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Phylogenetic classification and generic delineation of
Calyptosphaeria gen. nov., Lentomitella,Spadicoides and
Torrentispora (Sordariomycetes)
M. R
eblov
a
1*
, A.N. Miller
2
,K.R
eblov
a
3
, and V.
St
ep
anek
4
1
Institute of Botany of the Czech Academy of Sciences, Průhonice 252 43, Czech Republic;
2
Illinois Natural History Survey, University of Illinois, Champaign, IL 61820,
USA;
3
Central European Institute of Technology, Masaryk University, Brno 625 00, Czech Republic;
4
Institute of Microbiology of the Czech Academy of Sciences, Prague
142 20, Czech Republic
*Correspondence:M.R
eblov
a,
martina.reblova@ibot.cas.cz
Abstract: The genus Ceratostomella has a long history of taxonomic confusion. While species with evanescent asci have been transferred to the Microascales and
Ophiostomatales, the taxonomic status of species with persistent asci has not been completely resolved. In previous studies using DNA sequence data, cultures and
morphology, several Ceratostomella spp. were allocated in 13 genera in the Eurotiomycetes and Sordariomycetes. In our study, the systematics of the remaining
Ceratostomella spp. with persistent asci is revisited with new collection data, cultures and phylogeny based on novel DNA sequences from six nuclear loci. Bayesian
inference and Maximum Likelihood analyses support the monophyly of several wood-inhabiting species formerly classified in Ceratostomella and other unknown
morphologically similar taxa and their division into four genera, i.e. Lentomitella,Spadicoides,Torrentispora and the newly described Calyptosphaeria. This robust clade
represents the order Xenospadicoidales in the Sordariomycetidae. Comparative analysis of the ITS2 secondary structure revealed a genetic variation among Lentomitella
isolates; 11 species were recognised, of which five are newly introduced and two are new combinations. Other taxonomic novelties include four new species and eight
new combinations in Calyptosphaeria,Spadicoides, and Torrentispora. Molecular data suggest that Spadicoides is polyphyletic. The core of the genus is positioned in the
Xenospadicoidales;Spadicoides s. str. is experimentally linked with sexual morphs for the first time. Based on DNA sequence data, the monotypic genera
Xenospadicoides and Pseudodiplococcium are reduced to synonymy under Spadicoides, while Fusoidispora and Pseudoannulatascus are synonymised with
Torrentispora. Members of the Xenospadicoidales inhabit decaying wood in terrestrial and freshwater environments and share a few morphological characters such as
the absence of stromatic tissue, ascomata with a cylindrical or rostrate neck, similar anatomies of the ascomatal walls, thin-walled unitunicate asci with a non-amyloid
apical annulus, disintegrating paraphyses, usually ellipsoidal to fusiform ascospores and holoblastic-denticulate or tretic conidiogenesis. Revised Ceratostomella spp. with
persistent asci are listed and the taxonomic status of each species is re-evaluated based on revision of the holotype and other representative material, published details
and available phylogenetic data.
Key words: Ceratostomella, Conidiogenesis, Holoblastic-denticulate, Molecular systematics, New taxa, Phaeoisaria-like, Selenosporella-like, Tretic, Taxonomy,
Xenospadicoidales.
Taxonomic novelties: New genus: Calyptosphaeria R
eblov
a & A.N. Mill; New species: Calyptosphaeria collapsa R
eblov
a & A.N. Mill., C. tenebrosa R
eblov
a & A.N.
Mill., Lentomitella magna R
eblov
a, L. obscura R
eblov
a, L. striatella R
eblov
a, L. sulcata R
eblov
a, L. tenuirostris R
eblov
a, Torrentispora calembola R
eblov
a & A.N. Mill.,
T. novae-zelandiae R
eblov
a & A.N. Mill; New combinations: Calyptosphaeria subdenudata (Peck) R
eblov
a & A.N. Mill., C. tropica (Huhndorf et al.)R
eblov
a & A.N. Mill.,
Lentomitella conoidea (Feltg.) R
eblov
a, L. investita (Schw.) R
eblov
a, Spadicoides fuscolutea (Rehm) R
eblov
a, S. hyalostoma (Munk) R
eblov
a, Spadicoides iberica (Hern.-
Restr. et al.)R
eblov
a & A.N. Mill., Torrentispora aquatica (Vijaykr. et al.)R
eblov
a & A.N. Mill., T. biatriispora (K.D. Hyde) R
eblov
a & A.N. Mill., T. dubia (Sacc.) R
eblov
a&
A.N. Mill.
Available online 6 December 2017; https://doi.org/10.1016/j.simyco.2017.11.004.
INTRODUCTION
The perithecial ascomycete genus Ceratostomella (Saccardo
1878a) has a long history of taxonomic debate. Although the
simple generic diagnosis comprised only hyaline, aseptate as-
cospores, asci and perithecia, which are similar to those of
Ceratostoma (Fries 1818), Ceratostomella soon became a large,
heterogeneous assemblage of fungi for which Index Fungorum
lists 110 epithets. Although widely distributed throughout the
Northern Hemisphere, members of Ceratostomella are incon-
spicuous and difficult to find due to their small immersed to
superficial, long-necked ascomata. The asci are persistent or
evanescent containing septate or aseptate, hyaline or brown
ascospores, and most of the species are difficult to culture.
The homogeneity of Ceratostomella was soon challenged by
Kuntze (1898), who transferred 29 species with persistent asci
and mostly hyaline ascospores to Amphitrichum (Nees & Nees
1818). Amphitrichum was emended by Corda (1837) based on
A. olivaceum (= ? Cladosporium sp. fide Hughes 1958), but later it
was determined to be a nomen dubium fide Hughes (1958),asno
type specimen was given. Kuntze (1898) clearly misinterpreted
the generic concept of Amphitrichum, which is likely a dematia-
ceous hyphomycete. Another step towards clarification of the
concept of Ceratostomella was made by Höhnel (1906a).Len-
tomitella, originally described as a monotypic genus for Cera-
tostomella vestita, was introduced in order to segregate taxa with
ellipsoidal, 1-septate, hyaline, longitudinally striate ascospores
from species with similar ascospores containing more than one
septum and 2–4 large drops. Höhnel (1906a) suggested that
such taxa should belong to Ceratosphaeria and Lentomita.
However, von Arx (1952) did not accept Höhnel's narrow concept
and designated Ceratostomella as the correct generic name.
The broadly perceived Ceratostomella was redefined by
R
eblov
a (2006) based on the lectotype species, C. rostrata
Peer review under responsibility of Westerdijk Fungal Biodiversity Institute.
© 2017 Westerdijk Fungal Biodiversity Institute. Production and hosting by ELSEVIER B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
available online at www.studiesinmycology.org STUDIES IN MYCOLOGY 89: 1–62 (2018).
1
(Clements & Shear 1931), and three other accepted species.
Using comparative morphology, Ceratostomella was confined to
taxa with non-stromatic ascomata with a cylindrical neck, asco-
matal wall up to 100 μm thick, persistent clavate asci arising from
supporting ascogenous cells, broad-celled paraphyses and
brown, aseptate, ellipsoidal to reniform ascospores. Based on
DNA sequence data of two representative species, C. cuspidata
and C. pyrenaica, the genus was classified as Sordariomycetidae
incertae sedis. Species with evanescent asci and dark perithecia
with filiform necks were recombined and placed in Ceratocystis,
Huntiella and Thielaviopsis of the Microascales,orLep-
tographium,Ophiostoma and Pesotum of the Ophiostomatales
(Höhnel 1918, Elliott 1925, Moreau 1952, Hunt 1956, de Beer
et al. 2013a, b, 2014). Based on multigene phylogenetic ana-
lyses, the placement of the remaining Ceratostomella spp. with
persistent asci was partially resolved resulting in the recovery of
three robust phylogenetic lineages centred around the Amplis-
tromatales,Calosphaeriales and Ophiostomatales. The genus
Wallrothiella (Saccardo 1882)(Amplistromatales)basedon
W. congregata [= Ceratostomella sphaerosperma] was redefined
with the aid of DNA sequence data, recently collected material
and an acrodontium-like asexual morph (R
eblov
a & Seifert 2004,
Huhndorf et al. 2009). Several other Ceratostomella species were
reclassified in Jattaea and Togniniella (Calosphaeriales) and
Phaeoacremonium (Togniniales) based on the revision of type
material, evidence from molecular data and phialophora- or
acremonium-like asexual morphs producing phialidic conidia
in vitro (R
eblov
aet al. 2004, 2015a, R
eblov
a 2011, Gramaje et al.
2015). The “ophiostomataceous”lineage comprised Ceratosto-
mella s. str. and also Barbatosphaeria,Lentomitella,Natantiella,
and Xylomelasma (Höhnel 1906a, R
eblov
a 2006, 2007, Huhndorf
et al. 2008, Marincowitz et al. 2008, R
eblov
a&
St
ep
anek 2009).
The asexual morphs linked with genera of this lineage are
dematiaceous hyphomycetes with holoblastic conidia produced
on a sympodially extending rachis or on a terminal cluster of
denticles. They are part of the life cycle of Barbatosphaeria as
ramichloridium- and sporothrix-like (Samuels & Candoussau
1996, R
eblov
a 2007, R
eblov
aet al. 2015b) and Lentomitella as
phaeoisaria-like (R
eblov
a2006) asexual morphs. Other Cera-
tostomella spp. were dispersed to Ceratosphaeria (Magnapor-
thales)(Niessl 1876, Huhndorf et al. 2008), Chaetosphaeria
(Chaetosphaeriales)(Booth 1957, Huhndorf & Fern
andez 2005),
Daruvedia (Pyrenulales)(Dennis 1988) and Pseudorhynchia
(Hypocreales)(Samuels & Barr 1997).
The ongoing taxonomic revision of species of Ceratostomella
s. lat. with persistent asci revealed for many of them striking
morphological similarities with Lentomitella. Based on nucLSU
and nucSSU rDNA sequence data, comparative morphology and
cultures, Lentomitella was reinstated in the Sordariomycetidae
and shown to be distantly related to Ceratostomella (R
eblov
a
2006). The generic concept of Lentomitella was expanded to
include species with 1–3-septate, longitudinally striate, hyaline
ascospores, and also aseptate, smooth-walled, hyaline (R
eblov
a
2006) and brown ascospores (Huhndorf et al. 2008). Members of
Lentomitella bear a certain resemblance to Torrentispora (Hyde
et al. 2000) and Pseudoannulatascus (Luo et al. 2015) charac-
terised by ascomata with a long-neck, cylindrical asci and fusi-
form, hyaline, smooth- and thick-walled, usually aseptate
ascospores, rarely with delayed formation of septa (Hyde et al.
2000, Fryar & Hyde 2004, Barbosa et al. 2013). A monotypic
family, the Lentomitellaceae, was introduced by Zhang et al.
(2017).
In this study, several species historically treated in Ceratos-
tomella were recollected and isolated in axenic culture. Together
with other unknown, morphologically similar taxa resembling
Lentomitella and Torrentispora were subjected to phylogenetic
analyses. Fungi of this assemblage occur on decaying wood or
in bark in terrestrial habitats or on wood submerged in water.
They share a simple inconspicuous morphology of non-stromatic
ascomata with a cylindrical or rostrate neck, hyaline or brown,
aseptate or septate, ornamented or smooth-walled ascospores,
persistent asci with a non-amyloid apical annulus and partially
disintegrating paraphyses. Little is known about their asexual
morphs, which so far include only phaeoisaria-like morphs linked
with Lentomitella. However, the majority of these fungi are diffi-
cult to culture or they produce only sterile mycelium in vitro.
Recently, we found Ceratostomella fuscolutea (Rehm 1908),
Ceratostomella hyalostoma (Untereiner 1993), and an unknown
lentomitella-like species to produce Spadicoides asexual morphs
in vitro. This dematiaceous hyphomycete is characterised by
polytretic conidiogenous cells, unbranched conidiophores and
dark brown septate or aseptate conidia borne singly or in short
chains (Hughes 1958, Ellis 1963) and has not yet been linked
with any sexually reproducing ascomycetes as a part of their life
cycle. DNA sequence data suggest that Spadicoides is poly-
phyletic (Shenoy et al. 2010); S. atra was shown closely related
to Lentomitella.Hern
andez-Restrepo et al. (2017) confirmed the
placement of S. bina, the type species, in the Cordanales and
segregated S. atra from Spadicoides into a monotypic genus
Xenospadicoides in the Xenospadicoidales.
In order to unravel this part of the fungal tree of life, determine
the placement of Ceratostomella spp. listed above and other
similar taxa in monophyletic genera and resolve their familial and
ordinal relationships, we employed a polyphasic approach in this
study. We generated a multigene-based phylogeny of six nuclear
ribosomal and protein-coding loci of the new isolates and inten-
sively examined morphological characters of specimens and
isolates in pure culture. We also investigated intraspecificre-
lationships among members of Lentomitella using the Compen-
satory Base Change (CBC) criterion in the ITS2 secondary (2D)
structure in two most conserved helices II and III (Mai & Coleman
1997, Coleman 2009) and also in helix I (Müller et al. 2007). The
ITS2 is a fast-evolving part of the nuclear-coded rRNA operon,
which has proven useful for formulating molecular taxonomic
concepts, and its 2D structure has a potential to predict sexual
incompatibility among closely related organisms. The CBC hy-
pothesis is based on occurrence of compensatory base changes,
i.e. co-evolution of nucleotides involved in the double-sided
substitution in helices of the ITS2 molecule (Coleman 2000,
Müller et al. 2007). We performed in-depth comparative ana-
lyses of ITS2 2D structures of Lentomitella spp. and mapped all
existing substitutions among co-evolving nucleotides onto the
predicted 2D model of ITS2 of the type species L. vestita.
MATERIAL AND METHODS
Herbarium material and fungal strains
Herbarium material was rehydrated with water and examined
with an Olympus SZX12 dissecting microscope; hand-sectioned
ascomata, centrum material (including asci, ascospores and
paraphyses), conidiophores and conidia from living cultures were
R
EBLOV
AET AL.
2
mounted in 90 % lactic acid, Melzer's reagent or Lugol's iodine.
All measurements were made in Melzer's reagent.
Means ± standard deviation (SD) based on 20–25 measure-
ments are given for dimensions of asci, ascospores, con-
idiogenous cells and conidia. Microscopic structures were
examined using an Olympus BX51 compound microscope
(Olympus America, Inc., Melville, USA) with differential interfer-
ence contrast (DIC) and phase contrast (PC) illumination. Im-
ages of microscopic structures were captured with an Olympus
DP70 camera operated by Imaging Software Cell^D (Olympus).
Macroscopic images of colonies were documented using an
Olympus C-3030 digital camera with daylight spectrum 5600K
16W LED lights. All images were processed with Adobe Pho-
toshop CS6 (Adobe Systems, San Jose, USA).
Cultures were maintained on Modified Leonian's agar (MLA)
(Malloch 1981). For comparative purposes, strains were grown
on MLA and potato-carrot agar (PCA) (Gams et al. 1998). De-
scriptions of colonies are based on 28-d-old cultures. Ex-type
and other cultures are maintained at the Westerdijk Fungal
Biodiversity Institute (CBS), Utrecht, the Netherlands and the
International Collection of Microorganisms from Plants (ICMP),
Auckland, New Zealand. Type and other herbarium material are
deposited in the Herbarium of the Institute of Botany (PRA),
Průhonice, Czech Republic, the New Zealand Fungarium (PDD),
Auckland, New Zealand, and the Illinois Natural History Survey
Fungarium (ILLS), Champaign, Illinois, USA.
DNA extraction, amplification and sequencing
Total genomic DNA was extracted from either mycelium removed
from 14-d-old cultures grown on MLA or mature ascomata from
herbarium material using the UltraClean Microbial DNA Kit
(MoBio Laboratories Inc., Carlsbad, USA). For DNA extracted
from herbarium material, an alternative lysis method was incor-
porated: the gelatinous centrum of 10–15 ascomata was satu-
rated with distilled water, carefully removed with a needle and
placed in a 1.9 mL MicroBead tube provided by the manufac-
turer. After the fungal material was dissolved in 300 μLof
MicroBead Solution and 50 μL of Solution MD1, the preparations
were heated to 65 ºC for 10 min. The remaining steps for DNA
extraction from cultures and herbarium material followed the
manufacturer's protocol for filamentous fungi. All amplifications
were carried out in 0.5 mL thin-walled PCR tubes (Eppendorf
AG, Hamburg, Germany) using a PTC-200 thermal cycler (MJ
Research Inc., Watertown, USA). PCR reactions and primers
used for the amplification and sequencing of the internal tran-
scribed spacer (ITS) of the nuclear rRNA cistron, portions of the
nuclear ribosomal large subunit (nucLSU) and small subunit
(nucSSU) RNA gene, and segments 5–7 of the second largest
subunit of RNA polymerase II (rpb2) were carried out according
to the methods of R
eblov
aet al. (2017).
Primers used for the amplification and sequencing of other
genes included: 1) ACT-512F and ACT-783R (Carbone & Kohn
1999) for alpha-actin (act1) gene and 2) T1 and Bt2a in combi-
nation with Bt2b (Glass & Donaldson 1995, O’Donnell & Cigelnik
1997) for exons 2‒6 of beta-tubulin (tub2) gene. PCR reactions
containing 4 mM MgSO
4
were performed using Platinum
®
Taq
DNA polymerase High Fidelity (Invitrogen, Carlsbad, USA) in
25 μL volume reactions. PCR conditions were (act1) 2 min at
94 °C, 45–48 cycles of 30 s at 94 °C, 30 s at 54–55 °C and 30 s at
68 °C; (tub2) 2 min at 94 °C, 40–48 cycles of 30 s at 94 °C, 30 s at
54–56 °C and 45–60 s at 68 °C, with a final extension of 10 min at
68 °C for all amplifications. Amplicons were either purified directly
from PCR solution after amplification or isolated from agarose gel
using the High Pure PCR Product Purification Kit (Roche Applied
Science, Mannheim, Germany) following the manufacturer'sdi-
rections. Automated sequencing was carried out by GATC
Sequencing Service (Cologne, Germany). Raw sequence data
were assembled, examined and edited using Sequencher v. 5.4.1
software (Gene Codes Corp., Ann Arbor, USA).
GenBank accession numbers for act1, ITS, nucLSU,
nucSSU, rpb2 and tub2 sequences generated during this study
and homologous sequences of representatives of the Sordar-
iomycetes and Leotiomycetes retrieved from GenBank are listed
in Table 1. Retrievable sequences have been published in
various studies, e.g. Suh & Blackwell (1999), Huhndorf et al.
(2004), Miller & Huhndorf (2004a, 2005), R
eblov
a & Seifert
(2004), R
eblov
a (2006, 2013), Arzanlou et al. (2007),
Spatafora et al. (2007), Damm et al. (2008), Schoch et al.
(2009), Shenoy et al. (2010), R
eblov
aet al. (2011, 2015b,
2016), Jaklitsch et al. (2013) Untereiner et al. (2013),
Hern
andez-Restrepo et al. (2014), Su et al. (2016).
Sequence alignment
ITS, nucLSU, nucSSU and rpb2 sequences were manually
aligned in BioEdit v. 7.1.8 (Hall 1999). Alignments of act1 and
tub2 sequences were generated in MAFFT v. 7 (Katoh &
Standley 2013) and manually corrected where necessary.
Consensus 2D structure models for the ITS1 and ITS2 were
obtained for all members of the Xenospadicoidales and used to
determine positions of homologous nucleotides in the ITS
alignment. Introns occurring in nucLSU and nucSSU were
delimited manually and excluded from the alignment; in addition,
438 nucleotides (nt) of nucLSU at the 3
0
-end and 127 nt of
nucSSU at the 5
0
-end were excluded from the alignment
because of the incompleteness in the majority of sequences.
The single-locus data sets were examined for topological
incongruence among loci for members of the Xenospadicoidales
(act1: 35 sequences/338 characters including gaps, ITS: 37/757,
nucLSU: 39/1 842, nucSSU: 34/1 668, rpb2: 29/1 127, tub2: 28/
966), and members of the Sordariomycetidae (nucLSU: 104/
1 973, nucSSU: 71/1 787, rpb2: 65/1 189). Congruence among
the loci was tested using the 70 % reciprocal bootstrap criterion
(Mason-Gamer & Kellogg 1996). For each individual partition,
1 000 bootstrap replicates were generated with RAxML-HPC v.
7.0.3 (Stamatakis 2006) and PAUP v. 4.0b10 (Swofford 2002)
and compared visually for topological conflict among supported
clades in phylogenetic trees. The conflict-free alignments were
concatenated into a multi-locus alignment that was subjected to
subsequent phylogenetic analyses. The multiple sequence
alignment is deposited in TreeBASE (S21034).
Phylogenetic analyses
In order to explore monophyly and infrageneric relationships
within Lentomitella,Spadicoides,Torrentispora and other
morphologically similar taxa, and to resolve their phylogenetic
relationships in a broader context we performed analyses of
combined act1, ITS, nucLSU, nucSSU, rpb2 and tub2 sequences
on two datasets: a reduced dataset consisting of members of
these genera and a full dataset consisting of these taxa along
SORDARIOMYCETES
www.studiesinmycology.org 3
Table 1. List of fungi, isolate information and new sequences determined for this study and those retrieved from GenBank. The asterisk (*) denotes ex-type strains of members of the
Xenospadicoidales. GenBank accession numbers in bold were generated for this study.
Classification Taxon Source GenBank accession numbers
nucLSU nucSSU rpb2 ITS act1 tub2 References
Annulatascales Annulatascus velatisporus A 70-18 AY316354 – ––––Raja et al. (2003)
Annulusmagnus triseptatus CBS 131483 GQ996540 JQ429242 JQ429258 –––R
eblov
aet al. (2010, 2012)
Ascitendus austriacus CBS 131685 GQ996539 GQ996542 JQ429257 –––R
eblov
aet al. (2010, 2012)
Atractosporales Atractospora decumbens CBS 139032 KT991658 KT991640 KT991647 –––R
eblov
aet al. (2016)
A. ellipsoidea A 411-3 AY316356 – ––––Raja et al. (2003)
A. reticulata CBS 127884 KT991660 –KT991649 –––R
eblov
aet al. (2016)
A. verruculosa CBS 132040 KT991659 KT991641 KT991648 –––R
eblov
aet al. (2016)
Rubellisphaeria abscondita CBS 132078 KT991666 KT991646 KT991657 –––R
eblov
aet al. (2016)
Barbatosphaeriaceae Barbatosphaeria barbirostris CBS 121149 EF577059 KM492851 KM492903 –––R
eblov
aet al. (2015b)
B. dryina CBS 127691 KM492864 KM492852 KM492904 –––R
eblov
aet al. (2015b)
Boliniales Camarops microspora CBS 649.92 AY083821 DQ471036 DQ470937 –––Smith et al. (2003), Spatafora et al. (2007)
Apiorhynchostoma curreyi UAMH 11088 JX460989 KY931894 KY931926 –––Untereiner et al. (2013), this study
Endoxyla operculata UAMH 11085 JX460992 KY931895 KY931927 –––Untereiner et al. (2013), this study
Calosphaeriales Calosphaeria pulchella CBS 115999 AY761075 AY761071 GU180661 –––R
eblov
aet al. (2004, 2011)
Jattaea algeriensis CBS 120871 EU367457 EU367462 HQ878603 –––Damm et al. (2008), R
eblov
a (2011)
Togniniella acerosa CBS 113648 AY761076 AY761073 GU180660 –––R
eblov
aet al. (2004), R
eblov
a (2011)
Chaetosphaeriales Chaetosphaeria ciliata ICMP 18253 GU180637 GU180614 GU180659 –––R
eblov
aet al. (2011)
C. curvispora ICMP 18255 GU180636 AY502933 GU180655 –––R
eblov
aet al. (2011)
Melanochaeta hemipsila S.M.H. 2125 AY346292 –AY780184 –––Huhndorf et al. (2004), Miller & Huhndorf (2005)
Coniochaetales Barrina polyspora AWR 9560A AY346261 – – –––Huhndorf et al. (2004)
Coniochaeta discoidea SANK 12878 AY346297 AJ875179 AY780191 –––Huhndorf et al. (2004), García et al. (2006)
C. ostrea CBS 507.70 DQ470959 DQ471007 DQ470909 –––Spatafora et al. (2007)
Cordanales Cordana terrestris ICMP 15117 EF063573 – ––––R
eblov
a & Seifert (2007)
C. pauciseptata M.R. 1150 AF178563 – – –––R
eblov
a & Winka (2000)
C. pauciseptata CBS 113708 EF204507 –EF204490 –––Shenoy et al. (2010)
C. pauciseptata IMI 102120 HE672158 – – –––Hern
andez-Restrepo et al. (2014)
C. ellipsoidea IMI 229746 HE672156 – ––––Hern
andez-Restrepo et al. (2014)
C. inaequalis CBS 508.83 HE672157 – – –––Hern
andez-Restrepo et al. (2014)
Diaporthales Diaporthe phaseolorum FAU 458, NRRL 13736 U47830 L36985 AY641036 –––Spatafora & Blackwell (1993), Reeb et al. (2004)
Gnomonia gnomon CBS 199.53 AF408361 DQ471019 DQ470922 –––Castlebury et al. (2002), Spatafora et al. (2007)
Valsa ambiens AR 3516 AF362564 DQ862056 DQ862025 –––Zhang et al. (2007)
Distoseptisporaceae Distoseptispora fluminicola MFLUCC 15-0417 KU376270 – – –––Su et al. (2016)
D. aquatica MFLUCC 15-0374 KU376268 – – –––Su et al. (2016)
D. adscendens HKUCC 10820 DQ408561 –DQ435092 –––Shenoy et al. (2006)
D. leonensis HKUCC 10822 DQ408566 –DQ435089 –––Shenoy et al. (2006)
Jobellisiales Jobellisia fraterna S.M.H. 2863 AY346285 – – –––Huhndorf et al. (2004)
J. luteola S.M.H. 2753 AY346286 – – –––Huhndorf et al. (2004)
Magnaporthales Gaeumannomyces graminis AR 3401, M 57 AF362557 JF414874 ––––Farr et al. (2001), Zhang et al. (2011)
Macgarvieomyces borealis CBS 461.65 DQ341511 DQ341489 KM485070 –––Thongkantha et al. (2009), Klabauf et al. (2014)
R
EBLOV
AET AL.
4
Table 1. (Continued).
Classification Taxon Source GenBank accession numbers
nucLSU nucSSU rpb2 ITS act1 tub2 References
Magnaporthe grisea Ina168, 70-15 AB026819 DQ493955 ––––Sone et al. (2000), Rehmeyer et al. (2006)
Myrmecridiales Myrmecridium flexuosum CBS 398.76 EU041825 ––EU041768 ––Arzanlou et al. (2007)
M. montsegurinum PRM 934684 KT991664 KT991645 KT991654 KT991674 ––R
eblov
aet al. (2016)
M. schulzeri CBS 100.54 EU041826 ––EU041769 ––Arzanlou et al. (2007)
Ophiostomatales Ceratocystiopsis minuta UM 1533, WIN(M)1537 EU913657 HQ634854 ––––Plattner et al. (2009), Hafez et al. (2012)
Fragosphaeria purpurea CBS 133.34 AF096191 AF096176 ––––Suh & Blackwell (1999)
Ophiostoma piliferum CBS 158.74 DQ470955 DQ471003 DQ470905 –––Spatafora et al. (2007)
Raffaelea ambrosiae CBS 185.64 EU984297 AY497518 – –––Gebhardt et al. (2004), Massoumi Alamouti et al. (2009)
Papulosaceae Brunneosporella aquatica HKUCC 3708 AF132326 – – –––Ranghoo et al. (1999)
Fluminicola coronata HKUCC 3717 AF132332 – ––––Ranghoo et al. (1999)
Papulosa amerospora J.K. 5547F DQ470950 DQ470998 DQ470901 –––Spatafora et al. (2007)
Phomatosporales Lanspora coronata J.K. 4839A U46889 DQ470996 DQ470899 –––Spatafora et al. (1998, 2007)
Phomatospora bellaminuta J.K. 5543N FJ176857 FJ176803 FJ238345 –––Schoch et al. (2009)
Sordariales Gelasinospora tetrasperma CBS 178.33 DQ470980 DQ471032 DQ470932 –––Spatafora et al. (2007)
Lasiosphaeria ovina S.M.H. 1538, CBS 958.72 AF064643 AY083799 AY600292 –––Fern
andez et al. (1999), Smith et al. (2003),
Miller & Huhndorf (2004a)
Sordaria fimicola S.M.H. 4106, MUCL 937, CBS 723.96 AY780079 X69851 DQ368647 –––Miller & Huhndorf (2005), Tang et al. (2007)
Sporidesmiaceae Sporidesmium parvum HKUCC 10836 DQ408558 – ––––Shenoy et al. (2006)
S. minigelatinosa NN 47497 DQ408567 –DQ435090 –––Shenoy et al. (2006)
S. bambusicola HKUCC 3578 DQ408562 – ––––Shenoy et al. (2006)
S. fluminicola MFLUCC 15-0346 KU376271 – – –––Su et al. (2016)
S. aquaticum MFLUCC 15-0420 KU376273 – – –––Su et al. (2016)
S. submersum MFLUCC 15-0421 KU376272 – – –––Su et al. (2016)
Togniniales Phaeoacremonium minimum CBS 213.31, CBS 111015 AY761082 AY761068 HQ878610 –––R
eblov
aet al. (2004), R
eblov
a (2011)
P. fraxinopennsylvanicum CBS 128920 HQ878595 HQ878600 HQ878609 –––R
eblov
a (2011)
Woswasiaceae Woswasia atropurpurea CBS 133167 JX233658 JX233658 JX233659 –––Jaklitsch et al. (2013)
Xylochrysis lucida CBS 135996 KF539911 KF539912 KF539913 –––R
eblov
aet al. (2014)
Xenospadicoidales Calyptosphaeria collapsa PRA-12743* KY931834 KY931892 KY931861 KY931808 KY931771 KY931924 This study
C. subdenudata S.M.H. 3877* EU527994 ––
KY931774 KY931739 –Huhndorf et al. (2008), this study
C. subdenudata S.M.H. 2534 EU527993 ––KY931775 KY931740 –Huhndorf et al. (2008), this study
C. tenebrosa PRA-12742 –––KY931776 KY931741 –This study
C. tenebrosa PRA-12741 KY931809 KY931864 KY931836 KY931777 KY931742 KY931898 This study
C. tenebrosa PRA-12740* KY931810 KY931865 KY931837 KY931778 KY931743 KY931899 This study
C. tropica S.M.H. 1797* EU527992 KY931866 –KY931779 KY931744 –Huhndorf et al. (2008), this study
C. tropica S.M.H. 3225 EU527999 – ––––Huhndorf et al. (2008), this study
Lentomitella cirrhosa ICMP 15131* AY761085 AY761089 KM492911 KY931780 KY931745 KY931900 R
eblov
a (2006), R
eblov
aet al. (2015b), this study
L. crinigera CBS 138678 KY931811 KY931867 –KY931781 KY931746 KY931901 This study
L. conoidea CBS 131481 KT991663 KT991644 KT991653 KY931782 KY931747 –R
eblov
aet al. (2016), this study
L. conoidea CBS 131660 KY931812 KY931868 KY931840 KY931783 KY931748 KY931902 This study
L. conoidea CBS 141370 KY931813 KY931869 KY931841 KY931784 KY931749 KY931903 This study
(continued on next page)
SORDARIOMYCETES
www.studiesinmycology.org 5
Table 1. (Continued).
Classification Taxon Source GenBank accession numbers
nucLSU nucSSU rpb2 ITS act1 tub2 References
L. conoidea M.R. 3135 KY931814 KY931870 KY931842 KY931785 KY931750 KY931904 This study
L. magna ICMP 18371* KY931815 KY931871 KY931843 KY931786 KY931751 –This study
L. obscura CBS 137799 KY931816 KY931872 KY931844 KY931787 KY931752 KY931905 This study
L. obscura CBS 138735 KY931817 KY931873 –KY931788 KY931753 KY931906 This study
L. obscura CBS 138736* KY931818 KY931874 KY931845 KY931789 KY931754 KY931907 This study
L. striatella ICMP 18369* KY931819 KY931875 KY931846 KY931790 KY931755 KY931908 This study
L. sulcata ICMP 15124* AY761086 KY931876 KY931847 KY931791 KY931756 KY931909 This study
L. tenuirostris CBS 138734* KY931821 KY931877 KY931849 KY931792 KY931758 KY931910 This study
L. tenuirostris CBS 141371 KY931822 KY931878 KY931850 KY931793 KY931759 KY931911 This study
L. vestita PRA-12739 KY931820 KY931879 KY931848 KY931794 KY931757 –This study
Lentomitella sp. M.R. 2953 KY931823 KY931880 –KY931795 –KY931912 This study
Spadicoides atra CBS 489.77 EF204506 EF204521 EF204489 –––Shenoy et al. (2010)
S. bina CBS 137794 KY931824 KY931881 KY931851 KY931796 KY931760 KY931913 This study
S. fuscolutea CBS 141262 KY931825 KY931882 KY931852 KY931797 KY931761 KY931914 This study
S. fuscolutea CBS 141263 KY931826 KY931883 KY931853 KY931798 KY931762 KY931915 This study
S. hyalostoma CBS 131268 KY931827 KY931884 KY931854 KY931799 KY931763 KY931916 This study
S. hyalostoma CBS 137793 KY931828 KY931885 KY931855 KY931800 KY931764 KY931917 This study
S. hyalostoma CBS 138688 KY931829 KY931886 KY931856 KY931801 KY931765 KY931918 This study
S. hyalostoma CBS 139771 KY931830 KY931887 KY931857 KY931802 KY931766 KY931919 This study
S. iberica CBS 127864* KY853527 – – –––Hern
andez-Restrepo et al. (2017)
Torrentispora aquatica HKU(M) 17484* AY780365 – ––––Vijaykrishna et al. (2005)
T. biatriispora A 464-3 AY316352 –KY931858 KY931803 KY931767 KY931920 Raja et al. (2003), this study
T. calembola PRA-12744* KY931831 KY931888 –KY931804 KY931768 KY931921 This study.
T. dubia PRA-12746 KY931832 KY931889 –KY931805 KY931769 KY931922 This study
T. fibrosa ICMP 15147 EF577060 KY931890 KY931859 KY931806 KY931770 KY931923 This study
T. novae-zelandiae ICMP 18368* KY931833 KY931891 KY931860 KY931807 ––This study
Genera incertae sedis Brachysporium nigrum M.R. 1346 KT991662 KT991643 KT991652 –––R
eblov
aet al. (2016)
B. polyseptatum DAOM 231136 AY281102 – – –––R
eblov
a & Seifert (2004)
Bullimyces communis AF 281-3 JF775585 JF758617 – –––Ferrer et al. (2012)
Ceratostomella cuspidata ICMP 17629 FJ617558 KT991642 KT991651 KT991671 KY931772 –R
eblov
aet al. (2016), this study
C. pyrenaica CBS 129343 KY931835 KY931893 KY931863 KT991672 KY931773 –R
eblov
aet al. (2016), this study
Ceratolenta caudata CBS 125234 JX066704 JX066708 JX066699 –––R
eblov
a (2013)
Natantiella ligneola CBS 123470 FJ617556 HQ878598 HQ878605 –––R
eblov
aet al. (2016)
Platytrachelon abietis CBS 125235 JX066703 JX066707 JX066698 –––R
eblov
a (2013)
Rhamphoria delicatula M.R. 1396 AF261068 AF242267 KT991655 –––Winka (2000), R
eblov
aet al. (2016)
Rhodoveronaea varioseptata CBS 123473 FJ617560 JX066710 JX066700 –––R
eblov
a (2009,2013)
Spadicoides verrucosa CBS 128.86 EF204508 EF204522 – –––Shenoy et al. (2010)
Sporidesmium tropicale HKUCC 10838 DQ408560 – ––––Shenoy et al. (2006)
Thyridium vestitum AR 3872 AY544671 AY544715 DQ470890 –––Schoch et al. (2009), Spatafora et al. (2007)
Xylomelasma sordida CBS 116000 AY761087 AY761090 KY931929 –––R
eblov
a (2006), this study
Helotiales (outgroup) Leotia lubrica AFTOL 1 AY544644 L37536 DQ470876 –––Gargas & Taylor (1995), Spatafora et al. (2007),
Schoch et al. (2009)
Microglossum rufum AFTOL 1292 DQ470981 DQ257358 DQ470933 –––Spatafora et al. (2007)
R
EBLOV
AET AL.
6
with homologous sequences of representatives of the subclass
Sordariomycetidae. In order to resolve relationships among
Pseudodiplococcium,Spadicoides and Xenospadicoides, a third
analysis was based on a reduced ITS-nucLSU dataset of their
representatives, mainly due to the availability of only a nucLSU
sequence for Pseudodiplococcium ibericum.TwoCeratostomella
species, C. cuspidata and C. pyrenaica (Sordariomycetidae,
incertae sedis), and Leotia lubrica and Microglossum rufum
(Helotiales,Leotiomycetes) were used to root the individual trees
in the reduced and full analyses.
The combined datasets were partitioned into several subsets
of nucleotide sites, i.e. ITS, nucLSU, nucSSU, rpb2 and coding
and non-coding regions of act1 and tub2. Bayesian Inference
(BI) and Maximum Likelihood (ML) analyses were used to esti-
mate phylogenetic relationships. BI analyses were performed in
a likelihood framework as implemented in MrBayes v. 3.2.6
(Huelsenbeck & Ronquist 2001) through the CIPRES Science
Gateway v. 3.3 (http://www.phylo.org). For the BI approach,
MrModeltest2 v. 2.3 (Nylander 2008) was used to infer the
appropriate substitution model that would best fit the model of
DNA evolution. The following models were selected according to
the Akaike information criterion for partitions for which we
assumed rate heterogeneity: GTR+I+G for ITS, nucLSU, rpb2
and coding region of act1, GTR+G for nucSSU and coding region
of tub2 and HKY+I+G for non-coding regions of act1 and tub2.
Two Bayesian searches were performed using default parame-
ters. The B-MCMCMC analyses lasted until the average stan-
dard deviation of split frequencies was below 0.01 with trees
saved every 1 000 generations. The first 25 % of saved trees,
representing the burn-in phase of the analysis, were discarded.
The remaining trees were used for calculating posterior proba-
bilities (PP) of recovered branches. ML analyses were performed
with RAxML-HPC v. 7.0.3 with a GTRCAT approximation. Nodal
support was determined by non-parametric bootstrapping (BS)
with 1 000 replicates. Maximum Parsimony (MP) analyses con-
ducted with PAUP v. 4.0b10 (Swofford 2002) were used as
supplementary to ML analyses to evaluate congruence among
loci and topological variation of single-gene phylogenetic trees. A
heuristic search was performed with the stepwise-addition option
with 1 000 random taxon addition replicates and TBR branch
swapping. All characters were unordered and given equal
weight. Gaps were treated as missing data. Branch support was
estimated on the recovered topologies by performing a heuristic
search of 1 000 bootstrap replicates consisting of ten random-
addition replicates for each bootstrap replicate.
Prediction of 2D structure models of ITS of
Lentomitella
Predicting the 2D structure of the variable and rapidly evolving
ITS region is essential for constructing a reliable multiple
sequence alignment to compare nucleotides at homologous
positions (in helices and loops) while searching for non-
conserved co-evolving nucleotides which maintain base pair-
ing. Consensus 2D structure models for the ITS1 and ITS2 were
built using the PPfold program v. 3.0 (Sukosd et al. 2012) which
uses an explicit evolutionary model and a probabilistic model of
structures, and relies on multiple sequence alignment of related
RNA sequences. The obtained 2D consensus models created for
all members of Xenospadicoidales were further improved using
the program Mfold (Zuker 2003) and then adjusted manually if
necessary, based on comparison of homologous positions in the
multiple sequence alignment. The predicted 2D RNA structures
were obtained in a dot bracket notation and were visualized and
drawn using the program VARNA: Visualization Applet for RNA
(Darty et al. 2009). The final 2D model of ITS2, which was further
utilized in formulating taxonomic hypotheses, was processed
with CorelDRAW Graphics Suite X4.
We performed in-depth comparative analyses of ITS2 2D
structures of Lentomitella spp. We identified three types of
substitutions in the aligned ITS sequences. The compensatory
base changes (CBCs) occur when both nucleotides of a paired
site mutate, i.e. G=C 4C=G, A-U or U-A, while maintaining a
canonical base pair. The hemi-compensatory base changes
(hCBCs) inflict the change of a canonical base pair to a near-
canonical so called “wobble”base pair, i.e. G=C /G/U. The
non-compensatory base changes (non-CBC) involve the
replacement of a canonical pair or a wobble pair with any non-
canonical pair. While the CBCs and hCBCs are responsible for
maintaining the RNA helix arrangement, non-CBCs lead to its
disruption (Leontis et al. 2002). All existing substitutions among
Lentomitella species identified in the ITS2 were mapped onto the
predicted 2D structure of ITS2 of L. vestita, the generic type (ITS
sequence: KY931794).
RESULTS
Topological variation in single-gene, five- and
six-gene phylogenetic trees
We studied molecular phylogenies of the Xenospadicoidales
based on six nuclear markers in order to compare their phylo-
genetic utility. Three loci were shown to provide the highest
number of distinct alignment patterns (RAxML) and parsimony
informative characters (PAUP): rpb2 (556/459), tub2 (671/414)
and ITS (496/347), which is one or more times as much as
provided by nucLSU (318/168), act1 (255/151) and nucSSU
(181/65) loci.
Although topologies of gene trees were generally concordant,
there was some incongruence regarding the position of spec-
imen PRA-12743. This specimen is morphologically highly
similar to species for which a new genus Calyptosphaeria is
introduced below. PRA-12743 was shown either nested in the
strongly supported Calyptosphaeria clade in ITS (100 % ML BS)
and nucSSU (99) trees or as sister to Calyptosphaeria in the tree
based on tub2 (82). In addition, it was resolved on a separate
branch; at the base (100) of the Xenospadicoidales clade in the
act1 tree or at the base of the Lentomitella/Calyptosphaeria clade
without support in the nucLSU tree or supported in the rpb2 tree
(not shown). This internode received 92 % and 69 % bootstrap
support in the ML and MP analyses, respectively, in the rpb2
tree. The absence of tub2 and rpb2 sequences of
C. subdenudata and C. tropica may affect tree topologies based
on these genes. In all single-gene phylogenies, Lentomitella,
Spadicoides and Torrentispora were always resolved as strongly
supported monophyletic clades except in the nucSSU where
Spadicoides is paraphyletic with low statistical support.
Two ML and BI phylogenetic analyses were performed for
comparison based on a five-gene data set (excluding rpb2, re-
sults not shown) and a six-gene data set (Fig. 1). In the five-gene
tree inferred from ML analysis, PRA-12743 is placed within the
SORDARIOMYCETES
www.studiesinmycology.org 7
0.07
Lentomitella tenuirostris CBS 141371
Lentomitella sulcata ICMP 15124T
Torrentispora aquatica HKU(M)17484T
Lentomitella conoidea CBS 141370
Torrentispora calembola PRA-12744T
Lentomitella conoidea M.R. 3135
Lentomitella tenuirostris CBS 138734T
Calyptosphaeria tenebrosa PRA-12742
Calyptosphaeria collapsa PRA-12743T
Spadicoides fuscolutea CBS 141262
Spadicoides fuscolutea CBS 141263
Spadicoides hyalostoma CBS 139771
Lentomitella crinigera CBS 138678
Torrentispora biatriispora A 464-3
Calyptosphaeria subdenudata S.M.H. 2534
Lentomitella conoidea CBS 131660
Lentomitella conoidea CBS 131481
Lentomitella obscura CBS 137799
Calyptosphaeria tropica S.M.H. 1797T
Ceratostomella pyrenaica CBS 129343
Lentomitella obscura CBS 138736T
Spadicoides bina CBS 137794
Torrentispora novae-zelandiae ICMP 18368T
Spadicoides hyalostoma CBS 137793
Torrentispora fibrosa ICMP 15147
Lentomitella vestita PRA-12739
Spadicoides hyalostoma CBS 131268
Ceratostomella cuspidata ICMP 17629T
Lentomitella sp. M.R. 2953
Lentomitella magna ICMP 18371T
Calyptosphaeria tenebrosa PRA-12741
Spadicoides hyalostoma CBS 138688
Calyptosphaeria subdenudata S.M.H. 3877T
Spadicoides atra CBS 489.77
Calyptosphaeria tropica S.M.H. 3225
Lentomitella striatella ICMP 18369T
Lentomitella obscura CBS 138735
Lentomitella cirrhosa ICMP 15131T
Calyptosphaeria tenebrosa PRA-12740T
Torrentispora dubia PRA-12746
86/0.96
98/1.0
93/1.0
94/1.0
87/0.99
87/0.8
93/0.9
80/0.8
70/1.0
81/0.96
97/1.0
96/1.0
82/0.9
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*Ceratostomella
(outgroup)
Xenospadicoidales,Xenospadicoidaceae
Lentomitella
Calyptosphaeria
Torrentispora
Spadicoides
Fig. 1. Phylogenetic analysis of members of the Xenospadicoidales. Phylogram inferred from the act1-ITS-nucLSU-nucSSU-tub2-rpb2 sequences with ML analysis using a
GTRCAT model of evolution. An asterisk (*) indicates branches with ML BS = 100 %, PP values = 1.0. Branch support of nodes 70 % ML BS and 0.80 PP is indicated
above or below branches. ‘T’after the name indicates type strain. Taxa given in bold are type species of Calyptosphaeria,Lentomitella,Spadicoides and Torrentispora.
R
EBLOV
AET AL.
8
Calyptosphaeria clade (100) on a branch next to C. tenebrosa
without support. In the five-gene tree inferred from BI analysis,
PRA-12743 is shown at the basal position within a strongly
supported clade (1.0 PP) as sister to Calyptosphaeria (0.81). In
the combined six-gene analysis, the position of PRA-12743 basal
in the Calyptosphaeria clade (97/1.0) is strongly supported (100/
1.0), and thus this analysis supports its inclusion in
Calyptosphaeria.
Because the only incongruence among data sets based on
individual genes was the placement of PRA-12743 in the rpb2
tree inferred from ML analysis, the data sets were concatenated.
Combination of these six nuclear loci provided robust phyloge-
netic support for all genera, based on a mixture of the faster
evolving ITS region and non-coding regions of act1 and tub2 and
generally more preserved and slower evolving regions like
nucSSU.
The comparison of ITS sequences among the three species
of Calyptosphaeria and PRA-12743 indicated that the latter taxon
has the highest divergence among these species. While the
length of the ITS sequences of Calyptosphaeria spp. varies
between 553–569 nt, the ITS sequence of PRA-12743 (655 nt)
is longer by ca. 100 nt with the longest insertion in the helix III of
the ITS2.
Phylogeny
In the reduced analysis, 38 combined act1, ITS, nucLSU,
nucSSU, rpb2 and tub2 sequences were assessed for 24 spe-
cies in four genera of the Xenospadicoidales. The alignment
consisted of 6 698 characters including gaps and 2 491 distinct
alignment patterns (ML analysis). No topological conflicts
occurred between trees generated from ML and BI analyses; the
ML tree is shown in Fig. 1. The Xenospadicoidales (100 % ML
BS/1.0 PP) are resolved with four strongly supported subclades
that represent Lentomitella (100/1.0), Spadicoides (100/1.0),
Torrentispora (96/1.0) and the newly introduced Calyptosphaeria
(100/1.0). Lentomitella comprises 16 strains belonging to nine
species, of which five are newly introduced to science. Cera-
tostomella fuscolutea and Ceratostomella hyalostoma grouped in
the Spadicoides clade and therefore these two species are
combined in Spadicoides.Xenospadicoides atra, the type spe-
cies, formerly described as Spadicoides atra, is nested in the
Spadicoides clade and therefore is accepted in the latter genus.
Ceratostomella dubia,Fusoidispora aquatica,Pseudoannula-
tascus biatriisporus and two unknown torrentispora-like fungi
grouped with T. fibrosa, the type species, in a monophyletic clade
resulting in two new species and three new combinations in
Torrentispora. The new genus Calyptosphaeria is introduced for
Ceratostomella subdenudata,Lentomitella tropica and two
morphologically similar hitherto unknown fungi. They are shown
on a separate strongly supported branch unrelated to the core of
Lentomitella.
The full data set consisted of combined nucLSU, nucSSU and
rpb2 sequences of 102 members of the Sordariomycetidae. This
alignment consisted of 4 949 characters and 2 280 distinct
alignment patterns (ML analysis). The ML tree is shown in Fig. 2.
The BI and ML tree topologies differed in the position of
C. collapsa. In the ML analysis, the Calyptosphaeria clade with
C. collapsa at the basal position is shown to be monophyletic but
statistically unsupported, while in the BI analysis C. collapsa is
shown on a separate branch. Another topological difference lies
in the position of Sporidesmiaceae and Atractosporales clades
as sisters to Xenospadicoidales in the BI analysis. The Sordar-
iomycetidae contain three major clades (A‒C). Clade A (99/1.0)
includes orders Calosphaeriales,Diaporthales,Jobellisiales and
Togniniales. Clade B (100/1.0) comprises five orders including
the Boliniales,Chaetosphaeriales,Coniochaetales,Cordanales
and Sordariales. Clade C (91/0.93) contains orders Annula-
tascales,Atractosporales,Magnaporthales,Myrmecridiales,
Ophiostomatales and Phomatosporales, and five families
including Barbatosphaeriaceae,Distoseptisporaceae,Pap-
ulosaceae,Sporidesmiaceae and Woswasiaceae. The Xen-
ospadicoidales are shown as a strongly supported monophyletic
group (98/1.0) embedded in clade C and distantly related to
Ceratostomella represented by C. cuspidata and C. pyrenaica in
our phylogeny. Its closest relatives are members of the Atrac-
tosporales,Ophiostomatales,Papulosaceae and Spor-
idesmiaceae and several genera of non-stromatic perithecial
ascomycetes and dematiaceous hyphomycetes of uncertain
positions. The strain CBS 113708 of Cordana pauciseptata,
formerly misidentified as Spadicoides bina (Shenoy et al. 2010,
Hern
andez-Restrepo et al. 2017), is grouped within the Corda-
nales; for details see Discussion.
The third analysis consisted of combined ITS and nucLSU
sequences of representatives of Pseudodiplococcium,Spadi-
coides and Xenospadicoides. The ITS sequence of ex-type
strain CBS 127864 of P. ibericum (KY853465, Hern
andez-
Restrepo et al. 2017) was excluded from the analysis due to
suspected contamination; in the Blast search it shows 91 %
similarity with Cordyceps emeiensis (AJ309347) and 90 %
similarity with Hirsutella jonesii (KJ524687) of the Hypocreales.
The alignment for ML analysis consisted of 2 601 characters and
232 distinct alignment patterns. No topological conflicts occurred
between trees generated from ML and BI analyses; the ML tree
is shown in the Supplementary Fig. 1.Spadicoides is shown as a
highly supported clade (100/1.0) including S. bina, the type
species, X. atra (as S. atra) and two other Spadicoides species.
Pseudodiplococcium ibericum, the type species, is nested in the
Spadicoides clade, and therefore is synonymised with the latter
genus.
Consensus 2D structure of ITS2 of Lentomitella
The predicted consensus 2D structure of ITS2, modelled for the
type species L. vestita (Figs 3, 4), is folded into the common core
structure typical for Eukaryota, i.e. a ring structure with four main
helices I–IV, of which helices II (35 nt) and III (71 nt) are highly
conserved. The folding pattern of the last region corresponding to
helix IV is highly variable among Lentomitella spp.; in the case of
L. vestita it adopts a short helix. Therefore, only helices I –III were
evaluated.
Three CBCs on 7, 9 and 10th base pairs were identified in
helix I but only the CBC on a 10th base pair (C=G /U-A) is
unique between L. crinigera and other Lentomitella species. The
other two CBCs show a certain degree of homoplasy and
characterise several clades or single branches corresponding to
individual species. At the same position, the CBC was accom-
panied by either hCBC in two cases and by a non-CBC in a
single case. In addition, four hCBCs and two non-CBCs were
identified in helix I (Fig. 3).
In helix II no CBC was identified, only seven hCBCs and three
non-CBCs occur here (Fig. 4). The length of helix II is longer by
SORDARIOMYCETES
www.studiesinmycology.org 9
0.08
Lentomitella cirrhosa ICMP 15131
Spadicoides verrucosa CBS 128.86
Calyptosphaeria tropica S.M.H. 1797
Torrentispora calembola PRA-12744
Ceratostomella cuspidata ICMP 17629
Cordana ellipsoidea IMI 229746
Calyptosphaeria tenebrosa PRA-12740
Cordana inaequalis CBS 508.83
Lentomitella striatella ICMP 18369
Thyridium vestitum AR 3872
Xylomelasma sordida CBS 116000
Lentomitella sulcata ICMP 15124
Torrentispora novae-zelandiae ICMP 18368
Platytrachelon abietis CBS 125235
Torrentispora fibrosa ICMP 15147
Brachysporium nigrum M.R. 1346
Spadicoides fuscolutea CBS 141263
Lentomitella magna ICMP 18371
Lentomitella tenuirostris CBS 138734
Microglossum rufum AFTOL ID-1292
Cordana pauciseptata CBS 113708
Cordana pauciseptata IMI 102120
Natantiella ligneola CBS 123470
Lentomitella vestita PRA-12739
Spadicoides bina CBS 137794
Rhodoveronaea varioseptata CBS 123473
Sporidesmium tropicale HKUCC 10838
Spadicoides atra CBS 489.77
Torrentispora aquatica HKU(M) 17484
Ceratostomella pyrenaica CBS 129343
Cordana terrestris ICMP 15117
Rhamphoria delicatula M.R. 1396
Lentomitella crinigera CBS 138678
Spadicoides hyalostoma CBS 137793
Brachysporium polyseptatum DAOM 231138
Bullimyces communis AF 281-3
Cordana pauciseptata M.R. 1150
Calyptosphaeria collapsa PRA-12743
Calyptosphaeria subdenudata S.M.H. 3877
Torrentispora dubia PRA-12746
Lentomitella conoidea M.R. 3135
Ceratolenta caudata CBS 125234
Lentomitella obscura M.R. 3801
Torrentispora biatriispora A 464-3
96/1.0
99/1.0
79/0.84
99/1.0
97/1.0
77/1.0
79/0.94
99/1.0
98/1.0
88/1.0
98/1.0
90/0.98
93/0.99
84/1.0
83/1.0
90/1.0
99/1.0
79/1.0
98/1.0
91/0.93
84/0.84
84/-
88/0.99
79/1.0
99/1.0
93/1.0
97/1.0
94/1.0
88/1.0
77/-
83/1.0
*
*
-/1.0
*
*
*
*
Sporidesmiaceae
Papulosaceae
Atractosporales
*Ophiostomatales
*
*
*Barbatosphaeriaceae
Distoseptisporaceae
Magnaporthales
Phomatosporales
*
*
*Woswasiaceae
Myrmecridiales
Annulatascales
*
Coniochaetales
Boliniales
Chaetosphaeriales
Cordanales
Sordariales
Calosphaeriales
Jobellisiales
Togniniales
Diaporthales
*
*
**
*
*
*
*
**
*Leotia lubrica AFTOL ID-1
Lentomitella
Calyptosphaeria
Torrentispora
Spadicoides
Xenospadicoidales, Xenospadicoidaceae
-/1.0
-/1.0
-/1.0
70/0.99
Sordariomycetidae
as ‘Spadicoides bina’
Shenoy et al. (2010)
Leotiomycetes (outgroup)
Clade A
Clade B
Clade C
Fig. 2. Phylogenetic analysis of selected members of the Sordariomycetidae. Phylogram inferred from the nucLSU-nucSSU-rpb2 sequences with ML analysis using a GTRCAT
model of evolution. Details as in Fig. 1.
R
EBLOV
AET AL.
10
Fig. 3. ITS2 secondary structure of Lentomitella vestita (GenBank accession no. KY931794) and 5.8S-28S rRNA gene hybridization (proximal stem region) (above); detail of
helix I (below). ITS2 helices are numbered I–IV. All substitutions recorded among members of Lentomitella are mapped on the 2D model. Identified substitutions are colour-
coded: CBC (blue), hCBC (green) and non-CBC (red); position with all types of substitution (black). Parts of the text highlighted with grey colour refer to CBCs. Parts of hairpin
loops and a helix highlighted with grey colour represent regions with a variable number of nucleotides or sequence variation.
SORDARIOMYCETES
www.studiesinmycology.org 11
two base pairs in all Lentomitella species than in L. vestita.In
L. vestita the folding of the first two nucleotides at the 5
0
-end and
the two last nucleotides at the 3
0
-end has not been predicted
(leading to U-A, U/U base pairs), while in all other species of the
genus the folding pattern of the first two pairs is preserved and
nucleotides are conserved (base pairs U-A, U/G). The
pyrimidine-pyrimidine mismatch in helix II (Mai & Coleman 1997,
Schultz et al. 2005) was observed only in L. cirrhosa and
L. striatella (base pairs U/C, C/C).
Helix III is the longest with two asymmetrical loops, bulges
and a hairpin loop. It contains four CBCs on 8, 14, 16, and 24th
base pairs, but also an additional 11 hCBCs and two non-CBCs.
At the same position, CBC was accompanied by additional
substitutions: by hCBC in four cases and in two events by non-
CBC. Only the 14th base pair contained all types of substitutions
involving CBC, hCBC, and non-CBC and is valuable for studying
the evolution of CBC. The CBCs that characterise CBC clades
and correspond to biological species were identified for
L. magna,L. sulcata,L. vestita and one CBC delimits a clade
containing L. obscura,L. sulcata, and Lentomitella sp. Two pairs
of species that do not have CBCs between them but are sepa-
rated by a hCBC in L. cirrhosa (G/U) and L. striatella (G=C), and
by a non-CBC in L. conoidea (G/A) and L. tenuirostris (G=C) on
the 14th base pair, suggesting an unfinished segregation of these
species, are discussed below.
Taxonomy
Xenospadicoidales Hern.-Restr. et al., Stud. Mycol. 86: 91.
2017; emend. R
eblov
a & A.N. Mill.
Emended description: Lignicolous. Ascomata perithecial, non-
stromatic. Ostiole periphysate. Hamathecium of paraphyses.
Asci unitunicate, persistent, 8-spored, with a non-amyloid apical
annulus. Ascospores hyaline or pale brown prior to discharge,
aseptate or septate, variable in shape. Asexual morphs dema-
tiaceous hyphomycetes. Conidiophores macronematous, mon-
onematous. Conidiogenous cells tretic or holoblastic-denticulate,
sympodially proliferating. Conidia hyaline or brown, aseptate or
septate, variable in shape.
Type family:Xenospadicoidaceae Hern.-Restr. et al.
Xenospadicoidaceae Hern.-Restr. et al., Stud. Mycol. 86: 91.
2017; emend. R
eblov
a & A.N. Mill.
Synonym:Lentomitellaceae H. Zhang et al., Fungal Div. 85: 95.
2017.
Emended description: Lignicolous. Ascomata perithecial, non-
stromatic, with venter immersed, partially erumpent becoming
superficial. Neck cylindrical or rostrate with or without sulcations.
Ostiole periphysate. Hamathecium consisting of septate,
tapering paraphyses. Asci unitunicate, persistent, cylindrical or
cylindrical-clavate, 8-spored, with a non-amyloid apical annulus.
Ascospores hyaline or pale brown prior to discharge, aseptate or
septate, variable in shape, smooth-walled or ornamented.
Asexual morphs dematiaceous hyphomycetes producing effuse
colonies. Conidiophores macronematous, mononematous,
branched or unbranched. Conidiogenous cells tretic or
holoblastic-denticulate, sympodially proliferating. Conidia hyaline
or brown, aseptate or septate, variable in shape.
Type genus:Spadicoides S. Hughes (as Xenospadicoides Hern.-
Restr. et al., Stud. Mycol. 86: 92. 2017)
Key to genera accepted in the Xenospadicoidales
Calyptosphaeria R
eblov
a & A.N. Mill., gen. nov. MycoBank
MB821760
Etymology:Kalypt
o(Gr.) meaning hide, conceal or envelop,
referring to this taxon, which remained hidden within Lentomitella
until its position could be resolved with DNA sequence data.
Sexual morph:Ascomata perithecial, non-stromatic, immersed,
partially erumpent becoming superficial with only bases
immersed, scattered or grouped, varying in position from upright
to nearly horizontal; venter globose, subglobose to conical,
clothed with hairs. Neck conical, cylindrical or rostrate with 3–4
deep sulcations or roughened lacking sulcations, dark brown,
upright or slightly decumbent. Ostiole periphysate. Ascomatal
wall fragile, two-layered. Paraphyses becoming partially dis-
integrated, septate. Asci unitunicate, cylindrical, short-stipitate, 8-
spored; apex with a distinct, non-amyloid apical annulus. Asco-
spores ellipsoidal or ellipsoidal-fusiform, sometimes flattened on
one side, hyaline becoming pale brown or dull brown prior to
discharge, aseptate with a delayed formation of three transverse
septa when still within the ascus, smooth-walled, without sheath
or appendages. Asexual morph: unknown.
Type species:Calyptosphaeria tenebrosa R
eblov
a & A.N. Mill.
Notes:Huhndorf et al. (2008) expanded the generic concept of
Lentomitella by including two species with ellipsoidal or ellipsoidal-
fusiform, smooth-walled ascospores that turn brown prior to
discharge, i.e. L. pallibrunnea and L. tropica, and showed their
sister relationship to L. cirrhosa and L. crinigera using partial
nucLSU sequence data. Based on the combined analysis of six
nuclear markers and morphology of ascospores, the new genus
Calyptosphaeria is segregated from Lentomitella to accommodate
L. pallibrunnea and L. tropica and two other morphologically similar
taxa. The formation of septa is delayed and mature ascospores
remain mostly aseptate and, in some cases, visible but indistinct
cytoplasmic bands appear in areas where septa would be ex-
pected to form (Barr 1986 and this study). The ascospores are
uniseriate or overlapping uniseriate within the asci, although
sometimes they can be partially biseriate in the middle of the
sporiferous part and asci appear slightly clavate.
Key to the species accepted in Calyptosphaeria
1. Ascospores brown prior to discharge ……………………….Calyptosphaeria
1. Ascospores hyaline prior to discharge ……………………………………….. 2
2. Ascospores thick-walled, smooth-walled, ascomatal neck without sulcations
…………………………………………………………………….…Torrentispora
2. Ascospores thin-walled, smooth-walled or ornamented, ascomatal neck
with or without sulcations …………………………………………………….... 3
3. Ascospores longitudinally striate, conidiogenesis holoblastic-denticulate
………………………………………………………………………… Lentomitella
3. Ascospores smooth-walled or delicately verrucose, conidiogenesis tretic
and holoblastic-denticulate ……………………………………….. Spadicoides
1. Ascomatal neck roughened lacking sulcations, ascospores (10 –)
10.5–12( –12.5) × 4.5 –5μm……………………………………… C. collapsa
1. Ascomatal neck sulcate ………………………………………………………... 2
2. Ascospore longer than 16 μm, (16 –)17–20(–21) × 6–7μm……………….
……………………………………………………………..………… C. tenebrosa
SORDARIOMYCETES
www.studiesinmycology.org 13
Calyptosphaeria collapsa R
eblov
a & A.N. Mill., sp. nov.
MycoBank MB821761. Fig. 5.
Etymology:Collapsus (L.) meaning collapsed, referring to as-
cospores which collapse laterally upon aging.
Sexual morph:Ascomata immersed, partially erumpent
becoming superficial with only bases immersed, closely
grouped. Venter 350–490 μm diam, 400–500 μm high, sub-
globose, upright or sometimes lying horizontally on the host,
dark brown to black, with brown, septate hairs 2–3μmwide
sparsely covering the sides and bottom. Neck central,
120–140 μm wide, up to 700 μm long, cylindrical, upright, apex
roughened, without sulcations. Ostiole periphysate. Ascomatal
wall fragile to leathery, 44–62(–75) μm thick, two-layered; outer
layer consisting of thick-walled, brown, polyhedral cells of tex-
tura prismatica to textura epidermoidea with opaque walls; cells
tending to be darker towards the outside, more flattened and
paler towards the interior. Inner layer consisting of several rows
of thin-walled, hyaline, flattened cells. Paraphyses abundant,
longer than the asci, becoming disintegrated with age, septate,
slightly constricted at the septa, hyaline, 6.5–9.5 μm wide,
tapering to ca. 3 μm apically. Asci 82–96(–100) × (7–)
7.5–8.5(–9) μm (mean ± SD = 89.4 ± 5.3 × 8 ± 0.4 μm),
67–72(–86) μm (mean ± SD = 77.2 ± 5.1 μm) long in the
sporiferous part, truncate at the apex, cylindrical, with a short
stipe; with 8 overlapping uniseriate or partly biseriate asco-
spores; apical annulus 3.5–4μm wide, ca. 2.5 μm high. As-
cospores (10–)10.5–12(–12.5) × 4.5–5μm (mean ± SD = 11.3
± 0.6 × 4.8 ± 0.3 μm), ellipsoidal to fusiform, sometimes slightly
flattened on one side, often collapsing laterally upon aging,
aseptate, later with up to three indistinct cytoplasmatic bands,
pale brown prior to discharge, smooth-walled. Asexual morph:
unknown.
Fig. 5. Calyptosphaeria collapsa.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D. Paraphyses. E. Ascal apex with apical annulus. F–I. Asci. J,K. Ascospores.
A–K from PRA-12743. Scale bars: A, B = 500 μm, C = 20 μm, E, J, K = 5 μm, D, F–I=10μm.
2. Ascospores up to 16 μm long …………………………………………..…….. 3
3. Ascospores 11–14.5(–16) × 4–5μm…………………..….. C. subdenudata
3. Ascospores 14.5–16 × 5–6μm………………………………..…... C. tropica
R
EBLOV
AET AL.
14
Specimens examined:Czech Republic, Southern Moravia, Lednice, Nejdek old
Slavic settlement, area close to the pagan burial grounds, on decaying wood of
Carpinus betulus, 16 Nov. 2014, M. R
eblov
a M.R. 3881 (holotype, PRA-12743);
ibid., M.R. 3882, M.R. 3884.
Notes:Calyptosphaeria collapsa differs from other species of the
genus by a roughened ascomatal neck without sulcations. In the
absence of an asexual morph (Calyptosphaeria spp. did not
germinate in vitro) it is difficult to find any other morphological
differences between them. It resembles C. subdenudata and
C. tropica in the morphology of ascospores, but the ascospores
are shorter in C.collapsa. It further differs from C. subdenudata
in having longer asci and from C.tropica in having a cylindrical
neck vs. conical rostrate ascomatal apex in the latter species.
The collapsing ascospores were observed in water, lactic acid
and Melzer's reagent.
Calyptosphaeria collapsa was found on strongly decaying
wood of several fallen trunks of Carpinus betulus in the Czech
Republic, the remains of old growth trees that were more than
one hundred years old.
Calyptosphaeria subdenudata (Peck) R
eblov
a & A.N. Mill.,
comb. nov. MycoBank MB821762. Fig. 6.
Basionym:Sphaeria subdenudata Peck, Ann. Rep. N.Y. St. Mus.
nat. Hist. 32: 52. 1880 (1879).
Synonyms:Ceratostoma subdenudatum (Peck) Sacc., Syll. fung.
9: 481. 1891.
Ceratostomella subdenudata (Peck) M.E. Barr, Bull. N.Y. St.
Mus. 459: 44. 1986.
Wegelina subdenudata (Peck) M.E. Barr, Cryptog. Bryol.-
Lich
enol. 19: 172. 1998.
Lentomitella pallibrunnea Huhndorf et al., Mycologia 100: 948.
2008.
Xylomelasma moderata Lar.N. Vassiljeva & S.L. Stephenson,
Mycosphere 5: 223. 2014.
Sexual morph:Ascomata immersed, partially erumpent with
protruding necks becoming superficial with only bases
immersed, scattered or in groups. Venter 390–500 μm diam,
400–520 μm high, subglobose, upright, more often lying hori-
zontally on the host, dark brown to black, with reddish brown,
septate hairs ca. 3.5 μm diam sparsely covering the sides. Neck
central, 100–130 μm wide, up to 500 μm long, cylindrical, up-
right, slightly roughened, apex with several deep sulcations.
Ostiole periphysate. Ascomatal wall fragile, 37–40 μm thick, two-
layered; outer layer consisting of thick-walled, brown, polyhedral
cells of textura prismatica with opaque walls; cells tending to be
darker towards the outside, becoming flattened and paler to-
wards the interior. Inner layer consisting of several rows of thin-
walled, hyaline, flattened cells. Paraphyses sparse, becoming
disintegrated with age, septate, slightly constricted at the septa,
hyaline, 5–6μm wide. Asci (50–)60–85(–90) × (7.5–)8–10 μm
(mean ± SD = 72.5 ± 3.2 × 9 ± 0.4 μm), truncate to broadly
rounded at the apex, cylindrical, with a short stipe; with 8
overlapping uniseriate or biseriate ascospores; apical annulus
ca. 3 μm wide, 2.5–3μm high. Ascospores 11 –14.5(–16) × 4–5
μm (mean ± SD = 13.3 ± 1.4 × 4.7 ± 0.4 μm), ellipsoidal, often
slightly flattened on one side and slightly curved, aseptate or with
several indistinct cytoplasmatic bands, hyaline becoming light
dull brown, smooth-walled. Asexual morph: unknown.
Fig. 6. Calyptosphaeria subdenudata.A–C. Ascomata. D, E. Longitudinal section of the ascomal wall. F. Ascospores. G, H. Ascal apex with apical annulus. I. Paraphyses.
J–M. Asci. A–M from JF 16082. Scale bars: A–C = 500 μm, D = 250 μm, E = 20 μm, F–H=5μm, I–M=10μm.
SORDARIOMYCETES
www.studiesinmycology.org 15
Specimens examined:French West Indies, Martinique, Sainte-Marie, La Phil-
ippe, coastal mesophilic rainforest, on decaying wood, 3 Aug. 2016, J. Fournier
J.F. 16082. USA, New York, Delaware County, Griffins', Catskill Mts., on decaying
wood, Sep. 1877, C.H. Peck (holotype of Sphaeria subdenudata, NYS); ibid.,
Indian Lake, on decaying wood, Oct. 1878, C.H. Peck (NYS). Michigan. Berrien
County, Warren Woods, south end of trail, through picnic area, up to creek, on
12 cm thick branch on the ground, 8 Nov. 1998, F.A. Fern
andez & A.N. Miller,
S.M.H. 3877 (holotype of Lentomitella pallibrunnea, F).
Notes:Calyptosphaeria subdenudata is known from French
West Indies (Martinique) and North America (Illinois, Massa-
chusetts, Michigan and New York), based on recently collected
material and the revision of the holotype and other herbarium
material of Lentomitella pallibrunnea (Huhndorf et al. 2008) and
Sphaeria subdenudata (Peck 1879, Barr 1986). Xylomelasma
moderata (Vassiljeva & Stephenson 2014)fits well within the
species concept based on the ascoma, ascus and ascospore
morphology. Both latter species are placed in synonymy under
C. subdenudata.Barr (1986) revised Peck's material of
S. subdenudata and noted that ascospores become 1–5-
pseudoseptate at maturity. Up to three cytoplasmic bands
were observed in areas where septa would be expected to form
during our revision of the type material of S. subdenudata; the
paraphyses were already disintegrated. The ascospores of
L. pallibrunnea were described as aseptate (Huhndorf et al.
2008).
Calyptosphaeria subdenudata is similar to C. collapsa in the
ellipsoidal, brown ascospores, but it differs in having shorter asci,
longer ascospores and a neck with deep sulcations.
Calyptosphaeria tenebrosa R
eblov
a & A.N. Mill., sp. nov.
MycoBank MB821763. Fig. 7.
Etymology:Tenebrosus (L.) meaning dark, referring to the colour
of the ascospores.
Sexual morph:Ascomata immersed, partially erumpent with
protruding necks or becoming superficial with only bases
immersed, scattered or grouped sometimes in rows. Venter
310–550 μm diam, 390–650 μm high, subglobose to conical,
straight or more often lying horizontally on the host, dark brown
to black, with brown, septate hairs 4.5–5μm diam sparsely
covering the lower part. Neck central, 120–140 μm wide, up to
600 μm long, cylindrical, upright, straight, often roughened, apex
slightly widened with 3–4 deep sulcations. Ostiole periphysate.
Ascomatal wall fragile to leathery, 55–65 μm thick, two-layered;
outer layer consisting of thick-walled, brown, polyhedral cells of
textura prismatica to textura angularis with opaque walls; cells
tending to be darker towards the outside, becoming flattened and
paler towards the interior. Inner layer consisting of several rows
of thin-walled, hyaline, flattened cells. Paraphyses abundant,
longer than the asci, becoming disintegrated with age, septate,
slightly constricted at the septa, hyaline, 5.5–9.5 μm wide,
tapering to 2.5–3.5 μm. Asci (119–)122–150 × 9–10(–11) μm
(mean ± SD = 134.7 ± 8.5 × 9.5 ± 0.3 μm), (105–)110–130 μm
(mean ± SD = 116.7 ± 6.4 μm) long in the sporiferous part,
truncate at the apex, cylindrical, with a short stipe; with 8 uni-
seriate or partly biseriate ascospores; apical annulus 4.5 –5μm
Fig. 7. Calyptosphaeria tenebrosa.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D. Paraphyses. E. Ascal apex with apical annulus. F–H. Ascospores. I–K.
Asci. A, D, J, K from PRA-12742, C, G, F, I from PRA-12741, B, E, H from PRA-12740. Scale bars: A, B = 500 μm, C = 20 μm, D, I–K=10μm, E–H=5μm.
R
EBLOV
AET AL.
16
wide, 4–4.5 μm high. Ascospores (16–)17–20(–21) × 6–7μm
(mean ± SD = 18.6 ± 1.1 × 6.4 ± 0.3 μm), ellipsoidal-fusiform,
tapering towards the ends, rarely inequilateral, sometimes
collapsing laterally upon aging, aseptate, three transverse septa
sometimes present in old ascospores, hyaline to yellowish
becoming pale brown prior to discharge, smooth-walled. Asexual
morph: unknown.
Specimens examined:Czech Republic, Central Bohemia, K
rivokl
atsko Pro-
tected landscape area, Ni
zbor, Vůznice Nature reserve, on decaying wood of
C. betulus, 20 Oct. 2004, M. R
eblov
a M.R. 2923. Northern Moravia, Podho
rí
‘Podhorn’near Hranice, on decaying wood of Fagus sylvatica, Mar. 1914, F.
Petrak (as Ceratostoma operculatum, Fl. Bohem. Morav. Exs. No. 968, PRM
655798). Southern Moravia, Lednice, Nejdek, forested area on the right side of
the Nejdek crossroads, on decaying wood of Acer campestre, 15 Nov. 2014, M.
R
eblov
a M.R. 3871 (holotype, PRA-12740); ibid., M.R. 3869; ibid., Nejdek, old
Slavic settlement near river Z
ameck
a Dyje, on decaying wood of Carpinus
betulus, 27 Oct. 2014, M. R
eblov
a M.R. 3867 (PRA-12741); ibid., Valtice,
Rendez-vous National nature monument, on decaying wood of Quercus cerris,
18 Nov. 2012, M. R
eblov
a M.R. 3704. France, Ari
ege, Lescure, Bois du Pas du
Baup, 500 m. a.s.l., on decaying wood of Alnus glutinosa, 24 Feb. 2004, J.
Fournier J.F. 04021 (PRA-12742). New Zealand, Westlands, Victoria Forest
Park, Rough Creek Road, 4 km S of Inangahua, on decaying wood, 22 Apr. 2005,
M. R
eblov
a M.R. 2967/NZ 810.
Notes:Calyptosphaeria tenebrosa can be distinguished from
other species of the genus by longer asci with the largest apical
annulus known in the genus and longer ellipsoidal-fusiform as-
cospores. It occurs on decaying wood of various hardwood trees
and it is known from several localities in the Czech Republic,
France and New Zealand.
Calyptosphaeria tropica (Huhndorf et al.)R
eblov
a & A.N. Mill.,
comb. nov. MycoBank MB821764.
Basionym:Lentomitella tropica Huhndorf et al., Mycologia 100:
948. 2008.
Notes: For description and illustration see Huhndorf et al. (2008).
Calyptosphaeria tropica can be distinguished from other species
of the genus by size of asci and ascospores, conical rostrate
ascomatal apex and tropical distribution (Costa Rica and Puerto
Rico).
Lentomitella Höhn., Annls mycol. 3: 552. 1906.
Sexual morph:Ascomata perithecial, non-stromatic, immersed or
partially erumpent with protruding necks or becoming superficial,
solitary or in rows or groups. Venter globose to subglobose,
clothed by brown or reddish-brown hairs. Neck central, cylin-
drical, upright to slightly decumbent, glabrous, apex sulcate.
Ostiole periphysate. Ascomatal wall fragile to leathery, two-
layered. Paraphyses longer than the asci, becoming dis-
integrated with age, septate, constricted at the septa, hyaline.
Asci cylindrical to clavate, broadly rounded or truncate at the
apex, with a short stipe; with 8 uniseriate or obliquely uniseriate
or overlapping, sometimes partly biseriate ascospores; with a
distinct, non-amyloid apical annulus. Ascospores ellipsoidal,
suboblong or ellipsoidal-fusiform, sometimes inequilateral, hya-
line, longitudinally striate, 1–3-septate with 2–4 large drops, with
a delayed formation of second and third septum. Asexual morph:
A phaeoisaria-like asexual morph is sometimes formed in vitro.
Conidiophores macronematous, mononematous, unbranched or
branched apically, arising from aerial hyphae, brown near the
base, subhyaline to hyaline towards the tip. Conidiogenous cells
terminal or intercalary, hyaline, subcylindrical to slender flask-
shaped, with a rachis bearing minute denticles, conidiogenesis
holoblastic-denticulate, conidial heads slimy, inconspicuous,
transparent. Conidia globose, ellipsoidal, clavate to obovate,
slightly apiculate at the base, hyaline, aseptate, smooth-walled.
Type species:Lentomitella vestita Höhn.
Notes:Lentomitella forms a strongly supported monophyletic
clade in the phylogenetic analysis based on six nuclear markers.
It is well-distinguished from other genera of the Xen-
ospadicoidales by hyaline, septate, longitudinally striate asco-
spores, distinct but relatively small apical annulus (2.5–3μm
wide, 1.5–2μm high) and ascomata with a cylindrical neck with
usually 3–4 deep sulcations at the apex and venter clothed by
dark interwoven hairs that can disappear upon aging. Lentomi-
tella comprises 11 species; L. cirrhosa,L. crinigera,
L. unipretoriae and L. vestita are accepted in the genus, two new
combinations are proposed for L. conoidea and L. investita, and
five species are described as new to science, i.e. L. magna,
L. obscura,L. sulcata,L. striatella and L. tenuirostris. The
phaeoisaria-like asexual morph has been experimentally proven
only for L. investita,L. sulcata, and Lentomitella sp. The asco-
spores do not germinate easily. Germinating tubes appear in ca.
1–2 wk after isolation on water agar. Despite the lack of known
asexual morphs, we observed that the colonies of individual
species differ macroscopically at the margin. The margin con-
sists either of densely branched hyphae, i.e. in L. conoidea,
L. crinigera and L. magna, or unbranched or sparsely branched
hyphae of the substrate mycelium in the other species.
Key to the species accepted in Lentomitella
1. Ascospores shorter than 11 μm…..…………………………………….….2
1. Ascospore longer than 11 μm…………………………………………….... 8
2. Ascospores 1-septate; (5.5–)6–7×3–3.5 μm………………….L. vestita
2. Ascospores 1–3-septate, longer than 7 μm………………………...…….3
3. Asci (46–)50–56(–65) × 6–7μm, ascospores (7 –)7.5–9 × 3.5–4(–4.5)
μm………………..………………………………………………... L. investita
3. Asci longer than 56 μm………………..…………………………………….4
4. Ascospores usually 1–2-septate, old ascospores 3-septate …………… 5
4. Ascospores 3-septate early in ontogeny ………………………………… 7
5. Ascospores 1(–3)-septate, first-formed septum in the middle,
8–10 × 4.5–5μm…………………………………………..…… L. cirrhosa
5. Ascospores usually 1–2-septate, first-formed septum in the middle or
slightly above or below the middle ………………………………………… 6
6. Ascospores up to 9.5 μm long; aerial mycelium on MLA woolly, woolly-
floccose or almost cobwebby at the margin; margin of the colony
consisting of densely branched hyphae ……………………….L. conoidea
6. Ascospores usually slightly longer, 9–10.5 μm; aerial mycelium on MLA
cottony to felty having a more compact appearance; margin of the colony
consisting of unbranched or sparsely branched hyphae ….. L. tenuirostris
7. Ascospores (8–)9–10.5(–11) × 4–5μm, asci 72 –81 × 6.5–7.5 μm……
……….………………………………………………………… L. unipretoriae
7. Ascospores longer than 10.5 μm………………………………………… 8
8. Ascospores (12.5–)13–15 × 5.5–6.5 μm………….…………… L. magna
8. Ascospores shorter than 13 μm…………………………………………… 9
SORDARIOMYCETES
www.studiesinmycology.org 17
Lentomitella cirrhosa (Pers.: Fr.) R
eblov
a, Mycologia 98: 82.
2006. Figs 8, 9.
Basionym:Sphaeria cirrhosa Pers., Syn. Meth. Fung. p. 59. 1801 :
Fries, Syst. Mycol. 2: 475. 1823.
Synonyms:Ceratostoma cirrhosum (Pers.: Fr.) Fuckel, Jahr.
nassau. Ver. Naturk. 23–24: 127. 1870.
Cerastoma cirrhosum (Pers.: Fr.) Qu
el., M
em. Soc.
Emul.
Montb
eliard, S
er. 2 5: 522. 1875.
Fig. 8. Lentomitella cirrhosa.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E, F. Asci. G. Paraphyses. H. Ascal apex with apical annulus. I–K. Ascospores. L,
M. Colonies on MLA and PCA after 28 d. A–M from ICMP 15131. Scale bars: A–C = 500 μm, D, G = 20 μm, E, F = 10 μm, H–K=5μm.
9. Preferring coniferous wood; ascomata up to 600 μm diam with a neck up
to 170 μm wide; ascospores (10–)10.5–13 × 4 –5.5 μm, asci (66 –)
70–88( –90) × (7.5 –)8–9.5(–10) μm…………..……………... L. crinigera
9. Species without substrate preferences, ascomata and necks smaller ……
……………………………………………………………………………….. 10
10. Asci up to 8 μm wide, ascospores up to 5 μm wide; asci (70–)
72–80( –82) × 7.5 –8(–8.5) μm, ascospores (10–)11–12 × 4 –5( –5.5)
μm……………………………….………………………………… L. obscura
10. Asci wider than 8 μm, ascospores wider than 5 μm…….……………… 11
11. Asci length/width ratio 9.4, ascospores (10.5–)11 –12( –13) μm long
………………………………………………………..……………... L. sulcata
11. Asci length/width ratio 8.6, ascospores 11.5–13(–14) μm long
……………………………………………………………………... L. striatella
R
EBLOV
AET AL.
18
Ceratostomella cirrhosa (Pers.: Fr.) Sacc., Michelia 1: 370. 1878.
Amphitrichum cirrhosum (Pers.: Fr.) Kuntze, Revis. gen. pl. 3(2):
443. 1898.
Endoxyla cirrhosa (Pers.: Fr.) Arx & E. Müll., Beitr. Krypt-Fl.
Schweiz 11(1): 355. 1954.
Sexual morph:Ascomata immersed to partially erumpent
becoming superficial, solitary or in small groups. Venter
300–330 μm diam, 300–340 μm high, globose to subglobose,
dark brown to black, covered with sparse, dark brown, septate
hairs ca. 3.5 μm wide. Neck central, 90–100 μm wide, up to
500 μm long, cylindrical, upright, straight or slightly flexuous,
glabrous, tapering, apex sulcate. Ostiole periphysate. Ascomatal
wall fragile to leathery, 25–40 μm thick, two-layered. Outer layer
consisting of thick-walled, brown, polyhedral cells of textura
prismatica with opaque walls; cells tending to be darker towards
the outside, becoming paler towards the interior. Inner layer
consisting of several rows of thin-walled, hyaline, flattened cells.
Paraphyses abundant, longer than the asci, becoming dis-
integrated with age, septate, constricted at the septa, hyaline,
7–10 μm wide, tapering to ca. 2.5–3μm. Asci (62–)
64–70(–73) × 6.5–7.5(–8.5) μm (mean ± SD = 66.7 ± 3.0 × 7.1
± 0.4 μm), (55–)58–62(–66) μm (mean ± SD = 60.2 ± 2.8 μm)
long in the sporiferous part, broadly rounded at the apex, cy-
lindrical to clavate with a short stipe; with 8 uniseriate or obliquely
uniseriate ascospores; apical annulus ca. 2.5 μm wide, 1.5 μm
high. Ascospores 8–10 × 4.5–5μm (mean ± SD =
9.4 ± 0.5 × 4.6 ± 0.3 μm), ellipsoidal, hyaline, longitudinally
striate, 1–3-septate, usually 1-septate with two large drops
becoming 3-septate upon aging. Asexual morph: unknown.
Culture characteristics: Colonies on MLA 16–19 mm diam after
14 d (19–23 mm after 21 d, 20–25 after 28 d) at 25 °C, circular,
convex. Aerial mycelium abundant, cottony, margins filamentous,
colony surface pale brown to cocoa brown; reverse black. Col-
onies on PCA 10–12 mm diam after 14 d (13–15 mm after 21 d,
14–16 mm after 28 d) at 25 °C, circular, convex. Aerial mycelium
abundant, woolly, loose to cobwebby towards the margin, colony
surface beige-grey with olive-brown inner ring and conspicuous
dark brown marginal ring of submerged hyphae; reverse black.
Vegetative hyphae branched, septate, medium to dark brown,
smooth-walled. Margin of the colony consisting of unbranched or
sparsely branched hyphae of substrate mycelium. Sporulation
not observed.
Lectotype; designated by R
eblov
a (2006): (Illustration) Persoon,
Icones pictae specierum rariorum Fungorum in Synopsi
Methodica Descriptarum. fasc. 4, tab. 24, fig. 3. 1808.
Specimens examined:New Zealand, South Island, Southland, Southland Distr.,
Fiordland National park, Kaherekoau Mts., Blue cliffs, Hump track 15 km W of
Papatotara, on decaying wood, 12 Mar. 2005, M. R
eblov
a M.R. 2952/NZ 481;
ibid., West Coast, Westland Distr., Hokitika, Mananui Point, Lake Mahinapua,
Swimmers Beach walk, on decaying wood of Podocarpus totara, 5 Mar. 2003, M.
R
eblov
a M.R. 2705/NZ 206 (epitype, PDD 81434, culture ex-epitype ICMP
15131).
Notes: It is challenging to interpret L. cirrhosa in the absence of a
type specimen in Persoon's herbarium (L). Persoon (1801)
described Sphaeria cirrhosa just with a few words. The illustra-
tion (Persoon 1808) showing black ascomata with immersed
venter clothed with sparse hairs and emerging glabrous neck
represents the only original element left and is reproduced here
as Fig. 9; the figure caption of tab. 24 in Persoon (1808)
incorrectly refers to pictures 1, 2 but S. cirrhosa is depicted in
picture 3. In the absence of a type specimen, this illustration
(Persoon 1808) was selected as lectotype [R
eblov
a 2006,
incorrectly cited as Persoon (1800), corrected here], and in order
to stabilise the species concept of L. cirrhosa, a collection from
New Zealand on decayed wood of Podocarpus totara (PDD
81434) was designated as epitype (R
eblov
a 2006).
Fuckel (1870) accepted S. cirrhosa in the genus Ceratostoma
as C. cirrhosum and provided a description based on his own
material published in Fungi Rhen. Exs. No. 1804 [= Lentomitella
vestita, this study]. Saccardo (1882) interpreted this species based
on various collections. He transferred S. cirrhosa to Ceratosto-
mella and described the ascomata as sparse, ca. 400 μm diam,
immersed becoming partially erumpent, subglobose, clothed with
hairs, with a long glabrous neck, asci cylindrical-clavate
65–75 × 7–9μm, containing eight ellipsoidal to oblong, hyaline
ascospores 9–12 × 3.5 μm, with 1–4, mostly two drops.
The species concept of Ceratostomella cirrhosa presented by
von Arx (1952) included 12 species synonyms and was based on
a revision of type and other herbarium material from Europe and
North America. The synonymy was partly adopted by R
eblov
a
(2006) and C. cirrhosa was transferred to the reinstated Lento-
mitella. The species description of L. cirrhosa was based on the
epitype, other herbarium material collected in Europe and on the
revision of holotypes of Sphaeria investita,Ceratostomella ves-
tita var. vestita and var. varvicensis and Eriosphaeria conoidea.
Fig. 9. Lentomitella cirrhosa.A. Description of Sphaeria cirrhosa (Persoon 1801). B. Illustration (lectotype) of S. cirrhosa (Persoon 1808).
SORDARIOMYCETES
www.studiesinmycology.org 19
In the light of phylogenetic analysis of six nuclear markers and
re-evaluation of morphological characters of ascospores and
asci, they are not conspecific with L. cirrhosa and belong to three
different species recognised in this study as L. conoidea,
L. investita (including C. vestita var. varvicensis as its synonym)
and L. vestita.
Although L. cirrhosa and L. vestita differ significantly in the
size of asci and ascospores, and the ascospores of the latter
species are regularly 1-septate vs. 1–3-septate in L. cirrhosa,
von Arx (1952) regarded them as conspecific. He considered the
holotype of L. vestita to be insufficiently developed and therefore
ignored the smaller size of ascospores (5.5–)6–7×3–3.5 μm
and asci 43–47(–55) × 5.5–6μm in his description of
C. cirrhosa [asci 50–80 μm long in the sporiferous part with stipe
20–40 μm long, ascospores 8–10 × 3.5 μmfide von Arx (1952)].
Another eight Ceratostomella species synonymised by von Arx
(1952) with L. cirrhosa, and previously transferred to Amphi-
trichum by Kuntze (1898), were revised; they belong to the
genera Ceratostomella (R
eblov
a 2006), Natantiella (R
eblov
a&
St
ep
anek 2009) and other fungi discussed below.
The ascospores of L. cirrhosa in the epitype (PDD 81434) are
mostly 1-septate with two large drops, but old ascospores
released from the asci possess three septa and four drops. The
closest relative to L. cirrhosa is L. striatella, which differs by
longer and wider asci and slightly longer and wider, regularly 3-
septate ascospores.
Lentomitella cirrhosa is also similar to L. investita and
L. conoidea in morphology of ascospores. It shares with
L. investita mostly 1-septate ascospores with the first-formed
septum positioned always in the middle and a delayed formation
of two additional septa. Lentomitella investita differs from
L. cirrhosa by shorter and narrower asci and slightly smaller as-
cospores. Lentomitella conoidea differs from L. cirrhosa by slightly
longer, mostly 1–2-septate ascospores with the first septum
formed in the middle or slightly above or below the middle.
Lentomitella conoidea (Feltg.) R
eblov
a, comb. nov. MycoBank
MB821765. Fig. 10.
Basionym:Eriosphaeria conoidea Feltg., Vorstud Pilzfl. Luxemb.,
Nachtr. 3: 282. 1903.
Sexual morph:Ascomata immersed to partially erumpent
becoming superficial, solitary or in groups. Venter 350–400 μm
diam, 370–450 μm high, globose to subglobose, dark brown to
black, covered by dark brown to reddish brown, septate hairs
3–4μm wide. Neck central, 100–110 μm wide, up to 800 μm
long, cylindrical, upright, straight or slightly flexuous, glabrous,
tapering, apex sulcate. Ostiole periphysate. Ascomatal wall
leathery, 45–57 μm thick, two-layered; outer layer consisting of
thick-walled, brown, polyhedral cells of textura angularis to tex-
tura prismatica with opaque walls; cells tending to be darker
towards the outside, becoming flattened and paler towards the
interior. Inner layer consisting of several rows of thin-walled,
hyaline, flattened cells. Paraphyses abundant, longer than the
asci, becoming disintegrated with age, septate, constricted at the
septa, hyaline, 5–9μm wide, tapering to ca. 3.5 μm. Asci
62–70(–75) × 6.5–7.5 μm (mean ± SD = 67.3 ± 2.9 ×
7.1 ± 0.2 μm), 58–63(–66) μm (mean ± SD = 61.3 ± 2.3 μm)
long in the sporiferous part, truncate at the apex, subcylindrical to
clavate, with a short stipe; with 8 partially overlapping, obliquely
uniseriate or partly biseriate ascospores; apical annulus ca.
2.5 μm wide, 1.5 μm high. Ascospores 8.5–9.5(–10.5) ×
4–4.5 μm (mean ± SD = 9.0 ± 0.5 × 4.5 ± 0.2 μm), ellipsoidal to
oblong, hyaline, longitudinally striate, 1–3-septate, commonly
only 1–2-septate, the first-formed septum in the middle or slightly
below or above the middle, third septum developing rarely.
Asexual morph: unknown.
Culture characteristics:ColoniesonMLA13–15 mm diam after
14 d (18–21 mm after 21 d, 24–25 mm after 28 d) at 25 °C, cir-
cular, convex. Aerial mycelium abundant, woolly to cottony
becoming loose to woolly-floccose at the margin, colony surface
brown-grey turning brown towards the margin; reverse black.
Colonies on PCA 8–9 mm after 14 d (10 –11 mm after 21 d, 10–12
after 28 d) at 25 °C, circular or slightly irregular, raised. Aerial
mycelium abundant, woolly to cottony, loose or almost cobwebby
towards the margin, colony surface brown-grey at the centre
turning dark olive-brown towards the margin; reverse black.
Vegetative hyphae branched, septate, medium to dark brown,
smooth-walled. Margin of the colony consisting of densely
branched hyphae of substrate mycelium. Sporulation not observed.
Specimens examined:Belgium, Moersdorf, on bark of Pyrus communis, 25 May
1902, J. Feltgen (holotype of Eriosphaeria conoidea, LUX 043455). Czech
Republic, Southern Bohemia,
Sumava Mts. National park, Sto
zec, Medv
edice
Mt., on decaying deciduous wood, 17 Sep. 2007, M. R
eblov
a M.R. 2998 (culture
CBS 141370). Denmark,Sjælland, Jægerspris Nordskov, distr. 44, on very rotten
wood of Quercus sp., 4 Nov. 1963, A. Munk (C); ibid., Ermelunded, on old bark of
Fraxinus excelsior, 7 Dec. 1964, A. Munk (C); ibid., Bernstorffsparken, on very
rotten wood, 24 Mar. 1965, A. Munk (C); ibid., Suserup Skov, on wood of Sorbus
sp., 15 Dec. 1995, T. Læssøe TL-4062 (C 34511). Italy, Lazio, province Viterbo,
Farnese, Selva di Lamone Nature reserve, on decaying wood of Quercus cerris,
19 Mar. 2010, M. R
eblov
a M.R. 3135; ibid., 3 Apr. 2011, M.R. 3607 (culture CBS
131660), M.R. 3611 (culture CBS 131481).
Notes: The examination of the holotype of Eriosphaeria conoidea
(Feltgen 1903) revealed a fungus that fits well the description of
Lentomitella and represents a distinct species. Therefore, it is
transferred to this genus and a new combination is proposed.
Höhnel (1906b) examined the holotype and concluded that
E. conoidea is conspecific with Ceratostomella debaryana
(Saccardo 1882) and that it shows remarkable similarity to
L. vestita and L. investita. Although C. debaryana shares similar
size of asci and ascospores with L. conoidea, in the protologue
(Auerswald 1869) the ascospores are described and illustrated
as aseptate with granulose content, and the width of ascomata
(140 μm) is too small to match the size of any accepted species
of Lentomitella. The type material of C. debaryana could not be
located. The holotype of E. conoidea deviates slightly in the
length of the asci from recently collected material from the Czech
Republic and Denmark; the asci are 57–66 × 6.5–7.5 μm,
although in the protologue of E. conoidea they were described as
longer, 62–72 × 6–8μmbyFeltgen (1903). Similar length of asci
of E. conoidea (ca. 70 μm) was reported also by Höhnel (1906b).
Lentomitella conoidea is represented by four strains in our
phylogeny. The ascospores are mostly 1–2-septate, the first
septum is formed in the middle or slightly above or below the
middle. Ascospores with three septa were observed only in the
collections from Denmark. The length of the neck and presence
of hairs covering the venter vary among collections.
Lentomitella conoidea is most similar to L. tenuirostris in
morphology of asci and ascospores, but it differs by slightly
shorter ascospores, slightly longer asci and larger ascomata. In
culture, the aerial mycelium of L. conoidea is woolly to cottony,
loose to woolly-floccose or almost cobwebby at the margin, while
in L. tenuirostris the aerial mycelium is cottony to felty resulting in
more compact aerial hyphae; the colony surface is paler,
because substrate mycelium and the brown hue in the agar do
R
EBLOV
AET AL.
20
not show through loose aerial hyphae as is the case of
L. conoidea. Moreover, the margin of the colony of L. conoidea
consists of densely branched hyphae of substrate mycelium,
while the margin of L. tenuirostris is formed of unbranched or
sparsely branched hyphae. Most specimens of L. conoidea were
collected on various hardwoods in deciduous forests in lowlands,
while specimens of L. tenuirostris were collected in the mountain
regions of the Czech Republic and France. For further com-
parison see Discussion and also comments of L. tenuirostris.
Lentomitella conoidea can be also compared to L. investita,
but the latter differs by shorter asci and ascospores that possess
the first-formed septum always in the middle and two additional
septa that form symmetrically.
Lentomitella crinigera (Cooke) R
eblov
a, Mycologia 98: 83.
2006. Fig. 11.
Basionym:Sphaeria crinigera Cooke, Grevillea 1: 156. 1873.
Synonyms:Ceratosphaeria crinigera (Cooke) Sacc. Syll. fung. 2:
227. 1883.
Fig. 10. Lentomitella conoidea.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D–F. Asci. G. Paraphyses. H. Ascal apex with apical annulus. I–K. Ascospores. L,
M. Colonies on MLA and PCA after 28 d. A, B, J, K from CBS 141370, C, E from M.R. 3135, F from LUX 043455, D, G, H, I, L, M from CBS 131660. Scale bars: A, B = 500 μm,
C, G = 20 μm, D–F=10μm, H –K=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 21
Fig. 11. Lentomitella crinigera.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D–F. Asci. G. Paraphyses. H, I. Colonies on MLA and PCA after 28 d. J. Ascal
apex with apical annulus. K, L. Ascospores. A–D, G–I, K from CBS 138678, E, J, L from M.R. 1671, F from M.R. 1526. Scale bars: A, B = 500 μm, C, G = 20 μm,
D–F=10μm, J–L=5μm.
R
EBLOV
AET AL.
22
Ceratostomella crinigera (Cooke) Cooke, Grevillea 17: 49. 1889.
Ceratostomella triseptata Petr., Annls mycol. 23: 135. 1925.
Sexual morph:Ascomata immersed with protruding necks or
partially erumpent becoming superficial with base immersed,
solitary, in rows or small groups. Venter 450–600 μm diam,
510–650 μm high, globose to subglobose, dark brown, with dark
brown, septate hairs 3.5–5μm wide. Neck central, 120–170 μm
wide, up to 1 mm long, cylindrical, upright, straight or slightly
flexuous, glabrous, tapering; apex sulcate. Ostiole periphysate.
Ascomatal wall fragile to leathery, 35–55(–65) μm thick, two-
layered; outer layer consisting of thick-walled, brown, poly-
hedral cells of textura angularis to textura prismatica with opaque
walls; cells tending to be darker towards the outside, becoming
flattened and paler towards the interior. Inner layer consisting of
several rows of thin-walled, hyaline, flattened cells. Paraphyses
abundant, longer than the asci, becoming disintegrated with age,
septate, constricted at the septa, hyaline, 6.5–10.5 μm wide,
tapering to 3.5–4.5 μm. Asci (66–)70–88(–90) × (7.5–)
8–9.5(–10) μm (mean ± SD = 78.7 ± 4.4 × 8.5 ± 0.6 μm), (62–)
68–76 μm (mean ± SD = 69.1 ± 4.4 μm) long in the sporiferous
part, truncate at the apex, cylindrical to clavate, with a short stipe;
with 8 obliquely uniseriate or biseriate ascospores; apical
annulus ca. 2.5–3μm wide, 1.5–2μm high. Ascospores (10–)
10.5–13 × 4–5.5 μm (mean ± SD = 11.5 ± 0.3 × 4.8 ± 0.3 μm),
ellipsoidal-fusiform, slightly inequilateral, hyaline, longitudinally
striate, 1–3-septate. Asexual morph: unknown.
Culture characteristics: Colonies on MLA 10–13 mm diam after
14 d (13–15 mm after 21 d, 15–17 after 28 d) at 25 °C, circular,
convex. Aerial mycelium abundant, woolly to cottony, margin
filamentous, colony surface pale brown to beige with a dark
brown marginal ring; reverse black. Colonies on PCA 4–5mm
diam after 14 d (5–7 mm after 21 d, 6 –8 mm after 28 d) at 25 °C,
circular, convex. Aerial mycelium abundant, cottony, loose to-
wards the margins, margin filamentous, colony surface beige to
pale brown, with a brown ring of submerged hyphae at the
margin; reverse black. Vegetative hyphae branched, septate,
medium to dark brown, smooth-walled. Margin of the colony
consisting of densely branched hyphae of substrate mycelium.
Sporulation not observed.
Specimens examined:Czech Republic, Northern Moravia, Podho
rí ‘Podhorn’
near Hranice, on decaying coniferous wood, Nov. 1913, F. Petrak (holotype of
Ceratostomella triseptata, W 18093). Southern Bohemia, Novohradsk
e hory Mts.,
Dobr
a voda, Hojn
a voda National nature monument, on decaying wood, 4 Oct.
2012, M. R
eblov
a M.R. 3742 (culture CBS 138678). Southern Bohemia,
Sumava
Mts. National park,
Zelezn
a Ruda, glacial cirque of the
Certovo jezero Lake
National nature reserve, on decaying wood of Abies alba, 28 Aug. 1997, M.
R
eblov
a, M.R. 997, M.R. 1034; ibid., on decaying wood of Picea abies 12 Aug.
1999, M.R. 1544, M.R. 1585; 21 Aug. 2000, M.R. 1672; ibid., glacial cirque of the
Cern
e jezero Lake National nature reserve, on decaying wood of Abies alba,13
Aug. 1999, M.R. 1526, M.R. 1546; ibid., Boubínský prales National nature
reserve, on decaying wood of Picea abies, 17 Aug. 1999, M.R. 1611; ibid.,
Pr
a
sily,
Zdanidla Mt., 24 Aug. 2000, on decaying wood of Picea abies, M.R.
1671.; ibid., Modrava, Pta
cí n
adr
z, on decaying wood of Picea abies, 14 Aug.
1999, M.R. 1457; ibid., Modrava, Modravsk
e slat
e, Pytl
acký roh, on decaying
wood of Picea abies, 14 Aug. 1999, M.R. 1652. Sweden, Fries'Scleromyceti
Sueciae No. 346, on decaying wood (UPS). UK, England, Norfolk, King's Lynn,
on decaying pine wood, C.B. Plowright (holotype of Sphaeria crinigera,K
84422).
Notes:Lentomitella crinigera is characterised by 1–3-septate,
ellipsoidal-fusiform ascospores, with all three septa developed
early in the ontogeny. Compared to other Lentomitella spp. the
ascomata of L. crinigera are larger, exceeding 500 μm diam, and
the neck is wider, 120–170 μm. It is probably the only member of
the genus which expresses a clear preference for coniferous
wood. The examination of the holotype of Ceratostomella tri-
septata revealed that this name is a synonym of L. crinigera.
Lentomitella crinigera is most similar to L. obscura,L. sulcata
and L. striatella in morphology and size of ascospores and asci.
Based on phylogenetic evidence, none of these three species is
closely related to L. crinigera and their distinction is supported at
the RNA structural level. However, in the absence of DNA
sequence data, morphological distinction of these species is
challenging. Both L. striatella and L. sulcata are known only from
New Zealand and were collected on decaying wood of Notho-
fagus sp.; the length of their asci is in the upper range typical of
L. crinigera. On the other hand, the asci of L. obscura are shorter
than those of L. sulcata and L. striatella and their length is in the
lower range of L. crinigera.Lentomitella obscura can be distin-
guished from L. crinigera in having slightly shorter ascospores
and by occurrence on deciduous wood; all three strains of
L. obscura originate from various deciduous hardwoods from
three localities in Ari
ege, southern France. Therefore, the host
determination is important to aid the identification of these
morphologically similar species.
The substrate of the specimen of L. crinigera sequenced in
this study (M.R. 3742, culture CBS 138678) has not been
determined. The wood is very rotten and was collected in a lo-
cality with stands of Fagus sylvatica,Picea abies and old fallen
trunks of Abies alba. Our attempts to isolate DNA from herbarium
material or cultivate L. crinigera from identified coniferous sub-
strates were not successful.
Lentomitella investita (Schw.) R
eblov
a, comb. nov. MycoBank
MB821766. Fig. 12.
Basionym:Sphaeria investita Schw., Trans. Amer. Phil. Soc. 2,
Vol. 4: 216. 1834.
Synonyms:Ceratostoma investitum (Schw.) Ellis & Everh., North
Amer. Pyrenom. p. 193. 1892.
Ceratostomella investita (Schw.) Starb€
ack, Bih. Kongl. Svenska
Vet.-Akad. Handl. 19(2): 26. 1894.
Amphitrichum investitum (Schw.) Kuntze, Revis. Gen. Pl. 3(2):
443. 1898.
Ceratostomella vestita Sacc. var. varvicensis Grove, J. Bot. 23:
131. 1885.
Ceratostomella maderensis Petr., Bot. Jahrb., Beiblt. 142: 98.
1929.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks becoming superficial with only base immersed,
solitary or in groups. Venter 300–350(–400) μm diam,
320–360 μm high, globose to subglobose, dark brown to black,
with sparse, brown to reddish-brown, septate hairs 3.5–4μm
wide at the lower part. Neck central, 90–110 μm wide, up to
800 μm long, cylindrical, upright, straight or slightly flexuous,
glabrous, tapering, apex sulcate. Ostiole periphysate. Ascomatal
wall fragile to leathery, 37–56 μm thick, two-layered; outer layer
consisting of thick-walled, brown, polyhedral cells of textura
angularis to textura prismatica with opaque walls; cells tending to
be darker towards the outside, becoming flattened and paler
towards the interior. Inner layer consisting of several rows of thin-
walled, hyaline, flattened cells. Paraphyses becoming dis-
integrated with age, septate, constricted at the septa, hyaline,
4.5–10 μm wide, tapering to ca. 3.5 μm. Asci (46–)
50–56(–65) × 6–7μm (mean ± SD = 54.3 ± 2.3 × 6.7 ± 0.3 μm),
45–52 μm (mean ± SD = 49.3 ± 2.3 μm) long in the sporiferous
part, truncate at the apex, cylindrical to clavate, with a short stipe;
SORDARIOMYCETES
www.studiesinmycology.org 23
with 8 obliquely uniseriate, partly overlapping ascospores; apical
annulus ca. 2.5–3μm wide, 1.5 μm high. Ascospores (7–)
7.5–9 × 3.5–4(–4.5) μm (mean ± SD = 8.1 ± 0.3 × 3.7 ±
0.3 μm), ellipsoidal or ellipsoidal-fusiform, slightly inequilateral,
hyaline, longitudinally striate, 1(–3)-septate, first-formed septum
in the middle, with two large drops. Asexual morph:A
phaeoisaria-like asexual morph was formed in vitro. Sporulation
sparse, widespread throughout the colony. Conidiophores mac-
ronematous, mononematous, arising terminally or laterally from
aerial hyphae, unbranched or branched apically, brown near the
base, subhyaline to hyaline towards the tip, 64–80 × 2–2.5 μm.
Conidiogenous cells terminal or intercalary, pale brown to sub-
hyaline at the bottom, hyaline in the conidiogenous part,
subcylindrical or slender flask-shaped, tapering toward the apex,
with a rachis 16–20 × 1.5–2μm at the tip; conidiogenesis
holoblastic-denticulate, conidial heads slimy, inconspicuous,
transparent. Conidia clavate to obovate, slightly apiculate at the
base, broadly rounded at the apical end, 4.5 –7×2
(mean ± SE = 5.2 ± 0.9 × 2) μm, hyaline, aseptate, smooth.
Specimens examined:Germany, Nassau, on decaying wood, in autumn 1894, L.
Fuckel (as Ceratostomella rhenana, Herb. Barbey-Boissier 603, G). Great
Britain, Sutton, Driffold Lane, on decaying wood, 25 Oct. 1884, W.B. Grove
(holotype of Ceratostomella vestita var. varvicensis, K 124145). Madeira,
Ribeiro frio, on decaying wood of Laurus novocanariensis, 3 Jun. 1926, A. Ade
(holotype of Ceratostomella maderensis, W 03981). New Zealand, South Island,
Southland, Southland Distr., Fiordland National park, Lake Monowai 40 km S of
Manapouri, Borland Nature Walk 300 m NW of Borland Lodge, on decaying wood
Fig. 12. Lentomitella investita.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D–G. Asci. H–J. Ascospores. K. Ascal apex with apical annulus. L, M. Co-
nidiophores. N. Conidia. A–E, H, I, K –N from PDD 110876, F, G, J from PH 01016198. Scale bars: A, B = 500 μm, C = 20 μm, D–G=10μm, H–N=5μm.
R
EBLOV
AET AL.
24
of Nothofagus sp., 9 Mar. 2005, M. R
eblov
a M.R. 2951/NZ 443 (PDD 110876);
ibid., West Coast, Buller Distr., Victoria Forest Park, Big River Inanganua track
14 km SE of Reefton, on decaying wood of Nothofagus sp., 6 Mar. 2003, M.
R
eblov
a M.R. 2721/NZ 222, M.R. 2726/NZ 227; ibid., Westland Distr., Arthur's
Pass National Park, Kelly Shelter 5 km W of Otira, Cockayane Nature Walk, on
decaying wood of Nothofagus sp., 16 Mar. 2003, M. R
eblov
a M.R. 2829/NZ 339.
USA, Pennsylvania, Northhampton County, Bethlehem, on decaying wood, L.D.
Schweinitz, S.F. 1621.476 (holotype of Sphaeria investita, PH 01016198).
Notes:Lentomitella investita is characterised by ellipsoidal,
mostly 1-septate ascospores with the septum positioned in the
middle, with two additional septa developed symmetrically but
rarely upon aging only in the collections from New Zealand. The
longitudinally striate walls of the ascospores are conspicuous
and individual ridges are often seen protruding at the poles. The
holotypes of Ceratostomella vestita var. varvicensis (Grove
1885) and Ceratostomella maderensis (Petrak 1929) were
examined. Based on the morphology of ascomata, asci and
ascospores they are considered conspecific with L. investita. The
holotype of C. maderensis deviates slightly in the length of the
asci from the holotype of L. investita and other recently collected
material; they are 52–63(–68) × 6–7.5 μm. Our specimens from
New Zealand fit well the description of L. investita, although the
ascospores in PDD 110876 are more tapering towards the ends.
This collection was isolated into axenic culture, which is, unfor-
tunately, no longer available. The fungus formed in vitro a
phaeoisaria-like asexual morph, which is consistent with asexual
morphs observed in L. sulcata and Lentomitella sp. The DNA
extraction from herbarium material was not successful.
Lentomitella investita,L. vestita and L. cirrhosa were formerly
treated as conspecific(von Arx 1952, R
eblov
a 2006). Based on
molecular evidence the two latter are accepted as separate
species in our study. Although no DNA sequence data of
L. investita are available, given the size of asci and ascospores,
the species is intermediate between L. vestita and L. cirrhosa and
therefore regarded as distinct. Lentomitella cirrhosa differs from
L. investita by longer and wider ascospores and longer asci. On
the other hand, L. vestita is well-distinguished from L. investita by
shorter, regularly 1-septate ascospores and shorter asci.
Lentomitella investita resembles L. conoidea in the size of
ascospores, but the latter differs by longer asci and mostly 1–2-
septate ascospores with the first-formed septum in the middle or
slightly above or below the middle.
Lentomitella magna R
eblov
a, sp. nov. MycoBank MB821767.
Fig. 13.
Etymology:Magnus (L.) meaning big, large, referring to the large
ascospores, which are the largest of the known Lentomitella
species.
Sexual morph:Ascomata immersed to partially erumpent
becoming superficial with only base immersed, solitary. Venter
300–390 μm diam, 310–380 μm high, globose to subglobose,
dark brown to black, with dark brown, septate hairs 3–3.5 μm
wide growing at the exposed sides. Neck central, 90–110 μm
wide, up to 2 000 μm long, cylindrical, upright or decumbent,
flexuous, glabrous, tapering, sometimes laterally flattened, apex
sulcate. Ostiole periphysate. Ascomatal wall fragile to leathery,
50–56 μm thick, two-layered; outer layer consisting of thick-
walled, brown, polyhedral cells of textura angularis to textura
prismatica with opaque walls; cells tending to be darker towards
the outside, becoming flattened and paler towards the interior.
Inner layer consisting of several rows of thin-walled, hyaline,
flattened cells. Paraphyses abundant, longer than the asci,
becoming disintegrated with age, septate, constricted at the
septa, hyaline, 5.5–7.5 μm wide, tapering to ca. 3 μm. Asci (89–)
93–102(–105) × (9.5–)10–12 μm (mean ± SD = 97.5 ± 4.7 ×
10.7 ± 1.4 μm), (75–)88–95 μm (mean ± SD = 88.1 ± 6.0 μm)
long in the sporiferous part, truncate at the apex, cylindrical
to clavate, with a short stipe; with 8 obliquely uniseriate or
partly biseriate ascospores; apical annulus 3.5 –4μm wide,
2μm high. Ascospores (12.5–)13–15 × 5.5–6.5 μm
(mean ± SD = 13.7 ± 0.9 × 6 ± 0.4 μm), ellipsoidal to ellipsoidal-
fusiform, inequilateral, hyaline, with longitudinal ridges that are
often discontinuous, 3-septate with four large drops. Asexual
morph: unknown.
Culture characteristics: Colonies on MLA 10–12 mm diam after
14 d (13–14 mm after 21 d, 15–16 after 28 d) at 25 °C, circular
to slightly irregular, mostly flat, raised at the centre. Aerial
mycelium sparse, cottony at the centre surrounded by a wide
zone of loose to almost cobwebby mycelium, margin filamen-
tous, colony surface beige in the centre with olive-brown ring of
submerged hyphae at the margin; reverse black. Colonies on
PCA 3–4 mm diam after 14 d (4–6 mm after 21 d, 5–7 mm after
28 d) at 25 °C, circular to slightly irregular, flat, raised at the
centre. Aerial mycelium sparse, cottony to woolly, cobwebby
towards the margin, margin filamentous, colony surface pale
brown with dark olive-brown ring of submerged hyphae at the
margin; reverse black. Vegetative hyphae branched, septate,
medium to dark brown, smooth-walled. Margin of the colony
consisting of densely branched hyphae of substrate mycelium.
Sporulation not observed.
Specimen examined:New Zealand, South Island, West Coast, Westland Distr.,
Westland Tai Poutini National park, Lake Matheson, 5 km SW from Fox Glacier,
on decaying wood, 13 Apr. 2005, M. R
eblov
a M.R. 2961/NZ 781 (holotype, PDD
110877, culture ex-type ICMP 18371).
Notes:Lentomitella magna is characterised by 3-septate,
ellipsoidal-fusiform, inequilateral ascospores and asci, which are
the largest of all Lentomitella species accepted in this study. It is
known only from a single collection from New Zealand. The
ascomata are mostly superficial with decumbent or upright necks
and horizontally lying venter covered with dark brown hairs. The
position of ascomata and necks on the substrate was influenced
by their growth under decaying, partly peeled off bark. The
longitudinal ridges in the ascospore wall are partly discontinuous
giving the ascospore wall a reticulate appearance.
Lentomitella obscura R
eblov
a, sp. nov. MycoBank MB821768.
Fig. 14.
Etymology:Obscure (L.) meaning indistinct, obscure, referring to
the morphology of ascospores.
Sexual morph:Ascomata immersed with protruding necks to
partially erumpent or becoming superficial, solitary or densely
aggregated. Venter 380–500 μm diam, 370 –510 μm high,
globose to subglobose, dark brown to black, with brown to
reddish-brown, septate hairs 3.5–4μm wide. Neck central,
120–150 μm wide, up to 1 500 μm long, cylindrical, upright or
partly decumbent, straight or slightly flexuous, glabrous, tapering,
apex sulcate. Ostiole periphysate. Ascomatal wall fragile to
leathery, 35–50 μm thick, two-layered; outer layer consisting of
thick-walled, brown, polyhedral cells of textura angularis to tex-
tura prismatica with opaque walls; cells tending to be darker
towards the outside, becoming flattened and paler towards the
interior. Inner layer consisting of several rows of thin-walled,
SORDARIOMYCETES
www.studiesinmycology.org 25
Fig. 13. Lentomitella magna.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E–H. Asci. I. Paraphyses. J, K. Ascospores. L. Ascal apex with apical annulus. M,
N. Colonies on MLA and PCA after 28 d. A–N from ICMP 18371. Scale bars: A–C = 500 μm, D, I = 20 μm, E–H=10μm, J –L=5μm.
R
EBLOV
AET AL.
26
Fig. 14. Lentomitella obscura.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E, F. Asci. G. Paraphyses. H. Ascal apex with apical annulus. I–K. Ascospores. L,
M. Colonies on MLA and PCA after 28 d. A–C, F, G, I from CBS 138735, D, E, H, J–M from CBS 138736. Scale bars: A–C = 500 μm, D, G = 20 μm, E, F = 10 μm,
H–K=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 27
hyaline, flattened cells. Paraphyses abundant, longer than the
asci, becoming disintegrated with age, septate, constricted at the
septa, hyaline, 5–8μm wide, tapering to ca. 3 μm. Asci (70–)
72–80(–82) × 7.5–8(–8.5) μm (mean ± SD = 76.2 ± 3.2 ×
8.1 ± 0.2 μm), 65–76 μm (mean ± SD = 71.8 ± 3.4 μm) long in
the sporiferous part, truncate at the apex, cylindrical to clavate,
with a short stipe; with 8 obliquely uniseriate or partly biseriate
ascospores; apical annulus ca. 3 μm wide, 1.5–2μm high. As-
cospores (10–)11–12 × 4–5(–5.5) μm (mean ± SD = 11.7
± 0.6 × 5 ± 0.4 μm), ellipsoidal-fusiform, hyaline, longitudinally
striate, 1–3-septate. Asexual morph: unknown.
Culture characteristics: Colonies on MLA 19–23 mm diam after
14 d (30–32 mm after 21 d, 37–42 after 28 d) at 25 °C, circular,
convex. Aerial mycelium abundant, woolly, loose towards the
margins, margin filamentous, colony surface pale brown-beige
with a brown ring of submerged hyphae at the margin; reverse
black. Colonies on PCA 7–10 mm diam after 14–28 d in CBS
138736; 10–13 mm after 14 d in CBS 138735 and CBS 137799
(20–21 mm after 21 d, 27–28 mm after 28 d) at 25 °C; circular to
irregular, convex. Aerial mycelium abundant, cottony to woolly,
loose towards the margins, margin filamentous, colony surface
beige-grey with olive-brown ring of submerged hyphae at the
margin; reverse black. Vegetative hyphae branched, septate,
medium to dark brown, smooth-walled. Margin of the colony
consisting of unbranched or sparsely branched hyphae of sub-
strate mycelium. Sporulation not observed.
Specimens examined:France, Ari
ege, Rimont, Las Muros, banks of the Peyrau
brook 440 m a.s.l., on decaying wood of Pinus sp., 30 Sep. 2013, M. R
eblov
a
M.R. 3707 (PRA-12736, culture CBS 138735); ibid., banks of La Maille brook,
550 m a.s.l., on decaying wood, 2 Oct. 2013, M. R
eblov
a M.R. 3784 (holotype,
PRA-12737, culture ex-type CBS 138736); ibid., along D18 1.5 km S of the
village, banks of Le Baup brook, on decaying wood of Quercus sp., 3 Oct. 2013,
M. R
eblov
a M.R. 3801 (culture CBS 137799).
Notes: The specimens of L. obscura were collected in three
different localities in the Ari
ege region in France. The ascomatal
necks vary greatly in length within one collection. They are
elongated up to 1 500 μm in more humid areas, especially in
cracks of wood, and much shorter (ca. 600 μm) when growing on
the surface of the wood, where they are more exposed to air and
sun. The hairs, which abundantly cover the venter, disappear
with age leaving the ascomata almost glabrous, but remnants of
hairs can be seen as a tomentum tightly attached to the base of
ascomata and wood.
Although all three strains of L. obscura form a strongly sup-
ported monophyletic clade and their DNA sequences are identical,
the morphological delimitation of this species was hindered by
different stages of their growth and maturation resulting in a var-
iable morphology of the ascospores. Two specimens, i.e. M.R.
3707 (culture CBS 138735) and M.R. 3801 (culture CBS 137799),
showed mostly ellipsoidal, 1–2-septate ascospores with the third
septum developing rarely and mostly in the shrinking ascospores.
Moreover, their ascospores measuring 9–10(–11) × 4 –4.5(–5)
μm and asci 65–72(–75) × 6.5 –7.5 μm were shorter than those of
the third strain, which is designated as holotype (PRA-12737, ex-
type strain CBS 138736). The holotype of L. obscura exhibits a
perfectly mature stage with ellipsoidal-fusiform ascospores that
are regularly 3-septate. In vitro, the colonies of all three strains
have identical appearance on MLA and PCA media except that the
ex-type strain grows slower on PCA (7–10 mm vs. 27–28 mm
after 28 d). Given these circumstances, it is extremely difficult to
identify the strains with shorter ascospores based on sexual
characters only; in such case at least ITS sequences should be
produced for further comparison.
In the phylogenetic tree (Fig. 1), L. obscura is placed with
Lentomitella sp., L. magna and L. sulcata in a highly supported
subclade; all these species having ellipsoidal-fusiform, 3-septate
ascospores. The latter three species all originate from New
Zealand. Lentomitella obscura is similar to L. sulcata in
morphology and size of ascospores, but the latter differs in
longer asci and it is delimited at the RNA structural level by a
unique CBC (Fig. 4), see Discussion.
Lentomitella striatella R
eblov
a, sp. nov. MycoBank
MB821769. Fig. 15.
Etymology: Diminutive of Striatus (L.) meaning striped, referring
to a fine linear marking in the longitudinally striate ascospores.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks, solitary or in groups. Venter 330–400 μm
diam, 310–410 μm high, globose to subglobose, dark brown to
black, with sparse, dark brown, septate hairs ca. 3.5 μm wide.
Neck central, 90–110 μm wide, 600 –1 500 μm long, cylindrical,
upright, straight, glabrous, tapering, apex sulcate. Ostiole
periphysate. Ascomatal wall fragile to leathery, 40–44 μm thick,
two-layered; outer layer consisting of thick-walled, brown, poly-
hedral cells of textura prismatica with opaque walls; cells tending
to be darker towards the outside, becoming flattened and paler
towards the interior. Inner layer consisting of several rows of thin-
walled, hyaline, flattened cells. Paraphyses abundant, longer
than the asci, becoming disintegrated with age, septate, con-
stricted at the septa, hyaline, 6.5–9.5 μm wide, tapering to ca.
4μm. Asci (75–)82–88 × 9–10.5 μm (mean ± SD = 85 ± 3.9
× 9.8 ± 0.6 μm), 68–78(–81) μm (mean ± SD = 75.8 ± 4.4 μm)
long in the sporiferous part, truncate at the apex, cylindrical to
clavate, with a short stipe; with 8 obliquely uniseriate or partly
biseriate ascospores; apical annulus 3–3.5 μm wide, 1.5–2μm
high. Ascospores 11.5–13(–14) × 5–5.5 μm (mean ± SD = 12
± 0.7 × 5.2 ± 0.3 μm), ellipsoidal, slightly inequilateral, hyaline,
longitudinally striate, 3-septate. Asexual morph: unknown.
Culture characteristics: Colonies on MLA 12–16 mm diam after
14 d (15–19 mm after 21 d, 18–21 after 28 d) at 25 °C, circular,
raised. Aerial mycelium abundant, cottony, loose towards the
margin, colony surface brown to dark olive-brown turning brown
towards the margin; reverse black. Colonies on PCA 11–12 mm
diam after 14 d (13–16 mm after 21 d, 14–18 mm after 28 d) at
25 °C, circular, raised. Aerial mycelium abundant, cottony to
woolly, loose towards the margin, colony surface brown-grey with
olive-brown ring of submerged hyphae at the margin; reverse
black. Vegetative hyphae branched, septate, medium to dark
brown, smooth-walled. Margin of the colony consisting of un-
branched or sparsely branched hyphae of substrate mycelium.
Sporulation not observed.
Specimens examined:New Zealand, South Island, West Coast, Buller Distr.,
Victoria Forest Park, Duffys Creek Track, 30 km SE from Reefton, on decaying
wood of Nothofagus sp., 7 Apr. 2005, M. R
eblov
a M.R. 2959/NZ 751 (holotype,
PDD 110878, culture ex-type ICMP 18369); ibid., Palmers Hut 18 km SW of
Springs Junction on unpaved road, Lake Christabel track, on decaying wood of
Nothofagus sp., 1 Mar. 2003, M. R
eblov
a M.R. 2694/NZ 195; ibid., Westland
Distr., Arthur's Pass National park, Arthur's Pass village, on decaying wood of
Nothofagus solandri, 17 Mar. 2003, M. R
eblov
a M.R. 2870/NZ 387.
Notes:Lentomitella striatella appears as sister to L. cirrhosa and
together they form a strongly supported monophyletic subclade
at the base of the Lentomitella clade. It differs from L. cirrhosa in
R
EBLOV
AET AL.
28
having longer and wider asci and longer, 3-septate ascospores.
The septa are formed early in ontogeny still within the asci, while
ascospores of L. cirrhosa are mostly 1-septate with two addi-
tional septa observed occasionally in released and shrinking
ascospores.
In morphology of ascospores, L. striatella resembles
L. crinigera,L. magna,L. obscura and L. sulcata.Lentomitella
crinigera and L. sulcata differ from L. striatella by shorter asco-
spores and slightly longer asci, and L. magna is well-distinguished
from L. striatella by longer ascospores and asci, while L. obscura
possesses much shorter asci. The ascospore wall of L. striatella is
longitudinally striate, but the ridges are shallow and appear less
conspicuous compared to other species.
Lentomitella sulcata R
eblov
a, sp. nov. MycoBank MB821770.
Fig. 16.
Etymology:Sulcatus (L.) meaning furrowed or grooved, referring
to the apex of the ascomatal neck, which has several deep
sulcations.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks, solitary. Venter 300–400 μm diam,
310–400 μm high, globose to subglobose, dark brown to black,
with sparse dark brown to reddish-brown, septate hairs 3–4μm
wide covering the lower part. Neck central, 100–120 μm wide, up
to 700 μm long, cylindrical, upright, straight or slightly flexuous,
glabrous, apex sulcate. Ostiole periphysate. Ascomatal wall
fragile to leathery, 30–45 μm thick, two-layered; outer layer
consisting of thick-walled, brown, polyhedral cells of textura
angularis to textura prismatica with opaque walls; cells tending to
be darker towards the outside, becoming flattened and paler
towards the interior. Inner layer consisting of several rows of thin-
Fig. 15. Lentomitella striatella.A. Ascomata. B. Longitudinal section of the ascomal wall. C, D. Asci. E. Paraphyses. F. Ascal apex with apical annulus. G, H. Ascospores. A
from M.R. 2694, B–H from ICMP 18369. Scale bars: A = 500 μm, B, E = 20 μm, C, D = 10 μm, F –H=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 29
R
EBLOV
AET AL.
30
walled, hyaline, flattened cells. Paraphyses longer than the asci,
becoming disintegrated with age, septate, constricted at the
septa, hyaline, 8–12.5 μm wide. Asci 79–90(–93) ×
8.5–9.5(–10) μm (mean ± SD = 86.4 ± 4.0 × 9.2 ± 0.4 μm),
72–84(–86) μm (mean ± SD = 80.3 ± 4.1 μm) long in the
sporiferous part, broadly rounded to truncate at the apex, cy-
lindrical to clavate, with a short stipe; with 8 obliquely uniseriate
ascospores; apical annulus ca. 3 μm wide, 2 μm high. Asco-
spores (10.5–)11–12(–13) × 5–5.5 μm (mean ± SD =
11.7 ± 0.5 × 5.3 ± 0.3 μm), ellipsoidal to ellipsoidal-fusiform,
slightly inequilateral, hyaline, longitudinally striate, 1–3-septate
with four large drops, when vital. Asexual morph:Co-
nidiophores in vitro macronematous, mononematous, arising
terminally or laterally from aerial hyphae, unbranched or rarely
branched apically, pale brown near the base, subhyaline to hy-
aline towards the tip, 24–45 × 2–2.5 μm. Conidiogenous cells
terminal or intercalary, hyaline, cylindrical, tapering toward the
apex, with a rachis 9–20(–25) × 2–2.5 μm at the tip bearing
2–10 hyaline denticles 0.5–1 × 0.5–1μm; conidiogenesis
holoblastic-denticulate, conidial heads slimy, inconspicuous,
transparent. Conidia ellipsoidal to globose, apiculate at the base,
4–6×2–2.5 μm (mean ± SE = 5.2 ± 0.2 × 2.2 ± 0.3 μm), hya-
line, aseptate, smooth.
Culture characteristics: Colonies on MLA 19–22 mm diam after
14 d (23–25 mm after 21 d, 25–26 after 28 d) at 25 °C, circular,
convex. Aerial mycelium abundant, cottony to woolly, margin
filamentous, colony surface beige-brown with olive-brown mar-
ginal ring of submerged hyphae; reverse black. Colonies on PCA
12–15 mm diam after 14 d (15–17 mm after 21 d, 18–19 mm
after 28 d) at 25 °C, circular, convex. Aerial mycelium abundant,
woolly, loose towards the margin, margin filamentous, colony
surface beige-grey with dark olive-brown marginal ring of sub-
merged hyphae; reverse black. Vegetative hyphae branched,
septate, medium to dark brown, smooth-walled. Margin of the
colony consisting of unbranched or sparsely branched hyphae of
substrate mycelium. Sporulation copious, widespread throughout
the colony.
Specimen examined:New Zealand, South Island, West Coast, Buller Distr.,
Victoria Forest park, Reefton, Lake Stream track, 30 km SE of Reefton, on
decaying wood of Nothofagus sp., 27 Feb. 2003, M. R
eblov
a M.R. 2659/NZ 145
(holotype, PDD 81435, culture ex-type ICMP 15124 = CBS 113655).
Notes: Although the specimen PDD 81435 was formerly treated
under L. crinigera (R
eblov
a 2006), it is introduced as a new
species, L. sulcata, in this study. Their distinction as two different
species, though morphologically difficult, is corroborated by
molecular and RNA structural data.
Lentomitella sulcata is nested together with L. magna,
L. obscura and Lentomitella sp. in a monophyletic clade. It is
characterised by 3-septate, longitudinally striate ascospores and
it forms simple, rarely branched conidiophores with globose to
ellipsoidal, apiculate conidia in vitro. The asexual morph is most
similar to Lentomitella sp. and also comparable to that produced
by L. investita.
Lentomitella tenuirostris R
eblov
a, sp. nov. MycoBank
MB821771. Fig. 17.
Etymology:Tenuis (L.) meaning thin, rostrum (L.) meaning beak,
referring to ascomata with a slender projecting neck.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks, becoming superficial, solitary, sometimes in
rows or small groups. Venter 300–410 μm diam, 300 –420 μm
high, globose to subglobose, black, upright sometimes posi-
tioned horizontally towards the substrate, with sparse, dark
brown, septate hairs 3–4μm wide growing from the bottom and
lower part, disappearing with age, venter later appears almost
glabrous. Neck central, 90–110 μm wide, up to 900 μm long,
cylindrical, upright, straight or slightly flexuous, tapering, apex
sulcate. Ostiole periphysate. Ascomatal wall fragile to leathery,
32–42 μm thick, two-layered; outer layer consisting of thick-
walled, brown, polyhedral cells of textura prismatica with opa-
que walls; cells tending to be darker towards the outside,
becoming flattened and paler towards the interior. Inner layer
consisting of several rows of thin-walled, hyaline, flattened cells.
Paraphyses abundant, longer than the asci, becoming dis-
integrated with age, septate, constricted at the septa, hyaline,
5–9μm wide, tapering to ca. 3.5 μm. Asci (58–)60–66(–69)
×7–7.5(–8) μm (mean ± SD = 62.9 ± 3.0 × 7.4 ± 0.5 μm),
52–62 μm (mean ± SD = 57.7 ± 3.2 μm) long in the sporiferous
part, truncate at the apex, cylindrical, with a short stipe; with 8
uniseriate partially overlapping or obliquely uniseriate asco-
spores; apical annulus ca. 2.5 μm wide, 1.5 μm high. Ascospores
(8.5–)9–10.5 × 4–4.5 μm (mean ± SD = 9.8 ± 0.6 × 4.3 ± 0.3
μm), ellipsoidal to suboblong, slightly inequilateral, sometimes
slightly curved, hyaline, longitudinally striate, 1–3-septate,
mostly 1–2-septate, the first-formed septum in the middle or
slightly above or below the middle, formation of the third septum
delayed, visible in old ascospores. Asexual morph: unknown.
Culture characteristics: Colonies on MLA 10–13 mm diam after
14 d (12–14 mm after 21 d, 19–20 mm after 28 d) at 25 °C,
circular, convex to pulvinate. Aerial mycelium abundant, cottony
to felty, loose to floccose towards the margin, margin filamen-
tous, colony surface beige to brown-grey with outer olivaceous
ring and darker olive-brown ring of submerged hyphae at the
margin; reverse black. Colonies on PCA 7–10 mm diam after
14 d (9–10 mm after 21 d, 10–12 after 28 d) at 25 °C, circular,
convex. Aerial mycelium abundant, cottony to felty, floccose to-
wards the margin, margin filamentous, colony surface beige-grey
with dark olive-brown ring consisting of submerged hyphae at the
margin; reverse black. Margin of the colony consisting of un-
branched or sparsely branched hyphae of substrate mycelium.
Sporulation not observed.
Specimens examined:Czech Republic, Southern Bohemia, Novohradsk
e hory
Mts., Poho
rí na
Sumav
e, Myslivna Mt., on decaying wood of Sorbus aucuparia,6
Oct. 2012, M. R
eblov
a M.R. 3771 (holotype, PRA-12738, culture ex-type CBS
138734); ibid., Dobr
a voda, Hojn
a voda National nature monument, on decaying
wood of Fagus sylvatica, 4 Oct. 2012, M.R. 3735; ibid., 28 Sep. 2014, M.R. 3859.
Sumava Mts. National park,
Zelezn
a Ruda, glacial cirque of the
Cern
e jezero lake
National nature reserve, on decaying wood of Picea abies, 22 Oct. 1998, M.R.
858; ibid., Boubínský prales National nature reserve, on decaying wood of Fagus
sylvatica, 14 Aug. 1999, M.R. 1533; ibid., Mt. Sp
aleni
st
e near Sto
zec, on
decaying wood of Fraxinus excelsior, 16 Aug. 1999, M.R. 1545; ibid., Povyd
rí
National nature reserve,
Ce
nkova Pila, on decaying wood of Ulmus glabra,27
Aug. 2000, M.R. 1677. France, Ari
ege, Rimont, Las Muros, banks of the Peyrau
Fig. 16. Lentomitella sulcata.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E–G. Asci. H. Paraphyses. I, J. Ascospores. K. Ascal apex with apical annulus. L,
M. Conidiophores. N. Conidia. O, P. Colonies on MLA and PCA after 28 d. A–P from ICMP 15124. Scale bars: A –C = 500 μm, D, H = 20 μm, E –G=10μm, I –N=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 31
brook 440 m a.s.l., on decaying wood, 30 Sep. 2013, M. R
eblov
a M.R. 3715
(culture CBS 141371), M.R. 3712, M.R. 3714.
Notes:Lentomitella tenuirostris and L. conoidea form a pair of
closely related and morphologically highly similar species. There
are only a few subtle differences in ascus and ascospore
characters and the colony appearance in vitro, which make their
correct identification challenging in the absence of molecular
data. Lentomitella tenuirostris differs from L.conoidea by slightly
shorter asci, slightly longer ascospores and generally smaller
ascomata clothed by sparse dark brown to black hairs, but the
venter soon becomes glabrous. The development of the third
septum in ascospores of L. tenuirostris was observed in
shrinking ascospores still within the asci only in specimen M.R.
3712 collected in the same locality and on the same day as
specimen M.R. 3715, whose culture CBS 141371 was
sequenced in our study. Only two strains of L. tenuirostris were
isolated in axenic culture. Several other specimens from France
and the Czech Republic fit well the description of this species.
Lentomitella tenuirostris can be distinguished from the
morphologically similar L. investita by shorter asci and shorter,
mostly 1-septate ascospores of the latter. Lentomitella uni-
pretoriae (Marincowitz et al. 2008) has similar ascospore size,
but it differs from L. tenuirostris by regularly 3-septate asco-
spores and longer asci.
Fig. 17. Lentomitella tenuirostris.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E, F. Asci. G. Ascal apex with apical annulus. H. Paraphyses. I–K. Ascospores.
L, M. Colonies on MLA and PCA after 28 d. A, B, E, G from CBS 141371, C, D, H, J–M from CBS 138734, F, I from M.R. 3712. Scale bars: A–C = 500 μm, D, H = 20 μm, E,
F=10μm, G, I –K=5μm.
R
EBLOV
AET AL.
32
Lentomitella unipretoriae M.J. Wingf. et al., CBS Biodiversity
Ser. 7: 60. 2008.
Notes: For description and illustration see Marincowitz et al.
(2008). The authors emphasized the evanescent nature of pa-
raphyses as a unique character of this species, however, this is a
character common to all members of Lentomitella.Lentomitella
unipretoriae originates from South Africa where it was collected
on a senescent flower head of Protea lepidocarpodendron.Itis
most similar to L. crinigera,L. magna,L. striatella and L. sulcata
in 3-septate, ellipsoidal ascospores, but differs from them by
smaller ascospores whose length does not exceed 11 μm, (8–)
9–10.5(–11) × 4 –5μmfide Marincowitz et al. (2008). No DNA
sequence data nor a living culture of this species are available.
Lentomitella vestita (Sacc.) Höhn., Annls mycol. 3: 548. 1906.
Fig. 18.
Basionym:Ceratostomella vestita Sacc., Michelia 1: 370. 1878.
Synonyms:Cerastomis vestita (Sacc.) Clem., Gen. Fungi p. 259.
1931.
Endoxyla vestita (Sacc.) Munk, Bot. Tidsskr. 61: 64. 1965.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks or becoming superficial, solitary or in small
groups. Venter 300–380 μm diam, 310 –400 μm high, globose to
subglobose, dark brown to black, with dark brown, septate hairs
4‒4.5 μm wide covering the lower part. Neck central, 90 –110 μm
wide, up to 600 μm long, cylindrical, upright, straight, glabrous,
apex sulcate. Ostiole periphysate. Ascomatal wall fragile to
leathery, 33–40 μm thick, two-layered; outer layer consisting of
thick-walled, brown, polyhedral cells of textura angularis to tex-
tura prismatica with opaque walls; cells tending to be darker
towards the outside, becoming flattened and paler towards the
interior. Inner layer consisting of several rows of thin-walled,
Fig. 18. Lentomitella vestita.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D. Asci. E. Paraphyses. F. Ascal apex with apical annulus. G–K. Ascospores. A –K
from PRA-12739. Scale bars: A, B = 500 μm, C, E = 20 μm, D = 10 μm, F –K=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 33
hyaline, flattened cells. Paraphyses abundant, longer than the
asci, becoming disintegrated with age, septate, constricted at the
septa, hyaline, 6–10 μm wide, tapering to 3.5–4μm. Asci (40–)
42–51(–55) × (5.5–)6–7μm (mean ± SD = 47 ± 3.9 × 6.5 ± 0.3
μm), 36–45(–48) μm (mean ± SD = 41.2 ± 3.1 μm) long in the
sporiferous part, truncate at the apex, cylindrical to clavate, with
a short stipe; with 8 obliquely uniseriate ascospores; apical
annulus 2–2.5 μm wide, 1.5 μm high. Ascospores (5.5–)
6–7×3–3.5 μm (mean ± SD = 6.7 ± 0.4 × 3.3 ± 0.2 μm),
ellipsoidal, broadly rounded at both ends but sometimes slightly
tapering, hyaline, longitudinally striate, 1-septate. Asexual
morph: unknown.
Specimens examined:Czech Republic, Central Bohemia, Lys
a nad Labem,
By
si
cky, Hrb
a
ckovy tůn
e National nature reserve, on decaying wood of Quercus
sp., 11 Nov. 2012, M. R
eblov
a M.R. 3677 (PRA-12739). Denmark, Fyn, Snarup
Mose SW of Kværndrup, on inner side of bark of Betula sp., 1 Apr. 1999, J.
Vesterholt JV99-015 (C 45296); ibid., Sjælland, Charlottenlund skov, on decaying
wood, 10 Nov. 1964, A. Munk (C); ibid., Dyrehaven, on decaying wood, 26 Nov.
1964, A. Munk (C); ibid., on decaying wood of Fagus sylvatica, 2 Dec. 1963, A.
Munk (C). Germany, Oestrich forest, on decaying wood, autumn, L. Fuckel
(Fungi Rhen. Exs. No. 1804, G); ibid., on decaying wood of Fagus sylvatica,L.
Fuckel (as Ceratostomella rostrata, Herbarium Barbey-Boissier, G). Italy,on
decaying wood, Sep. 1878. P.A. Saccardo (holotype of Ceratostomella vestita,
PAD).
Notes:Lentomitella vestita is well-distinguished from other
Lentomitella species by the shortest ascospores and asci in the
genus and the 1-septate ascospores that do not develop addi-
tional septa. The neck in the holotype does not appear sulcate,
i.e. the tip of the neck is without longitudinal ridges and instead it
appears only slightly roughened. However, the material is old
and the ridges could have disappeared upon aging and some of
the tips were broken. All other collections of L. vestita that were
examined in this study have ascomata with necks distinctly
sulcate at the tip, which is a character common to all Lentomitella
species.
Lentomitella vestita resembles L. investita in 1-celled ellip-
soidal ascospores, but the latter differs in having longer asci and
longer and wider ascospores that occasionally develop two
additional septa upon aging.
Despite our numerous attempts we did not obtain L. vestita in
culture. Therefore, the DNA was extracted from herbarium ma-
terial of our collection PRA-12739 that matches the holotype.
Some ascospores in this collection seem to taper slightly at the
poles, which is caused to a certain extent by protrusion of the fine
longitudinal ridges, a character we also observed in L. investita.
Lentomitella sp. Fig. 19.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks, solitary. Venter 390–480 μm diam,
370–500 μm high, globose to subglobose, dark brown to black,
with sparse brown, septate hairs 3.5–4μm wide. Neck central,
110 –130 μm wide, up to 700 μm long, cylindrical, upright,
straight or slightly flexuous, glabrous, tapering, apex sulcate.
Ostiole periphysate. Ascomatal wall fragile to leathery,
35–43 μm thick, two-layered; outer layer consisting of thick-
walled, brown, polyhedral cells of textura angularis to textura
prismatica with opaque walls; cells tending to be darker towards
the outside, becoming flattened and paler towards the interior.
Inner layer consisting of several rows of thin-walled, hyaline,
flattened cells. Paraphyses abundant, becoming disintegrated
with age, septate, constricted at the septa, hyaline, 4.5–8μm
wide. Asci (61–)63–70(–75) × (7.5)8–9.5 μm (mean ± SD =
67.1 ± 4.5 × 8.7 ± 0.7 μm), (55–)60–63 μm (mean ± SD =
61.1 ± 4.7 μm) long in the sporiferous part, truncate at the apex,
cylindrical to clavate, with a short stipe; with 8 obliquely uni-
seriate or partly biseriate ascospores; apical annulus ca. 3 μm
wide, 1.5–2μm high. Ascospores 9.5–11 × 4.5–5μm
(mean ± SD = 10.5 ± 0.5 × 4.8 ± 0.3 μm), ellipsoidal-fusiform,
inequilateral, hyaline, longitudinally striate, 1–3-septate.
Asexual morph: Sporulation in vitro sparse, widespread
throughout the colony. Conidiophores macronematous, mono-
nematous, unbranched or rarely branched apically, arising
terminally or laterally from aerial hyphae, pale brown, subhyaline
towards the tip, 40–45 × 2–2.5 μm. Conidiogenous cells terminal
or intercalary, pale brown or subhyaline, cylindrical, slightly
tapering toward the apex, with a rachis 15–25 × 2(–2.5) μm, at
the tip bearing numerous hyaline denticles 0.5–1μm wide,
0.5–1μm long, conidiogenesis holoblastic-denticulate, conidial
heads slimy, inconspicuous, transparent. Conidia ellipsoidal to
globose, apiculate at the base, 2–3(–4) × 2–3 (mean ± SE =
3.1 ± 0.9 × 2.6 ± 0.4) μm, hyaline, aseptate, smooth.
Specimen examined:New Zealand, South Island, Southland Distr., Catlins
Coastal Rain Forest park, Haldane, Kiwi Walk along the Waipohatu stream, at the
end of Waipohatu Road, 11 km W of Waikawa, 14 Mar. 2005, on decaying wood,
M.R. 2953/NZ 500.
Notes: This collection from New Zealand is grouped in a
monophyletic clade consisting of this and the three species
L. magna,L. obscura and L. sulcata; all are characterised by 3-
septate, ellipsoidal-fusiform, slightly inequilateral ascospores. It
is most similar to L. sulcata, which also originates from New
Zealand, in morphology of ascospores and conidia but differs in
shorter ascospores and asci. For comparison of this clade at the
RNA structural level see Discussion. The specimen M.R. 2953
was scarce and contained only a few ascomata; because no
material is left after isolation and examination, and the culture is
no longer available, this specimen is labelled Lentomitella sp. for
the time being and its morphological description, illustration and
DNA sequences are published in this study.
Spadicoides S. Hughes, Canad. J. Bot. 36: 805. 1958; emend.
R
eblov
a & A.N. Mill.
Synonyms:Xenospadicoides Hern.-Restr. et al., Stud. Mycol. 86:
92. 2017.
Pseudodiplococcium Hern.-Restr. et al., Stud. Mycol. 86: 92.
2017.
Sexual morph:Ascomata perithecial, non-stromatic, immersed to
partially erumpent with protruding necks, solitary, in short rows or
grouped. Venter globose or subglobose, often pinched laterally
upon drying, dark brown to black, clothed with septate dark hairs;
surface sometimes covered by a bright waxy layer staining the
surrounding substrate. Neck central, upright, sometimes slightly
decumbent, glabrous, the projecting part dark brown, light
fulvous to subhyaline, cylindrical, apex smooth or with several
sulcations. Ostiole periphysate. Ascomatal wall fragile to
leathery, two-layered. Paraphyses becoming disintegrated
apically, anastomosing, septate. Asci cylindrical to clavate, with a
short stipe, 8-spored, apex with a distinct, non-amyloid apical
annulus. Ascospores ellipsoidal to ovoid, often inequilateral,
aseptate or with a middle septum usually developed upon aging,
hyaline, smooth-walled or delicately verrucose. Asexual morph:
Colonies dark, effuse, stroma absent. Conidiophores macro-
nematous, mononematous, unbranched, occasionally with
branches, erect or ascending, straight or flexuous, septate,
brown, paler towards the apex. The upper three-quarters or less
R
EBLOV
AET AL.
34
of the conidiophores are usually conidiogenous. Conidiogenous
cells terminal or intercalary in the upper part of the conidiophore,
cylindrical, with numerous small pores in the wall, conidia formed
singly or in chains at the apex and laterally, conidiogenesis tretic.
The number of pores per conidiogenous cell usually 5–15
causing the conidia to entirely envelope the conidiophore in a dry
mass. Conidia oblong, ellipsoidal or obovoid, aseptate or
several-septate, brown, borne singly or occasionally in chains at
the apex and laterally in the position of pores and secede readily.
Synasexual morph: A selenosporella-like is sometimes formed
in vitro and in vivo.Conidiophores macronematous or semi-
macronematous, mononematous, branched or unbranched,
often reduced to conidiogenous cells. Conidiogenous cells
discrete, terminal or intercalary, subcylindrical to flask-shaped,
polyblastic, sympodially proliferating with a short terminal
rachis, often arising from vegetative hyphae or directly from
conidia, sometimes reduced to a few denticles. Conidia clavate,
oblong or narrow fusiform, hyaline, aseptate, smooth-walled.
Type species:Spadicoides bina (Corda) S. Hughes
Notes: Based on a revision of the holotypes of the two Cera-
tostomella species, their novel DNA sequence data and cultures,
and nucLSU sequence of the ex-type strain of Pseudodiplo-
coccium ibericum (Hern
andez-Restrepo et al. 2017), three new
combinations are proposed in Spadicoides, namely S. fuscolutea
with Lentomitella tomentosa and S. grovei as synonyms,
S. hyalostoma and S. iberica.
The sexual-asexual relationship of Spadicoides was experi-
mentally proven in our study for S. bina,S. fuscolutea and
S. hyalostoma.Spadicoides atra and S. iberica remain asexual.
Fig. 19. Lentomitella sp. A. Longitudinal section of the ascomal wall. B. Paraphyses. C. Ascal apex with apical annulus. D. Ascospores. E, F. Asci. G–J. Conidiophores. K.
Conidia. A–K from M.R. 2953. Scale bars: A, B = 20 μm, C, D, G–K=5μm, E, F = 10 μm.
SORDARIOMYCETES
www.studiesinmycology.org 35
R
EBLOV
AET AL.
36
A selenosporella-like synasexual morph has been encountered
for S. bina and S. fuscolutea in vitro (see Discussion). Given
these results, the description of Spadicoides is expanded to
include sexual and asexual characters.
The Spadicoides sexual morphs, especially S. bina and
S. fuscolutea, are strongly reminiscent of Lentomitella in
morphology of ascomata with a sulcate neck and venter clothed
by dark interwoven hairs and cylindrical short-stipitate asci with a
distinct apical annulus. However, Lentomitella differs from Spa-
dicoides by 1–3-septate, longitudinally striate vs. aseptate or
rarely 1-septate, smooth-walled or verrucose ascospores.
The main mode of conidiogenesis of Spadicoides is tretic,
rarely accompanied by a holoblastic-denticulate conidiogenesis
of a selenosporella-like synasexual morph. A key to holomorphic
Spadicoides is provided below. Keys to asexual morphs of
Spadicoides were published in Ellis (1963), Wang (1976),
Holubov
a-Jechov
a (1982), Goh & Hyde (1996) and Ma et al.
(2016).
Key to sexually reproducing Spadicoides species
Spadicoides bina (Corda) S. Hughes [as ‘binum’], Canad. J.
Bot. 36: 806. 1958. Fig. 20.
Basionym:Helminthosporium binum Corda [as ‘Helmisporium’],
in Zobel, Icon. Fung. 6: 9. 1854.
For full synonymy see Hughes (1958).
Sexual morph:Ascomata non-stromatic, immersed to partially
erumpent with protruding necks, solitary, in rows or small groups.
Venter 420–510 μm diam, 390–500 μm high, globose to sub-
globose, slightly pinched laterally upon drying, upright or lying
horizontally, dark brown to black, with dark brown, septate hairs
3–3.5 μm wide covering the lower part. Neck central,
100–120 μm wide, up to 800 μm long, rostrate, glabrous, upright,
apex sulcate becoming deeply roughened with age. Ostiole
periphysate. Ascomatal wall fragile to leathery, 45–55( –63) μm,
two-layered; outer layer consisting of brown, polyhedral cells of
textura prismatica to textura angularis with opaque walls; cells
tending to be darker towards the outside, and becoming flattened
and paler towards the interior. Inner layer consisting of several
rows of thin-walled, hyaline, flattened cells. Paraphyses abun-
dant, longer than the asci, becoming slightly disintegrated at the
apex, septate, constricted at the septa, hyaline, 4–5μm wide,
tapering to ca. 2.5 μm. Asci 60–67(–73) × (7.5–)8–9μm
(mean ± SD = 65.1 ± 1.5 × 8.4 ± 0.4 μm), 52–58 μm
(mean ± SD = 54.8 ± 2.6 μm) long in the sporiferous part, trun-
cate at the apex, cylindrical to clavate, with a short stipe, as-
cospores overlapping uniseriate or partly biseriate; with 8
uniseriate to partially biseriate ascospores; apical annulus ca.
2.5 μm wide, 1.5 μm high. Ascospores (7.5–)
8–9.5 × 3.5–4.5 μm (mean ± SD = 8.8 ± 0.5 × 3.9 ± 0.4 μm),
ellipsoidal to ovoid, aseptate, rarely with a cytoplasmic band in
the middle or a middle septum developed in old and shrinking
ascospores, hyaline, with one or two large drops, smooth-walled.
Asexual morph: Remnants of conidiophores and conidia identical
to those developed in vitro were observed in the intimate
juxtaposition to ascomata; conidia aseptate 6.5–7 × 3.5 μm, 1-
septate 7.5–8 × 3.5 μm. Conidiophores in vitro macro-
nematous, 30–75 μm long, 2.5–3μm wide, unbranched or with
short branches, erect, straight or flexuous, septate, brown, paler
towards the apex, smooth-walled. Conidiogenous cells inte-
grated, terminal or intercalary in the upper part of the conidio-
phore, 2.5–3μm wide, cylindrical, polytretic with numerous small
pores in the wall. Conidia mostly aseptate, 5–6×3–3.5 μm
(mean ± SD = 5 ± 0.5 × 3.4 ± 0.3 μm), pale to brown, ellipsoidal,
rounded at the apical end, with a slightly apiculate basal scar,
thick-walled, formed singly through pores or in short chains, at
maturity becoming 1-septate, 7–9.5 × 3.5–4μm
(mean ± SD = 7.9 ± 1.1 × 3.8 ± 0.2 μm), pale brown to dark
brown, ellipsoidal to oblong, with the middle septum obscured by
a black band, slightly constricted at the septum, smooth-walled.
Synasexual morph: A selenosporella-like was formed on MLA
and PCA in 28 d at 25 °C. Conidiophores semimacronematous,
branched or unbranched, pale brown to subhyaline, often
reduced to conidiogenous cells. Conidiogenous cells discrete or
integrated, terminal, intercalary, often arising directly from dark
brown, aseptate conidia, hyaline, subcylindrical or narrowly flask-
shaped, 6.5–11.5 μm long, ca. 2 μm wide, tapering to 1–1.2 μm,
apically slightly swollen or elongated, polyblastic, sympodially
proliferating with a short terminal rachis. Conidia
3.5–4.5 × 1–1.5 μm (mean ± SD = 3.8 ± 0.3 × 1.3 ± 0.2 μm),
clavate, oblong, slightly curved, hyaline, aseptate, smooth-
walled.
Culture characteristics: Colonies on MLA 14–15 mm diam after
14 d (20–23 mm after 21 d, 34–35 mm after 28 d) at 25 °C,
circular, raised. Aerial mycelium abundant, cottony to woolly,
margins filamentous, colony surface beige-brown with a thin
brown margin of submerged hyphae, the margin disappears as
colony ages and is overgrown by aerial hyphae; reverse dark
brown. Colonies on PCA 15–16 mm diam after 14 d (19–20 mm
after 21 d, 22–23 mm after 28 d) at 25 °C, circular, flat. Aerial
mycelium abundant, cottony to felty, margins filamentous, colony
surface beige with a brown margin of submerged hyphae, margin
disappears as colony ages and is overgrown by aerial hyphae;
reverse dark brown. Vegetative hyphae branched, septate, pale
brown, 1.5–2.5 μm wide, smooth-walled. Sporulation in 21 d at
the margin of the colony.
Specimen examined:Czech Republic, Southern Moravia, Valtice, Rendez-vous
National nature monument, on decaying wood of Quercus cerris, 17 Nov. 2012,
M. R
eblov
a M.R. 3686 (PRA-13420, culture CBS 137794).
Notes:Spadicoides bina is most similar to S. fuscolutea in
morphology of ascomata, asci and ascospores, but the latter
differs in presence of orange-red pigment on the outer ascomatal
wall layer, larger ascospores, asci and obovate or clavate, 3-
septate conidia.
The conidia of S. bina are 0–1-septate, occasionally a second
septum can develop (Ellis 1963). Hughes (1973a) examined the
type, “cotype”(syntype) and other authentic material and
Fig. 20. Spadicoides bina.A, B. Ascomata. C. Longitudinal section of the ascomal wall. D. Asci. E. Paraphyses. F. Ascospores. G. Shrinking ascospores with a middle septum
(see arrow). H. Conidia. I, K. Selenosporella-like synasexual morph. J, L–N. Conidiophores with conidia (arrows indicate conidia in short chains). O, P. Colonies on MLA and
PCA after 28 d. A–G from PRA-13420, H–P from CBS 137794. Scale bars: A, B = 500 μm. C = 20 μm, D, E, I, J, L –N=10μm, F, G, H, K = 5 μm.
1. Ascospores delicately verrucolose ………………………….... S. hyalostoma
1. Ascospores smooth-walled …………………………………………………….2
2. Ascospores (7.5–)8–9.5 × 3.5–4.5 μm……………………………… S. bina
2. Ascospores (13–)14–16(–17) × (5 –)5.5 –6(–7) μm………... S. fuscolutea
SORDARIOMYCETES
www.studiesinmycology.org 37
R
EBLOV
AET AL.
38
concluded that the number of septa in conidia can vary in
different collections. Ellis (1963) illustrated 1-septate conidia of
S. bina as usually oblong to oval, non-constricted or slightly
constricted with a conspicuous reddish-brown band obscuring
the middle septum. Hughes (1973a) noted that 1-septate conidia
are “generally waisted”at the middle septum. The 1-septate
conidia observed in vivo and those formed in vitro in the strain
CBS 137794 were always slightly constricted at the middle
septum. The formation of the septum was delayed, conidia were
mostly aseptate. Spadicoides bina resembles S. canadensis
(Hughes 1973b) in 1-septate, brown, oblong to ellipsoidal con-
idia, but the latter species differs by somewhat wider conidia
(5.2–6.3 μmfide Hughes 1973b) and a septum formed slightly
below the middle of the conidium. Spadicoides bina occurs on
decaying wood and bark of various deciduous and coniferous
trees (Ellis 1963, Hughes 1973a, Holubov
a-Jechov
a 1982).
Spadicoides fuscolutea (Rehm) R
eblov
a, comb. nov. Myco-
Bank MB821772. Fig. 21.
Basionym:Ceratostomella fuscolutea Rehm, Annls mycol. 6:
320. 1908.
Synonyms:Lentomitella tomentosa R
eblov
a & J. Fourn.,
Mycologia 98: 86. 2006.
Spadicoides grovei M.B. Ellis, Mycol. Pap. 93: 12. 1963.
Diplococcium grovei (M.B. Ellis) R.C. Sinclair et al., Trans. Br.
mycol. Soc. 85: 736. 1986.
Sexual morph:Ascomata non-stromatic, immersed with pro-
truding necks or becoming superficial, solitary, in short rows or
groups. Venter 350–500 μm diam, 400 –550 μm high, globose to
subglobose, pinched laterally upon drying, dark brown to black,
with septate, pale brown to reddish-brown hairs ca. 3–5μm
wide; surface of the venter covered by an orange to orange-red
waxy layer up to 13 μm thick sometimes disappearing with age;
granules of the same pigment also attached to the surface of
hairs and also staining the surrounding substrate. Neck central,
100–150(–180) μm wide, 500–700 μm long, cylindrical, upright,
tapering, apex with 3–5 deep sulcations. Ostiole periphysate.
Ascomatal wall fragile to leathery, 45–55 μm, two-layered; outer
layer consisting of brown, thick-walled, polyhedral cells of textura
prismatica to textura angularis with opaque walls. Inner layer
consisting of several rows of thin-walled, hyaline, flattened cells.
Paraphyses becoming disintegrated apically, anastomosing,
septate, constricted at the septa, hyaline, 4.5–6μm wide,
tapering to 2.5–3μm. Asci (75–)80–102(–110) × 9.5–11 μm
(mean ± SD = 89.1 ± 4.5 × 10.6 ± 1.1 μm), (65–)70–85(–90)
μm (mean ± SD = 73.9 ± 4.2 μm) long in the sporiferous part,
broadly rounded or truncate at the apex, cylindrical to clavate,
with a short stipe; with 8 obliquely uniseriate or biseriate asco-
spores in the upper sporiferous part; apical annulus 3–3.5 μm
wide, 2–2.5 μm high. Ascospores (13–)14–16(–17) × (5–)
5.5–6(–7) μm (mean ± SD = 14.5 ± 0.4 × 5.8 ± 0.3 μm), ellip-
soidal to ovoid, sometimes inequilateral, aseptate, rarely with a
middle septum, hyaline, smooth-walled. Asexual morph:Co-
nidiophores in vitro macronematous, 45–100 μm long, 3–4μm
wide, unbranched, rarely with short branches, erect, straight or
flexuous, septate, dark brown, smooth-walled. Conidiogenous
cells integrated, intercalary, terminal, positioned in the upper two-
thirds of the conidiophore, ca. 3.5(–4) μm wide, cylindrical, with
1–3 pores in the wall visible after the secession of conidia.
Conidia 16.5–20(–21) × (7.5–)8–10(–11) μm (mean ± SD
= 18.8 ± 1.3 × 8.8 ± 1.0 μm), formed singly through the pores,
obovate or clavate, rounded at the apical end, conico-truncate at
the base, brown to dark brown, 3-septate, often only with two
septa developed; septa thick and dark brown due to the band of
colour on the wall. Synasexual morph: A selenosporella-like was
formed on MLA in 8 wk at 25 °C. Conidiophores macronematous
or semimacronematous, branched, often reduced to con-
idiogenous cells, pale brown to hyaline, 10–55 μm long,
1.5–2μm wide. Conidiogenous cells integrated, terminal or
intercalary, subhyaline to hyaline, subcylindrical, 10–15 μm long,
ca. 2.5 μm wide near the base, tapering to 1.5 μm, polyblastic,
sympodially proliferating with a short terminal rachis. Conidia
4–5.5 × 1.5(–2) μm (mean ± SD = 4.6 ± 0.5 × 1.4 ± 0.2 μm),
clavate or oblong, slightly curved, hyaline, aseptate, smooth-
walled.
Culture characteristics: Colonies on MLA 8–10 mm diam after
14 d (16–17 mm after 21 d, 18–20 mm after 28 d) at 25 °C,
circular, convex. Aerial mycelium abundant, woolly, margins
filamentous, colony surface beige grey, brown olivaceous to-
wards the margin; reverse black to dark brown. Colonies on PCA
4–5 mm diam after 14 d (6–7 mm after 21 d, 13 –14 mm after
28 d) at 25 °C, circular, convex. Aerial mycelium abundant,
woolly, margins filamentous, colony surface brown-grey, brown-
olivaceous towards the margin; reverse black. Sporulation in
21 d at the margins of the colony.
Specimens examined:Czech Republic, Southern Bohemia, Novohradsk
e hory
Mts., Poho
rí na
Sumav
e, Myslivna Mt., on decaying wood of Fagus sylvatica,6
Oct. 2012, M. R
eblov
a M.R. 3776 (culture CBS 141263); ibid., Horní Stropnice,
Hojn
a Voda National nature monument, on decaying wood of Fagus sylvatica,4
Oct. 2012, M. R
eblov
a M.R. 3744 (culture CBS 141262); ibid., 13 Oct. 2013, M.R.
3813 (associated with Spadicoides hyalostoma). Denmark,Sjæland, Dyrehaven,
on decaying wood of a stump, 3 Oct. 1964, A. Munk (C); ibid., 20 Mar. 1965, A.
Munk (C). Silkeborg, Spring area near Almindsø, on decaying wood of Alnus
glutinosa, 4 Sep. 1953, A. Munk (C). France, Finist
ere, Plohars, For^
et de
Carno€
et, Pont Douar, 40 m a.s.l., on decaying wood of Fagus sylvatica, 26 Oct.
2002, J. Fournier J.F. 02196, associated with the asexual morph (holotype of
Lentomitella tomentosa, PRM 902274). Germany, Swabia, Allg€
au, Hochgrat Mt.,
on decaying wood of Fagus sylvatica, 1881, M. Britzelmayer [holotype of
Ceratostomella fuscolutea, F11132 (S)].
Notes:Spadicoides fuscolutea is distinguished from other
members of the genus by ascomata, which are covered by a thin
layer of orange-red pigment that also stains the surrounding
wood and 3-septate, brown to dark brown, obovate or clavate
conidia, which are truncate at the base and arise singly or in
short chains from polytretic conidiogenous cells. The bright
pigment dissolves in KOH.
The asexual morph was originally described by Ellis (1963) as
Spadicoides grovei from decaying wood of Fagus sylvatica and
other trees, while the sexual morph was introduced as Cera-
tostomella fuscolutea (Rehm 1908) and Lentomitella tomentosa
(R
eblov
a 2006), both from wood of F. sylvatica. The link between
the two morphs was experimentally proven by ascospore
isolation of two specimens M.R. 3744 (culture CBS 141262) and
M.R. 3776 (culture CBS 141263) collected on beech wood in the
Czech Republic. Both collections fit well into the species concept
Fig. 21. Spadicoides fuscolutea.A, B. Ascomata. C, D. Asci. E, F. Acospores. G. Longitudinal section of the ascomal wall. H. Paraphyses. I, J. Selenosporella-like synasexual
morph. K–M. Conidiophores with conidia. N, O. Colonies on MLA and PCA after 28 d. A–C, G from PRM 902274, D–F, H, I, J, N, O from CBS 141263, K–M from CBS
141262. Scale bars: A, B = 500 μm, C–F, H –M=10μm, G = 20 μm.
SORDARIOMYCETES
www.studiesinmycology.org 39
of C. fuscolutea. Identical conidia and remnants of conidiophores
were also observed in the holotype of L. tomentosa (PRM
902274) and in M.R. 3776 around ascomata on the wood or
trapped among the ascomatal hairs. Based on the evidence from
a taxonomic revision of the holotypes of C. fuscolutea and
L. tomentosa and cultivation data, C. fuscolutea is transferred to
Spadicoides and S. grovei and L. tomentosa are accepted as its
synonyms.
The development of the orange-red layer covering the venter
varies. For example, in the holotype of L. tomentosa (R
eblov
a
2006), this layer is conspicuous, while in specimens M.R.
3774 and M.R. 3776 the orange layer almost disappeared
probably as a result of aging, but the orange pigment staining the
surrounding wood is still visible. In the holotype of Ceratosto-
mella fuscolutea (F11132, S), the orange-pigmented wood sur-
rounding the ascomata is also prominent.
The asexual morph resembles S. xylogena in 3-septate,
brown conidia, but the latter taxon differs by longer, oval to
broadly ellipsoidal conidia with narrower black bands at the septa
(Ellis 1963, Hughes 1973c). Based on DNA sequence data,
S. xylogena is related to the Pleosporales (Shenoy et al. 2010).
Spadicoides constricta,S. klotzschii, and S. obovata resemble
S. fuscolutea in clavate, obovate or sometimes also ellipsoidal
conidia; S. constricta differs from S. fuscolutea by longer conidia
constricted at the septa (Wang & Sutton 1982), while S. klotzschii
and S. obovata differ by conidia that are 2-septate, shorter and
narrower (Hughes 1973d, e, Holubov
a-Jechov
a 1982).
Spadicoides hyalostoma (Munk) R
eblov
a, comb. nov. Myco-
Bank MB821773. Fig. 22.
Basionym:Endoxyla hyalostoma Munk, Bot. Tidsskr. 61: 62.
1965.
Synonym:Ceratostomella hyalostoma (Munk) Unter., Mycologia
85: 307. 1993.
Sexual morph:Ascomata non-stromatic, immersed to partially
erumpent with protruding necks, solitary or grouped. Venter
270–350 μm diam, 280–380 μm high, subglobose, often laterally
pinched upon drying, dark brown, glabrous with brown, septate
hyphae at the base. Neck central, 90–130 μm wide,
130–170 μm at the widest part, slightly narrower at the base,
250–800 μm long, cylindrical, upright, straight or slightly flex-
uous, the projecting part subhyaline to light fulvous with darker
zones. Ostiole periphysate. Ascomatal wall leathery, 35–42 μm,
two-layered; outer layer consisting of brown, polyhedral cells of
textura prismatica with opaque walls. Inner layer consisting of
several rows of thin-walled, hyaline, flattened cells. Cells in the
neck of textura porrecta with thick-walled clavate cells on the
surface. Paraphyses abundant, persistent, becoming dis-
integrated at the apex, septate, constricted at the septa, hyaline,
3.5–5μm wide, tapering to ca. 2.5 μm. Asci (36–)
40–45(–48) × (5–)5.5–7(–8) μm (mean ± SD = 43.5 ± 2.9 ×
6.1 ± 0.4 μm), truncate at the apex, cylindrical to clavate, non-
stipitate or with a short narrowly rounded stipe; with 8
obliquely uniseriate or partially biseriate ascospores in the lower
sporiferous part; apical annulus ca. 2 μm wide, 1.5–2μm high.
Ascospores (5.5–)6–7×3–3.5 μm
(mean ± SD = 6.1 ± 0.4 × 3.1 ± 0.2 μm), ellipsoidal to ovoid,
0–1-septate, with one or two large drops, hyaline, delicately
verruculose. Asexual morph:Conidiophores in vitro macro-
nematous, 30–130 μm long, 2.5–3μm wide, unbranched or with
short branches, upright, septate, brown, subhyaline or pale
brown when young, dark brown at maturity, smooth-walled.
Conidiogenous cells integrated, terminal, intercalary, positioned
in the upper two thirds of the conidiophore, ca. 2.5 μm wide,
cylindrical, polytretic with numerous pores dispersed per cell.
Conidia 4.5–6 × 3.5–4(–4.5) μm (mean ± SD = 5.4 ± 0.5
× 3.7 ± 0.4 μm), formed singly through the pores, subglobose to
ellipsoidal, sometimes inequilateral, tapering towards base and
truncate, aseptate, hyaline becoming dark brown.
Culture characteristics: Colonies on MLA 16–17 mm diam after
14 d (29–30 mm after 21 d, 40–41 mm after 28 d) at 25 °C,
circular, raised. Aerial mycelium abundant, cottony to woolly,
margins filamentous, colony surface beige with a brown margin
of submerged hyphae; reverse dark brown. Colonies on PCA
15–18 mm diam after 14 d (24–26 mm after 21 d, 27–28 mm
after 28 d) at 25 °C, circular, raised. Aerial mycelium abundant,
cottony to woolly, margins filamentous, colony surface beige-
grey, with a brown-olivaceous margin of submerged mycelium;
reverse dark brown. Vegetative hyphae branched, septate,
smooth-walled, 2–3.5 μm wide, hyaline; hyphae on which the
conidiophores arise brown. Periodically arranged clusters of cells
bearing short branches are present on submerged hyphae at the
colony margin. Sporulation observed after 2 mo at the margins of
the colony.
Specimens examined:Czech Republic, Central Bohemia, K
rivokl
atsko Pro-
tected Landscape Area, Karlova Ves, Vysoký Tok Nature Reserve, on decaying
wood of Quercus sp., 29 Sep. 2012, M.R. 3662 (culture CBS 137793). Southern
Bohemia, Novohradsk
e hory Mts., Horní Stropnice, Hojn
a Voda National nature
monument, on decaying wood of Fagus sylvatica, 13 Oct. 2013, M. R
eblov
a M.R.
3813, associated with S. fuscolutea (culture CBS 138688). Northern Moravia,
Podho
rí ‘Podhorn’near Hranice, on decaying wood, 7 Aug. 1923, F. Petrak (as
Ceratostomella subpilosa, Fl. Bohem. Morav. Exs. No. 1809, PRM 481212).
Southern Moravia, Lednice, Nejdek old Slavic settlement, on decaying wood of
Fraxinus excelsior, 27 Oct. 2014, M. R
eblov
a M.R. 3866 (culture CBS 139771);
ibid., B
reclav, obora Soutok near Lan
zhot, on decaying wood, 23 Oct. 2004, M.
R
eblov
aM.R. 2597. Denmark, Sjaelland, Ermelunden forest near Copenhagen,
on decaying wood, 15 Dec. 1963, A. Munk (holotype of Endoxyla hyalostoma,
collection no. 14, C); ibid., Høje Sandbjerg, decaying wood, 18 Dec. 1964, A.
Munk (C); ibid., Dyrehaven, on decaying wood, 3 Nov. 1964, A. Munk (C); ibid.,
Lellinge, decaying wood, 23 May 1964, A. Munk (C); ibid., Bernstorffsparken, on
decaying wood, 23 Mar. 1965, A. Munk (C); ibid., Boserup, on decaying wood of
Ulmus sp., soc. Capronia pilosella. 1 Apr. 1963, A. Munk (C); ibid., Rude Skov, on
decaying wood of Alnus sp., 4 Feb. 1964, A. Munk (C); ibid., on decaying wood of
Fagus sylvatica, 17 Nov. 1963, A. Munk (C); ibid., forest W of Frederikssun, on
decaying wood, soc. Ruzenia spermoides, A. Munk (C). Italy, Lazio, province
Viterbo, Caprarola, Mt. Venere, on decaying wood of Fagus sylvatica, 2 Apr.
2011, M. R
eblov
a M.R. 3610 (culture CBS 131268).
Notes:Spadicoides hyalostoma can be distinguished from other
species of the genus by subhyaline to light fulvous necks with
darker zones and verruculose, aseptate ascospores. A
selenosporella-like synasexual morph has not been observed
in vitro or on the natural substrate in any examined collection.
The asexual morph of S. hyalostoma strongly resembles
S. atra in the morphology of conidia. Although the size of conidia
of both species overlap, i.e. conidia of S. atra 4–6.5 × 3 –4μmfide
Ellis (1963), 3.7–6.3 × 2.5–4.6 μmfide Hughes (1973f) and
4–6.5 (7.5) × 2.5 –3.5 (4.5) μmfide Holubov
a-Jechov
a (1982), the
latter species is distinguished by conidia rounded at both ends.
Spadicoides atra is represented in our phylogeny by the strain
CBS 489.77 (Czech Republic, L
any, L
ansk
a obora game park, on
decaying wood of Quercus petraea, 12 Jun. 1976, V. Holubov
a-
Jechov
a) and it is shown basal to other Spadicoides species.
Spadicoides hyalostoma is also highly similar to S. subsphaerica,
but the latter species differs in having shorter, subglobose,
R
EBLOV
AET AL.
40
Fig. 22. Spadicoides hyalostoma.A–D. Ascomata. E. Asci. F. Paraphyses. G. Ascospores. H. Conidia. I–K. Conidiophores. L, M. Colonies on MLA and PCA after 28 d. A–D
from CBS 137793, E, G from CBS 139771, H–M from CBS 138688. Scale bars: A–D = 250 μm, E, F, I–K=10μm, G, H = 5 μm.
SORDARIOMYCETES
www.studiesinmycology.org 41
globose or broadly ellipsoidal conidia rounded at both ends (Li
2010). It can be also compared with several other Spadicoides
species with aseptate, brown conidia, i.e. S. arengae,S. cuneata,
S.macrocontinua, and S. sphaerosperma. These taxa differ from
S. hyalostoma by larger, differently shaped conidia.
Spadicoides iberica (Hern.-Restr. et al.)R
eblov
a & A.N. Mill.,
comb. nov. MycoBank MB823341
Basionym:Pseudodiplococcium ibericum Hern.-Restr. et al.,
Stud. Mycol. 86: 92. 2017.
Notes: For description and illustration see Hern
andez-Restrepo
et al. (2012,2017). This species produces branched co-
nidiophores and (0–)1-septate conidia in long often branched
chains.
Torrentispora K.D. Hyde et al., Mycol. Res. 104: 1399. 2000;
emend. Shearer & F.R. Barbosa, Mycologia 105: 338. 2013.
Synonyms:Pseudoannulatascus Z.L. Luo et al., Phytotaxa 239:
179. 2015.
Fusoidispora Vijaykr. et al., Sydowia 57: 272. 2005.
Sexual morph:Ascomata perithecial, non-stromatic, immersed,
partially erumpent becoming superficial with only bases
immersed, scattered or grouped, varying in position from upright
to nearly horizontal. Venter globose, subglobose to conical,
glabrous or sparsely clothed with hairs. Neck rostrate or cylin-
drical, without sulcations at the apex, dark brown, glabrous or
hairy, upright or lying horizontally on the host. Ostiole periph-
ysate. Ascomatal wall fragile, two-layered, with layers of cylin-
drical cells in surface view. Paraphyses becoming partially
disintegrated, tapering, septate. Asci unitunicate, cylindrical,
stipitate, 8-spored, apex with a non-amyloid, massive, refractive
apical annulus. Ascospores ellipsoidal, ellipsoidal-fusiform or
fusiform, or rarely cymbiform to cylindrical, often flattened on one
side, sometimes slightly curved, hyaline, aseptate or with several
transverse septa at maturity, thick-walled, smooth-walled or with
afibrillar sheath. Asexual morph: unknown.
Type species:Torrentispora fibrosa K.D. Hyde et al.
Notes:Torrentispora comprises eight species from freshwater
and terrestrial environments. It is characterised by immersed to
partially erumpent or almost superficial, mostly glabrous asco-
mata with a rostrate or cylindrical neck, disintegrating paraphy-
ses, hyaline, ellipsoidal to fusiform to elongate-fusiform
ascospores and a distinct apical annulus. Barbosa et al. (2013)
emended the generic description by including taxa with glabrous
or hairy ascomata, asci with simple or bipartite apical rings and
ascospores with or without a gelatinous sheath that become
septate at maturity. Four previously described species are
accepted, i.e. T. crassiparietis,T. fibrosa,T. fusiformis and
T. pilosa (Hyde et al. 2000, Fryar & Hyde 2004, Barbosa et al.
2013). Based on novel DNA sequences and morphology,
T. calembola and T. novae-zelandiae are described as new to
science and three new combinations in Torrentispora are pro-
posed below for Ceratostomella dubia,Fusoidispora aquatica
and Pseudoannulatascus biatriisporus. The genera Fusoidispora
and Pseudoannulatascus are synonymised with Torrentispora.
Torrentispora can be distinguished from other members of the
Xenospadicoidales in having ascomatal necks that are never
sulcate, a massive apical annulus and hyaline, thick-walled
ascospores (except in T. aquatica, see Discussion). The delayed
development of septa in ascospores observed in T. biatriispora,
T. crassiparietis and T. pilosa (Barbosa et al. 2013) is also typical
of the closely related Calyptosphaeria,Lentomitella and Spadi-
coides. A relatively large range in the ascospore length of Tor-
rentispora spp. causes frequent overlapping in the size of
ascospores of individual species. The proportion between the
length of the sporiferous part and stipe of the ascus may vary
slightly regarding the position and spacing of ascospores.
The asexual morph of Torrentispora is unknown. Two species,
T. fibrosa and T. novae-zelandiae were obtained in axenic cul-
ture, but only sterile dematiaceous mycelium is formed.
Key to species of Torrentispora
Torrentispora aquatica (Vijaykr. et al.)R
eblov
a & A.N. Mill.,
comb. nov. MycoBank MB821774.
Basionym:Fusoidispora aquatica Vijaykr. et al., Sydowia 57:
272. 2005.
Notes: For description and illustration see Vijaykrishna et al.
(2005). Based on the partial nucLSU sequence data of the holo-
type of Fusoidispora aquatica, this species grouped in Torrentis-
pora and a new combination is proposed. Torrentispora aquatica
was originally collected on wood submerged in fresh water in Hong
Kong, China. The main feature distinguishing T. aquatica from
other members of this genus are thin-walled, elongate-fusiform to
cymbiform, septate ascospores with globose mucilaginous pads at
1. Ascospores longer than 40 μm……………………………………..……... 2
1. Ascospores shorter than 40 μm…………………………...……………..... 3
2. Ascospores elongate-fusiform with slightly swollen ends enclosed in a
thin, irregular mucilaginous sheath, 40–58 × 8–10 μm…………………
………………………………………………………………….. T. biatriispora
2. Ascospores cymbiform to fusiform or cylindrical, with globose
mucilaginous pads at both ends, 42–50 × 4–6um………..…T. aquatica
3. Ascospores longer than 20 μm…………………………………………… 4
3. Ascospore length range from below 20 μm………...…………………….. 8
4. Ascospores 20–30 μm long ……………………………………………… 5
4. Ascospores longer than 30 μm…………………………………………… 7
5. Ascomatal neck glabrous ………………………………………………… 6
5. Ascomatal neck covered by short hairs, ascospores 21–30 × 7–8μm
…………………………………………………..…………………….T. pilosa
6. Neck rostrate, ascospores (19–)20–25(–27.5) × (6.5–)7–8μm………
………………………………………………………………………… T. dubia
6. Neck cylindrical, ascospores 22–25(–27) × 6.5–8(–8.5) μm…………
…….……………………………………………………………... T. calembola
7. Ascospores 32–48 × 8–14 μm, wall 2–3μm thick at sides, 3–4μmat
the ends ……………………..……………………………… T. crassiparietis
7. Ascospores 24–32.5 × 6 –9μm, thinner-walled …………..…T. fusiformis
8. Ascospores 13.5–19.5 × 5 –7μm………………………..……… T. fibrosa
8. Ascospores (17.5–)18–25(–26) × (6 –)7–8.5 μm….T. novae-zelandiae
R
EBLOV
AET AL.
42
both ends. The pads disappear soon after ascospores are
released from the asci (Vijaykrishna et al. 2005).
Torrentispora biatriispora (K.D. Hyde) R
eblov
a & A.N. Mill.,
comb. nov. MycoBank MB821775.
Basionym:Annulatascus biatriisporus K.D. Hyde, Nova Hedw.
61: 120 (1995)
Synonym:Pseudoannulatascus biatriisporus (K.D. Hyde) Z.L.
Luo et al., Phytotaxa 239: 179. 2015.
Notes: For description and illustration see Hyde (1995) and
Barbosa et al. (2013). Based on DNA sequence data of a
freshwater specimen of P. biatriisporus from Costa Rica, this
species was nested in the Torrentispora clade and a new com-
bination is proposed. Torrentispora biatriispora is distinguished
from other species of the genus by long fusiform ascospores
surrounded by thin irregular mucilage mostly at the poles and
weakly swollen ends. It was reported from submerged wood from
the tropics in the southern and northern hemispheres including
Australia (Hyde 1995), China (Tsui et al. 2002), Costa Rica
(Barbosa et al. 2013) and the Seychelles (Hyde & Goh 1998).
Torrentispora calembola R
eblov
a & A.N. Mill., sp. nov.
MycoBank MB821776. Fig. 23A–I.
Etymology:Cal- from kal
os (Gr.) meaning beautiful,
embolon
(Gr.) meaning the beak of a ship, referring to the long necks that
give the fungus a decorative look.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks becoming superficial, solitary or grouped, venter
300–420 μm diam, 310–450 μm high, subglobose to conical,
upright, dark brown to black, with brown, septate hairs 2–3μm
wide, sparsely covering the sides. Neck central, 100–140 μm
wide, up to 700 μm long, cylindrical, upright, flexuous, tapering,
apex roughened without sulcations. Ostiole periphysate. Asco-
matal wall fragile to leathery, 38–58 μm thick, two-layered; outer
layer consisting of thick-walled, brown, polyhedral cells of textura
prismatica to textura angularis with opaque walls; cells tending to
be darker towards the outside, becoming flattened and paler to-
wards the interior. Inner layer consisting of several rows of thin-
walled, hyaline, flattened cells. Paraphyses abundant, longer
than the asci, becoming disintegrated with age, septate, slightly
constricted at the septa, hyaline, 5–6.5 μm wide, tapering to
2.5–3μm. Asci (195–)207–236 × (9–)10–11.5 μm (mean ± SD =
216.4 ± 10.2 × 10.7 ± 0.7 μm), 162–191(–196) μm (mean ± SD
= 171.6 ± 11.1 μm) long in the sporiferous part, truncate at the
apex, cylindrical, stipitate; with 8 uniseriate ascospores; apical
annulus 5–5.5 μm wide, 4–5μmhigh.Ascospores 22–25(–27) ×
6.5–8( –8.5) μm (mean ± SD = 23.2 ± 1.3 × 7.1 ± 0.5 μm), ellip-
soidal to fusiform, flattened on one side, hyaline, aseptate,
smooth- and thick-walled (0.8–0.9 μm), usually with a large drop
when fresh. Asexual morph: unknown.
Specimens examined:Czech Republic, Southern Bohemia,
Sumava Mts. Na-
tional park,
Zelezn
a Ruda, glacial cirque of the
Certovo jezero lake, on decaying
wood of Fagus sylvatica, 23 Oct. 1996, M. R
eblov
a M.R. 903. Southern Moravia,
B
reclav distr., Valtice, U t
rí Gr
acií, on decaying wood of Quercus sp., 15 Nov.
2012, M. R
eblov
a M.R. 3679; ibid., Valtice, Rendez-vous National nature mon-
ument, on decaying wood of Quercus cerris, 28 Nov. 2013, M. R
eblov
a M.R.
3843. France, Ari
ege, Monts
egur, shores of the Le Lesset stream along D9 road,
880–890 m a.s.l., on decaying wood, 1 Oct. 2013, M. R
eblov
a M.R. 3726
(holotype, PRA-12744); ibid., Rimont, La Maille brook, ca. 550 m a.s.l., on
submerged wood of Fraxinus excelsior staining deep green, 13 Mar. 2014,
incubated in moist chamber until 28 Apr. 2014, J. Fournier J.F. 14027
(PRA-12745).
Notes: The five specimens of T. calembola from terrestrial and
freshwater habitats in the Czech Republic and France fit well
within the concept of the genus and are introduced as a new
species supported by molecular DNA data. Torrentispora
calembola closely resembles T. fusiformis and T. pilosa in
morphology and size of ascospores, but it can be distinguished
from T. pilosa in that the latter possesses shorter and narrower
asci and hairy ascomatal necks. The main features dis-
tinguishing T. fusiformis from T. calembola are smaller ascomata
and longer ascospores exceeding 30 μm. Torrentispora fusi-
formis is known only from a freshwater habitat in Brunei.
Because the ascospores of T. calembola did not germinate
in vitro, the DNA was extracted directly from the holotype.
Torrentispora crassiparietis Fryar & K.D. Hyde, Cryptog.
Mycol. 25: 255. 2004.
Notes: For description and illustration see Fryar & Hyde (2004)
and Barbosa et al. (2013). This species is known from sub-
merged wood in freshwater habitats from Brazil, Brunei and Costa
Rica. Torrentispora crassiparietis can be distinguished from other
members of the genus by having ascospores with considerably
thicker walls, 2–3μm thick at sides, 3–4μm at the ends fide
Barbosa et al. (2013). These authors also enlarged its description
based on material from Brazil and Costa Rica including hairy
ascomata and larger asci and ascospores that stain blue in
aqueous nigrosine and become 2–3-septate at maturity.
Torrentispora dubia (Sacc.) R
eblov
a & A.N. Mill., comb. nov.
MycoBank MB821777. Fig. 23J–S.
Basionym:Melanomma dubium Sacc., Fungi Veneti novi vel
critici. Series III. Michelia 1: 449. Fungi Italici Autographice
Delineati. Fasc. 5–8. fig. 299. 1878.
Synonyms:Zigno€
ella dubia (Sacc.) Sacc., Michelia 1: 346. 1878.
Ceratostomella dubia (Sacc.) Sacc., Syll. fung. 1: 410. 1882.
Amphitrichum dubium (Sacc.) Kuntze, Revis. gen. pl. 3(2): 443.
1898.
Sexual morph:Ascomata partially erumpent becoming superfi-
cial with only base immersed, solitary or grouped. Venter
300–370(–420) μm diam, 300–350(–450) μm high, sub-
globose to conical, upright or lying slightly horizontally on the
host, dark brown to black, glabrous or with brown, septate hairs
2–3μm diam, sparsely covering the bottom and exposed sides.
Neck central, 90 –110 μm wide, up to 500 μm long, conical to
rostrate, upright, apex without sulcations. Ostiole periphysate.
Ascomatal wall fragile to leathery, 37 –43 μm thick, two-layered;
outer layer consisting of thick-walled, brown, polyhedral cells of
textura prismatica to textura angularis with opaque walls; cells
tending to be darker towards the outside, becoming flattened
and paler towards the interior. Inner layer consisting of several
rows of thin-walled, hyaline, flattened cells. Paraphyses sparse,
longer than the asci, becoming disintegrated with age, septate,
slightly constricted at the septa, hyaline, 8–7μm wide, tapering
to 3–3.5 μm. Asci 180–240 × (9–)9.5–10.5 μm(mean±SD=
209.2 ± 5.9 × 9.8 ± 0.5 μm), 150–182(–192) μm(mean±SD
=175.3±6.0μm) long in the sporiferous part, truncate at the
apex, cylindrical, stipitate; with 8 uniseriate ascospores; apical
annulus 4.5–5.5 μm wide, 3.5–4.5 μmhigh.Ascospores (19–)
20–25(–27.5) × (6.5–)7 –8μm (mean±SD = 22.9±1.3
×7.6±0.5μm), ellipsoidal to fusiform, flattened on one side,
hyaline, aseptate, smooth- and thick-walled (0.6 μm), filled with
one large or numerous small drops. Asexual morph: unknown.
SORDARIOMYCETES
www.studiesinmycology.org 43
R
EBLOV
AET AL.
44
Specimens examined:Czech Republic, Southern Bohemia,
Sumava Mts. Na-
tional park, Pr
a
sily, Mt.
Zdanidla, on decaying wood of Fagus sylvatica, 24 Aug.
2000, M. R
eblov
a M.R. 1681; ibid., Sto
zec, Oslí vrch Mt., on decaying wood of
Sorbus aucuparia, 16 Oct. 2010, M. R
eblov
a M.R. 2985 (PRA-12744). Germany,
Eberbach, on decaying wood of Fagus sylvatica, Herb. Barbey-Boissier No. 620
(G). Italy, Treviso, Cansiglio, on decaying wood of Fagus sylvatica, Oct. 1874,
P.A. Saccardo (holotype of Melanomma dubium, PAD).
Notes: A revision of the type material of Melanomma dubium
(Saccardo 1878b), although in poor condition and containing only
a few perithecia on a piece of wood of Fagus sylvatica, revealed
a fungus that matches well the description of Torrentispora.A
specimen from Barbey-Boissier herbarium No. 620 (G) and both
collections from the Czech Republic correspond most closely to
M. dubia. The transfer of M. dubia to Torrentispora is supported
by novel DNA sequences of the Bohemian specimen PRA-
12744. All examined collections occur on strongly decayed
wood of deciduous trees with a preference for F. sylvatica.
Torrentispora dubia is easily distinguished by ascomata with a
conical rostrate neck vs. cylindrical neck in other members of
Torrentispora. It is most similar to T. calembola in morphology of
the ascospores, but it differs in shorter and narrower asci and
anatomy of the neck. Both species are known from terrestrial
habitats.
Torrentispora fibrosa K.D. Hyde et al., Mycol. Res. 104: 1399.
2000. Fig. 24.
Sexual morph:Ascomata immersed to partially erumpent with
protruding necks, solitary or grouped. Venter 300–340 μm diam,
310–360 μm high, subglobose, upright or lying horizontally on
the host, dark brown to black, with brown, septate hairs
2.5–4μm wide, sparsely covering the exposed sides and bot-
tom. Neck central, 90–110 μm wide, up to 600 μm long, cylin-
drical, upright or decumbent, apex without sulcations. Ostiole
periphysate. Ascomatal wall fragile to leathery, 32–45 μm thick,
two-layered; outer layer consisting of thick-walled, brown, poly-
hedral cells of textura prismatica to textura angularis with opaque
walls; cells tending to be darker towards the outside, becoming
flattened and paler towards the interior. Inner layer consisting of
several rows of thin-walled, hyaline, flattened cells. Paraphyses
abundant, longer than the asci, becoming disintegrated with age,
septate, slightly constricted at the septa, hyaline, 4.5–9μm wide,
tapering to ca. 3 μm. Asci (148–)152–186(–204) × 7.5–9×6–9
μm (mean ± SD = 168.3 ± 14.7 × 8.3 ± 0.5 μm), (107–)
116 –140(–162) μm (mean ± SD = 128.8 ± 11.8 μm) long in the
sporiferous part, truncate at the apex, cylindrical, stipitate, with 8
uniseriate, ascospores; apical annulus 4–4.5 μm wide,
2.5–3μm high. Ascospores (14.5–)15–18(–19) × 5.5–7μm
(mean ± SD = 15.8 ± 1.1 × 6.0 ± 0.3 μm), hyaline, ellipsoidal to
fusiform, often flattened on one side, aseptate, wall 0.5–0.6 μm
thick, smooth or with a thin fibrillar sheath visible in India ink or
with the SEM, filled with one large or numerous small drops.
Asexual morph: unknown.
Culture characteristics: Colonies on MLA 19–24 mm diam after
14 d (22–25 mm after 21 d, 26–29 after 28 d) at 25 °C, circular,
raised. Aerial mycelium abundant, cottony, margins filamentous,
colony surface olive-grey in the centre, dark olivaceous-grey to-
wards margin; reverse black. Colonies on PCA 14–16 mm diam
after 14 d (16–18 mm after 21 d, 18–19 mm after 28 d) at 25 °C,
circular, raised. Aerial mycelium abundant, cottony, margins fila-
mentous, colony olive-grey with dark olivaceous-grey to almost
black margin; reverse black. Sporulation not observed.
Specimens examined:Czech Republic, Southern Bohemia,
Sumava Mts. Na-
tional park,
Zelezn
a Ruda, glacial cirque of the
Cern
e jezero lake, on decaying
wood of Fagus sylvatica, 13 Aug. 1999, M. R
eblov
a M.R. 1586. New Zealand,
South Island, West Coast, Buller Distr., Victoria Forest park, Palmers Hut 18 km
SW of Springs Junction on unpaved road, Lake Christabel track, on decaying
wood of Nothofagus sp., 28 Feb. 2003, M. R
eblov
a M.R. 2671/NZ 160; ibid.,
Westland Distr., Saltwater Forest, Poerua River valley 28 km W of Hari Hari, on
dried driftwood, 12 Mar. 2003, M. R
eblov
a M.R. 2796/NZ 306 (PDD 110879,
culture ICMP 15147).
Notes: The holotype was not accessible for study (pers. comm.,
Wenfeng Zhang, IFRD). The ascospores of T. fibrosa were
described to have a thin fibrillar sheath visible in India ink or with
the SEM (Hyde et al. 2000). The ascal apex and ascospores of
T. fibrosa from a non-holotype specimen were studied by Lee
et al. (2004) at the ultrastructural level. Some ascospores
were surrounded by an electron-transparent mucilage-like matrix
clearly visible towards the end of ascospores when they were still
inside the ascus. Moreover, they detected an additional wall layer
inside the mesosporium, which is also present in the ascospores
of T. biatriispora, but absent in the morphologically similar genus,
Annulatascus.
Our specimens from New Zealand and the Czech Republic fit
well the description and illustration of T. fibrosa from Hong Kong,
China, provided by Hyde et al. (2000), except that the ascospores
do not possess a fibrillar sheath. The occurrence on water-
saturated decayed wood (ICMP 15147, collected as dried drift-
wood on the shores of a river) is consistent with the ecology of
other freshwater species classified in this genus. The material
from the Czech Republic was collected in the glacial cirque in the
Sumava Mts., a unique locality with specific microclimatic condi-
tions. The ascomata in our collections were larger than ascomata
in the type collection (135–255 μmdiamfide Hyde et al. 2000).
The material collected in New Zealand was isolated in axenic
culture, but only sterile dematiaceous mycelium was produced.
Considering that cryptic speciation appears to be common in
this lineage, the morphological differences in the ascospore wall,
ecological differences, and the absence of molecular data from
collections originating from the area of original description of
T. fibrosa, the present collections are tentatively identified as this
species, pending further investigations of accessions from the
area of its description.
Torrentispora fusiformis Fryar & K.D. Hyde, Cryptog. Mycol.
25: 256. 2004.
Notes: For description and illustration see Fryar & Hyde (2004).
This species is known only from Brunei on submerged wood in
brackish and freshwater environments. Torrentispora fusiformis
closely resembles T. calembola in the morphology of the asco-
spores and immersed ascomata with upright cylindrical necks,
but differs in having longer ascospores exceeding 30 μm, and
narrower asci.
Torrentispora novae-zelandiae R
eblov
a & A.N. Mill., sp. nov.
MycoBank MB821778. Fig. 25.
Fig. 23. Torrentispora calembola and T. dubia.A–I. Torrentispora calembola.A–C. Ascomata (with ascoma primordium on A). D, E. Asci. F. Longitudinal section of the ascomal
wall. G. Apical annulus. H. Paraphyses. I. Ascospores. J–S. Torrentispora dubia.J–L. Ascomata. M, N. Asci. O. Longitudinal section of the ascomal wall. P. Apical annulus. Q.
Ascogenous hyphae. R, S. Ascospores. A–C from PRA-12745, D –I from PRA-12744, J –R from PRA-12746, S from PAD. Scale bars: A–C, J–L = 250 μm, D, E, H, M, N,
Q=10μm, F, O = 20 μm, G, I, P, R, S = 5 μm.
SORDARIOMYCETES
www.studiesinmycology.org 45
Etymology: Referring to New Zealand, the country where the
fungus was collected.
Ascomata immersed, with only tips of necks protruding, solitary.
Venter 350–450 μm diam, 350 –420 μm high, subglobose, up-
right or lying horizontally on the host, dark brown to black,
glabrous, with brown, septate hairs ca. 2.5 μm wide, sparsely
growing at the base. Neck central, 100–110 μm wide, up to
600 μm long, cylindrical, upright or slightly decumbent, apex
without sulcations. Ostiole periphysate. Ascomatal wall fragile,
25–33 μm thick, two-layered; outer layer consisting of thick-
walled, brown, polyhedral cells of textura prismatica with opa-
que walls; cells tending to be darker towards the outside,
becoming flattened and paler towards the interior. Inner layer
consisting of several rows of thin-walled, hyaline, flattened cells.
Paraphyses abundant, longer than the asci, becoming dis-
integrated with age, septate, slightly constricted at the septa,
hyaline, 4.5–6.5 μm wide, tapering to 2–2.5 μm. Asci (170–)
200–213(–225) × 8.5–10.5 μm (mean ± SD = 203 ± 18.3 ×
9.5 ± 0.9 μm), 140–167(–197) μm (mean ± SD = 166.3 ±
18.6 μm) long in the sporiferous part, truncate at the apex, cy-
lindrical, stipitate with 8 uniseriate ascospores, apical annulus
Fig. 24. Torrentispora fibrosa.A–D. Ascomata. E. Longitudinal section of the ascomal wall. F–I. Asci (H, I in India ink). J. Apical annulus. K, L. Ascospores (L in India ink). M.
Paraphyses. N, O. Colonies on MLA and PCA after 28 d. A–M from PDD 110879, N, O from ICMP 15147). Scale bars: A–D = 250 μm, E = 20 μm, F –I, M = 10 μm, J, K, L = 5 μm.
R
EBLOV
AET AL.
46
Fig. 25. Torrentispora novae-zelandiae.A–C. Ascomata. D. Longitudinal section of the ascomal wall. E, F. Asci. G. Apical annulus. H–J. Ascospores. K. Ascogenous hyphae.
L. Paraphyses. M, N. Colonies on MLA and PCA after 28 d. A–L from PDD 110880, M, N from ICMP 18368. Scale bars: A–C = 250 μm, D = 20 μm, E, F, J–L=10μm,
G–I=5μm.
SORDARIOMYCETES
www.studiesinmycology.org 47
5–5.5 μm wide, 4–4.5 μm high. Ascospores (17.5–)18–25(–26)
×(6–)7–8.5 μm (mean ± SD = 22.9 ± 3.2 × 7.4 ± 0.7 μm), hy-
aline, fusiform, often flattened on one side, aseptate, smooth-
and thick-walled (0.5–0.6 μm), filled with a large drop or
numerous small drops.
Culture characteristics: Colonies on MLA 20–23 mm diam after
14 d (24–26 mm after 21 d, 27–30 after 28 d) at 25 °C, circular,
raised. Aerial mycelium abundant, present mostly in the centre of
the colony, cottony, margins filamentous, colony surface pale
grey to whitish in the centre, dark olivaceous-grey towards
margin formed by substrate mycelium with a black hue; reverse
black. Colonies on PCA 11–14 mm diam after 14 d (12–15 mm
after 21 d, 15–18 mm after 28 d) at 25 °C, circular, raised. Aerial
mycelium abundant, cottony, margins filamentous, colony sur-
face beige-brown in the centre, dark olivaceous-grey towards
margin; reverse black. Sporulation not observed.
Specimens examined:New Zealand, South Island, West Coast, Buller Distr.,
Victoria Forest Park, Big River Inanganua track, 14 km SE of Reefton, on
decaying wood, 5 Apr. 2005, M. R
eblov
a M.R. 2956/NZ 719 (holotype, PDD
110880, culture ex-type ICMP 18368); ibid., M.R. 3438/NZ 718.
Notes:Torrentispora novae-zelandiae is similar to T. calembola in
having long-necked immersed ascomata and asci of similar
length, but differs from the latter in narrower asci and shorter
ascospores. The ascospores in the non-type collection (M.R.
3438/NZ 718) are 19–22 × (5.5–)6–7μm, the asci are
199–210 × 8.5–9.5 μm.
Torrentispora pilosa Shearer & F.R. Barbosa, Mycologia 105:
339. 2013.
Notes: For description and illustration see Barbosa et al. (2013).
Torrentispora pilosa is known from submerged wood in a tropical
stream in Costa Rica and is similar to T. fusiformis in ascospore
morphology, but differs from the latter in having larger, hairy
ascomata, shorter asci and ascospores that can develop up to
two septa.
DISCUSSION
Soon after Ceratostomella was established with the simple
diagnosis “Perithecia et asci Ceratostomatis. Sporidia continua,
hyalina”(Saccardo 1878a), it became a large, heterogeneous
group of morphologically similar species whose highly poly-
phyletic nature was revealed with molecular data (e.g. R
eblov
a
2006, 2011, Huhndorf & Fern
andez 2005, Huhndorf et al.
2008, 2009, R
eblov
a&
St
ep
anek 2009, de Beer et al. 2013a,
b, 2014, R
eblov
aet al. 2015a, b). These studies have chal-
lenged the traditional divisions separating species in this fungal
complex based on characters of ascoma anatomy, ascospore
morphology (colour), branching pattern of ascogenous hyphae,
and conidiogenesis. The “Ceratostomella phenotype”apparently
represents an ecological adaptation, and so it has evolved
numerous times throughout the Sordariomycetes.
Our efforts to clarify phylogenetic relationships among taxa in
this fungal complex focused on those Ceratostomella species
with hyaline or pale brown ascospores and persistent asci that
show similarity to Lentomitella and Torrentispora. Based on
revision of their holotypes, recently collected material, living
cultures and novel DNA sequences, Ceratostomella dubia,
C. fuscolutea,C. hyalostoma and C. subdenudata were
confirmed to be distantly related to Ceratostomella and were
transferred to three other genera in this study. Furthermore,
revision of holotypes of other Ceratostomella species resembling
Lentomitella, i.e. C. investita,C. maderensis,C. triseptata and
C. vestita var. varvicensis, confirmed their placement in Lento-
mitella. Other species such as C. cirrhosa and C. crinigera had
already been allocated to Lentomitella (R
eblov
a 2006). The
taxonomic status of other Ceratostomella spp. with persistent
asci, their published details, and where available, phylogenetic
data, are listed below.
The combined six-gene phylogenetic analysis of these
Ceratostomella spp., the ex-type strains of Fusoidispora aquatica
and Pseudodiplococcium ibericum, non-type specimens of
Pseudoannulatascus biatriisporus,Spadicoides bina and Xen-
ospadicoides atra, and several other undescribed taxa revealed
a robust, strongly-supported monophyletic clade (Fig. 1). It
represents the Xenospadicoidales and contains four genera, i.e.
Calyptosphaeria,Lentomitella,Spadicoides and Torrentispora.It
is embedded in a large subclade (70/0.99) within the Sordar-
iomycetidae comprising Atractosporales,Papulosaceae,Spor-
idesmiaceae and numerous genera (as incertae sedis) with a
prevailing mode of holoblastic conidiogenesis (Fig. 2).
Hern
andez-Restrepo et al. (2017) introduced Xen-
ospadicoidales based on partial nucLSU sequence data for two
monotypic dematiaceous hyphomycete genera Xenospadicoides
and Pseudodiplococcium. A part of this order was introduced by
Zhang et al. (2017) as the monotypic family Lentomitellaceae for
L. cirrhosa and L. crinigera based on existing ITS, nucLSU,
nucSSU and rpb2 sequence data (R
eblov
a 2006, R
eblov
aet al.
2016). However, the sampling in both studies was insufficient,
either members of Lentomitella or Spadicoides were absent from
the phylogenetic analyses and the relationship between Lento-
mitella and Torrentispora (as P. biatriisporus and F. aquatica)in
Zhang et al. (2017) was not statistically supported. Based on
results of our ML and BI analyses of six- and three-gene datasets
and in accordance with the principle of priority, the monotypic
Lentomitellaceae (Zhang et al. 2017) is synonymised with the
Xenospadicoidaceae (Hern
andez-Restrepo et al. 2017).
The Xenospadicoidales phylogenetic tree (Fig. 1) revealed a
topology that is consistent with ascospore morphology and
conidiogenesis shared by members of this order. These taxa are
characterised by non-stromatic, dark ascomata with a central,
cylindrical or rostrate neck with or without sulcations at the tip,
persistent asci with a distinct, non-amyloid apical annulus and
partially disintegrating paraphyses. The asexual morphs are
dematiaceous hyphomycetes with sympodially proliferating
holoblastic conidiogenous cells, phaeoisaria-like in Lentomitella,
or with a tretic mode of asexual spore production in Spadicoides
accompanied by holoblastic-denticulate conidiogenesis of the
selenosporella-like synasexual morph. Other members of the
order produce mostly sterile mycelium in axenic culture, but their
ascospores often do not germinate in vitro. They are cosmo-
politan and occur on decaying wood, bark and other plant ma-
terial in freshwater and terrestrial habitats.
The morphology of ascospores, and to a certain extent also of
the ascomatal neck, are the main diagnostic features to distin-
guish genera in the Xenospadicoidales.InCalyptosphaeria the
ascospores are dull brown prior to discharge, smooth-walled,
ellipsoidal to fusiform with a tendency to collapse within the
R
EBLOV
AET AL.
48
asci, while the ascomatal neck is sulcate or rarely roughened
without sulcations. In Lentomitella the ascospores are hyaline,
longitudinally striate, ellipsoidal or ellipsoidal-fusiform, and the
neck is sulcate. In Torrentispora the ascospores are hyaline,
smooth-walled or occasionally with a fibrillar sheath, distinctly
thick-walled, fusiform or elongated fusiform, rarely cymbiform,
and the neck is smooth or roughened without sulcations. The
sexual morphs of Spadicoides have hyaline, ellipsoidal to ovoid
ascospores; in species with verruculose ascospore walls the
ascomatal neck is partly light fulvous to subhyaline without sul-
cations (S. hyalostoma), or the ascospores are smooth-walled
and the ascomatal neck is dark and sulcate (S. bina,
S. fuscolutea). In all genera except Lentomitella we observed a
delayed formation of septa in aseptate ascospores within or
outside the asci. In Lentomitella the ascospores are distinctly
1–3-septate early in ontogeny. In addition to the ascospore and
ascomatal neck morphology, the apical annulus also appears to
have taxonomic value. In all genera it is distinct, refractive, but
differs in size. The relatively smallest apical ring occurs in Len-
tomitella (2.5‒3 × 1.5‒2μm, width × height) and Spadicoides
(2‒3.5 × 1.5‒2μm), medium-sized apical rings occur in
Calyptosphaeria (3‒5 × 2.5‒4.5 μm) and the largest apical rings
are present in members of Torrentispora (4‒8 × 3.5‒4.5 μm).
Calyptosphaeria
Calyptosphaeria,typified with C. tenebrosa, is segregated from
Lentomitella based on evidence from phylogenetic analyses and
morphology of ascospores. The genus comprises three other
species, i.e. C. collapsa,C. subdenudata (Peck 1879, Barr 1986,
as L. pallibrunnea in Huhndorf et al. 2008) and C. tropica (as
L. tropica in Huhndorf et al. 2008). The asexual morph is unknown
and since the ascospores do not germinate in vitro, all sequence
data were generated from DNA extracted directly from herbarium
material (Huhndorf et al. 2008 and this study). Members of
Calyptosphaeria inhabit decaying wood and bark and have been
reported from terrestrial or rarely freshwater biotopes in tropical
and temperate zones of southern and northern hemispheres.
Calyptosphaeria is closely related to Lentomitella, which dif-
fers by hyaline, longitudinally striate ascospores. Calyp-
tosphaeria also shows a certain resemblance to Xylomelasma
(R
eblov
a 2006) in having ascomata with a sulcate neck and
brown smooth-walled ascospores, but the latter genus differs by
aseptate, slightly apiculate ascospores and presence of discrete
ascogenous cells, which simultaneously produce several lateral
and terminal dehiscent cells from which asci arise. Based on
DNA sequence data, Calyptosphaeria and Xylomelasma are
unrelated (Fig. 2).
Lentomitella
New collection data, living cultures and phylogeny based on
novel DNA sequences of six nuclear ribosomal and protein-
coding loci revealed a strongly supported Lentomitella clade
(Fig. 1). Its members occur on decaying wood and bark in
terrestrial habitats in temperate zones of both hemispheres. They
are widely distributed on hardwoods, occasionally on wood of
fruit trees (L. conoidea,Feltgen 1903) or senescent flower heads
of Protea lepidocarpodendron (L. unipretoriae,Marincowitz et al.
2008), and some species like L. crinigera exhibit a clear pref-
erence for coniferous wood.
Routine sequencing of ITS and in-depth comparative analysis
of the ITS2 2D structure revealed a novel genetic variation
among Lentomitella isolates. Eleven species are accepted in the
genus, nine of which are included in our phylogenies. No DNA
sequence data or cultures of L. investita (Schweinitz 1832) and
L. unipretoriae (Marincowitz et al. 2008) exist. Based on results
from phylogenetic analyses and revision of morphological
characters, three species are excluded from Lentomitella.Len-
tomitella pallibrunnea and L. tropica (Huhndorf et al. 2008) with
pale brown, aseptate, smooth-walled ascospores are transferred
to Calyptosphaeria as C. subdenudata and C. tropica, while
L. tomentosa (R
eblov
a 2006) with hyaline, aseptate and smooth-
walled ascospores is transferred to Spadicoides as S. fuscolutea.
The asexual morphs of Lentomitella spp. are phaeoisaria-like
dematiaceous hyphomycetes formed only in vitro. Interestingly,
sporulation was observed only in collections of L. investita,
L. sulcata and Lentomitella sp., all originating from New Zealand.
Lentomitella sulcata and Lentomitella sp. belong to the same
subclade and produce globose to ellipsoidal conidia on minute
but conspicuous denticles. The asexual morph of L. investita
produces clavate to obovate conidia on a long rachis containing
numerous but indistinct denticles. The cultures derived from
ascospores of European collections remain sterile.
The main diagnostic feature to distinguish among species of
Lentomitella is the morphology of ascospores. The ascospores
are 1–3-septate, often with a delayed formation of the second and
third septum prior to discharge, which makes the correct identi-
fication sometimes difficult. Therefore, it is important to examine
as many ascospores as possible and also look for old shrinking
ascospores released from the asci, which may contain additional
septa. Only L. vestita, the type species, has truly 1-septate as-
cospores. Other species such as L.crinigera,L. magna,
L. striatella,L. sulcata and L. unipretoriae have ascospores 3-
septate early in ontogeny. Alternatively, L. conoidea,L. investita
and L. tenuirostris often develop 1(–2)-septate ascospores with
the third septum delayed and sometimes not formed at all.
The presence of longitudinal ridges in ascospores is a good
character to distinguish this genus from other morphologically
similar taxa. The ridges are usually shallow, but well visible,
however two interesting cases were observed. In ascospores of
L. investita and L. vestita the individual ridges are more con-
spicuous than in other species and often can be seen protruding at
the poles. In L. magna the longitudinal ridges sometimes become
discontinuous giving the ascospore wall a reticulate appearance.
Ascomatal morphology is highly variable within collections of
the same species and among collections of different species, and
thus, is of limited value for distinguishing species. Ascomatal
characters include the degree of immersion in the wood or bark,
presence and abundance of hairs growing from the venter and
elongation of the neck and its position (upright or slightly
decumbent). These characters are often influenced by humid
conditions; for example, longer necks often develop when
ascomata grow in cracks in wood or are positioned under the
bark. The neck can sometimes appear slightly wider at the tip,
which is often caused by a rupture of the deeply sulcate ostiolum.
In some cases, the sulcations may disappear upon aging,
leaving the surface of the neck roughened.
The longitudinally striate, hyaline ascospores of Lentomitella
resemble those of Phomatospora (Barr 1994, Cai et al. 2006),
but the latter taxon differs by ascomata developed under a thin
clypeus, aseptate ascospores and occurrence on damp or
submerged wood or herbaceous material.
SORDARIOMYCETES
www.studiesinmycology.org 49
Lentomitella and the CBC species concept
The study of interspecific relationships of Lentomitella is
corroborated by morphology, phylogenetic data and the ITS2 2D
structure using the CBC species concept (Coleman 2000, 2003,
2007, Coleman & Vacquier 2002). A less conservative modifi-
cation to this concept was proposed by Müller et al. (2007).
According to these authors any CBC in the ITS2 is informative.
Although the multicopy nature of ITS2 sequences may pose a
potential danger of existence of intragenomic CBC and executing
the CBC species concept, Wolf et al. (2013) demonstrated that
the probability that there is no intragenomic CBC is ~0.99.
Species delimited by CBCs are further characterised by hCBCs
and non-CBCs. However, genotypes of two organisms differing
by a single hCBC indicate that they can theoretically interbreed.
The rapidly evolving hCBCs and short-lived non-CBCs sub-
stitutions occur more frequently than CBCs and may facilitate
faster ecological adaptations of organisms followed by changes
in morphology (Caisov
aet al. 2011, R
eblov
aet al. 2013, 2015a).
The identification of canonical pairs that undergo reciprocal
substitution (C=G 4G=C, A-U, U-A) in helices I–III aided us in
discriminating among species of Lentomitella. The distinction of
Lentomitella species is supported by seven CBCs (Figs 3, 4).
The 14th base pair of the helix III of ITS2 is a particularly
interesting site because changes among Lentomitella spp.
occurring at this position include the full evolution of the recip-
rocal substitutions involving CBC, hCBC and non-CBC such as
U-A /G/A /G/U /G=C. Furthermore, in search for
hCBCs and non-CBCs we identified ten such events on helix II,
all linked to species originating from New Zealand (L. cirrhosa,
L. magna,L. striatella,L. sulcata) except one hCBC unique for
L. crinigera and L. obscura both from European material.
The CBCs identified in helices I and III support the distinction of
all Lentomitella species except the two species pairs, i.e.
L. conoidea and L. tenuirostris,L. cirrhosa and L. striatella.They
illustrate a situation when two closely related species are distin-
guished in the absence of a CBC between them. A close rela-
tionship between L. conoidea and L. tenuirostris is strongly
supported by individual ITS, tub2, and rpb2 sequence data and by
the combined analysisof all six genes. There are only a fewsubtle
characters of ascomata, asci, ascospores and macroscopic colony
characters that distinguish these two species, which make their
correct identification in the absence of molecular data challenging.
Their ITS sequences show 94 % similarity, but the differences
between their sequences are not associated with helices I–III of the
ITS2. A second case was observed between L. cirrhosa and
L. striatella, which also lack a CBC between them. Both species
originate from New Zealand and are positioned at the base of the
Lentomitella clade. Lentomitella striatella differs from L. cirrhosa in
longer and wider asci and slightly longer 3-septate ascospores,
while ascospores of L. cirrhosa are predominantly 1-septate and
additional septa develop later and are rarely visible in old asco-
spores. Their ITS sequences show 96 % similarity.
The third interesting case concerns three morphologically
highly similar species, L. obscura,L. sulcata and Lentomitella sp.
that form a monophyletic clade, which is delimited from other
species by a CBC on the 8th base pair of the helix III of the ITS2
(Fig. 4). Although L. obscura and L. sulcata are further distin-
guished by a unique CBC between them, the distinction between
L. obscura represented by three European strains and Lento-
mitella sp. based on a single New Zealand strain is not supported
by any CBC. Their ITS sequences show 97 % similarity. The only
difference between them is the occurrence of a non-CBC in the
13th base pair of the helix III in the ITS2.
Spadicoides
Spadicoides, typified by S. bina, was introduced by Hughes
(1958) for a group of dematiaceous hyphomycetes occurring
as saprobes on decaying wood or plants remnants. For the first
time we show the sexual-asexual morph relationship between
Spadicoides and perithecial ascomycetes. Hughes (1958)
considered unbranched conidiophores as one of the key diag-
nostic characters to separate Spadicoides from the morpholog-
ically similar Diplococcium. His concept was adopted by Ellis
(1963,1971), Holubov
a-Jechov
a (1982) and Wang (1976).
Sinclair et al. (1985) considered the formation of single vs.
catenate conidia as the main diagnostic criterion, superior to the
simple/branched conidiophores, to distinguish Spadicoides from
Diplococcium. He abandoned the generic concept of Spadi-
coides sensu Hughes (1958) and transferred four Spadicoides
species with conidia formed regularly or occasionally in short
chains to Diplococcium.
Using partial nucLSU sequences, Shenoy et al. (2010) sug-
gested that Spadicoides and Diplococcium are polyphyletic and
unrelated to each other. In the same nucLSU phylogeny (Shenoy
et al. 2010), the strain CBS 113708 of S. bina (nucLSU sequence
EF204507) occurred in the Cordanales as sister to Cordana
pauciseptata (as Porosphaerella cordanophora, the sexual
morph; strain M.R. 1150, nucLSU sequence AF178563) with
100 % bootstrap support, while morphologically similar Spadi-
coides atra CBS 489.77 was shown as sister to Lentomitella.
Spadicoides bina and C. pauciseptata share 1-septate, brown,
ellipsoidal conidia of comparable size born terminally or laterally
on upright, macronematous, dematiaceous conidiophores, but
they differ in the mode of conidiogenesis. It is tretic in Spadi-
coides while in Cordana conidia are borne on minute denticles
from intercalary and terminal swellings. The examination of a
specimen [Sweden, Uppland, Dalby par., Jerusalem, on decay-
ing wood of Picea abies, 17 Apr. 1986, K. & L. Holm 3980, F-
540504 (UPS)], used for isolation of the “S. bina”strain CBS
113708 and preparation of the dried culture [(1992.02) F-540502
(UPS)] revealed that the fungus present on the wood and in the
dried culture is C. pauciseptata (Fig. 26). It is obvious that the
fungus was originally misidentified, which was merely followed by
Shenoy et al. (2010) and Hern
andez-Restrepo et al. (2017), who
segregated S. atra from Spadicoides into Xenospadicoides.It
was distinguished from morphologically similar Pseudodiplo-
coccium by arrangement of conidia, solitary in Xenospadicoides
and catenate in Pseudodiplococcium.
Spadicoides bina was recently recollected and obtained in
axenic culture (CBS 137794) from isolated ascospores of an
undescribed lentomitella-like species (PRA-13420). A strongly
supported Spadicoides clade containing S. bina,S. fuscolutea,
S. hyalostoma and X. atra was recovered in our six-gene phy-
logeny (Fig. 1) and ITS-nucLSU analysis (Supplementary Fig. 1),
which included also P. ibericum. Based on molecular DNA data
and morphology of conidia, conidiogenous cells and co-
nidiophores, X. atra and P. ibericum are accepted in Spadicoides;
a new combination is proposed for the later species, and
Pseudodiplococcium and Xenospadicoides are synonymised
with Spadicoides. Based on these results, the generic description
R
EBLOV
AET AL.
50
of Spadicoides is emended to include both sexual and asexual
morphs. The genus is characterised by ascomata with long
necks, hyaline, aseptate or 1-septate, delicately verruculose or
smooth-walled ascospores, unbranched or branched co-
nidiophores and dark brown conidia formed singly or in a chain,
and the selenosporella-like synasexual morph.
Selenosporella is a hyphomycete genus (Sordariomycetes,
incertae sedis) with pale brown to subhyaline conidiophores
producing usually clavate, fusiform, obclavate to falcate, hyaline
conidia (MacGarvie 1968). However, the exact mode of con-
idiogenesis has been the subject of a broad discussion. Although
MacGarvie (1968) described the conidiogenesis of S. curvispora,
the type species, as holoblastic-denticulate with conidia arising
sympodially in basipetal succession from minute denticles, Ellis
(1971) and Hughes (1979) interpreted the conidiogenesis as
phialidic or possibly phialidic based on study of other
Selenosporella and selenosporella-like species. Onofri &
Castagnola (1982) studied S. curvispora with electron micro-
scopy and concluded that the conidiogenesis is holoblastic-
denticulate. It is likely that selenosporella-like asexual morphs
or synasexual morphs occuring in various taxonomic groups
have different modes of conidiogenesis, which is difficult to
observe with light microscopy.
The selenosporella-like synasexual morph observed in 4–8-
wk-old axenic cultures of S. bina and S. fuscolutea also was
reported for other species of Spadicoides, i.e. S. heterocolorata
(Casta~
neda et al. 1997), S. obclavata (Kuthubutheen & Nawawi
1991a) and S. wufengensis (Li et al. 2010). Furthermore, it was
described as a synasexual morph of dematiaceous hyphomy-
cetes such as Ceratosporium (Hughes 1964), Teratosperma
(Hughes 1951, Matsushima 1975), and also Diplococcium, e.g.
D. hughesii (Wang & Sutton 1998), D. dimorphosporum and
Fig. 26. Cordana pauciseptata.A, B. Conidiophores. C–E. Conidia. A –C from dried culture F-540502 (UPS), D, E from F-540504 (UPS). Scale bars: A –E=10μm.
SORDARIOMYCETES
www.studiesinmycology.org 51
D. singulare (Hern
andez-Restrepo et al. 2012). The
selenosporella-like synasexual morph was reported also for
Endophragmiella, e.g. E. dimorphospora (Awao & Udagawa
1974, Matsushima 1975), E. subolivacea (Matsushima 1975),
and E. theobromae (Hughes 1979), including Endophragmiella
spp. linked with sexual morphs such as Echinosphaeria can-
escens and Lasiosphaeria punctata (Hughes 1979, Sivanesan
1983)orRuzenia spermoides (Gams 1973, Miller & Huhndorf
2004b). Oxydothis selenosporellae (Samuels & Rossman
1987) and Iodosphaeria (Samuels et al. 1987) are additional
sexual morphs linked with a selenosporella-like asexual morph.
Fungi with a selenosporella-like phenotype were described as
part of the life cycle of several other dematiaceous hyphomy-
cetes such as Acrodictys bambusicola (Matsushima 1975),
Arachnophora excentrica (Hughes 1979), Polytretophora cal-
carata (= Spadicoides calcarata,Kuthubutheen & Nawawi
1991b) and Quadracaea mediterranea (Lunghini et al. 1996).
The position of Spadicoides verrucosa (Rao & de Hoog 1986)
is in agreement with Shenoy et al. (2010). In our analysis this
species is also placed in the Sordariomycetidae but in a separate
clade near members of the Phomatosporales and Magnapor-
thales. According to Shenoy et al. (2010) Spadicoides xylogena
is related to Curvularia brachyspora (ATCC 58872) (Pleospor-
ales,Dothideomycetes); these fungi show remarkable similarity.
Considering the obvious polyphyly of Spadicoides, the genus
requires taxonomic revision.
The asexual morph of Spadicoides is most similar to Dip-
lococcium in pigmented, macronematous conidiophores, polyt-
retic conidiogenous cells and dark conidia, but it differs in having
mostly unbranched or rarely branched conidiophores and is
linked to morphologically different sexual morphs. Diplococcium
has been linked to five Helminthosphaeria species (Helmin-
thosphaeriaceae,Sordariomycetes) as a presumed asexual
morph based on juxtaposition of conidiophores and ascomata
(Samuels et al. 1997, R
eblov
a 1999), while Diplococcium spi-
catum, the type species, was positioned in the Helotiales
(Leotiomycetes) based on molecular DNA data (Shenoy et al.
2010).
Torrentispora
Torrentispora, typified by T. fibrosa, was erected as a monotypic
genus in the Annulatascaceae based on its massive, non-
amyloid apical annulus (Hyde et al. 2000). It was introduced
for taxa morphologically similar to Annulatascus, but distinct in
ascomatal wall having irregular rows of cylindrical cells in surface
view and smaller ascospores (< 20 μm) vs. textura epidermoidea
in surface view and larger ascospores (> 20 μm) in Annulatascus
(Hyde 1992). Torrentispora was further distinguished from
Annulatascus by morphology of the ascospores at the ultra-
structural level; the ascospores of Torrentispora lack episporial
verrucose ornamentation and possess an additional wall layer
inside the mesosporium (Lee et al. 2004).
In the phylogeny based on three nuclear markers (Fig. 2),
Torrentispora is shown unrelated to Annulatascus, but forms a
monophyletic, strongly supported subclade nested in the Xen-
ospadicoidales. To date, the genus comprises four species
mostly from freshwater habitats from subtropical, tropical and
temperate zones, i.e. T. crassiparietis,T. fibrosa,T. fusiformis
and T. pilosa (Hyde et al. 2000, Fryar & Hyde 2004, Barbosa
et al. 2013). Based on the evidence from DNA sequence data
we introduce T. calembola and T. novae-zelandiae as new
species mainly from terrestrial habitats and we propose three
new combinations (T. aquatica,T. biatriispora and T. dubia).
Torrentispora fibrosa was originally collected on submerged
wood in streams in Hong Kong, China where the climate is
subtropical tending towards temperate for nearly half of the year
(Hyde et al. 2000, Ho et al. 2001). Other collections have been
reported from Florida USA (Raja et al. 2003), but no states north
of Florida (Shearer 1993, 2001). Our collections originate from
temperate zones of Europe and New Zealand, but differ from the
type species by absence of the fibrillar sheath in ascospores.
Ingold (1966) speculated on the latitudinal distribution of aquatic
hyphomycetes by examining aquatic spores in samples of
stream foam at a distance of every 15° latitude. Conversely, it
has been supported by molecular data that some freshwater
ascomycetes are not restricted to a certain latitude, e.g. collec-
tions of the freshwater species Annulusmagnus triseptatus
(Annulatascales) from Canada, France, Hong Kong and
Venezuela form a strongly supported monophyletic clade
(Campbell & Shearer 2004, Dayarathne et al. 2016).
Pseudoannulatascus was introduced as a monotypic genus
(Luo et al. 2015) to include Annulatascus biatriisporus (Hyde
1995) based on partial nucLSU sequence data. Pseudoannula-
tascus biatriisporus is most similar to Torrentispora in the
morphology of ascomata, asci and ascospores. Their close
relationship was revealed in our phylogeny using DNA se-
quences from a non-type collection of P. biatriisporus from Costa
Rica [specimen A 464-3, Raja et al. (2003)], our collection of
T. fibrosa from New Zealand (ICMP 15147) and other Torren-
tispora species. Therefore, Pseudoannulatascus is synonymised
with Torrentispora.
The monotypic genus Fusoidispora was described for a
freshwater lignicolous species, F. aquatica (Vijaykrishna et al.
2005). We analysed the partial nucLSU sequence of
F. aquatica [AY780365, holotype HKU(M) 17484, Vijaykrishna
et al. (2005)] in the nucLSU (not shown) and six-gene (Fig. 1)
phylogenies. This species was nested in the Torrentispora clade
with high statistical support. Despite the obvious difference in the
ascospore morphology, the DNA sequence data of the holotype
suggest that F. aquatica is a member of Torrentispora and
therefore a new combination is proposed.
Torrentispora aquatica is most similar to Pisorisporium
cymbiforme (Pisorisporiales,Sordariomycetidae)(R
eblov
a
et al. 2015c) in morphology and size of ascomata, which lie
horizontally on the host, ascomatal wall, asci and morphology
of long-fusoid to cymbiform, thin-walled ascospores, including
the numerous guttules arranged in a chain within ascospores
like peas in a pod. It differs from P. cymbiforme in ascospores
with globose mucilaginous pads at both ends and the non-
amyloid apical annulus. The DNA of T. aquatica was extrac-
ted directly from herbarium material (Vijaykrishna et al. 2005).
However, we do not exclude the possibility that the DNA was
extracted from another fungus. It is well known that different
species of aquatic fungi occur close to each other on the same
substrate; we often encountered ascomata of Lentomitella,
Spadicoides and Torrentispora next to each other on the nat-
ural substrate.
R
EBLOV
AET AL.
52
REVISION OF CERATOSTOMELLA SPP. WITH
PERSISTENT ASCI
Species of Ceratostomella with evanescent asci that were
transferred to members of Microascales and Ophiostomatales
are listed in de Beer et al. (2013b, 2014). The following list in-
cludes revised Ceratostomella species with persistent asci in
three categories: as accepted species of Ceratostomella s. str.,
species excluded from Ceratostomella and described in other
genera and species of uncertain status. Names in bold refer to
the currently accepted classification and are accompanied by
short notes, reference to full synonymy and additional published
details.
Species accepted in Ceratostomella s. str.
Ceratostomella cuspidata (Fr.) R
eblov
a, Mycologia 98: 77.
2006.
Basionym:Sphaeria cuspidata Fr., Syst. mycol. 2(2): 474. 1823.
Synonym:Ceratostoma cuspidatum (Fr.) Sacc., Syll. fung. 1:
220. 1882.
Ceratostomella pyrenaica R
eblov
a & J. Fourn., in R
eblov
a,
Mycologia 98: 78. 2006.
Ceratostomella rhynchophora (De Not.) R
eblov
a, Mycologia
98: 78. 2006
Basionym:Sordaria rhynchophora De Not., Comm. Soc. crittog.
Ital. 2, Fasc. 3: 480. 1867.
Ceratostomella rostrata (Tode : Fr.) Sacc., Syll. fung. 1: 408.
1882.
Basionym:Sphaeria rostrata Tode, Fung. mecklenb. sel. 2: 14.
1791 : Fr., Syst. Mycol. 2: 473. 1823.
Notes: For full synonymy, descriptions, illustrations, phylogeny,
discussion, nomenclatural notes on C. rostrata and key to
Ceratostomella s. str. see R
eblov
a (2006).
Species excluded from Ceratostomella and
described in other genera
Barbatosphaeria barbirostris (Dufour : Fr.) R
eblov
a, Mycologia
99: 727. 2007.
Basionym:Sphaeria barbirostris Dufour, Turp. Icon. fig. 1. 1820:
Fr., Syst. Mycol. 2: 473. 1823.
Synonyms:Ceratostomella barbirostris (Dufour : Fr.) Sacc., Syll.
Fung. 1: 410. 1882.
Ceratostomella trichina (Pers.) Sacc., Syll. fung. 1: 410. 1882.
Ceratostomella dispersa (P. Karst.) Sacc., Syll. Fung. 1: 411.
1882.
Specimens examined:Germany, Rhineland, Vosges, Wasgau, on decaying
wood of Quercus sp. (syntype of S. trichina Pers. in Litt., Mougeot & Nestler,
Stirp. Crypt. Voges. No. 666, K 147294; C. Roumegu
ere Fung. Sel. Gal. Exs. No.
187, K 147310).
Notes: Revision of the holotypes of Ceratostomella barbirostris
and C. dispersa revealed that these species are conspecific and
were transferred to the new genus Barbatosphaeria as
B. barbirostris (R
eblov
a 2007). For full synonymy, description,
illustration, holotype and other material examined see R
eblov
a
(2007), for phylogeny, additional illustrations and key to Barba-
tosphaeria spp. see R
eblov
aet al. (2015b).
Recently, two collections labelled Sphaeria trichina, with the
same locality and host information, were located in the Kew
fungarium. The specimen labelled as syntype, K 147294,
matches the description of Barbatosphaeria barbirostris. The
other specimen, K 147310, although from the same host, con-
tains a different piece of wood containing empty ascomata.
Calyptosphaeria subdenudata (Peck) R
eblov
a & A.N. Mill.
Basionym:Sphaeria subdenudata Peck, Ann. Rep. N.Y. St. Mus.
nat. Hist. 32: 52. 1880 (1879).
Synonym:Ceratostomella subdenudata (Peck) M.E. Barr, Bull.
N.Y. St. Mus. 459: 44. 1986.
Notes: This study; for full synonymy see above.
Ceratosphaeria lampadophora (Berk. & Broome) Niessl, Verh.
nat. Ver. Brünn 14: 203. 1876.
Basionym:Sphaeria lampadophora Berk. & Broome, Ann. Mag.
nat. Hist., Ser. 3. 3: 372. 1859.
Synonym:Ceratostomella lampadophora (Berk. & Broome)
Cooke, Grevillea 17: 49. 1889.
Notes: For description, illustration and phylogeny of
C. lampadophora see Huhndorf et al. (2008), for notes on its
harpophora-like asexual morph and additional phylogenetic data
see R
eblov
a (2006) and R
eblov
aet al. (2011).Ceratosphaeria,
based on C. lampadophora, is a member of the Magnaporthales
(Sordariomycetes).
Chaetosphaeria longispora (Sacc.) P.M. Kirk, Index Fung. 120:
1. 2014.
Basionym:Ophioceras longisporum Sacc., Syll. fung. 2: 360.
1883.
Synonyms:Sphaeria longispora Ellis, Bull. Torrey bot. Club 6:
135. 1877 non Currey 1859 nec Karsten 1873. (Nom. illegit., Art.
53.1)
Ceratostomella longispora (Sacc.) Cooke, Grevillea 17: 50.
1889.
Lasiosphaeria ellisii M.E. Barr, Mycotaxon 46: 48. 1993.
Chaetosphaeria ellisii (M.E. Barr) Huhndorf & F.A. Fern
andez,
Fung. Diver. 19: 27. 2005.
Notes: For synonymy see Barr (1993), for description, illustration
and phylogeny see Huhndorf & Fern
andez (2005).Sphaeria
longispora (Ellis 1877) is a later homonym of S. longispora Curr.
1859 and S. longispora Karst. 1873. Two replacement names
were published for S. longispora. Barr (1993) introduced
Lasiosphaeria ellisii for this fungus, later transferred to Chae-
tosphaeria by Huhndorf & Fern
andez (2005).Kirk (2014)
considered the first combination of S. longispora in Ophioceras
by Saccardo (1883) as the earliest legitimate name of the taxon
in the same rank (Art. 41.3) in order to replace Sphaeria long-
ispora Ellis. Ophioceras longisporum Sacc. therefore becomes a
basionym for all future combinations. Kirk (2014) proposed a new
combination of O. longisporum in the genus Chaetosphaeria but
erroneously cited S. longispora as the basionym. However, it
does not affect the valid publication of this new combination (Art.
41.8c).
Chaetosphaeria myriocarpa (Fr.) C. Booth, Mycol. Pap. 68: 5.
1957.
Basionym:Sphaeria myriocarpa Fr., Syst. mycol. 2(2): 459.
1823.
Synonym:Ceratostomella stevensonii (Berk. & Broome) Sacc.,
Syll. fung. 1: 412. 1882.
SORDARIOMYCETES
www.studiesinmycology.org 53
Notes: For full synonymy, description, illustration and revision of
the holotype see Booth (1957).
Clohiesia corticola K.D. Hyde, Nova Hedw. 61: 126. 1995.
Synonym:Ceratostomella hyalocoronata Inderb., Mycoscience
41: 167. 2000.
Specimen examined:China, Guangdong Province, Wu Gui Shan, 15 km S of
Zhongshan, on decaying branch submerged in a small stream, 8 Nov. 1998, E. M.
Lea~
no & P. Inderbitzin (holotype, UBC F13874).
Notes: The examination of the holotype of C. hyalocoronata
(Inderbitzin 2000) revealed a fungus that fits well in the
description of Clohiesia (Hyde 1995) and is conspecific with the
generic type C.corticola. Using nucLSU partial sequence data,
Clohiesia, originally treated as a member of the Annulatasca-
ceae, is related to the Sordariales (Raja et al. 2003) and currently
placed there (Kirk et al. 2008).
Daruvedia bacillata (Cooke) Dennis, Belarra 2: 25. 1988.
Basionym:Sphaeria bacillata Cooke, Handb. Brit. Fungi 2: 879.
1871.
Synonym:Ceratostomella bacillata (Cooke) Cooke, Grevillea 17:
50. 1889.
Notes:Dennis (1988) proposed a new monotypic genus, Dar-
uvedia, for Sphaeria bacillata (Cooke 1871). Although he did not
find any ascomata in the holotype, Cooke's habit sketches and
drawings of the ascomata, ascus and ascospores, the only
surviving original elements, enabled him to identify his fresh
material as S. bacillata.Hu et al. (2010) designated Dennis's
material as epitype and provided full synonymy and detailed
description of the species based on additional material. In the
absence of DNA sequence data, Daruvedia is either placed in
the Pyrenulaceae (Pyrenulales,Eurotiomycetes) (as Pleuro-
tremataceae fide Barr 1994, Lumbsch & Huhndorf 2010)oris
classified in Dothideomycetes incertae sedis (Eriksson 2006,
Kirk et al. 2008).
Jattaea echinella (Ellis & Everh.) R
eblov
a, Fung. Diver. 49: 182.
2011.
Basionym:Ceratostomella echinella Ellis & Everh., N. Amer.
Pyren. p. 195. 1892.
Notes: For description, illustration and revision of the holotype
see R
eblov
a (2011).Jattaea is a member of the Calosphaeriales,
well-distinguished from Ceratostomella by hyaline, allantoid as-
cospores in clavate asci lacking an apical annulus and ascomata
usually arranged in small valsoid formations.
Jattaea tumidula (Sacc.) R
eblov
a, Fung. Diver. 49: 186. 2011.
Basionym:Calosphaeria tumidula Sacc., Atti Soc. Veneto-Trent.
Sci. Nat. Padova 4: 77–100 (Fungi Ven. novi Ser. 4: 20) 1875.
Synonym:Ceratostomella mali Ellis & Everh., Proc. Acad. Nat.
Sci. Philad. 42: 225. 1890.
Notes: For full synonymy, description, illustration and holotype
information see R
eblov
a (2011).
Lentomitella cirrhosa (Pers.: Fr.) R
eblov
a, Mycologia 98: 82.
2006.
Basionym:Sphaeria cirrhosa Pers., Syn. Meth. Fung. p. 59. 1801
:Fries, Syst. Mycol. 2: 475. 1823.
Synonym:Ceratostomella cirrhosa (Pers.) Sacc., Syll. fung. 1:
408. 1882.
Notes: See R
eblov
a (2006) and this study; for full synonymy see
above.
Lentomitella crinigera (Cooke) R
eblov
a, Mycologia 98: 83.
2006.
Basionym:Sphaeria crinigera Cooke, Grevillea 1: 156. 1873.
Synonyms:Ceratostomella crinigera (Cooke) Cooke, Grevillea
17: 49. 1889.
Ceratostomella triseptata Petr., Annls mycol. 23: 135. 1925.
Notes: See R
eblov
a (2006) and this study; for full synonymy see
above.
Lentomitella investita (Schw.) R
eblov
a
Basionym:Sphaeria investita Schw., Trans. Amer. Phil. Soc. 2,
Vol. 4: 216. 1834.
Synonyms:Ceratostomella investita (Schw.) Starb€
ack, Bih.
Kongl. Svenska Vet.-Akad. Handl. 19(2): 26. 1894.
Ceratostomella vestita Sacc. var. varvicensis Grove, J. Bot. 23:
131. 1885.
Ceratostomella maderensis Petr., Bot. Jahrb., Beiblt. 142: 98.
1929.
Notes: This study; for full synonymy see above.
Lentomitella vestita (Sacc.) Höhn., Annls mycol. 3: 548. 1906.
Basionym:Ceratostomella vestita Sacc., Michelia 1: 370. 1878.
Notes: This study; for full synonymy see above.
Natantiella ligneola (Berk. & Broome) R
eblov
a, Mycol. Res.
113: 996. 2009.
Basionym:Sphaeria ligneola Berk. & Broome, Ann. Mag. nat.
Hist., Ser. 3. 3: 372. 1859
Synonyms:Ceratostomella ligneola (Berk. & Broome) Cooke,
Grevillea 17: 49. 1889.
Ceratostomella ampullasca (Cooke) Sacc., Syll. Fung. 1: 409.
1882.
Ceratostomella similis Kirschst., Krypt. Flora Brandenb. 7: 245.
1911.
Notes: Revision of the holotypes of Ceratostomella ampullasca,
C. ligneola and C. similis revealed that these species are
conspecific(R
eblov
a&
St
ep
anek 2009). They were transferred
to a monotypic genus Natantiella supported by DNA sequences.
Natantiella ligneola is common on strongly decaying wood and
occurs in temperate zones in the northern and southern hemi-
spheres. The genus is placed in incertae sedis position in the
Sordariomycetes. In phylogenetic analysis, Natantiella forms a
monophyletic, strongly supported clade with members of the
Ophiostomatales (this study), and is unrelated to Ceratostomella
s. str. For full synonymy, description, illustration, phylogeny and
holotype and other material examined see R
eblov
a&
St
ep
anek
(2009).
Phaeoacremonium leptorrhynchum (Durieu & Mont.) D.
Gramaje et al., Fung. Biol. 119: 768. 2015.
Basionym:Sphaeria leptorrhyncha Durieu & Mont., Expl. Sci.
Alg., Fl. Alg
er. 1 (livr. 13). p. 510. 1848.
Synonym:Ceratostomella leptorrhyncha (Durieu & Mont.) Sacc.,
Syll. fung. 1: 412. 1882.
Notes: For full synonymy, illustration and holotype examination
see R
eblov
a (2011).Phaeoacremonium leptorrhynchum, known
only from Algeria on Chamaerops humilis, is one of the few
Phaeoacremonium species that are known only in their sexual
morph and only from herbarium material. Based on the revision
of the holotype, R
eblov
a (2011) suggested that C. leptorrhyncha
and Phaeoacremonium novae-zelandiae (as Togninia) are
R
EBLOV
AET AL.
54
conspecific and proposed a new combination as Togninia lep-
torrhyncha, later transferred to Phaeoacremonium by Gramaje
et al. (2015), the correct name for the holomorph. We prefer to
keep both species separate until representative material of
P. leptorrhynchum is recollected, isolated in axenic culture and
subjected to phylogenetic analysis. Evidence gathered during
our current research calls for caution, when morphologically
highly similar material from different continents is compared.
Phomatospora helvetica H. Wegelin, Mitt. thürgau. naturf. Ges.
12: 173. 1894.
Synonyms:Ceratostomella hydrophila Mouton, Bull. Soc. R. Bot.
Belg. 26: 171. 1887.
Phomatospora moravica Petr., Annls mycol. 22: 55. 1924.
Phomatospora luteotingens J. Fourn. & Lechat, Mycosphere 1:
40. 2010.
Specimen examined:Belgium, Beaufays near Li
ege, on submerged decorticated
wood in a stream (holotype, BR 93866-67).
Notes: Based on characters of immersed to partially erumpent,
globose to conical ascomata 500‒650 μm diam under a thin
black clypeus and with a flattened base, cylindrical asci 121‒
134 × 8‒9μm, 100‒120 μm in the sporiferous part, and uni-
seriate, ellipsoidal, hyaline, aseptate, longitudinally striate as-
cospores (11.5‒)12‒13.5 × 5‒6μm, this species fits well the
description of Phomatospora helvetica and P. moravica (von
Hammer & Scheuer 2008). It was later redescribed as
P. luteotingens by Fournier & Lechat (2010). The yellow stain of
the woody substrate was observed in the holotype of
P. luteotingens and is consistently present in other collections
from France and Spain of this species.
To date, the genus Phomatospora includes 120 epithets ac-
cording to Index Fungorum. It comprises fungi from freshwater,
marine and terrestrial habitats on herbaceous debris but also on
wood. The ascospores are hyaline, usually aseptate with longi-
tudinally striate walls and often with a mucilaginous sheath and/
or bipolar appendages (Barr 1994, Cai et al. 2006). Based on the
habitat and characters of ascospores with and without muci-
laginous appendages or sheathes, the genus might be hetero-
geneous. There has been no monographic study on
Phomatospora and very little is known about its asexual morphs
(Rappaz 1992). Although C. hydrophila is the oldest name for
this species, we refrain from making any formal changes until the
genus Phomatospora can be revised.
Pseudorhynchia polyrrhyncha (Penz. & Sacc.) Höhn., Sber.
Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1. 118: 1206. 1909.
Basionym:Ceratostomella polyrrhyncha Penz. & Sacc., Mal-
pighia 11: 408. 1897.
Notes: This tropical species was originally described from Java
on Elletaria sp. in a monotypic genus. Höhnel (1909) recollected
this taxon in the same locality and on the same host as the type.
The third collection was reported by Samuels & Barr (1997) from
Venezuela on a dead leaf of Heliconia sp. Samuels & Barr (1997)
transferred the genus to the Niessliaceae (Hypocreales). For
description and illustration of the type and other representative
material see Höhnel (1909), Müller & von Arx (1962) and
Samuels & Barr (1997).
Spadicoides fuscolutea (Rehm) R
eblov
a
Basionym:Ceratostomella fuscolutea Rehm, Annls mycol. 6:
320. 1908.
Notes: This study; for full synonymy see above.
Spadicoides hyalostoma (Munk) R
eblov
a
Basionym:Endoxyla hyalostoma Munk, Bot. Tidsskr. 61: 62.
1965.
Synonym:Ceratostomella hyalostoma (Munk) Unter., Mycologia
85: 307. 1993.
Notes: This study.
Togniniella microspora (Ellis & Everh.) R
eblov
a, Fung. Diver.
49: 193. 2011.
Basionym:Ceratostomella microspora Ellis & Everh., Proc.
Acad. nat. Sci. Philad. 45: 444. 1894.
Notes: For description, holotype information, discussion and
comparison with the morphologically similar Flabellascus see
R
eblov
a (2011) and R
eblov
aet al. (2004, 2015a).
Torrentispora dubia (Sacc.) R
eblov
a & A.N. Mill.
Basionym:Ceratostomella dubia (Sacc.) Sacc., Syll. fung. 1:
410. 1882.
Notes: This study; for full synonymy see above.
Wallrothiella congregata (Wallr.) Sacc., Syll. Fung. 1: 455.
1882.
Basionym:Sphaeria congregata Wallr., Fl. crypt. Germ. 4: 786.
1833.
Synonym:Ceratostomella sphaerosperma (Fuckel) Sacc., Syll.
fung. 1: 412. 1882.
Specimen examined:Germany, Johannisberg, on decaying wood of Pinus syl-
vestris, spring (holotype, Fungi Rhen Exs. No. 2013, G).
Notes: For synonymy, description, illustration and phylogeny see
R
eblov
a & Seifert (2004) and Huhndorf et al. (2009). Based on
molecular data, Wallrothiella was placed by Huhndorf et al.
(2009) in the Amplistromataceae (Sordariomycetes).
Ceratostomella species of uncertain status
Ceratostomella albocoronata (Ellis) Sacc., Syll. fung. 2, Add. xxx.
1883.
Basionym:Ceratostoma albocoronata Ellis, Am. Nat. 17: 318.
1883.
Specimen examined:USA, New Jersey, Newfield, Gloucester County, on rotten
wood, Jun. 1882 (holotype, NY).
Notes: The holotype contained several empty ascomata of a
lentomitella-like fungus. They were immersed with short, pro-
truding but mostly broken necks and dark hairs growing from the
venter. According to the protologue (Ellis 1883), asci are cylin-
drical 35 × 7 μm with eight ellipsoidal, hyaline ascospores
7.5–9.5 × 3–3.5 μm with 1–2 drops (as 1–2 nucleate). The
dematiaceous hyphomycete mentioned by Ellis (1883), associ-
ated with ascomata and scattered over the wood, produces
obovate, 2–3-septate, 11.5 × 7.5 μm conidia borne singly on the
apices of upright, brown conidiophores. Only a few conidia and
remnants of conidiophores were present and they belong to
Spadicoides obovata (Cooke & Ellis 1876, Ellis 1963). Based on
the description, C. albocoronata does not match any known
species of Lentomitella. On the other hand, ascospore
morphology and the presence of Spadicoides conidia and co-
nidiophores suggest its relationship with species of the latter
SORDARIOMYCETES
www.studiesinmycology.org 55
genus. For an accurate systematic placement, it would be
necessary to collect representative material to generate DNA
sequences and experimentally prove the connection between the
two morphs. To date, the sexual morph of S. obovata is unknown.
Ceratostomella bambusina Höhn., Sber. Akad. Wiss. Wien,
Math.-naturw. Kl., Abt. 1. 118: 337. 1909.
Specimen examined:Western Malaysia, Indonesia, Java Barat (W. Java), Tji-
bodas, on decaying bamboo culm, 1907–1908 (holotype, FH 00258773).
Notes: The material contained several non-stromatic, partially
erumpent, solitary ascomata with a cylindrical upright neck,
persistent, stipitate asci with a non-amyloid apical annulus and
eight globose, hyaline, aseptate ascospores. Similar ascospores
are a diagnostic feature of Amplistroma and Wallrothiella
(Amplistromatales)(Huhndorf et al. 2009) and also occur in
Woswasia atropurpurea (incertae sedis)(Jaklitsch et al. 2013),
both members of the Sordariomycetidae. While the latter taxon
and Amplistroma produce large stromata, Wallrothiella is non-
stromatic. However, it has a thick, three-layered ascomatal
wall and long-necked ascomata that are solitary or most often
confluent. Ceratostomella bambusina can be compared to
Wallrothiella congregata, the type species, in characters of as-
cospores, asci and partly ascomata, but differs in smaller
ascomata that develop individually on the substrate and much
thinner, two-layered ascomatal walls. DNA sequence data of this
species are necessary to confirm its systematic placement.
Ceratostomella canulata (Preuss) Sacc., Syll. fung. 1: 412. 1882.
Basionym:Sphaeria canulata Preuss, Linnaea 26: 714. 1855.
Specimen:Germany, Hoyerswerda, on decaying wood (holotype).
Notes: The type material could not be located in B (pers. comm.,
B. Hein). The fungus is described with immersed globose
ascomata, protruding cylindrical necks with hyphae growing at
the base, cylindrical asci and hyaline aseptate ascospores.
Based on its description it is not possible to identify the fungus or
convincingly suggest its relationship.
Ceratostomella capillaris (Ellis) Sacc., Syll. fung. 2, Add. xxx.
1883.
Basionym:Ceratostoma capillare Ellis, Bull. Torrey bot. Club 9:
20. 1882.
Specimen examined:USA, New York, Gloucester County, on decaying sterile
catkins of Alnus serrulata, 28 Jun. 1881 (holotype, NY).
Notes: Based on morphology of minute, globose ascomata with a
filiform neck, and asci with a partly evanescent wall containing
eight 2‒3-seriate, hyaline to subhyaline, fusiform, slightly curved,
aseptate ascospores with gelatinous appendages at both end,
this species resembles members of Ophiostoma.
Ceratostomella capilliformis E. Bommer, M. Rousseau & Sacc.,
Syll. fung. 9: 573. 1891.
Specimen examined:Belgium, Groenendael, on decaying wood of Carpinus
betulus (holotype, PAD).
Notes: The type material contained empty ascomata 200 μm
diam with cylindrical, upright or decumbent necks ca. 250‒
300 μm long. According to the protologue (Saccardo 1891), the
asci are 30‒36 × 6‒9μm, clavate, slightly swollen at the base,
paraphyses are present, and ascospores are 8 × 3 μm, ellip-
soidal and later curved. Given the morphology of ascomata, asci
and ascospores this species resembles species of the genera
Barbatosphaeria and Jattaea. The swollen base of the asci may
be interpreted as discrete cells arising from ascogenous hyphae,
which is one of the diagnostic features of Barbatosphaeria
(R
eblov
aet al. 2015b). The bulbose base of the ascus stipe also
occurs in members of Jattaea but it is of a different origin
(R
eblov
aet al. 2015a). Given the size and especially the width of
ascospores, C. capilliformis resembles B. dryina (R
eblov
aet al.
2015b), but the latter taxon differs by larger ascomata
400–600 μm diam, longer asci and 1-septate ascospores. Other
species of Barbatosphaeria have usually 1–2μm wide asco-
spores. Among Jattaea species, C. capilliformis can be
compared to J. discreta (R
eblov
a 2011) in morphology and size
of ascomata with a venter 230–400 μm diam, asci (30–)
35–45(–50) × 6.5–8μm and aseptate, suballantoid ascospores
(6–)6.5–8 × 1.5–2μm, which are, however, narrower than those
of C. capilliformis. Fresh material that would match the holotype
is needed to confirm a systematic placement of this taxon.
Ceratostomella conica (Ellis & Everh.) M.E. Barr, Mycotaxon 46:
60. 1993.
Basionym:Ceratostoma conicum Ellis & Everh., Proc. Acad. nat.
Sci. Philad. 42: 226. 1890.
Specimen examined:USA, New Jersey, Newfield, on decaying wood of a pine
log, 1888 (holotype, NY).
Notes: The fungus is characterised by immersed to partially
erumpent ascomata with a rostrate ostiolum, cylindrical, short-
stipitate asci with a massive, non-amyloid apical annulus and
fusiform, hyaline 1‒3-septate, smooth-walled ascospores. It re-
sembles Chaetosphaeria, but differs in the morphology of the
apical annulus, which is relatively small and shallow in the latter
genus. In order to find a correct systematic placement for this
species, fresh material should be collected and DNA sequence
data generated.
Ceratostomella coprogena Massee, Bull. Misc. Inf., Kew: 105.
1913.
Specimen:Singapore, botanical garden, on animal dung, Burkill (holotype).
Notes: The type material could not be located at Kew (pers.
comm., B. Aguirre-Hudson). The coprophilous habitat is rather
atypical and it does not occur in any members of the Xen-
ospadicoidales or in other species described in Ceratostomella.
Ceratostomella cyclospora Kirschst., Verh. bot. Ver. Prov.
Brandenb. 48: 52. 1907.
Synonym:Calosphaeria cyclospora (Kirschst.) Petr., Annls
mycol. 22: 74. 1924.
Specimen:Germany, Grünauer forest near Rathenow, on decaying wood of
Pinus sp., 3 Jul. 1904, W. Kirschstein (holotype).
Notes: The holotype is apparently lost (pers. comm., E. Gerhardt,
B). According to the protologue (Kirschstein 1907) and a later
study of the holotype by Petrak (1924), the ascomata are rarely
solitary, mostly congregated in pairs of two or in a valsoid forma-
tion, partially or entirely immersed in bark or decorticated wood,
700‒900 μm diam with elongated necks, asci 30‒40 × 4‒6μm,
20‒25 μm long in the sporiferous part, cylindrical-clavate and
tapering at base, thick-walled and containing eight hyaline,
aseptate ascospores that are cylindrical, stronglycurved, 4 × 1 μm.
Petrak (1924) concluded that this species is better placed in
Calosphaeria and proposed a new combination in the latter genus.
The species fits the description of Barbatosphaeria hippocrepida
(R
eblov
a&
St
ep
anek 2009, R
eblov
aet al. 2015b), known only
from New Zealand, apart from the size of ascomata and asci. It is
R
EBLOV
AET AL.
56
likely that C. cyclospora represents a species closely related to
B. hippocrepida. Without studying the holotype or other repre-
sentative material we refrain from making any formal changes.
Ceratostomella debaryana (Auersw.) Sacc., Syll. fung. 1: 409.
1882.
Basionym:Gnomonia debaryana Auersw., Mycol. eur. Abbild.
S€
ammtl. Pilze Eur. 5–6: 23. 1869.
Specimen:Germany, Inselsberge, on decaying wood of Fagus sylvatica,
Fleischhak (holotype).
Notes: The type material of C. debaryana could not be located in
B (pers. comm., E. Gerhardt). According to the protologue and
illustration (Auerswald 1869), the ascomata are immersed,
globose, 140 μm diam with subcylindrical protruding necks, cy-
lindrical short-stipitate asci 68 × 6 μm and eight uniseriate,
ellipsoidal, hyaline, aseptate ascospores with granulose content.
The size of ascomata is too small to match the size of any genus
of Xenospadicoidales.
Ceratostomella echinata Ellis & Everh., N. Amer. Pyren. p. 195.
1892. (nom. nud., Art. 32.1).
Notes: Although Index Fungorum lists this name among species
of Ceratostomella, this species was not described; page 195 in
Ellis & Everhart (1892) refers to C. echinella (see above). No
type or authentic material is preserved in NY (pers. comm., B.M.
Thiers).
Ceratostomella excelsior Mouton, Bull. Soc. R. Bot. Belg. 36: 12.
1897.
Synonym:Endoxyla excelsior (Mouton) Munk, Bot. Tidsskr. 61:
66. 1965.
Specimen examined:Belgium, Beaufays near Li
ege, on decaying wood of
Fraxinus sp., V. Mouton (holotype, BR-93865-66).
Notes: The species bears a certain resemblance to Jattaea
based on the morphology of ascomata, ascospores and tapering
asci without a visible apical annulus and with a bulbose base
attached to ascogenous hyphae. However, it does not fit the
description of any known species of that genus (Damm et al.
2008, R
eblov
a 2011). Munk (1965) transferred the fungus to
the broadly perceived Endoxyla (Untereiner 1993). Represen-
tative material should be collected and subjected to phylogenetic
analysis in order to correctly classify this taxon.
Ceratostomella hystricina (Cooke) Sacc., Syll. fung. 2, Add. xxx.
1883.
Basionym:Ceratostoma hystricina Cooke, Grevillea 11: 109.
1883.
Specimen examined:USA, South Carolina, Aiken, on bark of Ficus sp., H.W.
Ravenel 2676 (syntypes, ex herb. M.C. Cooke, K 147286, K 147288); ibid., H.W.
Ravenel Fungi Amer. Exs. No. 674 (K 147290).
Notes: The protologue of C. hystricina and representative her-
barium material do not refer to the same fungus. Examination of
the three specimens from K cited above revealed a fungus that is
in agreement with observations of Ellis & Everhart (1892) and a
fungus distributed in Ellis & Everhart's N. Amer. Fungi. Exs. Ser.
II. No. 2349 (not seen). These specimens contain a fungus with
globose, immersed to superficial, densely aggregated ascomata
covered with a dark pink to brown crustose layer, with a cylin-
drical, upright, glabrous neck and minute, hyaline, aseptate,
allantoid ascospores 4‒4.5 × 1‒1.5 μm. Asci are present but
indistinct and visible merely as bundles of ascospores. However,
in the protologue, the fungus is described with ellipsoidal-
lanceolate ascospores 16‒18 × 6 μm(Cooke 1883) and the
same illustration is made on the envelope of the syntype (K
147286). Cooke based his description of C. hystricina on H.W.
Ravenel Fungi Amer. Exs. No. 674. The fungus present in the
examined collections is best placed in the Diaporthales; a spe-
cies of Valsa fide Ellis & Everhart (1892).
Ceratostomella leiocarpa Sacc. [as ‘lejocarpa’], Michelia 1: 370.
1878.
Specimen:Italy, Cansiglio, on decaying wood of Fagus sylvatica (holotype,
PAD).
Notes: The type material could not be located in PAD (pers.
comm., R. Marcucci). Based on characters of the ascomata, asci
and ascospores cited in the protologue by Saccardo (1878a), this
species fits best the description of Lentomitella cirrhosa.
Ceratostomella multirostrata (Fuckel) Sacc., Syll. fung. 1: 411.
1882.
Basionym:Ceratostoma multirostratum Fuckel, Jb. nassau. Ver.
Naturk. 23‒24: 129. 1870.
Specimen examined:Germany, Budenheimer Forest, on decaying wood of Pinus
sylvestris, spring (holotype, Fungi Rhen. Exs. No. 771, G).
Notes: The holotype contains empty ascomata that are superfi-
cial, confluent, with 1‒4 cylindrical upright necks and accom-
panied by a coelomycete forming stromatic conidiomata with
monostichous loculi. Based on the protologue (Fuckel 1870) and
illustration accompanying the type material, we cannot conclu-
sively attribute this species to any known genus.
Ceratostomella mycophila Rick, Brot
eria, s
er. bot. 5: 48. 1906.
Specimen:Brazil, Rio Grande do Sul, in hymenophore of Poria sp., 1905 (ho-
lotype, PACA 12733).
Notes: The type material is deposited in Herbarium Anchieta but
was not available to our study (pers. comm., M. S. Marchioretto,
PACA). According to the protologue (Rick 1906), the fungus fits
well the description of Ceratostomella s. str. It resembles
C. rostrata in habitat, size and morphology of cylindrical, pale
brown ascospores, but differs in shorter asci 25 × 6 μm vs. (26–)
30–39 × 5–6μminC. rostrata (R
eblov
a 2006). However, some
collections of C. rostrata can rarely have shorter asci, e.g.
Sclerom. Suec. Exs. No 116, PRM 666367 with asci
23–32 × 5–6μm. This collection is old, the ascus stipe is partly
disintegrated and asci appear slightly wrinkled and therefore
shorter. Recollection of fresh material of C. mycophila from its
original locality is recommended before it can be accepted in
Ceratostomella or confirmed as conspecific with C. rostrata.
Ceratostomella nyssicola (Berk. & M.A. Curtis) Sacc. [as ‘nys-
saecola’], Syll. fung. 1: 412. 1882.
Basionym:Sphaeria nyssicola Berk. & M.A. Curtis, [as ‘nys-
saecola’] Grevillea 4: 143. 1876.
Specimen examined:USA, Pennsylvania, wood of Nyssa sp., Michener 5166
(holotype, K 155069).
Notes: The holotype is in poor condition and apart from mostly
damaged ascomata, only a few pale brown, ellipsoidal and
slightly apiculate, aseptate, smooth-walled ascospores
9.5–10.5 × 4.5–5μm, which have a minute germ pore at each
end and a large drop inside, are present. Based on these
characters the fungus belongs in Xylomelasma (R
eblov
a 2006),
and it is likely an earlier name for X. sordida, the type species.
SORDARIOMYCETES
www.studiesinmycology.org 57
Ceratostomella rhenana (Auersw.) Sacc., Syll. fung. 1: 409.
1882.
Basionym:Gnomonia rhenana Auersw., Myc. Europ. Pyren.
5–6: 23. 1869.
Specimen examined:Germany, Nassau, on decaying wood, Fuckel ex Herb.
Barbey-Boissier No. 603 (K 84430, Fungi Rhen. Exs. No. 1804).
Notes: The type material of C. rhenana is apparently lost (pers.
comm., B. Hein, B). Winter (1887) based his description of
C. rhenana on a part of Fuckel's exsiccate collection (Fungi
Rhen. Exs. No. 1804, pro parte). Our revision of this specimen
from Fuckel's herbarium in G revealed Lentomitella vestita, but a
fungus sensu Auerswald (1869) and Winter (1887) was not
found. However, a specimen of Fungi Rhen. Exs. No. 1804
deposited in Kew (K 84430) contained the fungus, which is in
agreement with the protologue of Gnomonia rhenana (Auerswald
1869). The material in Kew is not suitable for typification due to
its poor condition and lack of asci. The systematic placement of
this fungus is unknown. Other parts of Fungi Rhen. Exs. No.
1804 need to be studied and fresh material of this fungus needs
to be recollected to investigate the relationship of C. rhenana with
Ceratosphaeria and other morphologically similar taxa. For a
detailed discussion on this species and revision of available
representative material see R
eblov
a (2009).
Ceratostomella rostrata var. levirostris Sacc., Syll. fung. 1: 408.
1882.
Notes: No type or authentic material of Ceratostomella rostrata
var. levirostris could be found in PAD (pers. comm., R. Marcucci).
Saccardo (1882) mentioned this taxon briefly in a discussion of
C. rostrata:‘Var. levirostris rostro non v. vix sulcato. Cum specie.’
Ceratostomella stricta (Pers.) Sacc., Syll. fung. 1: 410. 1882.
Basionym:Sphaeria stricta Pers., Syn. meth. fung. 1: 59. 1801.
Notes: The holotype could not be located in L (pers. comm., G.
Thijsse). Sphaeria stricta is distributed in Fries's Sclerom. Suec.
Exs. No. 148, but this collection was not examined. The fungus
was described with just a few words (Persoon 1801), and later
redescribed by Saccardo (1882) based on various materials from
European localities. Based on Saccardo's description of non-
stromatic densely aggregated, globose, glabrous ascomata with
a cylindrical neck, cylindrical-clavate asci with a slender stipe and
hyaline, subcylindrical, curved ascospores, the fungus would be
best placed in Calosphaeria or Jattaea in Calosphaeriales.
Ceratostomella stricta var. cingulata (Fr.) Sacc., Syll. fung. 12:
83. 1897.
Basionym:Sphaeria stricta var. cingulata Fr., Syst. mycol. 2: 474.
1823.
Notes: We could not locate any authentic material for this study.
Ceratostomella stricta var. majuscula Schulzer & Sacc., Hedwi-
gia 23: 42. 1884.
Synonym:Ceratostomella majuscula (Schulzer & Sacc.) Mussat,
in Saccardo, Syll. fung. 15: 84. 1901. (Nom. inval., Art. 36.1a, c)
Specimen:Hungary, Vinkovce, on decaying wood of Populus sp. (holotype).
Notes: Type material could not be located in W. The species is
described with hyaline ascospores 12–14 × 3 μm with 2–3
guttules (Schulzer & Saccardo 1884). The description is insuf-
ficient to transfer this fungus to any known genus. The fungus
was also illustrated in Schulzer (1869), cited in Saccardo &
Schulzer (1884) as Ill. Fung. Slav. no 845. In the absence of
the holotype, the illustration is the only surviving element, which
should serve as a lectotype. Schulzer's manuscript with illus-
trations is deposited at the Hungarian Academy in Budapest.
Unfortunately, the illustration could not be located (pers. comm.,
G. T
oth, Dept. of Manuscripts & Rare Books).
Ceratostomella stylophora (Berk. & Broome) Cooke, Grevillea
17: 49. 1889.
Basionym:Sphaeria stylophora Berk. & Broome, Ann. Mag. nat.
Hist., Ser. 3, 7: 453. 1861.
Specimen:UK, Scotland, Mossburnford, on bark of Acer platanoides, A. Jerdon
(holotype, K 84425).
Notes: The type material contained non-stromatic, ovoid ascomata
densely aggregated in small groups and erumpent through the thin
bark of a twig, with upright, cylindrical, partly flattened necks. The
asci were mostly disintegrated, only remnants of the upper half
with a distinct apical annulus could be seen. The ascospores are
hyaline, fusiform, slightly curved, 1-septate with short appendages
at each end. The fungus is better placed in the Diaporthales.
Ceratostomella subpilosa (Fuckel) Sacc., Syll. fung. 1: 411.
1882.
Basionym:Ceratostoma subpilosum Fuckel, Jb. nassau. Ver.
Naturk. 23–24: 128. 1870.
Specimen examined:Germany, Grünau, on decaying wood of Salix alba (ho-
lotype, Fungi Rhen. Exs. No. 2251, G).
Notes: The type material is overmature. The ascomata are
subglobose, glabrous with a fragile crumbling wall, immersed
with only necks emerging, ascospores 8–9×4–5μm, ellipsoidal
slightly tapering towards the ends, hyaline, aseptate, smooth-
walled, although some ascospores were observed to be verru-
culose. Given the poor condition of the specimen, the fungus
could not be properly identified.
Ceratostomella subsalsa (P. Crouan & H. Crouan) Sacc., Syll.
fung. 1: 412. 1882.
Basionym:Sphaeria subsalsa P. Crouan & H. Crouan, Florule
Finist
ere p. 25. 1867.
Specimen:France, Finist
ere, on decaying wood of Obione sp. (holotype).
Notes: The holotype material deposited in CO was not examined.
The identity and placement of this fungus is unknown; it was
described with globose, greenish ascomata 1–2 mm diam, thick-
walled clavate asci and ovoid, hyaline ascospores.
Ceratostomella unedonis Fabre, Annls Sci. Nat., Bot., S
er. 6. 15:
34. 1883.
Specimen:France, Vaucluse, on decaying wood of Arbutus unedo (holotype).
Notes: The holotype material deposited in FABR was not
examined. The identity and placement of this lignicolous fungus
is unclear. Based on the description of Saccardo (1883) the
ascomata are sparse, solitary, globose, ca. 330 μm diam, with an
upright central neck, short-stipitate asci 70–80 × 8 μm with eight
uniseriate, hyaline, aseptate, ovoid ascospores 8–10 × 5 μm.
The description is insufficient to identify this taxon.
ACKNOWLEDGEMENTS
This study was supported by the Project of the National Foundation of the Czech
Republic (GACR 506/12/0038, www.gacr.cz), by the MEYS under the project
CEITEC 2020 (LQ1601) and as a long-term research development project of the
Institute of Botany (No. RVO 67985939) and the Institute of Microbiology (No.
RVO 61388971) of the Czech Academy of Sciences. Computational resources
R
EBLOV
AET AL.
58
were provided by the CESNET LM2015042 and the CERIT ScientificCloud
LM2015085, under the programme “Projects of Large Research, Development,
and Innovations Infrastructures”.Thefield work in New Zealand (March–May
2005) was partly supported by Studienstiftung für mykologische Systematik und
€
Okologie. We are grateful to Walter Gams for the correction and grammatical
review of Greek and Latin epithets. We thank the curators of all cited herbaria
for their help in obtaining type and other herbarium material. Peter Johnston is
thanked for his assistance to M. R. in obtaining the Manaaki Whenua Fellowship
in 2005 and collecting permits for New Zealand. We thank Bevan Weir for his
assistance with obtaining strains of Lentomitella from the ICMP culture collec-
tion. We are grateful to Hermann Voglmayer for valuable discussion and sug-
gestions on phylogenetic analyses and Jacques Fournier, Sabine Huhndorf, and
Carol Shearer for valuable collections of Calyptosphaeria and Torrentispora.
Paul M. Kirk and Walter Gams are thanked for providing valuable background
for some nomenclatural problems.
APPENDIX A. SUPPLEMENTARY DATA
Supplementary data related to this article can be found at https://
doi.org/10.1016/j.simyco.2017.11.004.
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