Emendation of the genus Tricharina (Pezizales) based on phylogenetic, morphological and ecological data Nicolas VAN VOOREN Uwe LINDEMANN Rosanne HEALY Ascomycete.org, 9 (4) : 101-123. Juillet 2017 mise en ligne le 19/07/2017 Abstract: Tricharina is one of the most difficult genera of Pezizales because it is hard to distinguish morphologically among species. To provide a more robust taxonomy, new investigations on the genus were conducted, both morphologically and phylogenetically. This study focuses on the four key species of the genus: T. ascophanoides, T. gilva, T. ochroleuca and T. praecox. The type material of these species was reviewed. in the literature, T. gilva is considered to be close or even identical to T. ochroleuca. The phylogenetic analysis shows that all recent collections identified as T. ochroleuca appear to be T. gilva. furthermore, the morphological study of the type material of T. ochroleuca leads to the conclusion that the name is a nomen dubium. The presence of several endophytes sequences in the Tricharina-core clade suggests that this genus has an endophytic lifecycle. The confusion between T. gilva and T. praecox, both considered as pyrophilous species, is now resolved thanks to phylogenetic analyses and new data based on vital taxonomy. in contrast to T. gilva, which can occasionally grow on burnt places, T. praecox is a strictly pyrophilous taxon and belongs genetically to a different clade than T. gilva. in agreement with art. 59 of icn, the anamorphic genus Ascorhizoctonia is used to accommodate the species belonging to the “T. praecox clade”. T. ascophanoides, an extra-limital species, is excluded from the genus Tricharina and combined in the genus Cupulina based on the phylogenetic results. The genetic exploration of some other tricharinoid species, i.e. Leucoscypha semiimmersa and L. patavina, confirms they do not belong to the genus Leucoscypha in its original sense, and deserve their own genus. The name Sepultariella is validated to accommodate the latter. Keywords: Ascorhizoctonia, Cupulina, Leucoscypha, Pseudaleuria fibrillosa, Pyronemataceae, ribosomal dna, Scutellinia-Trichophaea lineage, Sepultariella, taxonomy. Introduction The genus Tricharina is a member of Pezizales and was proposed as a new name by Eckblad (1968: 60) replacing Tricharia boud., an illegitimate name. The latter was first published by boudiEr (1885) in his “tribu cupulés, groupe lachnés” for species with short, stiff, brown hairs, longer at the margin, with a sharp top, and smooth, eguttulate ascospores (“sans sporidioles”). Peziza gilva boud. and Lachnea fimbriata Quél. were designated to belong to this genus. boudiEr (1907a) partially emended this genus: it contains species with smooth ascospores or rarely verrucose without oil drops or only with some minute granules, sometimes organized in small groups (“quelques petites granulations parfois réunies en groupes”). These small inclusions are named “bipolar spore granules”, bSG, by kušan et al. (2015). Twenty species were listed. The type-species, Peziza gilva, was designated by clEmEnTS & ShEar (1931: 329), followed by Eckblad (1968) and korf (1972). due to the rather similar characters of the Tricharina species, numerous confusions appeared in the literature and misinterpretations were frequent. for example, the concept of Tricharina gilva by brummElEn (1983) was reevaluated as Wilcoxina mikolae (chin S. Yang & h.E.Wilcox) chin S. Yang & korf (see YanG & korf, 1985b). recently lindEmann (2013) tried to clarify some taxonomical aspects and proposed a new key to the 12 accepted taxa. The genus was monographed by YanG & korf (op. cit.) who proposed a more accessible taxonomy, although the species were studied exclusively from herbarium material. in their work, the genus Wilcoxina was erected to segregate Tricharina mikolae chin S. Yang & h.E. Wilcox due to morphological differences and a different type of asexual stage. Since that publication, two species were combined in Tricharina, T. flava (fuckel) J. moravec (moravEc, 1990) and T. herinkii (Svrček) benkert (bEnkErT, 2010), and three additional new species were published: T. tophiseda matočec & i. kušan (kušan et al., 2015), T. glabra u. lindemann & böhning (lindEmann & böhninG, 2016) and T. aethiopica u. lindemann (lindEmann, 2017). one species has been excluded from the genus: Tricharina fibrillosa (currey) chin S. Yang & korf was transferred to the genus Pseudaleuria as P. fibrillosa (currey) J. moravec (moravEc, 2003). in the present work, our phylogenetic analysis of P. fibrillosa confirms the placement in Pseudaleuria (cf. fig. 1 and 2). Phylogenetic studies on the family Pyronemataceae (PErrY et al., 2007; hanSEn et al., 2013) revealed that Tricharina was paraphyletic. molecular analyses conducted in the course of the study of some tricharinoid species (van voorEn et al., 2015a, 2015b) confirmed the paraphyly. The present work is a contribution to serve the reevaluation of the genus Tricharina in the light of new molecular analyses, the review of type material, and new morphological data based on vital taxonomy methods (baral, 1992). Materials and methods Morphology and cytology. — The observations were made both on fresh and dried material. for dried samples, some small pieces of specimen were rehydrated for about twelve hours in tap water or in potassium hydroxide (5% koh). The following mounts were used to observe microscopic characters: tap water, cotton blue (cb) both in lactophenol and lactic acid, aqueous solution of cresyl blue (crb), 5% potassium hydroxide (koh), and lugol’s solution (iki) or melzer (mlZ) to test the amyloid reaction. measurements were made on 20 to 30 ascospores from each collection, mounted in water, under the 100× oil immersion lens of transmission light microscopes, excluding the ornamentation. “X” represents the average value of spore dimensions, and “Q” the ratio between spore length and width. macrophotographs were made in situ using digital cameras, while micrographs were taken using digital cameras mounted directly on microscopes. line drawings were made freehand to scale. abbreviations: † is used to annotate observations from dead rehydrated material. DNA extraction, amplification and sequencing. — dna was extracted using the same method as described in van voorEn et al. (2015a). loci that were amplified included the iTS (iTS1, 5.8S and iTS2 rdna) and lSu (d1, d2 and d3 of the 28S rdna). Sequences obtained during this study were deposited in Genbank under the accession numbers listed in Table 4. Phylogenetic analyses. — blaST® was used to select the most closely related iTS and lSu sequences from the international nucleotide Sequence database collaboration (inSdc) public databases. These were downloaded and aligned with the sequences obtained during this study using maffT v. 7.187 (kaToh & Toh, 2010), followed by manual adjustment of obviously misaligned columns in Seal v. 2.0a11 (rambauT, 2007). The final iTS alignment had 80 sequences and 475 sites. The lSu alignment had 94 sequences and 847 sites. Ascobolus served as the outgroup for the lSu analyses, while the iTS tree was midpoint rooted. both alignments were analyzed with maximum likelihood (ml) using raxml 8.1.24 (STamaTakiS, 101 2014) with 1000 bootstrap iterations, and the mr. bayes (mb) program (huElSEnbEck & ronQuiST, 2001) using 20,000,000 generations sampled at every 1000 generations with the first 25% of samples discarded as burn in. Priors were estimated in jmodelTest 0.1.1 (PoSada, 2008) using the akaike information criterion. Tracer v1.4 (rambauT & drummond, 2007) was used to inspect the completeness of the mixing of chains, whether stationarity was reached, and the adequacy of sampling. The programs maffT, raxml and mb were run on XSEdE on the ciPrES Science Gateway v. 3.3 (millEr et al., 2010). branch supports that are considered meaningful are shown where raxml bootstrap percentages are ≥ 70%, and mb posterior probabilities are ≥ 0.95. Nomenclature. — all the references to articles of icn come from the melbourne code (mcnEill et al., 2012). new nomenclatural determinations were registered in the mycobank database (http://www.mycobank.org). Results The molecular analyses based on the lSu and iTS loci confirmed that the genus Tricharina is paraphyletic, with species grouped in three distinct well-supported clades (fig. 1 and 2), in the ScutelliniaTrichophaea-lineage (hanSEn et al., 2013). The core clade of Tricharina, which includes the type species Tricharina gilva, is well supported in analyses of both iTS and lSu alignments. The core group includes the following species: T. aethiopica, T. gilva, T. glabra, T. groenlandica, T. hiemalis, and T. striispora. The sequences from the neotype of T. ochroleuca matched those from the holotype of T. groenlandica. The Tricharina-core group is sister to the Geopora lineage. The iTS analyses also revealed the existence of a complex of species around T. gilva. This phylogenetic diversity does not appear in the lSu analyses and we did not find morphological differences in the studied collections to clearly delimit species in this complex. Therefore, T. gilva is here described and illustrated based on its morphological concept. We expect that analyses on other loci could help us to clarify the taxonomic delimitations of these collections. a second clade comprises the Tricharina species around T. praecox and is distinctly outside the Tricharina-core group. The sequenced collections we used — either from Genbank or obtained during this study — suggest that all species belonging to the clade with T. praecox share the same ecology: a habitat on burnt ground. based on these results and supported by morphological characters, we accept the emendation of maTočEc & kušan, in kušan (2015: 55) of the genus Ascorhizoctonia — previously known as an anamorphic genus — for taxa in the T. praecox clade (see discussion in chapter “The Ascorhizoctonia/Tricharina-praecox clade”). a third distinct clade comprised T. ascophanoides and Cupulina montana. We here transfer T. ascophanoides to the genus Cupulina (see discussion in the chapter “The Cupulina-Tricharina-ascophanoides clade”). a new species of Pseudotricharina from argentina was included in these analyses, and will be described in a forthcoming paper. The Tricharina-Core group in this chapter, we focus on the clade containing the type species of the genus Tricharina, Peziza gilva, and allied species. comparisons between the type materials and modern collections gave us the opportunity to better circumscribe some of these species. The iTS and lSu sequences extracted from the type material of P. gilva were essential to the circumscription of the genus Tricharina sensu stricto, which is now strongly supported by molecular evidence. We here provide a description based on all studied and sequenced collections that we recognize as the “true” T. gilva. 1 means “Sarcoscypha”, the subgenus where cooke placed this species. 102 Ascomycete.org Re-examination of type-material of Peziza gilva Peziza gilva boud., in cookE (1879), is typified by the collection Pc 0167813 which contains a single apothecium, in poor condition. The boudier’s label indicates “Lachnea = Tricharia gilva boud. montmorency, terra culta.” contrary to YanG & korf (1985b: 492) who designated this collection as the holotype, there is no evidence in the protologue (cookE, 1879) or in the description provided by boudier in his Icones (boudiEr, 1907b) that can justify the designation as holotype. in our opinion, it seems highly improbable that this taxon was published on the basis of only one element (icn art. 9.1). in the text accompanying plate 347 of Icones mycologicæ, boudier writes “cette espèce, qui n’est pas commune, croît surtout au printemps et en automne […]” which means “This species, which is uncommon, grows mostly in spring and autumn […]”. This suggests that he collected it several times. cooke’s drawing was reproduced from the illustration used by boudier for his plate 347, based on boudier’s find from montmorency. Therefore we consider that this sample is only a part of the protologue and can be only designated as a lectotype. Description (†) of microscopic characters: Excipulum composed of two layers: Medullary excipulum of textura intricata, with hyaline to light yellowish hyphae; Ectal excipulum of textura angularis, composed of brown-yellow cells. Marginal hairs superficial, more or less fasciculate, (80) 115–225 × 9–14 µm, pale brownish, straight, thick-walled, apically obtuse or sharp, with a simple, enlarged base, sometimes subbulbous, septate, wall refractive, 1.2–2 µm thick. Asci cylindrical, about 180–200 µm long, 8spored, base not seen distinctly. Paraphyses not seen. Ascospores uniseriate, ellipsoid, (14.5) 15–17 × 9–10.5 µm [X = 16 × 9.8 µm, n = 40], Q = 1.5–1.8 [Qm = 1.6], sometimes slightly truncate at the poles, smooth, wall up to 0.5 µm thick, without oil drops but containing sometimes bipolar spore granules which merge in koh forming small drops; the wall appears refractive in cb. Comments: boudiEr (1907b) gave a spore size of 13–16 × 9–11 µm which is more in conformity with our re-examination than the dimensions given by cookE (1879), i.e. 14–16 × 7.5 µm, in the protologue. The spores shown on fig. g, plate 347 (boudiEr, op. cit.) are well represented (see fig. 3 and Pl. 3, fig. a-d for comparison), as well as the marginal hairs. indeed, the hair size given by boudier is, after the scale, between 178 and 244 µm in length versus 80–225 µm after our own measurements. Re-examination of type-material of Peziza fimbriata Peziza fimbriata Quél., in cookE (1879), is typified by the collection k(m) 101314 containing two apothecia, in poor condition. The cooke’s label indicates “Peziza (Sarc 1) fimbriata c&Q. on the ground, vosges. france. Quelet”. This collection is undoubtedly the holotype of Peziza fimbriata. Description (†) of microscopic characters: The structure of the excipulum is not clearly discernable, but the ectal excipulum consists of a textura globulosa, with cells 10–32 µm in diam., more elongated in the marginal area, becoming a textura subprismatica. Marginal hairs superficial, fasciculate, 40–137 × 8– 14 µm, brownish, straight, apically obtuse or almost sharp, with a simple, slightly enlarged base, septate, wall not refractive, 1–1.5 µm thick. Excipular hairs superficial, more or less flexuous, 102–355 × 5–6 µm, subhyaline to brownish, with a simple, bulbous base, up to 22 µm wide, 1.5–2 µm thick-walled. Asci 8-spored. Paraphyses not seen. Ascospores uniseriate, ellipsoid, 15–17 × 9–10 (10.2) µm [X = 15.7 × 9.6 µm, n = 22], Q = 1.6–1.8 [Qm = 1.6], sometimes slightly truncate at the poles, hyaline to pale yellowish, smooth, without oil drops, nor bSG, thin-walled, wall refractive in cb. Fig. 1 – raxml consensus phylogram from analysis of lSu alignment of Tricharina and related species, rooted with Ascobolus. meaningful supports shown at branch nodes, with ml bootstrap support on the left and mb posterior probability on the right. branches are thickened where both supports are meaninful: raxml bootstrap support is ≥ 70% and mb posterior probability is ≥ 0.95. Endophyte sequences are green, fruitbody sequences are black. bolded terminals are newly generated sequences. red boxes outlined in yellow indicate pyrophilous fruitbodies. Ascomycete.org 103 Fig. 2 – raxml consensus phylogram of analysis of iTS alignment of Tricharina and related species, midpoint rooted. meaningful supports shown at branch nodes, with ml bootstrap support on the left and mb posterior probability on the right. branches are thickened where both supports are meaninful: raxml bootstrap support is ≥ 70% and mb posterior probability is ≥ 0.95. Endophyte sequences are green, ectomycorrhizal sequences are brown, orchid mycorrhiza sequences are purple, fruitbody sequences are black. bolded terminals are newly generated sequences. red boxes outlined in yellow indicate pyrophilous fruitbodies. 104 Ascomycete.org based on our own experience, marginal hairs of T. gilva can reach exceptionally 300 µm in length, so the boudier’s collection of T. fimbriata could represent T. gilva. on the other hand, we studied a collection from the french Jura (coll. n.v. 2011.08.04) resembling T. gilva but with hairs reaching 420 µm in length, with ascospores 15–16 (17) × 9.8–10.5 µm. This collection appears phylogenetically different from the T. gilva clade and very close or identical to T. hiemalis based on iTS sequences. in this context, the question remains open to know if Peziza fimbriata in its original sense is distinct or not from T. gilva. another interpretation of Peziza fimbriata comes from dEnniS (1978) who suggested this species could be a synonym of Tricharina cretea (cooke) J. moravec. This opinion is shared by moravEc (1974). We don’t think that this hypothesis is correct because of the shape of ascospores drawn by cookE (1878, fig. 362, as Peziza cretea) which is clearly fusoid contrary to the shape of P. fimbriata which is ellipsoid. later moravEc (pers. comm.) examined the type collection of this fungus and revised his position, considering that it is identical with Tricharina gilva. Re-examination of type-material of Peziza ochroleuca Fig. 3 – Peziza gilva. lectotype Pc 0167813. micro-characters. a. ascospores, in water. b. marginal hairs. Scale bars = 10 µm. Comments: Peziza fimbriata Quél., nom. illeg. 2, is the second species cited by boudiEr (1885, under the name Lachnea fimbriata) belonging to the original genus Tricharia. YanG & korf (1985b) synonymized it to Tricharina gilva despite some microscopic differences, especially the length of marginal hairs. The species was described as having marginal hairs reaching about 500 µm in length (cookE, 1879), and ascospores size of 17–18 × 8 µm, although QuélET (1879) gave only 15 µm. after our re-examination, the spore size does not reach the dimensions given by cooke and is closer to the “length” given by Quélet. as noted by YanG & korf (1985b), cooke probably measured overmature ascospores. We did not find any long hair on the sample but it is in poor condition. The hairs observed were short and not different from those seen on the type specimen of T. gilva. boudiEr (1907b) also illustrated this species under the name Tricharia fimbriata with another collection from eastern france with hairs reaching 290 µm in length and spore size 16–17 × 10 µm. Peziza ochroleuca bres. is lectotypified by the collection S-f190915 which is considered as part of the type material but contains very little material (mostly substrate), in poor condition, and we were unable to locate any portion of an apothecium. The bresadola’s label indicates “Lachnea ochroleuca bres. 3 in silva conifera Tertiolasii. Sp. 15-18 = 10-12. G. bresadola”. This mention agrees with the protologue in brESadola (1887). YanG & korf (1985b: 500) examined this collection (“we use the characters associated with that apothecium as a basis for a concept for bresadola’s name”), but surprisingly decided to designate a neotype from a more abundant norwegian collection. This is not in accordance with the current nomenclatural rules concerning the typification of names (icn art. 9). after examining the collection Sf190915 we consider this lectotype as totally “unusable”. in this context, the plate civ, fig. 2 in bresadola’s Fungi tridentini novi, cited in the protologue (brESadola, 1887) should serve as a lectotype. another little packet is present in the envelope and contains a single apothecium glued to a slip of paper. it was separated by YanG & korf (1985b) and considered to be a Cheilymenia species. The examination of this sample reveals ellipsoid ascospores, with tapered ends, eguttulate, measuring 15–18 × 10–11 µm. in cb, spores show a yellow refractive content — which is typical of the genus Cheilymenia — and are finely ornamented with low warts, partially elongated and connected, sometimes mixed with pustulate warts, as some species of the section Pseudoscutelliniae (moravEc, 2005: 171). unfortunately we did not see any hair. Strangely the spores drawn by brESadola (1887, Pl. civ, fig. 2) look like those we observed in this Cheilymenia sample 4 and the hairs also resemble to hairs of this genus. YanG & korf (1985b) indicated about this drawing: “his hairs are clustered but too sharp for Tricharina.” So it seems that bresadola mixed up two species. at the same time, our phylogenetic analyses show that iTS sequence JQ824126 from a culture based on this neotype of Tricharina ochroleuca (YanG & korf, 1985b) is identical to the iTS sequence JQ824125 from a culture based on the holotype of T. groenlandica. The lSu sequences (JQ836561 and u38576) are also identical. These results suggest that the neotype designated by YanG & korf (op. cit.) for T. ochroleuca must be rejected. for all these reasons, we propose to consider Peziza ochroleuca bres. as a nomen dubium and to abandon the name Tricharina 2 not Peziza fimbriata chaillet ex fr., Syst. mycol., 2 (2): 511 (1823). The name Lachnea fimbriata Quél. (1879) is considered as a nomen novum. 3 This name was not retained in brESadola’s (1887) original description. The name P. ochroleuca choosen by bresadola is illegitimate due to Peziza ochroleuca bolton (1789), an older valid name. The name Lachnea ochroleuca proposed by Saccardo (1889) has to be considered as a nomen novum (icn art. 58.1). 4 in his diagnosis, bresadola qualifies the spores as “subhyalinae”, maybe due to an effect of refractiveness. Ascomycete.org 105 A B C Plate 1 – Peziza fimbriata. holotype k(m) 101314. micro-characters. a. ascospores in 5% koh. b. marginal hairs, in water. c. bulbous hairs from the excipular surface, in water. Scale bars = 10 µm. ochroleuca (bres. ex Sacc.) Eckblad because the concept introduced by YanG & korf (op. cit.) is doubtful. Other species in the Tricharina-Core group other species that share the well-supported clade with Tricharina gilva are: • Tricharina aethiopica u. lindemann, a well-defined species collected in Ethiopia which has many similarities with T. gilva but differs in very long hairs and the bright yellowish-brown color of the hymenium (lindEmann, 2017). for the time being it is only known from the type locality. • T. glabra u. lindemann & böhning, a well-defined species, characterized by very short hairs, broad ellipsoid spores with many fine 106 Ascomycete.org oil droplets inside and a positive iodine reaction (greenish) of the orange-brown content of the paraphyses. in regard to the morphology, this species differs the most from the generic concept of Tricharina as outlined by YanG & korf (1985b). until now, this species was known only from the type locality on the island fehmarn in the baltic Sea. • T. groenlandica dissing, chin S. Yang & korf, characterized by excipular and marginal short hairs, intermixed with undifferentiated brown cells fide YanG & korf (1985b), distributed in northern Europe and northern america; until now there is no collection of these species known which was described using methods of vital taxonomy. Therefore morphological studies of fresh material are required to have a more complete image of this taxon. • T. hiemalis chin S. Yang & korf is a widespread but rare species, known from north america (WhiTnEY & ParmETEr, 1964) and Europe (YanG & korf, 1985b). it is very close to T. gilva but the spore shape is slightly different, the marginal hairs are distinctly longer than those of T. gilva (reaching up to 460 µm), and the apothecia of T. hiemalis are generally more hairy than those of T. gilva. because of the slight differences with T. gilva (as far as we know them), further morphological studies of fresh material are required. • T. striispora rifai, chin S. Yang & korf, a well-defined species, characterized by striate ascospores (YanG & korf, 1985b; arGaud, 2008; Galán et al., 2010; ivaldi & fouchiEr, 2014; SammuT, 2016). This is a widespread species known from australia (type locality), Southern Europe (Portugal, Spain, france, Greece, malta) and South america; it is often found in or near to plantations of Olea europaea. The phylogenetic positions of T. japonica chin S. Yang & korf and T. tophiseda matočec & i. kušan are unknown. based on the morphology and ecology of these species we predict that they are most closely affiliated with species in the Tricharina-core group. The same could be stated for T. pallidisetosa (E.k. cash) k.S. Thind & S.c. kaushal and T. flava (fuckel) J. moravec which are known only from their type localities. descriptions of T. herinkii (Svrček) benkert in SvrčEk (1949) and bEnkErT (2010) morphologically situate this species between Tricharina s. str. and Geopora harkn. s. l. our review of Senn-irlet’s collection of Geopora herinkii (SEnn-irlET, 1989) proved to be T. gilva, both morphologically and phylogenetically. further studies are required, especially in regard to the phylogenetic positions of these three taxa. Taxonomy Tricharina gilva (boud.) Eckblad, Nytt Mag. Bot., 15 (1-2): 60 (1968). basionym: Peziza gilva boud., in cooke, Mycographia, 6: 240 (1879). lectotype here designated: Pc 0167813; mbT 204678; Genbank kY364019 (iTS) and kY364057 (lSu). homotypic synonyms: Humaria gilva (boud.) Sydow, Mycoth. march. no. 775 (1884); Lachnea gilva (boud.) Sacc., Syll. fung., 8: 184 (1889); Scutellinia gilva (boud.) kuntze, Rev. gen. pl., 2: 869 (1891); Tricharia gilva (boud.) boud., Icon. Mycol., liste prelim.: 3 (1904); Patella gilva (boud.) Seaver, N. Amer. Cup-Fungi (Operculates): 166 (1928); Trichophaea gilva (boud.) Gamundí, Rev. Mus. La Plata, 10: 60 (1967). Taxonomic synonyms: Ascorhizoctonia gilva chin S. Yang & korf, Mycotaxon, 23: 472 (1985). for other putative synonyms, see YanG & korf (1985b). Apothecia cupulate, 5–6.5 mm in diam., partially buried in the substrate, sessile, hymenium pale greyish to ochraceous grey, dull orange; external surface concolorous or darker, finely pustulate, co- vered by scattered, hyaline hairs. Margin eroded, covered by small brown hairs, often fasciculate. Excipulum composed of two layers, a thin medullary excipulum, of textura intricata, composed of hyaline hyphae, and an ectal excipulum of textura globulosa to subangularis, with cells 10–40 µm wide, hyaline to yellowish brown in the outer part. Excipular hairs superficial, scattered, more or less flexuous, 150–370 (550) × 5– 6.5 µm, brownish yellow at the base, hyaline in the upper part, with a simple base, septate, wall up to 0.5 µm thick. Marginal hairs superficial, straight, 60–260 (300) × 9–10 µm, pale brownish, often subhyaline at the top, apically obtuse for the smallest ones, more or less sharp for the other, with a simple base, septate, wall up to 1 µm thick. Asci cylindrical, 160–190 × 12–16 µm, with crozier, 8-spored, inamyloid. Paraphyses not enlarged at the top, 3–5 µm in diam., hyaline, septate, without vacuoles. Ascospores 5 ellipsoid with rounded ends, 14–17.5 (18.1) × (8) 8.5–11 (11.5) µm [X = 16 × 9.9 µm], Q = 1.4–1.8 [Qm = 1.6], † (14) 14.5–17 × (8.5) 9–10.5 µm [X = 15.8 × 9.7 µm], † Q = 1.5–1.8 [Qm = 1.6], sometimes very slightly truncate (mostly in rehydrated exsiccates, when stained in cotton blue with lactic acid [see Plate 3, a2] or when fresh but a bit overmature [see Plate 3, d1], hyaline, smooth, thin-walled when mature, slightly thick-walled when unripe or overmature, mature without oil drops but when overmature with many very minute droplets due to the process of germination (lindEmann, 2013, fig. 5), rarely with bipolar spore granules. Studied collections: cZEch rEPublic – central bohemia, Prague, 50.084444° n 14.423888° E, alt. 197 m, in a huge flowerpot among Tagetes, leg. Z. Egertova, 17 Jul. 2014, pers. herb. u.l. 241. francE – doubs, levier, route forestière de ravonnet, 46.929224° n 6.080289° E, alt. 750 m, on soil, in a rut, leg. n. van vooren, 17 aug. 2011, pers. herb. n.v. 2011.08.05. bouches-du-rhône, marseille, parc longchamp, 43.30575° n 5.397855° E, alt. 63 m, on soil, leg. m. vega, 20 mar. 2014, pers. herb. n.v. 2014.03.11. GErmanY – Erbach/donau, 48.326091° n 9.898097° E, alt. 480 m, on sandy, clay-like soil within bark mulch at a parking place of a supermarket, leg. r. Seibert, 26 apr. 2014, pers. herb. u.l. 191. SPain – la rioja, Sojuela, 42.356389° n -2.55° E, alt. 805 m, on sandy and moist soil on the bank of a river, between Corylus avellana, Castanea sativa, Populus sp. and Quercus sp., with Verpa conica, Peziza depressa and Pachyella violaceonigra, leg. rubén martinéz Gil, 23 apr. 2016, pers. herb. u.l. 243. asturias, Pola de Somiedo, near the school, 43.090328° n -6.257085° E, alt. 718 m, on soil, leg. G. moyne, 2 Jun. 2013, pers. herb. n.v. 2013.06.00. la rioja, San millán, río cárdenas, 42.275555° n -2.921944° E, alt. 940 m, on an old burnt site, leg. rubén martinéz Gil, 6 dec. 2012, pers. herb. r.m.-1254. SWiTZErland – valais, massif of Great St. bernard, bourg-Saint-Pierre, valsorey, Sander des Glaciers de valsorey, 45.917722° n 7.259806° E, ca. alt. 2390 m., leg. and det. b. Senn-irlet, 27 aug. 1983, herbarium helveticum b. irlet no. 83/168, under the name Geopora herinkii. Table 1 – range of spore size and marginal hair size of the collections of Tricharina gilva sequenced during this study Spore size (μm) Mean spore size (μm) Q value Mean Q value Marginal hair size (μm) Pc 0167813 (lectotype) † (14.5) 15–17 × 9–10.5 (n = 40) 16 × 9.8 1.5–1.8 1.6 up to 225 n.v. 2011.08.05 14–16.5 × (8.5) 9–10.5 (n = 20) 15.2 × 9.6 1.5–1.8 1.6 up to 155 Coll. Number n.v. 2013.06.00 15–17 × 9–10 (n = 20) 15.9 × 9.5 1.5–1.8 1.7 up to 170 14.5–18 × 9.3–11.9 (n = 36) 16.3 × 10.6 1.4–1.7 1.6 up to 300 15.5–17.3 (18.1) × 9.1–10.5 (n = 24) 16.7 × 9.9 1.6–1.8 1.7 up to 260 u.l. 241 15.8–17.8 × 10.1–11 (11.5) (n = 15) 16.6 × 10.9 1.4–1.6 1.5 up to 280 u.l. 243 15–17.5 (18.4) × (8.6) 9.3–11 (n = 49) 16.3 × 10.2 1.5–1.7 1.6 up to 200 herb. helveticum no. 83/168 † (14) 15–16.3 × 8.5–9.6 (9.9) (n = 14) 15.4 × 9.2 1.6–1.7 (1.8) 1.7 up to 280 r.m.-1254 u.l. 191 5 for the data of the all sequenced collections see Table 1. Ascomycete.org 107 Plate 2 – Tricharina gilva. macroscopic aspects. a. coll. u.l. 191. b. coll. n.v. 2013.06.00. c. coll. n.v. 2011.08.05. d. coll. u.l. 241. E. u.l. 243. f. boudier’s drawing of Tricharia gilva adapted from his Icones pl. 347. G. coll. r.m.-1254. 108 Ascomycete.org Discussion in the light of our results, Tricharina gilva must be regarded as mostly non-pyrophilous contrary to the opinion of several authors (Eckblad, 1968; maaS GEESTEranuS, 1969; dEnniS, 1978; diSSinG et al., 2000; mEdardi, 2006; bEuG et al., 2014; van voorEn, 2014). although T. gilva may exceptionally occur on burnt ground (see collection r.m.-1254), all other studied collections that were collected from a fire place and initially identified as T. gilva were phylogenetically inferred in the T. praecox clade. The misinterpretation of T. gilva as a pyrophilous taxon (strictly or occasionally) can be explained by the confusion over species and generic delimitation, as apparent in the “Ascorhizoctonia/Tricharina-praecox” clade of the phylogenetic trees (fig. 1 and 2) where extra-generic species have been determined under this name. The ecology of the species in the Tricharina clade appears to include an endophytic habit. There is also evidence that some species play a role in orchid mycorrhizae in both hemispheres. The subtle morphological differences between T. gilva and T. praecox has led to many misidentifications. The color of living apothecia can possibly be used to differentiate the species. it seems that apothecia of T. praecox are more “yellowish” to “orange” in the fresh state (see Plate 5), probably due to a yellowish pigment present in the paraphyses, which is more or less dense and visible in the living state; this pigment is always lacking in T. gilva. microscopically, the main difference comes from the spore shape: in T. gilva, the ascospores are mainly ellipsoid with rounded ends; in T. praecox, they are mainly ellipsoid with tapered ends (looking more fusoid, see Plate 3, E-i). Tricharina ochroleuca (bres. ex Sacc.) Eckblad is another name often used for collections corresponding in fact to T. gilva. as T. gilva was generally accepted as a pyrophilous species in modern literature, we think that many mycologists named their collections of brownish or ochraceous Tricharina growing on soil under the epithet ochroleuca. although boudiEr (1907b), Eckblad (1968) and YanG & korf (1985b) considered this taxon as a good species, their interpretations seem different. as shown in our phylogenetic analyses, the neotype of T. ochroleuca, chosen by the latter authors, is in reality conspecific with T. groenlandica. as previously said, we consider Peziza ochroleuca bres. (and the subsequent homotypic synonyms) as a nomen dubium. in the literature, the range of spore size of T. gilva is heterogeneous (see Table 2). Surprisingly rEhm (1895, as Lachnea gilva) gives 10–12 × 7–8 µm, a range value too small to fit T. gilva. The revision of this collection by YanG & korf (1985b) demonstrated this was a new species, published by them as Wilcoxina rehmii. SEavEr (1928, as Patella gilva) was the first author who described a collection on an old burnt site determined as T. gilva. GrElET (1939) gives a description in accordance with the boudier’s description, except for the larger marginal hairs that reach 400 µm in length. his collection may represent “Tricharia” fimbriata (see discussion below). The cited Grelet’s collection from nov. 1924 was also revised by YanG & korf (1985b: 527) and identified as Trichophaea gregaria (rehm) boud. SvrčEk (1948) described two collections on burnt places, and the description and the spore drawing (opt. cit., Pl. i, fig. 10) fit for T. gilva, but we cannot exclude the possibility that this represents T. praecox. note that the max length value of 19.5 µm was probably measured on ascospores which are ready to germinate and are starting to swell (cf. lindEmann, 2013: 33, fig. 5). as a general matter, we think that a spore size over 18 µm indicates probably these swollen ascospores. This could be also the case for the collection(s) studied and illustrated by maaS GEESTEranuS (1969). dEnniS (1977) described T. gilva after a collection on a burnt site, but there are few differences with his description of T. praecox, except larger spores for T. gilva. The collection illustrated by brEiTEnbach & kränZlin (1984) represents undoubtedly T. praecox due to the substrate and spore shape; YanG & korf (1985b) who revised the material depicted in breitenbach & kränzlin considered it as T. praecox var. cretea. cETTo (1987) illustrates also a collection on a fire place which could be T. praecox (general aspect and narrow spores). in Nordic Macromycetes (diSSinG et al., 2000), T. gilva is noted as a pyrophilous species with hair length reaching 600 µm! it seems that T. gilva is given here in a very broad sense. a collection from argentina on a burnt site, provided by Gamundí & lorEnZo (2001), is not illustrated but the authors refer to brummElEn (1983) to compare with their fungus. it should be noted, however, that brummelen’s interpretation of T. gilva was reevaluated as a Wilcoxina species (YanG & korf, 1985b). We suppose that the collections illustrated by mEdardi (2006) and bEuG et al. (2014) on pure charcoal, with narrow ascospores might also represent T. praecox instead of T. gilva. after revision, the collection identified by van voorEn (2014) as T. gilva corresponds also to T. praecox. The Ascorhizoctonia/Tricharina-praecox clade as shown in the phylogenetic analyses, Tricharina praecox s. lato belongs to a distinct clade outside the Tricharina-Geopora-lineage. Therefore, a new genus name is required for the species in this clade. although no sequence of the type material of Peziza praecox P. karst. is available, the asexual stage of Tricharina praecox var. praecox has been identified as Ascorhizoctonia praecox chin S. Yang & korf (YanG & korf, 1985a) and chosen as the type-species of the anamorphic Table 2 – range of spore size and marginal hair size found in the literature for collections named Tricharina gilva Reference Correct name rEhm (1895) Wilcoxina rehmii SEavEr (1928) Ascospores size (μm) Hairs (μm) 10–12 × 7–8 150–200 × 6–10 14–18 × 8–10 up to 200 × 8 GrElET (1939) Trichophaea gregaria 15–17 × 10–11 100–400 × 8–12 SvrčEk (1948) Tricharina gilva ? 15–19.5 × 9–11 80–200 × 5.5–8.5 13–16 × 9–11 ? maaS GEESTEranuS (1969) Tricharina gilva ? 17.7–18.5 × 9.5–11 up to 265 × 4–10 dEnniS (1978) Tricharina gilva ? 14–19 × 9–10 up to 200 × 10 brEiTEnbach & kränZlin (1984) Tricharina praecox 14–16 × 8–10 150–190 × 3–5 (10) cETTo (1987) Tricharina praecox ? 14–16 × 8–10 ? diSSinG et al. (2000) Wide concept of T. gilva 16–17 × 8.5–10 200–600 Gamundí & lorEnZo (2001) Tricharina sp. 14–15.5 × 9.5–10 110–230 × 5–7.5 mEdardi (2006) Tricharina praecox ? 14–14.5 × 8.5–10 ~ 200 bEuG et al. (2014) Tricharina praecox ? 14–19 × 8–10 150–200 × 3–5 van voorEn (2014) Tricharina praecox 14–16.5 (17) × (8) 9–10.2 130–280 × 5–10 moSEr (1963) Ascomycete.org 109 Plate 3 – ascospores of Tricharina gilva (left column: a-d) versus T. praecox (right column: E-i) a: T. gilva: coll. u.l. 191; a1 + a3: ascospores in h2o (in the living state); a2: ascospore (with slightly truncate poles), stained in cb in lactic acid. b: T. gilva: coll. u.l. 243; b1 + b3: ascospores in h2o (in the living state); b2: ascospore, stained in cb in lactic acid. c: T. gilva: coll. u.l. 241; c1 + c2: ascospores in h2o, slightly thick-walled (in the living state); d: T. gilva: coll. r.m.-1254; d1 + d2: ascospores in h2o (in the living state), sligthly overmature. E: T. praecox: coll. u.l. 100; E1 + E2: ascospores in h2o (in the living state) E3: ornamentation of the ascospores; f: T. praecox: coll. n.v. 2016.03.01: f1: ascospores in h2o (in the living state), f2: idem, sligthly overmature. G: T. praecox: coll. n.v. 2010.05.32; G1 + G2: (rehydrated) ascospores in h2o (in the dead state). h: T. praecox: coll. dEd 7356 (SfSu); h: (rehydrated) ascospores in h2o (in the dead state). i: T. praecox: lectotype, collection P.a. karsten no. 1176 (h); Scale = 10 µm (except for a3, b3, c2, d2, E2, E3 + f1: scale = 5 µm). 110 Ascomycete.org genus Ascorhizoctonia chin S. Yang & korf. in this context, the icn art. 59 leads the choice for a new name of the genus: Ascorhizoctonia must be used following the principle of nomenclatural priority after the recommendations made by roSSman (2014). The same conclusion was retained by maTočEc & kušan (in kušan, 2015: 55). consequently, all other taxa with similar morphological characters belonging to this clade must be combined in this genus. Re-examination of type-material of Peziza praecox Peziza praecox P. karst. was lectotypified by YanG & korf (1985b) with the collection P.a. karsten no. 1176 housed in the university of helsinki botanical herbarium. it contains some apothecia embedded in black ashes, in rather good condition. The label indicates “Peziza praecox karst. fennia. Tavastia australis. Tammela, mustalia. in carbonario versus haarankorpi.” This agrees with one of the localities cited in the protologue (karSTEn, 1869). Description (†) of microscopic characters: Excipulum rather thin, about 150 µm thick, bilayered, with an ectal excipulum of textura angularis/subglobulosa, with yellowish cells 10–22 µm wide, and a medullary excipulum of textura intricata, with hyaline hyphae. Anchor hyphae present, 4–6 µm wide, hyaline, septate, with a refractive wall, 0.5–1 µm thick, enlarged at the base or subbulbous. Marginal hairs superficial, dense, 112–340 × 9–13 µm, yellow-brown to subhyaline, straight, apically sharp, more rarely obtuse, with a simple, slightly enlarged base, septate, wall not refractive, 1–1.2 µm thick. Asci 8-spored, inamyloid. Paraphyses cylindrical, 2.5–3.5 µm in diam., hyaline. Ascospores uniseriate, ellipsoid, with somewhat tapered poles, (14) 14.5–16.5 (17) × (7.5) 8.7–10 µm [X = 15.7 × 9.4 µm, n = 40], Q = 1.5–1.9 [Qm = 1.7], sometimes slightly truncate at the poles, hyaline, smooth, without oil drops, nor bSG, but with a spumous content, wall thin, up to 0.5 µm, appearing refractive in cb. Comments: The re-examination of this type-collection confirms the main micro-characters we observed on fresh collections, especially the spore shape, the most important element to distinguish Ascorhizoctonia praecox from Tricharina gilva. The spore length is a bit longer than reported by YanG & korf (1985b: 504). The hairs can reach 340 µm in length. it is also important to note that the ascospores are totally smooth. Taxonomy Ascorhizoctonia praecox chin S. Yang & korf, Mycotaxon, 23: 475 (1985). holotype: cuP 61609, ex type culture cbS 240.85; Genbank JQ836562 (28S) and JQ836559 (iTS). Taxonomic synonyms: Peziza praecox P. karst., Not. Sällsk. Fauna Fl. Fenn. Förhandl., 10: 124 (1869); Lachnea praecox (P. karst.) Sacc., Syll. fung., 8: 183 (1889); Scutellinia praecox (P. karst.) kuntze, Rev. gen. pl., 2: 869 (1891); Tricharia praecox (P. karst.) boud., Icon. Mycol., liste prélim.: 3 (1904); Tricharina praecox (P. karst.) dennis, Kew Bull., 25 (2): 338 (1971). Apothecia cupulate, 2–5 mm in diam., sessile, spread at the end, hymenium first greyish on very young specimens, then ochraceous yellow, sometimes with dull orange tinges, sometimes bright yellow, often with some olivaceous tinges; external surface concolorous, covered by small brown hairs, scattered. Margin darker by the presence of small hairs. Excipulum thin, 80–200 µm wide, bilayered, with a medullary excipulum of textura ± intricata to subprismatica, composed of pale orange hyphae, becoming a textura prismatica in the marginal area, and an ectal excipulum of textura subglobulosa, with roundish or widely clavate cells, 15–43 × 15–26 µm, hyaline. Anchor hyphae present, more or less flexuous, 5–8 µm wide, hyaline, smooth, septate, ~1 µm thick-walled, with a simple, bulbous base. Marginal hairs superficial, 150–270 (340) × 7–10 (12) µm, often fasciculate, straight, apically obtuse, with a simple subbulbous base, subhyaline to pale brown, septate, wall refractive, up to 1 (2) µm thick. Asci cylindrical, 200–220 × 11–14 µm, slightly narrowed at the base, with crozier, 8-spored, inamyloid. Paraphyses hyaline, septate, more or less constricted at the septa, not or slightly enlarged at the top or sometimes with a sort of lateral protuberance, 4–5 µm in diam., frequently forked at the apex, containing non-refractive vacuoles, sometimes also a slightly yellowish pigment, partially staining in crb, without reaction in lugol. Ascospores uniseriate, ellipsoid with somewhat tapered poles, 14–18 (19) × (8) 9–11.5 (12.5) µm [X = 16.0 × 10.1 µm], Q = 1.5–1.8 [Qm = 1.6], † (14.5) 15–17 (18) × (8.7) 9–10.5 (11) µm [X = 15.9 × 9.7 µm], † Q = 1.5–1.9 [Qm = 1.7], rarely truncate at the poles, hyaline, without oil drops, but with bipolar spore granules, one or two bigger than the others, not staining in crb, disappearing in koh, smooth or very finely warted (coll. u.l.100) or sometimes distinctly verrucose (coll. r.d. 15.06.245.91), wall up to 0.5 µm thick, appearing refractive in cb. Studied collections: francE – ardennes, Givet, fort condé, 50.13772° n 4.80067° E, alt. 170 m, on a burnt place, under Pinus nigra, leg. b. clesse, 9 mar. 2016, pers. herb. n.v. 2016.03.01. Savoie, méribel-les-allues, near the “altiport”, 45.4104851° n 6.5788434° E, alt. 1760 m, on a recent burnt place, with Peziza lobulata, under Picea abies, leg. n. van vooren, 23 may 2010, pers. herb. n.v. 2010.05.32. doubs, Gevresin, carrefour du roi de rome, 46.9626° n 6.063534° E, alt. 690 m, on a burnt place, leg. G. moyne & n. van vooren, 30 aug. 2006, pers. herb. n.v. 2006.08.27. haute-Savoie, Saint-Jean-de-Sixt, forêt des Traversiers, 45.9163471° n 6.4045474° E, alt. 1010 m, on a burnt place, leg. n. van vooren, 23 Sept. 2006, pers. herb. n.v. 2006.09.21. loire, écoche, bois du châtelet, 46.161544° n 4.3099131° Plate 4 – Peziza praecox h 6055460 lectotype. ascospores. a. ascospores, in water. b. ascospores in cb, showing their refractiveness. Scale bar = 10 μm. Ascomycete.org 111 Plate 5 – Ascorhizoctonia praecox. macroscopic aspects. a. coll. n.v. 2016.03.01. b. coll. r.d. 15.07.245.91. c. coll. fellmann 2007. d. coll. n.v. 2010.05.32. E. u.l.100. f. boudier’s drawing of Tricharia praecox adapted from his Icones pl. 349. 112 Ascomycete.org A B C D Fig. 4 – Ascorhizoctonia praecox. coll. n.v. 2016.03.01. micro-characters. a. ascospores, in water. b. Top of ascus and paraphyses. c. anchor hypha, with bulbous base. d. marginal hairs. Scale bars = 10 µm. Ascomycete.org 113 E, alt. 600 m, on a 1-year old burnt area, with Peziza lobulata and P. echinospora, leg. n. van vooren, 18 oct. 2003, pers. herb. n.v. 2003.10.12. finland – Tavastia australis 6, Tammela, mustalia, near haarankorpi {estimated coordinates: 60.8540451° n 23.7764637° E, alt. 120 m}, in ash on burnt ground, leg. P.a. karten, 1 Jun. 1868, h 6055460, ex herb. P.a. karsten no. 1176, lectotype of Peziza praecox. Tavastia australis, Tammela, letku, on burnt ground, leg. P.a. karten, 22 Jun. 1868, h 6055461, ex herb. P.a. karsten no. 2501, syntype of Peziza praecox. SPain – canary islands, la Gomera, las hayas, valle Gran rey, 28.12720723° n -17.2825993° E, alt. 1100 m, in a very wet place, recently burned site, “fayal-brezal” habitat (Myrico fayae-Ericion arboreae), with Ascobolus carbonarius and Peziza sp., 13. feb. 2013, leg. r. negrín, det. m. ribes, conf. u. lindemann, pers. herb. u.l.100. GErmanY – munich, Waldperlach, 48.091415° n 11.675750° E, alt. 550 m, on an old burnt site, leg. b. fellmann, 04. Jun. 2007 (only macro- and micropictures, no exsiccate). SWiTZErland – fribourg, bulle, cabane d’inson, 46.584153° n 7.0010782° E, alt. 1020 m, on a 2-years old burnt site, leg. n. van vooren, 5 Jun. 2015, pers. herb. n.v. 2015.06.09. fribourg, montagny, forêt de la chanéaz, 46.80083° n 7.004722° E, alt. 590 m, on a burnt place, leg. r. dougoud, 29 apr. 1991, pers. herb. r.d. 15.06.245.91, under the name Tricharina praecox var. cretea. uSa – california, calavares county, near big Trees State Park, 38.277573° n -120.308335° E, alt. 1435 m, on burnt soil under conifers, leg. d.E. desjardin, rev. n. van vooren, 20 apr. 2002, coll. dEd 7356 (SfSu) as Tricharina gilva. Discussion as shown previously, a great confusion exists between Tricharina gilva and Ascorhizoctonia praecox due to their similar morphology and the suggestion by the older literature that both species should grow in the same habitat: burnt site or charcoal. contrary to A. praecox which seems strictly pyrophilous, T. gilva grows only occasionally on burnt sites and we think on old ones. a review of numerous herbarium collections should validate these hypotheses. The distinction between T. gilva and A. praecox is easily observed in their different spore shapes, which offer a quick means of identification (see Plate 3). at the same time, this distinction is generally not correctly indicated or drawn in the literature, e.g. dEnniS (1971), so we encourage the readers to review their own material to check their previous determinations. note that A. praecox is probably one of the most common tricharinoid discomycetes on recent fire places (up to 2 years old). The Ascorhizoctonia clade contains collections with multiple determinations. Two of these collections correspond to Tricharina gilva (coll. hd rana 81.118 from herb. c, on “moist soil”), and T. cretea (coll. rd 15.06.245.91, on burnt place). The latter has been revised and corresponds to the fungus called T. praecox var. cretea by YanG & korf (1985b). This taxon differs mainly from T. praecox var. praecox by its distinctly verrucose fusoid ascospores. but phylogenetically, this collection appears to be the same as the collections of A. praecox sequenced during this study. if the dna data from the type collection of Peziza cretea cooke confirm this result, T. cretea should be considered as a synonym of A. praecox. YanG & korf (1985b) proposed another variety of T. praecox called T. praecox var. intermedia Egger, chin S. Yang & korf, based on some north american collections. This taxon is distinguished by Yang & korf from T. praecox var. praecox by its smooth ascospores, a different asexual state and its geographical distribution. our phylogenetic analysis shows that a collection from california (dEd 7356), initially determined as T. gilva, was in fact genetically identical to A. praecox (see fig. 1). our study of the sample of dEd 7356 confirmed this observation also in regard to the microcharacters of this collection: the ascospores were smooth with tapered poles (see Plate 3, fig. h). contrary to the opinion of YanG & korf (1985b) who suggested that A. praecox var. praecox was not present in north america, we think that the situation is more complex. further investigations using the methods of vital taxonomy and phylogeny with additional genetic loci are required to evaluate whether Ascorhizoctonia intermedia could be distinguished by its morphological characters from A. praecox or not. The Cupulina-Tricharina-ascophanoides clade among the species of the genus Tricharina, T. ascophanoides (boud.) chin S. Yang & korf is quite atypical because of the absence of true marginal hairs. on the other hand, its eguttulate ascospores contain bipolar spore granules, a character that agrees well with the typical spore-morphology of the genus as defined by boudiEr (1885) and others. but despite this, the phylogenetic analyses show definitely that T. ascophanoides does not belong to the Tricharina-core clade. Re-examination of type-material of Tricharia ascophanoides Tricharia ascophanoides boud. is typified by the collection Pc 0167814, considered as the holotype. it contains some apothecia in rather good condition embedded in the substrate. The boudier’s label indicates “Tricharia ascophanoides b. vid. icones n° 586. arbois augusto 1902 ad corium putridum legit dr hetier.” These data agree with the protologue (boudiEr, 1903). Description (†) of microscopic characters: The excipulum is bilayered, with a medullary excipulum of textura intricata, composed of hyaline to light yellowish hyphae, and a thin ectal excipulum of textura angularis. Hairs not seen but anchor hyphae present, hyaline, 6–9 µm wide, more or less obtuse at the top, septate, with refractive wall, 1–1.2 µm thick, arising from a bulbous basal cell, 17–26 µm wide. Asci cylindrical, 8-spored, inamyloid. Paraphyses not seen. Ascospores uniseriate, ellipsoid or ellipsoid with somewhat tapered poles, 17–20 × 10–12 (12.7) µm [X = 18.8 × 11.3 µm, n = 30], Q = 1.5–1.9 [Qm = 1.7], sometimes slightly inequilateral, hyaline, without oil drops but with bipolar spore granules that merge in koh, smooth, thin-walled, wall sometimes refractive and pale yellow in cb. Comments: These characters are in conformity with the description published by boudiEr (1903), especially the spore size which is evidently larger than indicated by YanG & korf (1985b: 486) who gave 14.6–16.1 (16.8) × 9.5–11 µm based on the study of only two collections of T. ascophanoides. Such a difference is difficult to explain but probably caused by the medium used to observe the small number of examined material, including the type-collection. our own collections, studied in living state (as boudier did), show that these dimensions are clearly underestimated. We measured (16) 16.5–22.5 (24) × (9.5) 10–13 (14.5) µm with a mean value of 18.7 µm in length and of 11.3 µm in width. YanG & kriSTianSEn (1989) obtained an anamorph in culture — named Ascorhizoctonia ascophanoides by the authors — based on kristiansen’s collection (rk 82.146), with spores measured to 14–16 × 9–11 μm after kriSTianSEn (2014). molecular analyses based on iTS and lSu sequences of the type-material of A. ascophanoides show clearly that this species belongs to the Tricharina gilva species complex and is different from T. ascophanoides in the sense of boudier. in contrast to this result, our re-examination of the collection rk 82.146 (teleomorph) revealed that it is morphologically highly similar to T. ascophanoides in the sense of boudier. in the light of this result, we also sequenced apothecia from rk 82.146. based on the lSu and iTS loci, this confirmed the result of our morphological study. The sequences of rk 82.146 and of cSY 123 7 — the anamorph culture of rk 82.146 — are different! 6 historical province, see https://en.wikipedia.org/wiki/Tavastia_(historical_province) 7 Sequenced by EGGEr (1996). 114 Ascomycete.org Tricharina ascophanoides is a rather common species in mountainous areas in central Europe (also at lower elevation in nordic countries), often collected in forest along paths at anthropogenic influenced areas. its determination is quite easy. illustrations of this species can be found in boudiEr (1907b, pl. 351), EnGEl & häffnEr (1986), douGoud (2002) and van voorEn (2014). it mainly differs from the other Tricharina species by the absence of true marginal hairs. only short hyphoid and hyaline hairs can be observed at the margin. on the outer surface, hyphoid hairs, typically with a bulbous base, are present. This type of hair is very similar to the type found on colored species of “Leucoscypha”, i.e. L. patavina or L. semi-immersa. recent phylogenetic analyses inferred that T. ascophanoides is affiliated with colored “Leucoscypha”, but in a sister clade together with Cupulina montana dougoud, van vooren & m. vega. Cupulina is a recently described monotypic genus with C. montana as type species (see douGoud et al., 2015). Since molecular phylogenetic analyses and morphological characters provide strong evidence that T. ascophanoides is closely allied with C. montana, we here exclude it from the genus Tricharina s. str. and transfer T. ascophanoides to Cupulina. Taxonomy Cupulina ascophanoides (boud.) van vooren, comb. nov. – mb 819489 basionym: Tricharia ascophanoides boud., Bull. Soc. mycol. Fr., 19 (3): 195 (1903). homotypic synonyms: Lachnea ascophanoides (boud.) Sacc. & d. Sacc., Syll. fung., 18: 37 (1905); Tricharina ascophanoides (boud.) chin S. Yang & korf, Mycotaxon, 23: 485 (1985). misapplication: Ascorhizoctonia ascophanoides chin S. Yang & r. kristiansen, Mycotaxon, 35 (2): 314 (1989), which represents a member of the genus Tricharina s. str. as circumscribed in this paper. Apothecia 1–4 mm diam., discoid or weakly cupulate, hymenium plan or a bit depressed, pale beige to ochraceous beige, or dull orange; external surface concolor, looking alike glabrous. Margin sometimes a bit eroded, without hairs. Ectal excipulum up to 500 µm thick, of textura globulosa/angularis, with cells 6–40 µm wide, intermixed with some piriform or cla- Plate 6 – Cupulina ascophanoides. a. coll. n.v. 2006.08.24. b. coll. n.v. 2007.07.30. c. coll. n.v. 2015.06.04. d. ascospores from coll. n.v. 2007.07.30, in water (Scale bar = 10 µm). E. figures of Tricharia ascophanoides adapted from boudier’s plate 351 in his Icones. f. coll. u.l. 131. Ascomycete.org 115 A B C Fig. 5 – Cupulina ascophanoides. micro-characters. a. ascospores, in water. b. Top of ascus and paraphyses. c. marginal cells and hyphoid hairs. Scale bars = 10 µm. a-b from coll. n.v. 2014.06.07, c from coll. n.v. 2015.06.14. 116 Ascomycete.org Table 3 – comparison between Cupulina montana and C. ascophanoides Cupulina montana Cupulina ascophanoides Macroscopic habit apothecia sessile, cupulate to hemispherical, sometimes apothecia sessile, hemispherical, rarely cupulate, Ø 1– plane, Ø (1) 2–4 mm, ochraceous yellow to orange 4 mm, ochraceous beige to orange beige Ascospore shape fusoid, often inequilateral Ellipsoid, sometimes a bit tapered at the poles Ascospore size 27–35 (36) × 12–13.6 (14) μm, Qm = 2.5 16.5–22.5 (24) × (9.5) 10–13 (14.5) μm, Qm = 1.7 Ascospore content filled with oil drops Without oil drops, but with bSG External surface anchor hyphae present, with a bulbous base, reaching 1000 μm in length hyphoid hairs present, with an enlarged or subbulbous base, reaching 450–500 μm in length Ecology in mountainous area, on calcareous soil, on naked or mossy ground in mountainous area, on naked ground Phenology from July to September from June to September vate hyphae. Medullary excipulum and subhymenium not distinct, rather thin, about 120–130 µm thick, of textura intricata, with hyaline or slightly pale orange hyphae. Marginal cells organized in a textura prismatica, the terminal cell often clavate, 10–11 µm wide, with some scattered, hyaline, hyphoid hairs emerging, 80–120 × 5– 8 µm, with a simple base, up to 0.5 µm thick-walled, apically obtuse or tapered. Excipular hairs present, hyphoid, 70–240 × 3–6 µm, hyaline, more or less flexuous, septate, with an enlarged to subbulbous base, up to 1 µm thick-walled, usually sharp at the top. Asci cylindrical, 160–220 × 14–21 µm, with crozier, 8-spored, operculate, inamyloid. Paraphyses not or only a few enlarged at the top, 3–5 (8) µm wide, septate, hyaline, without vacuoles but with some small inclusions, not reacting in mlZ. Ascospores uniseriate, ellipsoid, sometimes a bit tapered at the poles, (16) 16.5–22.5 (24) × (9.5) 10–13 (14.5) µm [X = 18.7 × 11.3 µm], Q = 1.3–1.9 [Qm = 1.7], hyaline, thinwalled or slightly thick-walled, without oil drops but containing bipolar spore granules, smooth, partially refractive in cb. Studied collections: francE – doubs, levier, route forestière de ravonnet, 46.929224° n 6.080289° E, alt. 750 m, leg. G. moyne, on soil, under Picea abies, 30 aug. 2006, pers. herb. n.v. 2006.08.24. doubs, levier, route forestière du Pont de la marine, 46.9480952° n 6.0671202° E, alt. 705 m, leg. n. van vooren, on soil, under Picea abies, 20 Jul. 2007, pers. herb. n.v. 2007.07.30. Jura, la Pesse, forêt de chapuzieux, 46.27027° n 5.827954° E, alt. 1185 m, leg. n. van vooren, on soil, in a mixed forest, 18 Sept. 2009, pers. herb. n.v. 2009.09.24. hautes-alpes, allos, digue d’allos, 44.2226786° n 6.6350181° E, alt. 1365 m, leg. n. van vooren, on soil, 6 Jun. 2010, pers. herb. n.v. 2010.06.37. Savoie, beaufort, la Tetraz, 45.707127° n 6.615065° E, alt. 1000 m, leg. n. van vooren, on soil, 28 aug. 2013, pers. herb. n.v. 2013.08.53. isère, lans-en-vercors, bois de chabaud, route forestière de Servagnet, 45.1159° n 5.5488° E, alt. 1258 m, leg. n. van vooren, on soil, 26 Sept. 2013, pers. herb. n.v. 2013.09.31. Savoie, bonnevalsur-arc, Pont de la neige, 45.40652° n 7.02843° E, alt. 2520 m, leg. n. van vooren, on soil, in alpine grassland, 24 aug. 2015, pers. herb. n.v. 2015.08.15. SWiTZErland – fribourg, bulle, cabane d’inson, 46.584153° n 7.0010782° E, alt. 1025 m, leg. n. van vooren, on soil, 23 Jun. 2012, pers. herb. n.v. 2012.06.20. Idem, 20 Jun. 2014, pers. herb. n.v. 2014.06.07. Idem, 5 Jun. 2015, pers. herb. n.v. 2015.06.04. auSTria – carinthia, Weißensee (ostufer), 46.704189° n 13.413577° E, alt. 940 m, leg. and det. b. Wergen, at a wayside on soil between mosses within a Picea-forest, 8 Jun. 2013, pers. herb. u.l. 85. iTalY – South Tyrol, Sesto, 46.685679° n 12.375881° E, alt. 1420 m, leg. and det. u. lindemann, at a wayside on soil, 16 aug. 2009, pers. herb. u.l. 33. Slovakia – vysoké Tatry, Siedmich valley, 49.21651° n 20.290085° E, alt. 1010 m, leg. and det. u. lindemann , on soil, 10 Sept. 2012, pers. herb. u.l. 131. vysoké Tatry, near Ždiar, 49.26941° n 20.270406° E, alt. 880 m, leg. and det. u. lindemann, on soil at the edge of a burnt site, 14 Sept. 2012 (only macro- and micropictures, no exsiccate). norWaY – Østfold county, fredrikstad community, borge, Torp, 59.240630° n 11.007230° E, alt. 13 m, leg. m. Pettersen, det. r. kristiansen, rev. u. lindemann, closed paper mill area, on artificial calcareous ground with few mosses, created by man, 2 Jun. 2014, pers. herb. u.l. 192. Idem, leg. and det. r. kristiansen, rev. u. lindemann and n. van vooren, on balls of cellulose, partially burnt, 3 Jul. 1982, pers. herb. r.k. 82.146. Distribution: north and central Europe – austria (our data), czech republic (YanG & korf, 1985b), france (boudiEr, 1903; our data), Germany (EnGEl & häffnEr, 1986), italy (our data), norway (kriSTianSEn, 2014, pro parte 8; our data), Slovakia (YanG & korf, 1985b; our data), and Switzerland (douGoud, 2002; our data). Comments: although Cupulina montana and C. ascophanoides are in some aspects morphologically similar, the taxonomic position of the latter was a bit surprising due to the different ascospore structure of each species. Table 3 gives a comparison of them. Two attempts to sequence the holotype of Tricharia ascophanoides failed due to a fungal contamination, but we think that boudier’s protologue, our re-examination of type-material and the characters consistent with the species description for T. ascophanoides coming from the numerous collections we studied are sufficient to circumscribe this species without ambiguity. The Sepultariella clade PErrY et al. (2007) showed that some tricharinoid species attached to Pustularia (as “P. patavina”) or Leucoscypha (as L. semiimmersa) do not belong to either of these genera. our own analyses confirm that. Pustularia patavina cannot be retained because the name Pustularia is illegitimate (icn art. 53.1), and L. semi-immersa does not belong to the same clade as the type species of Leucoscypha, L. leucotricha (alb. & Schwein.) boud. They were included in Leucoscypha by SvrčEk (1974), following the enlarged concept of this genus proposed by Saccardo (1889), lE Gal (1957), rifai (1968), Eckblad (1968), PanT & TEWari (1977) and many others. Thus Leucoscypha became a heterogeneous assemblage of species, not supported by phylogenetic analyses. kuTorGa (2000) used the name Sepultariella for this group but unfortunately the publication of this name is invalid (icn art. 39.1). as it is registered in well-known fungal nomenclature databases (index fungorum and mycobank), we think that it is better to validate Sepultariella than to create another name. We therefore propose the newly validated genus Sepultariella to accommodate P. patavina and L. semi-immersa, the first two species included in this genus. We choose Peziza patavina as type species because S. semiimmersa could represent a species complex. 8 The collection r.k. 82.146 that we re-examined corresponds to C. ascophanoides, but the fungus illustrated by kriSTianSEn (2014: 50) does not look like the latter and is probably another Tricharina species. Ascomycete.org 117 Sepultariella van vooren, u. lindemann & healy, gen. nov. – mb 819861 diagnosis: differs from Leucoscypha boud. by pigmented apothecia and smooth ascospores. Type species: Peziza patavina cooke & Sacc. Etymology: from latin sepultus that means buried. Sepultariella patavina (cooke & Sacc.) van vooren, u. lindemann & healy, comb. nov. – mb 819862 basionym: Peziza patavina cooke & Sacc., in Saccardo, Michelia, 1: 70 (1877). homotypic synonyms: Neottiella patavina (cooke & Sacc.) Sacc., Syll. fung., 8: 193 (1889); Humaria patavina (cooke & Sacc.) rehm, Rabenh. Kryptog.-Fl., 1 (3): 957 (1894); Pustularia patavina (cooke & Sacc.) boud., Icon. mycol., liste prélim.: 3 (1904); Leucoscypha patavina (cooke) Svrček, Česká Mykol., 28 (3): 132 (1974); Leucoscypha patavina (cooke) Pant & v.P. Tewari, Trans. Brit. Mycol. Soc., 68 (3): 441 (1977), illeg. Sepultariella semi-immersa (P. karst.) van vooren, u. lindemann & healy, comb. nov. – mb 819863 basionym: Peziza semi-immersa P. karst., Notis. Sällsk. Fauna Fl. Fenn. Förhandl., 10: 117 (1869). homotypic synonyms: Humaria semi-immersa (P. karst.) Sacc., Syll. fung., 8: 143 (1889); Sepultaria semi-immersa (P. karst.) massee, Brit. Fung.-Fl., 4: 391 (1895); Humarina semi-immersa (P. karst.) Seaver, N. Amer. Cup-Fungi (Operc.): 130 (1928); Leucoscypha semi-immersa (P. karst.) Svrček, Česká Mykol., 28 (3): 133 (1974); Octospora semi-immersa (P. karst.) k.b. khare & v.P. Tewari, Canad. J. Bot., 56 (17): 2118 (1978); Byssonectria semi-immersa (P. karst.) benkert, Gleditschia, 15 (1): 181 (1987). Outlook here we focused on the four key species of Tricharina and the delimitation of the most closely allied genera. in an upcoming manuscript, we will focus on the other species of the genus which are until now not sufficently known in regard to their taxonomic position, that include: T. flava, T. groenlandica, T. herinkii, T. hiemalis, T. japonica, T. pallidisetosa, T. striispora, and T. tophiseda. These species will be also analyzed morphologically and phylogenetically based on type studies and recent findings. on the basis of the results, a key will be provided with which it will be possible to distinguish the morphologically often very similar species of Tricharina and those of closely allied genera. Acknowledgements We acknowledge gratefully bernard clesse, rené dougoud, Zuzana Egertová, bernd fellmann, felix hampe, Jesko kleine, roy kristiansen, masanori kutsuna, rubén martinéz Gil, Gilbert moyne, alija mujic, rubén negrín, Peter Püwert, miguel ángel ribes, Torsten richter, rika Seibert, beatrice Senn-irlet, matthew Smith, Salvador Tello and marcel vega for having shared their collections, photographs or data. The curators of the following herbaria are acknowledged for their loans: bart buyck (Pc, muséum national d’histoire naturelle, france), begoña aguirre-hudson (k, royal botanic Gardens kew, united kingdom), Genevieve E. Tocci (fh, farlow herbarium, harvard university, uSa), anna-lena anderberg and Jens klackenberg (S, naturhistoriska riksmuseet, Sweden). 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WhiTnEY h.S. & ParmETEr J.r. 1964. — The perfect stage of Rhizoctonia hiemalis. Mycologia, 56 (1): 114-118. YanG c.S. & korf r.P. 1985a. — Ascorhizoctonia gen. nov. and Complexipes emend., two genera of anamorphs of species assigned to Tricharina (discomycetes). Mycotaxon, 23: 457-481. YanG c.S. & korf r.P. 1985b. — a monograph of the genus Tricharina and a new, segregate genus, Wilcoxina (Pezizales). Mycotaxon, 24: 467-531. YanG c.S. & kriSTianSEn r. 1989. — Ascorhizoctonia ascophanoides sp. nov.: anamorph of Tricharina ascophanoides. Mycotaxon, 35 (2): 313-316. Appendix This appendix contains the list of sequences used during our study to release the phylotrees (fig. 1 and 2). Ascomycete.org 119 120 Ascomycete.org Original name (identified as…) Correct name (if different) Cupulina montana Cupulina montana GenBank Number Voucher or Strain Number Country Collector(s) ITS LSU n.v. 2011.09.04 france n. van vooren kY364048 kY364070 Switzerland r. dougoud kY364047 kY364073 Tricharina gilva complex (hh) 83/168 Switzerland b. Senn-irlet kY928062 kY928059 Leucoscypha patavina Sepultariella patavina n.v. 2011.09.18 france n. van vooren kY364045 kY364075 Leucoscypha patavina Sepultariella patavina n.v. 2013.08.71 Georgia Z. Egertova kY364042 kY364076 Leucoscypha patavina Sepultariella patavina r.d. 15.50.227.91 Switzerland r. dougoud kY364043 kY364077 Leucoscypha patavina Sepultariella patavina m.v. 20131018-4 Germany m. vega kY364044 kY364078 Leucoscypha cf. patavina Sepultariella patavina n.v. 2013.11.17 Portugal n. van vooren kY364037 kY364081 Leucoscypha semi-immersa Sepultariella semi-immersa r.d. 25.09.227.98 Switzerland r. dougoud kY364036 kY364079 Leucoscypha semi-immersa Sepultariella semi-immersa nv 2006.09.27 france n. van vooren kY364040 kY364074 Leucoscypha semi-immersa Sepultariella semi-immersa m.v. 20130928-3 Germany m. vega kY364038 kY364082 Leucoscypha semi-immersa Sepultariella semi-immersa m.v. 20131025-2 Germany m. vega kY364039 kY364083 Paratricharina cf. poiraultii Paratricharina poiraultii u.l. 116 Spain f. hampe & J. kleine kY364029 kY364053 Pseudaleuria fibrillosa u.l. 182 Germany m. vega kY364035 kY364068 Pseudotricharina sp. mES-2152 argentina r. healy kY364033 kY364072 mES-2065 argentina d. Torres acuna kY364034 kY364071 u.l. 181 Ethiopia u. lindemann kY364041 kY364080 Pseudotricharina sp. Pustularia cf. patavina Sepultariella semi-immersa complex Tricharina aethiopica mSTr P-19994 (holotype) Ethiopia u. lindemann kY364022 kY364061 Tricharina ascophanoides Cupulina ascophanoides n.v. 2014.06.07 Switzerland n. van vooren kY364046 kY364069 Tricharina ascophanoides Cupulina ascophanoides n.v. 2006.08.24 france n. van vooren kY364049 kY364051 Tricharina ascophanoides Cupulina ascophanoides n.v. 2007.07.30 france n. van vooren kY364050 kY364052 Tricharina ascophanoides Cupulina ascophanoides u.l. 131 Slovakia u. lindemann kY928063 kY928060 Tricharina ascophanoides Cupulina ascophanoides u.l. 192 norway m. Pettersen kY928064 kY928061 Tricharia gilva Tricharina gilva Pc 0167813 (lectotype) france E. boudier kY364019 kY364057 Tricharina cf. gilva Tricharina hiemalis n.v. 2011.08.14 france n. van vooren kY364021 kY364059 Tricharina cf. gilva Ascorhizoctonia praecox n.v. 2016.03.01 france b. clesse kY364031 kY364054 Tricharina cf. gilva r.m.-1254 Spain r. martinéz Gil kY364028 kY364063 Tricharina glabra mSTr P-19995 (holotype) Germany T. böhning kY364024 kY364067 Tricharina ochroleuca Tricharina gilva complex n.v. 2011.08.05 france n. van vooren kY364027 kY364060 Tricharina ochroleuca Tricharina gilva u.l. 191 Germany r. Seibert kY364020 kY364065 Tricharina ochroleuca Tricharina gilva complex u.l. 241 czech republic Z. Egertova - kY364066 Tricharina ochroleuca Tricharina gilva complex u.l. 243 Spain r. martinéz Gil kY364026 kY364064 Tricharina cf. ochroleuca Tricharina gilva complex n.v. 2013.06.00 Spain G. moyne kY364025 kY364058 Tricharina praecox Ascorhizoctonia praecox u.l. 100 Spain r. negrín kY364030 kY364055 Tricharina praecox var. cretea Ascorhizoctonia praecox r.d. 15.06.245.91 Switzerland r. dougoud kY364032 kY364056 Ja-cuSSTa: 7799 Spain S. Tello kY364023 kY364062 Tricharina striispora Table 4 – list of collections of Tricharina and other genera sequenced during this study G 00262366 (holotype) Geopora herinkii Original name (identified as…) Correct name (if different) Anthracobia macrocystis Anthracobia sp. Ascorhizoctonia ascophanoides Tricharina gilva complex Ascorhizoctonia sp. (culture) Tricharina sp. Country Collector(s) c kh.01.035 denmark k. hansen QcnE Tl-11709 Ecuador cSY 123 (holotype) norway r. kristiansen cbS 240.85 (holotype) norway h. dissing & S. Sivertsen GenBank Number ITS kc012664 u38581 u38580 JQ836559 JQ836562 JQ836563 bcS-1 china ? Z.l. Yuan JQ836557 Ascorhizoctonia sp. (endophyte) 23 hkd-2015 iran h.k. dolatabad et al. kP641147 Ascorhizoctonia sp. (endophyte) cSE195 iran J. Soltani et al. kJ598625 Ascorhizoctonia sp. (endophyte) cSE148 iran J. Soltani & S. Pakvaz kT024986 Ascorhizoctonia sp. (endophyte) Tricharina sp. Zc-W-2-2 china f. bu & Z. Yu kr822155 Ascorhizoctonia sp. (endophyte) Tricharina gilva complex 23 hkd-2015 iran h.k. dolatabad et al. kP641147 fungal sp. (endophyte) Tricharina gilva complex bG46 france a. langenfeld et al. fungal sp. (orchid mycorrhiza) Tricharina sp. cc26 argentina S. fracchia et al. Geopora arenicola c kS-94-173 denmark k. hansen & S.k. Sandal Geopora cercocarpi kropp2 uSa d. Southworth & J.l. frank Geopora cervina (Ecm) uu_3a_i Sweden JQ249225 kf151200 dQ220336 hQ283098 JQ724065 Geopora cf. cervina fh kh.03.61 norway Geopora clausa ah39177 Spain Jn812052 Geopora clausa ah39181 Spain Jn812053 Geopora clausa ma 46927 Portugal Geopora cooperi SfSu hdT 52489 uSa Geopora cooperi ah9065 Spain Geopora cooperi vk1783 Greece Geopora cooperi fh baP 517 uSa Geopora foliacea ah38936 Spain LSU kc012663 k. hansen dQ220344 JX424576 J. ammarati dQ220341 Jn812044 v. kaounas Jn812045 kc012678 Jn812047 Geopora foliacea ah38937 Spain Geopora nicaensis mcvE 14596 italy E. campo Jf908022 Geopora sumneriana mcvE 16978 italy a. montecchi Jf908024 Hoffmannoscypha pellita fh dhP 297 uSa d.h. Pfister Hoffmannoscypha pellita m Gh20100409 Germany d. Strobelt & G. hensel hQ913564 hQ993571 Lotinia verna SESTao 2003040401 (holotype) Spain J.l. Pérez-butrón kP195730 kP195729 Melastiza contorta c kh.01.06 Sweden k. hansen Paratricharina poiraultii u.l. 179-14 Spain T. illescas et al. kP052784 kP052785 u.l. 178-14 Portugal m. vega kP052788 kP052789 c14 china l.W. Shang Jn198476 Paratricharina poiraultii Pezizaceae sp. (endophyte) Tricharina gilva complex Jn812048 dQ220343 aY500539 Ascomycete.org Pezizomycetes sp. (endophyte) Tricharina gilva complex ariZ aEa dc2145 uSa b. klein et al. GQ153018 GQ153018 Pezizomycetes sp. (endophyte) Ascorhizoctonia praecox ariZ aEa 11056 uSa m. hoffman GQ153065 GQ153065 Pezizomycetes sp. (endophyte) Tricharina glabra mJ34 china f. huang et al. hQ449982 Pezizomycetes sp. (endophyte) Tricharina gilva complex dc2145 uSa b. klein et al. GQ153018 Table 5 – list of other sequences from the inSdc public databases also used in phylogenetic analyses Ascorhizoctonia praecox Voucher or Strain number 121 122 Ascomycete.org Original name (identified as…) Correct name (if different) GenBank Number Voucher or Strain Number Country Collector(s) Phaeangium lefebvrei ira-mba sb11 Tunisia i. Sbissi et al. Picoa juniperi ira-mba sb6 Tunisia i. Sbissi et al. Pseudaleuria quinaultiana oSc 45766 (paratype) uSa d. lusk-newman Pseudaleuria quinaultiana nSW 7107 uSa n.S. Weber Pseudotricharina intermedia ST 25011493 (holotype) Spain S. Tello kT861358 Pseudotricharina intermedia Gk 6904 Greece G. konstantinidis kT861359 Pustularia patavina Sepultariella patavina fh kh.03.73 norway k. hansen & c. lange ITS LSU Gu391557 Gu391558 Eu669387 dQ220389 kT861360 kT861361 dQ220396 Pyronemataecae sp. Cupulina ascophanoides fh kh.04.21 uSa n. Weber & k. hansen dQ220399 Sphaerosporella brunnea Trichophaea brunnea fh dhP dr.02.16 dominican republic d.h. Pfister dQ220432 Sphaerosporella brunnea Trichophaea brunnea fh kh.03.04 uSa k. hansen dQ220433 c f-55022 norway h. dissing dQ220431 dQ220442 Sphaerosporella hinnulea Tricharina gilva Ascorhizoctonia praecox c f-55212 norway h. dissing Tricharina gilva Ascorhizoctonia praecox dEd 7356 uSa d.E. desjardin dQ220443 Tricharina gilva Ascorhizoctonia praecox fh baP 431 uSa b.a. Perry dQ220444 Tricharina gilva not a Tricharina species uSa f.a. Wolf JQ824118 cbS 237.85 (holotype) Greenland h. dissing JQ824125 Tricharina hiemalis (culture as Rhizoctonia hiemalis) cbS 263.60 canada h.k. Saksena JQ824124 Tricharina ochroleuca Tricharina hiemalis (?) c f-53062 Greenland h. dissing Tricharina ochroleuca Tricharina groenlandica cbS 238.85 (neotype) norway S. Silvertsen & h. dissing Tricharina ochroleuca (Ecm? Endophyte?) u38576 dQ220445 JQ824126 JQ836561 ok3f101hP china X. Yang et al. kf871459 Tricharina praecox Tricharina gilva fh kh.03.101 norway k. hansen dQ646552 Tricharina praecox Ascorhizoctonia praecox m 0178315 Germany T. kupka JQ824122 Tricharina praecox Ascorhizoctonia praecox m 0178316 austria m. moser JQ824120 Tricharina sp. not a Tricharina species barr 5907 uSa m.E. barr Tricharina sp. c Tl-10051 Ecuador J. Salazar & T. læssøe Tricharina striispora (culture) bccm/mucl 41297 argentina Z.l. Yuan JQ836556 oT2-144 Portugal T. Gomes et al. kT804076 Trichophaea abundans cbS 348.76 finland v. hintikka dQ220448 Trichophaea hemisphaerioides fh kh.04.54 uSa k. hansen kc012710 Tricharina striispora (endophyte) Trichophaea hybrida Tricharina striispora Tricharina hiemalis (?) amnh-49682 iceland Trichophaea minuta cbS 236.57 (isotype) canada r.f. cain & n.a. hastings Trichophaea saccata cbS 804.70 (type) uk h.c. Evans Trichophaea woolhopeia n.v. 2006.10.01 france n. van vooren Trichophaea woolhopeia kh.01.033 denmark k. hansen dQ646525 dQ220446 dQ220447 JQ836560 dQ220455 dQ220452 dQ220451 kX982244 kX982243 dQ220460 uncultured Ascorhizoctonia clone (soil sample, ginger rhizosphere) bdr3c10 india h. Thakur et al. kJ591759 uncultured Geopora (orchid mycorrhiza) Geopora sp. lP34-Geo1 russia T. Tesitelova et al. kJ188583 uncultured Pezizaceae (orchid mycorrhiza) Pseudotricharina sp. 1221a South africa r.J. Waterman et al. fJ788784 Table 5 – (continued) nY nYbG 1968 Tricharina groenlandica Table 5 – (continued) Original name (identified as…) Correct name (if different) Voucher or Country Strain Number uncultured Pezizaceae (orchid mycorPseudotricharina sp. rhiza) uncultured Pezizaceae (orchid mycorPseudotricharina sp. rhiza) uncultured Pyronemataceae (orchid myTricharina hiemalis chorrhiza) Wilcoxina mikolae Collector(s) GenBank Number ITS 1223a South africa r.J. Waterman et al. fJ788780 1186a South africa r.J. Waterman et al. fJ788782 3075 Germany m.i. bidartondo aY634115 SfSu WS 36 uSa W. Stoll Wilcoxina mikolae hmJau27330 china c.h. Shi & T. bau ku061020 Wilcoxina rehmii (Ecm) nS211 lithuania a. menkis et al. dQ069001 LSU dQ220468 ef Nicolas Van Vooren 36 rue de la Garde 69005 lyon france nicolas@vanvooren.info Uwe Lindemann Pflügerstr. 62 12047 berlin Germany uwe.lindemann0907@gmail.com Rosanne Healy department of Plant Pathology university of florida, Gainesville, fl 32611 uSa rhealy1@ufl.edu Ascomycete.org 123