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STUDIES IN MYCOLOGY 50: 95–108. 2004.
95
Conioscyphascus, a new ascomycetous genus for holomorphs with
Conioscypha anamorphs
Martina Réblová1* and Keith A. Seifert2
1Department of Plant Taxonomy and Biosystematics, Institute of Botany, Academy of Sciences, 252 43 Průhonice, Czech
Republic; 2Biodiversity (Mycology and Botany), Agriculture and Agri-Food Canada, Environment Theme, Ottawa, Ontario,
K1A 0C6, Canada
*Correspondence: Martina Réblová, reblova@ibot.cas.cz
Abstract: The new genus Conioscyphascus is described for the teleomorph of the hyphomycete Conioscypha varia, and a
second species, C. gracilis, based on Debaryella gracilis. Species of the new genus produce inconspicuous, superficial or
immersed, ostiolate, smooth, subhyaline to pale orange perithecia with two-layered walls, hyaline, septate paraphyses, 8-
spored unitunicate asci with a refractive J– apical annulus, and fusiform 3−7-septate ascospores. Phylogenetic analyses of
alignments of large- and small-subunit rDNA sequences suggest a strongly supported, close relationship among the three
Conioscypha species sampled and Ascotaiwania and Carpoligna, but the family and order relationships of this clade remain
uncertain.
Taxonomic novelties: Conioscyphascus Réblová & Seifert gen. nov., Conioscyphascus varius Réblová & Seifert sp. nov.,
Conioscyphascus gracilis (Munk) Réblová & Seifert comb. nov.
Key words: Cryptoleptosphaeria, Debaryella, Diaporthales, Glomerellaceae, life cycles, LSU and SSU rDNA, phylogeny.
INTRODUCTION
Conioscypha Höhn. is a dematiaceous hyphomycete
genus with eight terrestrial and freshwater species
inhabiting decayed wood, leaves, or bamboo stems;
these species have also been isolated from skin scrap-
ings and hair of living animals (Shearer 1973, Matsu-
shima 1975, Udagawa & Toyazaki 1983, Kirk 1984,
Matsushima 1993, 1996, Chen & Tzean 2000). The
genus is characterized by an unusual mode of conidio-
genesis that includes aspects of both phialidic and
annellidic ontogeny (Shearer 1973, Shearer & Motta
1973, Cole & Samson 1979, Goh & Hyde 1998).
Conidiogenesis occurs at inconspicuous loci along
hyphae; a basipetal succession of blastically produced
conidia leave behind conspicuous collarettes that are
remnants of the initial outer wall of the conidia; these
accumulate centripetally to form a multi-layered
collarette appearing similar to annellations (Goh &
Hyde 1998). To our knowledge, the ultrastructure of
the process has not been studied and there is some
conjecture as to whether these conidiogenous cells are
really phialides, and how the collarettes are produced.
The phylogenetic relationships of Conioscypha are
unknown and no teleomorph connections have been
reported.
On specimens of decorticated, strongly decayed
wood collected in central Europe, we encountered a
nonstromatic, perithecial ascomycete that produced
small, pale orange perithecia with an elongate neck
and several-layered wall, unitunicate, long-stipitate
asci with a J– apical annulus, and fusiform, septate,
hyaline ascospores. No anamorph was noted in vivo,
but isolated ascospores produced a Conioscypha
anamorph, apparently identical with C. varia Shearer
(1973).
The teleomorph of this fungus matches the descrip-
tion of the genus Debaryella Höhn. (Höhnel 1904),
erected for the single species D. hyalina Höhn., a
parasite with ascomata that develop in perithecial
cavities of the lignicolous, stromatic fungus Eutypa
scabrosa (Bull.) Fuckel. A second species, Debaryella
vexans Höhn., a parasite on stromata of wood-
inhabiting species of Valsa Fr. or Diaporthe Nitschke,
was added later to the genus and is now considered a
synonym of Cryptonectriella biparasitica (Höhn.)
Weese in the Diaporthales (Rossman et al. 1999).
Munk (1957) suggested that Debaryella was di-
aporthaceous and described a third species, the sapro-
bic, lignicolous fungus D. gracilis Munk. Subsequent
references to Debaryella in the literature are based
primarily on the original description of D. hyalina
(Rogerson 1970, Barr 1978, Samuels 1988). Unfortu-
nately, the type material of D. hyalina is depauperate
(Kohlmeyer et al. 1997, Rossman et al. 1999) and the
protologue lacks diagnostic features that are critical
for its assignment to a family or order. Therefore,
Kohlmeyer et al. (1997), Rossman et al. (1999) and
Eriksson (2000) considered Debaryella a nomen
dubium, but the name could be resurrected if suitable
material is found to serve as an epitype.
RÉBLOVA & SEIFERT
96
Table 1. List of substrates, localities, sources, relevant sexual and asexual states and accession numbers of taxa sequenced in this study.
Teleomorph Anamorph Locality and substrate Sourcea GenBank
LSU SSU
Conioscyphascus
varius
Conioscypha varia Czech Republic, decayed deciduous wood CBS 113653 AY484512 AY484511
– Conioscypha japonica* Japan, scrapings and hair of dog CBS 387.84 AY484514 –
– Conioscypha lignicola* Australia, dead leaf base of Xanthorrhoea
preissii
CBS 335.93 AY484513 –
Chaetosphaeria
curvispora
Chloridium-like New Zealand, decayed decorticated wood CBS 113644 – AY502933
*Sequences of conidial isolate. aCBS = Centraalbureau voor Schimmelcultures, Utrecht, the Netherlands.
Cryptoleptosphaeria Petr., originally described as a
monotypic genus in the Hypocreales (Petrak 1923), is
also relevant to this discussion. The generic diagnosis
of Cryptoleptosphaeria is similar to that of Debary-
ella, based on D. hyalina, i.e. hyaline, thin-walled
ascomata, unitunicate cylindrical asci with J– apical
annulus, and long-ellipsoidal to fusiform, septate,
hyaline ascospores; the species have a similar ecology.
Cryptoleptosphaeria moravica Petr., the type species,
occurs in locular cavities of a foliicolous, Leptosphae-
ria-like ascomycete. Rossman et al. (1999) classified
Cryptoleptosphaeria in the Diaporthales and consid-
ered it a relative of Debaryella, transferring D.
gracilis to the genus as C. gracilis (Munk) Rossman &
Samuels. On the basis of its type material (Czech
Republic, Moravia: Hranice na Moravě, Skalická near
Bečva, in the ascomata of a Leptosphaeria-like fun-
gus, on Phalaris arundinacea; FH, isotype), C. mo-
ravica is not congeneric with C. gracilis, differing by
its thin, single-layered ascomatal wall, disintegrating
paraphyses, asci with an obtuse base that lacks a stipe,
and by its parasitic habitat. However, C. gracilis is
morphologically similar to our undescribed fungus
with the Conioscypha anamorph, differing only by its
smaller asci and ascospores, hyaline to subhyaline,
short-papillate perithecia and smaller conidia. A
different Conioscypha anamorph with smaller conidia,
attached to the surface of the perithecia and nearby
woody substrate, was also found on the type and other
herbarium material of C. gracilis preserved at C
herbarium.
Cryptoleptosphaeria gracilis and our undescribed
fungus with the Conioscypha varia anamorph also
superficially resemble the foliicolous saprobic genus
Juncigena Kohlm. et al., which includes the single
species J. adarca Kohlm. et al. (Magnaporthaceae;
Kohlmeyer et al. 1997). Juncigena differs from the
teleomorphs of Conioscypha by its brownish asco-
mata, a centrum with pseudoparaphyses attached at
both the bottom and the top of the ascomatal cavity,
short-stipitate asci and a helicosporous hyphomycete
anamorph, classified in Cirrenalia Goos.
Despite the characteristic teleomorphs and ana-
morphs, we were unable to assign Cryptoleptosphae-
ria gracilis and the undescribed fungus with the
Conioscypha anamorph to any known genus of non-
stromatic, perithecial ascomycetes or an ascomycetous
order or family. In this study, we introduce the new
genus Conioscyphascus for holomorphs with Conio-
scypha anamorphs; Debaryella (Cryptoleptosphaeria)
gracilis is transferred to this new genus. A key and
taxonomic descriptions for the two accepted species
are provided, and we explore their systematic position
and phylogenetic relationships using phylogenetic
analyses of nuclear small (SSU rDNA) and large
subunit ribosomal DNA (LSU rDNA).
MATERIALS AND METHODS
Herbarium material and cultures
Dried herbarium specimens were rehydrated in 3 %
(aq.) KOH and studied in water, Melzer’s reagent or
90 % lactic acid. All measurements were made in
lactic acid. Means ± standard errors (se) are given for
spore and ascus dimensions and are based on 20–25
measurements. Images were captured in Melzer’s
reagent using differential interference microscopy
(DIC) and phase contrast (PC) and processed using
Adobe Photoshop 6.0 CE.
Single-ascospore isolates were obtained from fresh
material with the aid of a single-spore isolator
(Meopta). Cultures were grown on potato-carrot agar
(PCA, Gams et al. 1998). Colony characters were
taken from cultures grown on PCA for 14 d at room
temperature (about 25 °C) in incident light. Colour
codes in descriptions refer to Kornerup & Wanscher
(1978).
Cultures are maintained at the Institute of Botany,
Academy of Sciences in Průhonice, the Canadian
Collection of Fungal Cultures (DAOM) and the Cen-
traalbureau voor Schimmelcultures (CBS). Strains
used in this study and their sources are listed in Table
1.
DNA extraction, amplification and sequencing
Methods for DNA extraction, amplification and se-
quencing of the first two thirds of the LSU (1600 bp,
though only 1260 bp were used for the analysis, in
order to match the length of other sequences) were
identical to those described by Réblová & Seifert
(2004).
For amplification and sequencing of the SSU,
methods were similar to those of Hambleton et al.
CONIOSCYPHASCUS GEN. NOV
97
(2003), except that UltraClean Microbial DNA Isola-
tion and UltraClean PCR Purification kits (Mo Bio
Laboratories Inc., Solana Beach, CA) were used for
DNA extraction and cleaning of PCR products, and an
ABI PRISM® 3700 DNA Analyzer (Applied Biosys-
tems, Foster City CA) was used for sequencing.
Sequence data analyses
Phylogenetic relationships were examined using 49
LSU rDNA and 106 SSU rDNA sequences retrieved
from GenBank, representing 8 or 9 orders of ascomy-
cetes; accession numbers are given on Figs 1–3.
Members of the Dothideomycetes, Taphrinomycetes
and Saccharomycetes were used as outgroups in the
separate maximum parsimony analyses of the LSU
and SSU rDNA. The data sets included new LSU and
SSU sequences of the fungus described below as
Conioscyphascus varius (ascospore isolate), Conio-
scypha japonica Udagawa & Toyazaki and C. ligni-
cola Höhn. (conidial isolates) and Chaetosphaeria
curvispora Réblová & Seifert (SSU only).
All sequences were manually aligned in BioEdit
5.0.9 (Hall 1999). Predicted models of the secondary
structure of the LSU (Gutell et al. 1993) and SSU
(Gutell 1993) rRNA molecules of Saccharomyces
cerevisiae were used to improve the alignment (HEB
alignment, Eriksson 2000). The models of the secon-
dary structure of the LSU and SSU rRNA were highly
consistent in all taxa and were comparable with that of
Saccharomyces cerevisiae. The alignments are avail-
able in TreeBase as M2046.
Phylogenetic analyses were performed with PAUP
4.0b10 (Swofford 2002) using maximum parsimony
(heuristic search with 100 random sequence addi-
tions). All analyses were run with gaps treated as
missing data. Support for the branches was tested with
1000 replicates of bootstrap and jackknife analyses
using fast step-wise addition. Constraint analyses were
run using the Kishino-Hasegawa test as implemented
in PAUP, with the Diaporthales, Hypocreales, Mi-
croascales, Glomerella clade, Plectosphae-
rella/Verticillium clade, Conioscyphascus, or various
interpretations of these groups forced to be distinct
and monophyletic, as explained below.
RESULTS
Phylogenetic analysis of the LSU rDNA sequence
data
A first maximum parsimony analysis (PA1) was
performed using 386 phylogenetically informative
characters in an alignment including 1260 bp from 50
taxa. Two most parsimonious trees (MPTs) were
obtained, one of them shown in Fig. 1. The trees
differed only in the arrangement of branches for
Lanatonectria flavolanata. The Xylariales (79 %
bootstrap support / 78 % jackknife support), Di-
aporthales (98/97), Microascales (99/100), Ophio-
stomatales (99/98), Magnaporthaceae (100/100) and
the Glomerella clade (89/90) were well-supported
clades at the order or family levels. The Chaetosphae-
riales was strongly supported (99/99). The Sordaria-
les appeared as two major discontinuous lineages
separated by the Chaetosphaeriales; one lineage
(65/63) including the Chaetomiaceae (Sordariales,
chaetomiaceous complex sensu Huhndorf et al. 2004)
and the Sordariaceae with the poorly supported
Lasiosphaeriaceae (Sordariales, lasiosphaeriaceous
complex sensu Huhndorf et al. 2004) as a second
lineage. The Hypocreales were poorly supported
(65/65). Conioscyphascus and Conioscypha were in a
well-supported clade of ambiguous ordinal affiliation,
here called the Conioscyphascus/Carpoligna/Asco-
taiwania clade (95/95, abbreviated as CCA clade),
which was basal to all sampled unitunicate ascomy-
cetes. Conioscyphascus (including the two other
Conioscypha species) (88/86) and two species of
Ascotaiwania Sivan. & H.S. Chang (with so-called
Monotosporella S. Hughes anamorphs) (99/99), were
sister groups and strongly supported subclades within
the clade. Carpoligna pleurothecii was basal to the
Conioscyphascus/Conioscypha clade, but without
bootstrap/jackknife support.
LSU sequences of Ascotaiwania hughesii Fallah et
al. (with a Helicoon Morgan anamorph) and A. per-
soonii Fallah et al. were excluded from PA1 because
they required insertions in the stem parts of the secon-
dary structure to align them with the remainder of the
data set. A second analysis, PA2, was performed on a
reduced data set to assess the phylogenetic relation-
ships of these two additional Ascotaiwania species,
including only parts of the sequences that could be
unambiguously aligned without disrupting the secon-
dary structure. PA2 included 324 phylogenetically
informative characters in an alignment of 1270 bp
including 30 taxa (including representatives of the
Hypocreales, Microascales, Glomerella clade, Chae-
tosphaeriales, the CCA clade, and the Xylariales with
the Dothideales as outgroup). Two MPTs were ob-
tained with similar topologies and support to PA1.
One of them is shown in Fig. 2. The Chaetosphaeria-
les and the CCA clade were sister groups but with no
bootstrap/jackknife support. Within the CCA clade
(90/90) (Fig. 2), the Conioscyphascus/Conioscypha
clade (83/74) and Ascotaiwania/Monotosporella clade
(100/100) were monophyletic and well-supported.
Ascotaiwania hughesii and A. persoonii were para-
phyletic with the other Ascotaiwania spp. at the base
of the clade, although the taxa were on a single branch
with no support.
Phylogenetic analysis of the SSU rDNA sequence
data
A third maximum parsimony analysis (PA3) was
performed with 567 phylogenetically informative
RÉBLOVA & SEIFERT
98
characters in an alignment of 1781 bp including 108
taxa (tree length 2815, CI = 0.338, RI = 0.725, HI =
0.662). Forty six MPTs (not shown) were obtained,
with Taphrinomycetes and Saccharomycetes as out-
groups. Seven ascomycetous classes were included,
i.e. Pezizomycetes, Leotiomycetes, Sordariomycetes,
Dothideomycetes, Lecanoromycetes, Chaetothyriomy-
cetes and Eurotiomycetes. Conioscyphascus formed a
monophyletic clade within the Sordariomycetes
(99/100) with Plectosphaerella cucumerina (Lindf.)
W. Gams, Verticillium dahliae and Volutella colleto-
trichoides J.E. Chilton (in a similar arrangement to
Fig. 1). This clade was placed within a larger clade
including the monophyletic Glomerella clade and its
two sisters, the Microascales and the Hypocreales.
Therefore, in subsequent analyses we focused only on
the class Sordariomycetes (including the same sam-
pling of 38 taxa), with representatives of the Do-
thideomycetes chosen as outgroup.
A maximum parsimony analysis (PA4) was per-
formed using the 319 phylogenetically informative
characters in an alignment of 1781 bp including 44
taxa. Two MPTs were obtained, which differed only
in the grouping of taxa within the Xylariales. One of
the 2 MPTs is shown in Fig. 3. The well-supported
clades were again the Diaporthales (100/100), Mi-
croascales (85/84) and the Glomerella clade (97/93).
In this analysis, the Sordariales (94/93) and Chae-
tosphaeriales (100/100) obtained high support. A
large monophyletic clade (78/75) contained mono-
phyletic Hypocreales (54/54) and Microascales
(85/84) on one branch, and monophyletic Glomerella
clade and a clade (84/82) including P. cucumerina, V.
dahliae and V. colletotrichoides (100/100) and Co-
nioscyphascus on a second branch. This was a sister
group to the clade with Sordariales, Diaporthales and
Xylariales within the Sordariomycetes.
Constraint analyses
The results of the LSU and SSU rDNA analyses
provided apparently contradictory hypotheses con-
cerning the sister group and family/order placement of
Conioscyphascus. In the LSU analysis, the closest
relatives to Conioscyphascus were Carpoligna pleu-
rothecii and two species of Ascotaiwania that could
not be included in the SSU analysis. These taxa were
on a long branch basal to the rest of the unitunicate
pyrenomycetes. In the SSU analysis, the closest
relatives of Conioscyphascus were Plectosphaerella
cucumerina, and representatives of the Verticillium
dahliae group, which together were a sister group to
the Glomerella clade. In other words, when Car-
poligna and Ascotaiwania were present in the data set,
they pulled Conioscyphascus away from the
Glomerella clade and the Plectosphae-
rella/Verticillium clade. To determine whether the
SSU and LSU analyses were in conflict concerning
the relationships of Conioscyphascus, we ran several
constraint analyses (CA) using monophylies suggested
by the SSU to test the robustness of the LSU clades,
and vice versa.
For the LSU rDNA data set, four CAs were run to
test the possible monophyly of the Conioscyphas-
cus/Carpoligna/Ascotaiwania clade with P. cucu-
merina and V. dahliae (CA1); P. cucumerina, V.
dahliae and the Glomerella clade (CA2); P. cucu-
merina and V. dahliae with Ascotaiwania removed
(CA3); and P. cucumerina, V. dahliae and the
Glomerella clade with Ascotaiwania removed (CA4).
In CA1, the resulting three trees (not shown) were 12
steps longer and were not considered statistically
different from the MPT by the Kishino-Hasegawa
(KH) test (P* = 0.3373, 0.3310 and 0.3213). CA2
resulted in two trees (not shown) 10 steps longer than
the two MPTs and were not rejected by the KH test
(P* = 0.3918 and 0.3620). However, for CA3 and
CA4, with the two Ascotaiwania species with Mono-
tosporella anamorphs excluded from the Conioscy-
phascus/Carpoligna group, the resulting nine and two
constraint trees (not shown) were 69 and 63 steps
longer; all were rejected by the KH test (P* = 0.0001).
Two further CAs were run to test the possible rela-
tionship to the Diaporthales of Conioscyphascus alone
(CA5) and to the Conioscyphascus/Ascotaiwania
clade (CA6), following previous suggestions by Munk
(1957) and Rosmann et al. (1999) for C. gracilis. In
CA5 and CA6, the five and seven constraint trees (not
shown) were 64 and 73 steps longer than the two
MPTs; all were rejected by the KH test (P* = 0.0001).
In the SSU rDNA data set, two CAs were run to
assess the inclusion of Conioscyphascus, the
Glomerella clade and the Plectosphae-
rella/Verticillium clade in the Hypocreales (CA7), or
the Hypocreales and Microascales (CA8). In CA7, the
six trees (not shown) were seven steps longer than the
MPT and the KH test did not reject them (P* =
0.4070, 0.3709, 0.3787, 0.3933, 0.3460, 0.3460). In
CA8, the fifteen constraint trees (not shown) were 10
steps longer and were also considered acceptable by
the KH test (P* values ranged from 0.1492 to 0.2189).
CONIOSCYPHASCUS GEN. NOV
99
Setosphaeria monoceras AY016368
Pleospora herbarum AF382386
Dothidea sambuci AF382387
Chaetomium globosum U47825
Neurospora crassa M28154
Sordaria fimicola AF132330
Codinaea simplex AF178559*
Chaetosphaeria innumera AF178551
Cercophora newfieldiana AF064642
Lasiosphaeria ovina AF064643
Diaporthe padi AF408354
Diaporthe pustulata AF408357
Valsa ambiens AF362564
Leucostoma niveum AF408367
Cryptodiaporthe aesculi AF408342
Apiognomonia errabunda AF408334
Gnomonia gnomon AF408361
Phragmoporthe conformis AF408377
Cryptosporella hypodermia AF408347
Melanconis stilbostoma AF362567*
Xylaria hypoxylon U47841
Graphostroma platystoma AY083827
Diatrype disciformis U47829
Daldinia concentrica U47828
Cainia graminis AF431949
Amphisphaeria umbrina AF452029
Lepteutypa cupresii AF382379
Oxydothis frondicola AY083835
Magnaporthe grisea AB026819
Gaeumannomyces graminis AF362557
Ophiostoma piliferum AY281094
Ophiostoma africanum AF221015
Petriella sordida AY281099
Petriella setifera AY281100
Microascus trigonosporus U47835
Hypocrea schweinitzii AY281095
Hypomyces subiculosus AY281096
Lanatonectria flavolanata AY281098
Glomerella phacidiomorpha AF275496
Colletotrichum nymphaeae AY705727*
Glomerella cingulata U17403
Colletotrichum gloeosporioides AY705727*
Plectosphaerella cucumerina U17399
Verticillium dahliae U17425*
Ascotaiwania mitriformis AF132324
Ascotaiwania sawadae AF132323
Conioscypha japonica AY484514*
Conioscypha lignicola AY484513*
Conioscyphascus varius AY484512
Carpoligna pleurothecii AF064646
10 changes
99/99
74/73
99/98
100/100
99/100
84/84
65/65
100/100
95/94
55/56
53/50
89/87
100/100
70/70
98/97
57/59
81/80
79/78
100/100
86/82
88/86
99/99
95/95
96/94
100/100
89/90
100/100
75/75
100/100
100/100
65/63
DOTHIDEOMYCETES
Magnaporthaceae
DIATRYPALES/
XYLARIALES
DIAPORTHALES
OPHIOSTOMATALES
MICROASCALES
HYPOCREALES
Glomerella clade
Conioscyphascus/
Carpoligna/Ascotaiwania clade
SORDARIALES pro parte
Plectosphaerella/Verticillium clade
SORDARIALES pro parte
CHAETOSPHAERIALES
Fig. 1. One of two equally parsimonious trees from a heuristic analysis (PA1) of LSU rDNA sequences from 7 ascomycetous
orders. Bootstrap and jackknife values (> 50 %) from 1000 replicates are included at the nodes. The asterisk (*) indicates taxa
represented by the anamorph in the phylogeny. Branch lengths are drawn to scale (1821 steps, CI = 0.383, RI = 0.683, HI =
0.617).
Fig. 2. Part of one of two equally parsimonious trees from a
heuristic analysis (PA2) of LSU rDNA sequences from 7
ascomycetous orders, showing the Conioscyphas-
cus/Carpoligna/Ascotaiwania clade (CCA clade). Bootstrap
and jackknife values (> 50 %) from 1000 replicates are
included at the nodes. The asterisk (*) indicates taxa repre-
sented by the anamorph in the phylogeny. Branch lengths
are drawn to scale (1225, CI = 0.442, RI = 0.664, HI =
0.558).
RÉBLOVA & SEIFERT
100
Cochliobolus sativus U42479
Setosphaeria rostrata U42487
Pleospora betae U43466
Melanomma sanguinarium AF242265
Lophiostoma crenatum U42485
Rhytidhysteron rufulum AF201452
Lasiosphaeria ovina AY083799
Madurella mycetomatis AF527811*
Chaetomium elatum M83257
Sordaria fimicola X69851
Phyllachora graminis AF064051
Chaetosphaeria curvispora AY502933
Kionochaeta ramifera AB003788*
Camarops microspora Z49783
Coniochaeta ligniaria AJ496242
Gnomonia setacea AF277121
Gnomoniella fraxini AF277106
Xylaria carpophila Z49785
Hypoxylon fragiforme AB014046
Microdochium nivale AF548077*
Hyponectria buxi AF130976
Graphostroma platystoma AY083808
Pestalosphaeria hansenii AF242846
Apiospora sinensis AY083815
Arthrinium phaeospermum AY083816*
Diatrype disciformis U32403
Gibberella pulicaris AF081467
Chaetopsina fulva B003786*
Hypocrea lutea D14407
Cordyceps ophioglossoides AB113827
Sphaerodothis acrocomiae U76340
Nectria pseudotrichia AY342011
Microascus cirrosus M89994
Petriella setifera U43908
Pseudallescheria boydii U43915
Ceratocystis fimbriata U43777
Graphium penicillioides AB038423*
Colletotrichum gloe osporiodes M55640*
Glomerella cingulata M55640
Glomerella septospora U78779
Plectosphaerella cucumerina AF176951
Verticillium dahliae U33637*
Volutella colletotrichoides AJ301962*
Conioscyphascus varius AY 484511
10 changes
92/93
100/100
87/88
76/70
76/72
94/93
100/100
61/60
68/6564/63
100/100
86/81
54/54
53/52
100/100
97/93
80/81
100/100
84/82
72/73
92/91
85/84
59/61
98/98
100/99
66/63
78/75
100/100
SORDARIALES
DOTHIDEOMYCETES
DIAPORTHALES
DIATRYPALES/
XYLARIALES
MICROASCALES
HYPOCREALES
SORDARIOMYCETES
Glomerella clade
PHYLLACHORALES
CHAETOSPHAERIALES
CONIOCHAETALES
Plectosphaerella/
Verticillium clade
BOLINIALES
Fig. 3. One of two equally parsimonious trees from a heuristic analysis (PA4) of SSU rDNA sequences from 6 ascomycetous
orders. Bootstrap and jackknife values (> 50 %) from 1000 replicates are included at the nodes. The asterisk (*) indicates taxa
represented by the anamorph in the phylogeny. Branch lengths are drawn to scale (912 steps, CI = 0.464, RI = 0.738, HI =
0.536).
TAXONOMY
The undescribed perithecial teleomorph of Conioscy-
pha varia and two other species, C. japonica and C.
lignicola, the type species of the genus, formed a well-
supported, monophyletic clade in all our phylogenetic
analyses. This clade could not be assigned unambigu-
ously to a known order or family, and constraint
analyses suggested the Hypocreales, Microascales,
Glomerellaceae or the Plectosphaerella/Verticillium
clade as equally possible closely related taxa. Fur-
thermore, no particular relationship with members of
the Diaporthales was evident. Because neither mature
authenticated material nor other herbarium material of
Debaryella hyalina are available to confirm the iden-
tity and taxonomic position of Debaryella, the new
genus Conioscyphascus is proposed for holomorphs
with Conioscypha anamorphs. One new species is
described and D. gracilis is transferred to the new
genus.
Conioscyphascus Réblová & Seifert, gen. nov.
MycoBank MB500024.
Anamorph: Conioscypha Höhn., Ann. Mycol. 2: 58.
1904. emend. Shearer, Mycologia 65: 128. 1973.
Etymology: Conioscypha, the generic name of the
anamorph; -ascus (Gk) refer to the part of the organ-
ism that reproduces sexually.
Ascomata immersa vel superficialia, singula, conica vel
globosa vel subglobosa, subhyalina vel armeniaca, papillata
usque ostiolo centrali elongato, cylindraceo, praedita.
Paraphyses copiosae, septatae, hyalinae. Paries perithecii
coriaceus, bistratosus. Asci cylindraceo-clavati, unitunicati,
8-spori. Ascosporae fusiformes usque naviculares, septatae,
non constrictae, hyalinae.
Anamorphe Conioscypha. Conidiophora micronemato-
sa, mononematosa. Cellulae conidiogenae hyalinae, inter-
calares vel ex apice hypharum ortae; conidia liberata
collaria multiplicia relinquentia. Conidia brunnea, continua.
Species typica: Conioscyphascus varius Réblová & Seifert
sp. nov.
Perithecia superficial or immersed, solitary, glabrous,
hyaline to subhyaline to pale orange, globose to sub-
globose with subcylindrical papilla or cylindrical
elongated neck, ostiole periphysate. Perithecial wall
leathery, waxy, two-layered, cells in the outer wall ca.
CONIOSCYPHASCUS GEN. NOV
101
2–2.5 µm diam, thick-walled. Paraphyses present,
hyaline, septate, partly disintegrating at maturity,
broader at the base, tapering towards the tip, apically
free, longer than the asci, arising from the ascogenous
hyphae. Asci unitunicate, cylindrical-clavate, stipitate,
apical annulus distinct, refractive, J–. Ascospores
fusiform to fusiform-navicular, hyaline, 3–7-septate,
smooth-walled. Conidiophores micronematous,
mononematous. Conidiogenous cells terminal or
intercalary, hyaline, cyathiform to doliiform, with
multilayered, hyaline, cup-like collarettes. Conidia
formed singly and successively by percurrent prolif-
eration of the conidiogenous cell and seceding by
apical rupture of the outer wall. Conidia variable in
shape, 1-celled, dark brown, sometimes with a central
pore at the base.
Type species: Conioscyphascus varius Réblová &
Seifert sp. nov.
Commentary: Conioscyphascus is distinguished from
other perithecial ascomycetes by a unique combina-
tion of characters, including subhyaline to pale orange
perithecia, unitunicate, stipitate, cylindrical-clavate
asci with a distinct, J– apical annulus, apically free
paraphyses, fusiform, septate, hyaline ascospores, and
its Conioscypha anamorphs.
The perithecia of Conioscyphascus species are
subhyaline to pale-coloured, very inconspicuous and
scarcely visible on the natural substratum after drying.
This might explain the few reports of these fungi
(Munk 1957, Minoura & Muroi 1978, Romero 1994).
In both specimens of C. varius, we saw evagina-
tion of the apical ring outside the apical part of the
ascus at maturity, and the ascospores were released
through it (Figs 6, 7, 24c). In material of Debaryella
gracilis, Munk (1957: fig. 72d) observed that the
distal part of the ascus broke off along a circular zone
to liberate the ascospores. Our study of the type and
other material of D. gracilis from Munk’s herbarium
(C) did not confirm this observation. The apical ring
of D. gracilis has the same structure as the ring of C.
varius and seems to be fully functional, although
evagination was not observed.
The Conioscypha varia anamorph obtained in vitro
from both collections of C. varius frequently produced
conidia with basal arms (Figs 12, 13). Normally,
delimitation of the conidia occurs at the base of the
collarette, allowing continued production of conidia.
In some conidiogenous apertures, however, delimita-
tion of the conidia occurs in the subtending hypha,
effectively locking the conidium into place and pre-
venting any further formation of conidia from that
locus (W. Gams, pers. comm.).
The central basal pore of the conidia (Fig. 16) may
function during germination, but we did not observe
conidia of C. varia germinating in culture. Similar
pores were also reported for C. japonica and C. tai-
waniana J.L. Chen & S.S. Tzean (Chen & Tzean
2000). The pore on natural material of C. gracilis was
eccentric, occurring on one side of the conidium at the
base.
Key to species of Conioscyphascus [For a key to Conioscypha species refer to Goh & Hyde (1998).]
1. Perithecia pale orange, long-beaked; ascospores longer than 35 µm; asci 120–160 µm long in
pars sporifera ............................................................................................................................................. C. varius
1. Perithecia subhyaline or dirty wood-colour, short papillate; ascospores usually up to 36 µm
long; asci 85–120(–150) µm long in pars sporifera................................................................................. C. gracilis
Conioscyphascus varius Réblová & Seifert, sp.
nov. MycoBank MB500025. Figs 4–18, 26, 27.
Anamorph: Conioscypha varia Shearer, Mycologia
65: 133. 1973.
= Cylicogone regenerans Emden & Veenb.-Rijks,
Acta Bot. Neerl. 22: 637. 1973.
Ascomata collo excepto immersa, singula, globosa vel
subglobosa, armeniaca, 350–450 µm diam, 350–460 µm
alta, ostiolo centrali elongato, cylindraceo 300–350 µm
longo praedita. Paraphyses copiosae, hyalinae. Asci
cylindraceo-clavati, 120–160 (mean ± se = 146.5 ± 5) µm
longi in parte sporifera, 10–12(–13) (mean ± se = 11.5 ±
0.2) µm lati, stipite (40–)52–80(–95) µm longo, 8-spori.
Ascosporae fusiformes usque naviculares, (34–)36–41(–
42) (mean ± se = 38.4 ± 0.4) × (3.5–)4–4.5(–5) (mean ± se
= 4 ± 0.3) µm, 5–7-septatae, non constrictae, hyalinae.
Anamorphe Conioscypha varia. Conidiophora
micronematosa, mononematosa. Cellulae conidiogenae
hyalinae, intercalares vel ex apice hypharum ortae collis
multiplicibus, 11–18 (mean ± se = 14.8 ± 1) µm longis,
(6–)8–10 (mean ± se = 8.3 ± 0.5) µm latis. Conidia
ellipsoidea, brunnea, ad basim truncata, 10–18(–19)
(mean ± se = 14.5 ± 0.7 × 6–10(–11) (mean ± se = 8 ± 0.4)
µm, continua, laevia.
RÉBLOVA & SEIFERT
102
Figs 4–25. Conioscyphascus spp. 4–18. Conioscyphascus varius. 4. Ascus. 5, 6. Ascospores. 7, 8. Apical annulus (arrows
indicate evagination of annulus). 9–18. Conioscypha varia anamorph, conidia and conidiogenous cells (arrows indicate: 13,
14, 17 conidial arms; 16 basal circular pore), from culture on PCA. 19–25. Conioscyphascus gracilis. 19. Conidium of the
Conioscypha sp. anamorph, from nature. 20, 21. Asci. 22–24. Ascospores. 25. Paraphyses. DIC: 4–6, 8–11, 13–24; PC: 7, 12,
25. Conioscyphascus varius: 4–7 from PRM 900537 (holotype), 8 from PRM 901091; 9–11, 13–15, 17, 18 from CBS 113653;
12, 16 from CBS 113654. Conioscyphascus gracilis: 20, 22, 23, 25 from C 24673; 21, 24 from C (holotype). Scale bars: Figs 4,
20, 21 = 20 µm; 5–18, 22–25 = 10 µm.
Perithecia completely immersed with only the neck
protruding above the surface, solitary, pale orange,
CONIOSCYPHASCUS GEN. NOV
103
indistinct, collapsing laterally upon drying, globose to
subglobose, 350–450 µm diam, 350–460 µm high,
glabrous, neck 300–350 µm high, 110–120 µm wide,
central, cylindrical, upright. Perithecial wall (27–)37–
50 µm thick; outer wall of thick-walled, brick-like
cells that become thinner towards the interior, inner
layer of thin-walled, hyaline, elongated cells; wall in
the neck formed of slightly diverging rows of elon-
gated and thick-walled cells, textura porrecta to
epidermoidea in surface view. Paraphyses abundant,
hyaline, septate, 4–6 µm wide near the base, tapering
to 2–2.5 µm, partly disintegrating at maturity. Asci
cylindrical-clavate, 120–160 (mean ± se = 146.5 ± 5)
µm long in pars sporifera, 10–12(–13) (mean ± se =
11.5 ± 0.2) µm wide, long-stipitate, stipe (40–)52–
80(–95) µm long, L/W 12.8:1 (for asci in pars sporif-
era), truncate to broadly rounded at the apex, apical
annulus refractive, J–, distinct, evaginate at maturity
outside the ascus, with ascospores discharged through
the evaginated ring. Ascospores fusiform-navicular,
tapering towards the ends and slightly curved, (34–)
36–41(–42) (mean ± se = 38.4 ± 0.4) × (3.5–)4–4.5
(–5) (mean ± se = 4 ± 0.3) µm, L/W 9.6:1, hyaline,
narrowly rounded at both ends, 5–7-septate, not con-
stricted a the septa, smooth-walled.
Characteristics in culture: Colony 12–13 mm diam,
whitish to pale yellowish, having a slimy appearance,
with a marginal greyish ring because of the dark
brown conidia. Mycelium usually immersed in the
substrate, aerial mycelium scarcely developed and
tightly appressed to the surface of the colony; hyphae
hyaline, 2–2.5 µm wide, septate, smooth. Sporulation
copious, at first limited to a marginal ring, after 1–2
mo sporulation widespread throughout the colony.
Margin entire, discrete. Reverse whitish to pale yel-
lowish. Conidiophores micronematous, mononema-
tous, arising terminally or laterally, hyaline, reduced
to a minute protuberance on the hyphae. Conidioge-
nous cells terminal or intercalary, hyaline, cyathiform,
with multilayered, hyaline, cup-like collarette 11–18
(mean ± se = 14.8 ± 1) µm long, (6–)8–10 (mean ± se
= 8.3 ± 0.5) µm wide in the flaring distal part, com-
posed of layers of previously ruptured outer walls of
conidiogenous cell. Conidia ellipsoidal, slightly
truncate at the base, apically rounded, 10–18(–19)
(mean ± se = 14.5 ± 0.7) µm long, 6–10(–11) (mean ±
se = 8 ± 0.4) µm wide, L/W 1.8:1, basal scar 3–4 µm
wide, conidia frequently with basal arms, dull brown,
aseptate, smooth-walled, with a central, round pore at
the base, formed singly and successively by percurrent
proliferation of the apex of the conidiogenous cell,
separating by apical rupture of the outer wall of the
conidiogenous cell. Chlamydospores abundant, termi-
nal or intercalary on hyphae, ellipsoidal to obpyriform
to irregularly shaped, 6–17 (mean ± se = 12.3 ± 1.1)
µm long, 6–10 (mean ± se = 7.4 ± 0.4) µm wide,
aseptate, dull brown, smooth-walled.
Known distribution: Czech Republic, U.S.A. (Mary-
land).
Habitat: Saprobic on decayed wood.
Holotype: Czech Republic, Southern Bohemia, Šumava
Mts. National Park, Povydří National Nature Reserve,
Modrava, brook Zhůřský potok close to Turner’s hut,
decayed deciduous wood, 27 Oct. 2001, M. Réblová 1890-
01 (PRM 900537, holotype).
Additional specimen examined: Czech Republic, Southern
Bohemia, Šumava Mts. National Park, Povydří National
Nature Reserve, Čeňkova Pila, decayed wood of Ulmus
glabra, 27 Aug. 2000, M. Réblová 1676-00 (PRM 901091).
Cultures: CBS 113653 (ex-holotype PRM 900537), CBS
113654 (ex-PRM 901091).
Commentary: The anamorph of C. varius formed in
vitro is identical to Conioscypha varia, originally
isolated from submerged balsa wood. In the type
culture of C. varia (Shearer 1973), the conidia varied
greatly in shape from ovoidal to flame-shaped to
navicular to subellipsoidal and were slightly shorter
than those of our strain.
RÉBLOVA & SEIFERT
104
Fig. 26. A–C. Conioscyphascus varius. A. Ascus with paraphyses. B. Ascospores. C. Ascal apex with apical annulus during the
discharge of ascospore. D–F. Conioscyphascus gracilis. D. Ascus with paraphyses. E. Ascospores. F. Conidia of Conioscypha
anamorph, from nature. A–C from PRM 900537 (holotype); D–F from C (holotype). Scale bars = 10 µm.
Conioscyphascus gracilis (Munk) Réblová &
Seifert, comb. nov. MycoBank MB500026. Figs
19–25, 26, 28.
≡ Debaryella gracilis Munk, Bot. Tidsskr. 51: 226.
1957.
≡ Cryptoleptosphaeria gracilis (Munk)
Rossman & Samuels, Stud. Mycol. 42: 185.
1999.
Anamorph: Conioscypha sp.
Perithecia usually superficial, with the base slightly
immersed, rarely completely immersed, solitary,
subhyaline to pale yellowish brown to dirty wood-
colour, indistinct, collapsing laterally upon drying,
subglobose, 170–200 µm diam, 180–250 µm high,
glabrous, papilla subcylindrical, broadly rounded to
obtuse at the apex. Perithecial wall 42–50 µm thick,
outer wall of yellowish, thick-walled, polyhedral cells
that become more elongated towards the interior;
inner layer of thin-walled, hyaline, elongated cells;
wall in the papilla formed of slightly diverging rows
of elongated, thick-walled cells, textura porrecta in
surface view. Paraphyses abundant, hyaline, filiform,
septate, 2.5–3 µm wide near the base, tapering to 1.5–
2 µm, longer than the asci. Asci cylindrical-clavate,
85–120(–150) (mean ± se = 111 ± 2.7) µm long in
pars sporifera, 7–10 (mean ± se = 11 ± 0.2) µm wide,
stipe 12–27 µm long, L/W 10:1, truncate at the apex,
CONIOSCYPHASCUS GEN. NOV
105
apical annulus refractive, J–, 3–3.5 µm diam, 1.5–2
µm high. Ascospores fusiform, (24–) 26–36(–37)
(mean ± se = 30.4 ± 0.6) × (3.5–)4–5 (mean ± se = 4.4
± 0.06) µm, L/W 7:1, hyaline, straight or slightly
curved, narrowly rounded at both ends, 3–5-septate,
not constricted at the septa, smooth-walled.
Fig. 27. Conioscyphascus varius. A. Median, longitudinal
section of perithecium. B. Habit sketch of perithecia. From
PRM 900537 (holotype). Scale bar = 100 µm.
Anamorph: Several conidia of a putative Conioscypha
anamorph were attached to the outer wall of the
perithecia. Conidia ellipsoidal, slightly truncate at the
base, apically rounded and slightly tapering, 8–10
(mean ± se = 9 ± 0.4) µm long, 6–7 (mean ± se = 6.4
± 0.4) µm wide, L/W 1.3:1, basal scar ca. 3 µm wide,
dull brown with a pore near the base. Mycelium or
conidiogenous cells not observed.
Illustrations and descriptions: Munk (1957: 194, fig.
72), Shearer (1973: 128, figs 3, 7a–d), Minoura &
Muroi (1978: 129, fig. 2), Romero (1994: 76, fig. 8a–
c, plate IX–1, 2).
Known distribution: Argentina, Denmark, Japan.
Habitat: Freshwater and terrestrial saprobe on decayed
wood.
Holotype: Denmark, Lunden park in Silkeborg, decayed
wood of a stump (?Viburnum lantana), associated with
Nectria sp., 2 Apr. 1954, A. Munk (C).
Additional specimens examined: Denmark, Sjælland,
Bernstorffsparken, decayed wood of decorticated branch,
23 Mar. 1965, A. Munk 15 (C); Sjælland, Tokkenkøb Hegn,
decayed decorticated wood, 20 Jun. 1993, T. Læssøe (C
32138); North Jylland, Høstemark Skov, decorticated wood
of twigs and branches of Ribes rubrum/nigrum, associated
with Nectria sp., 10 Apr. 1995, T. Læssøe (C 24673).
Fig. 28. Conioscyphascus gracilis. A. Median, longitudinal
section of perithecium. B. Habit sketch of perithecia. From
C (holotype). Scale bar = 100 µm.
Commentary: The conidial dimensions of the putative
anamorph of C. gracilis do not match any of the
presently known species of Conioscypha. The asci in
the type collection of C. gracilis were somewhat
larger (115–150 µm in pars spori-fera) than the asci
of other specimens examined (85–106 µm in pars
sporifera). Larger asci were reported also by Romero
(1994, 120–140 µm for the whole ascus) on material
from Argentina. Munk (1957) did not report larger
asci in the type and reported an ascus size of 80–95
µm in pars sporifera.
RÉBLOVA & SEIFERT
106
DISCUSSION
The phylogenetic affinities of the monophyletic clade
including Conioscyphascus, Ascotaiwania and Car-
poligna (CCA clade) to the other sampled unitunicate
ascomycetes remain unclear and the high degree of
DNA divergence between the Conioscyphascus clade
and the other taxa included in this study was unex-
pected. Despite different grouping of Conioscyphas-
cus in the LSU and SSU rDNA analyses, the con-
straint analyses testing the monophyly of
Conioscyphascus with the Hypocreales, Microascales,
Glomerellaceae and the Plectosphaerella clade,
resulted in trees that were accepted by the KH test as
possible alternative phylogenetic hypotheses.
There are a few common morphological features
that unite the genera of the CCA clade: thin-walled
unitunicate asci with a distinct, J– apical annulus; true,
apically free paraphyses; symmetrical, transversely
septate ascospores; absence of stromatic tissue or
clypeus and similar anatomies of the perithecial walls.
The genera are distinguished by the hyaline asco-
spores of Conioscyphascus and Carpoligna versus the
concolorous (pale brown) or versicolorous ascospores
(brown middle cells, hyaline polar cells) of Ascotai-
wania. The colour of the perithecial wall also differs,
being hyaline or pale orange in Conioscyphascus and
dark or opaque in Ascotaiwania and Carpoligna. The
anamorphs of these genera represent two different
modes of conidiogenesis. The Conioscypha ana-
morphs of Conioscyphascus species exhibit a unique
mode of conidiogenesis with multiple, conspicuous
collarettes forming a multilamellar structure around
the blastic (presumed to be phialidic) conidiogenous
locus. The other species exhibit variations of ho-
loblastic conidiogenesis. The Pleurothecium recurva-
tum (Höhn.) Morgan anamorph of Carpoligna pleu-
rothecii F.A. Fern. et al. (Fernández et al. 1999) and
the Helicoon farinosum Linder anamorph of Ascotai-
wania hughesii Fallah et al. (Fallah et al. 1999) have
rhexolytic conidial secession on denticulate, sympo-
dially proliferating conidiogenous cells. The so-called
Monotosporella anamorphs of Ascotaiwania mitri-
formis (Ranghoo & Hyde 1998) and A. sawadae
(Sivichai et al. 1998) have a single terminal conidio-
genous locus or lateral loci lacking denticles. These
latter phragmoconidial anamorphs are aleurioco-
nidium-like, and borne singly on erect, monoblastic
conidiogenous cells. They are a poor fit in Monoto-
sporella, which otherwise includes species with per-
currently proliferating conidiogenous cells, and in-
cludes presumed teleomorphs in the Hysteriaceae
(Hughes 1979). Acarocybiopsis J. Mena et al. (1999)
may be a better fit for these anamorphs, although there
are still deviating characters.
Fernández et al. (1999) discussed the possible
affinities of Carpoligna F.A. Fern. & Huhndorf with
the Hypocreales and Microascales on the basis of
partial LSU rDNA sequence data. Carpoligna has
relatively few, rather simple morphological characters,
which makes its recognition as a distinct taxon diffi-
cult in absence of knowledge of its anamorph. It
mimics Chaetosphaeria Tul. & C. Tul. in particular,
but the Pleurothecium Höhn. anamorph differs sig-
nificantly from the phialidic anamorphs accepted in
Chaetosphaeria (Réblová & Winka 2000, Réblová
2000).
The structure of the apical annulus of Conioscy-
phascus species is reminiscent of the annulus of some
members of the Diaporthales. However, a similarly
well-developed annulus also occurs in species of
Ascotaiwania and Carpoligna, which are sister groups
to Conioscyphascus in the phylogenetic analyses.
Based on the presently available LSU and SSU rDNA
sequence data, Conioscyphascus has obviously no
affinity to the Diaporthales (Figs 1, 3). Members of
the Diaporthales usually have short-stipitate asci that
float free, the centrum is aparaphysate or has paraphy-
soid tissues that deliquesce early in development, and
conidiogenesis is enteroblastic. Rossman et al. (1999)
placed Cryptoleptosphaeria in the Diaporthales based
on features of the asci and the apical annulus of the
type species of C. moravica. Our transfer of C.
gracilis to Conioscyphascus leaves Cryptoleptosphae-
ria monotypic; its morphology is fully documented by
Rossman et al. (1999) based on the type material of C.
moravica.
Ceratosphaeria Niessl, based on C. lampadophora
(Berk. & Broome) Niessl, shares hyaline, long-
fusiform, septate ascospores with Conioscyphascus, as
well as unitunicate asci with a well-developed, J–
apical annulus, true, apically free paraphyses, and
globose to subglobose perithecia with elongated
central necks occurring on strongly decayed wood. It
differs by its perithecial wall, composed of the darkly
pigmented cells with opaque walls, the Harpophora-
like anamorph, and its phylogenetic affinity (LSU
rDNA) with members of the Magnaporthaceae (Ré-
blová, unpublished).
Presently, Ascotaiwania is considered “incertae
sedis” on the basis of LSU rDNA analyses (Ranghoo
et al. 1999, Réblová & Winka 2001). The genus was
classified in the Amphisphaeriaceae sensu lato by
Sivanesan & Chang (1992). Our analysis (Fig. 1) does
not support the relationship of Ascotaiwania with the
Amphisphaeriaceae (Xylariales), represented in our
LSU rDNA analysis by Amphisphaeria umbrina (Fr.)
De Not., Lepteutypa cupressi (Nattrass et al.) H.J.
Swart and Oxydothis frondicola K.D. Hyde. The
apical annulus of the ascus of Ascotaiwania species
was later used as a critical feature to accommodate the
genus in the Annulatascaceae (Wong et al. 1998), a
family erected mainly on the basis of a single feature,
i.e. a large apical annulus. According to our LSU
rDNA analyses, the Annulatascaceae are polyphyletic,
with the core species having affinities with the
CONIOSCYPHASCUS GEN. NOV
107
Trichosphaeriales (Réblová & Seifert 2004). Al-
though the Monotosporella-like and Helicoon ana-
morphs of the three Ascotaiwania species discussed
above were obtained in vitro, the anamorphs of eight
other Ascotaiwania species remain unknown (Sivane-
san & Chang 1992, Hyde 1995, Chang et al. 1998,
Fallah et al. 1999, Hyde & Goh 1999, Dulymamode et
al. 2001, Wong & Hyde 2001). The occurrence of two
morphologically distinctive types of conidia, conidio-
phores and conidiogenesis in Ascotaiwania suggest
that the genus might not be monophyletic as currently
circumscribed. However, our parsimony analysis (Fig.
2) supported a relationship of A. hughesii (with a
Helicoon anamorph) and A. persoonii (anamorph
unknown) with the Ascotaiwania/Monotosporella
subclade (100/100), although they were basal to that
group and Ascotaiwania was then paraphyletic with
the other taxa in the clade. Sequences of the type
species, A. lignicola Sivan & H.S. Chang, are unavail-
able.
It is difficult to find an appropriate group among
existing ascomycete orders to accommodate Conio-
scyphascus, Ascotaiwania and Carpoligna. Because of
differences in teleomorph morphologies and in the
mode of conidiogenesis associated with certain types
of peridial morphology, especially the degree of
pigmentation, we can only hypothesize that these three
genera may finally represent distinct groups at the
family or order level. However, such a hypothesis
needs to be tested with sequence data for additional
taxa.
ACKNOWLEDGEMENTS
This project was supported by the Grant Agency of Acad-
emy of Sciences (GAAV B6005106, KSK6005114), the
Research Project of the Institute of Botany, Academy of
Sciences of the Czech Republic (AV0Z6005908) and
COBICE project to M.R. (May 2001). We thank the follow-
ing herbaria for their loan of the type and other material: C,
FH. We are grateful to Dr W. Gams for comparison of our
Conioscypha varia strains with his own material and for
confirming its identity. Dr Sarah Hambleton facilitated our
SSU analysis by providing aligned sequences from her data.
Presubmission reviews were provided by Bob Shoemaker
and Sarah Hambleton.
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