Ceratolenta caudata gen. et sp. nov. and Platytrachelon abietis gen. et ...

Mycologia, 105(2), 2013, pp. 462–475. DOI: 10.3852/12-199 # 2013 by The Mycological Society of America, Lawrence, KS 66044-8897

Two taxonomic novelties in the Sordariomycetidae: Ceratolenta caudata gen. et sp. nov. and Platytrachelon abietis gen. et comb. nov. for Ceratosphaeria abietis Martina Re´blova´1

simple and indistinct morphology of dark ascomata with carbonized walls, long necks or beaks and hyaline to subhyaline, ellipsoidal, fusiform to cylindrical ascospores in cylindrical-clavate stipitate asci. Among them are several small or monotypic genera, which are distantly related in this fungal subclass, some of them placed in an incertae sedis position. Among the first generic names that match these characteristics, Ceratosphaeria Niessl (Niessl 1876), Ceratostomella Sacc. (Saccardo 1878) and Lentomitella Ho¨hn. (von Ho¨hnel 1905) have been recognized for more than a century to harbor morphologically similar but phylogenetically unrelated organisms. In recent years DNA sequence data have helped to clarify their phylogenetic relationships. Based on molecular analyses of small and large subunit nuclear ribosomal DNA (nc18S and nc28S), the three genera have been reinstated and their descriptions emended (Hyde et al. 1997, Re´blova´ 2006, Huhndorf et al. 2008). In our phylogenetic analyses they always cluster in a robust subclade of the Sordariomycetidae, which includes the Ophiostomatales and four other families. They form three strongly supported monophyletic evolutionary lineages. Ceratosphaeria and the two segregate genera Ceratosphaerella Huhndorf et al. and Muraeriata Huhndorf et al. are accommodated in the Magnaporthaceae, while Ceratostomella and Lentomitella remain in the incertae sedis position. In addition, Ceratostomella deviates morphologically slightly from Ceratosphaeria and Lentomitella. It differs by pigmented ascospores in smaller asci with an indistinct apical annulus and presence of characteristic ascogenous hyphae with croziers, each producing lateral and terminal dehiscent cells from each of which one ascus arises as an outgrowth (Re´blova´ 2006). Molecular data have helped to recognize additional morphologically similar taxonomic novelties in the Sordariomycetidae, including freshwater and terrestrial taxa (e.g. Ho et al. 1999, Hyde 1992, Wong and Hyde 1999, Wong et al. 1999, Raja et al. 2003, Tsui et al. 2003, Shearer et al. 2003, Campbell and Shearer 2004, Arzanlou et al. 2007, Re´blova´ and Sˇteˇpańek 2009, Re´blova´ 2009, Thongkantha et al. 2009, Wong et al. 2009, Abdel-Wahab et al. 2011, Ferrer et al. 2012, Liu et al. 2012). Recent collections of an undescribed wood-inhabiting fungus and of Ceratosphaeria abietis Re´blova´ (Re´blova´ 1998) reveal two additional evolutionary

Department of Taxonomy, Institute of Botany of the Academy of Sciences, CZ-252 43, Pru˚honice, Czech Republic

Abstract: Wood-inhabiting taxa of the Sordariomycetidae comprise several distantly related monotypic or small genera, which lack familial or ordinal affiliation and share a simple and inconspicuous morphology of dark ascomata with carbonaceous walls and long necks, stipitate asci and hyaline ellipsoidal, fusiform to cylindrical ascospores. Recent collections of an undescribed fungus and of Ceratosphaeria abietis reveal two additional evolutionary lineages characterized by this simple and indistinct teleomorph morphology. Phylogenetic analysis of three genes, small and large subunit nuclear ribosomal DNA (nc28S and nc18S rDNA) combined with the second largest subunit of RNA polymerase II (rpb2), supports the recognition of two new genera, Ceratolenta and Platytrachelon for C. abietis. Platytrachelon abietis is redescribed and illustrated based on additional collections. In culture it produced a dematiaceous hyphomycetous anamorph with blastic conidiogenesis and ellipsoidal, septate, pale brown conidia. It was associated with a synanamorph producing cylindrical, strongly curved hyaline conidia. Molecular data suggest a relationship of Platytrachelon with the Papulosaceae, while Ceratolenta forms a monophylum on a separate branch. Both taxonomic novelties possess striking morphological similarities with Ceratosphaeria, Lentomitella and Rhodoveronaea, which recently were reinstated based on DNA sequence data. A key to morphologically similar wood-inhabiting fungi classified in the Sordariomycetidae is provided. Key words: Annulatascaceae, Lentomitella, life history, Magnaporthaceae, systematics, three-gene phylogeny INTRODUCTION Wood-inhabiting ascomycetes classified in the Sordariomycetidae comprise many genera, which share a Submitted 4 Jun 2012; accepted for publication 4 Sep 2012. 1 Corresponding author. E-mail: [email protected]

462

RE´BLOVA´: TWO TAXONOMIC NOVELTIES IN SORDARIOMYCETIDAE lineages characterized by a simple and inconspicuous teleomorph morphology, which makes them difficult to identify. An undescribed perithecial ascomycete was collected frequently on decaying deciduous wood in mountainous regions in the Czech Republic. The teleomorph possesses dark brown immersed perithecia with long emerging necks, asci with a conspicuous apical annulus and long tapering stipes and containing eight hyaline, fusiform, septate ascospores. No conidiophores were observed on the natural substratum, and the culture derived from isolated ascospores remained sterile. Among comparatively well known ascomycetes, the fungus is morphologically remarkably similar to Fluminicola G.S. Wong et al. (Wong et al. 2009) and Rhodoveronaea Arzanlou et al. (Arzanlou et al. 2007, Re´blova´ 2009). A culture derived from a recent collection of Ceratosphaeria abietis yielded a dematiaceous hyphomycete with blastic conidiogenesis and spherical subhyaline conidiogenous cells producing ellipsoidal, one-septate, pale brown conidia. A synanamorph was observed in vitro, forming scarcely growing phialides producing cylindrical, strongly curved hyaline conidia. Comparison of the type and two other collections made in the same locality in the Czech Republic with specimens from different localities and country indicates that the three former specimens exhibited a certain abnormality in ascospore development, which is less prominent or lacking in other collections. Therefore, the species description of C. abietis is emended based on new observations and cultivation studies. The anamorph is morphologically similar to the anamorph genus Triadelphia Shearer & Crane, whose species are characterized by up to eight kinds of conidia developed during ontogeny. This study discloses and clarifies the phylogenetic relationships of C. abietis and the unknown ascomycete with Ceratosphaeria of the Magnaporthaceae and other morphologically similar fungi of the Sordariomycetidae. We used sequence data from members of the Sordariomycetes, two functional ribosomal RNA genes, the small and large subunit of the nuclear ribosomal RNA (nc18S and nc28S), and a proteincoding gene, the second largest subunit of RNA polymerase II (rpb2). MATERIALS AND METHODS Morphological observations of herbarium materials and fungal strains.— Herbarium specimens examined and cultures studied are listed under the species treated. Dry stromata were rehydrated with water; material was examined with an Olympus SZX12 dissecting microscope and centrum material (including asci, ascospores and paraphyses) was mounted in Melzer’s reagent or 90% lactic acid. Hand sections of the perithecium wall were studied. When

463

present, conidiophores, conidiogenous cells and conidia were examined as microscope slide preparations mounted in water or Melzer’s reagent. All measurements were made in Melzer’s reagent. Means 6 standard errors (SE) based on 20–25 measurements are given for dimensions of asci, ascospores, conidia and conidiogenous cells. Images were captured with differential interference (DIC) or phase contrast (PC) microscopy with an Olympus DP70 Camera operated by Imaging Software Cell on an Olympus BX51 compound microscope. Multi-ascospore isolates were obtained from fresh material with a spore isolator (Meopta, Prague, Czech Republic). Isolates were grown on potato-carrot agar (PCA) and 2% malt extract agar (MEA; malt extract, Oxoid) (Gams et al. 1998). Colonies were examined after 7, 21 and 30 d at 25 C in the dark. Cultures are maintained at CBS (CBS-KNAW Fungal Biodiversity Center, Utrecht, the Netherlands). Type and other important herbarium material are deposited in PRM herbarium (National Museum Prague). Abbreviations of collectors on specimen labels refer to Martina Re´blova´ (M.R.) and Jacques Fournier (J.F.). DNA extraction, amplification and sequencing.— These procedures were performed as described in Re´blova´ et al. (2011). DNA extraction from dried ascomata (PRM 899855) followed the same procedure as extraction from living cultures, with a variation in the first step of the protocol. Ascomata were rehydrated in 350 mL mixture of MicroBead and M1 solutions (UltraClean Microbial DNA Kit, MoBio Laboratories Inc., Canada), followed by freezing overnight at 220 C. Sequence alignment.—Accession numbers and isolate information for new nc28S, nc18S and rpb2 sequences derived from DNA extracts of multiple-ascospore isolates of Ceratosphaeria abietis, the undescribed ascomycete and other newly sequenced taxa are provided (TABLE I). The new sequences were aligned with homologous sequences retrieved from GenBank. The selection of retrieved sequences was adjusted according to the BLAST top hits for each gene. The GenBank accession numbers of the sequences selected for phylogenetic analyses are given in the tree after the taxon names (FIG. 1). All sequences were manually aligned in BioEdit 7.0.9.0 (Hall 1999). All nc18S and nc28S alignments were enhanced with the homologous 2D structure of Saccharomyces cerevisiae Meyen ex E.C. Hansen (Gutell 1993, Gutell et al. 1993) to improve the decisions on homologous characters and introduction of gaps. The alignment of rpb2 sequences was transformed into a protein alignment, maintaining a correct reading frame, with BioEdit. We applied Clustal W implemented in BioEdit and protein weight matrix Blosum62 to create the multiple sequence alignment. This alignment was corrected manually and further improved by considering the exchangeability of amino acids with similar chemical properties at certain positions. The protein alignment then was converted back into a DNA alignment. The multiple sequence alignment (nc18S-nc28S-rpb2) was constructed. To combine sequences for a multigene alignment, the individual alignments were concatenated

464 TABLE I.

MYCOLOGIA Sources and accession numbers of isolates and collections sequenced for this study GenBank accession nos. a

Source

Host/substrate

Locality

nc28S

nc18S

rpb2

Ceratolenta caudata

CBS 125234

Czech Republic

JX066704

JX066708

JX066699

Ceratolenta caudata

PRM 899855

Decaying deciduous wood Decaying wood of Acer pseudoplatanus Bark of Abies alba Submerged wood of Alnus glutinosa Decaying deciduous wood Decaying wood of Carpinus betulus Decaying deciduous wood

Czech Republic

JX066705

JX066709

–

Czech Republic France

JX066703 JX066706

JX066707 –

JX066698 –

Czech Republic

FJ617561b

JX066711

JX066702

Czech Republic

FJ617559b

–

JX066701

Sweden

FJ617560b

JX066710

JX066700

Taxon

Platytrachelon abietis CBS 125235 Pseudohalonectria lutea CBS 126574 Rhamphoria delicatula

CBS 132724

Rhodoveronaea varioseptata Rhodoveronaea varioseptata

CBS 123472 CBS 123473

a

CBS 5 CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands. PRM 5 Mycological Herbarium, National Museum Prague, Czech Republic. b Published sequences (Re´blova´ 2009). into one. The given lengths of the alignment were assessed after introduction of gaps (see RESULTS). The alignment is deposited in TreeBASE (Study no. 12739). Phylogenetic analyses.— Phylogenetic relationships were examined with nc28S, nc18S rDNA and rpb2 sequences from 91 taxa (88 species) from 18 orders or families of the Sordariomycetes. We analysed the first two-thirds of the 59 half of the nc28S (Gutell et al. 1993; D1 and D2 domains corresponding to the first 1197 nucleotides of Saccharomyces cerevisiae), the entire nc18S, and the 5–7 segment of the rpb2. Bases 1–75 were excluded from phylogenetic analyses of the nc28S and nc18S alignments and bases 1–60 were excluded from the analysis of rpb2 alignment because of the incompleteness of the 59-end of the majority of the available sequences. Two outgroup taxa, S. cerevisiae and Vanderwaltozyma polyspora (van der Walt) Kurtzman (Saccharomycetes), were used to root the phylogeny. The combined dataset was partitioned into four subsets of nucleotide sites: (i) nc28S, (ii) nc18S genes, (iii) first and second codon positions of rpb2, (iv) third codon position of rpb2. The three genes in the combined analysis were tested for heterogeneity between data partitions before combining them in a total evidence analysis. We used the partition homogeneity/incongruence-length difference test implemented in PAUP (Swofford 2002) to determine whether different partitions of the data have significantly different signals. Because combining data with value P , 0.01 generally improves phylogenetic accuracy (Cunningham 1997) and our data did not show significant heterogeneity (P 5 0.01), the sequences were combined for the further analyses. Maximum likelihood analysis (ML) was performed with RAxML-HPC 7.0.3 (Stamatakis 2006) with a GTRCAT model of evolution, which is a combination of GTRGAMMA and GTRCAT (a RAxML-specific alternative model, in which the alignment sites are pooled into a prespecified number of rate categories). The GTRCAT model is used for the heuristic

search and the best tree then is optimized and the likelihood values are calculated under the GTRGAMMA model. The nodal support was verified by nonparametric bootstrapping (BS) with 1000 replicates. Bayesian analysis (BAY) was performed in a likelihood framework as implemented by MrBayes 3.0b4 software package to reconstruct phylogenetic trees (Huelsenbeck and Ronquist 2001). MrModeltest2 2.3 (Nylander 2008) was used to infer the appropriate substitution model, which would best fit the model of DNA evolution for our sequence datasets. Multiple Bayesian searches using metropoliscoupled Markov chain Monte Carlo sampling were conducted. One cold and three heated Markov chains were used in the analysis. Bayesian analysis was run 5 000 000 generations, with trees sampled every 1000 generations. The first 20 000 trees, which represented the burn-in phase of the analysis, were discarded. To estimate posterior probabilities (PP) of recovered branches (Larget and Simon 1999) 50% majority rule consensus trees were created from the remaining trees with PAUP.

RESULTS The combined alignment of all three genes consisted of 4297 total characters (after introduction of gaps). The ML tree is illustrated (FIG. 1). For the BAY analysis, the GTR + I + GAMMA substitution model was inferred for each partition. In the ML and BAY analyses, the Sordariomycetidae (99% ML BS and 0.98 PP) contain three major clades, clade I (95/1.0) including the Boliniales, Coniochaetales, Chaetosphaeriales and Sordariales, clade II (94/1.0) including the Calosphaeriales, Diaporthales and Togniniaceae, and a robust clade III (94/1.0). The latter clade encompasses two subclades of fungi, many of which

RE´BLOVA´: TWO TAXONOMIC NOVELTIES IN SORDARIOMYCETIDAE

465

FIG. 1. Phylogenetic analysis of the Sordariomycetes including representatives of the three subclasses, Hypocreomycetidae, Sordariomycetidae and Xylariomycetidae. Phylogram inferred from combined nc18S-nc28S-rpb2 sequences with maximum likelihood analysis using a GTRCAT model of evolution. Maximum likelihood bootstrap support (ML BS) and Bayesian posterior probability (PP) are indicated at the nodes. Branches with a black oval symbol indicate ML BS 5 100%, PP values 5 1.0. The GenBank accession numbers given in the tree after the names are those of nc28S/nc18S/rpb2 genes. Missing sequences are indicated by –.

466

MYCOLOGIA

share the simple and inconspicuous morphology of hyaline ascospores, stipitate asci and dark ascomata with long necks or beaks. These are the Magnaporthaceae (94/1.0) and the second more heterogeneous subclade (66/1.0) containing the Ophiostomatales, three other families, namely the Annulatascaceae, Papulosaceae and Thyridiaceae, and several other monophyletic genera without family or order affiliation. Based on results of the three-gene phylogeny, the isolate of C. abietis (CBS 125235) and the two strains of an undescribed ascomycete (CBS 125234 and PRM 899855) grouped within the strongly supported monophyletic clade III. The molecular data confirm that C. abietis is not related to other species currently referred to Ceratosphaeria of the Magnaporthaceae. Instead, it is shown on a well supported branch (88/1.0) as a sister of the Papulosaceae. The two strains of the undescribed ascomycete appear on a separate monophyletic branch (100/1.0) without closer relationship to other morphologically similar fungi of the Sordariomycetidae. TAXONOMY Ceratolenta Re´blova´, gen. nov. MycoBank MB800338 Perithecia nonstromatic, immersed, subglobose to globose, dark brown; necks cylindrical emerging above the substratum, straight or slightly curved, ostiolum periphysate. Perithecial wall leathery, two-layered. Paraphyses hyaline, septate, tapering toward the tip, longer than the asci. Asci unitunicate, cylindricalclavate, eight-spored, broadly rounded at the apex containing a refringent inamyloid apical ring, longstipitate. Ascospores fusiform, hyaline, three-septate, obliquely uniseriate or biseriate within the ascus. Etymology: Derived from the generic names Ceratosphaeria and Lentomitella, referring to an intermediate form of both phenotypes. Typus: Ceratolenta caudata Re´blova´ Ceratolenta caudata Re´blova´, sp. nov. FIGS. 2–14 MycoBank MB800339 Perithecia solitary, immersed in the substratum with only necks emerging above substratum, venter subglobose to globose, dark brown, laterally pinched, 370– 450 mm high, 350–420 mm diam. Necks central, cylindrical, upright, straight to slightly flexuous, 750– 1500 mm long, 75–90(–100) mm wide, the wall at the tip of the neck thinner, becoming reddish in transmitted light. Perithecial wall leathery, two-layered, 25–35 mm thick; outer layer of textura prismatica comprising brown polyhedral cells, which are darker toward the surface; inner layer of hyaline, thin-walled, elongated cells. Ostiolum periphysate. Paraphyses hyaline, sep-

tate, constricted at the septa, at maturity partially dissolving, wider near the base ca. 4.5–7.0 mm, tapering to ca. 2.5–3.0 mm toward the tip, longer than the asci. Asci cylindrical-clavate, (90–)110–130 3 9.0–11.0(– 12.0) mm (mean 6 SE 5 114 6 2.3 3 10.7 6 0.1 mm), (70–)82–90(–97) mm long in pars sporifera (mean 6 SE 5 86 6 1.4), stipe (17–)22–39 mm long, tapering, base bulbose, at maturity dissolving, asci floating freely in the centrum, ascal apex obtuse with a donut-like to wedge-shaped, refringent apical annulus 3.5–4.0 mm wide, 2.0–2.3 mm high. Ascospores ellipsoidal, sometimes slightly flattened at one side, tapering at the ends, (13.5–)14.0–15.5(–16.5) 3 5.5–6.5 mm (mean 6 SE 5 14.6 6 0.1 3 6.2 6 0.04 mm), hyaline, smoothwalled, three-septate, obliquely uni- or biseriate, often as a combination of both arrangements, immature and mature ascospores can occur together within an ascus. Colonies in vitro after 30 d on PCA at 25 C in the dark 5–6 mm diam, slow-growing, velvety; aerial mycelium dense, pale brown to gray; margin entire, submerged; reverse brown. Mycelium sterile. Chlamydospores not observed. Etymology: Caudatus (L) with a long tail, referring to a long ascus stipe. Specimens examined: CZECH REPUBLIC. Southern Bohemia: Sˇumava Mountains. National park, Stozˇec, Stozˇecka´ ska´la Mount, on decaying deciduous wood, 15 Sep 2007, M.R. 3008 (HOLOTYPE PRM 899854; living culture CBS ˇ elezna´ Ruda, glacial cirque of the 125234, ex-type). Ibid., Z ˇ erne´ jezero lake National Nature Reserve, on decaying C deciduous wood, 22 Oct 1996, M.R. 895; ibid., on decaying wood of Fagus sylvatica, 13 Aug 1999, M.R. 1802. Ibid., ˇ ertovo jezero lake National Nature glacial cirque of the C Reserve, on decaying wood of Fagus sylvatica, 12 Aug 1999, M.R. 1531, M.R. 1587, M.R. 1597; ibid., 21 Aug 2000, M.R. 2440, M.R. 2510. Ibid., Stozˇec, Spa´lenisˇteˇ Mount, on decaying wood of Sorbus aucuparia, 12 Nov 2000, M.R. 2518; ibid., on decaying wood of Acer pseudoplatanus, 1 Sep 2007, M.R. 2980. Ibid., Stozˇec, Medveˇdice Nature Reserve, on decaying wood of Acer pseudoplatanus, 17 Sep 2007, M.R. 2982 (PRM 899855), M.R. 2984. Novohradske´ hory ˇ ofıńsky´ prales Nature Reserve, on Mountains, Pivonice, Z decaying wood of Fagus sylvatica, 20 May 1997, M.R. 916.

Comments: In some collections, including the specimen PRM 899855, whose DNA sequence data also were analyzed, the length of ascospores occasionally can be slightly longer (up to 17.5 mm long) than it is described based on the type. The ascus stipe is elastic, in squash microscopic preparation it can elongate significantly while still connecting the ascus with the hymenium (FIG. 10). Platytrachelon Re´blova´, gen. nov. MycoBank MB800340 Perithecia nonstromatic, immersed to semi-immersed becoming superficial, subglobose to globose, dark


467

FIGS. 2–14. Ceratolenta caudata. 2–5. Perithecia, venter immersed in decaying wood, long neck emerging. 6. Top of the neck. 7. Longitudinal section of the perithecial wall. 8, 9. Ascospores. 10–14. Asci. Arrow indicates prolonging stipe in an ascus, which is still attached to the hymenium. 2, 7, 11, 12 from PRM 899854; 3, 6, 8 from M.R. 2980; 4, 5, 9 from PRM 899855; 10, 14 from M.R. 2440. Bars: 2–5 5 500 mm; 6 5 100 mm; 7 5 25 mm; 8–14 5 10 mm.

brown to black, with sparse brown hyphae; necks cylindrical, laterally pinched on drying, straight or slightly curved, ostiolum periphysate. Perithecial wall leathery, two-layered. Paraphyses hyaline, septate, tapering toward the tip, longer than the asci. Asci unitunicate, cylindrical-clavate, eight-spored, broadly rounded at the apex, with a refringent inamyloid annulus, longstipitate. Ascospores fusiform, hyaline, 3–5-septate, uniseriate within the ascus.

Etymology: Platy´s (Gk) flattened, trachelos (Gk) neck; referring to the flattened neck of perithecia. Typus: Platytrachelon abietis (Re´blova´) Re´blova´ Platytrachelon abietis (Re´blova´) Re´blova´, comb. nov. FIGS. 15–40 Basionym: Ceratosphaeria abietis Re´blova´, Sydowia 50:236. 1998. MycoBank MB800341

468

MYCOLOGIA

FIGS. 15–26. Platytrachelon abietis. 15–17. Perithecia on the natural substratum. 18, 19. Asci with most of the ascospores normally developed. 20. Ascus with only four mature and abnormally large ascospores. Arrows indicate immature ascospores within the asci. 21. Longitudinal section of the perithecial wall. 22. Abnormally developed ascospores, swollen at the septa. 23. Ascospores of regular size. 24. Paraphyses. 25. Apical annulus. 26. Bottom part of the ascus stipe. Arrows indicate where the outer wall disintegrates. 15, 17, 20, 22 from PRM 842974; 16, 18, 19, 21, 23–26 from M.R. 2970. Bars: 15–17 5 1000 mm; 21 5 50 mm; 20, 26 5 20 mm; 18, 19, 22–25 5 10 mm.

Perithecia solitary to gregarious, superficial, semiimmersed to immersed in the substratum with only necks emerging, venter subglobose to globose, dark brown to black, laterally pinched, 430–580 mm high, 420–600 mm diam. Hyphal subiculum consisting of sterile, branched, grayish-brown hyphae 3.5–4.5 mm wide cover the venter, bottom part of the neck and

surface of the substrate around the perithecia when they are superficial or semi-immersed. Necks prominent, central, cylindrical, upright, straight to slightly curved, becoming laterally pinched upon drying, obtuse at the tip with a distinct ostiolum, up to 1500 mm long, 200–240 mm wide. Perithecial wall leathery, two-layered, 55–75 mm thick; outer layer of


469

FIGS. 27–40. Anamorph and synanamorph of Platytrachelon abietis CBS 125235 (PCA, 30 d). 27–33. Conidia and subglobose conidiogenous cells. 34–39. Conidiogenous cells of the synanamorph. 40. Conidia of the synanamorph. Bars: 27–33 5 20 mm, 34– 40 5 10 mm.

textura prismatica comprising brown polyhedral cells, which are darker toward the surface; inner layer of hyaline, thin-walled, elongated and flattened cells. Ostiolum periphysate. Paraphyses hyaline, cylindrical, branched, septate, slightly constricted at the septa, wider near the base, ca. (3.0–)4.0–5.5 mm, tapering to ca. 2.0–2.5 mm toward the tip, longer than the asci. Asci cylindrical-clavate, 158–200 mm long in pars sporifera (mean 6 SE 5 177 6 3.0), 8.0–9.5 mm wide (mean 6 SE 5 8.7 6 0.1 mm), stipe 84–120 mm long, slender, tapering, base bulbose, at maturity dissolving along the sides, ascal apex obtuse with a refringent wedge-shaped apical annulus 3.5–4.0 mm wide, 1.5– 2.2 mm high. Ascospores fusiform, sometimes slightly flattened at one side, (19–)22–27(–28) 3 5.5–6.0 mm (mean 6 SE 5 23.5 6 0.4 3 5.6 6 0.05 mm), hyaline, smooth-walled, 3(–5)-septate, constricted at the septa,

uniseriate. Abnormally developed ascospores 31–41(– 45) 3 (6.5–)7.5–8.0 mm, sometimes with end and middle cells inflated. Within an ascus often four immature and four mature and larger ascospores occur together. Colonies in vitro after 30 d on PCA at 25 C in the dark 9–10 mm diam, felt-like; aerial mycelium dense, brown with a pale brown-gray marginal zone; margin entire; reverse dark brown. Sporulating conidiophores developing throughout the colony in 30 d on PCA at 25 C in the dark. Conidiogenous cells 3.5– 4.5 3 3–3.5 mm (mean 6 SE 5 3.8 6 0.1 3 3.3 6 0.04 mm), spherical, hyaline to subhyaline, blastic, arising vertically from subhyaline hyphae, proliferating sympodially, often one to several cells in a row. Conidia 11.5–14(–16) 3 4.0–5.0 mm (mean 6 SE 5 13.0 6 0.4 3 4.7 6 0.1 mm), ellipsoidal, pale brown,

470

MYCOLOGIA

smooth-walled, one-septate, not constricted at the septum, occasionally arising directly from subhyaline or pigmented hyphae. Conidiogenous cells of the synanamorph 6.0–7.5 3 2.5–3.0 mm (mean 6 SE 5 6.9 6 0.5 3 2.9 6 0.1 mm), cylindrical to ampulliform, one-celled, hyaline to subhyaline, terminating in a single phialidic opening or with additional lateral opening. Collarette 0.5–1.0 mm high, ca. 2.5 mm wide, shallow wedge-shaped, hyaline, indistinct. Conidia 5.5–6.5(–8.0) 3 2.5–3.0 mm (mean 6 SE 5 6.6 6 0.4 3 0.8 6 0.01 mm), cylindrical, strongly curved, hyaline, one-celled, smooth-walled. Chlamydospores not observed. Specimens examined: CZECH REPUBLIC. Southern ˇ elezna´ Ruda, Bohemia: Sˇumava Mountains National park, Z ˇ erne´ jezero lake, on the inner side of glacial cirque of the C decaying bark of Abies alba, 22 Oct 1996, M.R. (HOLOTYPE PRM 842974); ibid. M.R. 870, M.R. 888; ibid., Za´ton ˇ , Jilmova´ ska´la Mount Nature Monument, on the inner side of the decaying bark of Abies alba, 3 Sep 2007, M.R. 2970 (living strain CBS 125235). UKRAINE. Ruthenia: Carpathian Mountains, Borzˇava Massif, Guklivij, on the inner side of decaying bark of Abies alba, 21 Jul 1998, M.R. 1243.

Comments: At the time this species was introduced as Ceratosphaeria abietis, the description based on the type (PRM 842974) was in agreement with observations made on two additional collections (Re´blova´ 1998). All three specimens originated from one ˇ erne´ jezero lake is locality. The glacial cirque of the C a National Nature Reserve, a unique place with near natural forest ecosystems and occurrence of rare plants and animals at 1006 m. In the dominant spruce-fir forest, the research plot was located in a small area of beech wood with mixture of Abies alba, Acer pseudoplatanus, Sorbus aucuparia and Lonicera nigra. In all three collections the ascospores showed abnormal growth; within the ascus four ascospores were predominantly developed and over-sized at the expense of four others, which remained immature and small (FIGS. 20, 22). In material from different localities from the Czech Republic and Ukraine (Carpathian Mountains), although from similar mountain beech forests mixed with Abies alba, this deviation was not observed or was less prominent (FIGS. 18, 19). Therefore, the fungus is redescribed based on recently collected material and observations in vitro. Based on the so far collected material, the fungus is known only from decaying wood and inner side of decaying bark of Abies alba. The length of the whole ascus, 260–310 mm (mean 6 SE 5 276 6 6.4 mm), can vary significantly. It depends on the length of the stipe, whose outer layer disintegrates at maturity along the sides and the inner layer possesses the ability to stretch, often up to

100 mm. This phenomenon cannot usually be seen in asci, which are floating freely in the centrum and in which the lower part of the stipe is already broken. Only stipes of mature asci, which are still attached with the bulbose base to the hymenium, can show this effect. KEY TO FUNGI CHARACTERIZED BY DARK BEAKED ASCOMATA AND HYALINE TO SUBHYALINE ASCOSPORES INCLUDED IN THE CLADE III OF THE

SORDARIOMYCETIDAE

Families Annulatascaceae s. str. and Magnaporthaceae, which accommodate fungi morphologically similar to Ceratolenta, Platytrachelon and whose members grouped in Clade III, are not included in the key. Images of observed and studied taxa with morphologically similar characteristics are included for comparison (FIGS. 41–59). For a discussion and distinction of Annulusmagnus J. Campb. & Shearer (FIGS. 41–50), Annulatascus K.D. Hyde, but also Pseudoproboscispora Punith. (FIGS. 42, 47) and other freshwater taxa currently referred to the Annulatascaceae s. str. see e.g. Hyde (1992), Wong et al. (1998), Campbell and Shearer (2004), Campbell et al. (2003), Vijaykrishna et al. (2006), Abdel-Wahab et al. (2011), and Boonyuen et al. (2012). For a discussion and distinction of Ceratosphaeria (FIGS. 43, 44, 48), Ceratosphaerella Huhndorf et al., Muraeriata Huhndorf et al., Ophioceras Sacc., Pseudohalonectria Minoura & T. Muroi (FIGS. 45, 46, 49) and other morphologically similar fungi currently referred to the Magnaporthaceae (see Hyde et al. [1997], Huhndorf et al. [2008], Thongkantha et al. [2011], Zhang et al. [2011]). The saprobic lignicolous fungi of the latter family are characterized by dark ascomata with carbonized walls, long upright necks, scolecosporous, cylindrical or fusiform often naviculate ascospores, which are overlapping in a fascicle or are 3–4-seriate within the short-stipitate ascus; the apical annulus is inamyloid, tall wedge-shaped. 1.

1.

3. 3.

Ascospores scolecosporous or cylindrical, overlapping in a fascicle within the ascus; ascomata deeply immersed under a black clypeus . . . Neolinocarpon Ascospores and ascomata not as above . . . . . . . . . 2 2. Ascomata with papilla or beak or short conical neck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 2. Ascomata with globose or subglobose venter, neck cylindrical or filiform . . . . . . . . . . . . . 6 Ascomata subglobose or globose, papillate; ascospores ellipsoidal . . . . . . . . Cryptadelphia (FIG. 51) Ascomata pyriform or subglobose to conical with a short conical neck . . . . . . . . . . . . . . . . . . . . . . . . 4 4. Ascospores with bifurcate polar appendages . . . . . . . . . . . . . . . . . . . . . . . . Fluminicola 4. Ascospores without appendages or sheath . . 5

RE´BLOVA´: TWO TAXONOMIC NOVELTIES IN SORDARIOMYCETIDAE 5.

5.

7. 7.

9.

9.

11. 11.

13. 13.

Ascospores asymmetrically fusiform, clavate or broadly ellipsoid, muriform, obliquely uniseriate; asci short-stipitate . . . . . . . . . Rhamphoria (FIG. 52) Ascospores fusiform, three-septate, uniseriate; asci with a long disintegrating stipe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rhodoveronaea (FIG. 53) 6. Ascomata with filiform necks; interthecial filaments lacking; asci formed in chains, evanescent; ascospores aseptate, with eccentric wall thickening or sheaths . . . . . . . Ophiostoma 6. Ascomata, asci and ascospores not as above . 7 Ascospores distoseptate . . . . . . . . . . . . . Vertexicola Ascospores with true septa . . . . . . . . . . . . . . . . . 8 8. Asci short-stipitate, stipe rounded to obtuse . 9 8. Asci with a long tapering stipe . . . . . . . . . . 10 Ascospores fusiform; apical annulus staining blue with aqueous cotton blue and aqueous nigrosin . . . . . . . . . . . . . . . . . . . . . . . . . Cyanoannulus Ascospores ellipsoidal; apical annulus not staining blue . . . . . . . . . . . . . . . . . . . Lentomitella (FIG. 55) 10. Ascospores ellipsoidal, oblong, suballantoid to horseshoe-shaped; apical annulus minute . 11 10. Ascospores fusiform; apical annulus wedgeshaped . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Ascospores nonseptate; asci in pars sporifera longer than 35 mm . . . . . . . . . . . . . . . . . . . . . . 12 Ascospores nonseptate or septate; asci in pars sporifera up to 20–35 mm long . . . . . . . . . . . . . 13 12. Ascospores oblong, 2–3-seriate; terrestrial habitat . . . . . . . . . . . . . . . . Natantiella (FIG. 54) 12. Ascospores ellipsoidal, uniseriate or obliquely uniseriate to biseriate; freshwater habitat . . . . . . . . . . . . . . . . . . . . . . . . . . . . Aquaticola Ascospores oblong to suballantoid. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Barbatosphaeria (FIGS. 56, 58) Ascospores horseshoe-shaped. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tectonidula (FIGS. 57, 59) 14. Ascospores three-septate, ellipsoidal-fusiform, obliquely uni- or biseriate, often arranged in the middle of the sporiferous part . . . . . . . . . . . . . . Ceratolenta (FIGS. 2–14) 14. Ascospores 3–5-septate, fusiform, uniseriate, sometimes only four mature ascospores developing . . . . . . . . . . Platytrachelon (FIGS. 15–40)

DISCUSSION In the phylogram (FIG. 1), Ceratolenta and Platytrachelon represent two novel lineages in the Sordariomycetidae. The simple morphological characters of their ascomata with long necks, carbonaceous walls, stipitate asci and hyaline, septate ascospores may cause difficulties in classifying them without molecular data and in vitro studies. The main reason why these genera remained unrecognized was that their morphology is unexceptional compared with other morphologically similar taxa. The robust clade III, where both genera

471

grouped, is divided into two subclades. The Magnaporthaceae, whose members are linked with phialidic or tretic conidiogenesis (Bussaban et al. 2005, Huhndorf et al. 2008, Thongkantha et al. 2009, Zhang et al. 2011), and another large, more heterogeneous group. The latter clade contains the Ophiostomatales, the three families Annulatascaceae s. str., Papulosaceae and Thyridiaceae, both new genera and several other monotypic or small genera with uncertain phylogenetic relationships. The majority of these fungi share similar hyaline, ellipsoidal to fusiform, oblong ascospores in cylindrical-clavate stipitate asci, dark ascomata with carbonized walls and conical, cylindrical to filiform necks or beaks. The conidiogenesis (if known) is holoblastic, often denticulate. Ceratolenta and Platytrachelon are morphologically similar to Fluminicola, Rhodoveronaea and some members of the otherwise still polyphyletic Annulatascaceae (based on DNA data, e.g. Campbell et al. 2003, Abdel-Wahab et al. 2011, Boonyuen et al. 2012). Ceratolenta most closely resembles Rhodoveronaea. They share ellipsoidal to fusiform, three-septate, hyaline ascospores in cylindrical asci with a long tapering and partially dissolving stipe. The main difference between them lies in the arrangement of the ascospores within the ascus and in the perithecia; in Rhodoveronaea the ascospores are uniseriate and the neck of perithecia is conical and distinctly tapering. While the living culture of Ceratolenta produces only sterile mycelium, the anamorph of Rhodoveronaea produces holoblastic conidia on integrated polyblastic conidiogenous cells on dematiaceous conidiophores. Molecular data suggest they are not closely related. The recently described Conlarium dumplumascospora F. Liu & L. Cai (Liu et al. 2012) of the Sordariomycetidae is similar to Ceratolenta in having dark, nonstromatic globose to subglobose ascomata with long necks, carbonaceous walls and septate hyaline ascospores. It differs from the C. caudata by short-stipitate asci with a relatively large, tall wedge-shaped apical annulus and ascospores with polar appendages. The simple phenotype of Ceratosphaeria abietis could be deceiving. When the species was described, the genus was still a heterogeneous collection of various species. A detailed morphological characterization of Ceratosphaeria lampadophora, the type species of the genus, was published by Hyde et al. (1997). Based on nc28S sequence data, C. lampadophora was shown to belong to the Magnaporthaceae (Re´blova´ et al. 2004). Later, another species, Ceratosphaeria phialidica (Shearer) Huhndorf et al., and two of the Ceratosphaeria segregates, Ceratosphaerella and Muraeriata, also were accommodated in the Magnaporthaceae (Huhndorf et al. 2008). They are characterized

472

MYCOLOGIA

FIGS. 41–52. Six similar genera. 41, 50. Annulusmagnus (A. triseptatus). 41. Perithecia. 50. Ascus with ascospores. 42, 47. Pseudoproboscispora (P. caudae-suis). 42. Perithecia. 47. Long-stipitate asci with ascospores. 43, 44, 48. Ceratosphaeria (C. lampadophora). 43. Perithecia. 44. Longitudinal section of perithecia. 48. Asci with ascospores and distinct tall wedge-shaped apical annulus. 45, 46, 49. Pseudohalonectria (P. lutea, FIG. 46 Pseudohalonectria sp.). 45, 46. Longitudinal section of perithecia. 49. Ascus. 51. Cryptadelphia (C. brevior), asci with ascospores. 52. Rhamphoria (R. pyriformis), ascus containing eight muriform


473

FIGS. 53–59. Five similar genera. 53. Rhodoveronaea (R. varioseptata), long-stipitate ascus with ascospores. 54. Natantiella (N. ligneola), ascus. 55. Lentomitella (L. crinigera), ascus with ascospores. 56, 58. Barbatosphaeria. (B. barbirostris). 56. Asci filled with oblong ascospores. 58. Perithecia growing on decaying wood under the periderm. 57, 59. Tectonidula (T. hippocrepida). 57. Asci filled with horseshoe-shaped ascospores. 59. Perithecia growing on decaying wood under the periderm. 53 from M.R. 1818; 54 from M.R. 3612; 55 from M.R. 1671; 56 from J.F. 04053, 58 from M.R. 698; 57, 59 from PDD 81440. Bars: 58, 59 5 250 mm; 53–56 5 10 mm; 57 5 5 mm.

by cylindrical to filiform or narrowly fusiform often naviculate, hyaline, septate or nonseptate ascospores arranged 2–4-seriately within short-stipitate asci. The conidiogenesis is phialidic in Ceratosphaeria (anamorph harpophora-like) or monotretic in Ceratosphaerella (anamorph didymobotryum-like) (Re´ blova´ 2006, Huhndorf et al. 2008). However 38 additional species including synonyms are currently referred to Ceratosphaeria based on the simple teleomorph morphology. Their systematic placement needs to be verified in recollected material with molecular data and cultivation studies. Platytrachelon abietis grouped as a sister of the Papulosaceae clade. That clade includes four other

fungi, of which three are known exclusively from freshwater or marine habitats. Among them Fluminicola bipolaris S.W. Wong et al. is reminiscent of Platytrachelon in three-septate, fusiform ascospores arranged uniseriately or obliquely uniseriately in cylindrical asci with a long tapering stipe and wedgeshaped apical annulus. It differs from Platytrachelon in the ascospores with bifurcate polar appendages. The dematiaceous anamorph of Platytrachelon was formed only in culture. It resembles the triadelphialike anamorph linked to the freshwater ascomycete Ascolacicola aquatica Ranghoo & K.D. Hyde (as Trichocladium uniseptatum [Berk. & Broome] S. Hughes & Piroz., Ranghoo and Hyde 1998), for

r ascospores, which during maturation produce ascoconidia filling the entire ascus. 41, 50 from J.F. 06336; 42, 47 from J.F. 06234; 43, 44, 48 from J.F. 08009; 45, 49 from J.F. 06157; 46 from J.F. 06252; 51 from PRM 647191; 52 from M.R. 3642. Bars: 41–46 5 250 mm; 50 5 50 mm; 47–49 5 20 mm; 51, 52 5 10 mm.

474

MYCOLOGIA

which Ranghoo et al. (1999) assumed a relationship with the Sordariales based on a nc28S sequence. However, a re-examination of the same nc28S sequence (AF132322) by the author shows that this is probably not a sequence of the correct fungus, instead suggesting a relationship to the Chaetothyriales of the Eurotiomycetes (not shown). To clarify the phylogenetic relationship between Platytrachelon and Triadelphia uniseptata (Berk. & Broome) P.M. Kirk, we analyzed another nc28S sequence of the latter species from Canada. Their close relationship was not confirmed because molecular data placed T. uniseptata in the Hypocreomycetidae, Sordariomycetes (Re´blova´ and Seifert unpubl). ACKNOWLEDGMENTS This study was supported by the Project of the National Foundation of the Czech Republic (GAP 506/12/0038) and as a long-term research development project of the Institute of Botany, Academy of Sciences No. RVO 67985939. I thank Vaćlav Sˇteˇpańek, who undertook the DNA sequencing of isolates of Ceratolenta and Platytrachelon and other newly sequenced fungi. I am grateful for the help of Jacques Fournier for his photographs of selected lignicolous ascomycetes, their ascomata, longitudinal sections and asci (FIGS. 41–47). I wish to thank Walter Gams for carefully reading the manuscript and further suggestions.

LITERATURE CITED Abdel-Wahab MA, Abdel-Aziz FA, Mohamed SS, Abdel-Aziz AE. 2011. Annulatascus nilensis sp. nov., a new freshwater ascomycete from the river Nile, Egypt. IMA Fungus 2:1–6, doi:10.5598/imafungus.2011.02.01.01 Boonyuen N, Sri-indrasutdhi V, Suetrong S, Jones EBG. 2012. Annulatascus aquatorba sp. nov., a lignicolous freshwater ascomycete from Sirindhorn Peat Swamp Forest, Narathiwat, Thailand. Mycologia 104:746–757, doi:10.3852/11-238 Bussaban B, Lumyong S, Seelanan T, Park DC, McKenzie EHC, Hyde KD. 2005. Molecular and morphological characterization of Pyricularia and allied genera. Mycologia 97:1002–1011, doi:10.3852/mycologia.97.5.1002 Campbell J, Shearer CA. 2004. Annulusmagnus and Ascitendus, two new genera in the Annulatascaceae. Mycologia 96:822–833, doi:10.2307/3762115 ———, ———, Crane JL, Fallah PM. 2003. A reassessment of two freshwater ascomycetes, Ceriospora caudae-suis and Submersisphaeria aquatica. Mycologia 95:41–53, doi:10.2307/3761960 Cunningham CW. 1997. Can three incongruence tests predict when data should be combined? Mol Phylogenet Evol 14:733–740. Ferrer A, Miller AN, Sarmiento C, Shearer C. 2012. Three new genera representing novel lineages of Sordariomycetidae (Sordariomycetes, Ascomycota) from tropical

freshwater habitats in Costa Rica. Mycologia 104:865– 879, doi:10.3852/11-111 Gams W, Hoekstra ES, Aptroot A. 1998. CBS course of mycology. 4th ed. Baarn, The Netherlands: Centraalbureau voor Schimmelcultures. 165 p. Gutell RR. 1993. Collection of small subunit (16S- and 16Slike) ribosomal RNA structures. Nucleic Acids Res 21: 3051–3054, doi:10.1093/nar/21.13.3051 ———, Gray MW, Schnare MN. 1993. A compilation of large subunit (23S and 23S-like) ribosomal RNA structures. Nucleic Acids Res 21: 3055– 3074, doi:10.1093/nar/21.13.3055 Hall TA. 1999. BioEdit 5.0.9: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98. Ho WWH, Tsui CKM, Hodgkiss IJ, Hyde KD. 1999. Aquaticola, a new genus of Annulatascaceae from freshwater habitats. Fungal Divers 3:87–97. Huelsenbeck JP, Ronquist F. 2001. MrBayes: Bayesian inference of phylogenetic trees. Bioinformatics 17: 754–755, doi:10.1093/bioinformatics/17.8.754 Huhndorf SM, Greif M, Mugambi GK, Miller AN. 2008. Two new genera in the Magnaporthaceae, a new addition to Ceratosphaeria and two new species of Lentomitella. Mycologia 100:940–955, doi:10.3852/08-037 Hyde KD. 1992. Tropical Australian freshwater fungi I. Annulatascus velatispora gen. et sp. nov., A. bipolaris sp. nov. and Nais aquaticola sp. nov. (Ascomycetes). Austr Syst Bot 5:117–124, doi:10.1071/SB9920117 ———, Read SJ, Jones EBG, Moss ST. 1997. Tropical Australian freshwater fungi XII. Rivulicola incrustata gen. et sp. nov. and notes on Ceratosphaeria lampadophora. Nova Hedwigia 64:185–196. Larget B, Simon DL. 1999. Markov chain Monte Carlo algorithms for the Bayesian analysis of phylogenetic trees. Mol Biol Evol 16:750–759, doi:10.1093/oxfordjournals.molbev.a026160 Liu F, Hu DM, Cai L. 2012. Conlarium duplumascospora gen. et. sp. nov. and Jobellisia gregariusca sp. nov. from freshwater habitats in China. Mycologia (In press), doi:10.3852/11-379. Nylander J. 2008. MrModeltest2 2.3 (program for selecting DNA substitution models using PAUP*). Uppsala, Sweden: Evolutionary Biology Centre. Raja HA, Campbell J, Shearer CA. 2003. Freshwater ascomycetes: Cyanoannulus petersenii, a new genus and species from submerged wood. Mycotaxon 88:1– 17. Ranghoo VM, Hyde KD. 1998. Ascolacicola aquatica gen. et sp. nov. and a new species of Ascotaiwania from wood submerged in a reservoir in Hong Kong. Mycologia 90: 1055–1062, doi:10.2307/3761280 ———, ———, Liew ECY, Spatafora JW. 1999. Family placement of Ascotaiwania and Ascolacicola based on DNA sequences of the large subunit rRNA gene. Fungal Divers 2:159–168. ———, ———, Wong SW, Tsui CKM, Jones EBG. 2000. Vertexicola caudatus gen. et sp. nov. and a new species of Rivulicola from submerged wood in freshwater

RE´BLOVA´: TWO TAXONOMIC NOVELTIES IN SORDARIOMYCETIDAE habitats. Mycologia 92:1019–1026, doi:10.2307/ 3761596 Re´blova´ M. 1998. Fungal diversity in the Czech Republic. New species of Apiorhynchostoma, Capronia, Ceratosphaeria and Lasiosphaeria. Sydowia 50:229–251. ———. 2006. Molecular systematics of Ceratostomella sensu lato and morphologically similar fungi. Mycologia 98: 68–93, doi:10.3852/mycologia.98.1.68 ———. 2009. Teleomorph of Rhodoveronaea (Sordariomycetidae) discovered and re-evaluation of Pleurophragmium. Fungal Divers 36:129–139. ———, Gams W, Seifert KA. 2011. Monilochaetes and allied genera of the Glomerellales, and a reconsideration of families in the Microascales. Stud Mycol 68:163–191, doi:10.3114/sim.2011.68.07 ———, Mostert L, Gams W, Crous PW. 2004. New genera in the Calosphaeriales: Togniniella and its anamorph Phaeocrella, and Calosphaeriophora as anamorph of Calosphaeria. Stud Mycol 50:533–550. ———, Sˇteˇpańek V. 2009. New fungal genera, Tectonidula gen. nov. for Calosphaeria-like fungi with holoblasticdenticulate conidiogenesis and Natantiella gen. nov. for three species segregated from Ceratostomella. Mycol Res 113:991–1002, doi:10.1016/j.mycres.2009. 06.003 Saccardo PA. 1878. Fungi Veneti novi vel critici IX. Michelia 1:361–445. Shearer CL, Crane JL, Fallah PM. 2003. A reassessment of two freshwater ascomycetes, Ceriospora caudae-suis and Submersisphaeria aquatica. Mycologia 95: 41– 53, doi:10.2307/3761960 Stamatakis A. 2006. RAxML-VI-HPC: maximum likelihoodbased phylogenetic analyses with thousands of taxa and

475

mixed models. Bioinformatics 22: 2688– 2690, doi:10.1093/bioinformatics/btl446 Swofford DL. 2002. PAUP* 4.0b10: phylogenetic analysis using parsimony (*and other methods). Sunderland, Massachusetts: Sinauer Associates. Thongkantha S, Jeewon R, Vijaykrishna D, Lumyong S, McKenzie EHC, Hyde KD. 2009. Molecular phylogeny of Magnaporthaceae (Sordariomycetes) with a new species Ophioceras chiangdaoense from Dracaena loureiroi in Thailand. Fungal Divers 34:157–173. Tsui CKM, Hodgkiss IJ, Hyde KD. 2003. Three new species of Aquaticola (Ascomycetes) from tropical freshwater habitats. Nova Hedwigia 77:161–168, doi:10.1127/00295035/2003/0077-0161 Vijaykrishna D, Jeewon R, Hyde KD. 2006. Molecular taxonomy, origins and evolution of freshwater ascomycetes. Fungal Divers 23:351–390. von Ho¨hnel F. 1905. Mykologische Fragmente. Ann Mykol 3:548–560. von Niessl G. 1876. Notizen u¨ber neue und kritische Pyrenomyceten. Verh Naturf Ver Bru¨nn 14:165–218. Wong SW, Hyde KD. 1999. Proboscispora aquatica gen. et sp. nov., from wood submerged in freshwater. Mycol Res 103:81–87, doi:10.1017/S0953756298006820 ———, ———, Fro¨hlich J. 1998. Annulatascaceae, a new ascomycete family from the tropics. Syst Ascom 16:17–25. ———, ———, Jones EBG. 1999. Ultrastructural studies of freshwater ascomycetes, Fluminicola bipolaris gen. et sp. nov. Fungal Divers 2:189–197. Zhang N, Zhao S, Shen Q. 2011. A six-gene phylogeny reveals the evolution of mode of infection in the rice blast fungus and allied species. Mycologia 103:1267– 1276, doi:10.3852/11-022