Phylogeny and taxonomy of obscure genera of microfungi.

The recently generated molecular phylogeny for the kingdom Fungi, on which a new classification scheme is based, still suffers from an under representation of numerous apparently asexual genera of microfungi. In an attempt to populate the Fungal Tree of Life, fresh samples of 10 obscure genera of hyphomycetes were collected. These fungi were subsequently established in culture, and subjected to DNA sequence analysis of the ITS and LSU nrRNA genes to resolve species and generic questions related to these obscure genera. Brycekendrickomyces (Herpotrichiellaceae) is introduced as a new genus similar to, but distinct from Haplographium and Lauriomyces. Chalastospora is shown to be a genus in the Pleosporales, with two new species, C. ellipsoidea and C. obclavata, to which Alternaria malorum is added as an additional taxon under its oldest epithet, C. gossypii. Cyphellophora eugeniae is newly described in Cyphellophora (Herpotrichiellaceae), and distinguished from other taxa in the genus. Dictyosporium is placed in the Pleosporales, with one new species, D. streliziae. The genus Edenia, which was recently introduced for a sterile endophytic fungus isolated in Mexico, is shown to be a hyphomycete (Pleosporales) forming a pyronellea-like synanamorph in culture. Thedgonia is shown not to represent an anamorph of Mycosphaerella, but to belong to the Helotiales. Trochophora, however, clustered basal to the Pseudocercospora complex in the Mycosphaerellaceae, as did Verrucisporota. Vonarxia, a rather forgotten genus of hyphomycetes, is shown to belong to the Herpotrichiellaceae and Xenostigmina is confirmed as synanamorph of Mycopappus, and is shown to be allied to Seifertia in the Pleosporales. Dichotomous keys are provided for species in the various genera treated. Furthermore, several families are shown to be polyphyletic within some orders, especially in the Capnodiales, Chaetothyriales and Pleosporales.

INTRODUCTION

The recent ‘Deep Hypha’ issue of Mycologia (vol. 98, 2006) included 21 phylogenetic studies employing multi-gene phylogenies to resolve major groups of Fungi. These papers provided the foundation for the study of James et al. (2006), in which six genes (SSU, LSU, 5.8S rRNA, rpb1, rpb2 and tef1) for approximately 200 fungal taxa were used to present the first kingdom-level phylogeny, and a new classification for the Fungi (Hibbett et al. 2007). These studies also illustrated clearly that it was merely the ‘tip of the iceberg’, and that numerous genera must now be accommodated in this phylogenetic framework.

A major problem encountered during the Assembling the Fungal Tree of Life (AFTOL, www.aftol.org) project, was that many genera are insufficiently known, and have never been cultured, or subjected to DNA analyses. This is especially true for the majority of apparently asexual microfungi, namely the coelomycetes (Sutton 1980, Nag Raj 1993) and hyphomycetes (Ellis 1971, 1976, Carmichael et al. 1980). The only means to deal with this problem is, therefore, to encourage mycologists to recollect these genera and species, to establish cultures for them and to ultimately generate DNA sequence data (Shenoy et al. 2007), a process which can be described as ‘leafing out the fungal tree of life’.

Ten genera of hyphomycetes not previously known from culture, or for which the phylogenetic classification is uncertain, are treated in the present study. These fungi were collected from diverse hosts from various continents, isolated in axenic culture, and subjected to DNA sequence analysis. They are shown to belong to the Chaetothyriales (Brycekendrickomyces, Cyphellophora, Vonarxia), Pleosporales (Chalastospora, Dictyosporium, Edenia, Xenostigmina), Helotiales (Thedgonia), and the Capnodiales, Mycosphaerellaceae (Trochophora, Verrucisporota).

The present paper represents a further contribution in a series aiming to clarify the morphology and DNA phylogeny of obscure genera of microfungi. Other than resolving their phylogenetic relationships, several novelties are described, and keys are provided to the accepted species in these genera.

MATERIAL AND METHODS

Isolates

Symptomatic leaves and leaf litter were collected on various continents, and sent to the Centraalbureau voor Schimmelcultures (CBS) for isolation of microfungi. Leaves with visible fruiting were immediately subjected to direct isolation of hyphomycetes, or alternatively were first incubated in moist chambers to stimulate sporulation. Single-conidial isolates were established on malt extract agar (MEA; 20 g/L Biolab malt extract, 15 g/L Biolab agar) using the technique outlined in Crous (1998). Cultures were later plated on fresh MEA, 2 % water agar (WA) supplemented with sterile pine needles, 2 % potato-dextrose agar (PDA), synthetic nutrient agar (SNA) and/or oatmeal agar (OA) (Crous et al. 2009), and subsequently incubated at 25 °C under near-ultraviolet light to promote sporulation. Reference strains are maintained in the culture collection of the CBS, Utrecht, the Netherlands (Table 1). Descriptions, nomenclature, and illustrations were deposited in MycoBank (www.mycobank.org, Crous et al. 2004b).

Table 1

Collection details and GenBank accession numbers for fungal species included in this study.

Species	Strain no.1	Substrate	Country	Collector(s)	GenBank Accession no.2
					ITS	LSU
Brycekendrickomyces acaciae	CBS 124104; CPC 15078	Acacia auriculiformis	Indonesia	M.J. Wingfield	FJ839606	FJ839641
Chalastospora cetera	CBS 121340; E.G.S. 41.072	Elymus scabrus	Australia	R.G. Rees	FJ839607	FJ839642
Chalastospora ellipsoidea	CBS 121331; E.G.S. 22.060	Triticum sp.	Australia	H.L. Harvey & S. Perth	FJ839608	FJ839643
Chalastospora gossypii	CBS 112844; CPC 4571	Bromus tectorum	USA	F.M. Dugan	AY251081	AY251081
	CBS 114005; CPC 4572	Festuca idahoensis	USA	F.M. Dugan	AY251079	AY251079
	CBS 114809; MAF 943	Leaves of Anethum graveolens (dill) along with Itersonilia perplexans	New Zealand	J. Pike	FJ839609	FJ839644
	CBS 114810; MAF 954	Quercus robur (oak) leaves in association with Tubakia dryina	New Zealand	H. Nettleton	FJ839610	FJ839645
	CBS 148.66; CPC 3690; NRRL W 52-29	—	USA	C.W. Hesseltine	FJ839611	FJ839646
	CBS 173.80; ATCC 200939; CPC 3685	Agricultural soil	Syria	M.I.A. Abdel-Kader	—	FJ839647
	CBS 216.65; NRRL A-13702	Triticum aestivum grain	USA	C.W. Hesseltine	FJ839612	DQ008142
	CBS 266.75; ATCC 28332; CPC 3680; IMI 165252; PRE 44703	Wheat stubble	South Africa	W.F.O. Marasas	FJ839613	FJ839648
	CBS 900.87; ATCC 200938	Soil	Lebanon	F. Seigle-Murandi	FJ839614	FJ839649
	CPC 15567; C.F. Hill 2008/3899	Wood and wallpaper from inside walls of a dwelling	New Zealand	D. De Vanny	FJ839615	FJ839650
Chalastospora gossypii var. polymorpha	CBS 112048; CPC 4570	Dormant buds (overwintered) of Vitis vinifera	USA	F.M. Dugan	AY251080	AY251080
Chalastospora obclavata	CBS 124120; E.G.S. 12.128	Air	USA	C.T. Rogerson	FJ839616	FJ839651
Cyphellophora eugeniae	CBS 124105; CPC 15172	Living leaves of Stenocalyx uniflorus	Brazil	A.C. Alfenas	FJ839617	FJ839652
Dictyosporium strelitziae	CBS 123359; CPC 15359	Dead leaves of Strelitzia nicolai	South Africa	A. Wood	FJ839618	FJ839653
Edenia gomezpompae	CBS 124106; CPC 15689	Senna alata	Phillippines	C.J.R. Cumagun	FJ839619	FJ839654
Haplographium catenatum	CBS 196.73	Decaying wood	Germany	W. Gams	FJ839620	FJ839655
	CBS 482.67; CMW 754	Decaying wood	Germany	W. Gams	FJ839621	FJ839656
	CBS 739.68; CMW 755	Decaying wood	Netherlands	H.A. van der Aa	FJ839622	FJ839657
Lauriomyces bellulus	CBS 517.93	Cupule of Castanea sativa	Switzerland	P.W. Crous	FJ839623	FJ839658
Lauriomyces heliocephalus	CBS 112054; INIFAT CO2/59	Decaying leaf	Brazil	A. Stchigel & J. Guarro	FJ839624	FJ839659
Mycopappus aceris	CBS 124109; CPC 14379	Fallen leaves of Acer macrophyllum	Canada	B. Callan	FJ839625	FJ839660
Mycosphaerella lupini	CPC 1661	Lupinus sp.	USA	W. Kaiser	AF362050	FJ839661
Stenella anthuriicola	CBS 118742	Anthurium leaf	Thailand	C. F. Hill	FJ839626	FJ839662
Stigmina platani	CBS 110755; CPC 4299; IMI 136770	Platanus orientalis	India	—	AY260090	FJ839663
Thedgonia ligustrina	CPC 10019	Ligustrum ovalifolium	South Korea	H.-D. Shin	FJ839627	FJ839664
	CPC 10530	Ligustrum ovalifolium	Netherlands	P.W. Crous	FJ839628	FJ839665
	CPC 10861	Ligustrum ovalifolium	South Korea	H.-D. Shin	FJ839629	FJ839666
	CPC 14754	Ligustrum obtusifolium	South Korea	H.-D. Shin	FJ839630	FJ839667
	CPC 4298; W1877	Ligustrum sp.	Asia	H. Evans	EU040242	EU040242
Trochophora fasciculata	CPC 10281	Leaves of Daphniphyllum macropodum	South Korea	H.-D. Shin	FJ839631	—
	CPC 10282	Leaves of Daphniphyllum macropodum	South Korea	H.-D. Shin	FJ839632	FJ839668
Verrucisporota daviesiae	CBS 116002; VPRI 31767	Living leaves of Daviesia mimosoides	Australia	V. & R. Beilharz	FJ839633	FJ839669
Verrucisporota grevilleae	CBS 124107; CPC 14761	Leaves of Grevillia decurrens	Australia	B. Summerell	FJ839634	FJ839670
Verrucisporota proteacearum	CBS 116003; VPRI 31812	Grevillea sp.	Australia	V. Beilharz	FJ839635	FJ839671
Vonarxia vagans	CBS 123533; CPC 15151	Stenocalyx uniflorus	Brazil	A.C. Alfenas	FJ839636	FJ839672
	CPC 15152	Stenocalyx uniflorus	Brazil	A.C. Alfenas	FJ839637	FJ839673
Xenostigmina zilleri	CBS 115685; CPC 4011	Living leaves of Acer sp.	Canada	K.A. Seifert	FJ839638	FJ839674
	CBS 124108; CPC 14376	Fallen leaves of Acer macrophyllum	Canada	B. Callan	FJ839639	FJ839675
	CBS 115686; CPC 4010	Living leaves of Acer sp.	Canada	K.A. Seifert	FJ839640	FJ839676

1ATCC: American Type Culture Collection, Virginia, USA; C.F. Hill: Culture collection of C.F. Hill, housed at MAF, New Zealand; CBS: CBS Fungal Biodiversity Centre, Utrecht, The Netherlands; CMW: Culture collection of M.J. Wingfield, housed at FABI, Pretoria, South Africa; CPC: Culture collection of P.W. Crous, housed at CBS; E.G.S.: Culture collection of E.G. Simmons, Indiana USA; IMI: International Mycological Institute, CABI-Bioscience, Egham, Bakeham Lane, U.K.; INIFAT: Alexander Humboldt Institute for Basic Research in Tropical Agriculture, Ciudad de La Habana, Cuba; MAF: Ministry of Agriculture and Forestry, New Zealand; NRRL: National Center for Agricultural Utilization Research, Peoria, USA; PRE: National collection of fungi, Pretoria, South Africa; VPRI: Victorian Department of Primary Industries, Knoxfield, Australia.

2ITS: Internal transcribed spacers 1 and 2 together with 5.8S nrDNA; LSU: 28S nrDNA.

DNA isolation, amplification and analyses

Genomic DNA was isolated from fungal mycelium grown on MEA, using the UltraCleanTM Microbial DNA Isolation Kit (Mo Bio Laboratories, Inc., Solana Beach, CA, USA) according to the manufacturer’s protocols. The Primers V9G (de Hoog & Gerrits van den Ende 1998) and LR5 (Vilgalys & Hester 1990) were used to amplify part 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 (ITS2) and the first 900 bases at the 5’ end of the 28S rRNA gene (LSU). The primers ITS4 (White et al. 1990) and LR0R (Rehner & Samuels 1994) were used as internal sequence primers to ensure good quality sequences over the entire length of the amplicon. The PCR conditions, sequence alignment and subsequent phylogenetic analysis followed the methods of Crous et al. (2006b). Alignment gaps were treated as new character states. Sequence data were deposited in GenBank (Table 1) and alignments in TreeBASE (www.treebase.org). The ITS sequences were compared with those sequences available in NCBI’s GenBank nucleotide database using a megablast search and the results are discussed where applicable under the taxonomic notes. Because the genus Chalastospora is relatively novel, species in this genus were supported by a separate phylogenetic tree.

Morphology

Fungal descriptions were based on cultures sporulating in vitro (media indicated). Wherever possible, 30 measurements (× 1 000 magnification) were made of structures mounted in lactic acid, with the extremes of spore measurements given in parentheses. Colony colours (surface and reverse) were assessed after 2–4 wk on different media at 25 °C in the dark, using the colour charts of Rayner (1970).

RESULTS

Phylogenetic analysis

Amplification products of approximately 1 700 bases were obtained for the isolates listed in Table 1. The LSU region of the sequences was used to obtain additional sequences from GenBank, which were added to the alignment. Due to the inclusion of the shorter LSU sequences of Dictyosporium alatum (GenBank accession DQ018101), Dictyosporium elegans (GenBank accession DQ018100) and Dictyosporium toruloides (GenBank accession DQ018104) in the alignment, it was not possible to subject the full length of the determined LSU sequences (Table 1) to analyses. The manually adjusted LSU alignment contained 115 sequences (including the two outgroup sequences) and, of the 568 characters used in the phylogenetic analyses, 267 were parsimony informative, 30 were variable and parsimony uninformative, and 271 were constant. Neighbour-joining analyses using three substitution models on the sequence data yielded trees supporting the same tree topology to one another but differed from the most parsimonious tree shown in Fig. 1 with regard to the placement of the clade containing Ochroconis and Fusicladium (in the distance analyses, this clade moves to a more basal position). Forty equally most parsimonious trees (TL = 1 039 steps; CI = 0.477; RI = 0.833; RC = 0.397), the first of which is shown in Fig. 1, were obtained from the parsimony analysis of the LSU alignment.

Fig. 1

The first of 1 000 equally most parsimonious trees obtained from a heuristic search with 100 random taxon additions of the LSU sequence alignment. The scale bar shows 10 changes, and bootstrap support values from 1 000 replicates are shown at the nodes. Novel sequences generated for this study are shown in bold. Branches present in the strict consensus tree are thickened. Orders and families are coded as indicated in the legends. The tree was rooted to a sequence of Kluyveromyces lodderae (GenBank accession AY048161) and Saccharomyces cerevisiae (GenBank accession Z73326).

Abbreviations used: Families: AMO = Amorphothecaceae, CHA = Chaetothyriaceae, HEL = Helotiaceae, HER = Herpotrichiellaceae, HYA = Hyaloscyphaceae, IC = Incertae cedis: LEP = Leptosphaeriaceae, LOP = Lophiostomataceae, MEL = Melanommataceae, MYC = Mycosphaerellaceae, PHA = Phaeosphaeriaceae, PLE = Pleosporaceae, PSE = Pseudeurotiaceae, RHY = Rhytismataceae, SCH = Schizothyriaceae, TER = Teratosphaeriaceae. Orders: 1 = Capnodiales, 2 = Chaetothyriales, 3 = Incertae cedis, 4 = Rhytismatales, 5 = Helotiales, 6 = Hypocreales, 7 = Pezizales, 8 = Pleosporales.

The manually adjusted ITS alignment contained 28 sequences (including the outgroup sequence) and, of the 521 characters used in the phylogenetic analyses, 97 were parsimony informative, 91 were variable and parsimony uninformative, and 333 were constant. Neighbour-joining analyses using three substitution models on the sequence data yielded trees supporting the same tree topology to one another but differed from the most parsimonious tree shown in Fig. 2 with regard to the placement of Chalastospora ellipsoidea (in the distance analyses, this taxon moves to a more basal position in Chalastospora). Six equally most parsimonious trees (TL = 253 steps; CI = 0.913; RI = 0.938; RC = 0.856), the first of which is shown in Fig. 2, were obtained from the parsimony analysis of the ITS alignment. The results of the phylogenetic analyses are highlighted below under the taxonomic notes, or in the Discussion, where applicable.

Fig. 2

The first of 6 equally most parsimonious trees obtained from a heuristic search with 100 random taxon additions of the ITS sequence alignment. The scale bar shows 10 changes, and bootstrap support values (blue are from the parsimony analysis and red from the distance analysis using the HKY85 substitution model) from 1 000 replicates are shown at the nodes. Branches present in the strict consensus tree are thickened. The tree was rooted to a sequence of Phaeosphaeriopsis musae (GenBank accession DQ885894).

Taxonomy

Crous & M.J. Wingf., gen. nov. — MycoBank MB509515

Mycelium ex hyphis ramosis, septatis, laevibus, pallide brunneis, 1–2 μm latis compositum. Conidiophora solitaria, erecta, cylindrica, recta vel leviter flexuosa, cellula basali bulbosa, sine rhizoideis, stipite modice brunneo vel atro-brunneo, laevi, transverse euseptato, ad apicem cum (1–)2–4(–6) cellulis conidiogenis. Cellulae conidiogenae subcylindricae, allontoides vel doliiformes, rectae vel leviter curvatae, pallide brunneae, polyblasticae, sympodialiter proliferantes. Conidia hyalina, mucilagine aggregata (sed non catenata), ellipsoidea, apice subobtuso, basi subtruncata.

Type species. Brycekendrickomyces acaciae Crous & M.J. Wingf.

Etymology. Named for Bryce Kendrick, husband of Laurie Kendrick, for which Lauriomyces was named and that resembles the current genus.

Mycelium consisting of branched, septate, smooth, pale brown, 1–2 μm wide hyphae. Conidiophores solitary, erect, cylindrical, straight to somewhat flexuous, basal cell bulbous, without rhizoids; stalk medium to dark brown, smooth, transversely euseptate; upper cell giving rise to (1–)2–4(–6) conidiogenous cells. Conidiogenous cells subcylindrical to allantoid or doliiform, straight to gently curved, pale brown, polyblastic, proliferating sympodially. Conidia hyaline, aggregating in slimy mass (never in chains), ellipsoid, apex subobtuse, base subtruncate.

Crous & M.J. Wingf., sp. nov. — MycoBank MB509517; Fig. 3

Fig. 3

Brycekendrickomyces acaciae (CBS 124104). a. Colonies sporulating on MEA; b. colonies on SNA; c–e, g. conidiophores with conidiogenous apparatus; f. conidia. — Scale bars = 10 μm.

Maculae modice brunneae vel atro-brunneae, margine elevato, rubro-purpureo, oblongae vel ellipticae, ad 7 mm diam, in consortione ‘Phaeotrichoconis’ crotalariae. In vitro (MEA): Mycelium ex hyphis ramosis, septatis, laevibus, pallide brunneis, 1–2 μm latis compositum. Conidiophora ex hyphis oriunda, solitaria, erecta, cylindrica, recta vel leviter flexuosa, cellula basali bulbosa, sine rhizoideis, 4–6 μm lata, ad basim 10–15 μm lata, stipite modice brunneo vel atro-brunneo, laevi, transverse 2–5-euseptato, (15–)30–50(–60) μm longo, (3–)4(–5) μm lato, ad apicem cum (1–)2–4(–6) cellulis conidiogenis. Cellulae conidiogenae subcylindricae, allontoides vel doliiformes, rectae vel leviter curvatae, pallide brunneae, 5–8 × 2–2.5 μm, polyblasticae, sympodialiter proliferantes. Conidia hyalina, mucilagine aggregata (sed non catenata), ellipsoidea, apice subobtuso, basi subtruncata, latitudine maxima in parte centrali vel in parte supra centrum, saepe leviter asymmetrica, (3.5–)4(–4.5) × 2(–2.5) μm.

Etymology. Named after the host genus on which the fungus occurs, Acacia.

Leaf spots medium to dark brown, margin raised, red-purple, oblong to ellipsoid, up to 7 mm diam, associated with ‘Phaeotrichoconis’ crotalariae. Description based on culture on MEA: Mycelium consisting of branched, septate, smooth, pale brown, 1–2 μm wide hyphae. Conidiophores arising from mycelium, solitary, erect, cylindrical, straight to somewhat flexuous; basal cell bulbous, without rhizoids, 4–6 μm wide in upper part, but becoming 10–15 μm wide at basal part; stalk medium to dark brown, smooth, transversely 2–5-euseptate, (15–)30–50(–60) μm tall, (3–)4(–5) μm wide in the middle part; upper cell giving rise to (1–)2–4(–6) conidiogenous cells. Conidiogenous cells subcylindrical to allantoid or doliiform, straight to gently curved, pale brown, 5–8 × 2–2.5 μm; polyblastic, proliferating sympodially. Conidia hyaline, aggregating in slimy mass (never in chains), ellipsoid, apex subobtuse, base subtruncate, widest in the middle or upper third of the conidium, frequently somewhat asymmetrical, (3.5–)4(–4.5) × 2(–2.5) μm.

Characteristics in culture — Colonies on MEA erumpent, spreading, with moderate aerial mycelium; surface folded, margin lobate, smooth; surface olivaceous-grey, outer margin iron-grey; reverse iron-grey; colonies reaching up to 20 mm after 1 mo. Colonies fertile on SNA, OA and MEA.

Specimen examined. Indonesia, Pelalawan, living leaves of Acacia auriculiformis, Mar. 2008, leg. M.J. Wingfield, isol. P.W. Crous, holotype CBS H-20198, culture ex-type CPC 15078 = CBS 124104.

Notes —Castañeda & Kendrick (1990) established the genus Lauriomyces, characterised by dark brown conidiophores, and a series of branches, giving rise to chains of hyaline conidia via sympodial conidiogenesis. Brycekendrickomyces is morphologically similar to Lauriomyces, which in turn resembles Haplographium. The genus Haplographium is based on H. delicatum. Its confused history is discussed in detail by Zucconi & Pagano (1993). Haplographium delicatum was originally described by Berkeley & Broome as having conidia in chains (Mason 1933), which Saccardo (1886) also reported for the type species. Hughes (1958) noted that Stilbum catenatum was an older name for H. delicatum, which led Holubová-Jechová (1973) to place this species in Haplographium, while Castañeda & Kendrick (1990) placed it in Lauriomyces. If Haplographium and Lauriomyces are synonymous, the older name, Haplographium, would have preference. However, as shown here, ‘Lauriomyces’ catenatus is not congeneric with other species of Lauriomyces, such as L. heliocephalus (Rao & de Hoog 1986, Castañeda & Kendrick 1990) and L. bellulus (Crous & Wingfield 1994), suggesting that the two genera are distinct, and that the name Haplographium catenatum should be resurrected. Data from this study, furthermore, suggest that the strains of H. catenatum included here, probably represent a species complex.

Brycekendrickomyces differs from Haplographium and Lauriomyces by the absence of an intricate conidiophore branching system, and in having conidia produced in slimy heads rather than in chains. Furthermore, it is not phylogenetically related to species of Lauriomyces or Haplographium presently known from culture (Fig. 1). Brycekendrickomyces is somewhat similar to Argopericonia (Sutton & Pascoe 1987), although the latter fungus produces hyaline, apical conidiogenous heads, and it has ellipsoidal, single to short catenate conidia, each with a prominent, globose guttule.

E.G. Simmons, Alternaria. An identification manual: 668. 2007

Type species. Chalastospora cetera (E.G. Simmons) E.G. Simmons.

Conidiophores solitary, brown, smooth, arising from surface hyphae or as short, lateral branches from ropes of aerial hyphae; short, subcylindrical to flask-shaped, 0–2-transversely euseptate, seldom once geniculate or branched. Conidiogenous cells integrated, terminal or conidiophores reduced to conidiogenous cells, monotretic, determinate to polytretic, sympodial, conidiogenous loci visible as minute pores, without or with somewhat darkened and slightly thickened rim. Conidia in acropetal, branched chains, narrowly ellipsoid to narrowly ovoid, pale to medium brown, rarely 1–3 transversely euseptate, generally lacking longitudinal or oblique septa; conidial apex functioning as secondary conidiophore, proliferating laterally.

(Jacz.) U. Braun & Crous, comb. nov. — MycoBank MB509518; Fig. 4

Fig. 4

Chalastospora gossypii (CBS 114810). a–f. Superficial mycelium on SNA showing conidiophores with branched conidial chains. — Scale bars = 10 μm.

Basionym. Cladosporium gossypii Jacz., Holopkovoe Delo 1929, 5–6: 564. 1929 and Trudy Byuro Priklad. Bot. 24 (5): 181–182. 1931.

= Cladosporium malorum Rühle, Phytopathology 21: 1146. 1931.

≡ Alternaria malorum (Rühle) U. Braun, Crous & Dugan, Mycol. Progr. 2: 5. 2003.

= Phaeoramularia kellermaniana Marasas & I.H. Bredell, Bothalia 11: 217. 1974.

= Cladophialophora kellermaniana (Marasas & I.H. Bredell) U. Braun & Feiler, Microbiol. Res. 150: 83, 1995.

≡ Pseudocladosporium kellermanianum (Marasas & I.H. Bredell) U. Braun, A monograph of Cercosporella, Ramularia and allied genera 2: 393. 1998.

= Cladosporium porophorum Matsush., Icones Microfungorum a Matsushima lectorum: 36. 1975.

Characteristics in culture — See Braun et al. (2003).

Specimens examined. Canada, Saskatchewan, Matador, from grass litter, 27 May 1968, G.C. Bhatt 255, IMI 144487 = ATCC 38025 = CBS 597.69; from (?) soil, 18 Sept. 1973, H.A.H. Wallace, IMI 179345; Alberta, from Bromus inermis, 1994, R.J. Howad 397, IMI 360655, HAL. – Central Asia (without detailed locality), on fibres of Gossypium sp., 1927 and 1928, V.S. Fedorov, LEP, syntypes of Cladosporium gossypii. – Lebanon, from soil, July 1987, F. Seigle-Murandi, ATCC 200938 = CBS 900.87. – Libya, from Prunus persica, April 1975, Casay, IMI 194863. – New Zealand, Wellington, 40 Epuni Street, Te Aro Valley, wood and wallpaper from inside walls of a dwelling, 5 Sept. 2008, leg. D. De Vanny, isol. C.F. Hill 2008/3899, CPC 15567; Auckland, Henderson Valley Road, Henderson, leaves of Anethum graveolens (dill) along with Itersonilia perplexans, 1 Dec. 2003, leg. J. Pike, isol. C.F. Hill, MAF 943 = CBS 114809; Auckland, 90 Aberdeen Road, Castor Bay, isolated from Quercus robur (oak) leaves in association with Tubakia dryina, 5 Sept. 2008, leg. H. Nettleton, isol. C.F. Hill, MAF 954 = CPC 15567 = CBS 114810. – Pakistan, Karachi, from stored grains, 5 Jan. 1967, S.S. Hussain, IMI 124270. – South Africa, Western Cape Province, Kopgat, Calvinia, from wheat stubble, Feb. 1972, W.F.O. Marasas OP-76, PREM 44703, IMI 165252, cultures ATCC 28332 = IMI 165252 = PRE 44703 = CPC 3680 = CBS 266.75, ex-type cultures of Phaeoramularia kellermaniana. – Syria, from agricultural soil, Feb. 1980, M.I.A. Abdel-Kader, CPC 3685 = ATCC 200939 = CBS 173.80. – Turkey, Manisa, from Hordeum sp., 16 June 1971, Maksu & Selçuc, IMI 159198; Gossypium seeds, M. Esentepe, CBS 540.75. – USA, New Mexico, Red River, from a polypore on Picea sp., 4 Sept. 1996, D. Wicklow, IMI 386094; Washington State, from Bing cherry fruit, June 1992, F.M. Dugan, ATCC 96020; from fruits of Malus domestica, F.D. Heald, ATCC 36953; Washington State, Festuca idahoensis, F.M. Dugan, STE-U 4572 = CBS 114005; Pacific Northwest, Feb. 1966, C.W. Hesseltine, NRRL W 52-29 = CPC 3690 = CBS 148.66; Oregon, Portland, Triticum aestivum grain, June 1965, C.W. Hesseltine, NRRL A-13702 = CBS 216.65; Malus sylvestris fruit, Jan. 1931, F.D. Head, ATCC 36953 = MUCL 10096 = CBS 135.31; Washington State, Bromus tectorum, F.M. Dugan, CPC 4571 = CBS 112844; Washington State, Roza Canal near Prosser, isolated from dormant buds (overwintered) of Vitis vinifera, Mar. 2001, F.M. Dugan, holotype WSP 70286, cultures ex-type STE-U 4570 = CBS 112048 (var. polymorpha). – Uzbekistan, Bukhara, Experiment Station, on fibres of Gossypium hirsutum, 1928, V.S. Zelenetzi, LEP, lectotype of Cladosporium gossypii (selected here) (isolectotype in LEP); Bukhara, Shafrikanskoje, on fibres of Gossypium hirsutum, 1928, V.S. Zelenetzi, LEP, syntype of Cladosporium gossypii.

Notes — The genus Chalastospora appears to represent an anamorph lineage in the Pleosporales (Fig. 1). Chalastospora cetera and C. gossypii are clearly congeneric (Fig. 2). Based on the ITS data, there are some point mutations among strains of C. gossypii, suggesting that other genes need to be sequenced to fully elucidate the variation within this species (Fig. 2). On SNA, ramoconidia of CBS 114810 were 10–17 × 3–5 μm, and conidia narrowly ellipsoid-ovoid, cylindrical to fusiform, 6–10 × 2–2.5 μm, thus much smaller than that reported by Braun et al. (2003) on PDA. Jaczewski introduced the name Cladosporium gossypii in 1929, and provided a brief Russian description, including shape and size of conidia. This description, published before 1935, is, however, valid. In his paper of 1931, he re-introduced C. gossypii together with a Latin description and a micrograph of conidia. Type material of C. gossypii was re-examined and it is identical to C. malorum. However, C. gossypii is an older name than C. malorum, which was published in 1931, and has priority.

Crous & U. Braun, sp. nov. — MycoBank MB509519; Fig. 5

Fig. 5

Chalastospora ellipsoidea (CBS 121331). a–f. Superficial mycelium on SNA showing conidiophores with conidial chains; g, h. microconidiophores; i, j. conidia in chains. — Scale bars = 10 μm.

Chalastosporae gossypii similis, sed conidiis ellipsoideis, longioribus et leviter latioribus, (8–)10–15(–17) × 3(–3.5) μm.

Etymology. Named after its ellipsoid conidia.

On SNA: Conidiophores arising singly from aerial and creeping hyphae; subcylindrical, erect, medium brown, smooth, up to 25 × 3 μm, frequently reduced to conidiogenous cells, 5–13 × 3 μm; seldom once geniculate, mostly straight, with a slight swelling in the apical conidiogenous region; conidiogenous loci 1–3 per conidiogenous cell, medium brown, slightly thickened, darkened, up to 1 μm wide. Ramoconidia (0–)1–3-septate, ellipsoid-ovoid, subcylindrical or fusiform, smooth, medium brown, (12–)15–18(–30) × 3(–4) μm; apex at times with short beak, giving rise to lateral branch. Conidia ellipsoid to fusoid, medium brown, smooth, in long acropetal chains, simple, or branched with short apical or basal, lateral branches, (8–)10–15(–17) × 3(–3.5) μm, 0–1(–2)-septate; hila thickened and darkened, 0.5–1 μm wide.

Characteristics in culture — Colonies on OA spreading, with moderate, flattened aerial mycelium, smoke-grey. On MEA cinnamon with patches of hazel on surface and reverse. On PDA olivaceous-grey, with moderate aerial mycelium; iron-grey in reverse.

Specimen examined. Australia, on Triticum, H.L. Harvey & S. Perth, holotype CBS H-20199, culture ex-type E.G.S. 22.060 = CBS 121331.

Notes — The most characteristic features of this species are its short lateral branches, and ellipsoid conidia. It is clearly distinct from C. cetera and C. gossypii based on ITS sequence data (Fig. 2).

Crous & U. Braun, sp. nov. — MycoBank MB509520; Fig. 6

Fig. 6

Chalastospora obclavata (CBS 124120). a, b. Superficial mycelium on SNA showing conidiophores with branched conidial chains; c–e. conidia in chains. — Scale bar = 10 μm.

Differt ab omnibus specibus Chalastosporae conidiis intercalaribus obclavatis.

Etymology. Named after its obclavate conidia.

Sporulating poorly on SNA. Conidiophores 17–30 × 3–4 μm, arising singly from aerial and creeping hyphae; subcylindrical, somewhat clavate near apex of conidiogenous region, erect, straight to once geniculate, medium brown, smooth, frequently reduced to conidiogenous cells, 5–10 × 3–4 μm; conidiogenous loci medium brown, slightly thickened, darkened, 1–1.5 μm wide. Ramoconidia medium brown, smooth, developing short lateral beaks at apex that give rise to lateral chains (verticillate-like appearance), obclavate, widest at base, 0–3-septate, (28–)30–35 × (3.5–)4–5(–6) μm. Conidia obclavate, widest at base, (23–)26–30(–35) × (3.5–)4 μm, 0–3-septate; hila thickened, darkened, 1–1.5 μm wide.

Characteristics in culture — Colonies on OA spreading, with moderate, white aerial mycelium, grey-olivaceous to smoke grey; reverse grey-olivaceous. On MEA cream with dense aerial mycelial mat.

Specimen examined. USA, Kansas, Manhattan, ex air, Jan. 1958, C.T. Rogerson, holotype CBS H-20200, culture ex-type E.G.S. 12.128 = CBS 124120.

Notes — The most characteristic features of this species are its conidial branching pattern and conidial shape. This strain was discussed by Simmons under Alternaria cetera (Simmons 1996), and under Chalastospora in Simmons (2007). It is clearly distinct from C. cetera (ex-type CBS 121340, Fig. 7), C. ellipsoidea and C. gossypii based on ITS sequence data (Table 1, Fig. 2).

Fig. 7

Chalastospora cetera (CBS 121340). a–g. Superficial mycelium on SNA showing conidiophores with conidial chains. — Scale bars = 10 μm.

Intercalary conidia usually longer than 20 μm ……………2

Intercalary conidia shorter than 20 μm ……………3

Intercalary conidia narrowly ellipsoid to narrowly ovoid, widest in middle or lower third, (10–)19–24(–30) × 3(–4) μm, 0–3-septate ……………C. cetera

Intercalary conidia obclavate, widest at base, (23–)26–30 (–35) × (3.5–)4 μm, 0–3-septate ……………C. obclavata

Intercalary conidia narrowly ellipsoid-ovoid to cylindrical or fusiform, 6–10 × 2–2.5 μm, mostly aseptate ……C. gossypii

Intercalary conidia ellipsoid, not cylindrical nor fusiform, (8–)10–15(–17) × 3(–3.5) μm, 0(–2)-septate ……C. ellipsoidea

G.A. de Vries, Mycopathol. Mycol. Appl. 16: 47. 1962

Type species. Cyphellophora laciniata G.A. de Vries.

Hyphae fertile, pale brown, 1.5–3 μm wide, at times constricted at septa. Conidiogenous cells phialidic, intercalary, at times on short lateral branches, with a prominent to indistinct collarette. Conidia sickle-shaped, brown, smooth-walled, 1–3-septate, adhering in bundles.

Crous & Alfenas, sp. nov. — MycoBank MB509521; Fig. 8

Fig. 8

Cyphellophora eugeniae (CBS 124105). a, b. Colonies sporulating on OA; c–e. conidia attached to conidiogenous cells (arrows denote loci); f–j. conidia. — Scale bars = 10 μm.

Cyphellophorae taiwanensis similis, sed conidiis valde longioribus, (40–)60– 75(–90) × 2–2.5(–3) μm.

Etymology. Named after the host on which it occurs, Eugenia.

On PDA. Mycelium consisting of branched, greenish brown, septate, smooth, 3–5 μm wide hyphae, constricted at septa. Conidiogenous cells phialidic, intercalary, inconspicuous to subdenticulate, 1 μm wide, with minute collarettes, with several loci aggregated at hyphal swellings. Conidia subcylindrical, tapering towards obtuse ends, curved, smooth, hyaline to olivaceous, finely guttulate, 4–6(–10)-septate, prominently constricted at septa, widest in the middle of conidium, (40–)60–75(–90) × 2–2.5(–3) μm; conidia also anastomose and undergo microcyclic conidiation in culture.

Characteristics in culture — Colonies on PDA erumpent, with sparse aerial mycelium and even margins; surface olivaceous-grey, with patches of iron-grey; reverse iron-grey. On MEA erumpent, with folded surface and smooth, lobate margin, and sparse aerial mycelium; surface pale olivaceous-grey to olivaceous-grey; reverse iron-grey. On OA spreading, flat, with even, smooth margins and sparse aerial mycelium, olivaceous-grey. Colonies reaching 15 mm diam after 1 mo at 25 °C, fertile, sporulating in slimy sporodochial masses.

Specimen examined. Brazil, Rio Grande do Sul, Guaiba, living leaves of Stenocalyx uniflorus, 1 Apr. 2008, leg. A.C. Alfenas, isol. P.W. Crous, holotype CBS H-20201, culture ex-type CPC 15172 = CBS 124105.

Notes — The indistinct conidiogenous loci of C. eugeniae are reminiscent of those of C. taiwanensis (Matsushima 1985). The two species can be distinguished by the much longer conidia in C. eugeniae. Based on the key provided by Decock et al. (2003), C. eugeniae appears to represent a new species. Further collections of this complex are required to confirm the synonymy of the genera Cyphellophora with Pseudomicrodochium and Kumbhayama (Decock et al. 2003, Crous et al. 2007b), which were originally distinguished based on the absence of conidial pigmentation. The ITS sequence of C. eugeniae has 89 % similarity to that of Cyphellophora hylomeconis (GenBank accession EU035415).

Phialides intercalary, reduced to a sessile locus with collarette …………2

Phialides prominent, cylindrical, flask-shaped, sessile or with an elongated base …………6

Conidia 1–3-septate…………3

Conidia usually more than 3-septate…………4

Conidia up to 2.5 μm wide (11–20 × 2–2.5 μm), 1(–2)-septate…………C. fusarioides

Conidia up to 5 μm wide (11–25 × 2–5 μm), 1–3-septate………… C. laciniata

Conidia up to 2 μm wide, 3–6-septate, sigmoid (16–35 × 1.5–2 μm)………… C. taiwanensis

Conidia wider than 2 μm …………5

Conidia subcylindrical, 4–6(–10)-septate, (40–)60–75(–90) × 2–2.5(–3) μm ………… C. eugeniae

Conidia sigmoid, 1–5-septate, (15–)25–35(–55) × (2.5–) 3(–4) μm ………… C. hylomeconis

Phialides short to long and cylindrical; conidia 1–1.2 μm wide, 2–3-septate …………C. suttonii

Phialides prominent, flask-shaped, sessile or with an elongated base …………7

Conidia mainly straight, on average smaller than 20 μm, 1–5-septate …………C. pluriseptata

Conidia straight to more commonly falcate, curved, or sigmoid, on average longer than 20 μm…………8

Conidia (1–)3-septate, wider than 3 μm, 25–40 × 3.5–5.5 μm; phialides commonly with an elongated base …… C. indica

Conidia 2–8-septate, narrower than 3 μm; phialides without elongated base …………9

Conidia vermiform, mostly curved, mostly 4–8-septate, 30– 55 × 1.2–1.5 μm ………… C. vermispora

Conidia straight, falcate or slightly sigmoid, (2–)3–6-septate, (18–)19.5–28(–29) × 1.5–2 μm ………… C. guyanensis

Corda, in Weitenweber, Beitr. Gesammten Natur-Heilwiss., Prag 1: 87. 1836

Type species. Dictyosporium elegans Corda.

Conidiomata sporodochial, black, scattered. Mycelium predominantly immersed, consisting of branched, septate, smooth, thin-walled hyphae. Conidiophores micronematous, mononematous, pale brown, smooth to finely verruculose, thin-walled, septate, cylindrical. Conidiogenous cells monoblastic, integrated, pale to medium brown, smooth to finely verruculose, cylindrical, determinate; at times remaining attached to released conidium. Conidia cheiroid, medium to dark brown, smooth, euseptate, one cell-layer thick, cells arranged in 1–2 planes, fan-shaped; cell rows originating from a central basal cell; rows usually attached along their length; outer rows usually shorter than inner rows, at times paler in colour than central rows, and with or without hyaline, thin-walled, 1–2-celled appendages that are allantoid, clavate to globose, or fusoid to cylindrical.

Crous & A.R. Wood, sp. nov. — MycoBank MB509522; Fig. 9

Fig. 9

Dictyosporium strelitziae (CBS 123359). a. Colony sporulating on PDA; b, c. conidia attached to conidiogenous cells; d–h. conidia with hyaline, apical appendages. — Scale bars = 10 μm.

Dictyosporii bulbosi valde simile, sed conidiis leviter longioribus, (30–)40– 46(–55), et phylogenetice manifeste divergens.

Etymology. Named after the host genus Strelitzia, on which it occurs.

Leaf spots absent, colonies occurring on dead leaf tissue. Description based on colonies sporulating on WA with pine needles (colonies also sporulate well on OA and MEA): Mycelium predominantly internal in host tissue, consisting of branched, septate, smooth, brown, 2–2.5 μm wide hyphae. Conidiomata sporodochial, scattered, black, up to 170 μm diam. Conidiophores subcylindrical, darker brown than hyphae, at times slightly verruculose, irregularly curved to geniculate-sinuous, 1–3-septate, 10–25 × 2–2.5 μm; older conidiophores curved like sheperd’s crook. Conidiogenous cells terminal, medium brown, verruculose, subcylindrical, curved (semi-circular), 5–10 × 2–2.5 μm. Conidia solitary, complanante, cheiroid, smooth-walled, uniformly pale brown, becoming uniformly medium brown at maturity; cells arranged in (4–)5(–6) rows, meeting at basal cell; outer rows with 8–10 cells, with a hyaline, globose, apical appendage, 5–10 μm diam; outer rows shorter than inner rows; inner rows with 7–11 cells; central row with 6–10 cells; conidia (30–)40–46(–55) × (20–)21–23(–25) μm.

Characteristics in culture — Colonies on OA flat, spreading, without aerial mycelium, and with regular, even margin; on MEA flat, spreading, with moderate aerial mycelium and regular, smooth margin; surface buff, reverse cinnamon; colonies on both media reaching 30 mm diam after 1 mo at 25 °C.

Specimen examined. South Africa, KwaZulu-Natal, Skyline Nature Reserve, Uvongo, on dead leaves of Strelitzia nicolai, 29 May 2008, leg. A. Wood, isol. P.W. Crous, CBS H-20202 holotype, cultures ex-type CPC 15359–15361, CBS 123359.

Notes — The genus Dictyosporium is well defined, and separated from similar genera by having smooth-walled, euseptate conidia produced from determinate conidiogenous cells (Sutton et al. 1996, Tsui et al. 2006). Based on the key provided by Cai et al. (2003b), D. strelitziae is morphologically most similar to D. bulbosum (conidia 27–46 × 11–30 μm), but its conidia are somewhat longer, and there is a 10 bp difference between the ITS sequences of D. strelitziae and D. bulbosum (DQ018086). Phylogenetically, D. strelitziae is closest to D. elegans (conidia 44–80 × 24–36 μm; appendages absent) (5 bp difference in the ITS sequence, DQ018087), but it has smaller conidia than the latter species. Furthermore, it also appears distinct from all species not occurring in the key of Cai et al. (2003b) (Arambarri et al. 2001, Cai et al. 2003a, Zhao & Zhang 2003, Kodsueb et al. 2006, Cai & Hyde 2007, McKenzie 2008).

Conidia with appendages………… 2

Conidia lacking appendages………… 13

Appendages apical………… 3

Appendages not apical………… 4

Apical appendages aseptate………… 6

Apical appendages frequently 1-septate, cylindrical, 24–51 × 6–10.5 μm; conidia 27.5–47.5 × 20–25 μm, complanate, with 4–5 rows of cells …………D. canisporum

Appendages subapical, cylindrical to clavate; conidia 52.5– 72.5 × 18.5–26.5 μm, not complanate, with 5 rows of cells …………D. tetraploides

Appendages not subapical, but central or basal………… 5

Appendages central, hyaline, thin-walled, clavate to obovoid; conidia 36–45 × 16–21 μm, not complanate, mostly 7 rows of cells …………D. musae

Appendages basal, fusoid to cylindrical; conidia 22–28 × 12.5– 18 μm, complanate, with 3 rows of cells …D. manglietiae

Conidia with 3 rows of cells, (27–)31–43 × 10–12 μm ………D. freycinetiae

Conidia with more than 3 rows of cells …………7

Conidia mostly with 4 rows of cells …………8

Conidia with 5 or more rows of cells………… 10

Conidia with darker colour at apex of inner rows; apical cells of outer rows each bearing a hyaline, cylindrical appendage………… D. nigroapice

Conidia concolorous………… 9

Conidia 24–40 × 14–20 μm; appendages clavate …………D. tetraseriale

Conidia 36–45 × 16–21 μm; appendages tapering………… D. palmae

Conidia mostly comprising 5 rows of cells …………11

Conidia mostly comprising 6–8 rows, 46–88 × 26–46 μm; appendages hyaline, curved………… D. digitatum

Conidia longer than 32 μm, appendages globose to obovoid …………12

Conidia shorter than above, 26–32 × 15–24 μm; appendages cylindrical to clavate………… D. alatum

Conidia up to 46 μm long, and 30 μm wide, 27–46 × 11–30 μm; appendages globose to obovoid ……D. bulbosum

Conidia longer than 46 μm, but not wider than 25 μm, (30–)40–46(–55) × (20–)21–23(–25) μm; appendages globose………… D. strelitziae

Conidia complanate, one cell layer thick………… 14

Conidia not complanate, more than one cell layer thick ……24

Conidia regularly consisting of 3 rows of cells …………15

Conidia consisting of at least 4 rows of cells………… 16

Conidia 15–22.5 × 10–16.5 μm ……D. lakefuxianensis

Conidia 26–32 × 16–18 μm………… D. triseriale

Conidia curved, with 5–7 rows of cells, each curving in the same direction, 34–56 × 20–38 μm ……D. foliicola

Conidia not curved………… 17

Conidia less than 25 μm long………… 18

Conidia more than 25 μm long …………19

Conidia 18–24 × 13–19 μm ……D. brahmaswaroopii

Conidia 15–17 × 11–12 μm ……D. schizostachyfolium

Conidia with paler outer rows …………20

Conidia concolorous …………21

Conidia 25–45 × 22–38 μm, with (5–)6(–7) rows …………D. yunnanensis 1

Conidia 26–40 × 13–25 μm, mostly with 5 rows …………D. zeylanicum

Conidia with 4 rows, 23.5–40 × 16–21.5 μm …………D. tetrasporum

Conidia with more than 4 rows …………22

Conidia 40–80 × 24–36 μm, mostly with 5 rows, slightly constricted at septa………… D. elegans

Conidia mostly with more than 5 rows, strongly constricted at septa………… 23

Conidia 26–34 × 23–34 μm, mostly with 7–9 rows of cells; conidiomata sporodochial …………D. polystichum

Conidia 38–56 × 25–32 μm, mostly 6–8 rows of cells; conidiomata not sporodochial…………D. toruloides

Conidia campaniform, with a darker base; with 12–16 rows of cells, 22–40 × 20–30 μm………… D. campaniforme

Conidia more or less cylindrical, concolorous, comprising 3–7 rows of cells …………25

Conidia regularly with 3 rows of cells; usually 13.5 μm or less wide………… 26

Conidia mostly with 4–7 rows of cells; more than 13.5 μm wide………… 28

Conidia 40–60 × 10–13.5 μm…… D. triramosum

Conidia shorter than 43 μm ………27

Conidia 36–43 × 11–12 μm; sporodochia usually covered with gelatinous matrix……… D. australiense

Conidia 20–30 × 10–12 μm; sporodochia not as above ……D. micronesicum

Conidia 40–50 × 18–25 μm, with 4–6 rows of cells, muriform, with hyaline, subglobose conidiogenous cell remaining attached as basal appendage ………D. gauntii

Conidial morphology not as above ………29

Conidia with rows of cells that are distinctly incurved or hook-like at the apex……… 30

Conidia with more or less straight rows of cells at the apex ………32

Conidia 105–121 × 25–32 μm ………D. giganticum

Conidia up to 80 μm long ………31

Conidia 50–80 × 20–30 μm……… D. heptasporum

Conidia 33–42 × 16–20 μm……… D. subramanianii

Colonies effuse, not sporodochial; conidia irregularly cylindrical or oblong, strongly constricted at septa; 30–50 × 12–30 μm ………D. oblongum

Colonies sporodochial; conidia more or less cylindrical, slightly constricted at septa, 53–76 × 19–22 μm……… D. cocophilum

M.C. González, Anaya, Glenn, Saucedo & Hanlin, Mycotaxon 101: 254. 2007.

Type species. Edenia gomezpompae M.C. González, Anaya, Glenn, Saucedo & Hanlin.

Conidiophores fasciculate, subcylindrical, medium brown, finely roughened, 3–15-septate, straight to variously curved or geniculate-sinuous, irregular in width, constricted at some septa, with percurrent rejuvenation in upper part, situated on a submerged, brown stroma. Conidiogenous cells terminal, integrated, becoming paler brown towards apex, tapering to a subtruncate tip, with several lateral loci that are somewhat thickened and protruding (pimple-like), giving rise to conidia via sympodial proliferation near apex. Conidia 11–16 × 3.5–6 μm, subhyaline, smooth, thin-walled, finely guttulate, fusoid-ellipsoidal with obtuse apex and tapering from its widest point in the middle towards a subtruncate base, 1–1.5 μm wide.

M.C. González, Anaya, Glenn, Saucedo & Hanlin, Mycotaxon 101: 254. 2007 — Fig. 10

Fig. 10

Edenia gomezpompae (CBS 124106). a. Hyphal tufts visible when cultivated on MEA; b. leaf spot with conidiophores; c. fasciculate conidiophores; d. conidiophores arising from conidioma; e–g. conidiophores and conidiogenous cells; h. conidia; i. conidiomata forming on OA; j. conidioma with ostiolar setae; k, l. conidiogenous cells; m. conidia. — Scale bars = 10 μm.

Leaf spots subcircular, 3–12 mm diam, grey-brown, with a dark brown, raised border, surrounded by a diffuse, black halo (absent in smaller spots). Conidiophores in fascicles of 5–30, subcylindrical, medium brown, finely roughened, 3–15-septate, straight to variously curved or geniculate-sinuous, 50–170 × 4–6 μm, irregular in width, constricted at some septa, with percurrent rejuvenation in upper part; fascicles randomly distributed over lesion, amphigenous, visible as erect, dark brown to black tufts on lesions, situated on a submerged, brown stroma, up to 60 μm wide and 40 μm high, intermingled among leaf trichomes (fruiting structures of a Ramularia sp. and ascomata of another fungus also present in some lesions). Conidiogenous cells 15–30 × 3–4 μm, terminal, integrated, becoming paler brown towards apex, tapering to a subtruncate tip, with several lateral loci that are somewhat thickened and protruding (pimple-like), up to 1 μm diam, giving rise to conidia via sympodial proliferation near apex, but some conidiogenous cells also show signs of percurrent proliferation, but this appears to be linked to rejuvenation, not conidiogenesis. Conidia (11–)13–15(–16) × (3.5–)4.5–5.5(–6) μm, subhyaline, smooth, thin-walled, finely guttulate, fusoid-ellipsoidal with obtuse apex and tapering from its widest point in the middle towards a subtruncate base, 1–1.5 μm wide.

Characteristics in culture — Colonies fluffy, with white hyphal strands that turn brown with age; surface woolly with abundant aerial mycelium; margins uneven. On MEA buff to rosy-buff (surface), brick to dark brick (reverse); on PDA fluffy, cream to buff (surface), dark brick to buff (reverse); on OA brick with patches of cream to buff. Colonies reaching 25 mm diam after 2 wk at 25 °C, becoming fertile on OA.

Specimens examined. Mexico, Quintana Roo, Isla Mujeres Municipality, El Eden Ecological reserve, from leaves of Callicarpa acuminata (Lamiaceae), May 2002, A. Saucedo-García & A.L. Anaya, holotype MEXU 25346. – Philippines, on Senna alata (≡ Cassia alata) (Caesalpiniaceae), Oct. 2008, leg. C.J.R. Cumagun, isol. P.W. Crous, epitype designated here CBS H-20203, cultures CPC 15689 = CBS 124106, CPC 15690, 15691.

Notes — The genus Edenia was originally introduced for a sterile fungus (suspected to be a member of the Pleosporaceae), isolated as an endophyte from leaves of Callicarpa acuminata in Mexico (González et al. 2007). The genus was characterised by producing numerous sterile, whitish mycelial strands and coils on PDA. The present collection from Cassia alata in the Philippines has the same colony characteristics, and based on its identical DNA sequence data (GenBank EF565744.1), we believe that this is the same fungus. What is interesting, however, is the fact that the latter collection was made from conidia of a dematiaceous hyphomycete sporulating on leaf spots of C. alata. As other fungi were also present on these spots, its potential role as pathogen remains uncertain. On host tissue, however, some conidiophores were associated with a weakly developed layer of pale brown stromatic cells. On OA, cultures became fertile, and conidiophores were arranged around well-developed ostioles of submerged pycnidia (with a similar pale brown stromatic wall to that observed on the host). It is possible, therefore, that if the field material had been placed in moist chambers, the pycnidial state would have developed. The latter state resembles species that are pyronellea-like in morphology.

Morphologically, the hyphomycete state of Edenia resembles genera such as Digitopodium, although species of this genus have rhizoids, and 1-septate, pale brown conidia that can also occur in short chains (Heuchert et al. 2005). It also shares some similarities with Blastophorum (Matsushima 1971), although the latter fungus is distinct in having solitary conidiophores with rhizoids, and a hyaline, upper conidiogenous region.

B. Sutton, Trans. Brit. Mycol. Soc. 61: 426. 1973

Type species. Thedgonia ligustrina (Boerema) B. Sutton.

Conidiomata fasciculate, punctiform. Mycelium internal, hyphae subhyaline, septate, branched, forming substomatal stromata, hyaline to pale brown. Conidiophores fasciculate, arising from stromata, simple, rarely branched, subcylindrical, straight to geniculate-sinuous, continuous to septate, smooth, hyaline to pale yellowish green. Conidiogenous cells integrated, terminal, occasionally conidiophores reduced to conidiogenous cells, sympodial, conidiogenous loci more or less planate, unthickened, non-pigmented. Conidia in disarticulating chains, rarely in branched chains, subcylindrical to obclavate, with one to several transverse eusepta, hyaline or almost so, apex rounded to truncate, base truncate, hila flat, unthickened, hyaline.

(Boerema) B. Sutton, Trans. Brit. Mycol. Soc. 61: 428. 1973 — Fig. 11

Fig. 11

Thedgonia ligustrina (CBS 124332). a, b. Leaf spots on Ligustrum; c. fasciculate conidiophores; d, e. conidiophores; f, g. conidia. — Scale bars = 10 μm.

Basionym. Cercospora ligustrina Boerema, Tijdschr. Plantenziekten 68: 117. 1962.

≡ Cercoseptoria ligustrina (Boerema) Arx, Genera of Fungi Sporulating in Pure Culture, ed. 3: 306, Lehre 1981.

Characteristics in culture — On MEA erumpent, slow growing, 5–8 mm after 2 wk, with moderate, white aerial mycelium and smooth, lobate margins; umber in reverse. On OA 5–8 mm diam after 2 wk, submerged to flattened on surface, sparse aerial mycelium, and smooth, even margins; umber on surface.

Specimens examined. Asia, on Ligustrum sp., H. Evans, CPC 4296 = W2072, CPC 4297 = W 2073, CPC 4298 = W 1877. – Netherlands, Eefde, on Ligustrum ovalifolium, 23 Mar. 1959, G.H. Boerema, holotype L, ex-type culture CBS 148.59; Bilthoven, on L. ovalifolium, 2003, P.W. Crous, CPC 10530 = CBS 124332, CPC 10532, 10533. – South Korea, Namyangju, on L. ovalifolium, 9 Oct. 2002, leg. H.D. Shin, isol. P.W. Crous, CBS H-20204, CPC 10019, 10861–10863; Suwon, on L. obtusifolium, 2 Oct. 2007, leg. H.D. Shin, isol. P.W. Crous, CBS H-20207, CPC 14754–14756.

Notes — Kaiser & Crous (1998) linked ‘Thedgonia’ lupini as anamorph to Mycosphaerella lupini, and thus suggested that Thedgonia belongs in the Mycosphaerellaceae. Results of this study (Fig. 1), however, show that Thedgonia s.str. belongs to the Helotiales, and is unrelated to the Mycosphaerellaceae. Furthermore, there is presently no separate anamorph genus in the Mycosphaerellaceae to accommodate ‘T.’ lupini. Although ‘T.’ lupini resembles species of Pseudocercosporella (Braun 1995), it appears to represent a separate phylogenetic lineage.

R.T. Moore, Mycologia 47: 90. 1955

Type species. Trochophora fasciculata (Berk. & M.A. Curtis) Goos (syn. T. simplex (Petch) R.T. Moore).

Colonies hypophyllous, medium to dark brown, consisting of numerous synnemata. Stroma absent, but a superficial network of hyphae linking the various synnemata. Conidiophores synnematous, mostly unbranched and straight, or with 1–2 short branches, straight or curved, cylindrical, individual conidiophores tightly aggregated, but separating near the apex, pale to medium brown, smooth. Conidiogenous cells polyblastic, integrated, terminal, determinate to sympodial, with visible unthickened scar, clavate. Conidia solitary, terminal or lateral on conidiogenous cells, prominently curved to helicoid, pale to medium brown, smooth, transversely septate with a darkened, thickened band at the septa.

(Berk. & M.A. Curtis) Goos (as ‘fasciculatum’), Mycologia 78: 759. 1986 — Fig. 12

Fig. 12

Trochophora fasciculata (CPC 10280). a. Leaf spots on Daphniphyllum; b. colony on MEA; c. fasciculate conidiophores; d. conidiophores and conidiogenous cells; e–g. conidia. — Scale bars = 10 μm.

Basionym. Helicoma fasciculatum Berk. & M.A. Curtis, U.S. North Pacific Exped.: 142. (1853–1856) 1853.

≡ Helicosporium fasciculatum (Berk. & M.A. Curtis) Sacc., Syll. fung. 4: 560. 1886.

≡ Helicomyces fasciculatus (Berk. & M.A. Curtis) Pound & Clem., Minnesota Bot. Stud. 1: 658. 1896.

= Helicosporium simplex Syd., Mém. Herb. Boissier 4: 7. 1900.

≡ Helicoma simplex (Syd.) Linder, Ann. Missouri Bot. Gard. 16: 315. 1929.

= Helicostilbe simplex Petch, Ann. Roy. Bot. Gard. Peradeniya 7: 321. 1922.

≡ Trochophora simplex (Petch) R.T. Moore, Mycologia 47: 90. 1955.

Specimen examined. Korea, Pusan, on leaves of Daphniphyllum macropodum, 13 Nov. 2002, leg. H.D. Shin, isol. P.W. Crous, KUS-F19414, cultures CPC 10280–10282.

Notes — Two species have been described in the genus, namely T. fasciculata and T. simplex; the latter recognised as a synonym of the former (Zhao et al. 2007). Within the Mycosphaerellaceae, pseudocercospora-like species cluster in two well-defined clades, namely the P. vitis clade (Pseudocercospora s.str.), and the P. heimii clade (pseudocercospora-like). Based on LSU DNA phylogeny (Fig. 1), Trochophora clusters basal to the pseudocercospora-like clade. Although it is tempting to use the name Trochophora for this clade, further collections of Trochophora are required to clarify the morphological variation among taxa with this unique conidial morphology. Using sequence data of the ITS gene, the closest taxa obtained from a BLAST search is the Mycosphaerella heimii species complex (96 % similarity).

Zhao et al. (2007) consider T. fasciculata as a pathogen of Daphniphyllum, and report it from this host in several Asian countries, namely Sri Lanka, China (incl. Hong Kong and Taiwan) and India.

D.E. Shaw & Alcorn, Austral. Syst. Bot. 6: 273. 1993

≡ Verrucispora D.E. Shaw & Alcorn, Proc. Linn. Soc. New South Wales 92: 171. 1967. (nom. illegit.).

Type species. Verrucisporota proteacearum (D.E. Shaw & Alcorn) D.E. Shaw & Alcorn.

Mycelium consisting of pale brown, septate, verrucose hyphae. Stroma forming in substomatal cavities, cells brown-walled, pseudoparenchymatous. Conidiophores macronematous, mononematous, simple, flexuous, often geniculate, septate, mainly smooth, pale to dark brown, tapering towards the apex, but often becoming more swollen, and also verruculose to verrucose at the apex. Conidiogenous cells cylindrical, becoming geniculate, integrated, terminal, becoming intercalary, polyblastic, proliferating sympodially, cicatrised; conidiogenous loci planate, conspicuous, protuberant, thickened and darkened. Conidia cylindrical, narrowing slightly to an obtuse apex and with a truncate base with a distinctly thickened hilum, medium brown, straight or curved, transversely septate, verrucose to verruculose.

(Cooke & Massee) Beilharz & Pascoe, Mycotaxon 82: 360. 2002

Basionym. Cercospora daviesiae Cooke & Massee, Grevillea 18: 7. 1889.

Teleomorph. Mycosphaerella daviesiicola Beilharz & Pascoe, Mycotaxon 82: 364. 2002.

Characteristics in culture — On MEA erumpent, spreading with folded surface, and sparse aerial mycelium and even, lobate margin; surface iron-grey to olivaceous-grey; reverse iron-grey; colonies reaching 7 mm diam after 2 wk. On PDA erumpent, spreading, with moderate aerial mycelium and uneven margins; surface white in middle, olivaceous-grey in outer region, iron-grey underneath; colonies reaching 8 mm diam after 2 wk. On OA erumpent, spreading, with moderate aerial mycelium and uneven margin; surface white in middle, olivaceous-grey in outer region; colonies reaching 8 mm diam after 2 wk.

Specimen examined. Australia, Victoria, on living leaves of Daviesia mimosoides (≡ D. cormybosa var. mimosoides), V. & R. Beilharz, VPRI 31767 = CBS 116002.

Notes — The type species of the genus Stenella, S. araguata, clusters in the Teratosphaeriaceae (Crous et al. 2007a), and thus the majority of the stenella-like anamorphs in the Mycosphaerellaceae, will need to be placed in another genus. One option would be Zasmidium (Arzanlou et al. 2007), which clusters in the Mycosphaerellaceae, along with Verrucisporota (Fig. 1). This clade, however, is neither morphologically nor phylogenetically well resolved, and taxa need to be added to improve the phylogeny before a reasonable assessment can be made. The ITS sequence of this species is distinct from the other two species of this genus treated in this paper (Table 1).

Crous & Summerell, sp. nov. — MycoBank MB509523; Fig. 13

Fig. 13

Verrucisporota grevilleae (CBS 124107). a. Leaf spots on Grevillea; b. conidiophores; c, d. conidiophores and conidiogenous cells; e–h. conidia; i. colony on PDA; j. colony on SNA. — Scale bars = 10 μm.

Differt a Verrucisporota protearum conidiis angustioribus et longioribus, (30–) 50–65(–80) × (5–)6–7 μm, et conidiophoris brevioribus, (35–)80–120(–160) × (5–)6–7 μm.

Etymology. Named after the host genus on which it occurs, Grevillea.

Leaf spots angular, elongated, amphigenous, 1–2 mm wide, 3–10 mm long, medium to dark brown to black, discrete. Mycelium immersed and superficial, hyphae medium brown, septate, verrucose, 1.5–3 μm wide. Stroma up to 60 μm wide and 40 μm high, forming in substomatal cavities, becoming erumpent, cells brown, thick-walled, pseudoparenchymatous. Conidiophores macronematous, mononematous, caespitose, emerging through the stomata, simple, flexuous, often geniculate-sinuous, 4–7-septate, mainly smooth, dark brown, from a bulbous base tapering towards the apex, but often becoming more swollen, and also verrucose at the apex, (35–)80–120 (–160) × (5–)6–7 μm. Conidiogenous cells cylindrical, becoming geniculate, integrated, terminal, polyblastic, proliferating sympodially, 20–45 × 5–7 μm, with conspicuous, cicatrised, protuberant, conidiogenous loci, 3 μm diam. Conidia subcylindrical, narrowing slightly to an obtuse apex (frequently swollen), and with a truncate base with a distinctly thickened, darkened, somewhat refractive hilum, 3 μm wide, red-brown, straight or curved, with 3–7(–12) mainly unconstricted eusepta, thick-walled, verrucose, (30–)50–65(–80) × (5–)6–7 μm. Conidiophores frequently arising from brown, erumpent spermatogonia, up to 150 μm wide. Spermatia hyaline, smooth, bacilliform, 4–6 × 1–1.5 μm.

Characteristics in culture — Colonies on MEA erumpent, with sparse aerial mycelium; margins feathery, crenate; surface folded, with zones of salmon or smoke-grey mycelium; outer region and reverse olivaceous-grey; colonies reaching 10 mm diam after 1 mo.

Specimen examined. Australia, Northern Territory, Emerald Springs (13°37′13.3″ 131°36′40″), on leaves of Grevillea decurrens, 22 Sept. 2007, leg. B. Summerell, isol. P.W. Crous, CBS H-20205, cultures CPC 14761 = CBS 124107, CPC 14762, 14763.

Notes — Conidia of V. grevilleae are narrower and longer, and conidiophores shorter than those of V. protearum (conidia 23–51 × 5.6–10.5 μm, conidiophores up to 290 μm long, 4.5– 8.5 μm wide; Shaw & Alcorn 1967). South African specimens from the genus Protea have conidia that are (20–)31–36(–49) × (7–)8.5–9.5(–12) μm (Crous et al. 2004a). These findings suggest that the fungus treated as V. protearum on Proteaceae (Shaw & Alcorn 1967, 1993, Beilharz & Pascoe 2002, Crous et al. 2004a), probably represents a complex of several taxa.

(D.E. Shaw & Alcorn) D.E. Shaw & Alcorn, Austral. Syst. Bot. 6: 273. 1993

Basionym. Verrucispora proteacearum D.E. Shaw & Alcorn, Proc. Linn. Soc. New South Wales 92: 171. 1967.

Characteristics in culture — On MEA erumpent with sparse aerial mycelium; surface cream to pale olivaceous-grey, folded, with smooth, even margin; reverse brown-vinaceous; reaching 8 mm diam after 2 wk. On PDA erumpent with sparse aerial mycelium and smooth to feathery margin; surface cream to pale olivaceous-grey; reverse olivaceous-grey, reaching 8 mm diam after 2 wk. On OA erumpent, with moderate aerial mycelium and uneven margin, pale white in middle, pale olivaceous-grey in outer region; reaching 10 mm diam after 2 wk.

Specimen examined. Australia, Grevillea sp., V. Beilharz, VPRI31812 = CBS 116003.

Notes — Because V. proteacearum was originally described from Finschia (conidia 23–51 × 5.6–10.5 μm; Shaw & Alcorn 1967), there is a strong possibility that the strain listed here from Grevillea (conidia 30–45 × 10–12 μm on OA) may represent a different taxon to the one occurring on Finschia. Although apparently identical based on the LSU phylogeny (see Fig. 1), the ITS sequence of this isolate is different to that of V. grevilleae (95 % similarity and 4 % gaps).

Conidia wider than 4.5 μm……… 2

Conidia narrower than 4.5 μm……… 3

Conidia up to 56 μm long ………4

Conidia longer than 56 μm, 3–7(–12)-septate, (30–)50– 65(–80) × (5–)6–7 μm; on Grevillea ………V. grevilleae

Conidia mostly up to 30 μm long, (0–)2–3(–7)-septate, 13–30(–70) × 2.75–4 μm; on Capparis .V. kimberleyana

Conidia longer, mostly up to 77 μm long, 1–11-septate, (10–)27–77(–108) × 3–4.5 μm; on Struthanthus……… V. struthanthicola

Conidia up to 3-septate, obclavate, 1–3-septate, 32.5–55 × 7–10.5 μm; on Celastrus ………V. indica

Conidia more than 3 septa ………5

Conidia up to 32 μm long; (1–)3–4(–5)-septate, 20–32 × 6–10 μm; on Bridelia……… V. brideliae

Conidia frequently longer than above……… 6

Conidia 0–6-septate, 18–56 × 4.5–7 μm; on Daviesia (Beilharz & Pascoe 2002)……… V. daviesiae

Conidia 3–7-septate, 23–51 × 5.6–10.5 μm; on Finschia……… V. proteacearum

Bat., Publ. Inst. Micol. Univ. Fed. Pernambuco 283: 5. 1960

Type species. Vonarxia anacardii Bat. & J.L. Bezerra.

Mycelium immersed and superficial, composed of branched, septate, pale to medium brown, smooth to finely roughened hyphae. Conidiomata sporodochial; basal stroma composed of globose-ellipsoidal, brown, slightly roughened cells. Setae irregularly scattered throughout colony, simple, subulate with a bulbous base, straight to slightly curved, dark brown, smooth to slightly roughened, thick-walled, 5–16-euseptate, septa rather thick, but becoming thinner towards apex. Conidiogenous cells arise from upper cells of the stroma, tightly aggregated, doliiform to ellipsoid, pale brown to subhyaline or hyaline, smooth, giving rise to a cluster of conidia by means of sympodial proliferation, with successive conidia forming at a higher level. Conidia hyaline, smooth-walled, tetraradiate, basal cell subcylindrical to clavate to doliiform, 0–1-septate; upper three arms arise from the apical part of the basal cell, 3–10-septate, subcylindrical to cylindrical, apex subobtuse.

(Speg.) Aa, Persoonia 13: 128. 1986 — Fig. 14

Fig. 14

Vonarxia vagans (CBS 123533). a, b. Colony on PDA; c. colony with setae; d, e. setae with rounded apices and swollen bases, lacking rhizoids; f–i. conidiogenous cells giving rise to conidia; j–o. conidia. — Scale bars = 10 μm.

Basionym. Ypsilonia vagans Speg., Revista Mus. La Plata, Secc. Bot. 15: 35. 1908.

≡ Kazulia vagans (Speg.) Nag Raj, Canad. J. Bot. 55: 1621. 1977.

On PDA. Mycelium immersed and superficial, composed of branched, septate, pale to medium brown, smooth to finely roughened, 3–5 μm wide hyphae. Conidiomata sporodochial, flattened to erect and globose (especially on WA, not so on MEA or PDA, tending to be more flattened, and more hemispherical on OA), up to 300 μm diam; basal stroma up to 70 μm thick, composed of globose-ellipsoidal, brown, slightly roughened cells, 5–10 μm diam. Setae irregularly scattered throughout colony, simple, subulate with a bulbous base, straight to slightly curved, dark brown, smooth to slightly roughened, thick-walled (1–1.5 μm diam), (5–)10–12(–16)-septate, septa rather thick, but becoming thinner towards apex, basal cell 10–13 μm wide, with slight taper towards bluntly rounded, obtuse apex, (120–)150–200(–220) μm; width at basal septum (5–)6(–7) μm; width at apical septum, 2–3(–5) μm; apical two cells frequently pale brown; individual cells 10–25 μm long. Conidiogenous cells arise from upper cells of the stroma, tightly aggregated, doliiform to ellipsoid, pale brown to subhyaline or hyaline, smooth, 8–10 × 3–5 μm, giving rise to a cluster of conidia by means of sympodial proliferation, with successive conidia forming at a higher level. Conidia hyaline, smooth-walled, tetraradiate, basal cell subcylindrical to clavate to doliiform, 0–1-septate, 10–15 × (1.5–)2–3 μm (10–18 μm long on OA); upper three arms arise from the apical part of the basal cell, 3–5-septate (prominently constricted at septa on WA and MEA, up to 10-septate on these media), subcylindrical to cylindrical, apex subobtuse, arms 20–55 μm long (20–90 μm on OA), 1.5–2 μm wide (2–3 μm wide on OA).

Characteristics in culture — Colonies on OA spreading, with sparse aerial mycelium, and uneven, striate surface, with crenate margin; surface black, with patches of mouse-grey, reaching up to 25 mm diam after 1 mo; on PDA spreading, with sparse aerial mycelium and crenate margins; surface pale mouse-grey, outer region grey-olivaceous; reverse grey-olivaceous, reaching up to 25 mm diam after 1 mo; on MEA spreading, erumpent with sparse aerial mycelium; surface prominently striate, margin crenate; centre black, outer region mouse-grey; reverse black; colonies reaching up to 20 mm diam after 1 mo.

Specimens examined. Brazil, São Paulo Horto Botanico, leaves of Spiraea cantoniensis, Sept. 1905, leg. Usteri no. 15 bis, holotype LPS 12280; Rio Grande do Sul, Guaiba, living leaves of Stenocalyx uniflorus, 1 Apr. 2008, leg. A.C. Alfenas, isol. P.W. Crous, epitype designated here CBS H-20206, culture ex-type CPC 15151 = CBS 123533, CPC 15152.

Notes — The holotype specimen (LPS 12280) was described and illustrated in detail by Nag Raj (1977). The species was originally described from leaves of Spiraea cantoniensis collected in the São Paulo Botanical Garden, where it occurred on leaves of several tree species, suggesting that it is not host specific. The present collection was obtained by incubating Eugenia leaves with leaf spots of Phaeophleospora eugeniae in moist chambers, which resulted in a few conidiophores of Vonarxia vegans developing.

Nag Raj (1977) erected Kazulia for a genus of hyphomycetes with dark brown, septate setae, and tetraradiate conidia, which he regarded as morphologically distinct, and a probable anamorph of the Chaetothyriaceae. The fact that he did not compare Kazulia with Vonarxia is not surprising, because Batista et al. (1960) who initially described Vonarxia, showed setae on the outside of the pycnidia, and thus this fungus was regarded as a coelomycete. Later comments from Nag Raj (1977) (as Kazulia) suggest, however, that these bodies are perithecia of a probable teleomorph. In a subsequent study Van der Aa & Van Oorschot (1985) and Van der Aa & Von Arx (1986) showed that Kazulia is a synonym of Vonarxia. Wu & Sutton (1995) were not convinced of the distinction between Vonarxia and another hyphomycete genus, Fumagopsis, due to insufficient material, and chose to use the name Fumagopsis for F. complexa, which they described from Eugenia leaves collected in India. Based on the present collection of V. vagans, it is apparent, that these are two distinct genera. In Fumagopsis the setae are aseptate, arranged around the sporodochium, and taxa have rhizoid-like structures. In contrast, the setae of Vonarxia are septate, irregularly distributed and do not surround the sporodochium, and have a simple, bulbous base.

Setae 87–155 μm long; apical conidial arms 12–35 μm long ………V. anacardii

Setae and conidial arms longer; setae 120–220 μm long; apical conidial arms 20–55(–90) μm long…… V. vagans

Crous, Mycol. Mem. 21: 154. 1998

Type species. Xenostigmina zilleri (A. Funk) Crous.

Associated with leaf spots. Mycelium internal, consisting of hyaline to pale brown, septate, branched, smooth hyphae. Conidiomata sporodochial, brown to black. Conidiophores densely aggregated, arising from the upper cells of a pale brown stroma, finely verruculose, hyaline to pale brown, multi-septate, subcylindrical, straight to variously curved, branched. Conidiogenous cells terminal and intercalary, hyaline to pale brown, finely verruculose, doliiform to subcylindrical, tapering to flat tipped loci, proliferating sympodially and percurrent; loci not thickened or conspicuous. Conidia solitary, pale to medium brown, with pale brown apical and basal regions, finely verruculose, mostly straight, ellipsoidal, apex subobtuse, frequently extending into a beak; base truncate at dehiscence, inner part extending later to form a short, subobtuse basal appendage; septation muriform; basal marginal frill present.

(A. Funk) Crous, Mycol. Mem. 21: 155. 1998 — Fig 15

Fig. 15

Xenostigmina zilleri (CBS 124108). a–c. Conidial propagules of Mycopappus aceris; d. setae on the surface of conidial propagules; e. colony of Xenostigmina zilleri; f, g. fasciculate conidiophores; h. conidia. — Scale bars = 10 μm.

Basionym. Stigmina zilleri A. Funk, Canad. J. Bot. 65: 482. 1987.

Synanamorph. Mycopappus aceris (Dearn. & Barthol.) Redhead & G.P. White, Canad. J. Bot. 63: 1430. 1985.

Basionym. Cercosporella aceris Dearn. & Barthol., Mycologia 9: 362. 1917.

Teleomorph. ? Didymella mycopappi (A. Funk & Dorworth) Crous, Mycol. Mem. 21: 152. 1998.

Basionym. Mycosphaerella mycopappi A. Funk & Dorworth, Canad. J. Bot. 66: 295. 1988.

Characteristics in culture — Colonies spreading on PDA with moderate to abundant aerial mycelium, and feathery margins; olivaceous-grey with patches of iron-grey and pale olivaceous-grey; iron-grey in reverse. On OA spreading, with abundant aerial mycelium, olivaceous-grey with patches of pale olivaceous-grey. On MEA erumpent, spreading, with abundant aerial mycelium, pale olivaceous-grey with patches of olivaceous-grey and iron-grey; reverse iron-grey.

Specimens examined. Canada, British Columbia, 15 km east of Sardis, on living leaves of Acer macrophyllum, 22 Oct. 1985, A. Funk & C.E. Dorworth, holotype DAVFP 23272; British Columbia, on living leaves of Acer sp., 2002, leg. K.A. Seifert, isol. P.W. Crous, CBS 115686 = CPC 4010, CBS 115685 = CPC 4011; Victoria BC, 48°30′25.63″N, 123°30′46.99″W, 115 m, fallen leaves of Acer macrophyllum, 6 Sept. 2007, leg. B. Callan, isol. P.W. Crous, CBS H-20208, CPC 14376 = CBS 124108, CPC 14377, 14378 (Xenostigmina zilleri), CPC 14379 = CBS 124109, CPC 14380, 14381 (Mycopappus aceris).

Notes — Although Stigmina s.str. has been shown to reside in Pseudocercospora s.str. (Crous et al. 2006a, Braun & Crous 2006, 2007), this is not the case for Xenostigmina (Crous 1998), which appears to be related to Seifertia (Seifert et al. 2007) in the Dothideomycetes. Isolates of the Xenostigmina state are shown here (Fig. 1) to be identical to those of the Mycopappus state, which proves that these two genera are indeed synanamorphs. No ascospore isolates were obtained, however, to confirm their relationship to ‘Mycosphaerella’ mycopappi, though this species is clearly not a member of the Mycosphaerellaceae. Xenostigmina wolfii (Crous & Corlett 1998), which is the anamorph of Mycosphaerella stigmina-platani, and a Pseudocercospora synanamorph, is not congeneric with X. zilleri, and would be better accommodated in Pseudocercospora (Crous et al. 2006a) than in Xenostigmina.