HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Free via Open Access: OA OA Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Crous, P.W. Articles by Groenewald, J.Z. Search for Related Content PubMed PubMed Citation Articles by Crous, P.W. Articles by Groenewald, J.Z.

Opportunistic, human-pathogenic species in the Herpotrichiellaceae are phenotypically similar to saprobic or phytopathogenic species in the Venturiaceae

¹ CBS Fungal Biodiversity Centre, P.O. Box 85167, 3508 AD, Utrecht, The Netherlands
² Botanische Staatssammlung München, Menzinger Strasse 67, D-80638 München, Germany
³ Martin-Luther-Universität, Institut für Biologie, Geobotanik und Botanischer Garten, Herbarium, Neuwerk 21, D-06099 Halle, Germany
⁴ CSIRO Food Science Australia, 11 Julius Avenue, North Ryde, NSW 2113, Australia
⁵ Division of Environmental Science & Ecological Engineering, Korea University, Seoul 136-701, Korea

^* Correspondence: Pedro W. Crous, p.crous{at}cbs.knaw.nl

	Abstract

TOP Abstract INTRODUCTION MATERIALS AND METHODS RESULTS Members of Venturiaceae Excluded taxa DISCUSSION References

 
Although morphologically similar, species of Cladophialophora (Herpotrichiellaceae) were shown to be phylogenetically distinct from Pseudocladosporium (Venturiaceae), which was revealed to be synonymous with the older genus, Fusicladium. Other than being associated with human disorders, species of Cladophialophora were found to also be phytopathogenic, or to occur as saprobes on organic material, or in water, fruit juices, or sports drinks, along with species of Exophiala. Caproventuria and Metacoleroa were confirmed to be synonyms of Venturia, which has Fusicladium (= Pseudocladosporium) anamorphs. Apiosporina, based on A. collinsii, clustered basal to the Venturia clade, and appears to represent a further synonym. Several species with a pseudocladosporium-like morphology in vitro represent a sister clade to the Venturia clade, and are unrelated to Polyscytalum. These taxa are newly described in Fusicladium, which is morphologically close to Anungitea, a heterogeneous genus with unknown phylogenetic affinity. In contrast to the Herpotrichiellaceae, which were shown to produce numerous synanamorphs in culture, species of the Venturiaceae were morphologically and phylogenetically more uniform. Several new species and new combinations were introduced in Cladophialophora, Cyphellophora (Herpotrichiellaceae), Exophiala, Fusicladium, Venturia (Venturiaceae), and Cylindrosympodium (incertae sedis).

Taxonomic novelties: Cladophialophora australiensis Crous & A.D. Hocking, sp. nov., Cladophialophora chaetospira (Grove) Crous & Arzanlou, comb. nov., Cladophialophora hostae Crous, U. Braun & H.D. Shin, sp. nov., Cladophialophora humicola Crous & U. Braun, sp. nov., Cladophialophora potulentorum Crous & A.D. Hocking, sp. nov., Cladophialophora scillae (Deighton) Crous, U. Braun & K. Schub., comb. nov., Cladophialophora sylvestris Crous & de Hoog, sp. nov., Cylindrosympodium lauri Crous & R.F. Castañeda, sp. nov., Cyphellophora hylomeconis Crous, de Hoog & H.D. Shin, sp. nov., Exophiala eucalyptorum Crous, sp. nov., Fusicladium africanum Crous, sp. nov., Fusicladium amoenum (R.F. Castañeda & Dugan) Crous, K. Schub. & U. Braun, comb. nov., Fusicladium brevicatenatum (U. Braun & Feiler) Crous, U. Braun & K. Schub., comb. nov., Fusicladium fagi Crous & de Hoog, sp. nov., Fusicladium intermedium (Crous & W.B. Kendr.) Crous, comb. nov., Fusicladium matsushimae (U. Braun & C.F. Hill) Crous, U. Braun & K. Schub., comb. nov., Fusicladium pini Crous & de Hoog, sp. nov., Fusicladium ramoconidii Crous & de Hoog, sp. nov., Fusicladium rhodense Crous & M.J. Wingf., sp. nov., Venturia hystrioides (Dugan, R.G. Roberts & Hanlin) Crous & U. Braun, comb. nov.

Keywords Anungitea / Anungitopsis / Cladophialophora / Exophiala / Fusicladium / phylogeny / Pseudocladosporium / systematics / Venturia

	INTRODUCTION

TOP Abstract INTRODUCTION MATERIALS AND METHODS RESULTS Members of Venturiaceae Excluded taxa DISCUSSION References

 
Species of Cladophialophora Borelli are relatively simple hyphomycetes with brown hyphae that give rise to branched chains of pale brown conidia. Phylogenetically they are defined to belong to the Chaetothyriales (Haase et al. 1999, Untereiner 2000), an order containing numerous opportunists (de Hoog et al. 2000); teleomorph relationships are with Capronia Sacc. in the Herpotrichiellaceae. In several cases cladophialophora-like synanamorphs are found accompanying black yeasts of the genus Exophiala J.W. Carmich. (de Hoog et al. 1995). Braun & Feiler (1995) placed several saprobic hyphomycetes in Cladophialophora, and described Capronia hanliniana U. Braun & Feiler as teleomorph of Cladophialophora brevicatenata U. Braun & Feiler. This work was continued by Dugan et al. (1995), who described an additional teleomorph, Capronia hystrioides Dugan, R.G. Roberts & Hanlin for Cladophialophora hachijoensis (Matsush.) U. Braun & Feiler. Untereiner (1997) reduced Capronia hystrioides to synonymy with C. hanliniana, and placed them in Venturia Sacc. (Venturiaceae, Pleosporales). The concept of Cladophialophora hachijoensis, which is based on Phaeoramularia hachijoensis Matsush. (Matsushima 1975) is confused, however, and phylogenetic studies have revealed that isolates attributed to this name in recent studies, were in fact representatives of three different species in phylogenetically distinct genera (Braun et al. 2003). The separation of Cladophialophora with Capronia teleomorphs (Herpotrichiellaceae, Chaetothyriales; commonly isolated as human pathogens), from predominantly saprobic or phytopathogenic isolates in the Dothideomycetes was recognised by Braun (1998). Recently the cactus endophyte Cladophialophora yegresii de Hoog was reported to be the nearest neighbour of C. carrionii (Trejos) de Hoog et al., a major agent of human chromoblastomycosis (de Hoog et al. 2007), so that the main distinction between the two anamorph genera remains in their phylogenetic positions. Capronia hanliniana and C. hystrioides were again recognised as distinct species, and placed in a new genus, Caproventuria U. Braun (Venturiaceae), while their anamorphs were accommodated in Pseudocladosporium U. Braun. Caproventuria was primarily distinguished from Venturia based on its distinct Pseudocladosporium anamorphs. Recently, Crous et al. (2007b) introduced a third genus, namely Sympoventuria Crous & Seifert, which produces a sympodiella-like anamorph in culture. To complicate matters further, Beck et al. (2005) concluded, based on an ITS DNA phylogeny, that the morphology attributed to the form genera Spilocaea Fr., Pollaccia E. Bald. & Cif., and Fusicladium Bonord. has evolved several times within Venturia, and that a single anamorph genus should be used for Venturia, namely Fusicladium (see Schubert et al. 2003 for additional generic synonyms).

In their treatment of Venturia anamorphs, Schubert et al. (2003) excluded Pseudocladosporium, and stated that its status needs to be confirmed along with that of other genera such as Anungitea B. Sutton, Fusicladium and Polyscytalum Riess. In the study by Beck et al. (2005) an isolate of Caproventuria hystrioides (Pseudocladosporium sp.) was included to confirm the link to the Venturiaceae, though this was not well resolved, nor was the status of the older generic names mentioned above addressed. The aim of the present study, therefore, was to use DNA sequence comparisons in conjunction with morphology in an attempt to clarify these generic issues, as well as to determine which morphological characters could be used to distinguish Pseudocladosporium from Cladophialophora.

	MATERIALS AND METHODS

TOP Abstract INTRODUCTION MATERIALS AND METHODS RESULTS Members of Venturiaceae Excluded taxa DISCUSSION References

 
Isolates
Cultures were obtained from the Centraalbureau voor Schimmelcultures (CBS) in Utrecht, the Netherlands, or isolated from plant material incubated in moist chambers to promote sporulation. Isolates were cultured on 2 % malt extract plates (MEA; Gams et al. 2007), by obtaining single conidial colonies as explained in Crous (2002). Colonies were subcultured onto fresh MEA, oatmeal agar (OA), potato-dextrose agar (PDA) and synthetic nutrient-poor agar (SNA) (Gams et al. 2007), and incubated at 25 °C under continuous near-ultraviolet light to promote sporulation.

DNA extraction, amplification and phylogeny
Fungal colonies were established on agar plates, and genomic DNA was isolated following the CTAB-based protocol described in Gams et al. (2007). The primers V9G (de Hoog & Gerrits van den Ende 1998) and LR5 (Vilgalys & Hester 1990) were used to amplify part (ITS) of the nuclear rDNA operon spanning the 3' end of the 18S rRNA gene (SSU), the first internal transcribed spacer (ITS1), the 5.8S rRNA gene, the second ITS region and the 5' end of the 28S rRNA gene (LSU). Four internal primers, namely ITS4 (White et al. 1990), LR0R (Rehner & Samuels 1994), LR3R (www.biology.duke.edu/fungi/mycolab/primers.htm), and LR16 (Moncalvo et al. 1993), were used for sequencing to ensure good quality overlapping sequences were obtained. The PCR conditions, sequence alignment and subsequent phylogenetic analysis followed the methods of Crous et al. (2006a). The ITS1, ITS2 and 5.8S rRNA gene were only sequenced for isolates of which these data were not available. The ITS data were not included in the analyses but deposited in GenBank where applicable. Gaps longer than 10 bases were coded as single events for the phylogenetic analyses; the remaining gaps were treated as missing data. Sequence data were deposited in GenBank (Table 1) and alignments in TreeBASE (www.treebase.org).

Taxonomy
Structures were mounted in lactic acid, and 30 measurements (x 1 000 magnification) determined wherever possible, with the extremes of spore measurements given in parentheses. Colony colours (surface and reverse) were assessed after 2-4 wk on OA and PDA at 25 °C in the dark, using the colour charts of Rayner (1970). All cultures obtained in this study are maintained in the CBS collection (Table 1). Nomenclatural novelties and descriptions were deposited in MycoBank (www.MycoBank.org).

	RESULTS

TOP Abstract INTRODUCTION MATERIALS AND METHODS RESULTS Members of Venturiaceae Excluded taxa DISCUSSION References

 
DNA phylogeny
Amplicons of approximately 1 700 bases were obtained for the isolates listed in Table 1. These sequences were used to obtain additional sequences from GenBank which were added to the alignment. The manually adjusted LSU alignment contained 116 sequences (including the two outgroup sequences) and 1 157 characters including alignment gaps (available in TreeBASE). Of the 830 characters used in the phylogenetic analysis, 326 were parsimony-informative, 79 were variable and parsimony-uninformative, and 425 were constant. Neighbour-joining analyses using three substitution models on the sequence data yielded trees with identical topologies to one another. The neighbour-joining trees support the same clades as obtained from the parsimony analysis, but with a different arrangement at the deep nodes, for example, the clade containing Protoventuria alpina (Sacc.) M.E. Barr (CBS 140.83) is placed as sister to the Venturiaceae using parsimony but basal to the Herpotrichiellaceae using neighbour-joining. Because of the large number of different strain associations in the Venturia clade (see the small number of strict consensus branches for this clade in Fig. 1), only the first 5 000 equally most parsimonious trees (TL = 1 752 steps; CI = 0.392; RI = 0.849; RC = 0.333) were saved, one of which is shown in Fig. 1.

View larger version (78K): [in this window] [in a new window]   Fig. 1. One of 5 000 equally most parsimonious trees obtained from a heuristic search with 100 random taxon additions of the LSU sequence alignment using PAUP v. 4.0b10. The scale bar shows 10 changes, and bootstrap support values from 1 000 replicates are shown at the nodes. Thickened lines indicate the strict consensus branches and ex-type sequences are printed in bold face. The tree was rooted to two sequences obtained from GenBank (Athelia epiphylla AY586633 and Paullicorticium ansatum AY586693).

View larger version (79K): [in this window] [in a new window]   Fig. 2. Consensus phylogram (50 % majority rule) of 1 500 trees resulting from a Bayesian analysis of the LSU sequence alignment using MRBAYES v. 3.1.2. Bayesian posterior probabilities are indicated at the nodes. Ex-type sequences are printed in bold face. The tree was rooted to two sequences obtained from GenBank (Athelia epiphylla AY586633 and Paullicorticium ansatum AY586693).

Members of Chaetothyriales, Herpotrichiellaceae
Cladophialophora australiensis Crous & A.D. Hocking, sp. nov. MycoBank MB504525. Fig. 3.

View larger version (65K): [in this window] [in a new window]   Fig. 3. Cladophialophora australiensis (CBS 112793). A. Conidiophore. B-C. Subcylindrical ramoconidia, and ellipsoid conidia. Scale bar = 10 µm.

Cladophialophorae carrionii similis, sed conidiis secundis majoribus, (7-)8-12(-15) x 3-4 µm.

In vitro: Mycelium consisting of branched, septate, smooth, pale brown, guttulate, 2-3 µm wide hyphae; hyphal coils not seen. Conidiophores dimorphic; macroconidiophores mononematous, subcylindrical, multi-septate, straight to curved, up to 150 µm long (including conidiogenous cells), and 4 µm wide, pale to medium brown, smooth, guttulate; microconidiophores integrated with hyphae, which terminate in subcylindrical conidiogenous cells that give rise to branched chains of conidia; conidiophores (including conidiogenous cells) up to 5-septate, 50 µm long, with terminal and lateral conidiogenous cells. Conidiogenous cells pale to medium brown, smooth, guttulate, terminal and lateral, subcylindrical, 20-35 x 2-3.5 µm, or reduced to indistinct subtruncate to truncate loci, scars up to 2 µm wide, mono- to polyblastic, proliferating sympodially, scars neither darkened, thickened, nor refractive. Conidia pale to medium brown, guttulate, smooth; ramoconidia subcylindrical, 0-1-septate, 20-35 x 2-3 µm, hila subtruncate, inconspicuous, up to 2 µm wide, giving rise to branched chains of conidia; conidia ellipsoid, pale brown, but becoming dark brown and thick-walled in older cultures, guttulate, tapering towards subtruncate terminal loci, 0-1-septate, occurring in chains of up to 20 conidia, (7-)8-12(-15) x 3-4 µm (older, dark brown conidia are ellipsoid, up to 5 µm wide).

Cultural characteristics: Colonies erumpent, somewhat spreading, margins crenate, feathery, aerial mycelium sparse; colonies on PDA olivaceous-grey to iron-grey (surface); reverse iron-grey; on OA and SNA olivaceous-grey. Colonies reaching 5 mm diam after 2 wk at 25 °C in the dark; colonies fertile. Not able to grow at 37 °C.

Specimen examined: Australia, isolated from apple juice, Dec. 1986, A.D. Hocking, holotype CBS H-19899, culture ex-type CBS 112793 = CPC 1377.

Notes: Cladophialophora australiensis is one of two novel species of Cladophialophora originally isolated from sports drinks in Australia. Cladophialophora spp. are commonly associated with human disorders (Honbo et al. 1984, de Hoog et al. 2000, Levin et al. 2004), and thus their occurrence in sports drinks is cause for concern. However, none of the new species described here had the ability to grow at 37 °C, and therefore it is not expected that they could pose a danger to humans. Comparing ITS diversity, the species shows more than 12 % difference to established pathogens such as C. carrionii and C. bantiana (Sacc.) de Hoog et al.

Cladophialophora chaetospira (Grove) Crous & Arzanlou, comb. nov. MycoBank MB504526. Fig. 4.

View larger version (108K): [in this window] [in a new window]   Fig. 4. Cladophialophora chaetospira (CBS 114747). A-C. Hyphae giving rise to conidiophores with catenulate conidia. D-F. Conidia become up to 3-septate, frequently remaining attached in chains. Scale bars = 10 µm.

Septonema chaetospira (Grove) S. Hughes, Naturalist, London 840: 9. 1952.

Heteroconium chaetospira (Grove) M.B. Ellis, in Ellis, More Dematiaceous Hyphomycetes: 64. 1976.

In vitro: Mycelium consisting of branched, septate, smooth, medium brown hyphae, 2-3.5 µm wide. Conidiophores reduced to conidiogenous cells, or a single supporting cell, 20-40 x 3-4 µm. Conidiogenous cells subcylindrical, erect, straight to irregularly curved, medium brown, smooth, 15-30 x 3-4 µm. Conidia in branched, acropetal chains with up to 30 conidia; subcylindrical to fusiform, medium brown, smooth, tapering slightly at subtruncate ends, 1(-3)-septate, thin-walled, becoming slightly constricted at septa of older conidia, (20-)25-30(-45) x 3-4(-5) µm; conidia remaining attached in long chains; hila neither thickened, nor darkened-refractive.

Cultural characteristics: Colonies erumpent, convex, spreading, with sparse to dense aerial mycelium; margins smooth, undulate; on PDA iron-grey (surface), margins olivaceous-black; reverse olivaceous-black; on OA olivaceous-grey in the middle due to fluffy aerial mycelium, iron-grey in wide outer margin; on SNA olivaceous-grey. Colonies reaching 12 mm diam after 2 wk on PDA at 25 °C in the dark. Not able to grow at 37 °C.

Specimens examined: China, Yunnan, Yiliang, isolated from Phyllostachys bambusoides (Gramineae), decaying bamboo, freshwater, 6 Jul. 2003, L. Cai, CBS 114747; China, Yunnan, stream in Kunming, isolated from bamboo wood, 15 Jun. 2003, C. Lei, CBS 115468. Denmark, isolated from roots of Picea abies (Pinaceae), isol. by D.S. Malla, CBS 491.70. Germany, Schleswig-Holstein, Kiel-Kitzeberg, isolated from wheat field soil, isol. by W. Gams, CBS 514.63 = ATCC 16274 = MUCL 8310.

Notes: Two cultures of Heteroconium chaetospira were originally deposited as Spadicoides minuta L. Cai, McKenzie & K.D. Hyde (Cai et al. 2004), but later found to represent Heteroconium chaetospira, a species commonly found on rotting wood in Europe (Ellis 1976). The genus Heteroconium Petr. has in recent years been used to name leaf spotting fungi with chains of brown, disarticulating conidia (Crous et al. 2006b), which have phylogenetic affinities to several orders, obviously being polyphyletic. The type species of Heteroconium, H. citharexyli Petr., is a plant pathogen on Cytharexylum (Petrak 1949) with hitherto unknown phylogenetic position. The fact that H. chaetospira is linked to the Chaetothyriales, was rather unexpected. The species appears to be similar to others placed in Cladophialophora by having short, lateral conidiogenous cells, and long chains of branched subcylindrical conidia that largely remain attached. It is, however, quite distinct from other members of Cladophialophora in having medium brown conidia, and in lacking the ellipsoid conidia observed in several species.

Cladophialophora hostae Crous, U. Braun & H.D. Shin, sp. nov. MycoBank MB504527. Figs 5-6.

View larger version (112K): [in this window] [in a new window]   Fig. 5. Cladophialophora hostae (CPC 10737). A-B. Conidiogenous loci (arrows). C. Hyphal coil. D-F. Branched conidial chains. G-H. Conidia. Scale bar = 10 µm.

View larger version (21K): [in this window] [in a new window]   Fig. 6. Cladophialophora hostae (CPC 10737). Branched conidial chains with ramoconidia and conidia. Scale bar = 10 µm.

Cladophialophorae scillae similis, sed conidiophoris in vitro brevioribus et leniter angustioribus, 10-15 x 1.5-2 µm, conidiis brevioribus, (7-)10-15(-20) µm.

In vivo: Leaf spots amphigenous, subcircular to somewhat angular-irregular, 1-5 mm wide, scattered to aggregated, sometimes confluent, pale to medium brown or with a reddish brown tinge, later greyish brown, margin indefinite or on the upper leaf surface with a narrow slightly raised marginal line or very narrow lighter halo, yellowish, ochraceous to brownish. Caespituli epiphyllous, punctiform to confluent, dingy greyish brown. Mycelium immersed, forming fusicladium-like hyphal strands or plates; hyphae septate, sometimes with constrictions at the septa, thin-walled, pale olivaceous, 1.5-7 µm wide. Stromata immersed, small, 10-40 µm diam, composed of swollen hyphal cells, subcircular to somewhat angular-irregular in outline, 2-8 µm diam, wall somewhat thickened, brown. Conidiophores in small to moderately large fascicles, loose, divergent to moderately dense, rarely solitary, arising from stromatic hyphal aggregations, erumpent, erect, usually unbranched, rarely branched, straight, subcylindrical to distinctly geniculate-sinuous, 5-40 x 2-5 µm, 0-6-septate, pale to medium olivaceous to olivaceous-brown, thin-walled, up to 0.5 µm, smooth. Conidiogenous cells integrated, terminal, 5-15(-20) µm long, sympodial, conidiogenous loci rather inconspicuous to subdenticulate, flat-tipped, 1-1.5 µm diam, unthickened or almost so, not to slightly darkened-refractive. Conidia in simple or branched chains, narrowly ellipsoid-subcylindrical, 10-15 x 1.5-3.5 µm, 0-1-septate, subhyaline to pale olivaceous, thin-walled, smooth, ends truncate or with two denticle-like hila in ramoconidia, (0.75-)1-1.5(-2) µm diam, unthickened or almost so, at most slightly darkened-refractive.

In vitro: Mycelium composed of branched, smooth, pale olivaceous to medium brown hyphae, frequently forming hyphal coils, guttulate, septa inconspicuous, not constricted, hyphae somewhat irregular in width, 1-2 µm wide. Conidiophores reduced to conidiogenous cells, integrated in hyphae, terminal, subcylindrical, pale olivaceous to pale brown, smooth, 0-1-septate, proliferating sympodially at apex via 1-2(-3) flat-tipped, minute, denticle-like loci, 1-1.5 µm wide, 10-15 x 1.5-2 µm; scars minutely darkened and thickened, but not refractive. Conidia in extremely long chains (-60), simple or branched, subcylindrical, or narrowly ellipsoid, smooth, pale olivaceous, 0-1-septate, (7-)10-15(-20) x (1.5-)2(-2.5) µm, hila truncate, 1-1.5 µm wide, minutely thickened and darkened-refractive.

Cultural characteristics: Colonies on PDA erumpent, spreading, with smooth, undulate margins and dense aerial mycelium; surface hazel (middle), outer zone isabelline; reverse fuscous-black in middle, isabelline in outer zone. Colonies reaching 25 mm diam on SNA, and 40 mm diam on PDA after 1 mo at 25 °C in the dark; colonies fertile.

Specimen examined: Korea, Pyongchang, Hosta plantaginea (Hostaceae), 20 Sep. 2003, H.D. Shin, HAL 2030 F, holotype, culture ex-type SMK 19664, CPC 10737 = CBS 121637, CPC 10738-10739.

Notes: Although this species is morphologically similar to Cladophialophora scillae (Deighton) Crous, U. Braun & K. Schub. described below in this paper, C. hostae is treated as a separate taxon due to the differences in the length and width of its conidiophores and conidia in vitro, as well as 17 bp differences in the ITS DNA sequence data and a distinct ecology causing leaf-spots on a different, unrelated host. Based on disease symptoms caused on the living host leaves, C. hostae is a very unusual, unexpected member of the genus Cladophialophora. In vivo, the mycelium forms obvious hyphal strands and plates which are characteristic for Fusicladium species. The conidiophores and conidia are also fusicladium-like. Nevertheless, this species clusters within the Herpotrichiellaceae, i.e., it has to be placed in the genus Cladophialophora. Biotrophic species like C. hostae and C. scillae without phialidic synanamorphs render the differentiation between Cladophialophora and Fusicladium (incl. Pseudocladosporium) almost impossible without sequence data. Furthermore, the morphology of C. hostae in vivo and in vitro shows remarkable differences in conidiophore morphology, i.e., the growth in vivo is characteristically fusicladium-like (conidiophores macronematous, long, septate), whereas habit in vitro is rather pseudocladosporium-like (conidiophores less developed, usually reduced to conidiogenous cells, short). However, several Fusicladium species have also been observed to exibit a Pseudocladosporium growth habit in culture, suggesting this growth plasticity to be rather common, and strongly influenced by growth conditions.

Cladophialophora humicola Crous & U. Braun, sp. nov. MycoBank MB504528. Figs 7-8.

View larger version (122K): [in this window] [in a new window]   Fig. 7. Cladophialophora humicola (CBS 117536). Conidiophore with branched conidial chains. Scale bar = 10 µm.

View larger version (22K): [in this window] [in a new window]   Fig. 8. Cladophialophora humicola (CBS 117536). A. Hyphal coil. B. Conidiophore. C-F. Conidial chains with ramoconidia and conidia. Scale bar = 10 µm.

Cladophialophorae bantianae similis, sed conidiis majoribus, (8-)11-14(-17) x (1.5-)2(-2.5) µm, locis conidiogenis et hilis angustioribus, 1-1.5 µm latis.

In vitro: Mycelium composed of branched, smooth, pale olivaceous to pale brown hyphae, frequently forming hyphal coils, prominently guttulate, not to slightly constricted at the septa, 1-2 µm wide, cells somewhat uneven in width. Conidiophores solitary, mostly inconspicuous and integrated in hyphae, varying from inconspicuously truncate lateral loci on hyphal cells, 1-1.5 µm wide, to occasionally terminal conidiophores, 0-3-septate, subcylindrical, proliferating sympodially, 10-30 x 1.5-3 µm, pale brown, smooth.

Conidiogenous cells integrated, inconspicuous, truncate, lateral loci 1-1.5 µm wide, or conidiogenous cells subcylindrical with 1-3 sympodial loci (which appear as minute lateral denticles), 7-17 x 1.5-2 µm; scars inconspicuous, neither darkened, refractive nor thickened. Conidia in short chains of up to 10, simple or branched, subcylindrical to narrowly ellipsoid, 0-1-septate, (8-)11-14(-17) x (1.5-)2(-2.5) µm, pale olivaceous to olivaceous-brown or pale brown, smooth, hila truncate, 1-1.5 µm wide, unthickened, neither darkened, nor refractive.

Cultural characteristics: Colonies erumpent, spreading, with uneven, feathery margins and dense aerial mycelium on PDA; pale olivaceous-grey in the middle, becoming olivaceous-grey in the outer zone (surface); reverse olivaceous-black, with grey-olivaceous margins. Colonies reaching 7 mm diam after 2 wk at 25 °C in the dark; colonies fertile.

Specimen examined: Germany, Brandenburg, Müncheberg, from soil, Zaspel, Zalf & H. Nirenberg, holotype CBS H-19902, culture ex-type BBA 65570 = CBS 117536.

Notes: Phylogenetically Cladophialophora humicola is closely related to C. sylvestris Crous & de Hoog (see below). Morphologically the two species can be distinguished in that C. humicola lacks ramoconidia, and has 1-septate conidia, while those of C. sylvestris are 0-3-septate.

Cladophialophora potulentorum Crous & A.D. Hocking, sp. nov. MycoBank MB504529. Figs 9-10.

View larger version (131K): [in this window] [in a new window]   Fig. 9. Cladophialophora potulentorum (CBS 115144). A. Colony on PDA. B. Conidiophore. C-D. Conidial chains. E-F. Ramoconidia and conidia. Scale bar = 10 µm.

View larger version (22K): [in this window] [in a new window]   Fig. 10. Cladophialophora potulentorum (CBS 115144). Conidiophores with chains of ramoconidia and conidia. Scale bar = 10 µm.

Cladophialophorae carrionii similis, sed conidiis secundis majoribus, (6-)8-10(-13) x 2-3 µm.

In vitro: Mycelium consisting of branched, septate, smooth, pale brown, guttulate, 1.5-2.5 µm wide hyphae. Conidiophores solitary, macronematous, well distinguishable under the dissecting microscope from aerial mycelium, pale to medium brown, subcylindrical, straight to somewhat curved, erect, with apical apparatus appearing as a tuft due to extremely long conidial chains; conidiophores up to 5-septate, and 100 µm tall (excluding conidiogenous cells). Conidiogenous cells pale brown, smooth, terminal and lateral, subcylindrical, tapering towards subtruncate to truncate loci, 1 µm wide, somewhat darkened, thickened, but not refractive, loci appearing subdenticulate on lateral conidiogenous cells, mono- to polyblastic, proliferating sympodially, 10-35 x 1.5-2 µm. Conidia pale brown, smooth, guttulate, occurring in branched chains of up to 60; hila somewhat darkened and thickened, but not refractive, 0.5 µm wide; ramoconidia subcylindrical, 0-1-septate, 15-17(-20) x 2.5-3 µm; conidia ellipsoid, (6-)8-10(-13) x 2-3 µm.

Cultural characteristics: Colonies erumpent, spreading, with smooth margins and dense aerial mycelium on PDA, olivaceous-grey (surface), with a thin, olivaceous-black margin; reverse olivaceous-black; on OA olivaceous-grey (surface) with a wide olivaceous-black margin. Colonies reaching 25-30 mm diam after 1 mo at 25 °C in the dark; colonies fertile, also sporulating in the agar. Not able to grow at 37 °C.

Specimens examined: Australia, isolated from apple juice drink, Dec. 1986, A.D. Hocking, holotype CBS H-19901, culture ex-type CBS 115144 = CPC 11048; Australia, isolated from sports drink, Feb. 1996, A.D. Hocking, CBS 114772 = CPC 1375 = FRR 4947; Australia, isolated from sports drink, Feb. 1996, A.D. Hocking, CBS 112222 = FRR 4946.

Notes: Originally this taxon, isolated from fruit and sports drinks, was thought to be an undescribed species of Pseudocladosporium (= Fusicladium, see below). However, upon closer examination, this proved not to be the case. Conidiophores appear as distinct tufts under the dissecting microscope, and are readily distinguishable from the superficial mycelium, as is normally observed in species of Fusicladium, but the conidial chains are extremely long, and the conidia tend to be more ellipsoid than the predominantly fusiform or subcylindrical conidia observed in species of Fusicladium. Hyphal coils were also not observed in cultures of C. potulentorum, but are rather common in species of Fusicladium. The phylogenetic position of this taxon within the Herpotrichiellaceae clade also supports inclusion in the genus Cladophialophora.

View larger version (127K): [in this window] [in a new window]   Fig. 11. Cladophialophora proteae (CBS 111667). A. Colony on OA. B-C. Conidiophores. D-H. Catenulate conidia. Scale bars = 10 µm.

Pseudocladosporium proteae (Viljoen & Crous) Crous, in Crous et al., Cultivation and Diseases of Proteaceae: Leucadendron, Leucospermum and Protea: 101. 2004.

In vitro: Mycelium consisting of branched, septate hyphae, often forming strands, anastomosing, smooth to finely verruculose, frequently constricted at septa, olivaceous, 3-4 µm wide; hyphal cells in older cultures becoming swollen, up to 6 µm wide. Conidiophores reduced to conidiogenous cells. Conidiogenous cells holoblastic, integrated, forming short, truncate protuberances, 2-3 x 1.5-2 µm, concolorous with mycelium, subcylindrical. Conidia in vitro arranged in long acropetal chains (up to 20), simple or branched, subcylindrical to oblong-doliiform, (9-)13-17(-22) x 2.5-3(-4) µm in vitro on MEA, (9-)16-22(-25) x (2.5-)3-4(-6) µm on SNA; 0-1(-4)-septate, pale brown to pale olivaceous, smooth, hila subtruncate to truncate, not thickened, but somewhat refractive.

Cultural characteristics: Colonies erumpent, with sparse aerial mycelium on PDA; margins irregular, feathery; greyish rose, with patches of pale olivaceous-grey (surface); reverse olivaceous-grey. Colonies reaching 10 mm diam after 2 wk at 25 °C in the dark; colonies fertile.

Specimen examined: South Africa, Western Cape Province, Stellenbosch, J.S. Marais Nature Reserve, leaves of Protea cynaroides (Proteaceae), 26 Aug. 1996, L. Viljoen, holotype PREM 55345, culture ex-type CBS 111667.

Notes: Cladophialophora proteae differs from species of Fusicladium (= Pseudocladosporium) based on its colony colour, the slimy nature of colonies, as well as its conidia that have inconspicuous, unthickened hila (Fig. 11) (Crous et al. 2004), unlike those observed in species of Fusicladium. Sequence data show that this species is not allied to the Venturiaceae, but to the Herpotrichiellaceae.

Cladophialophora scillae (Deighton) Crous, U. Braun & K. Schub., comb. nov. MycoBank MB504530. Fig. 12.

View larger version (124K): [in this window] [in a new window]   Fig. 12. Cladophialophora scillae (CBS 116461). A-C. Conidiophores. D-F. Catenulate conidia. Scale bar = 10 µm.

Fusicladium scillae (Deighton) U. Braun & K. Schub., IMI Descriptions of Fungi and Bacteria 152: 1518. 2002.

In vivo: see Schubert & Braun (2002a) and Schubert et al. (2003).

In vitro: Mycelium consisting of branched, septate, smooth, green-brown to medium brown, guttulate hyphae, variable in width, 1.5-3 µm diam. Conidiophores lateral or terminal on hyphae, erect, straight to slightly flexuous, solitary, in some cases aggregated, subcylindrical, curved to geniculate-sinuous, unbranched, up to 55 µm long, 2-3 µm wide, 0-7-septate, septa in short succession, pale to medium brown, somewhat paler towards apices, smooth. Conidiogenous cells integrated, terminal or lateral as individual loci on hyphal cells, straight to curved, subcylindrical, up to 14(-18) µm long and 2 µm wide, pale to medium brown, smooth, with a single or few subdenticulate to denticulate loci at the apex due to sympodial proliferation, or reduced to individual loci, 0.8-1.5(-2) µm wide; scars minutely thickened and darkened, but not refractive. Conidia occurring in long, unbranched or loosely branched chains (-30), straight to slightly curved, ellipsoid to mostly narrowly subcylindrical, obclavate in some larger, septate conidia, (5-)10-20(-35) x 1.5-3 µm, 0-1(-3)-septate, sometimes slightly constricted at the septa, subhyaline to pale brown, smooth, guttulate, tapering at ends to subtruncate hila, 0.8-1.5 µm wide, minutely thickened and darkened, but not refractive; microcyclic conidiogenesis occurring.

Cultural characteristics: Colonies erumpent, spreading, with smooth, even margins and dense, abundant aerial mycelium on PDA; grey-olivaceous (surface); reverse dark olivaceous. Colonies on OA olivaceous-grey, smoke-grey due to profuse sporulation, reverse olivaceous-grey to iron-grey, velvety, aerial mycelium sparse, diffuse. Colonies reaching 20 mm diam on SNA, and 40 mm on PDA after 1 mo at 25 °C in the dark; colonies fertile.

Specimens examined: New Zealand, Levin, on Scilla peruviana (Hyacinthaceae), 21 Dec. 1965, G.F. Laudon, IMI 116997 holotype; Auckland, Manurewa, Auckland Botanic Gardens, on leaf spots of Scilla peruviana, 25 Apr. 2004, C.F. Hill, 1044, CBS H-19903, epitype designated here, culture ex-type CBS 116461.

Notes: In culture Cladophialophora scillae forms a pseudocladosporium-like state, though the scars are somewhat darkened and thickened, but not refractive. Conidiophores are reduced to conidiogenous cells that are integrated in the mycelium, terminal or lateral, frequently also as an inconspicuous lateral denticle, with a flat-tipped scar. Conidia occur in long, branched chains, which are subcylindrical to narrowly ellipsoid, and are up to 35 µm long, 1.5-3 µm wide, thus longer and thinner than reported on the host, which were 0-3-septate, subcylindrical to ellipsoid-ovoid, 7-22 x 2.5-4 µm. Due to the fusicladioid habit of this species in vivo, Schubert & Braun (2002a) reallocated it to Fusicladium. Based on ITS sequence data, morphology and cultural characteristics, Cladophialophora scillae was almost identical to an isolate obtained from leaf spots of Hosta plantaginea in Korea. These isolates appeared to resemble species of Fusicladium, but phylogenetically they clustered in the Herpotrichiellaceae. Therefore, "Fusicladium" scillae was placed in the genus Cladophialophora. As far as we are aware, this species and C. hostae are first reports of phytopathogenic species within the genus Cladophialophora.

Cladophialophora sylvestris Crous & de Hoog, sp. nov. MycoBank MB504531. Fig. 13.

View larger version (74K): [in this window] [in a new window]   Fig. 13. Cladophialophora sylvestris (CBS 350.83). A-B. Conidiophores. C. Catenulate conidia. D. Conidial mass. Scale bar = 10 µm.

View larger version (70K): [in this window] [in a new window]   Fig. 14. Cyphellophora hylomeconis (CBS 113311). A. Colony on PDA. B-C. Hyphae with truncate conidiogenous loci. D-F. Conidia. Scale bars = 10 µm.

Cladophialophorae humicolae similis, sed conidiis 0-3-septatis, (7-)10-16(-20) x 1.5-2 µm.

Mycelium composed of branched, smooth, pale olivaceous to pale brown hyphae, frequently forming hyphal coils, not to slightly constricted at the septa, 1-2 µm wide. Conidiophores medium brown, subcylindrical, flexuous, mononematous, multiseptate, up to 50 µm long, and 2-3 µm wide. Conidiogenous cells apical, sympodial, pale brown, 5-12 x 2-3 µm; scars somewhat darkened and thickened, not refractive. Conidia occurring in branched chains; ramoconidia up to 2 µm wide, giving rise apically to disarticulating chains of conidia; smooth, 0-3-septate, pale olivaceous, subcylindrical, (7-)10-16(-20) x 1.5-2 µm, with truncate ends; hila somewhat darkened and thickened, not refractive.

Cultural characteristics: Colonies erumpent on PDA, with smooth, catenulate margins; iron-grey (surface); reverse greenish black. Colonies reaching 15 mm diam after 1 mo at 25 °C in the dark; colonies fertile.

Specimen examined: Netherlands, Kootwijk, needle litter of Pinus sylvestris (Pinaceae), 8 Nov. 1982, G.S. de Hoog, holotype CBS H-19917, culture ex-type CBS 350.83.

Notes: Morphologically CBS 350.83 was originally identified as Polyscytalum griseum Sacc., but the latter is reported to have conidia that are 5-5.5 x 1 µm (Saccardo 1877), which is much smaller than that observed for the present isolate. Furthermore, the type species of Polyscytalum, P. fecundissimum Riess (CBS 100506), does not cluster within the Herpotrichiellaceae, thus suggesting that CBS 350.83 is best treated as a new species of Cladophialophora.

Cyphellophora hylomeconis Crous, de Hoog & H.D. Shin, sp. nov. MycoBank MB504532. Fig. 14.

Etymology: Named after its host genus, Hylomecon.

Cyphellophorae lacinatae similis, sed conidiis longioribus et leniter angustioribus, (15-)25-35(-55) x (2.5-)3(-4) µm.

Mycelium consisting of branched, greenish brown, septate, branched, smooth, 3-5 µm wide hyphae, constricted at septa. Conidiogenous cells phialidic, intercalary, appearing denticulate, 1 µm tall, 1.5-2 µm wide, with minute collarettes (at times proliferating percurrently). Conidia sickle-shaped, smooth, medium brown, guttulate, (1-)3(-5)-septate, constricted at septa, widest in middle, or lower third of the conidium; apex subacutely rounded, base subtruncate, or having a slight constriction, giving rise to a foot cell, 1 µm long, 0.5-1 µm wide, subacutely rounded, (15-)25-35(-55) x (2.5-)3(-4) µm; a marginal frill is visible above the foot cell, suggesting this foot cell may be the onset of basal germination; conidia also anastomose and undergo microcyclic conidiation in culture.

Cultural characteristics: Colonies slow-growing, slimy, aerial mycelium absent, margins smooth, catenate; surface crumpled, olivaceous-black to iron-grey. Colonies reaching 20 mm diam after 1 mo at 25 °C in the dark on PDA, 12 mm on SNA; colonies fertile.

Specimen examined: Korea, Yangpyeong, on leaves of Hylomecon verlance (Papaveraceae), 4 Jun. 2003, H.D. Shin, holotype CBS H-19907, isotype SMK 19550, culture ex-type CBS 113311.

Notes: Cyphellophora hylomeconis is related to the type species of the genus, Cyphellophora laciniata G.A. de Vries, which also resides in the Herpotrichiellaceae. The genus Cyphellophora G.A. de Vries is phenetically distinguished from Pseudomicrodochium B.C. Sutton, typified by P. aciculare B.C. Sutton (1975) by melanized versus hyaline thalli. Phylogenetic confirmation is pending due to unavailability of sequence data. Decock et al. (2003) synonymised the hyaline genus Kumbhamaya M. Jacob & D.J. Bhat (Jacob & Bhat 2000) with Cyphellophora, but as no cultures of this fungus are available this decision seems premature. Nearly all Cyphellophora species accepted by Decock et al. (2003) have been found to be involved in cutaneous infections in humans. This also holds true for the species originally described as being environmental, C. vermispora Walz & de Hoog, which is closely related to C. suttonii (Ajello et al.) Decock and C. fusarioides (C.K. Campbell & B.C. Sutton) Decock known from proven human and animal infections. Decock et al. (2003) added the melanized species C. guyanensis Decock & Delgado, isolated as a saprobe from tropical leaf litter. Cyphellophora hylomeconis is the first species of the genus infecting a living plant host. ITS sequences are remote from those of the remaining Cyphellophora species, the nearest neighbour being C. pluriseptata G.A. de Vries, Elders & Luykx at 19.1 % distance (data not shown). Cyphellophora hylomeconis can be distinguished based on its conidial dimensions and septation. Conidia are larger than those of C. fusarioides (11-20 x 2-2.5 µm, 1-2-septate), and those of C. laciniata (11-25 x 2-5 µm, 1-3-septate) (for a key to the species see Decock et al. 2003).

Exophiala sp. 1. Fig. 15.
Mycelium consisting of smooth, branched, septate, medium brown, 2-3 µm wide hyphae, regular in width, forming hyphal strands and hyphal coils, with free yeast-like cells present in culture; chlamydospores terminal on hyphae, frequently forming clusters or chains, medium brown, ellipsoid, 0-1-septate, up to 10 µm long and 5 µm wide. Conidiophores reduced to conidiogenous cells, or consisting of one supporting cell, giving rise to a single conidiogenous cell, subcylindrical to ellipsoid, medium brown, smooth, 5-12 x 3.5-4 µm, with 1(-3) phialidic loci, somewhat protruding, appearing subdenticulate at first glance under the light microscope. Conidiogenous cells integrated as lateral loci on hyphal cells, inconspicuous, 1-1.5 µm wide, with a slightly flaring collarette, (1-)1.5(-2) µm long. Conidia ellipsoid, smooth, guttulate, becoming brown, swollen and elongated, and at times 1-septate, 4-5(-7) x (2.5-)3(-4) µm (description based on CBS 115142).

View larger version (162K): [in this window] [in a new window]   Fig. 15. Exophiala sp. 1 (CBS 115142). A. Colony on PDA. B. Hyphal coil. C. Hyphal strand. D-H. Conidiogenous cells and loci. I-O. Conidiogenous cells and conidia. Scale bars = 10 µm.

Specimen examined: Australia, from a fruit drink, May 2002, N.J. Charley, CBS 115142 = CPC 11044 = FRR 5582.

Notes: Species of Exophiala are frequently observed as agents of human mycoses in immunocompromised patients (de Hoog et al. 2000). They are found in the environment as slow-growing, oligotrophic colonisers of moist substrates. For example the thermotolerant species E. dermatitidis (Kano) de Hoog and E. phaeomuriformis (Matsumoto et al.) Matos et al. are common in public steam baths (Matos et al. 2003), while E. mesophila Listemann & Freiesleben can be found in showers and swimming pools (unpubl. data). Both species are able to cause infections in humans (Zeng et al. 2007). Several other species have been associated primarily with infections in fish and cold-blooded animals (Richards et al. 1978) and are occasionally found on humans (Madan et al. 2006). The occurrence of the present species in fruit drinks, therefore, is cause of concern, although it was unable to grow at 37 °C. This species forms part of a larger study, and will be treated elsewhere.

Exophiala sp. 2. Fig. 16.
Mycelium consisting of smooth, branched, septate, pale brown, 1.5-3 µm wide hyphae, forming hyphal strands and hyphal coils; hyphae at times terminating in chains of ellipsoid chlamydospores that are medium brown, smooth, up to 10 µm long and 5 µm wide. Conidiophores subcylindrical, medium brown, smooth, consisting of a supporting cell and a single conidiogenous cell, or reduced to a conidiogenous cell, straight to curved, up to 30 µm long and 2-3 µm wide. Conidiogenous cells pale to medium brown, subcylindrical to narrowly ellipsoid or subclavate, with 1-3 apical, phialidic loci, 1 µm wide, 1-2 µm tall, collarette somewhat flaring, but mostly cylindrical, 7-20 x 2-2.5 µm; at times proliferating percurrently. Conidia ellipsoid, smooth, guttulate, hyaline, becoming pale olivaceous, apex obtuse, base subtruncate, (4-)5-7(-10) x 2-2.5(-3) µm.

View larger version (67K): [in this window] [in a new window]   Fig. 16. Exophiala sp. 2 (CBS 115143). A. Conidiogenous cells. B. Conidiophore with hyphal coil. C. Conidiogenous cell with hyphal strand. D. Conidia. Scale bar = 10 µm.

Specimen examined: Australia, from bottled spring water, May 2003, N.J. Charley, CBS 115143 = CPC 11047 = FRR 5599.

Notes: This strain represents another taxon occurring in bottled drinks destined for human consumption. As it is unable to grow at 37 °C, it does not appear to pose any serious threat to human health. This species forms part of a larger study, and will be treated elsewhere.

Exophiala eucalyptorum Crous, sp. nov. MycoBank MB504533. Fig. 17.

View larger version (132K): [in this window] [in a new window]   Fig. 17. Exophiala eucalyptorum (CPC 11261). A. Colony on PDA. B-H. Hyphae, conidiogenous cells and conidia. Scale bars = 10 µm.

Etymology: Named after its occurrence on Eucalyptus leaves.

Exophialae spiniferae similis, sed conidiis fusoidibus-ellipsoideis, (5-)6-8(-10) x (3-)4-5(-7) µm, et cellulis conidiogenis saepe catenatis, in catenis brevibus, dividentibus.

Mycelium consisting of smooth to finely verruculose, branched, septate, 2-4 µm wide hyphae, at times giving rise to chains of dark brown, fusoid-ellipsoid chlamydospores, which can still have phialides, suggesting they were conidiogenous cells; hyphae becoming constricted at septa when fertile. Conidiophores reduced to conidiogenous cells. Conidiogenous cells numerous, terminal and lateral, mono- to polyphialidic, 5-15 x 3-5 µm; loci 1-1.5 µm wide and tall, with inconspicuous collarettes, at time proliferating percurrently; conidiogenous cells fusoid-ellipsoid, and frequently breaking off, appearing as short chains of conidia, but distinct in having conidiogenous loci. Conidia fusoid-ellipsoid, apex acutely rounded, base subtruncate, (5-)6-8(-10) x (3-)4-5(-7) µm; frequently becoming fertile, septate and brown with age.

Cultural characteristics: Colonies erumpent, convex, smooth, slimy, margins feathery to crenate and smooth; aerial mycelium absent, growth yeast-like. Colonies on PDA, OA and SNA chestnut on surface and reverse. Colonies reaching 4 mm diam after 2 wk on PDA at 25 °C in the dark.

Specimen examined: New Zealand, Wellington Botanical Garden, on leaf litter of Eucalyptus sp. (Myrtaceae), Mar. 2004, J.A. Stalpers, holotype CBS H-19905, culture ex-type CBS 121638 = CPC 11261.

Notes: Exophiala eucalyptorum is rather characteristic in that, in culture, chains of conidiogenous cells frequently detach from hyphae, appearing as short, intact chains of fertile conidia. Its phylogenetic position is somewhat outside the core of the Herpotrichiellaceae containing most Capronia teleomorphs and the remaining opportunistic Exophiala species, but still within the Chaetothyriales (Figs 1-2).

	Members of Venturiaceae

TOP Abstract INTRODUCTION MATERIALS AND METHODS RESULTS Members of Venturiaceae Excluded taxa DISCUSSION References

 
Anungitea B. Sutton and Anungitopsis R.F. Castañeda & W.B. Kendr.
Sutton (1973) erected the genus Anungitea to accommodate species with brown, mononematous conidiophores bearing apically aggregated, flat-tipped, subdenticulate conidiogenous loci that give rise to chains of pale brown subcylindrical conidia with thickened, darkened hila. He compared the type species, A. fragilis B. Sutton with anamorph genera of the Mycosphaerellaceae, but did not compare it to Fusicladium, to which it is remarkably similar. Castañeda & Kendrick (1990b) introduced the genus Anungitopsis based on A. speciosa R.F. Castañeda & W.B. Kendr. This genus was distinguished from Anungitea by its formation of subdenticulate conidiogenous loci distributed along the apical region of the conidiophore, and by the relatively poorly defined appearance of these loci. No cultures are available of the ex-type species of Anungitea, but we studied strains of Anungitopsis amoena R.F. Castañeda & Dugan (CBS 254.95, ex-type), and Anungitopsis intermedia Crous & W.B. Kendr. (CBS 110746, ex-epitype), and found them to cluster adjacent to Fusicladium (Venturiaceae). However, the ex-type strain of Anungitopsis speciosa (CBS 181.95), type species of Anungitopsis, clustered distantly from all other species, confirming that the genus name Anungitopsis is not available for any of the taxa treated here. In any case, A. speciosa has unusual subdenticulate conidiogenous loci with indistinct marginal frills, and these are obviously different from those of anungitea- and fusicladium-like anamorphs, including A. amoena and A. intermedia. The latter two species previously referred to as Anungitopsis belong to a sister clade of the Venturia (Fusicladium, incl. Pseudocladosporium) clade. Sympoventuria (Crous et al. 2007b), which produces a sympodiella-like anamorph in culture, is the only teleomorph of this clade hitherto known. The venturia-like habit of Sympoventuria, connected with fusicladium- / pseudocladosporium-like anamorphs distributed in both clades, indicates a close relation between these clades, suggesting a placement in the Venturiaceae. Schubert et al. (2003) referred to the difficulty to distinguish between Anungitea and Fusicladium. Anungitea is undoubtedly heterogeneous. Anungitea rhabdospora P.M. Kirk (Kirk 1983) is, for instance, intermediate between Anungitea (conidiophores with a terminal denticulate conidiogenous cell, but conidia disarticulating in an arthroconidium-like manner) and Sympodiella B. Kendr. (conidiophores distinctly sympodial, forming arthroconidia). Other species assigned to Anungitea possess a distinctly swollen, lobed conidiophore base, e.g. A. heterospora P.M. Kirk (Kirk 1983), which is comparable with other morphologically similar genera, e.g., Parapleurotheciopsis P.M. Kirk (Kirk 1982), Rhizocladosporium Crous & U. Braun (see Crous et al. 2007a - this volume), and Subramaniomyces Varghese & V.G. Rao (Varghese & Rao 1979, Kirk 1982). The application of Anungitea depends, however, on the affinity of A. fragilis, the type species, of which sequence data are not yet available. The best solution for this problem is the widened application of Fusicladium (incl. Pseudocladosporium) to both sister clades, i.e., to the whole Venturiaceae. Morphologically a distinction between fusicladioid anamorphs of both clades is impossible. The more "fusicladium-like" growth is mainly characteristic for the fruiting in vivo, above all in biotrophic taxa, whereas the more "pseudocladosporium-like" habit is typical for the growth in vitro and in saprobic taxa, a phenomenon which is also evident in species of the morphologically similar genus Cladophialophora (see C. hostae and C. scillae). A potential placement of Anungitea fragilis within the Venturiaceae, which has still to be proven, would render the genus Anungitea a synonym of Fusicladium, but in the case of a quite distinct phylogenetic position a new circumscription of this genus, excluding the Venturiaceae anamorphs, would be necessary. Thus, a final conclusion about Anungitea has to be postponed, awaiting cultures and sequence analyses of its type species.

View larger version (61K): [in this window] [in a new window]   Fig. 18. Cylindrosympodium lauri (CBS 240.95). A-C. Conidiophores with conidiogenous loci. D. Conidia. Scale bar = 10 µm.

Cylindrosympodium lauri Crous & R.F. Castañeda, sp. nov. MycoBank MB504534. Fig. 18.

Etymology: Named after the host genus it was collected from, Laurus.

Cylindrosympodii variabilis similis, sed conidiophoris longioribus, ad 70 µm, conidiis subhyalinis vel dilute olivaceis.

Mycelium consisting of brown, smooth, septate, branched hyphae, 1.5-2.5 µm wide. Conidiophores macronematous, mononematous, solitary, erect, subcylindrical, straight to geniculate-sinuous, medium brown, smooth, 35-70 x 2.5-4 µm, 1-5-septate. Conidiogenous cells terminal, integrated, pale to medium brown, smooth, 10-35 x 2-3 µm, proliferating sympodially, with one to several flat-tipped loci, 1.5-2 µm wide; scars somewhat darkened, minutely thickened, but not refractive. Conidia solitary, subacicular to narrowly subcylindrical, apex subobtuse, base truncate, or somewhat swollen, straight or curved, smooth, subhyaline to very pale olivaceous, guttulate, (45-)60-70(-80) x 2.5-3(-3.5) µm, (4-)6-8-septate; scars are somewhat darkened, minutely thickened, but not refractive, 2.5-3 µm wide.

Cultural characteristics: Colonies erumpent, convex, with smooth, lobed margins, and moderate, dense aerial mycelium on PDA; mouse-grey in the central part, and dark mouse-grey in the outer zone (surface); reverse dark mouse-grey. Colonies reaching 5 mm diam after 2 wk at 25 °C in the dark; colonies fertile.

Specimen examined: Spain, Canary Islands, leaf litter of Laurus sp. (Lauraceae), 4 Jan. 1995, R.F. Castañeda, holotype CBS H-19909, culture ex-type CBS 240.95.

Note: The present fungus differs from Cylindrosympodium variabile (de Hoog) W.B. Kendr. & R.F. Castañeda (de Hoog 1985) in that the conidiophores are much longer, the conidia are subhyaline to very pale olivaceous, and the scars and hila are thin, slightly darkened, but not refractive.

Venturia Sacc. and its anamorph Fusicladium
Venturia Sacc., Syll. fung. (Abellini) 1: 586. 1882.

= Apiosporina Höhn., Sitzungsber. Kaiserl. Akad. Wiss., Math.-Naturw. Cl., Abt. 1, 119: 439. 1910, syn. nov.

= Metacoleroa Petr., Ann. Mycol. 25: 332. 1927, syn. nov.

= Caproventuria U. Braun, A Monograph of Cercosporella, Ramularia and Allied Genera (Phytopathogenic Hyphomycetes) 2: 396. 1998, syn. nov.

For additional synonyms see Sivanesan, The bitunicate Ascomycetes and their anamorphs: 604. 1984.

Anamorph: Fusicladium Bonord., Handb. Mykol.: 80. 1851.

= Pseudocladosporium U. Braun, A Monograph of Cercosporella, Ramularia and Allied Genera (Phytopathogenic Hyphomycetes) 2: 392. 1998, syn. nov.

For additional synonyms, see Schubert et al. (2003).

Notes: The genus Caproventuria, based on C. hanliniana (U. Braun & Feiler) U. Braun, was erected to accommodate saprobic, soil-borne venturia-like ascomycetes with numerous ascomatal setae, and an anamorph quite distinct from Fusicladium (Braun 1998). The genus Metacoleroa is based on M. dickiei (Berk. & Broome) Petr., which clusters in the Venturiaceae, adjacent to Caproventuria, which has Pseudocladosporium anamorphs. Metacoleroa was retained by Barr (1987) as separate from Venturia based on its superficial ascomata with a thin, stromatic layer beneath the ascomata. Whether these criteria still justify the separation of Caproventuria and Metacoleroa from Venturia is debatable, and the names Venturia dickiei (Berk. & Broome) Ces. & de Not. and Venturia hanliniana (U. Braun & Feiler) Unter. are available for these organisms. The genus Apiosporina, which is based on Apiosporina collinsii (Schwein.) Höhn., clusters in the Venturiaceae, as was to be expected based on its Fusicladium anamorph (Schubert et al. 2003). It was distinguished from Venturia species by having ascospores strictly septate near the lower end (Sivanesan 1984).

The anamorph genus Fusicladium has been monographed by Schubert et al. (2003). Morphological as well as molecular studies (Beck et al. 2005) demonstrated that the genus Venturia with its Fusicladium anamorphs is monophyletic. A separation of Venturia into various uniform subclades based on the previous anamorph genera Fusicladium, Pollaccia and Spilocaea was not evident and could be rejected. As in cercosporoid anamorphs of Mycosphaerella, features such as the arrangement of the conidiophores (solitary, fasciculate, sporodochial), the proliferation of conidiogenous cells (sympodial, percurrent) and shape, size as well as formation of conidia (solitary, catenate) proved to be of little taxonomic value at generic level. Hence, Schubert et al. (2003) proposed to maintain Fusicladium emend. as sole anamorph genus for Venturia. The genus Fusicladosporium Partridge & Morgan-Jones (type species: Cladosporium carpophilum Thüm.) (Partridge & Morgan-Jones 2003), recently erected to accommodate fusicladium-like species with catenate conidia, represents a further synonym of Fusicladium.

Similar to their occurrence in vivo the conidiophores in vitro of species previously referred to the genera Spilocaea and Pollaccia are usually micronematous, conidia often appear to be directly formed on the mycelium, unilocal, determinate, mostly reduced to conidiogenous cells, sometimes forming a few percurrent proliferations, whereas the conidiophores of species of Fusicladium s. str. are mostly macronematous, but sometimes also micronematous. They are often initiated as short lateral, peg-like outgrowths of hyphae which proliferate sympodially, becoming slightly geniculate, forming a single, several or numerous subdenticulate to denticulate, truncate, unthickened or only slightly thickened, somewhat darkened-refractive conidiogenous loci.

The genus Pseudocladosporium was described to be quite distinct from Fusicladium by being saprobic and connected with a different teleomorph, viz. Caproventuria (Braun 1998). However, since th