Analysis of sequence data

Due to the poor representation of other gene sequences for Xylariales, only the ITS and LSU rDNA were used for phylogenetic inference. To complement the data matrix, ITS and LSU sequences of a representative selection of Xylariales were downloaded from GenBank; for rooting the tree, LSU sequences of eight taxa of Sordariomycetes were included (Table 1). All alignments were produced with the server version of MAFFT (www.ebi.ac.uk/Tools/mafft), checked and refined using BioEdit v. 7.0.4.1 (Hall 1999). For phylogenetic analyses, the ITS and LSU matrices were combined. After exclusion of poorly aligned regions for the ITS containing large indels, the final matrix contained 518 and 852 nucleotide characters from the ITS and LSU, respectively.

Maximum likelihood (ML) analyses were performed with RAxML (Stamatakis 2006) as implemented in raxmlGUI 1.3 (Silvestro & Michalak 2012), using the ML + rapid bootstrap setting and the GTRGAMMAI substitution model with 1 000 bootstrap replicates. The matrix was partitioned for ITS and LSU, and substitution model parameters were calculated separately for the two regions.

Maximum parsimony (MP) bootstrap analysis of the combined matrix was performed with PAUP v. 4.0a142 (Swofford 2002). All molecular characters were unordered and given equal weight; analyses were performed with gaps treated as missing data; the COLLAPSE command was set to MINBRLEN. 1 000 MP bootstrap replicates were performed, with 5 rounds of random sequence addition and subsequent TBR branch swapping (MULTREES option in effect, steepest descent option not in effect), with each replicate limited to 1 million rearrangements.

Taxonomy

Families are given in alphabetical order and the treated taxa are presented under each family: Amphisphaeriaceae (Lepteutypa), Beltraniaceae (Beltraniella carolinensis), Pseudomassariaceae (Pseudapiospora, Pseudomassariella), Requienellaceae (Acro-cordiella, Requienella), Sporocadaceae (Blogiascospora/Seiridium, Hymenopleella, Strickeria), Xylariaceae (Clypeosphaeria), Xylariales inc. sed. (Basiseptospora).

Amphisphaeriaceae G. Winter 1885

Lepteutypa Petr., Ann. Mycol. 21: 276. 1923

Type species. Lepteutypa fuckelii (G.H. Otth) Petr.

Lepteutypa fuckelii (G.H. Otth) Petr., Ann. Mycol. 21: 276. 1923 — Fig. 2

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Fig. 2

Lepteutypa fuckelii. a–c. Ostioles and clypei on the bark surface; d, i. ascus apices in Lugol; e–h. asci (g, h. in Lugol); j–q. ascospores (l, n–p. in 3 % KOH; q. in Lugol) (a, b, j, k: WU 33555; c, e–h, l, m, q: WU 33554; d, i, n: G00127792; o: B700009240; p: G00127791). — Scale bars: a, c = 0.4 mm; b = 0.2 mm; d, j–m, p, q = 7 μm; e–h, n, o = 15 μm; i = 5 μm.

Basionym. Massaria fuckelii G.H. Otth, Mitth. Naturf. Ges. Bern Nr. 654–683: 50. 1868.

= Massaria fuckelii Fuckel, Jahrb. Nassauischen Vereins Naturk. 23–24: 155. 1870. [1869–70].

Typification. Neotype proposed here: BELGIUM, Leuven, Heferlee, Heferleebos, on Tilia cordata, soc. Amphisphaeria multipunctata, 31 May 2012, P. Bormans (WU 33554; MBT202953; ex-neotype culture LEF = CBS 140409).

Perithecia immersed in sometimes slightly uneven bark, only visible as minute dark dots, sometimes surrounded by grey clypei 0.3–0.6 mm diam on the otherwise unaltered, light-coloured bark surface, scattered or aggregated in small groups, depressed globose, dark, (0.25–)0.45–0.57(–0.6) mm diam, 0.2–0.4 mm high; peridium 20–40 μm thick, brown, hyaline inside, pseudoparenchymatous. Ostioles inconspicuous to blunt papillate and shiny black, 80–150 μm diam, sometimes region around ostiole slightly darker and depressed; sometimes whitish to greyish brown mycelium present around perithecia. Apically free paraphyses 2–6 μm wide, simple, multiguttulate. Asci (107–)111–155(–185) × (9.8–)10.5–13.3(–14.5) μm (n = 20), cylindrical, with 8 uniseriate ascospores, with a short stipe, croziers and a thickened apex containing a thin amyloid apical ring, (3.0–)3.2–4.2(–4.5) μm wide and (0.7–)0.8–1.2(–1.4) μm high (n = 20). Ascospores (15.5–)17.5–22.8(–29.7) × (5.7–) 6.5–8.0(–8.7) μm, l/w (2.0–)2.4–3.2(–4.2) (n = 114), oblong or narrowly fusiform, first hyaline, turning yellow to yellow-brown, with 1 median, slightly constricted euseptum and 2 distosepta, straight to slightly curved, multiguttulate when vital, hyaline to yellowish and with a narrow mucous sheath (e.g. 1.5 μm wide) or polar caps in the ascus when young, turning olive-green in Lugol, with 1 guttule per cell, massaria-like, i.e. with a dramatic difference between the living and dead state.

Culture characteristics — Growth radius after 1 mo at room temperature on CMD up to c. 25 mm, on MEA to c. 32 mm; colony circular, sometimes finely zonate, white, becoming brown, olive to nearly black from the centre, odour indistinct. No asexual morph observed.

Habitat — In bark of Tilia spp.

Distribution — Europe.

Additional material examined. GERMANY, Nordrhein-Westfalen, Schlosspark Türnich at Kerpen, MTB 5106/142, 4 Mar. 2013, on attached branches of Tilia cordata, B. Wergen (WU 33555; culture LEF1); Rheingau, on Tilia, L. Fuckel, det. Th. Nitschke (B700009241); near Kiedrich, on Tilia, in spring, L. Fuckel, Fungi Rhen. Exs. 1769 (B700009240, G00127792); Oestrich, at Schloß Reichartshausen, on Tilia, in the winter, L. Fuckel (Nassau’s flora, G00127791).

Notes — Lepteutypa fuckelii is an uncommon fungus. A good description of the species can be found in Petrak (1923). He interpreted perithecial colonies as eutypoid stromata forming small irregular spots, but noted that stromatic tissue is only comprised by subhyaline to brownish hyphae. Aberrant ascospores may contain 5 septa with sizes up to 35 × 10 μm. No asexual morph has been found for this species.

Nomenclatural background: Nitschke created the name Massaria fuckelii, but did not publish it. However, he corresponded with both Otth and Fuckel about the fungus, who collected their own materials and independently described the fungus. Otth (1868) was first, i.e. Fuckel’s Massaria fuckelii is a heterotypic homonym. Both authors cited Nitschke as the author in order to honour him as the creator of the name, but Nitschke did not provide the descriptions. Nitschke’s impact could be honoured as author citations Nitschke ex Otth and Nitschke ex Fuckel, which, however, is not necessary. Specimens of Massaria fuckelii Fuckel collected by Fuckel, which are extant in B and G and labelled Typus, are not type specimens of Massaria fuckelii G.H. Otth. As there is no type material of G.H. Otth available, we propose a neotype here.

Lepteutypa hederae (Fuckel) Rappaz, Mycol. Helv. 7: 160. 1995 — Fig. 3

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Fig. 3

Lepteutypa hederae (G00111726). a. Ascoma and clypeus in vertical section; b, c. ascus apices (b. in Lugol; c. in aqueous cotton blue); d–i. ascospores (e, f. showing ridges; f. in transverse optical section; h, i. in 3 % KOH). — Scale bars: a = 0.2 mm; b–d, i = 10 μm; e = 7 μm; f, g = 5 μm; h = 15 μm.

Basionym. Amphisphaeria hederae Fuckel, Jahrb. Nassauischen Vereins Naturk. 25–26: 304. 1871.

≡ Anthostoma hederae (Fuckel) Sacc., Syll. Fung. (Abellini) 1: 301. 1882.

Holotype. SWITZERLAND, near Neuchatel, on dead, corticated branches of Hedera helix, by xylotomy probably Viburnum sp., Mar. 1867, P. Morthier (G00111726!; Herbier Fuckel 1894 ex Herb. Barbey-Boissier, as Amphisphaeria hederae and Anthostoma hederae on the label).

Perithecia immersed in bark and wood beneath black clypei being distinct in wood but diffuse in bark in surface view, flat, convex to pulvinate, 0.7–1.2 mm diam, 80–160 μm thick; perithecia globose, 0.35–0.7 mm diam, 0.25–0.6 mm high; peridium brown, pseudoparenchymatous. Paraphyses numerous, simple. Asci oblong, containing 8 uniseriate ascospores, unitunicate but thick-walled, apex not containing a ring, inamyloid, not staining with Congo Red, a diffuse pulvillus turning blue in Cotton Blue (in water); stipe short. Ascospores (19.5–)22–29.2(–35.3) × (7.0–)9.2–12.2(–14) μm, l/w (1.6–)1.8–3.1(–4.6) (n = 40), ellipsoid, ovoid to oblong, usually inequilateral with pointed ends, 3-distoseptate, first hyaline, turning brown, indistinctly striate, appearing warted in section.

Habitat — In bark and wood of ?Viburnum sp.

Distribution — Switzerland, only known from the holotype.

Notes — See under Lepteutypa sambuci for a comparison of L. hederae with that species. The generic affiliation of L. hederae is still unverified, as no fresh material and sequence data are available.

Lepteutypa sambuci Jaklitsch & Voglmayr, sp. nov. — MycoBank MB814827; Fig. 4

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Fig. 4

Lepteutypa sambuci. a, b. Ostioles and clypei on the bark surface; c, d. stromata in face view in wood; e. stroma and ascomata in transverse section; f. ascoma in vertical section; g. ascomatal wall in section; h, i, m, s. ascus apices (h, i, s. in Lugol; h, i. immature); j–l. asci (j. immature, in Lugol; l. in 3 % KOH); n–r. ascospores (n. in Congo Red; o, r. in Lugol/KOH; p, q. in Lugol) (a, b, g, n: WU 33556 RS; c, h, p: WU 33559 (c. image by T. Richter); d, e, l, o, r: WU 33558; f, i–k, q, s: WU 33557; m: East Cornwall (k. image by P. Cannon)). — Scale bars: a, b, d, e = 0.5 mm; c = 2 mm; f = 0.2 mm; g, j–m, r = 15 μm; h = 3 μm; i, n, p, q = 7 μm; o, s = 10 μm.

Etymology. Referring to its occurrence on Sambucus.

Holotype. ENGLAND, Yorkshire, Worksop, Rotherham, Anston, Anstonstones Wood, on partly decorticated branches of Sambucus nigra, 16 May 2011, T. Læssøe et al. (WU 33556, culture RS = CBS 131707).

Pseudostromata erumpent from wood, eutypoid, very variable in shape and size, roundish to longish, extending from a few mm to several cm, covered by a grey to black, 20–80 μm thick ectostromatic zone, containing indistinct to white prosenchymatous entostroma and one to numerous perithecia; on bark only visible as minute circular ostiolar discs 50–180 μm diam, with umbilicate perforation, sometimes surrounded by some clypeate ectostroma. Ascomata immersed in wood, depressed globose, 0.4–0.8 mm diam, 0.25–0.5 mm high; peridium consisting of a dark brown, 15–40 μm thick pseudoparenchymatous outer layer and a narrow hyaline to yellowish hyphal inner layer. Paraphyses apically free, simple, 3–4 μm wide, attenuated to pointed at the apices. Asci (166–)175–228(–241) × (11.5–) 12.3–16.8(–21.5) μm (n = 20), cylindrical, with short stipe, thick-walled, appearing bitunicate and sometimes fissitunicate, containing (4–)8 uniseriate ascospores, apex thickened, with a small but distinct ocular chamber and a minute thin amyloid subapical ring (3.2–)3.3–4.1(–4.3) μm wide, (0.4–)0.5–0.9 (–1.2) μm high (n = 24), turning pinkish in Congo Red and blue in Waterman black ink. Ascospores (21–)24.5–31.3(–35.7) × (8.3–)9.0–11.2(–13.5) μm, l/w (2.1–)2.4–3.1(–4) (n = 117), oblong to ellipsoid, straight, rarely curved, light to yellow- or medium brown, with a central scarcely constricted euseptum and 2–4(–6) distosepta, i.e. 3–5(–7)-distoseptate, thick-walled, multiguttulate, when young with a thick sheath or small hyaline terminal caps visible in water and India ink but not stained by Waterman black ink nor aqueous nigrosin; sides sometimes pinched; occasionally with an oblique longitudinal septum; turning olive, sometimes with yellow contents in Lugol.

Culture characteristics — Growth on CMD and MEA slow, colony radius 27–37 mm after 1 mo at 20 °C; colony white, turning yellow-brown to black from the centre, odour lacking.

Ecology — On mostly decorticated branches of Sambucus nigra lying on the ground, sometimes submerged in aquatic habitats.

Distribution — Europe (England, France, Germany).

Additional material examined. ENGLAND, Buckinghamshire, High Wycombe, Hughenden Manor grounds (National Trust), grid reference SU 860 955, on Sambucus nigra, 30 Mar. 2013, P. Thompson (WU 33557, culture RS5); East Cornwall, between Liskeard and Callington, Gang, UK grid reference SX307681 (N50°29’17" W4°23’17"), on Sambucus nigra, 9 Mar. 2013, K. PrestonMafham, comm. P. Cannon. – FRANCE, Ariège, Castelnau-Durban, Artillac stream, down-stream from the marble quarry, elev. 410 m, on a submerged decorticated branch of Sambucus nigra, 17 Aug. 2015, J. Fournier JF15060; Rimont, La Maille, elev. 550 m, on a submerged decorticated branch of Sambucus nigra, soc. Melanomma fuscidulum, Aquaticola spp., 2 Apr. 2013, J. Fournier JF13064 (WU 33558, culture RS6); same area, La Maille brook, elev. 550 m, submerged decorticated branch of Sambucus nigra, 2 Oct. 2013, J. Fournier JF13177. – GERMANY, Mecklenburg-Vorpommern, Westmecklenburg, S from Rehna, forest ‘Benziner Tannen’, on Sambucus nigra, 15 July 2012, T. Richter, comm. H.-O. Baral and G. Friebes (WU 33559).

Notes — Unlike Lepteutypa fuckelii, where perithecia are immersed in bark, L. sambuci forms its ascomata immersed in the wood, resembling in this respect L. hederae, which differs by consistently 3-septate ascospores lacking a median euseptum and by inamyloid ascus apices.

Lepteutypa uniseptata (K.M. Tsui et al.) Jaklitsch & Voglmayr, comb. nov. — MycoBank MB814830

Basionym. Clypeosphaeria uniseptata K.M. Tsui et al., Mycologia 93: 1004. 2001.

Notes — All morphological criteria of L. uniseptata used in the protologue to justify its affiliation to the genus Clypeosphaeria such as the presence of a clypeus, long cylindrical asci with subapical, amyloid rings and uniseriate, ellipsoidal, brown ascospores fit to Lepteutypa, which was not considered. Particularly differences from the generic type Clypeosphaeria mamillana, such as a discoid apical ring in C. uniseptata (vs wedge-shaped in C. mamillana), thick-walled ascospores in C. uniseptata, but also the absence of an asexual morph are all typical of Lepteutypa. The shape or structure of the apical discharge apparatus in the ascus appears to be genus-specific in certain groups of the Xylariales (see e.g. Jaklitsch et al. 2014). Thus, the only difference to other species of Lepteutypa is ascospore septation, which can be regarded as negligible when considering the variable septation among other species of Lepteutypa and the clear phylogenetic affiliation of L. uniseptata to L. fuckelii.

Beltraniaceae Nann. 1934

Beltraniella carolinensis (M.E. Barr & Hodges) Jaklitsch & Voglmayr, comb. nov. — MycoBank MB814945

Basionym. Pseudomassaria carolinensis M.E. Barr & Hodges, Mycologia 63: 562. 1971.

Notes — Pseudomassaria carolinensis is phylogenetically distinct from P. chondrospora but contained within the highly supported Beltraniella clade (Fig. 1, as Beltraniella carolinensis). Hodges & Barr (1971) obtained a Beltraniella asexual morph in pure cultures of Pseudomassaria carolinensis, which they tentatively attributed to B. portoricensis (F. Stevens) Piroz. & S.D. Patil. However, they already raised serious doubts about conspecificity due to numerous differences, i.e. smooth setae (verrucose in B. portoricensis; see also key in Castañeda Ruiz et al. 1996); frequent production of ascomata in nature as well as pure cultures (unknown in B. portoricensis and any other Beltraniella species); high host specificity, being constantly associated with ascomata of Phyllachora perseae in circular necrotic spots on living or recently shed leaves of Persea borboni (B. portoricensis being a common leaf litter fungus with low host specificity, widespread in tropical to mild temperate areas); and culture characteristics distinctly different from isolates of B. portoricensis obtained from leaf litter. Only an LSU but no ITS sequence is deposited for P. carolinensis at GenBank, but at the NBRC (Biological Resource Center, NITE, Japan, http://www.nbrc.nite.go.jp/) an ITS sequence is available for the authentic (likely ex-type) strain IFO 9502 (= NBRC 9502). This ITS sequence differs from all available sequences of B. portoricensis deposited at GenBank (GU905993, KJ512150) and NBRC. Considering the molecular, morphological as well as ecological differences, Pseudomassaria carolinensis is considered a distinct species and here combined in Beltraniella.

Pseudomassariaceae Senan. & K.D. Hyde 2015

Pseudapiospora corni (Sowerby) Petr., Hedwigia 68: 233. 1928 — Fig. 5

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Fig. 5

Pseudapiospora corni. a, b. Ascomata in face view, showing the distinct clypeus; c. black ascomata exposed by epidermis cracks; d. perithecium wall; e–g. immature asci (e. in 3 % KOH + Lugol; f, g. in 3 % KOH); h. mature ascus; i, j. ascus apices in side (i) and face (j) view (in Lugol); k–r. ascospores (r. in Lugol) (a–f, h–r: WU 36852; g: WU 36853). — Scale bars: a = 1 mm; b, c = 0.2 mm; d–r = 10 μm.

Basionym. Sphaeria corni Sowerby, Col. Fig. Engl. Fung. Mushr. (London) 3, 25: pl. 370. 1802.

Pseudomassaria corni (Sowerby) Arx, Ber. Schweiz. Bot. Ges. 62: 349. 1952.

For additional synonyms see Barr (1964).

Typification. Neotype, here designated: AUSTRIA, Vienna, 3rd district, Botanical Garden of the University, in bark of Cornus alba cv. elegantissima, 21 Aug. 2015, H. Voglmayr (WU 36852; MBT 202954; ex-neotype culture PCO = CBS 140736).

Ascomata perithecial, scattered, immersed below rounded, distinct, greyish brown to blackish clypei in slightly elevated epidermis of the host, depressed globose, 170–250 μm wide, corresponding to the width of the clypeus, black. Peridium 10–20 μm wide at the base, thickened up to 50 μm toward the apex, of dark brown pseudoparenchymatous cells 5–22 μm diam. Hamathecium of apically free, sparsely branched paraphyses embedded in gel, distinct in water, indistinct in 3 % KOH, 1.5–6 μm wide. Asci in water (60–)65–88(–94) × (17–)18–21(–23) μm (n = 12), broadly fusoid, unitunicate, unstable when fresh, disintegrated by pressure, clavate, with 8 irregularly biseriate ascospores and short narrow stipe; apex rounded to attenuated, thickened to c. 1.5 μm, containing an amyloid ring 2.5–3.7 × c. 1 μm, indistinct in water, distinct in 3 % KOH. Ascospores (17.5–)20–23.5(–25.5) × (6.5–)7.5–9.0(–9.5) μm, l/w (2.1–)2.4–2.9(–3.3) (n = 72), broadly ellipsoid to clavate, with rounded ends, apiosporous with the septum at or above the lower third of the ascospore, small lower cell (7–)8–10(–11) μm (n = 25) long; straight or curved, thick-walled, smooth; contents of vital spores multiguttulate, commonly with a large and several smaller guttules per cell.

Colonies on MEA and CMD first hyaline to pure white, becoming dark olive grey with age, aerial mycelium cottony on MEA, scant on CMD; reverse first dull whitish, becoming black with age; growth radius at 22 °C after 6 wk to c. 60 mm on CMD, to c. 43 mm on MEA. Ascomata produced on MEA after c. 1 mo, (160–)190–400(–550) μm wide. Asci in 3 % KOH (83–)85–101(–108) × (14–)14.3–16.7(–17.5) μm (n = 7), oblong to fusoid. Ascospores in 3 % KOH (15.5–)19.3–24.5(–26) × (5.3–)6.0–7.2 μm, l/w (2.8–)3.1–3.6(–4) (n = 13), similar to, but slightly smaller than, those from ascomata on the natural substrate, with a distinct gelatinous sheath in the ascus. No asexual morph observed.

Habitat — In bark of Cornus alba and C. sericea.

Distribution — Europe, North America.

Additional material examined. AUSTRIA, Kärnten, Griffen, parking lot of Mochoritsch Griffen-Rast, in bark of Cornus alba, 3 Nov. 2015, H. Voglmayr & W. Jaklitsch (WU 36855); Oberösterreich, St. Willibald, communal tennis court, in bark of Cornus alba cv. elegantissima, 30 Aug. 2015, H. Voglmayr (WU 36854). – FRANCE, 21, Til-Châtel, Le Tertre, 19 Aug. 2015, A. Gardiennet AG15055 (WU 36853, culture PCO1).

Notes — Sowerby (1802) described and illustrated Sphaeria corni, the basionym, from Cornus suecica. No type material is extant in K (B. Auguirre-Hudson, pers. comm.), where the Sowerby collection is maintained after transfer from BM, and the original illustration is insufficient to serve as lectotype. No specimens from the type host are known to us, but in the lack of a type specimen we consider it desirable to propose a neotype from Cornus alba to maintain the species concept in its current use, in accordance with earlier detailed descriptions (e.g. Petrak 1925, Müller & Von Arx 1962, Barr 1964, Corlett 1978, 1981). All these descriptions were based on collections from Cornus alba or the closely related C. sericea.

Pseudapiospora corni typically occurs on Cornus alba on which it is common. We have not seen it on C. sanguinea, and we are not aware of any confirmed records from that host. Senanayake et al. (2015) alleged to treat this fungus under the name Pseudomassaria corni. Based on a specimen collected by E. Camporesi from Cornus sanguinea in Italy, they ignored all earlier detailed descriptions (see above), and made a description and illustration of an immature ascomycete with unicellular ascospores. Apparently this specimen also contains Pseudomassariella vexata, because the ITS sequence (KR092791) obtained from this specimen and deposited by Senanayake et al. (2015) in GenBank represents Pseudomassariella vexata. Apart from few obvious sequencing errors at the very beginning and the end, the sequence is identical with the P. vexata ex-epitype sequence JF440977 published by Jaklitsch & Voglmayr (2012). At last they defined this specimen even as ‘reference specimen of Pseudomassaria corni’, which, however, has no formal status as it is not governed by the ICN.

Pseudomassariella vexata (Sacc.) Petr., Sydowia 9: 603. 1955

Basionym. Didymella vexata Sacc., Michelia 2 (no. 6): 58. 1880.

Leiosphaerella vexata (Sacc.) E. Müll., in Müller & Von Arx, Beitr. Kryptogamenfl. Schweiz 11 (no. 2): 674. 1962.

Typification. Holotype of Didymella vexata: FRANCE, Rouen, Quevilly, on corticated twigs of Cornus sanguinea, J.B.P. Letendre (PAD). Epitype, here designated: AUSTRIA, Vienna, 3rd district, Botanical Garden of the University, on corticated twigs of Cornus sanguinea, 9 Jan. 2011, H. Voglmayr (WU 31333, ex-epitype culture CBS 129022 = LVE; MBT203100; ITS-LSU sequence JF440977).

Notes — When Fuckel (1871) combined Sphaeria oblitescens in Didymosphaeria, he gave a description of a specimen of his own, which corresponds to P. vexata. However, the type of Sphaeria oblitescens represents a true Didymosphaeria (Scheinpflug 1958). Fuckel (1871) noticed differences in the description and illustration from the original fungus, but ascribed this to the “younger state of the fungus in his specimen”. Saccardo (1880) received French material from J.B.P. Letendre, noticed that it conforms to Fuckel’s fungus and described it as Didymella vexata. In the Didymella folder in PAD there are two specimens labelled D. vexata, one collected by Krieger, and a second specimen without any collection data on the specimen which we identify as the holotype. The latter contains a short hand-written Latin description by Saccardo, a drawing of perithecium, asci and ascospores labelled with size ranges identical to those given in the original description, and several small corticated twig fragments. However, the material is scant and not sent out on loan, therefore we epitypify the name with a recently collected specimen which was illustrated in Jaklitsch & Voglmayr (2012) and for which a culture and sequences are available. See Jaklitsch & Voglmayr (2012) for a detailed description and illustration. Placement of Pseudomassariella in Pseudomassariaceae receives no bootstrap support (Fig. 1), familial affiliation is therefore provisional.

Requienellaceae Boise 1986, emend.

Type genus. Requienella Fabre. Other genus in the family: Acrocordiella O.E. Erikss.

Ascomata immersed or erumpent, subglobose; ostiolar necks inconspicuous or massive and strongly erumpent, black; extra-ascomatal tissue present. Hamathecium comprising two types of apically free paraphyses differing in length and width. Asci bitunicate, fissitunicate, cylindrical, subfusiform to narrowly clavate, with thick-walled apex, wide ocular chamber comprising a slightly refractive, inversely funnel-shaped dome turning slightly reddish in Congo Red, containing 8 uni- to biseriately arranged ascospores. Ascospores ellipsoid to oblong, olivaceous or brown, sometimes with lighter ends, with several transverse distosepta and large lumina.

Acrocordiella O.E. Erikss., Mycotaxon 15: 189. 1982

Type species. Acrocordiella occulta (Romell) O.E. Erikss.

Acrocordiella was described by Eriksson (1982) for the single species Massaria occulta, compared by him with Acrocordia and Pyrenula and assigned to the Pyrenulaceae. Later Boise (1986) and Barr (1990b) treated the genus as a synonym of Requienella. Barr (pers. comm., Jan. 2001) concluded that the genus is “quite separate from Requienella”, upon examination of a specimen (W.J. 1537) collected by the senior author in Austria. In order to test this hypothesis, fresh material of this fungus and of Requienella species were recollected, cultured and sequenced. Phylogenetic analyses as well as morphological re-investigations confirmed Acrocordiella to represent a distinct genus closely related to Requienella (see below).

Acrocordiella occulta (Romell) O.E. Erikss., Mycotaxon 15: 189. 1982 — Fig. 6

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Fig. 6

Acrocordiella occulta. a, b. Ostioles and white spots on the bark surface; c. ascoma in vertical section; d. ascomata in transverse section; e. ascomatal wall in section; f, g. asci; h. ascus apex in aqueous Cotton Blue; i. paraphysis between young asci; j. elongated paraphysis in aqueous Cotton Blue; k–s. ascospores (k, n. in 3 % KOH; o, p. dead, in water; q. in Lugol; r. in Cotton Blue/lactic acid; s. in Congo Red) (a, k: GZU; b–j, l–s: WU 33551). — Scale bars: a, d = 0.5 mm; b = 0.8 mm; c = 150 μm; e, i, q = 10 μm; f, g = 15 μm; h, j–p, r, s = 7 μm.

Basionym. Massaria occulta Romell, Hedwigia 24: 262. 1885.

Hymenopleella occulta (Romell) Shoemaker & Leclair, Canad. J. Bot. 53: 1587. 1975.

Pseudovalsa occulta (Romell) Berl., Icon. Fungorum (Abellini) 1, 1: t. 34, f. 3. 1890.

Lectotype, here designated: SWEDEN, Närke, Kumla, on twigs of Ribes grossularia, 1885, L. Romell, in Winter, Fungi Eur. Extraeur. Exs. 3354 (S, MBT202955). Epitype, here designated: AUSTRIA, Osttirol, Virgental, Prägraten, Umbalfälle, elev. 1420 m, on dead twigs of Ribes petraeum attached to the shrubs, 17 June 2015, H. Voglmayr & W. Jaklitsch (WU 33551; MBT202956; ex-epitype culture RS9 = CBS 140500).

Ascomata virtually invisible on the unchanged or sometimes slightly lifted bark, but on the surface often indicated by small white spots 120–300 μm diam around ostiolar openings or independent from them; sometimes, particularly with overmature ascomata, by circular black, non-projecting 50–125 μm wide ostiolar openings with a sunken minute circular perforation. Ascomata immersed in bark, singly or in small groups, within the bark often surrounded by white mycelium of hyaline 2–4 μm wide hyphae, depressed globose to ellipsoid, black, 400–800 μm diam, c. 300–500 μm high. Ostioles containing terminally 1–2.5 μm wide periphyses. Peridium 20–40 μm thick, comprising a textura angularis of small thick-walled angular cells (2.5–)3.0–7.5(–12.5) μm (n = 32) diam. Hamathecium of apically free, scarcely branched, 2–5 μm wide regular paraphyses, sometimes with basal cells inflated to 7 μm, and numerous, variously curved, 1–2.5(–3) μm wide elongated paraphyses in a gel matrix. Asci emerging from the base and sides of the ascoma nearly up to the ostiole, (150–)160–183(–191) × (11.5–)12–16.5(–21.7) μm (n = 23), cylindrical, attenuated toward the apex, containing 8 (obliquely) uniseriate ascospores, not obviously fissitunicate, thin-walled; apex attenuated and distinctly thickened, containing a slightly refractive, inversely funnel-shaped canal 4–5.5 μm high, 3–5.7 μm diam, extending up to 11 μm in 3 % KOH, staining blue in aqueous Cotton Blue and faintly reddish in Congo Red; base simple or knob-like. Ascospores (16.5–)20–24.3(–27.7) × (6.8–)8.3–10(–11) μm, l/w (1.8–)2.2–2.7(–3.2) (n = 70), ellipsoid, ends narrowly rounded to nearly acute, straight to slightly curved, olivaceous, greyish olive when young, dark brown when dead, 3-distoseptate, lumina rhomboid to square-shaped, central ones slightly larger than terminal ones, multiguttulate, sides often slightly pinched, smooth, without sheath; in KOH more angular and eventually terminal distosepta disappearing; darkening in Lugol; turning pale brown in Cotton Blue/lactic acid.

Culture characteristics — Growth slow; decreased temperates (15–22 °C) required; colonies white. No asexual morph observed.

Habitat — In bark of Ribes spp.

Distribution — Central, Eastern and Northern Europe (Austria, Russia, Sweden).

Additional material examined. AUSTRIA, Osttirol, Virgental, Prägraten, Umbalfälle, elev. 1450 m, on dead twigs of Ribes petraeum, 17 June 2015, W. Jaklitsch & H. Voglmayr (WU 33552, culture RS10 = CBS 140501); ibid., 28 Aug. 2000, W. Jaklitsch W.J. 1537 (WU 33550). – SWEDEN, Närke, Kumla, on Ribes grossularia; 29 June 1889, L. Romell, Fungi exs. praes. scand. 73 (GZU).

Notes — As the epithet implies, A. occulta is an inconspicuous fungus, well hidden in the bark. This character, a simpler ascus apex, a different ascospore anatomy, different ecology and the results of phylogenetic analyses clearly demonstrate that Acrocordiella is not congeneric with Requienella. Previously only known from Ribes uva-crispa (= R. grossularia) in Scandinavia and from Ribes nigrum in Russia, it was also collected in Austria in 2000 (Hausknecht et al. 2003), but not cultured at that time. Cultures were prepared from specimens collected recently in the same area.

Requienella Fabre, Ann. Sci. Nat., Bot., sér. 6, 15: 55. 1883

= Trematomyces Schrantz, Bull. Soc. Mycol. France 76: 324. 1961.

Type species. Requienella seminuda (Pers.) Boise.

Boise (1986) described the family Requienellaceae for Requienella Fabre in the Dothideomycetes (as Loculoascomycetes). She lectotypified Requienella with R. olearum, determined its earliest epithet as Sphaeria seminuda Pers. 1801 and stated that the three other species recognised by Fabre (1883) in Requienella belonged to other genera. The family Requienellaceae was not accepted by Eriksson (in Eriksson & Hawksworth 1986) and he maintained Requienella in the Pyrenulaceae. He also supposed that the ‘non-branched pseudoparaphyses’ are true paraphyses, as in Pyrenula. Barr (1990b) listed the Requienellaceae as a family of her order Melanommatales. Aptroot (1991) accepted Barr’s system, treated the Pyrenulales as synonym of the Melanommatales and added five genera of lichenised fungi to the non-lichenised Requienella in the Requienellaceae. Harris (1995) considered to accept the Requienellaceae but being only restricted to the single genus Requienella. Kirk et al. (2008) listed the Requienellaceae as a family of the Pyrenulales. Here we show that Requienella surprisingly belongs to the Xylariales, despite its fissitunicate asci.

Requienella seminuda (Pers.) Boise, Mycologia 78: 38. 1986 — Fig. 7

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Fig. 7

Requienella seminuda. a, g. Ascomata in face view; b. ascomata in vertical section; c, d. asci (note apically free paraphyses in d); e. apically free paraphysis; f. free apex of elongated paraphysis (in Congo Red); h. ascus apex; i–k. ascospores (a, c, f, h, j: WU 33569; b, i: WU 33567; d, e, k: WU 33568; g: lectotype L0113128). — Scale bars: a, b, g = 0.2 mm; c, d = 15 μm; e, h–k = 7 μm; f = 10 μm.

Basionym. Sphaeria seminuda Pers., Syn. Meth. Fung. (Göttingen) 1: 70. 1801.

= Sphaeria olearum Castagne, Cat. Pl. Marseille: 116. 1845 [non Sphaeria olearum De Not., 1845, nom. illegit., art. 53.1].

≡ Caryospora olearum (Castagne) Sacc., Syll. Fung. (Abellini) 2: 118. 1883.

≡ Requienella olearum (Castagne) Fabre, Ann. Sci. Nat., Bot., sér. 6, 15: 1883.

Trematosphaeria olearum (Castagne) Sacc., Syll. Fung. (Abellini) 2: 118. 1883.

Trematomyces olearum (Castagne) Schrantz, Bull. Trimestriel Soc. Mycol. France 76: 323. 1961.

Typification. Lectotype of Sphaeria seminuda designated by Boise (1986):?FRANCE, Herb. Persoon L0113128 (= 910269-31, L!). Epitype, here designated: GREECE, Crete, Rethymno, Armeni, on bark of Olea europaea, 9 June 2015, H. Voglmayr & W. Jaklitsch (WU 33568; MBT 202957; ex-epitype culture RS12 = CBS 140502).

Ascomata solitary or aggregated in groups of 2–5, perithecioid, conical with rounded base or more or less globose with a prominent apex, immersed, large, (0.35–)0.45–0.9(–1.1) mm diam, 0.6–1.2 mm high, often height exceeding the diam, often surrounded by whitish, greyish to black matter. Ostiolar neck erumpent, massive, papillate to conical, shiny black, apex typically 25–160 μm wide, round, blunt to pointed, sometimes flattened and 90–300 μm diam. Peridium 15–40 μm wide at the base, thickened to 160 μm in upper regions, dark brown, consisting of minute thick-walled angular cells. Hamathecium consisting of 2–5 μm wide, apically free paraphyses similarly long as the asci, and numerous long, not or scarcely branched, 1–2.5 wide, apically free elongated paraphyses (‘pseudo-trabeculae’) nearly reaching the ostiolum, and periphyses of similar width in the ostiolum, all immersed in a gel matrix. Asci (148–)158–178(–182) × (21.7–)23.5–29(–32.5) μm (n = 13), bitunicate, fissitunicate, oblong to subfusiform, with thick-walled apex, wide ocular chamber comprising a slightly refractive, inversely funnel-shaped dome 5.5–7 μm high, 6.5–7.7 μm wide at the base, turning slightly reddish in Congo Red, demarcated by a basal plate, with short stipe and simple to knob-shaped base, containing 8 uni-, in the middle often biseriately arranged ascospores. Ascospores (25.3–)28.3–32.7(–37) × (9.8–)10.8–12.7(–13.3) μm, l/w (2.2–)2.3–2.9(–3.7) (n = 50), first hyaline, 1-celled, inequilateral, developing 1–3 septa and turning brown with slightly paler end cells, becoming (3–)5(–7)-distoseptate, ellipsoid, with the width lumina exceeding their length, turning darker olivaceous in KOH and lumina becoming smaller and more angular.

Culture characteristics — Growth of cultures extremely slow (few mm per month), only somewhat enhanced by overlay of sterile water; colonies white. No asexual morph observed.

Habitat — In dead bark, less commonly wood of living trunks of old Olea europaea trees.

Distribution — Southern and Western Europe (France, Greece, Italy).

Additional material examined (all from bark of Olea europaea). GREECE, Crete, Chania, Platanias, Pananiana, 4 June 2015, H. Voglmayr & W. Jaklitsch (WU 33566); Rethymno, Ambelaki, 9 June 2015, W. Jaklitsch & H. Voglmayr (WU 33567, culture RS11); Vizari, 9 June 2015, W. Jaklitsch & H. Voglmayr (WU 33569, culture RS13).

Notes — Requienella seminuda is a peculiar fungus. Its distinctly fissitunicate asci have prevented a placement outside groups of ascomycetes with bitunicate asci. The hamathecium has never been studied in detail resulting in controversial reports: Barr (1990b) found narrow, scarcely branched trabeculae, whereas Eriksson (in Eriksson & Hawksworth 1986) anticipated that the ‘non-branched pseudoparaphyses’ are probably true paraphyses. Peculiar is the fact that three kinds of threads are present, a mass of numerous long and extremely narrow hyphae with free ends extending to the upper part of the ascoma (‘pseudo-trabeculae’), wider ‘normal’ paraphyses between asci, best seen among immature asci, and periphyses in the ostiole. It is difficult to determine the nature of the entire hamathecium. As a matter of fact, all threads appear to have free ends, which suggests that some paraphyses are strongly elongated and taper upwards forming long straight ‘pseudo-trabeculae’ at upper levels of the ascoma. In any case, the presence of true paraphyses is in line with other Sordariomycetes, as are the sequences of the phylogenetic markers used here. According to Fabre (1883) and Saccardo (1883) ascospores of R. seminuda are 5- or more-septate. We found only rarely a number of septa deviating from five.

Nomenclatural background: Sphaeria olearum was based on a specimen collected by Desmazières. Boise (1986), upon examination of an ?isotype of this material from NY and of Persoon’s type material of Sphaeria seminuda from Leiden, determined that they represent the same species, that S. seminuda is older than S. olearum and chose Persoon’s material as lectotype. She also illustrated regularly 5-distoseptate ascospores for Persoon’s material L0113128 (= 910269-31, L!). Later Aptroot (1991) examined the original type material of Sphaeria olearum (France, Bouches-du-Rhône, Desmazières 1767) present in G and confirmed conspecificity of the two taxa. We examined and illustrate ascomata of the lectotype of R. seminuda. As Boise (1986) illustrated 5-distoseptate ascospores for the lectotype, we conclude that Persoon’s material, like Desmazières’s original material, originates from Olea europaea, most probably from France. We epitypify S. seminuda with material from Olea, due to an earlier mix-up of R. seminuda by several authors with Acrocordiella occulta and Requienella fraxini described below.

Requienella fraxini Jaklitsch & Voglmayr, sp. nov. — MycoBank MB814828; Fig. 8

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Fig. 8

Requienella fraxini. a–d. Ascomata in face and lateral views; e. ascoma in vertical section (note wall thickening in ostiolar region); f. hamathecium in the upper ascomatal region; g. young asci with paraphyses; h. free apical ends of paraphyses; i, v. asci (i in Lugol; note fissitunicate dehiscence in v); j–l. ascus apices (k. in Lugol + Congo Red; l. in Lugol); m–u. ascospores (m, n. immature; o–q. vital) (a, b: WU 33560; c, d: WU 33564; e–i, k, n, s–v: WU 33561; j, l: WU31483; m, o–q: WU 33565; r: WU31482). — Scale bars: a–d = 0.5 mm; e = 0.3 mm; f, i, v = 25 μm; g = 50 μm; h, j, l–t = 10 μm; k, u = 15 μm.

Etymology. Referring to its occurrence on Fraxinus.

Holotype. NORWAY, Akershus, Bærum kommune, Jonsrudtjern, UTM 32: 0580011/6649407, at the steep slope of the small lake, soc. Navicella pileata, 18 Sept. 2012, B. Norden (WU 33561, ex-type culture RS3 = CBS 140475).

Ascomata perithecioid, immersed, with the ostiolum, often including an upper part of the ascoma, erumpent to 0.2–0.9 mm above the host surface, solitary or aggregated in small numbers, 0.5–1.1 mm high, 0.45–1 mm diam, conical with rounded base or more or less globose with a prominent, conical, more or less acute, shiny black, apically 26–160 μm wide papilla, circular in transverse section, black; often surrounded by white amorphous tissue containing some hyaline 2–6 μm wide hyphae; tissue sometimes turning black by ejected ascospores. Peridium 15–30 μm wide at the base, thickened to 150 μm and hard in upper regions, dark brown, consisting of small pseudoparenchymatous cells. Hamathecium complex, consisting of 2–5 μm wide, apically free paraphyses containing oil drops when vital, and similarly long as the asci, superposed by masses of long, sparcely branched, 1–2.5 wide, apically free ‘pseudo-trabeculae’ nearly reaching the ostiolum and variously curved periphyses of same width in the ostiolum, all immersed in a gel matrix. Asci 153–206 × 20–30(–33) μm (n = 10), bitunicate, fissitunicate, oblong to narrowly clavate, with thick-walled apex, wide ocular chamber comprising a slightly refractive, inversely funnel-shaped dome 9–11 μm long, 6.5–9.5 μm wide at the base, turning reddish in Congo Red, demarcated by a basal plate, with short simple stipe, containing 8 uni- to biseriately arranged ascospores. Ascospores (23.3–)26.7–31.5(–36) × (8.0–)9.5–12(–14.5) μm, l/w (2.1–)2.4–3.1(–3.9) (n = 100), ellipsoid, oblong to fusiform, first hyaline, 1-celled, with narrow sheath, becoming septate and yellow, finally brown with lighter ends, 3(–5)-distoseptate, with large lumina and faintly punctate perispore.

Culture characteristics — Growth of cultures extremely slow, only somewhat enhanced by overlay of sterile water; colonies white. No asexual morph observed.

Habitat — In bark of living trunks of old trees of Fraxinus excelsior.

Distribution — Northern and Western Europe (France, Norway, Sweden), probably North America.

Additional material examined (all from bark of Fraxinus excelsior). FRANCE, Poitou-Charentes, Le Vanneau, Marais Poitevin, 30 Oct. 2013, A. Gardiennet AG13205 (WU 33564 = RS7; culture lost). – NORWAY, Akershus, Bærum kommune, Jonsrudtjern, UTM 32: 0580020/6649407, 18 Sept. 2012, B. Norden (WU 33560, culture RS2); Akershus, Rogaland kommune, Strand, Rag, UTM 32: 26104/55362, 4 Oct. 2012, B. Norden & J.B. Jordal (WU 33563 = RS4); Bærum kommune, Tanumbråtan north, UTM 32: 0582366/6640274, 20 Sept. 2012, B. Norden (WU 33562); Vest-Agder, Kristiansand kommune, Nedre Timenes, 5 Oct. 2014, H. Voglmayr & W. Jaklitsch (WU 33565, culture RS8 = CBS 140476). – SWEDEN, Västra Götaland, Bohuslän, Uddevalla, Ljungskile, 1 Apr. 2001, N. Vassbosjön (WU 31582); Blekinge, Angölsmåla, Olofström, Kyrkhult, 27 Apr. 2002, B. Norden (WU 31583).

Notes — Prior to the recent recollection of Requienella seminuda on Olea europaea we identified R. fraxini as R. seminuda, following Boise (1986). However, molecular data clearly separate these two species. Moreover, the description of R. seminuda by Boise (1986), Aptroot (1991) and Barr (1990b) as having regularly 3–6-, or 3–7-septate ascospores suggested that the different septations occur at comparable frequencies. This is, however, not the case, as ascospores of R. seminuda are regularly 5-septate and those of R. fraxini 3-septate. In each species only a very small fraction may have deviant numbers of septa. This is also supported by the description of ascospores for R. olearum as 5- or more septate by all authors dealing with the taxon in the 19th century. Distinction criteria other than ascospore septation are different hosts (Olea vs Fraxinus) and the geographical distribution, R. fraxini in northern humid regions, R. seminuda in southern mediterranean regions of Europe, with southern France as an overlapping zone. Requienella fraxini appears to be confined to old trees in areas of high humidity and is therefore much rarer than its host. It may be in decline and endangered due to ash dieback. Nordén & Jordal (2014) gave an account of R. fraxini under the name R. seminuda, i.e. before we collected R. seminuda on Olea.

Sporocadaceae Corda (as ‘Sporocadeae’), Icon. Fungorum (Prague) 5: 34. 1842

Type genus. Sporocadus Corda, Icon. Fungorum (Prague) 3: 23. 1839.

(synonymous with Seimatosporium Corda, in Sturm, Deutschl. Fl., Abt. 3, Pilze Deutschl. 3: 79. 1833.)

Hughes (1958) lectotypified Sporocadus with S. lichenicola, which is currently known as the plurivorous species Seimatosporium lichenicola, the asexual morph of Discostroma corticola (Shoemaker & Müller 1964), currently being treated as a synonym of D. fuscellum.

Hymenopleella Munk, Dansk Bot. Ark. 15: 89. 1953

Type species. Hymenopleella hippophaëicola Jaklitsch & Voglmayr.

Ascomata perithecial, immersed in bark, depressed globose. Ostioles periphysate. Peridium pseudoparenchymatous, 2-layered. Hamathecium of apically free paraphyses. Asci cylindrical, with 8 uni- to partially biseriate ascospores, containing a flat, V-shaped to sinuous, amyloid apical ring. Ascospores oblong, ellipsoid or fusoid, symmetric, with several eusepta, light yellow- to reddish brown wall and septa more strongly pigmented than contents, smooth, partly turning dull green in Lugol, lacking a sheath. Asexual morph monochaetia-like.

Hymenopleella hippophaëicola Jaklitsch & Voglmayr, nom. nov. — MycoBank MB814829; Fig. 9

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Fig. 9

Hymenopleella hippophaëicola. a. Ostioles and bumps on the bark surface; b, c. ascus rings in Lugol; d–f. asci (d, e. in 3 % KOH); g–i. ascospores (g. in 3 % KOH; i. in Lugol) (a, b, d, e, g: B700021868; c, f, h, i: WU 32027). — Scale bars: a = 0.2 mm; b, c = 5 μm; d–f = 15 μm; g–i = 10 μm.

Etymology. Referring to its occurrence on Hippophaë.

Replaced synonym. Sphaeria hippophaës Sollm., Bot. Zeitung 20: 379. 1862, non Hymenopleella hippophaës (Fabre) Rulamort, Bull. Soc. Bot. Centre-Ouest, Nouv. sér. 17: 192. 1986.

Leptosphaeria hippophaës (Sollm.) Sacc., Syll. Fung. (Abellini) 22: 222. 1913.

Lepteutypa hippophaës (Sollm.) Arx, Gen. Fungi Sporul. Cult. (Lehr): 118. 1970.

Massaria hippophaës (Sollm.) Jacz., Bull. Herb. Boissier 2: 684. 1894.

Hymenopleella sollmannii Shoemaker & E. Müll., Canad. J. Bot. 43: 1457. nom. inval. Art. 52.1

= Melanomma hippophaës Fabre, Ann. Sci. Nat., Bot., sér. 6, 9: 92. 1878.

Hymenopleella hippophaës (Fabre) Munk, Dansk Bot. Ark. 15: 90. 1953, nom. inval. Art. 41.5

Hymenopleella hippophaës (Fabre) Rulamort, Bull. Soc. Bot. Centre-Ouest, Nouv. sér. 17: 192. 1986.

Typification. No holotype seems to be extant, but there is an authentic specimen present in B, which was collected by the author of the replaced synonym, August Sollmann, two years after his publication. We designate this specimen as neotype of Sphaeria hippophaës: GERMANY, Coburg, on twigs of Hippophaë rhamnoides, 1864, A. Sollmann (B700021868; MBT203103). Epitype, here designated: AUSTRIA, Niederösterreich, Gerasdorf, Marchfeldkanalweg, on twigs of Hippophaë rhamnoides, 12 Aug. 2012, W. Jaklitsch (WU 32027; MBT202958; ex-epitype culture LH = CBS 140410).

Ascomata perithecial, immersed in bark, scattered or in small groups, 0.2–0.4(–0.5) mm diam, lifting the bark only slightly producing inconspicuous irregular bumps with small black ostiolar dots 30–80 μm diam. Ostioles 25–50 μm wide inside, central, short-papillate, periphysate; peridium 20–40 μm thick, pseudoparenchymatous, of a thin hyaline inner layer and a dense small-celled dull orange-brown outer textura angularis, sometimes surrounded by some whitish, buff to brown hyphal tissue. Paraphyses uncommon, simple, rarely branched, 3–5 μm wide. Asci (90–)97–126(–134) × (12–)12.5–16(–17) μm (n = 14), cylindrical, with 8 (partly obliquely) uniseriate, rarely more or less biseriate ascospores, with a short stipe; apex thickened to 2 μm (to 3.5 μm when young), containing a flat, V-shaped to sinuous, amyloid apical ring (4.0–)4.5–5.3(–5.5) × (0.7–)0.8–1.2(–1.5) μm (n = 17). Ascospores (15.2–)17.5–22.5(–26.5) × (7.2–)8.0–9.0(–9.7) μm, l/w (1.8–)2.1–2.7(–3.3) (n = 100), oblong, ellipsoid or fusoid, symmetric, ends rounded or attenuated, with 3 non-constricted eusepta, straight or slightly curved, light yellow- to reddish brown, darker brown in KOH, wall and septa more strongly pigmented than contents, smooth, partly turning dull green in Lugol, lacking a sheath.

Culture characteristics — Growth better on MEA than on CMD and better at 15 °C than at room temperature; colony first whitish, turning greyish brown to olive. No asexual morph observed here, but reported by Shoemaker & Müller (1965; under Hymenopleella sollmannii) from malt agar: Conidia formed in spherical cavities 140–300 μm diam, surrounded by a 15–50 μm thick wall of reddish brown hyphae. Conidiophores hyaline, simple, 10–30 × 2–3 μm, 1–2-septate, with fine annellations. Conidia fusoid, 20–27 × 6–9 μm, straight or rarely curved, mostly 4-septate, with medium yellow-brown central cells and colourless end cells, apical cell with an acuminate 3–6 μm long appendage sometimes separated by an additional septum, basal cell narrowed to 3 μm wide truncate base usually bearing a central, 2–4 μm long appendage.

Habitat — In bark of Hippophaë rhamnoides.

Distribution — Europe.

Additional material examined. AUSTRIA, Vienna, 21st district, Donauinsel, grid square 7764/3, on twigs of Hippophaë rhamnoides, 20 June 1999, W. Jaklitsch W.J. 1326 (WU 32027).

Notes — A detailed description of Hymenopleella hippophaëicola was given by Petrak (1947) under the name Leptosphaeria hippophaës. Ascospores mature asynchronously, starting from the ascus base. The taxonomic and nomenclatural history of this species is complicated. Munk (1953) based Hymenopleella on Melanomma hippophaës Fabre, which is younger than Sphaeria hippophaës Sollm. Winter (1887) recognised that these heterotypic names represent the same species, which was confirmed by Shoemaker & Müller (1965). Therefore Shoemaker & Müller (1965) introduced the new name Hymenopleella sollmanni as a replaced synonym for Sphaeria hippophaës Sollm., non Hymenopleella hippophaës (Fabre) Munk. However, they missed that, according to Art. 41.5 of the ICN, the combination Hymenopleella hippophaës (Fabre) Munk is invalid, as Munk (1953) did not give a reference to the place of valid publication of the basionym. Therefore, according to Art. 52.1 of the ICN, Hymenopleella sollmannii is a superfluous, illegitimate new name. However, since De Rulamort (1986) validated the combination Hymenopleella hippophaës based on Melanomma hippophaës Fabre, Sphaeria hippophaës Sollm. cannot be combined in Hymenopleella any more, and a new replacement name is necessary. Shoemaker & Müller (1965) accepted Hymenopleella as different from Lepteutypa due to the ascospores of Lepteutypa fuckelii, in being ‘narrower and thinner-walled, octagonal in section and having a sheath and a peculiar brown deposit in the middle line of each septum’. In our experience, a more striking difference is the euseptate nature of ascospores in H. hippophaëicola as opposed to distoseptate in L. fuckelii.

Seiridium Nees, Syst. Pilze (Würzburg): 22. 1816

Type species. Seiridium marginatum Nees.

Seiridium is characterised by acervular coelomycetes with versicolorous, 5-septate, appendaged conidia. Its type species Seiridium marginatum Nees 1816 (not Seiridium marginatum Schwein. 1832, a later homonym representing a rust fungus species of Phragmidium), was redescribed by Sutton (1980) and Shoemaker et al. (1966), who described its sexual morph as Blogiascospora marginata, based on co-occurrence. Here we confirm this relationship using DNA data.

Seiridium marginatum Nees, Syst. Pilze (Würzburg): 23. 1816— Fig. 10

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Fig. 10

Seiridium marginatum (WU 33575). a. Ascomata in bark fissure; b. ascoma in face view; c. conidioma in face view; d–f. asci (e. in Lugol; f. upper part with thickened apex); g, h. ascus apex in Lugol; i–k. ascospores (i. in 3 % KOH; k. in Lugol; note the pitted surface); l. conidiophores; m–q. conidia (o, p. detail in section and surface view). — Scale bars: a–c = 0.3 mm; d, e, l = 20 μm; f, h, i, m, n, q = 10 μm; g, j, k, o, p = 5 μm.

(non Seiridium marginatum Schwein., Trans. Amer. Philos. Soc., n.s. 4: 306. 1832 [1834].)

= Blogiascospora marginata (Fuckel) Shoemaker, E. Müll. & Morgan-Jones, Canad. J. Bot. 44: 248. 1966.

Massaria marginata Fuckel, Jahrb. Nassauischen Vereins Naturk. 27–28: 28. 1874.

Typification. Neotype of Seiridium marginatum, proposed by Shoemaker et al. (1966): SWITZERLAND, Jura, on twigs of Rosa, in spring, P. Morthier, in Fuckel, Fungi Rhen. Exs. 2136 (previously BM; now K(M) 200376). Epitype, here designated: FRANCE, Burgundy, Côte-d’Or, Til-Chatel, on twigs of Rosa canina; holomorph, 1 Apr. 2015, A. Gardiennet AG15018 (WU 33575; MBT202960; ex-epitype culture from ascospores: CBS 140403 = BLO; culture from conidia: SEI).

Sexual morph visible through bark fissures as pale brownish spots with a central black papilla or, after removal of upper bark layers, as flattened greyish to brownish pustules 0.2–0.6 mm diam. Ascomata perithecial, immersed in bark, depressed globose to pyriform, 0.3–0.6 × 0.2–0.5 mm, scattered or confluent in numbers of 2–3; peridium 20–30 μm thick, dark brown, pseudoparenchymatous. Ostioles central, slightly papillate, black, circular in section, periphysate. Paraphyses filiform, 1.5–2 μm wide at the free apex, widening to 4.5 μm downwards. Asci cylindrical, (145–)150–178(–190) × (9.0–)10.5–14.8(–16) μm (n = 10), containing 8 (obliquely) uniseriate ascospores, unitunicate, thin-walled, with a knob-like base and crozier and a slightly thickened apex containing an amyloid ring (3.2–)3.5–4.5(–4.8) μm wide, (0.6–)0.9–1.6(–1.8) μm high (n = 21). Ascospores (17.5–)21–25.7(–32.5) × (8.0–)8.5–10(–11.5) μm, l/w (2.1–)2.3–2.8(–3.7) (n = 56), cylindrical-oblong, with 3, rarely 5 thick eusepta often thicker than the wall, yellow- to dark brown, darkening in KOH and Lugol, multiguttulate when fresh and often with irregularly disposed minute light dots 0.5–1 μm diam. Conidiomata on the natural host acervular but often with a narrow opening, depressed globose, 0.3–0.6 mm diam, 0.2–0.4 mm high, immersed-erumpent, dehiscent by irregular crumbling, scattered or confluent, dark grey to black. Basal wall pseudoparenchymatous, of small dark brown cells. Conidiophores hyaline, cylindrical, more or less straight, sparsely and mostly asymmetrically branched near the base, up to c. 90 × 4 μm, branches up to 60 μm long. Conidia formed terminally, (35.2–)38.2–42(–45) × (8.2–)8.8–10.2(–11) μm, l/w (3.5–)3.9–4.6(–4.9) (n = 51), fusiform, straight, with 5 thick, scarcely constricted eusepta, versicolorous, of 4 brown cells (24.5–35) μm long, with striate surface and hyaline end cells, each with a hyaline unbranched filiform appendage to 52 μm long.

Habitat — In bark of Rosa canina.

Distribution — Europe (France, Germany, Switzerland).

Additional material examined. AUSTRIA, Niederösterreich, Hardegg, Maxplateau, on twigs of Rosa canina, 27 Apr. 2015, H. Voglmayr (WU 33576, culture SEI1 = CBS 140404).

Notes — A detailed description of the sexual morph was given by Shoemaker & Leclair (1975); see also Fuckel (1874) under Massaria marginata. Although Seiridium marginatum is morphologically well-defined, its host genus is extremely diverse and distributed all over the world. Therefore, it cannot be ruled out that morphologically similar but phylogenetically deviating species may appear in future. For this reason we fix the application of the name via epitypification.

Strickeria Körb., Parerga Lichenol. (Breslau) 5: 400. 1865

Type species. Strickeria kochii Körb.

Strickeria was originally described as a genus of lichenised fungi (Körber 1865). Since then this generic name was used by several authors for some Dothideomycetes with brown muriform ascospores, often with Teichospora in synonymy (Winter 1887) or Strickeria as a synonym of Teichospora. Barr (1990a, b), however, treated them as distinct genera of Dothideomycetes. Eriksson (in Eriksson & Hawksworth 1991), upon examination of original material of S. kochii, determined that the asci are unitunicate and therefore Strickeria was classified as Sordariomycetes incertae sedis (Lumbsch & Huhndorf 2010) or Ascomycetes (Kirk et al. 2008). We show here that Eriksson’s observation was correct and determine the phylogenetic relationship of S. kochii in Xylariales.

Strickeria kochii Körb., Parerga Lichenol. (Breslau) 5: 400. 1865— Fig. 11

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Fig. 11

Strickeria kochii. a–e. Ascomata (b, c. in side view); f, g. free apical ends of paraphyses in Congo Red; h, i. asci; j. ascus apex; k–n. ascospores; o–q. asexual morph in culture (CMD, 17–24 d, 22 °C); o. pycnidia; p, q. conidia; r–t. asexual morph from the natural substrate (r. pycnidial wall; s, t. conidiophores and conidia) (a, b, d: WU 33572; e, m, n: lectotype W 1915-11805; p, q: WU 33570; c, f–l, o, r–t: WU 33571). — Scale bars: a–e = 100 μm; f, j–n, p–t = 10 μm; g–i = 15 μm; o = 150 μm.

= Teichospora pezizoides Sacc. & Speg., Michelia 1(no. 3): 350 (1878).

= Hendersonia fusarioides Sacc., Michelia 1: 213 (1878).

Scolicosporium pauciseptatum Constant., Mycotaxon 41(2): 467 (1991).

Lectotype, here designated: AUSTRIA, Oberösterreich, Kremsmünster, on bark of Robinia pseudoacacia, 1862, Clar. Poetsch, in Koerber, Lich. Sel. Germ. 264 (W 1915-11805!; MBT202961). Epitype, here designated due to the earlier confusion with genera of Dothideomycetes: AUSTRIA, Vienna, 22nd district, Lobau, near Panozzalacke, on bark of Robinia pseudoacacia, 7 Mar. 2015, W. Jaklitsch (WU 33571; MBT202962; ex-epitype culture CBS 140411 = C143).

Ascomata superficial, scattered, (110–)170–260(–315) μm diam (n = 20), first nearly globose, soon collapsing, discoid, cupulate or turbinate with circular outline, broadly attached or downward slightly attenuated (broadly turbinate); surface black, warted, peridium thick and hard, dark brown, pseudoparenchymatous. Apex often with a short central, 45–80 μm wide papilla, roundish, angular to slightly elongate, ostiole periphysate. Hamathecium of simple, 1.5–4.5 μm wide, apically free paraphyses. Asci (106–)110–129(–132) × (11–)11.5–13(–13.5) μm (n = 10), cylindrical, with 8 uniseriate ascospores, short stipe and thickened apex without a distinct discharge apparatus, in KOH ascus wall thicker, ocular chamber larger, sometimes branched (thickened apex indented). Ascospores (12–)14.8–19(–24.2) × (6.0–)6.5–7.8(–8.5) μm, l/w (1.7–)2.1–2.7(–3.3) (n = 72), ellipsoid or broadly fusiform, with 3 transverse non-constricted septa and 1 longitudinal septum in mid cells, brown, with (sub-)hyaline end cells, smooth.

Asexual morph in nature: Ascomata associated with minute black pycnidia, 70–105 μm diam, globose, warted, with central pore. Pycnidial wall pseudoparenchymatous, of dark brown angular to globose cells 5–13 μm diam. Conidiophores 1–3-celled, hyaline, simple, to c. 40 μm long. No phialides present. Conidia formed in fascicles, (23.5–)32.5–48.3(–51) × (4.7–)5.0–6.0(–6.3) μm, l/w (4.4–)6.0–8.8(–9.6) (n = 21), falcate, first hyaline, 1-celled, finally brown, with 3–4 non-constricted septa, thin-walled, mid cells cylindrical, end cells lighter, acute and curved.

Asexual morph in culture: Colonies on CMD at 22 °C reaching after 1 mo a radius of up to c. 45 mm, hyaline to whitish, dense, slightly zonate, centre turning brown due to minute dark dots; the latter developing to pycnidia after c. 2 weeks. Pycnidia 70–175 μm diam, globose to cylindrical, dark brown, smooth, containing short hyaline cylindrical conidiophores producing strongly curved falcate conidia 35–49 × 4.7–6.0 μm of 2–5 cylindrical to nearly globose brown mid cells, a subhyaline, acute end cell and a subhyaline cylindrical base cell.

Habitat — On bark, particularly in recessed regions of thick bark of living trunks of Robinia pseudoacacia.

Distribution — Europe (Austria, Germany, Italy).

Additional material examined (all from bark of standing trees of Robinia pseudoacacia). AUSTRIA, Niederösterreich, Gumpoldskirchen, from Melkerhof upward, 1 Mar. 2015, W. Jaklitsch & H. Voglmayr (WU 33570, culture C138); Manhartsbrunn, 8 Mar. 2015, W. Jaklitsch & H. Voglmayr (WU 33572, culture C146); Mannersdorf im Leithagebirge, Schweingraben, 14 Mar. 2015, H. Voglmayr & I. Greilhuber (WU 33573, culture C149); Mödling, Eichkogel, 19 Apr. 2015, W. Jaklitsch & H. Voglmayr (WU 33574). – ITALY, Tuscany, Florence, Tavarnuzze, roadside, close to the motorway A1, 29 Oct. 2015, W. Jaklitsch (WU 36856).

Notes — Maturation of asci of Strickeria kochii in nature is asynchronous and due to the habitat-related exposition of the ascomata to drought many asci are not optimally developed or overmature in most specimens. Often they contain aberrant ascospores, with up to 8 septa and up to 33 × 7 μm.

Teichospora pezizoides and Hendersonia fusarioides, which were also described from Robinia, are evidently synonymous with Strickeria kochii; for a description and illustrations of H. fusarioides (as Scolicosporium pauciseptatum) see Constaninescu (1991).

Xylariaceae Tul. & C. Tul. 1863

ClypeosphaeriaFuckel, Jahrb. Nassauischen Vereins Naturk. 23–24: 117 (1870) [1869–70]

Type species. Clypeosphaeria notarisii Fuckel (a synonym of C. mamillana (Fr.) Lambotte).

Clypeosphaeria is characterised by ascomata with a conspicuous clypeus and brown, oblong, 1-celled ascospores with 3 indistinct pseudosepta and without a germ slit (Munk 1957). Barr (1989) extended the generic concept to include species without pseudosepta, but it is doubtful that the two other species recognised in Clypeosphaeria by Barr (1989), C. perfidiosa and C. americana, are congeneric with C. mamillana, the current name of C. notarisii.

Clypeosphaeria mamillana (Fr.) Lambotte, Fl. Mycol. Belgique (Verviers) 2: 247. 1880 — Fig. 12

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Fig. 12

Clypeosphaeria mamillana. a–g. Ascomata in face view embedded in cleared host tissue, showing the clypeus and ostiole; e, f, k, t, u. ascus apices (e, f, t. in Lugol; k, u. in 3 % KOH + Lugol); g–j, l–q, v–af. ascospores, note the three pseudosepta disappearing in 3 % KOH (ad–af) and the plasma separating from the concave side of dead ascospores (h–j, o, q); r, s. asci with filiform paraphyses (in 3 % KOH) (a, e–j: UPS F-175463 (lectotype of Sphaeria mamillana; images by I. Olariaga); b, c, r–s, ad–af: WU 33598 (epitype of Sphaeria mamillana); d, k–q: Fuckel, Fungi Rhenani 1823 (W 2015-04170, lectotype of Clypeosphaeria notarisii); t–ac: WU 33599). — Scale bars: a, b, d = 0.5 mm; c = 0.2 mm; e–q, t–af = 10 μm; r, s = 20 μm.

Basionym. Sphaeria mamillana Fr., Kongl. Vetensk. Acad. Handl. 39: 103. 1818.

= Clypeosphaeria notarisii Fuckel, Jahrb. Nassauischen Vereins Naturk. 23–24: 117. 1870 [1869–70].

For additional synonyms, see Barr (1989).

Typification. Lectotype of Sphaeria mamillana, here designated: FRANCE, on Cornus sanguinea (C. alba, according to branch colour), Herb. Guepin (UPS F-175463!; MBT202963). Epitype of Sphaeria mamillana, here designated: FRANCE, 21, Dijon, Rue de Malines, on branches of Cornus alba, 28 Aug. 2015, A. Gardiennet AG15060 (WU 33598; MBT202964; ex-epitype culture CLM = CBS 140735). Lectotype of Clypeosphaeria notarisii, here designated: GERMANY, Oestrich, Oestricher Wald, on branches of Rubus fruticosus, without date, L. Fuckel, Fungi Rhen. Exs. 1823 (W 2015-04170!; MBT203104).

Ascomata perithecial, immersed in a distinct clypeate stroma, with a black clypeus 0.5–1.2 mm diam and a central apical papilla, scattered to densely aggregated with confluent clypei, depressed globose to ellipsoid, 400–600 μm diam, circular in transverse section. Apical papilla distinct, black, 50–90 μm diam, commonly surrounded by a lighter coloured zone. Peridium 15–25 μm wide, blackish brown above, lighter brown at the base. Hamathecium of apically free, sparsely branched, 1.3–4 μm wide paraphyses similarly long as the asci. Asci in 3 % KOH (139–)150–174(–181) × (6.5–)7.5–10.5(–12) μm (n = 15), unitunicate, cylindrical, with 8 (partly obliquely) uniseriate, rarely more or less biseriate ascospores, with a short stipe; apex containing a wedge-shaped amyloid apical ring (2.2–)2.5–2.8(–3.0) × (2.3–)2.5–3.2(–3.5) μm (n = 21). Ascospores (13.7–)17.8–21.5(–25.2) × (4.6–)5.3–6.2(–7.2) μm, l/w (2.2–)3.1–3.8(–4.4) (n = 232), ellipsoid, oblong to fusiform, commonly slightly curved, with subacute to rounded ends, light to dark brown, 1-celled and when vital with 3 indistinct pseudosepta and few small guttules, when dead cytoplasma separating from the concave side and forming 4 compartments at the other side, in 3 % KOH pseudosepta becoming faint to invisible. Colonies on MEA and CMD hyaline to pure white, dense, radial, growth radius at 22 °C after 1 mo to c. 50 mm on CMD, to c. 40 mm on MEA. No asexual morph observed.

Habitat — In bark of various shrubs and herbaceous plants; e.g. on Cornus (type host of S. mamillana), especially C. alba including its cultivars; Rubus fruticosus (type host of C. notarisii); Epilobium spp.

Distribution — Europe, probably North America.

Additional material examined. FRANCE, 21, Saint-Apollinaire, D700, L’Arc, on branches of Cornus alba, 28 Aug. 2015, A. Gardiennet AG15061 (WU 33599, culture CLM1); Marcille-sur-Tille, Grand rue, on branches of Cornus alba, 19 Aug. 2015, A. Gardiennet AG15056 (WU 36850, culture CLM2); Marcille-sur-Tille, Rue de Messageries, on branches of Cornus alba, 19 Aug. 2015, A. Gardiennet AG15059 (WU 36851). – GERMANY, Steinberg, on branches of Rubus fruticosus, spring, without date, L. Fuckel, Nassau’s Flora, ex Herb. Barbey-Boissier (W 1904-3852); Oestrich, Oestricher Wald, Mappen, on branches of Rubus fruticosus, winter, without date, L. Fuckel, Herbier Fuckel 1894 ex Herb. Barbey-Boissier (W 1922-11835).

Notes — We treat Clypeosphaeria mamillana here as it is the type of Clypeosphaeriaceae, which has not been included in recent molecular phylogenies. The generic type, Clypeosphaeria notarisii, is generally accepted to be synonymous with C. mamillana, which has priority (for details see Barr 1989). A Guepin specimen of Sphaeria mamillana ex Herb. Fries, which is here designated as lectotype, was examined and documented; its ascospore size (20.2–24.8 × 5–6.2 μm) and septation perfectly fit recent French collections of the species. To stabilise the nomenclature, the recent collection WU 33598, which also originates from France and for which a culture and sequences are available, is here designated as epitype of Sphaeria mamillana. Our investigations of material of C. notarisii collected by Fuckel revealed identical ascospore size and septation, confirming synonymy of Clypeosphaeria notarisii with C. mamillana. The specimen of Fuckel’s Fungi Rhenani 1823 (mentioned in the protologue) preserved in W is here designated as lectotype of C. notarisii. Upon colonisation by Clypeosphaeria, the small branches of Cornus alba and Rubus fruticosus are usually cleared, becoming light brown to whitish, and ascomata are then easy to spot. Barr (1989) reported occasional occurrence of a basal euseptum and small germ pores or slits at the ends of the ascospores, which could not be observed in our investigations. Our phylogenetic analyses places C. mamillana in Xylariaceae (Fig. 1), and classification in a separate family Clypeosphaeriaceae is therefore unjustified.

Clypeosphaeria uniseptata, included as representative of Clypeosphaeriaceae by Senanayake et al. (2015), is not congeneric with Clypeosphaeria but belongs to Lepteutypa (see above).

Xylariales Nannf. 1932 inc. sed.

BasiseptosporaJaklitsch & Voglmayr, gen. nov. — MycoBank MB814844

Etymology. Referring to the basal ascospore septum.

Type species. Basiseptospora fallax (Petr.) Jaklitsch & Voglmayr.

Genus of Xylariales. Perithecia small, immersed in host bark below pallid to brownish patches, scattered, depressed globose, hyaline to yellowish, with a (sub-)hyaline pseudoparenchymatous peridium of thin-walled cells, outermost peridial cell layer becoming brownish with age. Hamathecium of apically free, septate paraphyses embedded in gel. Asci fusoid to clavate, octosporous, unstable when fresh, apex containing an amyloid ring. Ascospores distinctly apiosporous with a small lower cell, straight or slightly curved, surrounded by a gelatinous sheath swelling in 3 % KOH. Asexual morph not known.

Notes — Basiseptospora differs from the unrelated genus Pseudomassaria primarily in its pallid peridium. The phylogenetic relationship to Polyancora, a genus with peculiar condial morphology and ecology (Voglmayr & Yule 2006), is highly supported in ML bootstrap analyses, but receives no significant bootstrap support in MP analyses. The familial affiliation of Basiseptospora is currently unresolved, and we refrain from describing a new family due to lack of significant backbone support of the deeper nodes of the phylogenetic trees.

Basiseptospora fallax (Petr.) Jaklitsch & Voglmayr, comb. nov. — MycoBank MB814845

Basionym. Apiosporina fallax Petr., Ann. Mycol. 25, 3/4: 279. 1927.

Pseudapiospora fallax (Petr.) Petr., Hedwigia 68: 233. 1928.

Pseudomassaria fallax (Petr.) Arx, Ber. Schweiz. Bot. Ges. 62: 349. 1952.

Typification. Lectotype of Apiosporina fallax, here designated: CZECH REPUBLIC, Moravia, Hranice (Mährisch-Weißkirchen), shrubs at the margin of the common pasture of Velká (Welka), on dead twigs of Cornus sanguinea, Oct. 1926, F. Petrak (W 1978-0010762; MBT203101); same collection data (W 1992-0005924, isotype). Epitype, here designated: AUSTRIA, Vienna, 3rd district, Botanical Garden of the University, on corticated twigs of Cornus sanguinea, 9 Jan. 2011, H. Voglmayr (WU 31326, ex-epitype culture CBS 129020 = PSC; MBT203102; ITS-LSU sequence JF440983).

Notes — This species has been described and illustrated as Pseudomassaria fallax in detail by Jaklitsch & Voglmayr (2012). Two collections are mentioned in the protologue, and of the first of these, two specimens are extant in W, one of which is here selected as lectotype. To stabilise the nomenclature, the recent collection WU 31326, which was illustrated in Jaklitsch & Voglmayr (2012) and for which a culture and sequences are available, is here designated as epitype.