbicular, about 5 mm in diam., more thickened in the middle, texture (in dried material) dense cartilaginous. Abhymenial surface more or less smooth, few hyaline hairs may be present. Hymenial surface whitish cream to pale ochre, smooth. Hyphal system monomitic, all hyphae with clamps, thin-walled in the subhymenium, thick-walled in subiculum, gelatinised in KOH, intricate and difficult to discern, 4–7 μm wide, hymenial elements with granular, oily substances as seen in KOH (large acicular crystals present on hymenial and tramal hyphae in RLG-23290). Subiculum thin, subicular hyphae branched, 3–9 μm wide. Lamprocystidia (metuloid cystidia) present in hymenium and subhymenium, thick-walled, conical, acute, coarsely encrusted in upper half, more or less ventricose and stalked in the subhymenium, slightly projecting above the hymenial layer, 50–140 × 9–14 μm, hyaline. Dendrohyphidia scattered among basidia, little-branched in the upper part, widened in the lower, unbranched part, about 40–50 × 4.5–7 μm. Basidia clavate to cylindrical, 50–80 × 12–18 μm, bearing four sterigmata. Basidiospores large, subglobose to widely ellipsoid, with a small, prominent apiculus, thin-walled, hyaline, smooth, with granular, oily substances as seen in KOH, negative in Melzer’s reagent, 16–23 × 13–16 μm. Conidia globose, thick-walled, walls 2.5–3 μm thick, smooth, negative in Melzer’s reagent, 17–20 μm in diameter.

Specimens examined: Sri Lanka, Hakgala, on dead bark of an angiosperm tree, sine dat., Thwaites 339 [K(M) 203643 – holotype]. USA, Hawaii, Bird park, HUNP, Kau District County, on Sapindus saponaria, 15 Jan. 2000, Gilbertson RLG-23290 (CFMR – reference material designated here); ex-reference material sequences, GenBank nos. KX358899 (ITS), KX358900 (LSU).

Notes: The genus Aleurocystis, typified with A. hakgallae, comprises three corticioid species with cupuloid to discoid basidiomes, lamprocystidia, monomitic hyphal system with clamped hyphae, and large, non-amyloid basidiospores. Morphologically, the genus was assumed by Ryvarden (1998) to be related to the genera Cytidia (in Corticiales) and Aleurodiscus (in Russulales). We scrutinised the bibliographic literature on the generic type to locate its original material. In his study on types of corticioid fungi described by M. J. Berkeley, Hjortstam (1989) also listed C. ‘habgallae’ as the generic type for Aleurocystis. Upon an inquiry to K herbarium, we received a material filed as Peniophora hakgallae (Fig. 5a). The material contains a handwritten name and number matching the protologue by Berkeley and Broome (1875). Therefore, we are of the opinion that K(M) 203643 represents the original material of Corticium hakgallae. No conidial stage was detected on this K(M) 203643 specimen. The specimen from CFMR (Fig. 5b) morphologically conforms the type and we could obtain ITS and LSU sequences from it.

A MegaBLAST search of the GenBank nucleotide database at NCBI (as of 25 May 2016) showed that the best three hits using the new LSU sequence were Gloeostereum incarnatum, Baeospora myosura and Xeromphalina campanella, with 94–95% identity over 100% query coverage. Using the new ITS sequence, the best hits were Tephrocybe cf. carbonaria and Lyophyllum spp., with 82–83% identity over 91% query coverage. Therefore, Aleurocystis sensu typi is shown to belong to the order Agaricales.

Authors: M. Ghobad-Nejhad and E. Langer

Blastacervulus H.J. Swart, Trans. Br. Mycol. Soc. 90: 289 (1988).

Classification: Asterinaceae, Asterinales, Dothideomycetes

Current generic circumscription: Foliicolous, plant pathogenic. Conidiomata acervular, brown, subcuticular with a single layer of brown epidermal cells; conidioma with brown hyphal threads, verruculose, septate, 3–4 μm diam., giving rise to conidia in a sympodial fashion, holoblastic. Conidia in a dry powdery mass, globose to ellipsoidal, medium brown, thick-walled, verruculose, guttulate, frequently in chains of two, at times with a thin mucoid sheath, base truncate, unthickened.

Type species: Blastacervulus eucalypti H.J. Swart 1988.

Blastacervulus eucalypti H.J. Swart, Trans. Br. Mycol. Soc. 90: 289 (1988). Fig. 6

Fig. 6
figure 6

Blastacervulus eucalypti (CBS 124759). a Leaf symptoms. b Vertical sections through conidioma c Conidiogenous cells (arrow) and conidia from leaves. d Colony sporulating on MEA. e, f Conidia in chains. Scale bars: b = 100 μm, all others = 10 μm

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MycoBank: MB 133297

Description: Leaf spots not extending through leaf lamina; circular to subcircular or irregular, 2–6 mm diam., becoming confluent, brown, raised with a yellow halo. Conidiomata acervular, up to 350 μm diam., brown, subcuticular with a single layer of brown epidermal cells, frequently arranged in a radiating pattern, or in circles, from centre of leaf spot. Conidioma with brown hyphal threads, verruculose, septate, 3–4 μm diam., giving rise to conidia in a sympodial fashion, holoblastic. Conidia in a dry powdery mass, globose to ellipsoidal, medium brown, thick-walled, verruculose, guttulate, frequently in chains of two, at times with a thin mucoid sheath, base truncate, unthickened, 2–3 μm diam., apex obtuse, (4)6–7(8) × (4)5–6 μm.

Culture characteristics: Colonies on MEA and SNA attaining 10 mm diam. after 1 month at 25 °C, black, membranous with scarce aerial mycelium.

Specimens examined: Australia, Victoria, Calder Highway near Woodend, on living leaves of Eucalyptus obliqua, 13 Oct. 1979, H.J. Swart (holotype DAR 58998); Mullion Creek, on Eucalyptus robertsonii subsp. hemisphaerica, 26 Jan. 2007, B.A. Summerell (epitype designated here, CBS H-20278, MycoBank MBT373086; culture ex-epitype CBS 124759 = CPC 13956).

Notes: Swart (1988) regarded B. eucalypti as reminiscent of the genus Staninwardia, but distinct in having thallic, 1-septate conidia with truncate, flat hila. Phylogenetically, the two genera are members of Dothideomycetes, but according to the phylogeny of Cheewangkoon et al. (2009), Staninwardia (represented by S. suttoni) is closely related to Capnodiales, and recently it was accommodated in the Extremaceae (Quaedvlieg et al. 2014). Blastacervulus, on the other hand, belongs to Asterinales. Another similar genus is the monotypic Fairmaniella (Sutton 1980), but in the case of F. eucalypti, conidia are smaller (4–7.5 × 3–4.5 μm) and ellipsoid-fusoid in shape. The LSU analysis showed that B. eucalypti belongs to Asterinaceae (Asterinales), which includes species forming foliicolous or lichenicolous, superficial, dark brown to black colonies (Hyde et al. 2013; Guatimosim et al. 2015). Blastacervulus eucalypti is closely related with Aulographina eucalypti and Alysidiella species (Fig. 1), all of them associated with target spot and chocolate spot disease on Eucalyptus, respectively (Cheewangkoon et al. 2012).

Authors: A. Giraldo and P.W. Crous

Clypeophysalosporaceae Giraldo & Crous, fam. nov.

MycoBank: MB 818514

Description: Endophytic, saprobic or plant pathogenic. Ascomata perithecial, brown or black, immersed, single, rarely in pairs, globose, sometimes with clypeus or pseudoclypeus, wall of several layers of brown and flattened cells. Asci mixed with paraphyses, cylindrical or obclavate, hyaline, unitunicate, 8-spored, with apical mechanism staining in Melzer’s reagent. Ascospores uniseriate or biseriate, unicellular, hyaline, fusoidal-ellipsoidal, commonly surrounded by a mucoid sheath. Asexual morph coelomycetes or hyphomycetes. Conidiomata pycnidial, globose, solitary to aggregated, brown. Conidiophores differentiated or reduced to conidiogenous cells (pycnidial form), forming fascicules or rosettes on leaves, or solitary on the hyphae (hyphomycetal form), cylindrical to subcylindrical, brown to pale grey-brown. Conidiogenous cells phialidic, ampulliform, lageniform or subcylindrical, terminal and intercalary, pale to medium brown, apex with flared collarette. Conidia solitary or in slimy heads, subcylindrical, curved, hyaline, obtuse apex and truncate to subtruncate base.

Type genus: Clypeophysalospora H.J. Swart. 1981.

Type species: Clypeophysalospora latitans (Sacc.) H.J. Swart.

Genera included: Bagadiella, Clypeophysalospora, Neophysalospora and Plectosphaera.

Clypeophysalospora H.J. Swart, Trans. Br. Mycol. Soc. 76: 93 (1981).

Classification: Clypeophysalosporaceae, Xylariales, Sordariomycetes.

Current generic circumscription: Ascomata perithecial, scattered, immersed, single or rarely in pairs, covered by a dark clypeus, with a wall consisting of darkened, flattened, thin-walled cells and with a narrow periphysate ostiole. Asci paraphysate, cylindrical or slightly swollen, unitunicate, with an amyloid apical ring and a pulvillus. Ascospores mostly uniseriate or irregularly arranged, hyaline, ellipsoid, with mucous outer wall layer. Asexual morph unknown.

Type species: Clypeophysalospora latitans (Sacc.) H.J. Swart 1981.

Clypeophysalospora latitans (Sacc.) H.J. Swart, Trans. Br. Mycol. Soc. 76: 95 (1981). Fig. 7

Fig. 7
figure 7

Clypeophysalospora latitans (CPC 14930). a Leaf symptoms. b, c Vertical sections through ascoma. d Ascoma on PNA. e Ascoma. f Details of ostiolar region and peridium. g Asci. h Ascospores (arrows denote mucoid sheaths). Scale bars: e = 50 μm, all others = 10 μm

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MycoBank: MB 112195

Basionym. Physalospora latitans Sacc. Flora. Myc. Lusitan. 67 (1893).

Synonym. Amerostege latitans (Sacc.) Theiss. Annales. Mycologici. 14: 411 (1917).

Description: Ascomata visible as erumpent, hemispherical, brown on the host surface; in vertical section 320–450 μm diam., immersed, globose, clypeate, with a central periphysate ostiolar canal, surface of textura epidermoidea or globosa. Peridium 13–16 μm wide, comprising several layers of brown compressed cells. Clypeus composed of dark brown compressed host cells and fungal hyphae. Paraphyses up to 6.4 μm in diam., hypha-like, flexuose, septate, numerous, hyaline and embedded in a gelatinous matrix. Asci 8-spored, cylindrical, unitunicate, thin-walled, with a J+ and an amyloid apical ring, in vivo 115–141 × 11–16 μm, in vitro (107)108–117(118) × 9–10 μm. Ascospores uniseriate, unicellular, ellipsoidal, slightly curved, smooth-walled, in vivo 13–22 × 6–9 μm, in vitro (11)13–19(20) × 5–7 μm, hyaline, surrounded by a thin mucilaginous sheath. Asexual morph not observed.

Culture characteristics: Colonies on OA and MEA attaining 10 mm diam. after 3 weeks at 25 °C, flat, floccose to granulose, dirty white to pale rose.

Specimens examined: France, Corsica, Porto-Vecchio, on living leaves of Eucalyptus sp., 2 May 2005, P. W. Crous (CPC 27352). Portugal, Algarve, Faro, on living leaves of Eucalyptus sp., 24 Jan. 2008, P. W. Crous (epitype designated here, CBS H-20289, MycoBank MBT373087; culture ex-epitype CBS 141463 = CPC 14930, CPC 14931, CPC 14932); Beira Litoral, Coímbra (PAD 2747 lectotype). South Africa, Stellenbosch Mountain, on living leaves of Eucalyptus sp., 8 Aug. 2011, P. W. Crous (CPC 19661).

Notes: After the morphological examination of fresh material of Clypeophysalospora latitans, Kang et al. (1999) considered that the presence of immersed and clypeate ascomata, among other morphological characteristics, were similar to those observed in the genus Clypeosphaeria, and, therefore, C. latitans was accommodated in Clypeosphaeriaceae together with the other 16 allied genera (Kang et al. 1999). However, no molecular data were provided at that time. Although the type species of that family is Clypeosphaeria mamillana, Senanayake et al. (2015) used the LSU and ITS sequences of C. uniseptata (the only species from the genus having living culture at that moment) to demonstrate the affinity of this family with the order Amphisphaeriales, previously considered as a family in the Xylariales. However, presently, the Amphisphaeriales is no longer accepted as a separate order, and it is treated as one of the families in Xylariales (Crous et al. 2015a; Maharachchikumbura et al. 2016). Recently, C. mamillana has been epitypified and the polyphyly of the genus has been demonstrated (Jaklitsch et al. 2016). Clypeosphaeria mamillana belongs to Xylariaceae and C. uniseptata was combined in Lepteutypa (Amphisphaeriaceae). In consequence, the concept of the family Clypeosphaeriaceae (based on C. uniseptata; Senanayake et al. 2015) is no longer valid (Jaklitsch et al. 2016).

We have included the sequence of C. uniseptata in our LSU analysis (Fig. 2) and, according to our results, this species is not related at the family level with Clypeophysalospora latitans, and, therefore, we have proposed a new family to accommodate the latter taxon. The members of the new family, i.e. Bagadiella, Clypeophysalospora, Neophysalospora and Plectosphaerella, share morphological characteristics with each other, in terms of both sexual and asexual morph, host specificity (mainly Eucalyptus spp.) and distribution (Australia, South Africa). However, in the case of N. eucalypti, it has also been isolated from Corymbia henryi (Myrtaceae) in Mozambique and C. latitans was originally described on Eucalyptus leaves from Europe (Swart 1981; Cheewangkoon et al. 2009; Crous et al. 2011, 2014c).

Authors: A. Giraldo and P.W. Crous

Licrostroma P.A. Lemke, Canad. J. Bot. 42: 762 (1964).

Synonym: Michenera Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 333 (1868) [1869].

Classification: Peniophoraceae, Russulales, Agaricomycetes.

Current generic circumscription: Basidiomata resupinate to effused-reflexed, smooth, cream to ochre, confluent, more or less membranaceous. Hyphal system dimitic, generative hyphae simple-septate, binding hyphae as ‘bovista-type’, antler-like, branched hyphae especially present in basal layers, negative in Melzer’s reagent. Dendrohyphidia present, simple and little-branched. Cystidia smooth, long cylindrical to subclavate, thin- to thick-walled, flexuous, with rounded or tapering apices, not projecting above the hymenium. Basidia large, clavate to cylindrical, with four strigmata. Basidiospores large, globose to subglobose, thick-walled, smooth, negative in Melzer’s reagent.

Conidial state (see Fig. 5d): Conidiomata cupulate, brown, rounded, ca. 1 cm wide, with yellowish cream outer surface, attached centrally to the substratum, composed of chlamydospores and simple-septate hyphae. Chlamydospores fusoid, walls brown, 3–3.5 μm thick, smooth.

Type species: Licrostroma subgiganteum (Berk.) P.A. Lemke 1964.

Licrostroma subgiganteum (Berk.) P.A. Lemke, Canad. J. Bot. 42: 763 (1964). Fig. 5d, e

MycoBank: MB 333369

Basionym. Corticium subgiganteum Berk., Grevillea 2: 3 (1873).

Synonyms. Peniophora subgigantea (Berk.) Massee, J. Linn. Soc., Bot. 25: 142 (1889).

Terana subgigantea (Berk.) Kuntze, Revis. Gen. Pl. (Leipzig) 2: 873 (1891).

Aleurodiscus subgiganteus (Berk.) Höhn., Sber. Akad. Wiss. Wien, Math.-Natur. Kl. Sitzungsb. 121: 342 (1912).

Michenera artocreas Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 333 (1868).

Aleurodiscus orientalis Lloyd, Mycol. Writ. 6 (Letter 62): 927 (1920).

Aleurodiscus reflexus Yasuda, Bot. Mag., Tokyo 35 (no. 420): 269 (1921).

Description and illustration: Lemke (1964), Ryvarden (2010).

Specimens examined. USA, South Carolina, Aiken, on Magnolia glauca, sine dat., Ravenel 1669 (K(M)203642 – holotype); Maine, Aroostook, on dead snag of Acer rubrum, 31 Oct. 2006, R. J. Pinette RJP-7289 (CFMR – reference material designated here); ex-reference material sequences, GenBank nos. KX358901, KX358903 (ITS); KX358902, KX358904 (LSU).

Notes: Licrostroma is a monotypic genus of corticioid basidiomycetes. It has thick, effused-reflexed basidiomes with compact texture, a dimitic hyphal system, simple-septate generative hyphae and branched binding-like hyphae called ‘bovista-type hyphae’ by Lemke (1964), who proposed the genus to be related to Aleurodiscus. Indeed, the ‘bovista-type’ hyphae mentioned by Lemke (1964) somehow resemble dichohyphae in Scytinostroma and asterosetae (asterohyphae) in Asterostroma. Licrostroma and Scytinostroma, as well as Asterostroma, are dimitic and have simple-septate generative hyphae (except for a few Scytinostroma species, which have clamps). However, unlike the two latter genera, Licrostroma does not react with Melzer’s reagent.

We examined two specimens at CFMR, one containing only brown conidiomata (Fig. 5d) and the other specimen with resupinate basidiomata (Fig. 5e). Both samples have been collected on the same date and substratum, and from the same locality. We obtained the ITS and LSU sequences from both specimens and believe that they represent the same species, probably a single collection divided into sexual and asexual specimens. Morphologically, the CFMR collection conforms to the original material of Corticium subgiganteum (Fig. 5c) that we received from K. We could also observe some chlamydospores within the hyphae of the K type specimen.

A MegaBLAST search of the GenBank nucleotide database at NCBI (as of 25 May 2016) showed that the top hits using the new LSU sequences of L. subgiganteum were Scytinostroma spp., Asterostroma spp. and Gloiothele spp. [all in Peniophoraceae sensu Miller et al. (2006)], with 92–94% identity over 97–100% coverage. Using the new ITS sequences, the top hits were Scytinostroma and Gloiothele, as well as hits of Helicobasidium and Tuberculina maxima (asexual morph Helicobasidium). Although we are not able to interpret the latter hits, the morphological similarities between Licrostroma and Scytinostroma/Asterostroma described above suggest their close relationships. Upon this, Licrostroma sensu typi is proposed to belong to Peniophoraceae, Russulales.

Authors: M. Ghobad-Nejhad and E. Langer

Neohendersoniaceae Giraldo & Crous, fam. nov.

MycoBank MB 818515.

Description: Conidiomata single or gregarious, immersed, globose to collabent, papillate, dark brown to black, unilocular or multilocular; wall thick, of textura porrecta, intricata or angularis. Ostiole single, papillate. Conidiophores reduced to conidiogenous cells. Conidiogenous cells discrete, determinate or indeterminate, cylindrical, lageniform, doliiform or ampulliform, hyaline, smooth, with percurrent proliferations. Conidia obovoid, cylindrical, clavate or fusiform, distoseptate or euseptate, basal or apical cells paler than the median cells, thick-walled, base truncate, apex obtuse.

Type genus: Neohendersonia Petr., 1921.

Type species: Neohendersonia kickxii (Westend.) Sutton & Pollack.

Neohendersonia Petr., Annls Mycol. 19 (3–4): 190 (1921).

Classification: Neohendersoniaceae, Pleosporales, Dothideomycetes.

Current generic circumscription: Mycelium immersed, branched, septate, pale brown. Conidiomata pycnidial, eustromatic, separate or aggregated, immersed, globose or collabent, papillate, dark brown to black, uni- or multilocular, thick-walled, the outer one dark brown with textura porrecta or intricata, and the inner one hyaline with textura angularis. Ostiole single, circular, papillate. Conidiophores reduced to conidiogenous cells. Conidiogenous cells holoblastic, single or annellidic, discrete, determinate or indeterminate, cylindrical, lageniform, doliiform or ampulliform, hyaline, smooth, with 0–3 percurrent proliferations. Conidia obovoid, cylindrical, clavate or fusiform, transversely distoseptate or euseptate, versicoloured, basal and sometimes apical cells paler than the median cells, thick-walled, smooth, truncated at base, obtuse at the apex, eguttulate.

Type species: Neohendersonia kickxii (Westend.) Sutton and Pollack 1974.

Neohendersonia kickxii (Westend.) Sutton & Pollack, Mycopath. Mycol. Appl. 52: 334 (1974). Fig. 8

Fig. 8
figure 8

Neohendersonia kickxii (CBS 112403). a Conidioma on Fagus sylvatica. b Conidiogenous cell in vivo. c Conidia in vivo showing the paler basal cell. d Conidiogenous cells in vitro (arrows). e Conidia in vitro. f Second kind of conidia growing directly from the hyphae. Scale bars: b–e = 10 μm, f = 5 μm

Full size image

MycoBank: MB 318608

Basionym. Stilbospora kickxii Westend., Bull. Acad. R. Sci. Belg., Cl. Sci. 18: 409 (1851).

Synonyms. Hendersonia loricata Sacc. & Roum., Michelia 2 (8): 629 (1882).

Coryneum kickxii (Westend.) Traverso, Bol. Soc. Bot. Ital. 215 (1904).

Neohendersonia pyriformis (G.H. Otth) Petr., Annls Mycol. 19 (3–4): 191 (1921).

Stilbospora pyriformis (G.H. Otth) Grove, British Stem- and Leaf-Fungi (Coelomycetes) (Cambridge) 2: 326 (1937).

Description: Conidiomata pycnidial, separate or aggregated, immersed, subperidermal, not protruding, black, smooth, ampulliform, uni- or multilocular, up to 1300 μm diam., outer layer dark brown, 30 μm wide, inner layer pale brown becoming hyaline towards the conidiogenous cells, 40–60 μm wide. Ostiole single, circular, up to 250 μm diam., slightly papillate, thin-walled and pale brown. Conidiogenous cells holoblastic, annellidic, with up to seven annellations, discrete, hyaline, smooth, cylindrical to obpyriform, in vivo 4–9 × 2–6 μm, in vitro 6–9 × 6–8 μm. Conidia solitary, obovoid to clavate, pyriform 2–3 distoseptate, slightly constricted at the prominent septa, apical cell larger than the other ones, thick- and smooth-walled, apex obtuse, base truncated, brown with paler basal cell, in vivo 21–45.5 × 12.5–20 μm, (2)3(6)-celled and seldom with one longitudinal septum per conidium, in vitro (20)22–26(28) × 11–14 μm. Microconidia were observed in culture growing directly from the hyphae, cylindrical with rounded apex and truncate base, hyaline, thick- and smooth-walled, 2–4 × 2–3 μm.

Culture characteristics: Colonies on MEA and OA black, flat, cottony, aerial mycelium dirty white, diffusible amber pigment, reverse honey to isabelline.

Specimens examined: Austria, Carinthia, Sankt Margareten im Rosental, on Fagus sylvatica, unknown date, W. Jaklitsch (CBS 122938); Grieskirchen, Natternbach, on Fagus sylvatica, unknown date, H. Voglmayr (CBS 122941). Belgium, Courtrai, Parc Saint-George, on branch of Fagus sylvatica (“substrate originally determined as Betula pubescens and later corrected with Fagus sylvatica”), Mar. G.D. Westendorp (holotype IMI 168433 ex BR). Germany, mixed forest, on twigs of Fagus sylvatica, 19 Apr. 2014, R.K. Schumacher (CPC 24865). Italy, Pian di Novello, on bark of twigs from Fagus sylvatica, 8 May 1996, R. Danti (epitype designated here, MycoBank MBT373088; culture ex-epitype CBS 112403). Sweden, Uppland, on Fagus sp. 12 Mar 1989, K. & L. Holm (CBS 114276 = UPSC 2890).

Notes: Neohendersonia was established by Petrak (1921) based on Neohendersonia pyriforme, described from Fagus sylvatica in Belgium. Sutton and Pollack (1974) subsequently combined Stilbospora kickxii into Neohendersonia, and N. kickxii was proposed as the type species, being the older name. After the inclusion of N. congoensis, the generic concept was expanded to include species with euseptate conidia (Sutton 1975). Recently, a third species, N. fagi, has been described from the same source in Italy (Wijayawardene et al. 2016). Currently, from the three species accepted in the genus, only N. kickxii has living type material, and, therefore, we cannot elucidate the monophyly of the genus and the species boundaries. However, the three species can be morphologically distinguished by the presence of euseptate conidia in N. congoensis and the size of the distoseptate conidia in N. fagi (30–48 × 10–15 μm) and N. kickxii (21–45.5 × 12.5–20 μm) (Sutton 1975, 1980; Wijayawardene et al. 2016).

From the material examined here, the culture CPC 24865 was found to be phylogenetically distinct from the other isolates of N. kickxii and it could represent a different taxon. However, we prefer to not assign a name to this culture yet, since we did not find sufficient morphological characteristics to propose it as a different species.

Neohendersonia kickxii has been reported as a specific endophyte of beech twigs in Europe (Danti et al. 2002; Sieber 2007) and from beech bark in North American forests (Griesmer-Zakhar 2013), while N. congoensis and N. fagi have been described on stems of Aloe or Agave in Congo and branch of Fagus in Italy, respectively.

Authors: A. Giraldo, R.K. Schumacher and P.W. Crous.

Spumatoria Massee and E.S. Salmon, Ann. Bot. Lond. 15: 350 (1901).

Classification: Ophiostomataceae, Ophiostomatales, Sordariomycetes.

Current generic circumscription: Saprobic, coprophilous. Ascomata perithecial, single, gregarious, immersed, erumpent, loculus globose with one or seldom two necks per perithecium. Ascomatal neck centrally, terete, apically paler and conspicuously long fimbriate, basally dark brown to black, felty, textura prismatica. Peridium dark brown to black, slightly hard, rough, basally with red brown hyphae, textura angularis. Paraphyses longer than the asci. Asci 8-spored, clavate, apically rounded and with an ocular chamber, pedicel mostly short and not furcate, thick-walled, bitunicate. Ascospores obliquely biseriate, overlapped, 2(−3)-celled, clavate, straight, hyaline, thin-walled, smooth, coarsely guttulate, septa smooth and thin-walled. Asexual morph sporothrix-like (in culture). Conidiophores more or less differentiated, unbranched, cylindrical. Conidiogenous cells polyblastic, sympodial, terminal, denticulate. Conidia solitary, dacrioid, hyaline. Blastoconidia growing directly from undifferentiated hyphae, lateral, globose to subglobose.

Type species: Spumatoria longicollis Massee & E.S. Salmon 1901.

Spumatoria longicollis Massee and E.S. Salmon, Ann. Bot. Lond. 15: 351 (1901). Fig. 9

Fig. 9
figure 9

Spumatoria longicollis (CBS 141464). a Ascoma. b Ostiolar hyphae. c Ascospores. d Conidiogenous cells (arrows denote the denticles). e Conidia. f Blastoconidia. Scale bars: a = 100 μm, all others = 10 μm

Full size image

MycoBank: MB 171713

Description: Ascomata single, gregarious, immersed, erumpent, loculus globose with one or seldom two necks per perithecium, up to 500 μm long × 39–60 μm wide. Ascomatal neck central, terete, apically paler and conspicuously long fimbriate, basally dark brown to black, felty, textura prismatica, up to 430 μm long × 28–30 μm wide. Peridium dark brown to black, slightly hard, rough, basally with red brown hyphae, textura angularis. Paraphyses longer than the asci. Asci dehiscent, 8-spored, clavate, apically rounded and with an ocular chamber, pedicel mostly short and not furcate, thick-walled, bitunicate. Ascospores obliquely biseriate, overlapped, 2(−3)-celled, clavate, straight, hyaline, thin-walled, smooth, coarsely guttulate, septa smooth and thin-walled, (17)18–23(26) × 4–7.5 μm. Asexual morph sporothrix-like (in culture). Conidiophores poorly differentiated, unbranched, cylindrical, up to 30 μm long. Conidiogenous cells polyblastic, sympodial, terminal, denticulate. Conidia dacrioid, hyaline, smooth- and thin-walled, 3–4.5 × 1.5–2 μm. Blastoconidia growing directly from undifferentiated hyphae, lateral, globose to subglobose, smooth- and thick-walled, light brown, 3–4 × 2–4 μm wide.

Culture characteristics: Colonies on OA attaining 13–20 mm diam. after 1 week at 25 °C, flat, scarce aerial mycelium, dark brown. On MEA flat, creamy with scarce aerial mycelium, dirty white turning dark brown with age.

Specimens examined: England, Essex, Epping Forest, horse dung, G.E. Massee & E.S. Salmon [Massee G.E and E.S. Salmon, Ann. Bot. Lond. 15, Fig. 27] (Lectotype designed here, MycoBank MBT373089). Netherlands, near Oostvoorne, dune grassland, on cow (Galloway) dung, 26 Jan. 2016, coll. J. van der Lee, det. R.K. Schumacher, iso. A. Giraldo (epitype designated here, CBS H-22665, MycoBank MBT373090; culture ex-epitype CBS 141464 = CPC 30521).

Notes: As far as we know, this is the first report of this fungus after its original description. The sexual morph was only observed in natural substrata; many attempts to obtain this morph in culture resulted only in the sporothrix-like asexual morph. Several macroscopic and microscopic pictures were observed to corroborate the identity from the specimen. In the original protologue, Massee and Salmon (1901) mentioned cylindrical asci, 110–113 × 13–15 μm, versus narrow clavate in the illustration, and 2-celled, ellipsoidal (‘oblongis utrinque rotundatis’) spores, 15–19 × 5 μm. Unfortunately, due to the dehiscent nature of the asci and the scarce material available, these structures were not observed in this study. The authors refer the perithecia at the conidium-bearing stage to the genus Rhynchophoma, a similar genus described from the wood of Tilia ulmifolia (Karsten 1884). However, the type of this fungus is apparently lost and, according to De Beer et al. (2013), this genus does not belong to either Ophiostomatales or Microascales.

Among the coprophilous fungi, Kathistes is one of the genera morphologically similar to Spumatoria in having transversely septate ascospores and perithecia with long and dark necks, but can be differentiated by its narrower ascospores, particular sporidiomata and absence of paraphyses (Malloch and Blackwell 1990). Based on their morphological similarities and common habit, they were placed together with Klasterskya in the Kathistaceae within Ophiostomatales (Hawksworth et al. 1995; Kirk et al. 2008). However, the analysis of the SSU region showed the affinity of this family to be with the Dothideomycetes rather than the Ophiostomatales (Blackwell and Spatafora 1994); consequently, Klasterskya and Spumatoria were excluded from the latter order based on the morphological characters as reported in the original description (De Beer et al. 2013). It is important to mention that, from the approximately 300 genera currently included in the Ophiostomatales, Spumatoria is the only genus showing septate ascospores. However, our results of the molecular data, which include LSU (Fig. 4), ITS and β-tubulin (data not shown) loci, demonstrate the inclusion of the latter genus within the Ophiostomatales. Based on phylogenetic placement, Spumatoria could be considered a synonym of the older name Sporothrix (Hektoen and Perkins 1900). Although S. longicollis also have a sporothrix-like asexual state, it differs from other Sporothrix spp. based on septate ascospores, light coloured ascomata and dung-inhabiting biology. We, thus, suggest that further study including sequences of multiple gene regions and more taxa is needed in order to elucidate its relation with the type species of Sporothrix, S. schenckii and other species in the genus.

Authors: RK Schumacher, A Giraldo and PW Crous