Molecular phylogeny of Boliniales (Sordariomycetes) with an assessment of the systematics of Apiorhynchostoma, Endoxyla and Pseudovalsaria.
ABSTRACT The systematics of ascomycete genera Apiorhynchostoma, Endoxyla and Pseudovalsaria are re-evaluated based on the comparison of cultural characteristics, teleomorph morphology and DNA sequence data. Analyses of internal transcribed spacer (ITS) of the ribosomal DNA operon and large subunit (LSU) of the nuclear ribosomal DNA gene sequences resolve Boliniales as a robustly supported lineage comprising Apiorhynchostoma, Camarops, Camaropella, Cornipulvina, Endoxyla and Pseudovalsaria. Within Boliniales, species of Endoxyla form a strongly supported lineage. Apiorhynchostoma curreyi and Pseudovalsaria ferruginea group with Cornipulvina ellipsoides. Species of Camarops are paraphyletic and comprise two clades, one of which includes Camaropella. Boliniaceae is emended, Endoxyla mallochii is described as new and Apiorhynchostoma trabicola is considered a synonym of Apiorhynchostoma altipetum. We also propose the combinations Endoxyla occulta, Endoxylina luteobasis and Jobellisia peckii. Keys to genera included in the Boliniaceae and to species of Apiocamarops, Apiorhynchostoma and Endoxyla are provided.
- Boliniaceae transferred to Sordariales (Ascomycota). 1-16..
- Synopsis Pyrenomycetum (continued from p 86). 122-125..
- Clypeosphaeria and the Clypeosphaeriaceae Prodromus to nonlichenized, pyrenomyce-tous members of the class Hymenoascomycetes Redisposition of some taxa described by Notes on the Amphisphaeriaceae and related families. 1-843..
Molecular phylogeny of Boliniales (Sordariomycetes) with an assessment
of the systematics of Apiorhynchostoma, Endoxyla and Pseudovalsaria
Wendy A. Untereiner1
Department of Biology, Brandon University, Brandon,
Manitoba, R7A 6A9 Canada
PO Box 1137, Cornwall, PEI, C0A 1H0 Canada
H.W. (Bud) Platt
Agriculture and Agri-Food, Charlottetown, PEI, C1A
Birkagatan 49, SE-256 55 Helsingborg, Sweden
Department of Biology, Natural History Museum of
Denmark, University of Copenhagen, DK-1353
Va ´clav Sˇte ˇpa ´nek
Laboratory of Enzyme Technology, Institute of
Microbiology, Academy of Sciences, 142 20 Prague,
Martina Re ´blova ´
Department of Taxonomy, Institute of Botany, Academy
of Sciences, 252 43 Pru ˚honice, Czech Republic
Apiorhynchostoma, Endoxyla and Pseudovalsaria are re-
evaluated based on the comparison of cultural
characteristics, teleomorph morphology and DNA
sequence data. Analyses of sequences of the internal
transcribed spacer (ITS) of the ribosomal DNA
operon and the large subunit (LSU) of the nuclear
ribosomal DNA gene resolve Boliniales as a robustly
supported lineage comprising Apiorhynchostoma, Ca-
marops, Camaropella, Cornipulvina, Endoxyla and
Pseudovalsaria. Within Boliniales, species of Endoxyla
form a strongly supported lineage. Apiorhynchostoma
curreyi and Pseudovalsaria ferruginea group with
Cornipulvina ellipsoides. Species of Camarops are
paraphyletic and comprise two clades, one of which
includes Camaropella. Boliniaceae is emended, En-
doxyla mallochii is described as new and Apiorhynch-
ostoma trabicola is considered a synonym of Apior-
hynchostoma altipetum. We also propose the
combinations Endoxyla occulta, Endoxylina luteobasis
and Jobellisia peckii. Keys to genera included in the
The systematics of the ascomycete genera
Boliniaceae and to species of Apiocamarops, Apior-
hynchostoma and Endoxyla are provided.
Boliniaceae, hyaline cap, infrequently
collected microfungi, lignicolous pyrenomycetes, po-
rate ascospores, systematics
Species of Apiorhynchostoma Petrak and Endoxyla
Fuckel are morphologically similar ascomycetes
found primarily on the wood of gymnosperms in
central and northern Europe and North America
(Untereiner 1993, Rogers et al. 1994b). Members of
both genera possess immersed, perithecial ascomata
with stout necks, unitunicate asci with an inamyloid
apical ring and brown porate ascospores (Mu ¨ller and
von Arx 1962, Untereiner 1993). Their stromata are
usually poorly developed and evident only as dark-
ened or discolored wood around or between the
ascomata (Untereiner 1993, Rappaz 1995); in some
species the stromata are described as clypeate
(Sivanesan 1975, Re ´blova ´ 1998, Wang et al. 2004).
Ascospores of species of Apiorhynchostoma are two-
septate and consist of two dark brown cells and a
small, terminal hyaline cell. The brown cell farthest
from the hyaline cell is smaller and possesses a
terminal germ pore (Mu ¨ller and von Arx 1962) that is
covered by an indistinct hyaline cap or appendage
(Sivanesan 1975, Rappaz 1995). Species of Endoxyla
possess brown, porate ascospores that are septate or
aseptate (Untereiner 1993), and a hyaline cap does
not cover the porate end of the ascospore.
Apiorhynchostoma and Endoxyla are considered
closely related (Untereiner 1993, Rogers et al.
1994b, Re ´blova ´ 1998), but their taxonomic positions
have remained uncertain. Both genera have been
included in the Clypeosphaeriaceae and have been
compared to Clypeosphaeria Fuckel (Barr 1989, 1990;
Untereiner 1993; Rogers et al. 1994b; Re ´blova ´ 1998;
Wang et al. 2004). Apiorhynchostoma and Endoxyla
also have been treated as members of the Boliniales
(Rappaz 1995, Huhndorf and Miller 2008), an order
inferred as closely related to the Chaetosphaeriales
and Sordariales based on the phylogenetic analysis of
four loci (Zhang et al. 2006).
Determining the phylogenetic positions of Apior-
hynchostoma and Endoxyla has proven challenging
because these species are collected infrequently.
Endoxyla macrostoma Fuckel, the type species of the
Submitted 11 Sep 2012; accepted for publication 8 Nov 2012.
1Corresponding author. E-mail: email@example.com
Mycologia, 105(3), 2013, pp. 564–588. DOI: 10.3852/12-326
# 2013 by The Mycological Society of America, Lawrence, KS 66044-8897
genus, has been reported from only three collections
outside Fuckel’s herbarium (Untereiner 1993, Re ´-
blova ´ 1998, Johnova ´ 2009), while A. altipetum (Peck)
Rappaz, A. occultum Re ´blova ´, A. trabicola You Z.
Wang, Aptroot & K.D. Hyde., A. tumulatum (Cooke)
Sivan. and E. xanthostroma Unter. are known from
two or fewer collections (Sivanesan 1975, Untereiner
1993, Rappaz 1995, Re ´blova ´ 1998, Wang et al. 2004,
Johnova ´ 2009). Attempts to obtain cultures of species
of Apiorhynchostoma and Endoxyla have proven
unsuccessful (Untereiner 1993, Re ´blova ´ 1998); as a
consequence, the members of these genera are
unrepresented in culture collections and in molecu-
lar phylogenetic studies.
In 2007 we cultured Apiorhynchostoma curreyi
(Rabenh.) E. Mu ¨ller from material collected in
Sweden. Shortly thereafter we were able to culture
Pseudovalsaria ferruginea (Nitschke) Rappaz and a
number of species of Endoxyla collected in Canada.
One collection of Endoxyla represented an unde-
scribed species. Prompted by the availability of
cultures and fresh collections of these taxa, we
obtained nuclear ribosomal gene sequences (rDNA)
and examined the phylogeny of these genera and
their closest relatives. Based on the examination of
recent collections and existing herbarium material,
we describe this new species in Endoxyla, propose the
new combination in Endoxyla for Apiorhynchostoma
occultum, reduce A. trabicola to synonymy with A.
altipetum and accept a single species, P. ferruginea, in
Pseudovalsaria Spooner. We also propose the combi-
nation in Endoxylina Romell for Sphaeria luteobasis
Ellis and transfer Valsa peckii Howe to Jobellisia M.E.
Barr. Our analyses of rDNA gene sequences confirm
the placement of Apiorhynchostoma, Endoxyla and
Pseudovalsaria in Boliniales.
MATERIALS AND METHODS
Fungal strains and cultural studies.—The isolates examined
in this study (TABLE I) were obtained from freshly collected,
air-dried collections by streaking the contents of mature
ascomata on modified Leonian’s agar (MLA) (Malloch
1981) containing 2.5% agar, chlortetracycline (50 mg/mL)
and streptomycin sulfate (50 mg/mL). Individual or groups
of germinating ascospores were subcultured to MLA
containing 1.5% agar and maintained at room temperature
(20–21 C). Ascomata of species of Endoxyla also were
streaked on MLA amended with the chemical adsorbents
1% bovine serum albumin (BSA, Sigma A-7030, Sigma-
Aldrich, Oakville, Ontario) or 1% a-cyclodextrin (Sigma
C-4642) (Denison 2003).
For comparative purposes, cultures were grown on
filtered oatmeal agar (CBSOA) (Gams et al. 1987),
cornmeal agar (CMA) (Gams et al. 1987) using only 30 g
ground cornmeal, 2% malt extract agar (MEA), MLA and
oatmeal agar (OA) (Tuite 1969). Plates were inoculated in
triplicate with 2 3 2 mm squares cut from the actively
growing edges of colonies on MLA and incubated at room
temperature. Colony diameter was measured and descrip-
tions of colony morphology were made at 7 d intervals for
21 d. Color descriptions are based on Kornerup and
Wanscher (1978). Unless noted, the color of the reverse
of colonies was the same as the colony surface.
Microscopic examinations of cultures and specimens
were made from preparations mounted in distilled water,
lactophenol containing 0.1% cotton blue (Kirk et al. 2001)
or Melzer’s reagent (Malloch 1981). Asci and ascospores
also were observed from preparations mounted in 1%
aqueous Janus green B (JGB) following removal of stain in
distilled water. Freehand sections of the ascomata and
substrate were examined to determine the extent of
stromatal development. The acronyms of herbaria are those
used by Holmgren et al. (1990).
Extraction, amplification and sequencing of DNA.—Cultures
used for DNA isolations were grown as described in
TABLE I. Sources and accession numbers of the isolates sequenced in this study
TaxonSubstrate and geographic originSourcesa
GenBank accession nos.
Decorticated wood of
Pinus sylvestris L., Sweden
Decorticated wood of
Pinus strobus L., Canada
Decorticated wood (Abies sp.
or Picea sp.), Canada
Decorticated wood of Populus
cf. balsamifera L., Canada
Decorticated wood of Populus sp.,
C, UAMH 11088JX460984JX460989
NBM, UAMH 11491JX460985JX460990
NBM, UAMH 11087JX460986 JX460991
NBM, UAMH 11085JX460987JX460992
NBM, UAMH 11490JX460988JX460993
aVouchers are deposited in the University of Copenhagen Herbarium (C), Copenhagen, Denmark, or in the New Brunswick
Museum (NBM), St John, Canada. Cultures are deposited in the University of Alberta Microfungus Collection and Herbarium
(UAMH), Edmonton, Canada.
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
Untereiner et al. (1995) and Re ´blova ´ et al. (2011). Total
nucleic acids were extracted from mycelia following the
protocols of Lee and Taylor (1990) or Re ´blova ´ et al. (2011).
Methods for amplifying and sequencing the internal
transcribed spacer (ITS) and the large subunit (LSU) of
the nuclear rDNA gene are described in Bogale et al. (2010)
and Re ´blova ´ et al. (2011). Primers used in the amplification
and sequencing of the ITS included ITS4, ITS5, NS5 (White
et al. 1990), LR0R (Rehner and Samuels 1994), LR1 and
5.8SR (Vilgalys and Hester 1990, Hopple and Vilgalys 1999);
primers used to amplify and sequence the LSU included
LR5, LR6, LR7, LR8, LR16 (Vilgalys and Hester 1990,
Hopple and Vilgalys 1999), JS7 and JS8 (Landvik 1996).
Sequence alignment and phylogenetic analyses.—Newly gen-
erated sequences were compared to homologous sequences
retrieved from GenBank, primarily from studies of Re ´blova ´
et al. (2004), Spatafora et al. (2006), Zhang et al. (2006),
Huhndorf and Miller (2008) and Re ´blova ´ et al. (2011).
Sequences were manually aligned in BioEdit 220.127.116.11 (Hall
1999). The LSU alignment was enhanced with the
homologous 2D structure of Saccharomyces cerevisiae Meyen
ex E.C. Hansen (Gutell et al. 1993). All characters in the ITS
alignment were included in phylogenetic analyses, but we
excluded bp 1–73 from the LSU dataset because of the
incompleteness of the 59– end of the majority of the
available sequences. The combined ITS and LSU dataset
was partitioned into two subsets, ITS (ITS1-5.8S-ITS2) and
LSU. Alignments are deposited in TreeBASE (accession
The positions of Apiorhynchostoma, Endoxyla and Pseudo-
valsaria within Boliniales (Sordariomycetidae) were in-
ferred based on the analysis of the LSU sequences of 77
taxa representing 13 orders or families of Sordariomycetes.
Saccharomyces cerevisiae and Vanderwaltozyma polyspora (van
der Walt) Kurtzman (Saccharomycetes) were used as
outgroup taxa. We also analyzed the combined ITS and
LSU of sequences of 12 taxa belonging to Boliniales,
Chaetosphaeriales and Sordariales. Outgroup taxa for these
analyses included Chaetosphaeria innumera Berk. & Broome
ex Tul. & C. Tul., C. pulviscula (Curr.) C. Booth and C.
vermicularioides (Sacc. & Roum.) W. Gams & Hol.-Jech.
MrModeltest 2.3. (Nylander 2008) was used to infer the
appropriate substitution model that would best fit the
model of DNA evolution for our datasets. Maximum
likelihood (ML) and Bayesian inference (BI) analyses were
used to estimate phylogenetic relationships. ML analysis was
performed with RAxML-HPC 7.0.3 (Stamatakis 2006) using
GTR + I + G (LSU datasets) and GTR + G (ITS dataset) models
of evolution. Nodal support was verified by nonparametric
bootstrapping (BS) with 1000 replicates. BI analysis was
performed in a likelihood framework as implemented by
MrBayes 3.0b4 (Huelsenbeck and Ronquist 2001). Multiple
Bayesian searches were conducted using Metropolis-cou-
pled Markov chain Monte Carlo sampling. One cold and
three heated Markov chains were conducted. Analyses were
run 5000000 generations, with trees sampled every 1000
generations. The first 20000 trees, representing the burn-in
phase of the analysis, were discarded. To estimate posterior
probabilities (PP) of recovered branches (Larget and Simon
1999), 50% majority rule consensus trees were created from
the remaining trees with PAUP* 4.0b 10 (Swofford 2003).
Ascospores from fresh, air-dried collections of Apior-
hynchostoma curreyi, Endoxyla macrostoma, E. opercu-
lata (Fr.: Fr.) Sacc. and Pseudovalsaria ferruginea
germinated within 36–72 h on MLA by forming single
germ tubes that emerged through the germ pore (not
shown). Macroscopically visible colonies were evident
on MLA within 5 d. The ascospores of the unnamed
species of Endoxyla were never observed to germinate
on MLA or on MLA containing BSA and a-cyclodex-
trin, but slow growing, orange-white colonies origi-
nated from the remnants of ascomata on all media.
All species produced flat, white or yellow to orange
colonies that consisted largely or entirely of an
immersed mycelium. Conidial anamorphs were not
observed on any of the media used.
The LSU dataset for 77 taxa contained 1286
characters of which 395 were phylogenetically infor-
mative. The ML phylogram and a detailed view of a
robustly supported clade (ML bootstrap 5 95%, PP 5
1.0) within the subclass Sordariomycetidae containing
Boliniales, Chaetosphaeriales, Coniochaetales and
Sordariales are illustrated (FIG. 1). ML analysis of
these data (mean likelihood 5 212912; transitions A–
C 5 0.925, A–G 5 2.845, A–T 5 1.725, C–G 5 0.833,
C–T 5 7.890) resolved Boliniales as a strongly
supported clade (81/0.98) that was sister to Chaeto-
sphaeriales (FIG. 1). Within Boliniales, species of
Endoxyla comprised a monophyletic group (50/
0.75) that was sister to a lineage comprising two
subclades. The first subclade (61/0.97) included
Apiorhynchostoma curreyi, Cornipulvina ellipsoides
Huhndorf, A.N. Mill., F.A. Ferna ´ndez & Lodge and
Pseudovalsaria ferruginea; the second subclade (53/
0.98) consisted of five species of Camarops (C. petersii
[Berk. & M.A. Curtis] Nannf., C. polysperma [Mont.]
J.H. Mill., C. rogersii Huhndorf & Miller, C. tubulina
[Alb. & Schwein.] Shear and C. ustulinoides [Henn.]
Nannf.). Camarops amorpha (Boedijn) Nannf. and C.
microspora (P. Karst.) Shear grouped with species of
Camaropella Lar.N. Vassiljeva in a lineage (53/0.94)
that was sister to the clade encompassing Apiorhynch-
ostoma, Cornipulvina Huhndorf, A.N. Mill., F.A.
Ferna ´ndez & Lodge, Endoxyla, Pseudovalsaria and
five other species of Camarops.
The combined ITS and LSU dataset for 12 taxa
consisted of 1808 characters; 263 of these were
phylogenetically informative. In the ML tree (mean
likelihood 5 26690; ITS partition transitions A–C 5
0.620, A–G 5 12.054, A–T 5 14.472, C–G 5 10.398,
C–T 5 37.101; LSU partition transitions A–C 5 0.456,
Phylogram inferred from ML analysis of LSU rDNA sequences. Part A depicts the entire phylogeny whereas part B shows in detail the robustly supported clade
containing Boliniales, Chaetosphaeriales, Coniochaetales and Sordariales. ML bootstrap and BI posterior probabilities are indicated at the nodes. Branches receiving
significant support (ML BS 5 100%, PP 5 1.0) are indicated by an oval.
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
A–G 5 1.278, A–T 5 0.876, C–G 5 0.384, C–T 5
5.566), the Boliniales was resolved as a robust clade
(100/1.0) containing two subclades (FIG. 2). Within
the first subclade (96/0.98), species of Endoxyla
formed a strongly supported monophyletic group
(99/1.0) that was sister to a lineage that included
Camarops polysperma and C. ustulinoides (100/1.0).
The second subclade (2/0.74) included Apiorhynch-
ostoma curreyi and Pseudovalsaria ferruginea.
Boliniaceae, although traditionally allied with the
Xylariales (Munk 1953, Barr 1990, Romero and
Samuels 1991), was shown by Andersson et al.
(1995) to be more closely related to Sordariales based
on the analysis of small subunit ribosomal DNA
sequences. The family, together with Catabotryda-
ceae, was placed subsequently in Boliniales (Kirk et al.
2001). Recent molecular phylogenies demonstrate
that the closest relatives of Boliniaceae within
Sordariomycetes include Chaetosphaeriales, Conio-
chaetales and Sordariales (Huhndorf et al. 2004,
Miller and Huhndorf 2005, Zhang et al. 2006).
Catabotrydaceae is a monotypic family based on
Catabotrys decidua (Berk. & Broome) Seaver &
Waterson, a widespread species found on the dead
leaves and petioles of tropical monocots that forms
long-necked, perithecial ascomata in superficial,
pulvinate stromata, and ellipsoid, hyaline to lightly
pigmented ascospores (Seaver and Waterson 1946,
Petrak 1954, Barr 1990). Although it is morphologi-
cally similar to some Boliniaceae, C. decidua is not
closely allied to this family and was excluded from
Boliniales (Huhndorf et al. 2004). Analysis of a
combined b-tubulin-LSU-RPB2 dataset indicates that
C. decidua is a member of Diaporthales (Miller and
Huhndorf 2005), but the description of the apical
ring of the asci of C. decidua as amyloid (Dingley
1977) supports the earlier suggestion (Hyde et al.
2000) that this species might belong to Xylariales.
Our analyses of LSU sequence data resolve
Apiorhynchostoma, Camaropella, Camarops, Cornipul-
vina, Endoxyla and Pseudovalsaria as members of
Boliniales (FIG. 1) and are consistent with molecular
phylogenies that include this order as a strongly
supported lineage within Sordariomycetes (Huhndorf
et al. 2004, Miller and Huhndorf 2005, Zhang et al.
probabilities are indicated at the nodes. GenBank accession numbers given in the tree after the names are those of ITS/LSU.
Branches receiving significant support (ML BS 5 100%, PP 5 1.0) are indicated by an oval.
Phylogram inferred from ML analysis of a combined data ITS-LSU rDNA sequences. ML bootstrap and BI posterior
2006). Molecular data of other genera classified in
Boliniales based on morphology (i.e. Apiocamarops
Samuels & J.D. Rogers, Mollicamarops Lar.N. Vassil-
jeva and Neohypodiscus J.D. Rogers, Y.M. Ju & Læssøe)
are not available. Within Boliniales, members of the
genus Endoxyla form a strongly supported group that
is not closely related to the morphologically similar
species A. curreyi. Apiorhynchostoma curreyi, the type
species of the genus, and P. ferruginea group with
Corn. ellipsoides, a species with soft-textured stromata
and hyaline, unicellular ascospores (Huhndorf et al.
2005). Camarops, which was inferred to be paraphy-
letic by Huhndorf and Miller (2008), is represented in
our phylogeny by seven species divided between two
clades. One of these clades encompasses two species
of Camaropella. Analyses of the combined ITS and
LSU sequences confirms the close relationship of
species of Endoxyla (FIG. 2) and resolves these species
as close relatives of Camarops polysperma and C.
ustulinoides. These species comprise a strongly sup-
ported clade that is sister to a lineage encompassing
A. curreyi and P. ferruginea.
Our results are consistent with investigations that
reported the absence of conidial anamorphs in Boli-
niales (Horn 1984, Petrini 1986, Samuels and Rogers
1987, Callan and Rogers 1989, Rogers et al. 1994a, Ju et
al. 1996, Del Valle Catania and Romero 2003). None of
taxa that we studied in vitro formed fertile ascomata.
Taxa within Boliniales are delimited primarily on
the basis of stromatal, ascomatal and ascospore
morphology, but these characters vary considerably
within some genera and it has yet to be determined
whether the separation of taxa using these features is
supportable. For example, the stromata of Camarops,
the largest genus in the order, range from applanate
to peltate or turbinate, can be hard and strongly
carbonized or soft-textured, and dark to lightly or
brightly colored. Species of Camaropella were segre-
gated from this genus because of their immersed,
valsoid stromata and clustered long-necked ascomata
(Vasilyeva 1997, Vasilyeva et al. 2007), but they are
inferred in this study and others (Huhndorf and
Miller 2008, Raja et al. 2011) to be close relatives of
species of Camarops not sharing these morphological
characteristics. The ascospores of species of Endoxyla,
the second-largest genus in Boliniales, are nonseptate,
one- or two-septate and may possess a hyaline cell. Our
phylogenies demonstrate that species of Endoxyla
possessing two-septate ascospores with a hyaline cell
are more closely related to E. operculata than to A.
curreyi, a species with morphologically similar asco-
spores, but a limited number of the representatives of
both genera were included in this study. Another
feature that warrants attention in future phylogenetic
studies of the Boliniales is the hyaline cap or
noncellular appendage that covers the germ pore of
the ascospores of some of the members of this order.
This structure, which is interpreted as a localized
thickening of the ascospore wall (Winter 1886; Mu ¨ller
and von Arx 1962; Rogers et al. 1994a, 2006; Rappaz
1995), is evident in all species of Apiorhynchostoma and
Neohypodiscus and also has been described for one
member of the genus Camarops. The hyaline cap is
often indistinct, but among the taxa examined in this
study, it is best seen in Melzer’s reagent or in
preparations stained with JGB.
Boliniaceae Rick emend. Unter. & Re ´blova ´
Stromata immersed to superficial, often erumpent
through the wood or bark of the host plant, crustose,
applanate, peltate, pulvinate, turbinate or clavate,
occasionally irregular and grotesque, hard- or soft-
textured, dark or brightly colored. Ascomata immersed
within the stromata, perithecial, monostichous or
polystichous, with short or elongate necks. Asci
persistent, cylindrical to clavate, long-stipitate, with an
indistinct, inamyloid apical ring. Ascospores hyaline or
various shades of brown, ellipsoid to suballantoid,
sometimes compressed laterally, aseptate, one- or two-
septate, with an inconspicuous apical germ pore that
may be covered by an indistinct hyaline cap, occasion-
ally with a germ pore at both ends, uniseriate within the
asci or biseriate and becoming uniseriate below.
Paraphyses abundant, filiform, longer than the asci.
On the bark and decorticated wood of angiosperms
and gymnosperms, occasionally on polypores, world-
wide. Conidial anamorphs not observed in situ and in
KEY TO GENERA OF BOLINIACEAE
1.Stromata immersed, poorly developed to valsoid or
pustulate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Stromata erumpent or superficial, crustose to
turbinate or clavate or irregular and grotesque
2.Stromata poorly developed, typically evident
only as a darkening of the wood surface or the
wood surrounding the ascomata . . . . . . . . .
2.Stromata well developed, valsoid or pustu-
late . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Ascospores brown, two-septate, with a hyaline cell,
porate end covered by an indistinct hyaline
cap . . . . . . . . . . . . . . . . . . . . . . . Apiorhynchostoma
Ascospores brown, aseptate, one- or two-septate,
with or without a hyaline cell, porate end not
covered by a hyaline cap . . . . . . . . . . . . . Endoxyla
4. Ascospores brown, aseptate . . . . .
4. Ascospores brown, one-septate
Ascospores hyaline . . . . . . . . . . . . . . Cornipulvina
. . 5
. . . Pseudovalsaria
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
5. Ascospores brown or consisting of a brown cell and
a smaller hyaline cell. . . . . . . . . . . . . . . . . . . . . 6
6. Ascospores one-septate, consisting of a larger,
porate brown cell and a smaller hyaline
cell . . . . . . . . . . . . . . . . . . . . . .
6. Ascospores aseptate, one- or two-septate,
entirely brown . . . . . . . . . . . . . . . . . . . . . .
Stromata hard, ascospores with a hyaline cap at
one or both ends . . . . . . . . . . . . . .
Stromata hard or soft-textured, ascospores lacking
a hyaline cap. . . . . . . . . . . . . . . . . . . . . . . . . . . 8
8. Stromata thin, soft-textured and brightly
les . . . . . . . . . . . . . . . . . . . . . . Mollicamarops
8.Stromata thick or thin, typically hard, occa-
sionally soft-textured, ostioles non-stella-
te . . . . . . . . . . . . . . . . . . . . . . . . . . . Camarops
with stellate ostio-
Apiocamarops Samuels & J.D. Rogers, Mycotaxon
Type species: Apiocamarops alba Samuels & J.D. Rogers
Specimens examined: Apiocamarops alba. BRAZIL. Amazo-
nas: Plateau of Serra Araca. 19–24 Feb 1984, G.J. Samuels
486, G.T. Francis and J. Pipoly (NY, HOLOTYPE); data as
for holotype, G.J. Samuels 438, G.T. Francis and J. Pipoly
(NY). FRENCH GUIANA. Circa 17 km SW of Sa ¨ul toward
Mount Galbao, 24, 26, 28 Jan 1986, G.J. Samuels 3301 and
J.R. Boise (NY); On road between Sinnary and St Elie,
ECEREX OSTROM Research Area, 20–29 Feb, 1 Mar 1986,
G.J. Samuels 3816 (NY); ca. 10 km SW of Sa ¨ul towardMount
Galbao, Jan–Feb 1986, G.J. Samuels 3696 and J.R. Boise (NY);
Paul Isnard Area, Mounts Lucifer and Decou Decou, ca.
150 km S of St Laurent du Maroni, 7–17 Mar 1986, G.J.
Samuels 4114 (NY); vic. Cayenne, 25 Mar 1986, G.J. Samuels
4454 and C. Feuillet (NY). Apiocamarops cryptocellula.
GUYANA. Cuyuni-Mazaruni region, foothills S of Mount
Ayanganna, ca. 1 km W of the Pong River, 26 Feb 1987, G.J.
Samuels 4836, J. Pipoly and G. Gharbarran (NY, HOLOTYPE).
Comments: Apiocamarops includes species charac-
terized by their superficial, soft-textured to woody,
white to yellowish or orange stromata, and ascospores
that consist of a larger, brown porate cell and a
smaller, hyaline nonporate cell (Samuels and Rogers
1987, Rogers and Samuels 1988, Rogers and Ju 2003).
Apiocamarops alba, the only species studied in axenic
culture, does not produce a conidial anamorph
(Samuels and Rogers 1987). The two other members
of the genus, A. cryptocellula J.D. Roger & Samuels and
A. pulvinata J.D. Rogers & Ju, have not been cultured.
Species of Apiocamarops occur on bark and
decorticated wood, and are known from Central and
South America (i.e. Brazil, Costa Rica, French Guiana,
Guyana, Venezuela) (Samuels and Rogers 1987,
Rogers and Samuels 1988, Rogers and Ju 2003). The
members of this genus are similar anatomically to
Camarops (Samuels and Rogers 1987), but the
relationship among them has yet to be evaluated
based on molecular data.
KEY TO SPECIES OF APIOCAMAROPS
1.Ascomata superficial, seated directly on wood and
united in a white to yellowish furfuraceous stroma,
ascospores (5–)6–9(–11.5) 3 (2.7–)3–3.7(–4)
mm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Ascomata immersed in a woody, pulvinate to
discoid stroma . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Ascospores 4.5(–5) 3 2.5(–3) mm . . . .
2. Ascospores 6.6–8 3 3.7 mm, hyaline cell often
collapsing . . . . . . . . . . . . . . . . . A. cryptocellula
Apiorhynchostoma Petrak, Ann. Mycol. 21:185. 1923.
Type species: Apiorhynchostoma curreyi (Rabenh.) E.
Comments: Species of Apiorhynchostoma possess
immersed, solitary to gregarious, flask-shaped asco-
mata surrounded by a reduced hyphal stroma that is
evident as a darkening of the substrate around the
ostioles and between the ascomata, and two-septate,
uniporate ascospores that consist of two brown cells
and a smaller, terminal hyaline cell (Mu ¨ller and von
Arx 1962, Sivanesan 1975, Rappaz 1995). The
members of this genus are distinguished from
Endoxyla in possessing ascospores with an apical pore
covered by an indistinct hyaline cap. Also, the species
examined are characterized by asci with an apical ring
that stains in JGB. Species of Apiorhynchostoma occur
on the decayed, decorticated wood of gymnosperms
in Europe and North America.
Apiorhynchostoma has been included in the Amphi-
sphaeriaceae and the Clypeosphaeriaceae (Xylariales)
(Barr 1990, 1994; Untereiner 1993; Rogers et al.
1994b; Hyde et al. 1998; Re ´blova ´ 1998; Kang et al.
1999; Wang et al. 2004) but our phylogenies position
A. curreyi in the Boliniales. Within this order A.
curreyi is most closely related to Cornipulvina and
Pseudovalsaria (FIGS. 1, 2).
KEY TO SPECIES OF APIORHYNCHOSTOMA
Ascospores (13–)15–16(–19) 3 4–6 mm. . . . . A. altipetum
Ascospores larger . . . . . . . . . . . . . . . . . . . . . . . . 2
2.Ascospores 20–24(–27) 3 7–10 mm . . . A. curreyi
2.Ascospores 25–35 3 9–12 mm . . . . A. tumulatum
Apiorhynchostoma altipetum (Peck) Rappaz, Mycologia
Helvetica 7:153. 1995.
emerging from the decorticated wood. 5–7. Ascospores, arrows indicate the hyaline cap. 8. Upper part of the ascus bearing
spores. 9. Ascus with distinct apical annulus. 10–13. Endoxyla macrostoma. 10, 11. Tips of ascomatal necks emerging from the
decorticated wood. The ascomata are monostichous, immersed and arranged in rows. 12. Longitudinal section of ascomata.
13. Ascospores. 3–9 from S.-A˚. Hanson 2007-278; 10–13. from M. Re ´blova ´ 1066. Bars: 3, 4, 10–12 5 500 mm; 5–9, 13 5 10 mm.
Apiorhynchostoma curreyi and Endoxyla macrostoma. 3–9. Apiorhynchostoma curreyi. 3, 4. Tips of ascomatal necks
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
;Sphaeria altipeta Peck, Bot. Gaz. 5:34. 1880.
;Rhynchostoma altipetum (Peck) Sacc., Syll. Fung. 1:731.
; Entosordaria altipeta (Peck) Ho ¨hnel, Sitz. Kaiserl. Akad.
Wiss. Math.-Naturwiss. Cl. Abt 1, 129:183. 1920.
5 Apiorhynchostoma trabicola You Z. Wang, Aptroot & K.D.
Hyde., Fungal Divers. Res. Ser. 13:142. 2004.
See Rappaz (1995) and Wang et al. (2004) for
descriptions and illustrations of this species.
Habitat: On rotten, decorticated wood of conifers.
Distribution: Czech Republic, Sweden, Switzerland,
USA (type locality).
Specimens examined: SWEDEN. E.M. Fries, Scleromyceti
Sueciae 268 (1st ed.), (FH, as Sphaeria operculata, collection
mixed). SWITZERLAND. Zu ¨rich: Tagelswangen, on wood
of Abies alba, 5 Dec 1955, A. Tonolo (Z, as Apiorhynchostoma
apiculata). USA. New Hampshire: Mount Washington, on
decaying wood, C.G. Pringle 328 (NYS, HOLOTYPE of
Comments: Johnova ´ (2009) reported this species
from the Czech Republic but did not cite collection
data. Apiorhynchostoma altipetum is distinguished
from the other members of the genus by its smaller
ascospores. The configuration of the ascomata of A.
altipetum is reminiscent of Endoxyla operculata, and
we examined one collection (Scl. Sueciae 268, 1st ed.)
that contained a mixture of wood fragments bearing
the ascomata of both species. Because we cannot
separate A. trabicola from A. altipetum based on the
description provided by Wang et al. (2004), we
consider these taxa to be conspecific.
Apiorhynchostoma curreyi (Rabenh.) E. Mu ¨ller, Beitr.
Kryptogamenfl. Schweiz 11:707. 1962.
; Sphaeria curreyi Rabenh., Fung. Europ. 250. 1860.
1859, non Wallroth, Fl. Cryptogamica Germ. 2:778. 1833.
; Anthostoma apiculata (Curr.) Niessl apud Kunze, Fungi
Sel. exs. 267. 1880 (fide Mu ¨ller and von Arx 1962).
;Valsaria apiculata (Curr.) Sacc., Syll. Fung. 1:752. 1882.
; Rhynchostoma apiculatum (Curr.) Winter, Rabenh.
Krypt.-Fl. 1:762. 1887.
; Xylosphaeria apiculata (Curr.) Cooke, Grevillea 17:86.
; Phaeosperma apiculatum (Curr.) Sacc. & Traverso, Syll.
Fung. 20:326. 1911.
; Entosordaria apiculata (Curr.) Ho ¨hnel, Sitz. Kaiserl.
Akad. Wiss. Math.-Naturwiss. Cl. Abt 1, 129:166. 1920.
; Apiorhynchostoma apiculatum (Curr.) Petrak, Ann.
Mycol. 21:185. 1923.
; Lopadostoma apiculatum (Curr.) P. Martin, J. S. African
Bot. 42:75. 1976.
5 Anthostoma trabeum Niessl, Beitr. z. Kenntn. Pilze p. 56.
See Mu ¨ller and von Arx (1962), Dennis (1981) and
Hyde et al. (1998) for descriptions of the tele-
omorph. Additional illustrations of this species are
provided by Currey (1859), Niessl (1872), Winter
(1886), Mu ¨ller and von Arx (1962), Dennis (1981),
Rogers et al. (1994b) and Hyde et al. (1998).
Colonies on CBSOA 6 mm in 21 d, flat, appearing
waxy, mycelium immersed entirely, orange-white
(5A2). Medium faintly cleared to 5 mm from the
growing edge of colonies. Colonies on CMA 32–
34 mm in 21 d, flat, appearing moist, mycelium
immersed entirely, light orange (5A4–5) at the center,
mycelium toward the margin brownish gray to brown-
orange (5B-C3-4). Colonies onMEA39–43 mm in 21 d,
aerial mycelium short, cottony, felt-like, restricted to
the colony center, white (A1), mycelium toward the
margin immersed, orange-white (5A2). Reverse
orange-white to light orange (5A2-4). Colonies on
MLA 20–21 mm in 21 d, aerial mycelium sparse, short,
restricted to the colony center, appearing felt-like,
light orange to melon (5A4–6), mycelium toward the
margin immersed, orange-white to orange gray (5A–
B2). Reverse brownish orange to grayish or light brown
(5B–C3–5). Colonies on OA 11–15 mm in 21 d, aerial
mycelium sparse, short, restricted to the colony center,
appearing powdery, white (A1), mycelium toward the
margin immersed, appearing waxy, brownish orange
(5C4–6). Reverse brownish orange to grayish or light
brown (5C–D3–4). Margin sharp and regular on MEA,
irregular and not sharply defined on all other media.
Conidial anamorph not observed.
Habitat: On rotting, decorticated wood of conifers.
Distribution: Austria, Canada, Germany, Sweden,
UK (type locality).
Specimens examined: AUSTRIA. Styria: Voitsberg, in
beams of fir (Abies sp.), summer, G. de Niessl, Rabenhorst
Fungi Europaei 1531 (DAOM, F, as Anthostoma trabeum).
CANADA. British Columbia: Haida Gwaii, Graham Island,
on an exposed log over a stream, along the Yakoun River,
about 5 m north of Yakoun Lake, 16 Jul 1967, I.M. Brodo
11643 and M.J. Shchepanek (DAOM 133701). GERMANY.
Hamburg: Sachsenwald, near Hamburg, on wood of Picea
excelsa (5 P. abies), O. Jaap, Rehm Ascomyceten 1614b (FH,
as Rhynchostoma apiculatum); Oberbayern: Thal near
Thalham, on decaying conifer wood, Jun 1905, H. Rehm,
Rehm Ascomyceten 1614 (FH, as R. apiculatum); Saxony:
Ko ¨nigstein, on old barriers made of conifer wood, May and
Jun 1885 and 1886, W. Krieger, Fungi Saxonici 176
Anthostoma apiculatum (FH, TRTC ex herb. R.F. Cain
No. 4745); Saxony: Helfta near Eisleben, on decorticated
wood of Pinus sylvestris, Oct 1879, J. Kunze, Fungi Selecti
Exsiccatii 267 (DAOM, FH, as A. apiculatum. SWEDEN,
Scania: A˚hus, Gropaha ˚lets NR, on wood of Pinus sylvestris,
25 Jul 2007, S.-A˚. Hanson 2007-278 (C; living culture
UAMH 11088 [mass-ascospore isolate]). UK. Batheaston, on
a decaying wooden fence, 1866 and 1857, F. Currey,
Rabenhorst Fungi Europaei 250 (DAOM, ISOTYPE, as
Sphaeria curreyi); Surrey: Weybridge, Oct 1857, F. Currey (K,
HOLOTYPE of S. apiculata); Surrey: Shere, May 1865, M.C.
Cooke, Fungi Britannici Exsiccati 272 (K, as S. apiculata);
Norfolk: King’s Lynn, C.B. Plowright, Sphaeriacei Britannici
80 (K, as S. apiculata).
Comments: Judging from the availability of specimens
and the number of descriptions of this species in the
literature, A. curreyi is the most frequently collected
memberofthe genus.Althoughthehyaline capcovering
the porate end of the ascospores of this species (FIGS. 5–
7) has not been illustrated previously, this structure is
evident in all collections examined in this study. Earlier
descriptions of the ascospores of this species (Winter
1886, Mu ¨ller and von Arx 1962) indicate that the cap was
interpreted as a localized thickening of the ascospore
wall. Niessl (1872) described the ascospores of this
species as possessing a gelatinous envelope, but we have
never observed this feature in A. curreyi.
As noted by Muller and von Arx (1962), Apior-
hynchostoma is based on Sphaeria apiculata, a later
homonym of S. apiculata Wallroth (5 Plagiostoma
salicellum [Fr.] Sogonov). The synonymy of the
former species with Sphaeria curreyi was recognized
first by Winter (1886). Anthostoma apiculatum, as
represented by Fungi Sel. exs. 267, is cited as a
synonym of A. curreyi (Winter 1886, Muller and von
Arx 1962), but examples of this exsiccata examined in
the present study (DAOM, FH) are conspecific with a
specimen in Schweiniz’s herbarium (PH) under the
name Sphaeria lineata (Alb. & Schwein.) Cooke.
Sphaeria lineata possesses immersed, nonstromatic
ascomata, cylindrical asci with an inamyloid apical
ring, and large (15–23 3 7–9 mm), brown, unicellular,
ellipsoid ascospores with a terminal germ pore, and
was identified by Untereiner (1993) as a species of
Anthostoma Nitschke. Rogers et al. (1994b) also
examined A. apiculatum (NY) but did not comment
on its identity, raising the possibility that Fungi Sel.
exs. 267 represents more than a single taxon.
Apiorhynchostoma tumulatum (Cooke) Sivan., Trans.
Brit. Mycol. Soc. 65:26. 1975.
; Sphaeria tumulata Cooke, Grevillea 7:4. 1878.
; Amphisphaeria tumulata (Cooke) Sacc., Syll. Fung.
5 Anthostomella brachystoma Ellis. & Everh., Bull. Washburn
Coll. Lab. Nat. History 1:5. 1884 (fide Rogers et al.
;Xylosphaeria brachystoma (Ellis & Everh.) Cooke,
Grevillea 17:86. 1889.
See Sivanesan (1975) and Rogers et al. (1994b) for
descriptions and illustrations of this species.
Habitat: On rotting, decorticated wood of Pinus
contorta Dougl. and Tsuga mertensiana (Bong.)
Comments: Apiorhynchostoma tumulatum is distin-
guished from the other members of this genus by its
large ascospores. To date, this species is known only
from North America. The hyaline cap covering the
porate end of the ascospores of A. tumulatum noted
by Harkness and Cooke (1878), Ellis and Everhart
(1892) and Sivanesan (1975) is not evident in the
illustrations of this species provided by Rogers et al.
(1994b), suggesting that this structure may be
difficult to observe in some collections.
Camaropella Lar.N. Vassiljeva, Mikologiya i Fitopato-
logiya, 31:6. 1997.
Type species: Camaropella pugillus (Schwein.: Fr.)
Comments: Camaropella encompasses species with
immersed, valsoid stromata, clustered, polystichous
ascomata with long necks that converge in an
erumpent stromatic cushion, and brown, nonseptate,
uniporate ascospores (Vasilyeva 1997, Vasilyeva et al.
2007). The two species assigned to the genus,
Camaropella lutea (Alb. & Schwein.) Lar.N. Vassiljeva
and Camaropella pugillus, are distinguished by the
color of the stromata and the degree to which the
ostioles project above the wood surface (Vasilyeva et
al. 2007). Both species have been studied in pure
culture; neither produces a conidial anamorph, but
the former forms fertile ascomata (Petrini 1986, Del
Valle Catania and Romero 2003). Camarops rostratus
Romero & Samuels, a species with immersed, pulvi-
nate stromata and long-necked ascomata (Romero
and Samuels 1991), also might belong to Camaropella,
according to Vasilyeva et al. (2007). Species of
Camaropella are known from rotting wood in Europe,
North America and South America (Shear 1940,
Nannfeldt 1972, Vasilyeva 1997, Del Valle Catania
and Romero 2003, Vasilyeva et al. 2007, Huhndorf
and Miller 2008, Johnova ´ 2009).
Huhndorf and Miller (2008) confirmed the place-
ment of Camaropella in Boliniales based on the
analysis of LSU sequences. They also identified C.
amorpha, C. microspora and Corn. ellipsoides as the
closest relatives of Camaropella lutea and Camaropella
pugillus, but the clade comprising these species was
weakly supported (Huhndorf and Miller 2008). The
results of our analyses of LSU sequences of 77
members of the Sordariomycetes including 16 repre-
sentatives of the Boliniales (FIG. 1) confirm the
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
position of C. amorpha and C. microspora as close
relatives of Camaropella and indicate that Camaropella
could be expanded to include these two species of
Camarops. For a key to the species of Camaropella, see
Vasilyeva et al. (2007).
Camarops P. Karst., Bd. Ka ¨nn. Finl. Nat. Folk 23:6.
Type species: Camarops hypoxyloides P. Karst. (5 C.
polysperma [Mont.] J.H. Mill.)
Comments: Camarops, which currently includes
more than 25 species, encompasses taxa characterized
by their superficial, applanate or pulvinate to peltate
or turbinate stromata that typically consists of a
compact (and sometimes strongly carbonized) ectos-
troma and a loose entostroma, obpyriform to tubular
ascomata, and small, brown, uni- or biporate asco-
spores that are usually noticeably flattened (Shear
1938, Nannfeldt 1972, Hilber and Hilber 1980,
Rogers and Samuels 1987, Samuels and Rogers
1987, Rogers and Ju 2003, Rogers et al. 2006). Within
the genus, C. flava Samuels & J.D. Rogers and C.
rogersii are distinctive in possessing thin, soft and
brightly colored stromata (Samuels and Rogers 1987,
Huhndorf and Miller 2008). The former species is
also the only member of the genus with ornamented
ascospores (Samuels and Rogers 1987). Camarops
antillana J.D. Rogers, Lechat & J. Fourn. forms soft-
textured, pulvinate, externally blackened stromata
and ascospores with a germ pore that is covered by a
hyaline thickening of the perispore (Rogers et al.
2006). As noted above, two members of the genus
with immersed, valsoid stromata and clustered, long-
necked ascomata have been segregated to Camar-
opella (Vasilyeva 1997, Vasilyeva et al. 2007).
A number of Camarops have been studied in pure
culture; none have been observed to form a conidial
anamorph, but two species, C. petersii and C.
spathulata (Berk. & Broome) Nannf., produce tele-
omorphs in vitro (Horn 1984, Callan and Rogers
1989, Del Valle Catania and Romero 2003). Del Valle
Catina and Romero (2003) provided a key to the
members of the genus from Argentina.
Species of Camarops are found worldwide and have
been collected in Africa, Asia, Europe, North America
and South America, where they occur on the
corticated branches and trunks of angiosperms, on
decaying, decorticated wood of angiosperms and
gymnosperms (Shear 1938; Nannfeldt 1972; Rogers
1981; Rogers and Samuels 1987; Samuels and
Rogers 1987; Del Valle Catania and Romero 2003;
Rogers et al. 2006, 2007) and occasionally on
polypores (Samuels and Rogers 1987, Lodge et al.
2002). A number of European species with conspic-
uous stromata are restricted to old growth and
minimally managed stands, and their rarity has been
attributed to modern silvicultural practices and the
removal of woody debris and fallen, decaying tree
from forests (Nannfeldt 1972, Holec 2005, Johnova ´
2009). In Germany, the occurrence of C. polysperma
in near-natural forests is related to the availability of
large-diameter logs (Ba ¨ssler et al. 2010).
Phylogenetic analysis of the LSU sequences of the
taxa examined in this study divides species of Camarops
between two moderately well supported clades. The
first includes C. amorpha, C. microspora and species of
Camaropella. The second includes C. petersii, C.
polysperma, C. rogersii, C. tubulina and C. ustulinoides.
Within this second group, two isolates of C. ustulinoides
are not resolved as close relatives. Although Camar-
opella could be expanded to include C. amorpha and C.
microspora, wehesitatetoformally transfer thesespecies
regions of the members of both genera.
Cornipulvina Huhndorf, A.N. Mill., F.A. Ferna ´ndez &
Lodge, Fungal Divers 20:61. 2005.
Type species: Cornipulvina ellipsoides Huhndorf, A.N.
Mill., F.A. Ferna ´ndez & Lodge
Comments: This monotypic genus is characterized
by its superficial, soft-textured stromata, monosti-
chous ascomata with long, projecting necks, and
hyaline, unicellular ascospores (Huhndorf et al. 2005,
Huhndorf and Miller 2008). Cornipulvina ellipsoides
occurs on decorticated wood and has been collected
in Brazil, Puerto Rico and Venezuela (Huhndorf et al.
Cornipulvina ellipsoides was resolved as the member
of a weakly supported clade that also encompassed C.
amorpha, C. microspora and species of Camaropella in
a study based on the analysis of LSU sequences
(Huhndorf and Miller 2008). Our LSU phylogeny
positions this species as the member of a clade that
includes A. curreyi and P. ferruginea (FIG. 1).
Endoxyla Fuckel, Jb. Nassau. Ver. Naturk. 25–26:321.
Type species: Endoxyla macrostoma Fuckel
Comments: Endoxyla encompasses species occur-
ring on decayed, decorticated wood of angiosperms
and gymnosperms in Europe and North America
that possess immersed, poorly developed to pulvinate
stromata, solitary to gregarious, flask-shaped asco-
mata, and uniporate, aseptate, or one- or two-septate
ascospores that may or may not possess a hyaline cell
(Untereiner 1993). Members of the genus studied in
pure culture (see below) have not been observed to
produce conidial anamorphs. Species of Endoxyla are
ecologically and morphologically similar to Apior-
hynchostoma (Dennis 1981, Rogers et al. 1994b) but
differ in possessing ascospores that lack a hyaline cap
and asci with an apical ring that does not stain in
Endoxyla has been treated as the member of a
number of ascomycete families (Barr 1990, 1994:
Untereiner 1993; Rogers et al. 1994b). Our phyloge-
nies (FIGS. 1, 2) support Rappaz (1995) who referred
this genus to the Boliniaceae based on morphology.
KEY TO SPECIES OF ENDOXYLA (ADAPTED
FROM UNTEREINER 1993)
1. Ascospores aseptate, (8–)10–12.5(–14.5) 3 2–3.7
(–5) mm . . . . . . . . . . . . . . . . . . . . . . E. operculata
Ascospores septate, septum delimiting a hyaline
cell or dividing a pigmented portion of the
ascospore, or both . . . . . . . . . . . . . . . . . . . . . . . 2
2. Ascospores without a hyaline cell, one-septate,
(9.5–)11–14.5(–15.5) 3 2.3–3.5(–4.5) mm . . . . .
. . . . . . . . . . . . . . . . . . . . . . E. parallela
2.Ascospores with a hyaline cell, one- or two-
septate . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Pigmented portion of the ascospore septate . . . . . 4
Pigmented portion of the ascospore aseptate . . . . 5
4. Ascospores (10–)11.3–14.8(–17.8) 3 2.6–3.6
(–4) mm . . . . . . . . . . . . . . . . . . . . . E. munkii
4.Ascospores (22–)24–31 3 4.5–5(–7) . . . E. occulta
Ascospores ellipsoid or ellipsoid-inequilateral to
crescentic, (15–)20–25(–36.5) 3 7.5–9(–10.5)
mm . . . . . . . . . . . . . . . . . . . . . . . . . . . E. mallochii
Ascospores ellipsoid-cylindric or suballantoid,
smaller . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
6.Ascospores suballantoid, 9.0–12.3(–13.3) 3
2.2–3.4(–4.0) mm . . . . . . . . . . . . E. macrostoma
6.Ascospores ellipsoid-cylindric, 8.3–11 3 2.3–
3.5 mm . . . . . . . . . . . . . . . . . . E. xanthostroma
Endoxyla macrostoma Fuckel, Jb. Nassau. Ver. Naturk.
; Valsa macrostoma (Fuckel) Winter, Rabenh. Krypt.-Fl.
1(2): 689. 1886.
See Untereiner (1993) for a description of the
teleomorph. Additional illustrations of this species
are provided by Berlese (1905) and Untereiner
Colonies on CBSOA 5–6 mm in 21 d, flat,
appearing moist and waxy, mycelium immersed
entirely, yellowish white to pale yellow (3A2–3).
Colonies on CMA 7–8 mm in 21 d, flat, appearing
moist and waxy, mycelium immersed entirely, yellow
(3A6–8). Reverse light yellow (3A5). Colonies on OA
6–8 mm in 21 d, aerial mycelium sparse, short,
appearing powdery, yellowish white (3A2), mycelium
at the colony margin yellow (3A6–8). Reverse pale
yellow (3A3). Colonies on MEA 25–27 mm in 21 d,
mycelium immersed entirely, flat, center olive brown
(4F4–5), margin yellowish gray (4B2). Colonies on
MLA 20–25 mm in 21 d, aerial mycelium short,
sparse, restricted to the colony center, yellowish white
to pale yellow (3A2–3), mycelium toward the margin
immersed, slightly furrowed, appearing mealy,
orange-white (5A2) but becoming pale orange to
light orange (5–6A3–4) toward the colony margin.
Margin sharp and regular on all media except MLA.
Conidial anamorph not observed.
Habitat: On decorticated wood of angiosperms and
Distribution: Canada, Czech Republic, Germany
(type locality), Sweden.
Specimens examined: CANADA. Prince Edward Island:
Queens County, Brookvale Demonstration Woodlot, on
decorticated wood of Pinus strobus, 28 Apr 2009, A. Carter
(NBM; living culture UAMH 11491 [mass-ascospore iso-
late]); Queens County, Black Creek Road, on decaying
gymnosperm wood (Abies sp. or Picea sp.), 11 May 2008, A.
Carter (NBM). CZECH REPUBLIC. Southern Bohemia:
Sˇumava Mts., Zˇelezna ´ Ruda, glacial cirque of the Cˇerne ´
Jezero Lake, 19 Jul 1997, on decaying wood of Picea abies,
M. Re ´blova ´ 1066. See Untereiner (1993) for additional
Endoxyla mallochii Unter., A. Carter & Re ´blova ´, sp.
Stromata absent or poorly developed and evident
only as a light brown discoloration of the wood
around the necks of the ascomata. Ascomata im-
mersed, rarely solitary, monostichous in regular or
irregular linear groups, ampulliform, 900–1100 3
600–650 mm, black, with cylindrical, separately erum-
pent necks 350–400 3 200–250 mm, apices of necks
pulvinate, projecting slightly above the surface of the
wood. Ascomatal walls 30–60 mm wide, composed of
two layers, the outer layer of brown, thick-walled cells
forming a textura angularis to a textura prismatica,
the inner layer of thin-walled, compressed hyaline
cells forming a textura prismatica. Asci unitunicate,
eight-spored, cylindrical, long-stipitate, total length
245–305(–375) mm, pars sporifera 130–190(–245) 3
9–12.5 mm, with an indistinct, inamyloid apical ring
that is unstained in JGB. Ascospores ellipsoid to
ellipsoid-inequilateral or crescentic, (15–)20–25(–
36.5) 3 7.5–9(–10.5) mm, uniseriate or occasionally
apically biseriate and becoming uniseriate below,
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
Ascospores from eight-spored asci. From A. Carter 880, NBM (Holotype). Bars: A 5 20 mm, B 5 500 mm, D 5 10 mm.
Endoxyla mallochii. A. Asci. B. Habit sketch of ascomata. C. Enlarged ascospores from a two-spored ascus. D.
Apical annulus. 18–20. Ascospores. 21, 22. Long-stipitate asci. 23. Paraphyses. 15, 23 from M. Re ´blova ´ 1392; 16, 22 from M.
Re ´blova ´ 1388; 17–21 from M. Re ´blova ´ 1466. Bars: 15, 16 5 500 mm; 17–20 5 10 mm; 21–23 5 25 mm.
Endoxyla mallochii. 15, 16. Groups of ascomata immersed in decorticated wood with only necks emerging. 17.
UNTEREINER ET AL.: MOLECULAR PHYLOGENY OF BOLINIALES
bicellular, composed of a hyaline cell 1.5–2.5 3 2–
2.5 mm, and a brown cell with an apical germ pore,
porate end acute, without a fixed orientation within
the asci. Paraphyses filiform, septate, 1.5–3 mm diam.
Colonies on CBSOA, MEA, MLA and OA orange-
white (5A2). Colonies on CMA white (A1). Colonies
flat and immersed entirely on all media except MLA.
Colonies on CBSOA and OA 5–6 mm in 21 d,
appearing waxy. Colonies on CMA 5–6 mm in 21 d,
appearing moist. Colonies on MEA 41–45 mm in 21 d.
Colonies on MLA 13–14 mm in 21 d, aerial mycelium
short, restricted to the colony center, appearing felt-
like, mycelium toward the margin immersed, appear-
ing waxy. Margin regular and sharply defined on all
media. Conidial anamorph not observed.
Etymology: Named in honor of David Malloch, a
distinguished Canadian mycologist and mentor to AC
Habitat: On decorticated wood of conifers.
Distribution: Canada (type locality), Czech Repub-
HOLOTYPE. CANADA. Prince Edward Island: Queens
County, Fort Amherst Park, on decorticated wood (Abies
sp. or Picea sp.), 2 Jun 2008, A. Carter 880 (NBM; living
culture UAMH 11087 ex-type, isolated from ascomatal
Additional specimens examined: CANADA. Prince Ed-
ward Island: Queens County, Fort Amherst Park, on
rotted fallen log (Pinaceae), 9 Jul 2008, A. Carter 914
(NBM); ibid., on decorticated wood (Abies sp. or Picea
sp.), 8 Jun 2008, A. Carter 951 (NBM); ibid., on
decorticated wood (Abies sp. or Picea sp.), 8 Aug 2008,
A. Carter 955 (NBM); Queens County, woods at Black
Creek Road, on fallen log (Picea sp.), 15 Oct 2009, A.
Carter 1086 (NBM); Queens County, Cornwall, Corwall
Ferry Road, on well rotted, decorticated wood (Pina-
ceae), 23 Apr 2009, A. Carter 1128 (NBM). CZECH
REPUBLIC. Northeastern Bohemia: Chr ˇibsko, Labske ´
Pı ´skovce Mountains, on decayed wood of Picea abies, 23
Apr 1998, M. Re ´blova ´ 1207. Southern Bohemia, Sˇumava
Mountains, Zˇelezna ´ Ruda, glacial cirque of the Cˇerne ´
jezero lake, on decaying wood of Picea abies, 27 Aug 1998,
M. Re ´blova ´ 1388, 1392; 13 Aug 1999, M. Re ´blova ´ 1466.
UKRAINE. Carpathian Mountains, Sinevirskaja Poliana
Massif., Sinevir, on decayed wood of Abies alba, 20 Jul
1998, M. Re ´blova ´ 1244.
Comments: Endoxyla mallochii resembles E. macro-
stoma and E. xanthostroma Unter. in possessing
uniseptate ascospores composed of a small hyaline cell
and a larger brown cell with an apical germ pore. This
species can be distinguished from the other members of
the genus by its larger, ellipsoid to crescentic ascospores
and longer, cylindrical asci. The asci in some collections
of E. mallochii occasionally contain only two or four
ascospores that are considerably larger than the asco-
spores in eight-spored asci. Abnormal ascospores of E.
mallochii may possess or lack a hyaline cell, but we have
never observed the pigmented portion to be septate.
Endoxyla munkii Unter., Mycologia 85:299. 1993.
This species is described and illustrated by Unte-
Habitat: On decaying, decorticated wood of species
Distribution: Finland, Russia (type locality), Swe-
Comments: Endoxyla munkii resembles E. parallela
(Fr.: Fr.) Sacc., the only other member of the genus
Endoxyla associated with Scotiosphaeria endoxylinae
Sivan. (Sivanesan 1977). Endoxyla munkii has been
compared to Apiorhynchostoma altipetum (Rappaz
1995), but it can be distinguished from this species
by its smaller and more centrally septate ascospores
that lack a hyaline cap.
Endoxyla occulta (Re ´blova ´) Unter. & Re ´blova ´, comb.
Basionym. Apiorhynchostoma occultum Re ´blova ´, Sydowia
See Re ´blova ´ (1998) for a description and additional
illustrations of the teleomorph.
Habitat: On decorticated, decaying wood of Picea
abies (L.) H. Karst.
Distribution: Czech Republic.
Specimens examined: CZECH REPUBLIC. Southern Bo-
hemia: Sˇumava Mountains, glacial cirque of the Cˇerne ´
jezero Lake, on decayed wood of a trunk of Picea abies, 22
Oct 1996, M. Re ´blova ´ (HOLOTYPE, PRM 842971); data as
for holotype, 27 Aug 1998, M. Re ´blova ´ 1387.
Comments: Endoxyla occulta resembles E. munkii
but it can be distinguished from this species by its
larger ascospores. Re ´blova ´ (1998) interpreted the
somewhat more refractive walls at the porate end of
the ascospores of this species as an appendage, but a
hyaline cap equivalent to the structure observed in
species of Apiorhynchostoma is not present. Endoxyla
occulta is known only from the type locality.
Endoxyla operculata (Fr.: Fr.) Sacc., Syll. Fung. 1:181.
;Sphaeria operculata Pers. dd seriata Alb. & Schwein.,
Conspect. Fung. Lusat., p. 40. 1805.
;Sphaeria operculata Fr.: Fr., Syst. Mycol. 2:479. 1823, non
Persoon, Syn. Meth. Fung. p. 80. 1801.
;Valsa operculata (Fr.: Fr.) Nitschke, Pyrenomyc. Germ.
;Anthostoma operculatum (Fr.: Fr.) J. Schro ¨t., Krypt.-Fl.
Schlesien 3:432. 1897.