Herzogia 36 (2), 2023: 283 –304 283

Octospora entosthodontophila,
a new smooth-spored bryophilous ascomycete
on Entosthodon spp.

Csaba Németh, Marcel Vega, Jorge Hernanz, Jan Eckstein & Lukáš Janošík

Abstract: Németh, C., Vega, M., Hernanz, J., Eckstein, J. & Janošík, L. 2023. Octospora entosthodontophila, a
new smooth-spored bryophilous ascomycete on Entosthodon spp. – Herzogia 36: 283 –304.
Octospora entosthodontophila, a new smooth-spored bryophilous ascomycete on Entosthodon spp., is described and
illustrated based on several collections from Hungary and Spain. The new species infects various species of the ter-
ricolous moss genus Entosthodon (Funariaceae). It is one of the first bryoparasitic Pezizales where molecular data
confirmed the broader host spectrum. The newly described species is characterized by orange apothecia with a con-
spicuous membranaceous slightly fimbriate margin, narrowly ellipsoid to subfusiform smooth ascospores with two
large and several small lipid bodies. A comparison with similar Octospora species and taxa infecting other members
of the moss family Funariaceae is also provided. A phylogenetic analysis using ITS, LSU, SSU, and EF1-α sequences
revealed that O. entosthodontophila forms a monophyletic group with O. excipulata, a taxon also infecting various
members of the Funariaceae.

Zusammenfassung: Németh, C., Vega, M., Hernanz, J., Eckstein, J. & Janošík, L. 2023. Octospora entosthodon-
tophila, eine neue bryophile Art auf Entosthodon spp. mit glatten Sporen. – Herzogia 36: 283 –304.
Octospora entosthodontophila, eine neue bryophile Art auf Entosthodon spp. mit glatten Sporen, wird anhand meh-
rerer spanischer und ungarischer Kollektionen beschrieben und dargestellt. Die neue Art infiziert verschiedene Arten
der erdbewohnenden Moosgattung Entosthodon (Funariaceae) und gehört damit zu den ersten moosparasitischen
Pezizales, bei denen ein breiteres Wirtsspektrum molekular bestätigt wurde. Sie ist durch orange Apothecien mit einem
deutlich membranartigen, leicht fimbriaten Rand, schmal-elliptische bis subfusiforme, glatte Ascosporen mit zwei
großen und mehreren kleinen Ölkörpern charakterisiert. Octospora entosthodontophila wird mit ähnlichen Octospora-
Arten sowie weiteren an Funariaceae parasitierenden Arten verglichen. Eine phylogenetische Untersuchung der Loci
ITS, LSU, SSU und EF1-α zeigt O. entosthodontophila in einer monophyletischen Gruppe mit O. excipulata, ein
Taxon, das ebenfalls verschiedene Mitglieder der Funariaceae infiziert.

Key words: bryoparasitic Pezizales, Funariaceae, host specificity, Pyronemataceae.

Introduction
The genus Octospora Hedw. s. l. (including all other related genera of bryophilous Pezizales)
has a worldwide distribution and contains ca. 200 described and numerous undescribed taxa.
All of them have a highly specialised bryophilous lifestyle, being closely associated with
various bryophyte hosts. They show a high morphological diversity in the shape and orna-
mentation of ascospores as well as in number, size, and pattern of lipid bodies inside the
ascospores (Janošík et al. 2023). Most of them possess ascospores ornamented with warts,
tubercles, ridges or reticulum and only around 30 % of the described species have smooth
284 Herzogia 36 (2), 2023

ascospores (Eckstein 2023). Ascospore ornamentation appears to be a relatively fast evolv-

ing trait. Species with smooth ascospores are scattered in multiple lineages throughout the
phylogeny of bryophilous Pezizales, and the ancestral state reconstruction of ornamentation
height showed that there were likely numerous transitions from ascospores with smooth sur-
face to strongly ornamented ascospores, as well as shifts in the opposite direction (Janošík
et al. 2023).
During a field trip in 2016 numerous apothecia of a suspected Octospora with smooth as-
cospores were found growing among shoots of Entosthodon fascicularis (Hedw.) Müll.Hal
(Funariaceae) in Hungary. In subsequent years, additional specimens with very similar mor-
phological features were collected in Hungary and Spain, all growing intimately associated
with various species of Entosthodon, a genus previously not reported as host of any bryophil-
ous ascomycete (Eckstein 2023).
Based on comparative microscopic and molecular studies, the above-mentioned collections
are described as Octospora entosthodontophila, a species new to science.

Material and methods

Sample collection and observation
The description of macroscopic and microscopic features is based on vital material collect-
ed in Hungary and Spain. Measurements were made in tap water. Absence of amyloidity of
asci was checked in Lugol’s solution (IKI). To determine quantitative ascospore parameters
at least 50 ascospores were measured from a spore print. The length, width, and Q value of
ascospores are presented in the following form: (lowest measured value) 5 % percentile value
‒ mean ‒ 95 % percentile value (highest measured value). Infection was studied in Lactic Acid
Cotton Blue (LACB). Scanning electron micrographs (SEM) were taken with a Zeiss EVO 40
SEM machine from air dried samples. Nuclei were visualized in apothecia upon staining with
DAPI (4',6-diamidino-2-phenylindole). A piece of hymenium was transferred to a 30 μl drop
of DAPI (2 μg/ml) on a microscope slide and heated at 90 °C for 1 min on a hot plate. After
at least 5 minutes at room temperature, the stained ascospores were observed using Olympus
BX51 (excitation 360 –370 nm and emission > 420 nm Olympus, Tokyo, Japan). The nomen-
clature of bryophytes follows Hodgetts et al. (2020). Coordinates are given in the WGS
84 format. The holotype voucher was deposited in the herbarium of the Hungarian Natural
History Museum (BP). Other studied specimens are housed in Herbarium of the University
of Alcalá de Henares (AH), Herbarium of the Botanic Garden and Botanical Museum Berlin-
Dahlem (B) and, BP, as well as in different private herbaria (LJ‒Lukáš Janošík, MAR‒Miguel
Ángel Ribes, CSN‒Csaba Németh).

DNA extraction, PCR amplification, and sequencing

DNA was extracted from fresh or dried apothecia using the Quick-DNA™ Fungal/Bacterial
Miniprep Kit (Zymo Research, Orange, USA). Sequence data were generated for four re-
gions: the large subunit of ribosomal DNA (LSU) was amplified with primers NL1 and NL4
(O’Donnell 1993) or LR5 (Vilgalys & Hester 1990), small subunit of rDNA (SSU) with
primers NS1 and NS6 (White et al. 1990), internal transcribed spacers (ITS) of rDNA (ITS1–
5.8S rDNA–ITS2 region) with primers ITS1F (Gardes & Bruns 1993) and ITS4 (White
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 285

et al. 1990), and the elongation factor 1-alpha (EF1-α) with primers EF1–983F (Rehner &
Buckley 2005) and EF1-octoR (Vega et al. 2021b). The PCR products were purified with
Agencourt AMPure XP beads (Beckman Coulter, Massachusetts, USA). Both strands of the
PCR fragments were then sequenced with the primers used for amplification at the Sequencing
Laboratory of the OMICS Core Facility, BIOCEV (Vestec, Czech Republic). Obtained se-
quences were checked and edited in Geneious 9.1 (Biomatters, Auckland, New Zealand) and
deposited in GenBank.

Phylogenetic analysis
Specimens used in the analysis and their GenBank accession numbers are listed in Table 1,
with newly obtained sequences in bold letters. Newly obtained sequences of LSU, SSU, ITS
and EF1-α were used together with other sequences of bryophilous Pezizales from previous
studies (Stenroos et al. 2010, Lindemann et al. 2014, Vega et al. 2017, Egertová et al.
2018a, Egertová et al. 2018b, Sochorová et al. 2019, Vega et al. 2019, Sochorová et al.
2020, Eckstein et al. 2021, Sochorová et al. 2021, Vega et al. 2021b, Eckstein et al. 2022,
Janošík et al. 2023), as well as Otidea concinna (Pers.) Sacc. (Schoch et al. 2012, Hansen
& Olariaga 2015), serving as an outgroup. The sequences were aligned with MAFFT (on-
line version 7) using the E-INS settings (Katoh et al. 2019). The ambiguously aligned parts
of the ITS region were removed with BMGE (Criscuolo & Gribaldo 2010) and the most
suitable substitution model for each region of the concatenated dataset was determined in
PartitionFinder 2.1.1 (Lanfear et al. 2017), using the BIC and a greedy search, which selected
the general time reversible substitution model (GTR + G + I) as the best fitting for all parti-
tions except ITS region, for which Hasegawa-Kishino-Yano substitution model (HKY + G + I)
was used. Bayesian analysis (BI) was conducted using MrBayes 3.2.3 (Ronquist et al. 2012),
with two independent runs of five million generations and four chains, sampling every 1000th
generation, the first 25 % of samples were discarded as burn-in. Maximum likelihood analysis
(ML) was performed using raxmlGUI 2.0 (Edler et al. 2021, Stamatakis 2014) and analysed
as a partitioned dataset under the GTRCAT model with 1000 bootstrap iterations. Trees based
on the analysis of individual regions, using the same parameters as the concatenated dataset,
are shown in Supplementary Fig. 1.

Results
Phylogenetic analysis
Based on the molecular markers used, O. entosthodontophila forms a distinct and well sup-
ported clade together with O. excipulata (Clem.) Benkert (a smooth-spored species infect-
ing the different moss species Funaria hygrometrica Hedw., Physcomitrium patens (Hedw.)
Mitt., and Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid., belonging also to the
family Funariaceae (Benkert 2007, Eckstein & Eckstein 2013). Other smooth-spored spe-
cies with similar ascospore morphology (i.e. O. axillaris (Nees) M.M.Moser, O. axillaris var.
tetraspora Benkert, O. leucoloma Hedw., O. leucoloma var. tetraspora (Fuckel) Benkert, O.
gemmicola Benkert agg., O. gemmicola var. tetraspora Benkert, O. coccinea (P.Crouan &
H.Crouan) Brumm. agg., O. coccinea var. tetraspora Benkert, O. humosa (Fr.) Dennis, O. itze-
rottii Benkert, O. musci-muralis Graddon, O. musci-muralis var. neglecta (Dennis & Itzerott)
Benkert) have a different position in the tree (Fig. 1).
Table 1. Collections used in the phylogenetic analysis with voucher information and GenBank accession numbers. Newly generated sequences are in bold.
286

Species Herbarium code Country, collector Host GenBank accession numbers

LSU SSU ITS EF1-α
Lamprospora dictydiola PRM 945794 Czech Republic, Z.Egertová Tortula muralis MF754056 MK569365 MH818434 MF754054
Lamprospora gibbosa B 70 0100017 (holotype) France, M. Vega Fissidens crassipes MT792691 MT792712 MT783997
Lamprospora miniata PRC 4122 Slovakia, L. Janošík Tortula protobryoides MH818444 MH818430
Lamprospora rehmii S F317032 (epitype) Spain, R. Martínez-Gil Pleuridium acuminatum MH087070 MT792719 MH087068
Lamprospora sylvatica PRM 946415 (holotype) Ukraine, Z. Egertová & Dicranum montanum MG947604 MK569367 MG947607 MK569290
M. Sochor
Neottiella albocincta PRM 945796 Slovakia, P. Včelička Atrichum undulatum MF754059
Neottiella rutilans B 70 0100473 Poland, J. Eckstein Oligotrichum hercynicum MK569313 MK569336 MK569288
Neottiella vivida PRM 945797 Czech Republic, Z.Egertová Polytrichum piliferum MF066068 MK569337 MF066095 MF754051
Octospora affinis PRM 945798 Czech Republic, Lewinskya affinis MF754075 MK569347 MF754045
A.Polhorský,L. Janošík &
Z.Egertová
Octospora axillaris PRC 4940 Czech Republic, L.Janošík Tortula acaulon ON087108 ON087186 ON093877
Octospora axillaris PRM 954016 Czech Republic, Z.Egertová Tortula acaulon MW242829 MW242828 MW430761
Octospora axillaris var. PRC 4941 Slovakia, L. Janošík Tortula acaulon ON087109 ON087187 ON093878
tetraspora
Octospora bridei PRM 935151 Czech Republic, Z.Egertová Ephemerum serratum MF754061 MT001890
Octospora bryi-argentei B 70 0005999 (holotype) Germany, D. Benkert Bryum argenteum MZ343192
Octospora coccinea agg. PRC 4942 Austria, L. Janošík Bryum cf. klinggraeffii ON087110
Octospora coccinea agg. PRC 4974 Czech Republic, L.Janošík Encalypta vulgaris ON087144
Octospora coccinea var. JE58964 Germany, J. Eckstein Bryum subapiculatum OR036966
tetraspora
Octospora conidiophora PRM 951743 South Africa, Trichosteleum perchlo- MK569321 MK569351 MK569297
(holotype) Z.Egertová & M.Sochor rosum
Octospora doebbeleri PRM 954007 (holotype) Czech Republic, Dicranoweisia cirrata MW152148 MW152156 MW159137
Z.Sochorová & M.Sochor
Octospora entosthodon- BP 112291 Spain, M. Vega, Entosthodon attenuatus OR036967 OQ930002
tophila M. Á. Ribes & S. Tello
Herzogia 36 (2), 2023
Species Herbarium code Country, collector Host GenBank accession numbers
LSU SSU ITS EF1-α
Octospora entosthodon- PRC 4986 Spain, Canary Islands, Entosthodon attenuatus ON087089 ON087172 OQ930003
tophila Tenerife, L.Janošík
Octospora entosthodon- BP 112289 (holotype) Hungary, C. Németh Entosthodon fascicularis OR036969 OQ930009 OQ930004 OQ928399
tophila
Octospora entosthodon- BP 112286 Hungary, C. Németh Entosthodon cf. fascicu- OR036970
tophila laris
Octospora entosthodon- BP 112292 Spain, J. Hernanz Entosthodon schimperi OR036971 OQ930010 OQ930005 OQ928400
tophila
Octospora entosthodon- BP 112290 Spain, Canary Islands, Entosthodon attenuatus OR036972 OQ930011 OQ930006
tophila Tenerife, M.Vega
Octospora entosthodon- BP 112287 Spain, J. Hernanz Entosthodon schimperi OR036973 OQ930012 OQ930007 OQ928401
tophila
Octospora entosthodon- BP 112288 Spain, J. Hernanz Entosthodon fascicularis OR036974 OQ930013 OQ930008 OQ928402
tophila
Octospora excipulata PRM 945800 Czech Republic, Z.Egertová Funaria hygrometrica MF754062 MK569369 MF754047
Octospora excipulata B 70 0108146 France, M. Vega Funaria hygrometrica OR036975
Octospora gemmicola B 70 0010006 (holotype) Germany, S. Rätzel Bryum atrovirens OL832140
Octospora gemmicola var. B 70 010007 (holotype) Germany, D. Benkert Bryum klinggraeffii OL832141
tetraspora
Octospora gyalectoides PRC 4694 Czech Republic, L.Janošík Tortula acaulon MZ343187 MZ343177 MZ336036
Octospora gyalectoides PRC 4949 Czech Republic, L.Janošík Tortula acaulon ON087117 ON087193 ON093883
Octospora humosa s.str. PRM 945802 Czech Republic, Z.Egertová Polytrichum piliferum MF754074 MK569343 MF754043
Octospora itzerottii PRC 4951 Czech Republic, L.Janošík Pterygoneurum ovatum ON087119
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete

Octospora itzerottii BP 112285 Hungary, C. Németh Pterygoneurum subses- OR036976

sile
Octospora kelabitiana PRM 945781 Malaysia, Z. Egertová & Riccardia sp. MF754065 MK569372 MF754048
M. Sochor
Octospora leucoloma PRM 945804 Czech Republic, Z.Egertová Bryum argenteum MF754063 MK569370 MF066093
Octospora leucoloma PRC 4952 Czech Republic, L.Janošík Bryum argenteum ON087120 ON087195 ON093885
287
Species Herbarium code Country, collector Host GenBank accession numbers
288

LSU SSU ITS EF1-α

Octospora leucoloma var. PRC 4953 Austria, L. Janošík Bryum argenteum ON087121 ON087196 ON093886
tetraspora
Octospora meslinii PRM 954637 (epitype) Hungary, C. Németh Grimmia pulvinata MW152147 MW152158 MW159139
Octospora musci-muralis PRC 4955 Czech Republic, L.Janošík Grimmia pulvinata ON087123 ON087198 ON093888
Octospora musci-muralis PRC 4991 Slovakia, L. Janošík Schistidium crassipilum ON087096
var. neglecta
Octospora neerlandica PRC 4691 Germany, M. Vega & Syntrichia ruralis agg. MZ343185 MZ343176 MZ336035
T. Richter
Octospora cf. orthotrichi CNF 2/10561 Croatia, Z. Egertová & Orthotrichum diaphanum MK569314 MK569342 MK569311
M. Sochor
Octospora pseudoampez- PRM 935156 Czech Republic, Schistidium crassipilum MF754069 MK569339 MF754050
zana Z. Egertová & M.Sochor
Octospora rubens agg. PRM 954641 Spain, M. Vega Ceratodon purpureus MW221931 MW206790 MW219144
Octospora rustica agg. PRC 4688 Slovakia, L. Janošík Ceratodon purpureus MZ343182
Octospora rustica agg. PRC 4690 Czech Republic, L. Janošík Ceratodon purpureus MZ343184 MZ336034
& K. Daňková
Octospora wrightii PRC 4617 Czech Republic, L.Janošík Amblystegium serpens MN994534 MN994517 ON084928 MN990994
Octosporella junger- TUR 178050 Switzerland, P. Döbbeler Plagiochila asplenioides EU940133 EU940060
manniarum
Octosporella perforata PRM 945808 Czech Republic, Z.Egertová Porella platyphylla MF754060 MK569368 MF754052
Octosporopsis erinacea PRM 945774 (isotype) Malaysia, Z. Egertová & Dumortiera hirsuta MF754057 MK569338 MF754041
M. Sochor
Octosporopsis nicolai UL151–13 Germany, M. Vega Lunularia cruciata KF771033 KF771042
Otidea concinna KH.09.183 (S) (epitype) Sweden, K. Hansen & - NG_060279 NG_064990 NR_172221 KM823275
I. Olariaga
Herzogia 36 (2), 2023
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 289

0.98/97 Octospora leucoloma PRM 945804

1/78
Octospora leucoloma PRC 4952
Octospora leucoloma var. tetraspora PRC 4953
1/93
1/97 Octospora coccinea agg. PRC 4942
Octospora coccinea var. tetraspora JE58964
0.91/73
Octospora gemmicola var. tetraspora B 70 0010007
Octospora gemmicola B 70 0010006
1/100 Octospora axillaris PRC 4940
1/99
Octospora axillaris PRM 954016
1/100 Octospora gyalectoides PRC 4694
0.99/91
Octospora gyalectoides PRC 4949
Octospora itzerottii PRC 4951
1/99
Octospora itzerottii BP 112285
Octospora axillaris var. tetraspora PRC 4941
0.99/61 Octospora entosthodontophila BP 112290
0.99/81 Octospora entosthodontophila BP 112287
Octospora entosthodontophila BP 112291
Octospora entosthodontophila PRC 4986
1/100
Octospora entosthodontophila BP 112289 HOLOTYPE
Octospora entosthodontophila BP 112292
1/100 0.95/79 Octospora entosthodontophila BP 112288
Octospora entosthodontophila BP 112286
1/100 Octospora excipulata PRM 945800
0.92/92 0.75/66
Octospora excipulata MV190610-02
Octospora coccinea agg. PRC 4974
0.98/59 Octospora bridei PRM 935151
Octospora rustica agg. PRC 4688
Neottiella vivida PRM 945797
1/100
Neottiella rutilans B 70 0100473
Neottiella albocincta PRM 945796
1/100 Octosporopsis nicolai UL151-13
Octosporopsis erinacea PRM 945774
0.99/77 Octosporella perforata PRM 945808
1/85
Octosporella jungermanniarum TUR 178050
Lamprospora gibbosa B 70 0100017
0.69/17
Octospora rubens agg. PRM 954641
0.93/55
1/100 Lamprospora miniata PRC 4122
1/100
Lamprospora dictydiola PRM 945794
1/82 1/100
Lamprospora sylvatica PRM 946415
Lamprospora rehmii S F317032
1/84
Octospora doebbeleri PRM 954007
0.81/70 Octospora rustica agg. PRC 4690
1/99
Octospora neerlandica PRC 4691
1/85 Octospora bryi-argentei B 70 0005999
0.95/87 Octospora humosa agg. PRM 945802
1/90
Octospora wrightii PRC 4617
1/100
Octospora affinis PRM 945798
1/100 Octospora cf. orthotrichi CNF 2/10561
1/100 Octospora kelabitiana PRM 945781
Octospora conidiophora PRM 951743
1/100 Octospora pseudoampezzana PRM 935156
Octospora meslinii PRM 954637
0.67/- 1/100 Octospora musci-muralis PRC 4955
Octospora musci-muralis var. neglecta PRC 4991
Otidea concinna KH.09.183 (S)

Fig. 1. Fifty percent majority rule Bayesian phylogram obtained from the concatenated LSU, SSU, and EF1-α se-
quences showing the phylogenetic relationship of newly described Octospora entosthodontophila with other bryophil-
ous Pezizales. Numbers above branches represent Bayesian posterior probability scores and RAxML bootstrap sup-
port values, respectively. GenBank accession numbers and additional collection information are indicated in Table 1.

Haplotype networks

We detected little polymorphism only within LSU and ITS among individual collections.
In LSU three different haplotypes were observed, five collections were fully identical (BP
112289 – holotype, PRC 4986, BP 112288, BP 112292, and BP 112286), two others differed
only in a single nucleotide (BP 112287 and BP 112290) and one other in another single nucleo-
tide (BP 112291). In ITS four different haplotypes were found, four collections proved to be
fully identical (PRC 4986, BP 112288, BP 112292, and BP 112291). BP 112289 – holotype, BP
112287 and BP 112290 were represented with different haplotypes differing only in a single
290 Herzogia 36 (2), 2023

Fig. 2. Haplotype networks for the collections of Octospora entosthodontophila showing all variability in the ITS and
LSU as well as the observed host pattern and distribution of sequenced collections. Individual circles depict unique
haplotypes and the diameter of each circle is proportional to the number of collections it represents. Nucleotide dif-
ferences are denoted by the hatch marks across the lines connecting haplotypes with each mark representing a single
nucleotide difference.

indel. In SSU and EF1-α we have not found any polymorphism at all among the sequenced
collections. Interestingly, we did not observe any connection between the haplotype patterns
and the host species or geographical area (Fig. 2).

Description
Octospora entosthodontophila C.Németh, M.Vega, Janošík & J.Hernanz sp. nov. (Figs. 3 – 6) [MycoBank
no. MB 848858]
Etymology: entosthodon = referring to the host genus Entosthodon. Phila means ‘attraction or affinity
to something’.
Diagnosis: Octospora entosthodontophila differs from other species of Octospora by narrowly el-
lipsoid to subfusiform ascospores with a smooth surface as well as two large (one of which always
somewhat smaller) and several small lipid bodies inside, together with an infection on stem and basal
leaf cells of various species of the moss genus Entosthodon.
Holotype: Hungary, Vértes Mts, Csákvár, Mt Szóló-kő, 47°23'05.0''N/18°26'05.3''E, 235 m alt., on slope steppe
on rocky soils, 5 March 2016, leg. C. Németh. Host: Entosthodon fascicularis, BP 112289. GenBank: OQ930004
(ITS), OR036969 (LSU), OQ930009 (SSU), OQ928399 (EF1-α).
Isotype: CSN 7778.
Hosts: Entosthodon attenuatus (Dicks.) Bryhn, E. fascicularis, and E. schimperi Brugués.
Macroscopic features (Fig. 3): Apothecia scattered or gregarious, directly growing on shoots of the
hosts or on soil nearby, young spherical, in mature state discoid, sessile, (0.5)1‒4 mm in diameter,
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 291

A B C

D E F G

H I

Fig. 3. Octospora entosthodontophila, macroscopic features (D BP 112289 ‒ holotype, E BP 112291, F BP 112290,

G BP 112287, H‒I CSN 10248). A – Habitat in Vértes Mts, Hungary. B ‒ Habitat in Cenicientos, Spain. C – Habitat
in El Rosario, Tenerife, Spain. D–I ‒ Apothecia with the host species, D, H–I ‒ Entosthodon fascicularis, E‒F ‒
Entosthodon attenuatus, G ‒ Entosthodon schimperi. Photos A, D, G‒I C. Németh, B J. Hernanz, C F M. Vega, E M.
Á. Ribes.

mostly with a conspicuous membranaceous, slightly fimbriate margin when mature, up to 0.3 mm
wide. Hymenium yellow-orange or bright-orange, margin paler than the hymenium.
Microscopic features (Fig. 4, 5): Asci cylindrical, 245‒350 × 20‒24 µm, 8-spored, opercula-
te, inamyloid, pars sporifera 100‒150 µm, bifurcate at the base, arising from perforated croziers.
Paraphyses filiform, mostly curved, rarely straight, pluriseptate, sometimes branched, last cell api-
cally inflated containing orange carotenoid pigment, terminal cell 40 – 80 × (4 –)5 –10(–12) µm, ba-
292 Herzogia 36 (2), 2023

A B C

E
Fig. 4. Octospora entosthodontophila, microscopic features (A‒B BP 112286, C CSN 10443, D CSN 10248, E BP
112287). A ‒ Upper part of paraphyses in water. B ‒ Paraphyses and ascospores inside asci in water. C ‒ Cross section
of an apothecium showing subhymenium, medullary, and ectal excipulum. D‒E ‒ Ascospores in ascus in water. Scale
bars: A‒E = 20 µm. Photos A‒B, D‒E C. Németh, C J. Hernanz

sally only 2 µm wide, no VBs seen. Ascospores uniseriately to subbiseriately arranged within the
ascus, smooth, uninucleate, narrowly (elongated) ellipsoid to subfusiform, sometimes slightly asym-
metric, (24.0 –)25.0 –25.6 –26.3(–27.6) µm long and (10.0 –)10.4 –10.7–11.3(–11.6) µm wide, Q=2.2–
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 293

A B

C D

Fig. 5. Octospora entosthodontophila, A‒B ‒ SEM pho-

tos of ascospores (A‒B BP 112289 ‒ holotype). C‒E
Fluorescence photos of uninucleate ascospores stained
with DAPI (C‒E BP 112291). Scale bars: A‒B = 2 µm,
E C‒E = 10 µm. Photos A‒B L. Szabó, C‒E L. Janošík.

2.3 –2.4(–2.6) [only free ascospores considered], with two large, slightly unequal lipid bodies, the
larger (7.4 –)7.8 – 8.2– 8.6(–9.2) µm and the smaller (6.1–)6.7–7.2–7.5(– 8.1) µm in diameter accompa-
nied by several smaller ones, diameter (0.9 –)1.3 –1.6 –2.1(–2.9) µm.
Medullary excipulum about 60 –120 µm thick comprising variously shaped cells forming a textura intri-
cata-angularis. Ectal excipulum 70 –190 µm thick, composed of textura globulosa-angularis. Margin
is composed of two layers: inner layer ca. 80 µm thick, extending from the ectal excipulum and formed
by textura globulosa-angularis, the outer layer ca. 80 µm thick, formed by textura prismatica.
Infection (Fig. 6): Hyphae colourless, smooth, thick walled, irregularly growing on the surface of stem
and basal leaf cells of the host species (Entosthodon attenuatus, E. fascicularis, and E. schimperi),
with ramifications and anastomoses, 2.5 – 6(– 8) µm wide, hyphae concentrating on basal stem parts
(rhizoid zone) and over young stems or bud-like stem innovations, sometimes within perigonia, in
heavy infections forming a thin coating around the stem or around stem innovations, coating consisting
of densely packed hyphal cells containing few appressoria, no hyphal relations to rhizoids observed.
Appressoria seen from above elliptical to broadly elliptical to almost round, apically sometimes exten-
294 Herzogia 36 (2), 2023

Fig. 6. Octospora entosthodontophila, drawings of infection structures on basal stem parts and basal leaf cells of
Entosthodon schimperi showing the one-celled terminal appressoria (BP 112287). Scale bar = 20 µm. Drawings J.
Eckstein.

ded, (in CB) 16 –25 µm long, 10 –16(–22) µm wide, 1-celled or with a thin septum, terminal, bryophyte
cell wall around the starting point of the haustorium thickened and forming an elliptical structure,
appressoria within a hyphal tissue may lose their typical shape. Haustoria usually one per host cell, as
a ramified, coiled filament, difficult to observe.
Additional specimens examined:
Hungary, Vértes Mts, Csákvár, Mt Szóló-kő, 47°23'03.8''N/18°26'04.6''E, 230 m alt., on a slope steppe on rocky
soils, 13 March 2016, leg. C. Németh. Host: Entosthodon fascicularis, accompanying bryophytes: Bryum dichoto-
mum Hedw., Pleuridium sp., Riccia ciliata Hoffm., Weissia sp., (CSN 7862), ibid. 8 March 2017 (CSN 8538), 24
Nov 2020 (CSN 10248), 6 March 2021 (CSN 10470).
Hungary, Gerecse Mts, Süttő, Mt Nagy-Teke, 47°42'00.3''N/18°26'32.9''E, 338 m alt., in dry grassland on limesto-
ne bedrock, 18 Nov 2020, leg. C. Németh, P. Finy & A. Koszka. Host: Entosthodon cf. fascicularis, accompanying
bryophytes: Bryum dichotomum Hedw., Pleuridium sp., Riccia ciliifera Link (BP 112286).
Spain, Canary Islands, Tenerife, Igueste de San Andrés, Lomo de las Casillas, 28°33'9.1''N/16°9'27.4''W, 545 m alt.,
trail margin at the edge of a humid forest, 19 Jan 2017, leg. L. Janošík. Host: Entosthodon attenuatus, accompany-
ing bryophytes: Gongylanthus ericetorum (Raddi) Nees, Phymatoceros bulbiculosus (Brot.) Stotler, W.T.Doyle &
Crand.-Stotl., Trichostomum sp. (PRC 4986).
Spain, Canary Islands, Tenerife, El Rosario, Camino Madroño Goteras, 28°26'25.5''N/16°23'08.4''W, 1140 m alt.,
on banks beside the forest road, 5 Jan 2021, leg. M. Vega. Host: Entosthodon attenuatus, accompanying bryophy-
tes: Gongylanthus ericetorum, Weissia sp. (BP 112290, CSN 10447).
Spain, Canary Islands, Tenerife, El Rosario, above the Barranco Gavilanes, 28°26'11.8''N/16°23'00.8''W, 1131 m
alt., beside a forest road, 23 Dec 2022, leg. M. Vega, M. Á. Ribes & D. Chavez. Host: Entosthodon attenuatus,
accompanying bryophytes: Gongylanthus ericetorum, Fissidens sp. (CSN 11377).
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 295

Spain, Canary Islands, Tenerife, San Juan de la Rambla, Área Recreativa La Tahona, 28°21'02.1''N/16°37'45.1''W,
1126 m alt., on a slope above a path, 19 Dec 2022, leg. M. Vega, M. Á. Ribes, R. Negrín & D. Chavez. Host:
Entosthodon attenuatus, accompanying bryophyte: Gongylanthus ericetorum (CSN 11376).
Spain, Andalusia, Jaén, Santa Elena, near the road N-Iva, 38°20'50''N/3°31'46.00''W, 730 m alt., on mountain slope, 22
Feb 2020, leg. M. Vega, M. Á. Ribes & S. Tello. Host: Entosthodon attenuatus, accompanying bryophytes: Fissidens
sp., Gongylanthus ericetorum, Fossombronia sp., Weissia sp. (BP 112291, MAR 220220 573, CSN 10445).
Spain, Madrid, Cenicientos, Las Tejoneras, 40º16'1.79''N/4º32'41.95''W, 690 m alt., in an abandoned old fruit orchard
on granitic sediments, 2 Jan 2021, leg. J. Hernanz. Host: Entosthodon schimperi, accompanying bryophytes: Fissidens
sp., Riccia sp., Bryum sp., Fossombronia sp. (BP 112292, CSN 10443), ibid. 13 Feb 2021 (AH-56306, CSN 10474).
Spain, Madrid, San Martín de Valdeiglesias, near the road N-501, 40º21'23.98''N/4º23'44.48''W, 650 m alt., in an
abandoned field of almond trees on granitic sediments, 13 Feb 2021, leg. J. Hernanz. Host: Entosthodon schimperi,
accompanying bryophytes: Fissidens sp. (AH-56307, BP 112287).
Spain, Madrid, Rozas de Puertoreal, Arroyo de los Morales, 40º18'46.84''N/4º31'8.65''W, 780 m alt., at a wet glade in
a riparian forest with Alnus and Salix on acid shale soil, 17 Feb 2021, leg. J. Hernanz. Host: Entosthodon fascicularis,
accompanying bryophytes: Bryum dichotomum Hedw., Tortula truncata (Hedw.) Mitt. (AH-56308, BP 112288).

Discussion

The vast majority of bryophilous Pezizales have extremely high host specificity, with a sin-
gle bryophyte species as host (Németh 2017, Eckstein et al. 2021, Sochorová et al 2021).
Reported multiple distantly related host species usually hint to an unrevealed taxonomic di-
versity (Vega et al. 2017, 2019). On the other hand, a single bryophyte species can host mul-
tiple unrelated, but morphologically almost indistinguishable fungal species, as was found in
Lamprospora tuberculata Seaver agg. on Pleuridium subulatum (Hedw.) Rabenh. (Vega et al.
2021b) and Octospora rustica (Velen.) J.Moravec agg. on Ceratodon purpureus (Hedw.) Brid.
(Eckstein et al. 2021). Occasionally, however, a true broader host specificity can be observed,
represented with closely related host species belonging usually to the same genus (Benkert
1998, Eckstein et al. 2022) and rarely to different genera (Sochorová et al. 2019).
In Octospora entosthodontophila three species of Entosthodon have hitherto been observed
as host. Entosthodon attenuatus is a Mediterranean-Atlantic species growing usually on per-
sistently moist soils of steep banks, ditches, earth-covered boulders and rock ledges, or rocky
slopes (Blockeel 2014). Entosthodon fascicularis is a widely distributed Temperate species
predominantly with North American–western Eurasian distribution, preferring recently dis-
turbed, seasonally wet soil of calcareous grasslands, arable fields, and several other habitats
(Blockeel 2014). Entosthodon schimperi is a species with Mediterranean distribution grow-
ing on dry and exposed soils (Casas et al. 2020, Hodgetts & Lockhart 2020).
Octospora entosthodontophila is known so far only from some localities in Hungary and Spain
(Fig. 2), occurring in significantly different habitats, according to the different ecology of its
host species. The two Hungarian localities are situated in open dolomite and limestone rocky
grasslands (Fig. 3A). The localities in Central Spain represent abandoned old fruit orchards
among Quercus ilex L., Q. suber L., and Prunus dulcis (Mill.) D.A.Webb trees on acidic
sandy soil overlying granitic sediments (Fig. 3B), as well as a wet glade of a riparian forest.
In Andalusia O. entosthodontophila could be detected under plants of Cistus sp. and other
shrubs growing beside a dried-out rivulet which only carries water in winter after heavy rain,
whereas the Tenerife collections all grew on banks besides forest roads or trails. Interestingly
the locality near El Rosario (Fig. 3C) is only 350 metres away from the type locality of the
recently described Lamprospora angularis M.Vega, Ribes & Janošík. A detailed description
of the ecology of the type locality of L. angularis can be found in Vega et al. (2021a).
296 Herzogia 36 (2), 2023

A B C

D E F

G H I

J K L

M N O
Fig. 7. Apothecia of similar species of bryophilous Pezizales with smooth
ascospores and more than one lipid body between shoots of their hosts.
A ‒ Octospora entosthodontophila with Entosthodon fascicularis (CSN
10248), B ‒ O. gemmicola var. tetraspora with Bryum sp. (CSN 9599),
C ‒ O. axillaris var. tetraspora with Tortula acaulon (CSN 9797), D ‒ O.
coccinea var. coccinea with Bryum sp. (CSN 10773), E ‒ O. coccinea var.
P tetraspora with Bryum sp. (CSN 10269), F ‒ O. gemmicola var. gemmi-
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 297

Octospora entosthodontophila is the only species of bryophilous Pezizales known to infect

species of the genus Entosthodon. Further features based on which O. entosthodontophila
can easily be identified and distinguished from the similar congeners are the shape and size
of the ascospores, as well as the number, size, and arrangement of the lipid bodies inside the
ascospores. Characteristically, two large but slightly unequal lipid bodies are present in mature
ascospores, arranged almost symmetrically and accompanied by several smaller ones.

Comparison with other species of Octospora with a similar ascospore morphology (Figs.
7– 8, Table 2)
Ascospores with a smooth surface and more than one lipid body inside are considered to be
similar to O. entosthodontophila. Data, measurements and information concerning the host
bryophytes are taken from Eckstein (2023).
Octospora gemmicola var. tetraspora has ascospores that are (21–)22–26(–30) × (9 –)9.5 –10.5(–
13) µm with an ellipsoid to narrowly ellipsoid shape and usually containing two or three larger
lipid bodies accompanied by several small ones. Octospora gemmicola var. tetraspora differs
from O. entosthodontophila in having 4-spored asci, a different size of the two larger lipid bod-
ies (8 –10 µm and 5 –7 µm vs. (7.4 –)7.8 – 8.2– 8.6(–9.2) µm and (6.1–)6.7–7.2–7.5(– 8.1) µm in
diameter, respectively), as well as host preferences of Bryum spp. (Fig. 7B, 8B).
Octospora axillaris var. tetraspora is characterized by narrowly ellipsoid-subfusiform as-
cospores, (22–)26 –32(– 40) × (9 –)10 –12(–15) µm, containing mostly three larger lipid bodies,
the largest one (10 –)10.5 –11.5(–12) µm, situated in the middle encompassed by two smaller
ones (4.5 –)5.5 –7.5(– 8) µm and accompanied by several small lipid bodies. It differs from O.
entosthodontophila in having 4-spored asci, larger ascospores, a different arrangement of lipid
bodies and a different host: Tortula acaulon (With.) R.H.Zander (Fig. 7C, 8C).
Octospora coccinea var. coccinea agg. has narrowly ellipsoid to narrowly fusiform ascospores,
(20 –)24 –30(–33) × (7–)8 –10(–11) µm, with four lipid bodies, two larger ones in the center
and two smaller ones near the ends. Octospora coccinea var. coccinea differs from O. ento-
sthodontophila in having larger and narrower ascospores, a different arrangement of lipid bod-
ies, as well as alternative host preferences, including Bryum spp., Encalypta vulgaris Hedw.
and Acaulon muticum (Hedw.) Müll.Hal. (Fig. 7D, 8D).
Octospora coccinea var. tetraspora is characterized by 4-spored asci with narrowly ellipsoid
to fusiform ascospores, (24 –)30 –34(– 40) × (8 –)9 –12(–15) µm, with four lipid bodies, two
larger ones in the center and two smaller ones near the ends. Octospora coccinea var. tetraspo-
ra differs from O. entosthodontophila in having 4-spored asci, considerably larger ascospores,
a different arrangement of lipid bodies and Bryum spp. as host bryophytes (Fig. 7E, 8E).

cola with Bryum sp. (CSN 9815), G ‒ O. axillaris var. axillaris with Tortula acaulon (CSN 10340), H ‒ O. musci-
muralis var. muralis with Grimmia pulvinata (CSN 10225), I ‒ O. musci-muralis var. neglecta with Schistidium
crassipilum (CSN 10803), J ‒ O. nemoralis with Fissidens viridulus (MV20140228–05), K ‒ O. guestfaliensis with
Tortella tortuosa (LJ 21014), L ‒ O. leucoloma var. leucoloma with Bryum argenteum (CSN 9051), M ‒ O. leucoloma
var. tetraspora with Bryum argenteum (CSN 10180), N ‒ O. humosa with Polytrichum piliferum (CSN 10282), O ‒ O.
itzerottii with Pterygoneurum ovatum (CSN 9814), P ‒ O. grimmiae with Grimmia pulvinata (CSN 10145). Photos
A‒I, L‒P C. Németh, J M. Vega, K L. Janošík.
298 Herzogia 36 (2), 2023

A B C

D E F

G H

I J

K L M

N O P
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete 299

Octospora gemmicola var. gemmicola agg. is characterized by narrowly ellipsoid to subfusi-

form ascospores, (17–)19 –23(–26) × 8.5 –10.5 µm, mostly with one larger and several smaller
lipid bodies. Octospora gemmicola var. gemmicola differs from O. entosthodontophila in hav-
ing somewhat shorter ascospores, a different arrangement of lipid bodies, and a different host
preference of Bryum spp. (Fig. 7F, 8F).
Octospora axillaris var. axillaris is characterized by narrowly ellipsoid ascospores that are
sometimes slightly fusiform at one or both ends, (19 –)21–26(–28) × (9 –)10 –11(–11.5) µm,
with two large lipid bodies with some accompanying smaller ones. Octospora axillaris var. ax-
illaris differs from O. entosthodontophila in having apothecia without a differentiated margin,
somewhat shorter ascospores, and Tortula acaulon as the host moss (Fig. 7G, 8G).
Octospora musci-muralis var. muralis is characterized by ellipsoid to subcylindrical ascospores
with almost parallel sided walls in the middle and broadly rounded ends, (20 –)21–28(–30) ×
(9 –)10 –11(–12) µm, mostly with two (more rarely one) large lipid bodies. O. musci-muralis
var. muralis differs from O. entosthodontophila in having ascospores with a different shape
and biseriate arrangement, as well as a different host, Grimmia pulvinata (Hedw.) Sm., grow-
ing on rocks (Fig. 7H, 8H).
Octospora musci-muralis var. neglecta is characterized by ellipsoid to subcylindrical ascospores
with broadly rounded ends, 19 –25 × 9 –11(–12) µm, mostly with two (rather rarely one) large
unequal lipid bodies. Octospora musci-muralis var. neglecta differs from O. entosthodontophila
in having ascospores with a different shape and more rounded ends, as well as growing on a dif-
ferent host, the saxicolous species Schistidium crassipilum H.H.Blom (Fig. 7I, 8I).
Octospora nemoralis Benkert & Brouwer is characterized by ellipsoid to ovoid ascospores,
(17)18 –21 × 8.5 –10.5 µm, with two unequal lipid bodies of 6 – 8 and 2– 6 µm in diameter.
Octospora nemoralis differs from O. entosthodontophila in having smaller spores and a host
preference of Fissidens bryoides Hedw. or Fissidens viridulus (Sw.) Wahlenb. (Fig. 7J, 8J).
Octospora guestfaliensis Benkert is characterized by ellipsoid to slightly fusiform ascospores,
20 –22.5 × 11.5 –13 µm, with two large lipid bodies 6 –10 µm in diameter. Octospora guest-
faliensis differs from O. entosthodontophila in having smaller ascospores and a different
host: Tortella tortuosa (Hedw.) Limpr. (Fig. 7K, 8K).
Octospora leucoloma var. leucoloma has ellipsoid ascospores, (18 –)20 –24(–26) ×
(9 –)10 –12(–13) µm, mostly with one (or two) large and several smaller lipid bodies inside.
Octospora leucoloma var. leucoloma differs from O. entosthodontophila in having a different
ascospore shape, a varying number of large lipid bodies (mostly one, rarely two vs. perma-
nently two), and a different host: Bryum argenteum Hedw. (Fig. 7L, 8L).
Octospora leucoloma var. tetraspora is characterized by mostly 4-spored (more rarely 2-, 3-,
5-, 6-, and 7-spored) asci, ellipsoid ascospores, (22–)23 –27(–30) × (10 –)11–13(–14) µm, with
one (or two) large and several small lipid bodies. Octospora leucoloma var. tetraspora differs

Fig. 8. Spores of similar species of bryophilous Pezizales with smooth ascospores and more than one lipid body. A
‒ Octospora entosthodontophila (CSN 10248), B ‒ O. gemmicola var. tetraspora (CSN 9599), C ‒ O. axillaris var.
tetraspora (CSN 9797), D ‒ O. coccinea var. coccinea (CSN 10773), E ‒ O. coccinea var. tetraspora (CSN 10269),
F ‒ O. gemmicola var. gemmicola (CSN 9815), G ‒ O. axillaris var. axillaris (CSN 10340), H ‒ O. musci-muralis var.
muralis (CSN 10225), I ‒ O. musci-muralis var. neglecta (CSN 10803), J ‒ O. nemoralis (MV20140228–05), K ‒ O.
guestfaliensis (LJ 21014), L ‒ O. leucoloma var. leucoloma (CSN 9051), M ‒ O. leucoloma var. tetraspora (CSN
10180), N ‒ O. humosa (CSN 10282), O ‒ O. itzerottii (CSN 9814), P ‒ O. grimmiae (CSN 10145), Scale bar = 20 µm.
Photos A‒I, L‒P C. Németh, J J. Eckstein, K L. Janošík.
300 Herzogia 36 (2), 2023

from O. entosthodontophila in having a lower number of ascospores in its asci, a different

ascospore shape, and Bryum argenteum as the host (Fig. 7M, 8M).
Octospora humosa s. str. has ellipsoid ascospores with rounded ends, (18 –)20 –23(–26) × 11–
13(–14) µm, with one (or two) large and several small lipid bodies. Octospora humosa differs
from O. entosthodontophila in having a different ascospore shape, a different arrangement of
lipid bodies, and Polytrichum piliferum Hedw. as the host (Fig. 7N, 8N).
Octospora itzerottii is characterized by mostly 4-spored but also 2-, 3-, 5-, and 6-spored asci,
ellipsoid ascospores, (20 –)22–27(–28) × (11–)11.5 –13(–14) µm, with mostly one (or more
rarely two) large and several small lipid bodies. Octospora itzerottii differs from O. entostho-
dontophila in having fewer and wider ascospores in its asci, as well as different hosts, i.e.
Pterygoneurum ovatum (Hedw.) Dixon and P. subsessile (Brid.) Jur. (Fig. 7O, 8O).
Octospora grimmiae Dennis & Itzerott is characterized by ellipsoid ascospores, (18 –)20 –24(–
25) × 12–14(–15) µm, with mostly one (or more rarely two) large lipid bodies. It differs from
O. entosthodontophila in having shorter and wider ascospores with mostly one large lipid body
inside and a different host, Grimmia pulvinata, a predominantly saxicolous moss (Fig. 7P, 8P).

Comparison with species known to parasitise species of the moss family

Funariaceae
Funaria hygrometrica Hedw., a ubiquitous cosmopolitan species of ruderal habitats has been
reported as the host of Lamprospora carbonicola Boud. (Benkert 1987, Döbbeler 1979),
Octospora excipulata (Benkert 1995, Döbbeler 1993, Eckstein & Eckstein 2009), and O. he-
tieri (Boud.) Dennis & Itzerott (Benkert 1995). Additionally for O. excipulata, Physcomitrium
patens (Hedw.) Mitt. and Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid. were ob-
served as hosts (Benkert 2007, Eckstein & Eckstein 2013), both growing in wet habitats
mainly along rivers and at the margins of reservoirs or pools. However, these specimens may
represent independent taxa and require further taxonomic study. Apothecia of L. carbonicola and
O. excipulata are characterized by a conspicuous membranaceous margin similar to that of O.
entosthodontophila. They clearly differ from O. entosthodontophila in microscopic features: the
ascospores of L. carbonicola are subglobose having reticulate ornamentation, and O. excipulata
has thick-walled ellipsoid ascospores with one large lipid body (Benkert 1995, Döbbeler 1993,
Eckstein & Eckstein 2009). Octospora hetieri has stiff, hyaline hairs on the margin of its apo-
thecia and ellipsoid smooth ascospores, with one large lipid body.

Acknowledgements
We are grateful to László Szabó (Hungarian Academy of Science, Institute of Materials and Environmental Chemistry)
for taking the SEM photos. We thank Domingo Chavez, Péter Finy, Attila Koszka, Miguel Ángel Ribes & Salvador Tello
for participating in some collecting trips. We would also like to thank George Greiff for his pre-submission review and
comments on the manuscript. We sincerely appreciate and thank the reviewers for their suggestions on the earlier version
of the manuscript. This study was supported by the Charles University Grant Agency project GAUK 290422.

References
Benkert, D. 1987. Beiträge zur Taxonomie der Gattung Lamprospora (Pezizales). – Zeitschrift für Mykologie 53:
195 –271.
Benkert, D. 1995. Becherlinge als Moosparasiten. – Boletus 19: 97–127.
Benkert, D. 1998. Beiträge zur Kenntnis bryophiler Pezizales-Arten. 8. Viersporige Taxa der Gattung Octospora. –
Österreichische Zeitschrift für Pilzkunde [New series] 7: 39 – 63.
Table 2. Comparison of macroscopic and microscopic features of Octospora entosthodontophila and similar taxa.
Taxon Apothecium Margin Number of Ascospore Ascospores size Lipid bodies Host (moss family) Figure
(diameter) ascospores shape
in asci, spore
arrangement
O. entosthodon- (0.5)1‒4 mm, conspicuous 8, uni- to narrowly (elon- (24.0)25.0 –25.6 –26.3(27.6) × 2 large, slightly Entosthodon spp. 7A, 8A
tophila yellow-orange membrana- subbiseriate gated) ellipsoid (10.0)10.4 –10.7–11.3(11.6) unequal ac- (Funariaceae)
or bright-orange ceous, slight- to subfusiform companied by
ly fimbriate several smaller
O. gemmicola var. (0.5 –)1–2 mm, indistinct 8, uni- to ellipsoid to nar- (21–)22–26(–30) × 2–3 large Bryum spp. (Bryaceae) 7B, 8B
tetraspora orange membeana- biseriate rowly ellipsoid (9 –)9.5 –10.5(–13) accompanied by
ceous several small
O. axillaris var. 0.8‒1.5 mm, somewhat 4, uni- to narrowly (22–)26 –32(– 40) × mostly 3, 1 large Tortula acaulon 7C, 8C
tetraspora orange pubescent biseriate ellipsoid to (9 –)10 –12(–15) encompassed by (Pottiaceae)
subfusiform 2 smaller and
accompanied by
several small
O. coccinea var. 0.5‒2 mm, somewhat 8, biseriate narrowly ellip- (20 –)24 –30(–33) × 2 large in the Bryum spp. (Bryaceae), 7D, 8D
coccinea agg. orange pubescent soid to narrowly (7–)8 –10(–11) center and 2 Encalypta vulgaris
fusiform smaller near the (Encalyptaceae),
ends Acaulon muticum
(Pottiaceae)
O. coccinea var. 1.5‒2.5 mm, somewhat 4, uni- to narrowly ellip- (24 –)30 –34(– 40) × 2 large in the Bryum spp. (Bryaceae) 7E, 8E
tetraspora orange pubescent biseriate soid to fusiform (8 –)9 –12(–15) center and 2
smaller near the
ends
O. gemmicola var. (0.5‒)1‒2 mm, indistinct 8, mostly narrowly (17–)19 –23(–26) × 8.5 –10.5 mostly 1 large Bryum spp. (Bryaceae) 7F, 8F
gemmicola agg. orange membrana- biseriate ellipsoid to and several
ceous subfusiform smaller
Németh et al.: Octospora entosthodontophila, a new smooth-spored bryophilus ascomycete

O. axillaris var. to 3 mm, orange absent 8, uni- to narrowly (19 –)21–26(–28) × 2 large and Tortula acaulon 7G, 8G
axillaris biseriate ellipsoid to (9 –)10 –11(–11.5) some smaller (Pottiaceae)
subfusiform
O. musci-muralis 1‒5 mm, orange paler 8, biseriate ellipsoid to (20 –)21–28(–30) × mostly 2 (more Grimmia pulvinata 7H, 8H
var. muralis subcylindrical (9 –)10 –11(–12) rarely 1) large (Grimmiaceae)
301
Taxon Apothecium Margin Number of Ascospore Ascospores size Lipid bodies Host (moss family) Figure
302

(diameter) ascospores shape

in asci, spore
arrangement
O. musci-mura- to 5 mm, conspicous 8, biseriate ellipsoid to 19 –25 × 9 –11(–12) mostly 2 (rather Schistidium crassipilum 7I, 8I
lis var. neglecta orange-red membrana- subcylindrical rarely 1) large (Grimmiaceae)
ceous unequal
O. nemoralis 0.2‒1.5 mm, slightly 8, uni- or ellipsoid to (17–)18 –21 × 8.5 –10.5 two large, un- Fissidens bryoi- 7J, 8J
pale-orange to fimbriate rarely bise- ovoid equal (rarely 1) des, F. viridulus
orange riate (Fissidentaceae)
O. guestfaliensis 0.5‒0.8 mm, absent 8, uniseriate ellipsoid to 20 –22.5 × 11.5 –13 2 large Tortella tortuosa 7K, 8K
orange slightly fusiform (Pottiaceae)
O. leucoloma var. 1‒3 mm, orange indistinct 8, biseriate ellipsoid (18 –)20 –24(–26) × mostly 1 (or 2) Bryum argenteum 7L, 8L
leucoloma membra- (9 –)10 –12(–13) large and several (Bryaceae)
naceous or smaller
somewhat
pubescent
and slightly
elevated
O. leucoloma var. 1‒2 mm, orange indistinct mostly 4, but ellipsoid (22–)23 –27(–30) × 1 (or 2) large Bryum argenteum 7M,
tetraspora membra- also 2, 3, 5, 6, (10 –)11–13(–14) and several (Bryaceae) 8M
naceous or and 7, uni- to small
somewhat biseriate
pubescent
and slightly
elevated
O. humosa s.str. 2‒15 mm, somewhat 8, uniseriate ellipsoid (18 –)20 –23(–26) × 1 (or 2) large Polytrichum piliferum 7N, 8N
orange pubescent 11–13(–14) and several (Polytrichaceae)
small
O. itzerottii 1‒2.5 mm, absent mostly 4, ellipsoid (20 –)22–27(–28) × mostly 1 (more Pterygoneurum spp. 7O, 8O
orange but also 2, (11–)11.5 –13(–14) rarely 2) large (Pottiaceae)
3, 5, and 6, and several
uniseriate small
O. grimmiae 1‒2 mm, orange conspicous 8, uniseriate ellipsoid (18 –)20 –24(–25) × mostly 1 (more Grimmia pulvinata 7P, 8P
to red membrana- 12–14(–15) rarely 2) large (Grimmiaceae)
ceous
Herzogia 36 (2), 2023
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Manuscript accepted: 8 June 2023.

Communicated by: Christian Berg

Addresses of the authors

Csaba Németh, Centre for Ecological Research, Institute of Ecology and Botany, Alkotmány
u. 2–4., 2163 Vácrátót, Hungary. E-mail: nemetcsaba@gmail.com
Marcel Vega, Kohlhöfen 17, DE-20355 Hamburg, Germany
Jorge Hernanz, Mastelero 12, 28033 Madrid, Spain
Jan Eckstein, Arnoldiweg 20, 37083 Göttingen, Germany. E-mail: jan.eckstein@octospora.de
Lukáš Janošík, Department of Botany, Faculty of Science, Charles University, Benátská 2, 128
01 Prague, Czech Republic. E-mail: lukas.janosik@natur.cuni.cz
Supplementary material online:
Supplementary Fig. 1. Fifty percent majority rule Bayesian phylogram obtained from the sequences of individual
loci (LSU, SSU, ITS, and EF1-α) showing the phylogenetic relationship of newly described Octospora entosthodon-
tophila with other bryophilous Pezizales. Numbers above branches represent Bayesian posterior probability scores
and RAxML bootstrap support values, respectively. GenBank accession numbers and additional collection informa-
tion are indicated in Table 1.
www.bioone.org/journals/herzogia/volume-36/issue-2/