Systematic revision of species of Atractilina and Spiropes hyperparasitic on Meliolales (Ascomycota) in the tropics

Abstract

Atractilina Dearn. & Barthol. and Spiropes Cif. are genera of asexual fungi that comprise species mainly hyperparasitic on black mildews (Meliolales, Ascomycota). Although a common group of anamorphic fungi, they have been described up to now only by morphology and their systematic position is unknown. The present study provides a morphological treatise of all known species of Atractilina and Spiropes hyperparasitic on Meliolales, including insights into their systematic position, based on DNA sequences generated here for the first time. The study was conducted, based on 33 herbarium specimens and 23 specimens recently collected in Benin and Panama. The obtained DNA sequence data (28S rDNA and ITS rDNA) of A. parasitica and of two species of Spiropes show systematic placements in the Dothideomycetes and Leotiomycetes, respectively. The sequence data of the two Spiropes spp. do not group together. Moreover, the anamorph-teleomorph connection between Atractilina parasitica and Malacaria meliolicola, a pseudothecioid fungus, is confirmed. Three species in the genus Spiropes are proposed as new to science, namely S. angylocalycis, S. carpolobiae and S. croissantiformis. Four species are reported for Benin for the first time, three species for Panama and one species for mainland America. Atractilina and Spiropes are currently two genera with highly heterogeneous species and they might have to be split in the future, once the taxonomic concepts are validated by morphology and molecular sequence data.

Full text

167
Systematic revision of species of Atractilina and Spiropes
hyperparasitic on Meliolales (Ascomycota) in the tropics
Miguel A. Bermúdez-Cova1,2 , Tina A. Hofmann3, Nourou S. Yorou4, Meike Piepenbring1
1 Mycology Research Group, Faculty of Biological Sciences, Goethe University Frankfurt Am Main, Frankfurt Am Main, Germany
2 Departamento de Biología de Organismos, División de Ciencias Biológicas, Universidad Simón Bolívar, Caracas, Venezuela
3 Centro de Investigaciones Micológicas (CIMi), Herbario UCH, Universidad Autónoma de Chiriquí, David, Panama
4 Research Unit Tropical Mycology and Plants-Soil Fungi Interactions (MyTIPS), Faculty of Agronomy, University of Parakou, BP 123, Parakou, Benin
Corresponding author: Miguel A. Bermúdez-Cova (miguelangelbermudez11@hotmail.com)
Copyright: © Miguel A. Bermúdez-Cova et al.
This is an open access article distributed under
terms of the Creative Commons Attribution
License (Attribution 4.0 International –
CC BY 4.0).
Research Article
Abstract
Atractilina Dearn. & Barthol. and Spiropes Cif. are genera of asexual fungi that com-
prise species mainly hyperparasitic on black mildews (Meliolales, Ascomycota).
Although a common group of anamorphic fungi, they have been described up to now
only by morphology and their systematic position is unknown. The present study pro-
vides a morphological treatise of all known species of Atractilina and Spiropes hyper-
parasitic on Meliolales, including insights into their systematic position, based on DNA
sequences generated here for the rst time. The study was conducted, based on 33
herbarium specimens and 23 specimens recently collected in Benin and Panama. The
obtained DNA sequence data (28S rDNA and ITS rDNA) of A. parasitica and of two
species of Spiropes show systematic placements in the Dothideomycetes and Leo-
tiomycetes, respectively. The sequence data of the two Spiropes spp. do not group to-
gether. Moreover, the anamorph-teleomorph connection between Atractilina parasitica
and Malacaria meliolicola, a pseudothecioid fungus, is conrmed. Three species in the
genus Spiropes are proposed as new to science, namely S. angylocalycis, S. carpolobi-
ae and S. croissantiformis. Four species are reported for Benin for the rst time, three
species for Panama and one species for mainland America. Atractilina and Spiropes
are currently two genera with highly heterogeneous species and they might have to
be split in the future, once the taxonomic concepts are validated by morphology and
molecular sequence data.
Key words: Anamorph-teleomorph connection, Benin, Dothideomycetes, Hyperparasit-
ism, Leotiomycetes, Panama
Introduction
Meliolales (Sordariomycetes, Ascomycota) form a large order of biotrophic,
obligate plant parasitic fungi in the Tropics and subtropics (Piepenbring et al.
2011; Hongsanan et al. 2015; Zeng et al. 2017). The order comprises two fam-
ilies, Armatellaceae and Meliolaceae, with Armatella Theiss. & Syd. and Meliola
Fr. being the most species-rich genera of each family, respectively (Hosagoudar
2003; Jayawardena et al. 2020). They are commonly known as “black mildews”,
Academic editor: Gerhard Rambold
Received:
27 November 2023
Accepted:
8 March 2024
Published:
11 April 2024
Citation: Bermúdez-Cova MA,
Hofmann TA, Yorou NS, Piepenbring
M (2024) Systematic revision of
species of Atractilina and Spiropes
hyperparasitic on Meliolales
(Ascomycota) in the tropics.
MycoKeys 103: 167–213. https://doi.
org/10.3897/mycokeys.103.115799
MycoKeys 103: 167–213 (2024)
DOI: 10.3897/mycokeys.103.115799

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
because they produce black colonies that are composed of dark, thick-walled,
branched, supercial hyphae (Rodríguez Justavino et al. 2015).
Approximately 200 species of hyperparasitic fungi, i.e. fungi parasitic on other
parasites, have been reported to grow on colonies of Meliolales (Bermúdez-Cova
et al. 2022, 2023a). These hyperparasites mainly belong to the Dothideomycetes
and the Sordariomycetes, although the systematic positions of a large number
of these fungi still remain unknown (Bermúdez-Cova et al. 2022; Bermúdez-Cova
et al. 2023a). Hyperparasitic fungi frequently overgrow entire colonies of black
mildews, so the meliolalean host may be detected only by careful search with a
light microscope (Stevens 1918; Ciferri 1955; Bermúdez-Cova et al. 2023a).
Amongst the hyperparasitic fungi, species of the anamorphic genera Atrac-
tilina Dearn. & Barthol. and Spiropes Cif. are common hyperparasites of black
mildews in the tropics. In the past, they were regarded as conidial stages of Me-
liolales (Ciferri 1955; Bermúdez-Cova et al. 2023b) and nowadays as incertae
sedis in the Ascomycota (Bermúdez-Cova et al. 2022). The genus Atractilina
includes six species of mostly hyperparasitic hyphomycetes with true synne-
mata, denticulate conidiogenous loci and pale pluriseptate conidia (Deighton
and Pirozynski 1972; Mel’nik and Braun 2013). On the other hand, the genus
Spiropes comprises 34 species of dematiaceous, mostly hyperparasitic hy-
phomycetes with mononematous, fasciculate or synnematous conidiophores
(Ellis 1968, 1971, 1976; Seifert and Hughes 2000; Bánki et al. 2023). Species
of Spiropes are characterised by the presence of conidiogenous cells with con-
spicuous, at and numerous scars, as well as pigmented conidia with 1–9 sep-
ta or pseudosepta (Ellis 1968).
Arthrobotryum Ces., Cercospora Fresen. ex Fuckel, Helminthosporium Link,
Pleurophragmium Costantin and Podosporium Schwein. are only a few of the
many genera to which species of Atractilina and Spiropes have been assigned
in the past, although they were not congeneric with the type specimens of
those genera (Ellis 1968; Deighton and Pirozynski 1972; Alcorn 1988). This re-
sulted in taxonomic uncertainty with species being transferred from one genus
to another. This problem was initially addressed by Ellis (1968) and Deighton
and Pirozynski (1972), as they did an extensive morphological revision of taxa
now assigned to Atractilina or Spiropes. For example, all the synnematous fun-
gi, hyperparasitic on Meliolales formerly assigned to the genus Arthrobotryum,
were transferred to the genus Spiropes by Ellis (1968), with the exception of
A. parasiticum (Winter) Hansf., which was transferred to the genus Atractilina
by Deighton and Pirozynski (1972).
There is currently one valid species of Atractilina, namely A. parasitica (G.
Winter) Deighton & Piroz. and 19 species of the genus Spiropes known to be
hyperparasitic on colonies of Meliolales (Ellis 1968; Deighton and Pirozynski
1972; Mel’nik and Braun 2013; Bermúdez-Cova et al. 2022). However, species
delimitation within these two genera has up to now been done by morphology
only, as species were described in the past before the molecular era and be-
cause of the challenges of isolating DNA from mixed infections (Bermúdez-Co-
va et al. 2022, 2023a, 2023b). As a result, the systematic position of both gen-
era within the Ascomycota remained unknown. The present study revises the
morphology of the species of Atractilina and Spiropes and provides the rst
insights into their systematic position according to molecular sequence data,
with emphasis on the species hyperparasitic on Meliolales.

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Materials and methods
Sample collection and morphological characterisation
Samples of leaves infected with black mildews were opportunistically collect-
ed in western Panama from January-March 2020 and in Benin in February as
well as September-October 2022. For the present study, colonies of Meliolales
hyperparasitised by Atractilina parasitica and species of Spiropes were consid-
ered. Infected leaves were dried in a plant press and deposited in the Herbarium
at the Universidad Autónoma de Chiriquí (UCH, specimens from Panama) or in
the Mycological Herbarium of the University of Parakou (UNIPAR) in Benin. Du-
plicates of large-sized samples were deposited in the Botanische Staatssam-
mlung München (M). In some cases, fungal tissue was collected prior to drying
of the specimens and preserved in CTAB buffer for subsequent DNA extraction.
Dried specimens were observed by stereomicroscopy and by light microsco-
py (LM). Measurements of at least 20 conidia and other structures have been
made for each specimen at magnications of 600× and 1000×. Measurements
are presented as mean value ± standard deviation with extreme values in paren-
theses. Line drawings were made freehand on scaled paper. Scars on conidio-
phores are drawn in surface view although further cells of the conidiophore are
drawn in optical sections. Images and drawings were edited with Photoshop
(Adobe, San Jose, California). Specimens were also analysed morphologically
by scanning electron microscopy (SEM). Materials used for SEM were prepared
according to Hofmann et al. (2010).
Host plant identication
Host plants were identied by morphological characteristics and, in some
cases, by molecular sequence data. Morphological identications were made
by comparison with herbarium specimens, literature (e.g. Akoègninou et al.
(2006); Condit et al. (2011)) and with the help of local botanists. Molecular
sequence data for species identications were obtained by polymerase chain
reaction (PCR) for the amplication of the partial region of chloroplast rbcL
with the primer pairs rbcLa-F (Levin et al. 2003) and rbcLa-R (Kress et al. 2009).
DNA was extracted from approx. 0.05 g of leaf tissue dried with silica gel using
the innuPREP Plant DNA Kit (Analytik Jena, Germany) and following the man-
ufacturer’s instructions. Protocols for PCR were carried out as described by
Fazekas et al. (2012).
DNA extraction, PCR amplication and sequencing of fungal DNA
DNA was isolated from the synnemata and hyphae of specimens using the
E.Z.N.A Forensic DNA Extraction Kit, following the manufacturer’s instructions.
To extract total genomic DNA, a small amount of clean synnemata or single co-
nidiophores were transferred into a sterile Eppendorf tube with approx. 200μl
of distilled water using sterilised tweezers and trying to avoid picking cells of
any other organism associated with the leaves and the colonies of black mil-
dews. For example, for the synnemata of Atractilina parasitica and Spiropes
melanoplaca, only the upper parts were used for DNA extraction, in order to
avoid the basal parts that are in direct contact with cells of other organisms.

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
The samples were frozen for 24 h at -20 °C, and later homogenised for 10–12
min. using a Retsch Mixer Mill MM301 with TL buffer and 2.5 mm Zirconia
beads. Isolated DNA was re-suspended in elution buffer and stored at -20 °C.
Two partial nuclear gene regions (ribosomal loci) were amplied and se-
quenced: For the large subunit nuclear ribosomal DNA (nrLSU, 28S rDNA), the
primers LR0R (Wagner and Ryvarden 2002) and LR5 (Vilgalys and Hester 1990)
were used. For the internal transcribed spacer region of ribosomal DNA (ITS),
the primers ITS5 and ITS4 (White et al. 1990) were used. The PCR mixtures con-
sisted of 1 μl genomic DNA, 15× MgCl2 reaction buffer (Bioline, Luckenwalde,
Germany), 25 mM MgCl2, 25 μM of each dNTP, 10 μM of each primer and 5 U
Taq DNA polymerase (VWR) in a total volume of 30 μl. Cycling parameters of
the PCR were as follows: initial denaturation at 94 °C for 3 min, followed by 35
cycles of amplication [denaturation at 94 °C for 30 s, primer annealing at 52°C
for 30 s and primer extension at 72 °C for 45 s] and a nal extension at 72 °C
for 5 min, followed by storage at 8 °C. PCR-products were checked on 1.5% aga-
rose electrophoresis gels containing HDGreenPlus DNA stain. Amplied PCR
products were puried with the Cycle Pure Kit (VWR-Omega, USA). Sequencing
was performed at Seqlab GmbH, Germany.
Phylogenetic analyses
Consensus sequences of trace les were generated with Geneious 10.2.2
(https://www.geneious.com, Kearse et al. 2012) and searched against Gen-
Bank (https://www.ncbi.nlm.nih.gov/, Benson et al. 2014) with MegaBLAST.
Ambiguous and miscalled bases were corrected, when possible, after exam-
ination of the corresponding chromatogram les. Sequences with a high sim-
ilarity were aligned with MAFFT v. 7 using the L-INS-i algorithm (Nakamura et
al. 2018). The alignments were manually checked by using MEGA v. 7 (Kumar
et al. 2016). Gblocks v. 0.91b (Talavera and Castresana 2007) was used to re-
move poorly-aligned positions and divergent regions from the DNA alignment.
Phylogenetic analyses of this study were conducted by applying Maximum
Likelihood (ML) in RAxML-HPC2 v.8.2.12 (Stamatakis 2014) on XSEDE (Miller
et al. 2010) and Bayesian phylogenetic inference with the programme MrBayes
3.2.6. (Ronquist et al. 2012) on XSEDE (Miller et al. 2010), available on the CIP-
RES Science Gateway web portal (http://www.phylo.org/sub_sections/portal/).
The alignment and tree are included in Suppl. material 1.
We also used T-BAS 2.1 (Carbone et al. 2019) and the “Place Unknowns” tool
to place newly-generated ITS sequences on to the Pezizomycotina tree version
2. Two FASTA les of the newly-generated ITS sequences of Spiropes were up-
loaded to the T-BAS interface. We selected the “de novo” option for the RAxML
placement, with 500 bootstrap replicates.
Results
Taxonomy
Based on morphological evidence, the hyperparasitic fungi collected in Pan-
ama and Benin are assigned to the genera Atractilina or Spiropes. Amongst
these, three species are proposed as new to science, all in the genus Spiropes.

171
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Four species represent new reports for Benin and three for Panama. We also
present a revision from herbarium material of 17 of the 19 known species of
the genus Spiropes and one species of Atractilina hyperparasitic on Meliolales.
All species synonyms, unless specied, are taken from Deighton and Pirozynski
(1972) for Atractilina parasitica and from Ellis (1968) for species of Spiropes.
Atractilina Dearn. & Barthol., Mycologia 16: 175, 1924.
Atractilina parasitica (G. Winter) Deighton & Piroz., Mycol. Pap. 128: 34, 1972
Fig. 1
≡ Arthrosporium parasiticum G. Winter, Hedwigia 25: 103, 1886.
≡ Arthrobotryum parasiticum (G. Winter) Hansf., Proc. Linn. Soc. Lond. 155: 64,
1943.
= Isariopsis penicillata Ellis & Everh., Bull. Torrey bot. Club 22: 438, 1895.
≡ Phaeoisariopsis penicillata (Ellis & Everh.) S.C. Jong & E.F. Morris, Mycopath.
Mycol. appl. 34: 271, 1968.
= Arthrobotryum tecomae Henn., Hedwigia 43: 397, 1904.
= Arthrobotryum caudatum Syd. & P. Sydow, Etudes sur la Flore du Bas et Moyen
Congo 3(1): 22, 1909.
= Arthrobotryum dieffenbachiae F. Stevens, Bot. Gaz. 65: 237, 1918.
= Atractilina callicarpae Dearn. & Barthol., Mycologia 16: 175, 1924.
= Podosporium pallidum Pat., Scient. Surv. P. Rico 8(1) Bot.: 103, 1926.
= Eriomycopsis bosquieae Hansf., Bothalia 4(2): 466, 1942.
= Arthrobotryum deightonii Hansf., Mycol. Pap. 15: 218, 1946.
= Malacaria meliolicola Syd., Annls. Mycol. 28(1/2): 69, 1930. New synonym
proposed in this study.
= Paranectria agellata Hansf., Proc. Linn. Soc. London 153(1): 28, 1941. New
synonym proposed in this study.
≡ Malacaria agellata (Hansf.) Hansf., Mycol. Pap. 15: 128, 1946. New synonym
proposed in this study.
Description. Colonies effuse, rust brown or pale brown, with hyphae that form
large, erect, dark synnemata clearly visible under the stereomicroscope, but
sometimes only loose unstalked tufts around the tips of the setae of the meli-
olalean host. Hyphae supercial, branched, septate, thin-walled, 1–2.5 µm wide,
smooth. Conidiophores may form straw-coloured or pale olivaceous synnema-
ta up to 1.5 mm long, 40 µm wide at the basal stalk-like part. Sometimes the
synnemata grow around and up the setae of the meliolalean host. Individual
conidiophores straight or sometimes exuous, cylindrical, 2.5–5 µm thick to-
wards the apex, pale olivaceous brown, with denticles. Conidia solitary, straight
or slightly curved, fusiform, truncate at the base, tapering towards the apex and
often terminating in a little bulbous swelling, 1 to mostly 3 septate, thin-walled,
variable in size, (17–)30–37(–80) × (3.5–)7–8.5 µm, at rst more or less co-
lourless, at maturity becoming pale straw coloured, minutely rough-walled. As
seen by SEM, the ornamentation of the surface of the conidia is distinctly retic-
ulated, with thin networks and no ridges.
Specimens examined. On Meliola sp. on living leaves of Opilia celtidifolia (Opil-
iaceae), Benin, Campus University of Abomey-Calavi, botanical garden, 6°25'7"N,

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Figure 1. Atractilina parasitica (MB127, MB136) a synnemata (gold spots) on colonies of Meliola sp. (black spots) on
a leaf of Opilia celtidifolia b synnemata of (gold spots) on colonies of Meliola clerodendricola (black spots) on a leaf of
Clerodendrum capitatum c synnemata d conidiophores drawn in optical section. The thickness of the wall is indicated
only in the drawing in the middle e conidia shown in optical section f–i as seen by SEM f conidiophores with denticles
ga denticle at the tip of a conidiophore h conidium i bulbous swelling at the tip of a conidium. Scale bars: 1.5 mm (b);
1mm (c); 5 μm (d,e,i); 8 μm (f); 1 μm (g); 6 μm (h).
2°20'34"E, 24 m a.s.l., 9 February 2022, M. A. Bermúdez-Cova, A. Tabé, D. Dong-
nima, O.P. Agbani, M. Piepenbring, N.S. Yorou, MB127 (UNIPAR, M); on Meliola
clerodendricola on living leaves of Clerodendrum capitatum (Lamiaceae), Benin,
Abomey-Calavi, Zopah, 6°30'8"N, 2°20'24"E, 37 m a.s.l., 12 February 2022, M. A.
Bermúdez-Cova, A. Tabé, D. Dongnima, O.P. Agbani, M. Piepenbring, N.S. Yorou,
MB133; on Meliola clerodendricola on living leaves of Clerodendrum capitatum,
Benin, Allada, Sékou, 6°38'56"N, 2°11'38"E, 48 m a.s.l., 12 February 2022, M. A.
Bermúdez-Cova, A. Tabé, D. Dongnima, O.P. Agbani, M. Piepenbring, N.S. Yorou,
MB136 (UNIPAR, M, GenBank accession number: OR804686); on Meliola sp. on
living leaves of Pterocarpus santalinoides (Fabaceae), Benin, Lokoli, border of
forest, 7°3'41"N, 2°15'26"E, 22 m a.s.l., 20 February 2022, M. A. Bermúdez-Cova,
A. Tabé, D. Dongnima, L. Konetche, M. Piepenbring, R. Hounkarin, MB160 (M);

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
on Meliola sp. on living leaves of Coffea arabica (Rubiaceae), Benin, Attogon,
Niaouli, CRA-Sud center, 6°44'24"N, 2°8'25"E, 122 m a.s.l., 28 February 2022, M.
A. Bermúdez-Cova, A. Tabé, I. Agonglo, M. Piepenbring, N.S. Yorou, O.P. Agbani,
MB178 (UNIPAR, M, GenBank accession numbers: OR804685 and OR804687);
on Meliola sp. on living leaves of Coffea arabica, Benin, Atlantique, Attogon,
Niaouli Forest, 6°44'23"N, 2°8'26"E, 119 m a.s.l., 19 September 2022, A. Krauß,
A. Tabé, O. Koukol, N.S. Yorou, AK06H (UNIPAR, M, GenBank accession number:
OR804684); on Meliola sp. on living leaves of Clerodendrum capitatum, Benin,
Atlantique, Attogon, Pahou Forest, 6°22'56"N, 2°9'35"E, 13 m a.s.l., 6 October
2022, A. Krauß, A. Tabé, O. Koukol, N.S. Yorou, AK61.
Additional specimens examined. On Meliola lasiotricha on leaves of un-
known plant host, Puerto Rico, 1926, M.B. Ellis (IMI 130722, type specimen
of Podosporium pallidum); On Meliola clerodendri on leaves of Clerodendrum
cyrtophyllum, Taiwan, 1938, W. Yamamoto (IMI 31921b, type specimen of At-
ractilina parasitica).
Illustrations. This species was illustrated by Deighton and Pirozynski (1972).
Known hosts and distribution. On colonies of Amazonia spp., Asteridiella
spp., Irenopsis spp. and Meliola spp. on living leaves of various plants in Congo,
Ghana, Guinea, India, Mauritius, Nigeria, Perú, Philippines, Puerto Rico, Sierra
Leone, St. Thomé, Taiwan, Tanzania, Uganda, U.S.A. and Venezuela. Only one
single collection on Balladyna sp. (Balladynaceae, Dothideomycetes) as a fun-
gal host (Deighton and Pirozynski 1972). Atractilina parasitica is reported here
for the rst time for Benin.
Notes. Only two species of the genus Atractilina with hyperparasitic lifestyle
are known, namely A. asterinae and A. parasitica (Deighton and Pirozynski
1972). Atractilina asterinae differs from A. parasitica by the presence of 3–10
septate, thick-walled conidia.
The specimens of A. parasitica collected on leaves of Coffea arabica
(MB 178, AK4H, AK06H) were found growing together with pseudothecia of
Malacaria meliolicola Syd. (Tubeuales, Dothideomycetes). According to
Hansford (1941, as Paranectria agellata; 1946), M. agellata is most probably
the perfect state of A. parasitica. The specimens collected by Hansford were
also growing on coffee leaves. The latter and the fact that the DNA sequenc-
es we obtained from A. parasitica (GenBank accession numbers: OR804684,
OR804686, OR804685 and OR804687) and M. meliolicola (GenBank accession
numbers: OR805247 and OR805248) clustered together in one single strong-
ly-supported clade (Fig. 22), conrm the anamorph-teleomorph connection be-
tween both species. For an updated species description of M. meliolicola, see
Bermúdez-Cova et al. (2023b).
Spiropes Cif., Sydowia 9(1–6): 302, 1955
Spiropes angylocalycis Berm.-Cova & M. Piepenbr., sp. nov.
MycoBank No: 850990
Fig. 2
Holotype. On Meliola sp. on living leaves of Angylocalyx oligophyllus (Fabaceae),
Benin, Atlantique, Attogon, Niaouli Forest, 6°44'42"N, 2°7'50"E, 69 m a.s.l.,

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Figure 2. Spiropes armatellae (MB 167) a, b conidiophores growing intermingled with hyphae of Meliola sp. on leaves of
Angylocalyx oligophyllus c conidiophore with scars d conidia shown in optical section. The thickness of the wall is shown
in the two drawings on the right-hand side e, f as seen by SEM e part of a conidiophore with scar f conidium. Scale bars:
0.3 mm (a); 0.2 mm (b); 5 μm (c, d); 2 μm (e); 7 μm (f).
28 February 2022, M.A. Bermúdez, A. Tabé, D. Dongnima, I. Agonglo, O.P. Agba-
ni, M. Piepenbring, N.S. Yorou, MB167 (M).
Etymology. Named after the genus of the host plant.
Description. Colonies effuse, dark brown to black, velvety to hairy. Hyphae
supercial, branched, anastomosing, septate, 0.5–2 µm wide, straw-coloured,
smooth. Conidiophores arising singly, erect or ascending, straight to exuous,
mostly exuous at the tips, septate, up to 350 µm long, 4–6 µm thick, pale oliva-
ceous-brown to brown, with rough surface, with scattered scars mostly in upper
parts of the conidiophores. Conidia solitary, straight or slightly curved, fusiform
to obclavate, 3–septate, (15–)17–25(–30) × 5–6.5 µm, 2–3 µm wide at the
base, brown, the cells at each end pale brown, septa darker in colour, verrucose.
As seen by SEM, the ornamentation of the spores is distinctly reticulated, with
thin to thick networks and no ridges.
Known distribution. On colonies of Meliola sp. on living leaves of Angyloca-
lyx oligophyllus in Benin.
Notes. Spiropes angylocalycis is similar to S. clavatus by the presence of 3–
septate mostly fusiform conidia, with a similar size range (Ellis 1968). However,
the conidiophores of S. clavatus are synnematous, while they are mononema-
tous in S. angylocalycis.

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Spiropes armatellae M.B. Ellis, Mycol. Pap. 125: 15, 1971
Fig. 3
Type. On Armatella cinnamomicola on leaves of Cinnamomum sp. (Lauraceae),
Sri Lanka, Ceylon, 1971, M.B. Ellis (IMI134405b. The type specimen was not
available for loan).
Description. Colonies effuse, dark brown to black, hairy. Hyphae supercial,
branched, septate, 1–3 µm wide, straw-coloured, smooth. Conidiophores aris-
ing singly, erect or ascending, straight to exuous, mostly exuous at their tips,
septate, up to 300 µm long, 5–8 µm thick, brown to dark brown, paler towards
the apex, with rough surface, with scattered scars in upper parts of the conid-
iophores. Conidia solitary, straight or slightly curved, obclavate to obpyriform,
mostly 1–septate, (20–)30–42(–50) × (6–)7–8(–10) µm, 2–3.5 µm wide at the
base, brown, paler towards the ends, verrucose when seen by LM and SEM.
Specimen examined. On Armatella litseae on leaves of Daphnidium pulcher-
rimum (Lauraceae), India, west Bengal, 1967, M.K. Maity (IMI 136371); on Ar-
matella cinnamomicola on leaves of Cinnamomum sp., Myanmar, Thaton, 1971,
M.M. Thaung, (IMI 161265).
Known hosts and distribution. On colonies of Armatella spp. on various
plants in India, Myanmar and Sri Lanka (Ellis 1971).
Illustrations. This species was illustrated by Ellis (1971).
Notes. Two known species of Spiropes are hyperparasitic on species of the
genus Armatella (Meliolales, Armatellaceae), namely S. armatellae and S. ar-
matellicola (Ellis 1971, Hosagoudar et al. 2002). According to Hosagoudar et
Figure 3. Spiropes armatellae (IMI 161265) a conidiophores with young conidium b, c conidia b shown in optical section.
The thickness of the wall is indicated only in the drawing on the left-hand side c as seen by SEM. Scale bars: 5 μm (a);
2.5 μm (b); 10 μm (c).

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al. (2002), both species are similar, but differ by the ornamentation of the conid-
ia. The conidia of S. armatellicola are smooth, while those of S. armatellae are
distinctly reticulated. However, it is sometimes dicult to observe the surface
of the conidia by LM. Therefore, we recommend to analyse the ornamentation
of the spores of S. armatellicola by SEM. The scars of S. armatellae could not be
observed by SEM and it is necessary to collect fresh specimens of this fungus
for further morphological analysis.
Spiropes armatellicola M.B. Ellis, Mycol. Pap. 125: 15, 1971
Type. On Armatella sp. on leaves of Actinodaphne sp. (Lauraceae), Banasur-
an Hills, Wyanad, Kerala, India, 16 April 1999, C.K. Biju (HCIO 43621. The type
specimen was not available for loan by HCIO).
Species description. This species was described by Hosagoudar et al.
(2002).
Known hosts and distribution. On colonies of Armatella sp. on living leaves
of Actinodaphne sp. in India (Hosagoudar et al. 2002).
Illustrations. This species was illustrated by Hosagoudar et al. (2002).
Notes. This species is only known from the type specimen.
Spiropes capensis (Thüm.) M.B. Ellis, Mycol. Pap. 114: 5, 1968
Fig. 4
≡ Cercospora capensis (Thüm.) Sacc., Syll. fung. 4: 469, 1886.
≡ Helminthosporium capense (Thüm.) [as ‘Helmisporium’], Flora, Regensburg
59: 570, 1876.
≡ Pleurophragmium capense (Thüm.) S. Hughes, Can. J. Bot. 36: 796, 1958.
= Helminthosporium carpocrinum Cif. [as ‘Helmisporium’], Annls. Mycol. 36(2/3):
236, 1938.
= Helminthosporium coffeae Massee [as ‘Helmisporium’], Bull. Misc. Inf., Kew:
167, 1901.
≡ Sporhelminthium coffeae (Massee) Speg., Physis, Rev. Soc. Arg. Cienc. Nat.
4(no. 17): 292, 1918.
= Helminthosporium ci H.S. Yates [as ‘cuum’], Philipp. J. Sci. (Bot.) 13: 382,
1918.
= Helminthosporium cinum Sacc. [as ‘Helmisporium’], Atti Accad. Sci. Ven.-
Trent.-Istr., Sér. 3, 10: 90, 1919.
= Helminthosporium fumagineum Sacc. [as ‘Helmisporium’], Atti Accad. Sci.
Ven.-Trent.-Istr., Sér. 3, 10: 90, 1919.
= Helminthosporium licicola Henn., Hedwigia 44: 71, 1905.
= Helminthosporium glabroides F. Stevens [as ‘Helmisporium’], Bot. Gaz. 65(3):
240, 1918.
= Helminthosporium melioloides Sacc. [as ‘Helmisporium’], Atti Accad. Sci.
Ven.-Trent.-Istr., Sér. 3, 10: 89, 1919.
= Helminthosporium orbiculare Lév., Annls. Sci. Nat., Bot., Sér. 3, 5: 299, 1846.
= Helminthosporium philippinum Sacc. [as ‘Helmisporium’], Atti Accad. Sci.
Ven.-Trent.-Istr., Sér. 3, 10: 89, 1919.

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= Helminthosporium subsimile Sacc., Boll. Orto bot., Napoli 6: 23, 1921.
= Helminthosporium tapurae Allesch., Hedwigia 36(4): 245, 1897.
= Napicladium portoricense Speg., Boln Acad. nac. Cienc. Córdoba 26(2–4):
363, 1921.
≡ Helminthosporium portoricense (Speg.) Cif., Sydowia 9(1–6): 298, 1955.
= Nascimentoa pseudoendogena Cif. & Bat., Publicações Inst. Micol. Recife
44:4, 1956.
Description. Colonies effuse, dark brown to black, hairy (Ellis 1968). Hyphae
supercial, branched, septate, 2–4 µm wide, pale olive to olivaceous-brown,
smooth. Conidiophores arising singly or in groups, sometimes in large groups
of 50–100 conidiophores, terminally or laterally from the hyphae, erect or as-
cending, straight or exuous, septate, up to 600 µm long, 5–9 µm thick along
most of their length, brown to dark brown, paler closer to the apex, with termi-
nal and lateral scars. Conidia solitary, straight or curved, fusiform to obclavate,
truncate at the base, 3–6 (usually 4 or 5) pseudosepta, (33–)50–60(–78) ×
(5.5–)6–11(–16) µm, 1–4 µm wide at the base, light brown to brown, smooth.
Specimen examined. On Meliola sp. on living leaves of Coffea arabica, Benin,
Atlantique, Attogon, Niaouli Forest, 6°44'23"N, 2°8'26"E, 119 m a.s.l., 19 Sep-
tember 2022, A. Krauß, A. Tabé, O. Koukol, N.S. Yorou, AK06H.
Additional specimens examined. – On leaves of Ficus ulmifolia (Moraceae),
Philippines, Los Baños, 1915, C.F. Baker, 451 (IMI 130940, type of Helminthospori-
um fumagineum); on Meliola compositarum on leaves of Eupatorium portoricense
Figure 4. Spiropes capensis (AK06H) a, b groups of conidiophores growing on hyphae of Meliola sp c conidiophores
growing on hyphae of Meliola sp. shown in optical section d conidia shown in optical section. The thickness of the outer
wall layer is indicated only in the drawing on the right-hand side e, f as seen by SEM e conidiophores with scars f conidia.
Scale bars: 1 mm (a, b); 8.5 μm (c); 5 μm (d); 5 μm (e); 20 μm (f).

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
(Asteraceae), Puerto Rico, Bega Vaja, 1921, no. 1753 (IMI 100331a, type of Napi-
cladium portoricense).
Known hosts and distribution. On colonies of Appendiculella spp., Asteri-
diella spp., Irenopsis spp. and Meliola spp. on living leaves of various plants in
Amboina, Bolivia, Brazil, Cameroon, Congo, Dominican Republic, Ghana, India,
Jamaica, Malaya, Peru, Philippines, Puerto Rico, Sabah, Sierra Leone, South Af-
rica, Tanzania, Trinidad, Uganda and Venezuela (Ellis 1968); on Meliola sp. on
living leaves of Coffea arabica in Benin (this study). Spiropes capensis is report-
ed here for the rst time for Benin.
Illustrations. This species was illustrated by Ellis (1968).
Notes. According to the nomenclatural and taxonomic database Index Fun-
gorum (http://www.IndexFungorum.org), the current name of the Spiropes cap-
ensis is Pleurophragmium capense (Thüm.) S. Hughes. The genus Pleurophrag-
mium (incertae sedis, Ascomycota) was established by Costantin (1888) and
it comprises species with brown to dark brown conidiophores and sympodially
proliferating, denticulate conidiogenous cells producing holoblastic, simple,
mostly 3–septate, brown to dark brown conidia (Abarca et al. 2007). According
to Ellis (1968), the at double scar is a good taxonomic character to distinguish
species of Spiropes from Pleurophragmium, since, in the latter, the conidia are
borne at the tips of tapered denticles. The morphological analysis of our sam-
ples and the type specimens (AK06H, IMI 100331a and IMI 130940) revealed
the presence of at double scars (Fig. 4e) and no denticles. We think that the
examined species differs morphologically from species in the genus Pleuro-
phragmium and, therefore, it should be retained in the genus Spiropes.
Spiropes caribensis Hol.-Jech., Česká Mykol. 38(2): 113, 1984
Fig. 5
Description. Colonies effuse, dark brown to black, velvety to hairy. Hyphae su-
percial, branched, septate, 1.5–3.5 µm wide, pale olivaceous-brown, smooth.
Conidiophores arising singly, erect or ascending, straight or exuous, sep-
tate, up to 240 µm long, 4–8 µm thick, pale brown to brown, smooth, with few
scars. Conidia solitary, straight or slightly curved, obclavate, central cells bar-
rel-shaped, 3-septate, (30–)36–48(–41.5) × (7.5–)9.5–11.5 µm, 4.5–6 µm wide
at the truncate base, the central cells pale brown, the cells at the ends paler and
almost hyaline, smooth.
Specimen examined. On Meliola sp. on leaves of an unknown palm-tree,
Cuba, Isla de La Juventud (= Isla de Pinos), Los Indios, south-west of La Caña-
da, 1981, V. Holubová-Jechová (PRM 831531, holotype).
Known hosts and distribution. On Meliola sp. on living leaves of an unidenti-
ed palm tree in Cuba (Holubová-Jechová and Sierra 1984).
Illustrations. This species was illustrated by Holubová-Jechová and Sierra
(1984).
Notes. Spiropes caribensis is similar to S. helleri, but differs from the lat-
ter by paler conidia, with wider truncate base (S. helleri has conidia with a
truncate base 3–4 µm wide) and shorter conidiophores (up to 600 µm long
in S. helleri; Holubová-Jechová and Sierra (1984)). As seen by SEM, conidia
of S. caribensis are smooth (Fig. 5b), while conidia of S. helleri are distinctly

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
reticulated (Fig. 13e). The scars could not be observed by SEM and it is, there-
fore, necessary to collect fresh specimens of this fungus for further morpho-
logical analyses. S. caribensis is only known from the type specimen.
Spiropes carpolobiae Berm.-Cova & M. Piepenbr., sp. nov.
MycoBank No: 850987
Fig. 6
Holotype. On Meliola cf. carpolobiae on living leaves of Carpolobia lutea (Poly-
galaceae), Benin, Atlantique, Attogon, Niaouli Forest, 6°44'41"N, 2°7'52"E, 68 m
a.s.l., 28 February 2022, M.A. Bermúdez, A. Tabé, D. Dongnima, I. Agonglo, O.P.
Agbani, M. Piepenbring, N.S. Yorou, MB166 (M).
Etymology. Named after the genus of the host plant.
Description. Colonies effuse, dark brown to black, velvety to hairy. Hyphae
supercial, branched, anastomosing, septate, 1–2 µm wide, straw-coloured,
smooth. Conidiophores arising singly, erect or ascending, straight to exu-
ous, septate, up to 250 µm long, 2–5 µm thick, sometimes thicker at the apex,
brown, not smooth, with scattered scars mostly in the upper parts of the conid-
iophores. Conidia solitary, straight or slightly curved, ovate to slightly fusiform,
3–septate, (12.5–)13–16(–19) × 5–7 µm, 2–2.5 µm wide at the base, brown,
the cells at each end pale brown, septa darker, surface verrucose. As seen by
SEM, the ornamentation of the conidia is distinctly reticulated, with thin to thick
networks that can form ridges.
Known distribution. On colonies of Meliola cf. carpolobiae on living leaves of
Carpolobia lutea in Benin.
Notes. S. carpolobiae is the only known species of Spiropes with ovate to
slightly fusiform conidia.
Spiropes clavatus (Ellis & Martin) M.B. Ellis, Mycol. Pap. 114: 25, 1968
Fig. 7
≡ Isariopsis clavata Ellis & Martin, Am. Nat. 18: 188, 1884.
Figure 5. Spiropes caribensis (PRM 8311531) a conidia shown in optical section b, c as seen by SEM b conidium c basis
of a conidium with a at scar. Scale bars: 10 μm (a); 9 μm (b); 4 μm (c).

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≡ Arthrobotryum clavatum (Ellis & Martin) Höhn, Sber. Akad. Wiss. Wien, Math.-
naturw. Kl., Abt. 1, 125: 120, 1916.
≡ Bitunicostilbe clavata (Ellis & Martin) M. Morelet, Bull. Soc. Sci. nat. Arch.
Toulon et du Var 7: 195, 1971.
= Podosporium chlorophaeum Speg., An. Mus. nac. Hist. nat. B. Aires 20: 450,
1910.
= Arthrobotryum noz-moscatae Bat. & J. Silva, Anais IV Congr. Soc. bot. Brasil:
144, 1953.
Description. Colonies effuse, brown to dark brown or black. Hyphae super-
cial, branched, anastomosing, septate, 1–3 µm wide, pale olivaceous-brown.
Conidiophores tightly packed to form dark brown to blackish synnemata up to
700 µm long, 20–40 µm thick, often splaying out to a width of up to 110 µm at
the apex. Individual hyphae straight or exous, cylindrical, 1–3 µm thick near
the base, 4–7 µm thick near the apex, dark brown, paler towards the apex, ver-
rucose, with numerous conidial scars. Conidia solitary, fusiform to obclavate,
mostly 3–, rarely 1–, 2– or 4–septate, (13–)18–25(–33) × (4–)5–7(–8) µm,
tapering to about 1–1.5 µm at the apex and at the base, pale brown to brown,
Figure 6. Spiropes carpolobiae (MB 166) a conidiophores growing intermingled with hyphae of Meliola sp. on a leaf of Car-
polobia lutea b conidiophore with scars c Conidia shown in optical section. The thickness of the wall is shown in the left-hand
drawing d, e as seen by SEM d conidiophore with scar e conidium. Scale bars: 0.3 mm (a); 5 μm (b, c); 5 μm (d); 3 μm (e).

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the cells at each end paler, wrinkled. As seen by SEM, the ornamentation of the
spores is distinctly reticulated, with thin to thick networks and no ridges.
Specimens examined. On Meliola panici on leaves of Panicum glutinosa,
Puerto Rico, El Alto de la Bandera, 1913, F.L. Stevens & W.E. Hess, n°4368 (IMI
130764); on Meliola sp. on leaves of Raphia monbuttorum, Uganda, 1915, R.
Dümmer, (IMI 102772); on Meliola thouiniae on leaves of an unknown plant,
Brasil, São Paulo, 1940, A.R. Campos (IMI 130975, type of Arthrobotryum
noz-moscatae).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliolales on living leaves of
various plants in Argentina, Brazil, Ghana, Malaysia, Puerto Rico, Sierra Leone,
Trinidad and Uganda (Ellis 1968).
Notes. In the nomenclatural and taxonomic database Index Fungorum
(http://www.IndexFungorum.org), the current name of the Spiropes clavatus
is Bitunicostilbe clavata (Ellis & Martin) M. Morelet. The genus Bitunicostilbe
(incertae sedis, Ascomycota) was proposed by Morelet (1971) to accom-
modate two species, namely B. clavata and B. linderae, that were previously
cited in other genera. Although the publication by Morelet was not available
for this study, the morphological analysis of the herbarium specimens (IMI
130764, 130975) revealed that the features of these specimens are consis-
tent with the description of Spiropes clavatus by Ellis (1968). The species
has typical characteristics of the genus Spiropes, such as at double scars
(Fig. 7c) and, therefore, it should be classied in this genus. De Beer et al.
(2013) analysed the type and additional specimens of B. linderae (as Graphi-
um linderae) and concluded that this species should be also classied in the
genus Spiropes.
Figure 7. Spiropes clavatus (IMI 102772) a conidiophores with scars b conidia shown in optical section c, d as seen by
SEM c conidiophore with scars d conidium. Scale bars: 5 μm (a); 2.5 μm (b); 1 μm (c); 5 μm (d).

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Spiropes croissantiformis Berm.-Cova & M. Piepenbr., sp. nov.
MycoBank No: 850984
Fig. 8
Holotype. On Meliola cf. xylopiae on living leaves of Xylopia frutescens, Pana-
ma, Chiriquí Province, Cochea, Cochea River Trail, 8°32'37"N, 82°23'03"W, 181 m
a.s.l., 26 February 2020, M.A. Bermúdez, A. Sanjur, A. Villarreal, MB110 (UCH).
Etymology. Named after the shape of the conidia.
Description. Colonies effuse, dark brown to black, with tightly packed hy-
phae that form erect, dark synnemata clearly visible under the stereomicro-
scope. Hyphae supercial, branched, septate, 1–2 µm wide, straw-coloured,
smooth. Conidiophores tightly packed to form dark brown to blackish synne-
mata up to 400 µm high, spreading out at the apex, up to 80 µm diam. Individual
hyphae mostly straight, cylindrical, 3–5 µm thick, with numerous small scars,
brown, paler towards the apex, rough. Conidia straight or curved, mostly cres-
cent-shaped, sometimes fusiform, mostly 3(–5)–septate, (14–)20–24(–33) ×
(3.5–)5–6.5 µm, with two golden brown middle cells and paler cells at each. As
Figure 8. Spiropes croissantiformis (MB 110) a synnemata (indicated by white arrows) growing on colonies of Meliola cf.
xylopiae b synnema (indicated by a black arrow) c conidiophores with scars and young conidia, shown in optical section
d conidia shown in optical section. The thickness of the wall is only shown for the rst spore from the left e, f as seen by
SEM e part of a conidiophore with scars f conidia. Scale bars: 160 μm (a); 400 μm (b); 5 μm (c, d); 5 μm (e); 10 μm (f).

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seen by SEM, the ornamentation of the spores is distinctly reticulated, with thin
to thick networks and no ridges.
Known distribution. On colonies of Meliola cf. xylopiae on living leaves of
Xylopia frutescens (Annonaceae) in Panama.
Notes. Spiropes xylopiae is a synnematous hyperparasitic species of
Spiropes with the shortest synnemata (up to 400 µm), when compared to other
synnematous species, such as S. melanoplaca with synnemata that can reach
up to 1.5 mm and S. penicillium with synnemata up to 700 µm high. In addition
to this, the new species has crescent-shaped conidia, a feature that is not pres-
ent in any other known species of the genus.
Spiropes deightonii M.B. Ellis, Mycol. Pap. 114: 18, 1968
Fig. 9
Description. Colonies effuse, olive to olivaceous-brown, velvety or hairy. Hyphae
supercial, branched, septate, 0.5–2 µm wide, pale olive to olivaceous-brown,
smooth. Conidiophores arising singly or in groups terminally or laterally from
the hyphae, erect or ascending, straight or exous, septate, up to 400 µm long,
2–4 µm thick along most of their length, swollen towards the apex, 5–8 µm
thick, brown, reticulate as seen by SEM, with scattered cylindrical scars. Conidia
solitary, straight or slightly curved, obovate to clavate, truncate at their base, 3–
septate, (10–)12–14(–15) × (5–)6–8 µm, 1.5–2 µm wide at the base, the cells
Figure 9. Spiropes deightonii (IMI48956a) a conidiophores b conidia, as seen by LM (two upper spores; the thickness of
the wall is indicated only in the drawing on the left-hand side) and by SEM (bottom spore) c, d as seen by SEM c conidio-
phore d conidia. Scale bars: 5 μm (a, b); 8 μm (c); 5 μm (d).

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at each end of a conidium subhyaline or pale brown, intermediate cells brown,
ornamented. As seen by SEM, the ornamentation of the spores is distinctly
reticulated, with thin to thick networks that can form ridges.
Specimen examined. On Meliola borneensis on Uvaria chamae, Sierra Leone,
1951, F.C. Deighton, (IMI 48956a, type of S. deightonii).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliola borneensis on living
leaves of Uvaria chamae (Annonaceae) in Sierra Leone (Ellis 1968).
Notes. Spiropes deightonii and Spiropes intricatus are the only known spe-
cies of the genus that present conidiophores that swell in the areas where co-
nidia are formed (Figs 9, 14; Ellis (1968)). Spiropes intricatus differs from S.
deightonii by the presence of larger conidia (16–23 µm long) that are more
oblong-ellipsoid (Ellis 1968), rather than obovate or clavate. S. deightonii is only
known from the type specimen.
Spiropes dorycarpus (Mont.) M.B. Ellis, Mycol. Pap. 114: 27, 1968
Fig. 10
≡ Helminthosporium dorycarpum Mont., Annls Sci. nat., 2 Sér., 17: 120, 1842.
≡ Pleurophragmium dorycarpum (Mont.) Hughes, Can. J. Bot. 36: 797, 1958.
= Helminthosporium orbiculare Lév., Annls Sci. nat., 3 Sér., 5: 299, 1846.
= Napicladium myrtacearum Speg., An. Soc. cient. Argent. 26: 71, 1888.
≡ Sporhelminthium myrtacearum (Speg.) Speg., Physis 4(17): 292, 1918.
= Helminthosporium conspicuum McAlpine, Proc. Linn. Soc. N.S.W. 22: 40, 1897.
= Podosporium densum Pat., J. Bot. Paris 11: 373, 1897.
= Helminthosporium asterinoides Sacc. & P. Syd., apud Saccardo, Rc. Congr.
Bot. Palermo, May 1902: 58, 1902.
≡ Sporhelminthium asterinoides (Sacc. & Syd.) Speg., Physis 4(17): 292, 1918.
= Helminthosporium melastomacearum F. Stevens, Bot. Gaz. 65: 242, 1918.
= Helminthosporium panici F. Stevens, Bot. Gaz. 65: 242, 1918.
= Helminthosporium parathesicola [as ‘parathesicolum’] F. Stevens, Bot. Gaz.
65: 242, 1918.
Description. Colonies effuse, brown to dark brown, hairy. Hyphae supercial,
branched, septate, 1–3 µm wide, straw-coloured, pale brown, smooth. Conidio-
phores arising singly or in groups, terminally or laterally from the hyphae, erect or
ascending, straight or exous, septate, up to 700 µm long, 3–7 µm thick, straw-co-
loured, pale brown to brown, with scattered cylindrical scars towards the apex.
Conidia solitary, straight or slightly curved, variable in shape, but mostly obclav-
ate to fusiform, truncate at the base, mostly 3–septate, but sometimes with 4 to
5 septa, (16–)20–35(–40) × (4.5–)5–7 µm, straw-coloured to pale brown, middle
cells slightly darker, wrinkled or verrucose. As seen by SEM, the ornamentation
of the spores is distinctly reticulated, with thin to thick networks and no ridges.
Specimen examined. On Meliola sp. on living leaves of Coffea arabica, Benin,
Atlantique, Attogon, Niaouli Forest, 6°44'23"N, 2°8'26"E, 119 m a.s.l., 19 Sep-
tember 2022, A. Krauß, A. Tabé, O. Koukol, N.S. Yorou, AK06H.
Additional specimens examined. On Eugenia pungens, Brasil, Guarapí, 1883,
B. Balansa, 3939, (IMI 100322, type of Napicladium myrtacearum); on Meliola sp.

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on leaves of an unknown plant, Cuba, R. de la Sagra (IMI 10002, type of Hel-
minthosporium dorycarpum).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Appendiculella spp., Asteri-
diella spp., Clypeolella spp., Irenopsis spp., Meliola spp. and Schiffnerula spp.,
on living leaves of various plants in Australia, Brazil, Chile, Congo, Cuba, Do-
minican Republic, Ghana, Guyana, India, Malaysia, Nigeria, Puerto Rico, Sierra
Leone, South Africa, Taiwan, Tanzania and Uganda (Ellis 1968). Spiropes dory-
carpus is reported here for the rst time for Benin.
Notes. Spiropes dorycarpus is similar to S. effusus and S. helleri by the presence
of non-synnematous conidiophores and conidia mostly with three true septa. How-
ever, conidia of S. effusus are narrower (3–5 µm) than those of S. helleri (7–13 µm).
Spiropes effusus (Pat.) M.B. Ellis, Mycol. Pap. 114: 10, 1968
Fig. 11
≡ Podosporium effusum Pat., Scient. Surv. P. Rico 8(1): 103, 1926.
Figure 10. Spiropes dorycarpus (AK06H) a supercial hyphae growing on a colony of Meliola sp. on a leaf of Coffea arabi-
ca b, c in optical section b conidiophore growing on a hypha of Meliola sp. c conidia. The thickness of the wall is indicated
only in the drawing on the left-hand side d, e As seen by SEM d conidiophore with a scar e conidium. Scale bars: 1 mm (a);
5 μm (b); 3.5 μm (c); 3 μm (d); 7 μm (e).

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= Helminthosporium dorycarpum var. amazoniae Hughes [as ‘Helmisporium’],
Mycol. Pap. 50: 24, 1953.
≡ Pleurophragmium dorycarpum var. amazoniae (S. Hughes) S. Hughes, Can. J.
Bot. 36: 797, 1958.
Description. Colonies effuse, olive to brown, hairy. Hyphae supercial, branched,
septate, 1–2 µm wide, yellowish, olive or pale brown, smooth. Conidiophores
arising singly or in groups, as terminal and lateral branches on the hyphae,
erect, straight or exous, septate, up to 300 µm long, 3–4 µm thick, slightly
reticulated when seen by SEM, with few or many small conidial scars towards
the apex. Conidia solitary, narrowly obclavate to fusiform, truncate at the base,
mostly 3(–5)–septate, (15–)20–36 × (3–)3.8–4.5(–5) µm, pale brown, the cen-
tral cells slightly darker, verruculose. As seen by SEM, the ornamentation of the
spores is distinctly reticulated, with thin networks and no ridges.
Specimen examined. On meliolalean fungus on leaves of Piper sp., Puerto
Rico, Río Piedras, 1926, Heller, 142 (IMI 130721, type of Podosporium effusum);
on Amazonia psychotriae on leaves of Psychotria warneckei, Ghana, Togoland,
1938, F.C. Deighton M1617B (IMI 9996a).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliolales, especially Amazo-
nia spp., on living leaves of various plants in Ghana, Puerto Rico, Sierra Leone
and Venezuela. One record on Asterina sp. (Asterinales, Ascomycota) in Ugan-
da (Ellis 1968).
Figure 11. Spiropes effusus (IMI 130721) a conidiophore shown in optical section b conidia. The rst two drawings show
spores in optical section. The right-hand drawing shows a conidium as seen by SEM c, d as seen by SEM c conidiophore
with scars and conidia d conidium. Scale bars: 5 μm (a); 8 μm (b); 2 μm (c); 8 μm (d).

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Notes. Spiropes effusus has conidia similar in size to those of S. dorycarpus.
However, conidia of S. dorycarpus are wider (5–7 µm) than in S. effusus.
Spiropes fumosus (Ellis & Martin) M.B. Ellis, Mycol. Pap. 114: 20, 1968.
≡ Helminthosporium fumosum Ellis & Martin, Am. Nat. 18: 70, 1884.
≡ Brachysporium fumosum (Ellis & Martin) Sacc., Syll. Fung. 4: 428, 1886.
Type. On Meliola sp. on leaves of Persea palustris (Lauraceae), Florida, U.S.A,
1883, G. Martin (NY 830274. The type specimen was not available for loan
by NY).
Species description. This species was described by Ellis (1968).
Known hosts and distribution. On colonies of Meliola sp. on living leaves of
Persea palustris in the U.S.A. (Ellis 1968).
Specimen examined. On Meliolales on living leaves of Persea palustris,
U.S.A, Florida, Cove Springs, 1890, G. Martin, (IMI 16307).
Illustrations. This species was illustrated by Ellis (1968).
Notes. The specimen IMI 16307 was analysed, but no fungal cells were seen.
Spiropes guareicola (F. Stevens) Cif., Sydowia 9(1–6): 302, 1955
Fig. 12
≡ Helminthosporium guareicola F. Stevens [as ‘Helmisporium guareicolum’], Bot.
Gaz. 65(3): 241, 1918.
≡ Pleurophragmium guareicola (F. Stevens) S. Hughes, Can. J. Bot. 36: 797,
1958.
= Cladosporium elegans var. singaporense Sacc., Bull. Orto Bot. Regia Univ. Na-
poli 6: 60, 1921.
= Helminthosporium agellatum H.S. Yates [as ‘Helmisporium’], Philipp. J. Sci.
(Bot.) 13: 383, 1918.
= Helminthosporium spirotrichum Sacc. [as ‘Helmisporium’], Boll. Orto bot. 6:
61, 1921.
Description. Colonies effuse, dark brown to black, hairy. Hyphae supercial,
branched, septate, 2–4 µm wide, pale olivaceous-brown, smooth. Conidiophores
arising singly or in groups, as lateral branches on the hyphae, erect, sterile low-
er part straight or exuous, upper fertile part in zigzag shape, septate, up to
400 µm long, 6–9 µm thick, brown to dark brown, paler towards the apex, more
or less smooth, with numerous well-dened, dark conidial scars. Conidia soli-
tary, broadly fusiform, truncate at the base, with 3 to 5 pseudosepta, (25–)35–
52(–60) × (7–)8–10(–13) µm, 3.5–5 µm wide at the base, pale to dark brown or
olivaceous-brown, smooth as seen by SEM.
Specimen examined. On leaves of Cyrtophyllum fragrans (Gentianaceae),
Singapore, 1921, Baker (IMI 49160, type of Helminthosporium spirotrichum); on
Meliola sp. on leaves of Daniellia thurifera (Fabaceae), Sierra Leone, 1936, F.C.
Deightonii M1267 (IMI 10010).
Illustrations. This species was illustrated by Ellis (1968).

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Known hosts and distribution. On colonies of Asteridiella spp., Irenopsis spp.
and Meliola spp. on living leaves of various plants in Bougainville Islands, Gha-
na, India, Indonesia, Malaysia, Philippines, Puerto Rico, Sabah, Sierra Leone,
Solomon Islands and Uganda (Ellis 1968).
Notes. Spiropes guareicola is the type species of the genus Spiropes and it
differs from other species of the genus by the presence of zigzag-shaped con-
idiophores in the fertile upper parts (Ellis 1968). S. guareicola presents smooth
conidia, a feature that is only evident by SEM.
Spiropes helleri (F. Stevens) M.B. Ellis, Mycol. Pap. 114: 14, 1968
Fig. 13
≡ Helminthosporium helleri F. Stevens [as ‘Helmisporium], Bot. Gaz. 65(3): 242,
1918.
= Helminthosporium leucosykes H.S. Yates [as ‘Helmisporium leucosykeae’],
Philipp. J. Sci., C, Bot. 13(6): 382, 1918.
= Helminthosporium maculosum Sacc. [as ‘Helmisporium’], Atti Accad. Sci.
Ven.-Trent.-Istr. 10: 91, 1919 [1917].
≡ Pleurophragmium maculosum (Sacc.) S. Hughes, Can. J. Bot. 36: 797, 1958.
Figure 12. Spiropes guareicola (IMI 10010) a conidiophore with scars and a young conidium shown in optical section
bbase of a conidiophore growing on a hypha of Meliola sp. shown in optical section c conidia shown in optical section
(two drawings on the left-hand side) and as seen by SEM (two drawings on the right-hand side) d, e as seen by SEM
dzigzag-shaped conidiophore with scars e conidium. Scale bars: 5 μm (a–c); 8 μm (d); 10 μm (e).

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Description. Colonies effused, dark brown to black, hairy. Hyphae super-
cial, branched, septate, 1–3 µm wide, straw-coloured or pale brown, smooth.
Conidiophores arising singly as terminal or lateral branches on the hyphae,
erect, straight or exuous, septate, up to 600 µm long, 5–8 µm wide, brown
to dark brown, paler towards the apex, smooth, with scattered conidial scars.
Conidia solitary, obclavate, frequently rostrate, 3(–4)–septate, (26–)36–
43(–50) × (6–)7–10(–13) µm, 3–4 µm wide at the truncate base, pale brown to
brown, verruculose. As seen by SEM, the ornamentation of the spores is clearly
reticulated, with thin networks and no ridges.
Specimens examined. On Meliola sp. on leaves of Cupania guatemalensis
(Sapindaceae), Panama, Chiriquí Province, Botanical Garden of the Autono-
mous University of Chiriquí (UNACHI), 8°25'55"N, 82°27'03"W, 34 m a.s.l., 11
February 2020, M. A. Bermúdez-Cova, A. Sanjur MB92 (UCH15489, M); on Me-
liola sp. on living leaves of Pterocarpus santalinoides (Fabaceae), Benin, Atlan-
tique, Attogon, Niaouli Forest, 6°44'40"N, 2°7'53"E, 72 m a.s.l., 20 September
2022, A. Krauß, A. Tabé, O. Koukol, N.S. Yorou, AK15 (M).
Additional specimens examined. On Meliolales on living leaves of an unde-
termined plant, Gold Coast Colony, Banau, 1949, S.J. Hughes 1141 (IMI44564);
Figure 13. Spiropes helleri (IMI130940) a supercial hyphae growing on a colony of Meliola sp. on a leaf of Cupania gua-
temalensis b conidiophore growing on a hypha of Meliola sp. shown in optical section c conidia shown in optical section
(drawing on the left-hand side) and as seen by SEM (drawing on the right-hand side) d, e as seen by SEM d part of a
conidiophore with a scar e conidium. Scale bars: 1 mm (a); 5 μm (b); 6 μm (c); 4 μm (b); 5 μm (c).

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on Meliola sp. on leaves of Myrcia deexa, Puerto Rico, El Alto de la Bandera,
F.L. Stevens 8268 (IMI9991, type of Helminthosporium helleri).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Asteridiella spp., Irenopsis
spp. and Meliola spp. on living leaves of various plants in Ghana, Malaysia,
New Caledonia, Philippines, Puerto Rico, Sabah, Sierra Leone and Uganda (Ellis
1968). Spiropes helleri is reported here for the rst time for Benin and for main-
land America (Panama).
Notes. Spiropes helleri is similar to S. effusus, S. dorycarpus and S. leonensis
by the presence of obclavate to sometimes fusiform conidia, but differs from
the rst two by wider conidia (3.8–4.5 µm in S. effusus and 5–7 µm in S. dory-
carpus) and from the last one by narrower ones (10–11µm).
Spiropes intricatus (Sacc.) M.B. Ellis, Mycol. Pap. 114: 9, 1968
Fig. 14
≡ Brachysporium intricatum Sacc., Atti Accad. scient. Veneto-trent.-istriana, Ser.
3, 10: 88, 1919.
= Spiropes pirozynskii M.B. Ellis, Mycol. Pap. 114: 19, 1968. New synonym pro-
posed in this study.
Description. Colonies effuse, straw-coloured, olive or olivaceous-brown, velvety
or hairy. Hyphae supercial, branched, anastomosing, septate, 1–2 µm wide,
pale olivaceous brown, smooth. Conidiophores arising singly or in groups, ter-
minally or laterally from the hyphae, erect or ascending, straight or exuous,
septate, up to 900 µm long, 2–5 µm thick along most of their length, swollen
to 4–9 µm towards the apex and in intercalary parts that produce conidia, pale
olivaceous-brown to brown, reticulate as seen by SEM, with scattered cylindrical
scars. Conidia solitary, straight or slightly curved, oblong-ellipsoid or obovate to
clavate, truncate at the base, mostly 3–septate, (13–)16–23(–25) × (4.5–)6–
8µm, 1.5–3 µm wide at the base, the cells at each end of a conidium pale brown,
intermediate cells brown, ornamented. As seen by SEM, the ornamentation of the
spores is distinctly reticulated, with thin to thick networks that can form ridges.
Specimens examined. On Irenopsis sp. on Lindackeria bukobensis (Acha-
riaceae), Tanzania, Kigoma, 1964, K.A. Pirozynski M418 b&c (IMI 106645b-c,
type of Spiropes pirozynskii); on leaves of Camellia drupifera (Theaceae), Nepal,
Kathmandu, Godawari, 1986, U. Budathoki KU294 (IMI323287).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliolales on living leaves of
various plants in Ghana, Philippines and Tanzania (Ellis 1968).
Notes. Spiropes intricatus and S. deightonii are the only known species of
the genus that present conidiophores that swell in the areas where conidia are
formed (Figs 9, 14; Ellis (1968)). Spiropes deightonii differs from S. intricatus
by the presence of smaller conidia (12–14 µm long) that are more obovate or
clavate rather than oblong-ellipsoid. The type specimen of S. pirozynskii (IMI
106645b-c) is morphologically similar to S. intricatus. Both species present ob-
long-ellipsoid conidia with a similar size range (Fig. 15). Therefore, we propose
S. pirozynskii as a synonym of S. intricatus.

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Figure 14. Spiropes intricatus (IMI 106645b-c) a conidiophores, growing on a hypha of Irenopsis sp., shown in optical
section b conidia shown in optical section (the thickness of the wall is indicated only in the drawings on the upper row)
and as seen by SEM (second row right) c, d as seen by SEM c conidiophore with scars d conidium. Scale bars: 5 μm (a);
3 μm (b); 7 μm (c); 8 μm (c).
Spiropes japonicus (Henn.) M.B. Ellis, Mycol. Pap. 114: 22, 1968
Fig. 16
≡ Podosporium japonicum Henn., Bot. Jb. 29: 152, 1900.
= Helminthosporium insigne Gaillard ex Sacc. [as ‘Helmisporium’], Atti Accad.
Sci. Ven.-Trent.-Istr. 10: 89, 1917.
Description. Colonies effuse, amphigenous, sometimes dense, dark brown to
black, with tightly packed hyphae that form large, erect, dark synnemata clearly
visible under the stereomicroscope. Hyphae supercial, branched, septate, 1–4
µm wide, pale olivaceous-brown, smooth. Conidiophores tightly packed to form
dark brown to blackish synnemata up to 1 mm high, spreading out at the apex
and upper half of the synnemata; conidiophores individually exuous or straight,
thick-walled, septate, 6–8 µm thick, brown to dark brown at the base, paler to-
wards the apex, smooth, with scattered cylindrical scars. Conidia solitary, fusi-
form to obclavate, with 4(–6) pseudosepta, (50–)67–80 × (7–)8–14 µm, 2–3
µm wide at the apex, 3–5 µm at the truncate base, pale brown to brown, striate.
Specimens examined. On Meliola sp. on living leaves of Asteraceae, Pan-
ama, Chiriquí Province, Boquerón District, Chuspa Hydroelectric, 8°32'20"N,
82°36'21"W, 281 m a.s.l., 6 March 2020, M. A. Bermúdez-Cova, A. Sanjur, S.

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Figure 15. Scatter plot of spore size (width and length) of species of Spiropes.
Samaniego, MB120 (UCH15492); on Meliola sp. on living leaves of Fabace-
ae, Panama, Chiriquí Province, Bugaba District, area around Gariché River,
8°38'38.1"N, 82°41'19.6"W, 566 m a.s.l., 8 March 2020, M. A. Bermúdez-Cova,
A. Sanjur, A. Villarreal, MB123 (UCH15493, M).
Additional specimens examined. On Irenina entebbeensis on Alchornea hir-
tella (Euphorbiaceae), Sierra Leone, 1939, Makump, M1774 (IMI 38813); on As-
teridiella aucubae on Aucuba japonica (Garryaceae), Japan, Ise, 1899, P. Hen-
nings (IMI 130973, type of Podosporium japonicum).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliolales on living leaves of vari-
ous plants in the Cook Islands, Japan, Malaysia, Papua New Guinea and Sierra Le-
one (Ellis 1968). Spiropes japonicus is reported here for the rst time for Panama.
Notes. Spiropes japonicus is the only known synnematous species of
Spiropes that produces conidia with 4–6 pseudosepta, as well as synnemata
that splay out at the apex and upper half (Ellis 1968).
Spiropes leonensis M.B. Ellis, Mycol. Pap. 114: 15, 1968
Fig. 17
Description. Colonies effuse, grey to dark blackish-brown, hairy. Hyphae su-
percial, branched, septate, 2–6 µm wide, pale brown, smooth. Conidiophores

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Figure 16. Spiropes japonicus (MB120, 123) a synnemata growing on a colony of Meliola sp. b conidiophores with scars
and a young conidium, shown in optical section c a conidium shown in optical section (drawing on the left) and as seen
by SEM (drawing on the right) d, e as seen by SEM d conidiophore with a scar e conidium. Scale bars: 1 mm (a); 10 μm
(b, c); 3 μm (d); 9 μm (d).
arising singly, as terminal and lateral branches on the hyphae, erect, straight
or exuous, septate, up to 700 µm long, 8–12 µm thick, sometimes swollen to
16–17 µm at the base, dark brown to dark blackish-brown, paler towards the
apex, smooth, with scattered conidial scars. Conidia solitary, obclavate, ros-
trate, 3(–4)–septate, (38–)40–54(–63) × (8–)10–11(–13) µm, 4–6 µm wide
at the truncate base, pale brown to brown, verruculose. As seen by SEM, the
ornamentation of the spores is distinctly reticulated, with thin networks and no
ridges. It was not possible to see the scars by SEM.
Specimen examined. On Meliola garciniae on leaves of Pentadesma
butyracea (Clusiaceae), Sierra Leone, Rokupr, 1951, F.C. Deighton M3920 (IMI
46589b, holotype); on Meliola garciniae on Pentadesma butyracea, Sierra Le-
one, near Rokupr, 1939, F.C. Deighton (IMI 9992a, type of Spiropes leonensis).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliola garciniae on living
leaves of Pentadesma butyracea (Clusiaceae) in Sierra Leone (Ellis 1968).

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Notes. Spiropes leonensis is similar to S. helleri by the presence of rostrate,
obclavate, 3–septate conidia (Ellis 1968). However, conidia in S. helleri are
smaller (36–43 µm).
Spiropes melanoplaca (Berk. & M.A. Curtis) M.B. Ellis, Mycol. Pap. 114: 28, 1968
Fig. 18
= Arthrobotryum melanoplaca Berk. & M.A. Curtis, J. Linn. Soc. Bot. 10(46): 360,
1868.
≡ Podosporium melanoplaca (Berk. & M.A. Curtis) Cif., Sydowia 9(1–6): 310,
1955.
= Podosporium dialii Bat. [as ‘dialiumii’], Atas Inst. Micol. 1: 266, 1960. New
synonym proposed in this study.
≡ Spiropes dialii (Bat.) M.B. Ellis, Mycol. Pap. 114: 27, 1968. New synonym pro-
posed in this study.
= Arthrobotryum scoparium Henn., Hedwigia 43(6): 397, 1904. New synonym
proposed in this study.
Description. Colonies effuse, dark brown to black, hairy, with tightly packed
hyphae that form large, erect, dark synnemata clearly visible under the
Figure 17. Spiropes leonensis (IMI 46589b) a conidiophore with scars and a young conidium, shown in optical section
b part of a conidiophore growing on a hypha of Meliola sp., shown in optical section c conidia shown in optical section
(rst two drawings, from left to right) and as seen by SEM d conidium as seen by SEM. Scale bars: 8.5 μm (a–c); 7 μm (d).

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stereomicroscope. Hyphae supercial, branched, septate, 1.5–6 µm wide, pale
olivaceous, smooth. Conidiophores tightly packed to form dark brown to black-
ish synnemata up to 1.5 mm high, spreading out at the apex, 20–80 µm thick,
splaying out at the apex. Individual hyphae straight or exuous, cylindrical,
2–6µm thick along most of their length, 5–8 µm thick near the apex, with nu-
merous small scars that may overlap like scales. As evident by SEM, the scales
are produced by the peeling of the outer wall layers where the scars are located.
Conidia straight or curved, fusiform to obclavate, 3-septate, (30–)40–52(–68)×
(7–)9–11(–14) µm, with the two middle cells usually golden brown or brown,
warty and the cells at each end paler. As seen by SEM, the ornamentation of the
spores is distinctly reticulated, with thin to thick networks and no ridges.
Specimens examined. On Meliola mangiferae on living leaves of Mangifera
indica (Anacardiaceae), Panama, Chiriquí Province, Los Algarrobos, 8°31'05"N,
82°25'25"W, 168 m a.s.l., 20 January 2020, M. A. Bermúdez-Cova, MB81; same
fungal and plant host, Panama, Chiriquí Province, Universidad Autónoma de
Chiriquí (UNACHI), 8°25'57"N, 82°27'02"W, 37 m a.s.l., 23 January 2020, M. A. Ber-
múdez-Cova, MB85 (UCH15487); same fungal and plant host, Panama, Chiriquí
Province, Los Algarrobos, Majagua River Trail, 8°28'56"N, 82°24'47"W, 101 m a.s.l.,
23 January 2020, M. A. Bermúdez-Cova, MB89 (UCH15488, M); same fungal and
plant host, Panama, Chiriquí Province, Meseta de Chorcha, 8°24'19"N, 82°13'26"W,
94 m a.s.l., 16 February 2020, M. A. Bermúdez-Cova, A. Sanjur, MB101 (UCH);
Figure 18. Spiropes melanoplaca (MB81, MB119, IMI189570a) a, b synnemata growing on hyphae of Meliola mangiferae
on living leaves of Mangifera indica c conidiophores with scars and young conidia shown in optical section. The thick-
ness of the wall is only shown in the rst conidiophore, from left to right d conidia, shown in optical section (left-hand
drawing) and as seen by SEM (right-hand drawing) e, f as seen by SEM e parts of conidiophores with scars f conidium.
Scale bars: 1.5 mm (a); b); 0.9 mm (c); 8 μm (d); 7 μm (e); 8 μm (f).

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same fungal and plant host, Panama, Chiriquí Province, Boquerón District, Hidro-
eléctrica Chuspa, 8°33'37"N, 82°36'22"W, 331 m a.s.l., 6 March 2020, M. A. Bermú-
dez-Cova, A. Sanjur, S. Samaniego, MB119 (UCH15491); On Meliola sp. on living
leaves of Angylocalyx oligophyllus (Fabaceae), Benin, Attogon, Niaouli, Niaouli
Forest, 6°44'42"N, 2°7'50"E, 69 m a.s.l., 28 February 2022, M.A. Bermúdez-Cova, A.
Tabé, I. Agonglo, O.P. Agbani, M. Piepenbring, N.S. Yorou, MB173 (M); on Meliola
mangiferae on living leaves of Mangifera indica, Benin, Attogon, Niaouli, Niaouli
Forest, 6°44'44"N, 2°7'49"E, 65 m a.s.l., 28 February 2022, M.A. Bermúdez-Cova, A.
Tabé, I. Agonglo, O.P. Agbani, M. Piepenbring, N.S. Yorou, MB180 (M).
Additional specimens examined. On Meliola mangiferae on Mangifera indica,
Brunei, 1974, W.T.H. Peregrine (IMI189570a); on Meliola sp. on Psychotria sp.
(Rubiaceae), Cuba, 1879, C. Wright (IMI 105348 and IMI 105349, syntypes of
Arthrobotryum melanoplaca).
Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Meliolales, especially Meliola
spp., on living leaves of various plants in Brazil, Cuba, China, Dominican Republic,
Ghana, Guadalcanal, India, Malaysia, Peru, Philippines, Sierra Leone, Tanzania, Trin-
idad and Uganda (Ellis 1968; Zhao et al. 1996; Dubey and Moonnambeth 2013).
Spiropes melanoplaca is reported here for the rst time for Benin and Panama.
Notes. According to Ellis (1968), the main difference between Spiropes
melanoplaca and S. dialii is the range of spore width, with S. melanoplaca hav-
ing wider spores (9–14 µm wide) than S. dialii (7–9 µm wide). However, after
revision of several specimens and herbarium material from both species, we
noticed that the aspect of the colonies, morphological features (both as seen
in LM and by SEM) are similar between the species and both species present
conidia with a similar size range (Fig. 15). Therefore, we propose S. dialii as a
synonym of S. melanoplaca.
Spiropes palmetto (W.R. Gerard) M.B. Ellis, Mycol. Pap. 114: 16, 1968
Fig. 19
≡ Helminthosporium palmetto W.R. Gerard, Grevillea 17(83): 68, 1889.
≡ Pleurophragmium palmetto (W.R. Gerard) S. Hughes, Can. J. Bot. 36: 778,
1958.
Description. Colonies effuse, dark brown to black, hairy. Hyphae super-
cial, branched, anastomosing, septate, 1–4 µm wide, pale olivaceous-brown,
smooth. Conidiophores arising singly or in groups, as terminal and lateral
branches on the hyphae, erect, straight or exuous, septate, up to 400 µm long,
6–10 µm thick, dark brown, paler towards the apex, smooth, with scattered
conidial scars. Conidia solitary, obclavate to fusiform, rostrate, with 2 septa
delimiting a barrel-shaped central cell and often with an additional dark central
pseudoseptum, (27–)30–46 × (7–)9–12(–15) µm, 3–5 µm wide at the truncate
base, brown, middle cells pale brown, smooth as seen by LM and SEM.
Specimens examined. On Meliola sp. on leaves of Elaeis guineensis (Are-
caceae), Ghana, Apremodo, 1949, S.J. Hughes 534 (IMI 38617); on Meliola sp.
on leaves of Sabal palmetto (Arecaceae), U.S.A, Louisiana (IMI 10032, type of
Helminthosporium palmetto).

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Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Irenopsis spp. and Meliola spp.
on living leaves of various plants in Ghana, Malaysia, New Zealand, Puerto Rico,
Sierra Leone and the U.S.A. (Ellis 1968).
Notes. Spiropes palmetto can be easily recognised by the presence of co-
nidia with two septa that delimit a barrel-shaped central cell and with a dark
central pseudoseptum (Ellis 1968).
Spiropes penicillium (Speg.) M.B. Ellis, Mycol. Pap. 114: 23, 1968
Fig. 20
≡ Podosporium penicillium Speg., Boln. Acad. nac. Cienc. Córdoba 11: 618,
1889.
≡ Arthrobotryum penicillium (Speg.) F. Stevens, Bot. Gaz. 65: 238, 1918.
= Arthrobotryum strychni Henn., Hedwigia 43: 397, 1904.
≡ Podosporium strychni (Henn.) Cif., Sydowia 9: 311, 1955.
= Arthrobotryum glabroides F. Stevens, Bot. Gaz. 65: 237, 1918.
≡ Podosporium glabroides (F. Stevens) Cif., Sydowia 9: 309, 1955.
Figure 19. Spiropes palmetto (IMI 10032) a conidiophore growing on a hypha of Meliola sp., shown in optical section
bconidia shown in optical section. The thickness of the walls is only shown in the two last drawings c, d as seen by SEM
c part of a conidiophore with a scar d conidium. Scale bars: 7 μm (a); 5 μm (b); 6 μm (c); 7 μm (d).

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Description. Colonies effuse, yellowish to dark olivaceous-brown, velvety, with
tightly packed hyphae that form large, erect, dark synnemata clearly visible
under the stereomicroscope. A bright yellow pigment diffuses out when colo-
nies are mounted in lactic acid or lacto-phenol. Hyphae supercial, branched,
septate, 1–2 µm wide, yellowish, pale olive, smooth. Conidiophores tightly
packed to form dark brown to blackish synnemata up to 650 µm long, 10–
40µm thick, often splaying out to a width of 100 µm at the apex. Individual
hyphae straight or exuous, cylindrical, 1–2 µm thick near the base, 2–3.5µm
thick near the apex, pale olivaceous-brown, smooth, with numerous small
conidial scars. Conidia solitary, fusiform or occasionally almost cylindrical,
mostly 3(–5)–septate, 16–23(–37) × (3–)3.5–5(–7) µm, tapering to about
1µm at the apex and base, middle cells pale brown, the cells at each end
paler, surface wrinkled or verruculose. As seen by SEM, the ornamentation of
the spores is distinctly reticulated, with thin to thick networks that can form
ridges-like structures.
Specimen examined. On Meliola calva on leaves of Lauraceae, Brasil, S. Pau-
lo, Apiahy, 1881, J. Puiggari 1483 (IMI 131184, type of Podosporium penicilli-
um); on Meliola sp. on leaves of Oxyanthus sp. (Rybiaceae), Sierra Leone, 1951,
D.S. Rennis (IMI 51664).
Figure 20. Spiropes penicillium (IMI 51664) a conidiophores with scars (the thickness of the wall is shown on the
right-handed drawing) b conidia shown in optical section (rst two left-hand drawings) and as seen by SEM c, d as seen
by SEM c tips of conidiophores with scars d conidia. Scale bars: 5 μm (a); 2.5 μm (b); 3 μm (c); 5 μm (d).

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Illustrations. This species was illustrated by Ellis (1968).
Known hosts and distribution. On colonies of Asteridiella spp. and Meliola
spp. on living leaves of various plants in Brazil, Congo, Costa Rica, Ghana, Ivory
Coast, Nigeria, Sierra Leone and Uganda (Ellis 1968).
Notes. Spiropes penicillium is easily distinguishable from other known
synnematous species of the genus Spiropes by the presence of fusiform to
cylindrical conidia without rostra. In addition, a bright yellow pigment diffuses
out of the cells when colonies are mounted in lactic acid or lacto-phenol (El-
lis 1968).
Key to species of Atractilina and Spiropes hyperparasitic on Meliolales
1 Conidiophores synnematous ........................................................................2
– Conidiophores single or in groups ...............................................................7
2 Synnemata straw-coloured to pale olivaceous; conidiophores with dentic-
ulate conidiogenous loci; pale multiseptate conidia ................A. parasitica
– Synnemata dark brown to black; conidiophores with cicatrised conidioge-
nous loci; conidia pigmented and multiseptate ..........................................3
3 Synnemata up to 400 μm long; conidia mostly crescent shape .................
.......................................................................................... S. croissantiformis
– Synnemata longer, from 700 μm to 1.5 mm long; conidia fusiform to ob-
clavate, occasionally cylindrical ...................................................................4
4 Conidia fusiform to almost cylindrical; a yellow pigment diffuses out when
colonies are mounted in lactic acid or lacto-phenol .............. S. penicillium
– Conidia fusiform to obclavate; no yellow pigment ......................................5
5 Conidia always 4–6 septate ...................................................... S. japonicus
– Conidia always 3–septate ............................................................................6
6 Conidia 17–25 × 5–6.5 μm ...........................................................S. clavatus
– Conidia 40–52 × 9–11 μm .................................................... S. melanoplaca
7 Conidia with 3–6 pseudosepta.....................................................................8
– Conidia 1–3–septate ..................................................................................10
8 Conidiophores in larger groups; conidia with 3–6 (usually 4 or 5) pseu-
dosepta .........................................................................................S. capensis
– Conidiophores single or in small groups; conidia with 3–5 pseudosepta ....9
9 Conidiophores with zigzag shape; conidia with 3–5 pseudosepta, fusi-
form to obclavate ...................................................................... S. guareicola
– Conidiophores without zigzag shape; conidia with 3–4 pseudosepta, obo-
vate ................................................................................................ S. fumosus
10 Conidia 1–septate .......................................................................................11
– Conidia 3–septate .......................................................................................12
11 Conidia obpyriform, verrucose .................................................S. armatellae
– Conidia obpyriform, smooth .................................................S. armatellicola
12 Conidia oblong-ellipsoid .............................................................S. intricatus
– Conidia of various shapes, not oblong-ellipsoid .......................................13
13 Conidia obovate to clavate; conidiophores swollen towards the apex or in
areas where conidia are produced ............................................S. deightonii
– Conidia ovate or fusiform to obclavate; conidiophores not swollen to-
wards the apex or in areas where conidia are produced ..........................14

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14 Conidia obclavate; central cells barrel-shaped ..........................................15
– Conidia ovate or fusiform to obclavate; without central barrel-shaped
cells ..............................................................................................................16
15 Conidia with 3 true septa ......................................................... S. caribensis
– Conidia with 2 septa and a dark central pseudoseptum ...........S. palmetto
16 Conidia ovate ........................................................................... S. carpolobiae
– Conidia fusiform to obclavate ....................................................................17
17 Conidia 3–4.5 μm wide .................................................................. S. effusus
– Conidia wider ...............................................................................................18
18 Conidia 17–25 μm long .......................................................S. angylocalycis
– Conidia longer ..............................................................................................19
19 Conidia 20–35 μm long ...........................................................S. dorycarpus
– Conidia longer ..............................................................................................20
20 Conidia 36–48 μm long ................................................................... S. helleri
– Conidia 40–54 μm long ..............................................................S. leonensis
In Fig. 21, we propose a visual key to the known species of Spiropes hyper-
parasitic on Meliolales.
Figure 21. Visual key to known species of Spiropes hyperparasitic on Meliolales.

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Molecular position of species of Atractilina and Spiropes
In order to know the systematic positions of species of Atractilina and Spiropes
hyperparasitic on Meliolales, new sequences of recently-collected specimens
were obtained.
Figure 22. Phylogenetic tree inferred from a Maximum Likelihood analysis of nuc LSU rDNA sequences of members of
the Dothideomycetes, including new sequences of Atractilina parasitica and Malacaria meliolicola (written with bold let-
ters). The tree is rooted with sequences of species of the orders Capnodiales and Mycosphaerellales. Bootstrap values
are indicated above the branches. Sequences downloaded from GenBank are given with accession numbers.

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The BLAST query revealed that the nrLSU sequences of Atractilina parasit-
ica (specimens MB136 and MB178) show approximately 82% similarity with
sequences of species of the Dothideomycetes, such as Botryosphaeria spp.,
Helminthosporium asterinum Cooke, Hysterobrevium mori (Schwein.) E. Boehm
& C.L. Schoch and Neoheleiosa lincangensis Mortimer, amongst others. In the
tree inferred from the analysis of LSU sequences of 45 specimens of sever-
al orders of Dothideomycetes (Fig. 22), the sequences of A. parasitica are lo-
cated in a well-supported clade that comprises species of Pleosporales, such
as Ellismarsporium parvum R.F. Castañeda & W.B. Kendr., Kirschsteiniothelia
aethiops (Sacc.) D. Hawksw. and Helminthosporium asterinum. In addition, the
sequences of A. parasitica cluster together in a strongly-supported clade with
two DNA sequences we obtained from Malacaria meliolicola (specimens AK4H
and AK06H), a hyperparasitic perithecioid fungus that usually grows amongst
the synnemata of A. parasitica on coffee leaves (see Bermúdez-Cova et al.
(2023b) for the updated species description of M. meliolicola).
As for species of Spiropes, the BLAST query revealed that the nrITS sequenc-
es of Spiropes melanoplaca (specimens MB81 and MB119) and Spiropes japon-
icus (specimen MB 120) are not closely related to each other (60% similarity)
and show between 88 and 90% similarity with species of the Leotiomycetes,
such as Lophodermium actinothyrium Fuckel and Hypoderma spp., amongst
others. Placement on to the Pezizomycotina tree version 2 in T-BAS conrmed
that the newly-generated ITS sequences for the two species of Spiropes are
placed in the Leotiomycetes (Fig. 23).
Discussion
Atractilina and Spiropes, two genera with heterogeneous species
Morphology-based identication of a species can be very dicult, especially
amongst asexual or non-sporulating fungi (Jeewon et al. 2002; Promputtha et
al. 2005, 2007). However, it continues to be an essential tool, especially for
understudied groups of fungi and when DNA sequences are not available or
scarce (Raja et al. 2017). The morphological analyses and the literature review
of specimens of Atractilina and Spiropes revealed that both genera include
highly heterogeneous species that are not necessarily congeneric with the type
species of each genus.
The type species of Atractilina, Atractilina callicarpae Dearn. & Barthol.
(= Atractilina parasitica (G. Winter) Deighton & Piroz.), has consistently true
synnematous conidiophores, denticulate conidiogenous loci, pale pluriseptate
(phragmoseptate) conidia and a hyperparasitic lifestyle (Deighton and Pirozyns-
ki 1972; Mel’nik and Braun 2013). Based on these characteristics, only three
species of the genus are congeneric with A. parasitica, namely A. alinae Melnik
& U. Braun, A. biseptata R.F. Castañeda and A. calycini T.K. Jana, S.N. Ghosh &
A.K. Das (Castañeda-Ruiz 1986; Jana et al. 2006; Mel’nik and Braun 2013). The
remaining two species present non-synnematous conidiophores and are prob-
ably not congeneric. Atractilina asterinae (Hansf.) Deighton & Piroz. is a species
hyperparasitic on Asterinales and presents single conidiophores and distosep-
tate conidia (Deighton and Pirozynski 1972). Atractilina hymenaeae Bat. & J.L.
Bezerra (introduced as Atractina hymenaeae by the authors) is hyperparasitic

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Figure 23. Placement of Spiropes japonicus and S. melanoplaca on to Pezizomycotina reference tree version 2 in T-Bas.
Only the Leotiomycetes clade is shown. The tree is the result of RAxML analysis of nuc ITS rDNA with 500 bootstraps
replicates. For each node, the Maximum Likelihood bootstrap (≥ 70%) is presented as thick branches. Names of Spiropes
species with newly-generated sequence data are written in bold.

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on Meliolales, but also with non-synnematous conidiophores and conidia with
a variable number of septa (Batista and Bezerra 1961). Therefore, we believe
that both species have been incorrectly assigned to the genus Atractilina.
The description of A. parasitica introduced by Deighton and Pirozynski
(1972) is very broad. As a result, specimens with signicant morphological
variations are grouped into a single species concept. For example, Chen and
Tzean (2007) described a parasitic fungus from Taiwan growing on decaying
leaves of Liquidambar sp. (Altingiaceae), with conidia that resemble those of
A. parasitica. However, conidiophores of this fungus are non-synnematous and
very short (less than 15 μm long), a feature that has never been reported before
for A. parasitica. It is necessary to re-evaluate this and other identications, to
narrow the species concept of A. parasitica, as well as to complement it with
DNA sequence data.
The DNA molecular analyses of the nrLSU rDNA region of the specimens of
A. parasitica from Benin revealed that this species belongs to the Dothideomy-
cetes. The Dothideomycetes are the largest and most diverse class of fungi and
comprise species that exhibit a broad range of lifestyles, including saprotrophs,
plant pathogens, mycoparasites and hyperparasites, as well as lichenised and
lichenicolous fungi (Pem et al. 2021). They typically produce ask-like struc-
tures called pseudothecia, though apothecial, hysterothecial and cleistothe-
cioid ascomata also exist (Hessen and Jahns 1973; Valenzuela-Lopez et al.
2019). Bitunicate asci are one of the diagnostic characters for Dothideomy-
cetes taxonomy (Von Arx and Müller 1975; Pem et al. 2021). Asexual stag-
es are frequent amongst pathogenic genera in the families Cladosporiaceae,
Mycopsphaerellaceae, Pleosporaceae and Tubeuaceae, amongst others (Hyde
et al. 2013; Wanasinghe et al. 2018; Hongsanan et al. 2020). Conidiophores
in these anamorphic species are usually solitary or in groups forming synne-
mata (Thambugala et al. 2017). The sequences of A. parasitica showed 98%
similarity with sequences of Malacaria meliolicola (Dothideomycetes, Ascomy-
cota), a pseudothecioid hyperparasite that was found repeatedly amongst the
synnemata of A. parasitica (Bermúdez-Cova et al. 2023b). The pseudothecia of
M. meliolicola were also found to be growing without the presence of synnema-
ta of A. parasitica. These colonies were used to extract the DNA of M. melioli-
cola. Therefore, the systematic position of A. parasitica in the Dothideomycetes
and the anamorph-teleomorph connection between these two species are con-
rmed. This connection has been proposed in the past for these fungi on leaves
of Coffea arabica (Hansford 1941, 1946; Bermúdez-Cova et al. 2023b). Here,
a DNA sequence from a specimen of A. parasitica on Meliola sp. on leaves
of Clerodendrum capitatum clustered with the aforementioned sequences in
a highly-supported clade. The phylogenetic analysis of the nrLSU DNA locus
showed that sequences of A. parasitica are located in a well-supported sub-
clade together with other species of Pleosporales s.l., such as Ellismarsporium
parvum (Zhang et al. 2020). Many species of the Dothideomycetes, especially
the asexual genera, are known to be polyphyletic (Schoch et al. 2009). To con-
rm the systematic hypothesis and to determine the placement of A. parasitica
at family level, the use of multi-loci phylogenies is necessary in the future.
As for the genus Spiropes, the generic diagnosis given by Ellis (1968, 1971)
allows us to include in this genus all species with cicatrised conidiogenous
cells and conspicuous, at and numerous scars, as well as pigmented, mostly

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obclavate phragmoconidia with 1–9 septa or pseudosepta. Seifert and Hughes
(2000) proposed an amendment of this generic concept to also include species
with dictyoconidia. As a result, S. dictyosporus is the only known species of the
genus with muriform conidia. However, this morphological diagnosis allows
for species with a wide range of types of conidiophores, conidiogenesis and
conidia to be included in Spiropes (McTaggart et al. 2007). For example, the
type species of the genus, Spiropes guareicola (F. Stevens) Cif., has distinctly
sympodial-geniculate (zigzag-shaped) conidiophores, a character that is not
present in any other known species of the genus (Ellis 1968). This species,
in addition, presents distoseptate conidia, i.e. conidia with pseudosepta, a
morphological feature that is present only in four species, namely S. capen-
sis, S. fumosus, S. guareicola and S. japonicus. The remaining species of the
genus present euseptate conidia (Ellis 1968, 1971). It is also possible to nd a
wide range of conidial shapes, such as obpyriform, obovate, ovate and oblong
ellipsoid, to obclavate and fusiform (see the visual key to species of Spiropes
in Fig. 21). Therefore, Spiropes is currently a genus with morphologically highly
heterogeneous species and probably polyphyletic.
Identifying species of Spiropes, based on morphology alone, is not always
easy. The most comprehensive key to species of the genus was proposed by El-
lis (1968). However, this key is mainly based on the differences in the size range
of the conidia of the species and, in some cases, these size differences are very
subtle. Particular attention should be paid to herbarium specimens, as they may
include immature or not well-preserved spores that can affect measurement
results (Ordynets et al. 2021). We believe that other morphological character-
istics that are not visible using standard light microscopy techniques should
be considered when identifying species of Spiropes (e.g. Lutzoni et al. (2004)).
Scanning electron Microscopy (SEM), for example, allowed us to observe for the
rst time the surface of the conidia of species of Spiropes. Spiropes diallii and
S. melanoplaca were considered as different species by Ellis (1968). However,
both species have overlapping spore-size ranges and the morphological anal-
ysis by SEM revealed that these species also have similar conidiogenesis and
ornamentation patterns on conidia. This situation is similar for S. intricatus and
S. pirozynskii. Therefore, we propose both groups of species as synonyms.
As for the molecular-based identication of species of Spiropes, there are cur-
rently no DNA sequences available in publicly-accessible databases. Species of
the genus remain “incertae sedis” for many taxonomic ranks and it is dicult to
assign new DNA sequences to species concepts (Bermúdez-Cova et al. 2022,
2023a). The DNA sequences generated for the rst time in the context of this study
suggest that species of Spiropes hyperparasitic on Meliolales may be polyphyletic
in the Leotiomycetes. Fungi in the class Leotiomycetes are ecologically diverse
and have been described as aquatic hyphomycetes, ectomycorrhizal parasites,
endophytes, fungal parasites, mycorrhizal fungi, nematode-trapping fungi and
plant-pathogens, amongst others (Wang et al. 2006a; Johnston et al. 2019). Many
fungi have been suggested to belong to this class without any clear teleomorphic
connection (Wang et al. 2006b). Up to date, no sexual stages have been linked
to any species of Spiropes (Bermúdez-Cova et al. 2022). There is one genus with
species morphologically similar to species of Spiropes, namely Pseudospiropes
M.B. Ellis (Helotiales, Leotiomycetes; Ellis (1971)). Species of this genus differ
from species of Spiropes by broadly enlarged, thickened, protuberant, strongly

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melanised conidiogenous loci and distoseptate conidia only (Castañeda-Ruiz et
al. 2001; McTaggart et al. 2007). Species of Pseudospiropes have Strossmayeria
Schulzer (Helotiales, Leotiomycetes) teleomorphs (Iturriaga and Korf 1984, 1990;
Castañeda-Ruiz et al. 2001). Thus, there is a possibility that species of the genus
Spiropes also belong to the Leotiomycetes. It is necessary to continue generating
new DNA sequences from the different species of the genus in order to conrm
this hypothesis, especially from those species that form part of mixed infections.
It is dicult to obtain molecular sequence data from hyperparasites especially
because of the fact that they develop intermingled with the primary parasite and
many other organisms and, as a result, no specic set of molecular methods
has been developed to study hyperparasites (Bermúdez-Cova et al. 2022; Bermú-
dez-Cova et al. 2023a). As a consequence, isolating and sequencing hyperpara-
sitic fungi is a challenging task. There is also a lack of sequences of hyperpara-
sitic fungi in public. Therefore, the sequences obtained can be related to existing
species concepts only based on morphology databases (Bermúdez-Cova et al.
2023b). For hyperparasitic fungi on Meliolales, for example, it is advised to obtain
the same or very similar DNA sequences repeatedly from a given morphospecies
in order to be sure to have the correct DNA sequence of that morphospecies.
Despite many attempts, it was not possible to obtain DNA sequences from some
of the species included in this study. However, this research provides valuable
information that lays the foundation for future research on hyperparasites in Me-
liolales, highlighting the importance of eld work paired with molecular for the
study of challenging fungal groups. Further methodologies, such as metabar-
coding, could represent another way to try to isolate the DNA of these organisms.
The need for re-evaluation, resampling and epitypication
Applications of names based on morphological characteristics without DNA
data is a challenge, resulting in the description of an excessive number of
species or, in contrast, in the overlooking of cryptic species that can only be
detected through molecular analyses (Hibbett et al. 2007; Crous et al. 2014;
Jayasiri et al. 2015). The knowledge of morphological characteristics, however,
is important to understand the evolution of fungal diversity (Raja et al. 2017).
Instead of describing new species as part of Atractilina and Spiropes, a re-eval-
uation of the natural concepts of both genera is needed. Here we propose a list
of actions that are necessary to carry out such a re-evaluation:
• Restudy the type species of each genus. When the type specimens of the
type species are not in good condition or there is no more fungal material
available for examination, it is necessary to recollect them. Epitypes and
neotypes should be designated in these cases.
• After redening the type species, all species belonging to the two genera
need to be recollected, re-analysed morphologically and compared to the
type species.
• The DNA of all existing species should be extracted, amplied and se-
quenced, in order to conrm or propose new concepts of genera and
species. Multi-loci phylogenetic analyses are necessary to validate or pro-
pose new systematic hypotheses.

207
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Atractilina and Spiropes are currently two repository genera of highly hetero-
geneous species and they may be split in the future, once species and genus
concepts are validated respectively by morphology and molecular methods.
Acknowledgements
We are grateful to the University of Parakou and the University of Abom-
ey-Calavi, Benin, for the support and facilities made available for this study.
We acknowledge help by Dr. Pierre Agbani (Botanical Garden of the Université
d’Abomey-Calavi) for his assistance with the identication of host plants and
help by Daouda Dongnima during eldwork. Special thanks to Affoussatou
Tabé, Alicia Sanjur and Anna Krauss for their support and collecting efforts
throughout the whole research process. We acknowledge the support and fa-
cilities made available by Orlando Cáceres and the Universidad Autónoma de
Chiriquí (UNACHI) in Panama. The Environmental Ministry of Panama (MiAm-
biente) is thanked for issuing the collection and export permits (SE/APHO-1-
2019, SEX/H-5-2020, PA-01-ARG-049-2021). We are grateful to the Ministry of
Environment of the Benin Republic for issuing the collecting permits and for the
elaboration of the ABS Nagoya Protocol documents n° 636/DGEFC/ANC-APA/
DCPRN/PF-APA.
Additional information
Conict of interest
The authors have declared that no competing interests exist.
Ethical statement
No ethical statement was reported.
Funding
The rst author acknowledges support from the German Academic Exchange Service
(DAAD), within the framework of the scholarship programme for doctoral studies in Ger-
many (Ref. no.: 91726217).
Author contributions
MB-C compiled and analyzed the data and wrote the rst draft of the manuscript. MB-C,
TH, NY and MP contributed to writing and editing the manuscript. All authors contribut-
ed to the article and approved the submitted version.
Author ORCIDs
Miguel A. Bermúdez-Cova https://orcid.org/0000-0002-7712-7347
Tina A. Hofmann https://orcid.org/0000-0003-1124-402X
Nourou S. Yorou https://orcid.org/0000-0001-6997-811X
Meike Piepenbring https://orcid.org/0000-0002-7043-5769
Data availability
All of the data that support the ndings of this study are available in the main text or
Supplementary Information.

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Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
References
Abarca GH, Ruiz RC, Arias RM, Saikawa M, Stadler M (2007) Anamorphic fungi from
submerged plant material: Acumispora verruculosa, Pleurophragmium aquaticum and
P. miniumbonatum. Mycotaxon 101: 89–97.
Akoègninou A, Van der Burg WJ, Van der Maesen LJG (2006) Flore Analytique du Bénin.
Backhuys Publishers, Wageningen, 1034 pp.
Alcorn JL (1988) The taxonomy of “Helminthosporium” species. Annual Review of Phy-
topathology 26(1): 37–56. https://doi.org/10.1146/annurev.py.26.090188.000345
Bánki O, Roskov Y, Döring M, Ower G, Vandepitte L, Hobern D, Remsen D, Schalk P, DeWalt
RE, Keping M, Miller J, Orrell T, Aalbu R, Abbott J, Adlard R, Adriaenssens EM, Aedo
C, Aescht E, Akkari N, et al. (2023) Catalogue of Life Checklist [Version 2023-07-10].
Catalogue of Life. https://doi.org/10.48580/dfry
Batista AC, Bezerra JL (1961) Novos ou raros Deuteromycetes. Memórias da Sociedade
Broteriana 14: 73–82.
Benson DA, Clark K, Karsch-Mizrachi I, Lipmanbn DJ, Ostell J, Sayers EW (2014) GenBank.
Nucleic Acids Research 42(D1): D32–D37. https://doi.org/10.1093/nar/gkt1030
Bermúdez-Cova MA, Cruz-Laufer AJ, Piepenbring M (2022) Hyperparasitic fungi on
black mildews (Meliolales, Ascomycota): Hidden fungal diversity in the tropics. Fron-
tiers in Fungal Biology 3: 885279. https://doi.org/10.3389/ffunb.2022.885279
Bermúdez-Cova MA, Haelewaters D, de Bekker C, Piepenbring M, Schoutteten N, Quandt
CA (2023a) Hyperparasitic fungi – denitions, diversity, ecology, and research. Autho-
rea, June 27, 2023. https://doi.org/10.22541/au.168787020.07281183/v1
Bermúdez-Cova MA, Krauß A, Sanjur A, Tabé A, Hofmann TA, Yorou NS, Piepenbring M
(2023b) Diversity of hyperparasitic fungi on Meliolales (Sordariomycetes, Ascomyco-
ta): New species, records, and molecular data from Benin and Panama. Mycological
Progress 22(9): 65. https://doi.org/10.1007/s11557-023-01913-5
Carbone I, White JB, Miadlikowska J, Arnold AE, Miller MA, Magain N, U’Ren JM, Lutzoni
F (2019) T-BAS version 2.1: Tree-Based Alignment Selector toolkit for evolutionary
placement of DNA sequences and viewing alignments and specimen metadata on
curated and custom trees. Microbiology Resource Announcements 8(29): e00328–
e19.https://doi.org/10.1128/MRA.00328-19
Castañeda-Ruiz RF (1986) Fungi Cubenses, 20 pp.
Castañeda-Ruiz RF, Heredia G, Reyes M, Arias RM (2001) A revision of the genus Pseu-
dospiropes and some new taxa. Cryptogamie. Mycologie 22(1): 3–18. https://doi.
org/10.1016/S0181-1584(01)01057-0
Chen J-L, Tzean S-S (2007) Hyphomycetes from Taiwan. Akanthomyces and allied spe-
cies. Fungal Science 22(3, 4): 63–77.
Ciferri R (1955) Observations on meliolicolous Hyphales from Santo Domingo. Sydowia
9(1–6): 296–335.
Condit R, Pérez R, Daguerre N (2011) Trees of Panama and Costa Rica. Princeton Univer-
sity Press, Princeton NJ, 481 pp. https://doi.org/10.1515/9781400836178
Costantin JN (1888) Les Mucédinées Simples. Histoire, Classication, Culture et Rôle
des Champignons Inférieurs dans les Maladies des Végétaux et des Animaux. Klinck-
sieck, Paris, i-viii, 210 pp.[, 190 gs]
Crous PW, Giraldo A, Hawksworth DL, Rober V, Kirk PM, Guarro J, Robbertse B, Schoch
CL, Damm U, Trakunyingcharoen T, Groenewald JZ (2014) The Genera of Fungi: Fixing
the application of type species of generic names. IMA Fungus 5(1): 141–160. https://
doi.org/10.5598/imafungus.2014.05.01.14

209
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
De Beer ZW, Seifert KA, Wingeld MJ (2013) A nomenclator for ophiostomatoid genera and
species in the Ophiostomatales and Microascales. Biodiversity Series 12: 245–322.
Deighton FC, Pirozynski KA (1972) Microfungi. V. More hyperparasitic hyphomycetes.
Mycological Papers 128: 110.
Dubey R, Moonnambeth NA (2013) Hyperparasitism of Isthmospora spinosa Stevens
and Spiropes melanoplaca (Berk. & Curtis) Ellis on Meliola tylophorae – indicae Hos-
ag. parasitizing Tylophora indica (Burm. f.) Merill from India- A New Record. Journal
on New Biological Reports 2(1): 64–66.
Ellis MB (1968) Dematiaceous Hyphomycetes. IX. Spiropes and Pleurophragmium. My-
cological Papers 114: 1–44.
Ellis MB (1971) Dematiaceous Hyphomycetes. X. Mycological Papers 125: 1–30.
https://doi.org/10.1079/9780851986180.0000
Ellis MB (1976) More Dematiaceous Hyphomycetes. Kew, Commonwealth Mycological
Institute, 507 pp. https://doi.org/10.1079/9780851983653.0000
Fazekas AJ, Kuzmina ML, Newmaster SG, Hollingsworth PM (2012) DNA barcoding
methods for land plants. In: Kress WJ, Erickson DL (Eds) DNA Barcodes. Methods
in Molecular Biology. Humana, Totowa, 223–252. https://doi.org/10.1007/978-1-
61779-591-6_11
Hansford CG (1941) Contribution towards the fungus ora of Uganda. III. Proceedings of
the Linnean Society of London 153: 4–52. https://doi.org/10.1111/j.1095-8312.1941.
tb01378.x
Hansford CG (1946) The foliicolous ascomycetes, their parasites and associated fungi.
Mycological Papers 15.
Hessen A, Jahns HM (1973) ‘1974’ lichens. Eine Einführung in die Flechtenkunde.
Thiene, Stuttgart.
Hibbett DS, Binder M, Bischoff JF, Blackwell M, Cannon PF, Eriksson OE, Huhndorf S,
James T, Kirk PM, Lücking R, Lumbsch HT, Lutzoni F, Mathenya PB, McLaughlin DJ,
Powell MJ, Redhead S, Schoch CL, Spatafora JW, Stalpers JA, Vilgalys R, Aime MC,
Aptroot A, Bauer R, Begerow D, Benny GL, Castlebury LA, Crous PW, Dair YC, Gams W,
Geisers DM (2007) A higher-level phylogenetic classication of the fungi. Mycologi-
cal Research 111(5): 509–547. https://doi.org/10.1016/j.mycres.2007.03.004
Hofmann TA, Kirschner R, Piepenbring M (2010) Phylogenetic relationships and new
records of Asterinaceae (Dothideomycetes) from Panama. Fungal Diversity 43(1):
39–53. https://doi.org/10.1007/s13225-010-0042-4
Holubová-Jechová V, Sierra AM (1984) Studies on hyphomycetes from Cuba II. Hypho-
mycetes from the Isla de la Juventudeská. Česká Mykologie 38: 96–120.
Hongsanan S, Tian Q, Persoh D, Zeng X-Y, Hyde KD, Chomnunti P, Boonmee S, Bahkali AH,
Wen T-C (2015) Meliolales. Fungal Diversity 74(1): 91–141. https://doi.org/10.1007/
s13225-015-0344-7
Hongsanan S, Hyde KD, Phookamsak R, Wanasinghe DN, McKenzie EHC, Sarma VV, Boon-
mee S, Lücking R, Bhat DJ, Liu NG, Tennakoon DS, Pem D, Karunarathna A, Jiang SH,
Jones EBG, Phillips AJL, Manawasinghe IS, Tibpromma S, Jayasiri SC, Sandamali DS,
Jayawardena RS, Wijayawardene NN, Ekanayaka AH, Jeewon R, Lu YZ, Dissanayake
AJ, Zeng XY, Luo ZL, Tian Q, Phukhamsakda C, Thambugala KM, Dai DQ, Chethana
KWT, Samarakoon MC, Ertz D, Bao DF, Doilom M, Liu JK (2020) Rened families of
Dothideomycetes: Dothideomycetidae and Pleosporomycetidae. Mycosphere : Journal
of Fungal Biology 11(1): 1553–2107. https://doi.org/10.5943/mycosphere/11/1/13
Hosagoudar VB (2003) Armatellaceae, a new family segregated from the Meliolaceae.
Sydowia 55: 162–167.

210
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Hosagoudar VB, Biju CK, Abraham TK, Agarwal DK (2002) Spiropes armatellicola sp.
nov. from Kerala, India. Journal of Economic and Taxonomic Botany 26(3): 603–604.
Hyde KD, Jones E, Liu JK, Ariyawansa H, Boehm E, Boonmee S, Braun U, Chomnunti P,
Crous PW, Dai D-Q, Diederich P, Dissanayake A, Doilom M, Doveri F, Hongsanan S,
Jayawardena R, Lawrey JD, Li Y-M, Liu Y-X, Lücking R, Monkai J, Muggia L, Nelsen MP,
Pang K-L, Phookamsak R, Senanayake IC, Shearer CA, Suetrong S, Tanaka K, Thambu-
gala KM, Wijayawardene NN, Wikee S, Wu H-X, Zhang Y, Aguirre-Hudson B, Alias SA,
Aptroot A, Bahkali AH, Bezerra JL, Bhat DJ, Camporesi E, Chukeatirote E, Gueidan C,
Hawksworth DL, Hirayama K, De Hoog S, Kang J-C, Knudsen K, Li W-J, Li X-H, Liu Z-Y,
Mapook A, McKenzie EHC, Miller AN, Mortimer PE, Phillips AJL, Raja HA, Scheuer C,
Schumm F, Taylor JE, Tian Q, Tibpromma S, Wanasinghe DN, Wang Y, Xu J-C, Yacha-
roen S, Yan J-Y, Zhang M (2013) Families of Dothideomycetes. Fungal Diversity 63(1):
1–313. https://doi.org/10.1007/s13225-013-0263-4
Iturriaga T, Korf RP (1984) Studies in the genus Strossmayeria (Helotiales). 1. generic
delimitation. 2. Two lost species. 3. Three excluded taxa. Mycotaxon 20: 169–178.
Iturriaga T, Korf RP (1990) A monograph of the Discomycete genus Strossmayeria
(Leotiaceae), with comments on its anamorph, Pseudospiropes (Dematiaceae).
Mycotaxon 36: 383–454.
Jana TK, Das AK, Ghosh SN (2006) Studies on foliicolous fungi – I. Geobios (Jodhpur)
33(1): 9–16.
Jayasiri SC, Hyde KD, Ariyawansa HA, Bhat J, Buyck B, Cai L, Dai Y-C, Abd-Elsalam KA,
Ertz D, Hidayat I, Jeewon R, Jones EBG, Bahkali AH, Karunarathna SC, Liu J-K, Luang-
sa-ard JJ, Lumbsch HT, Maharachchikumbura SSN, McKenzie EHC, Moncalvo J-M,
Ghobad-Nejhad M, Nilsson H, Pang K-L, Pereira OL, Phillips AJL, Raspé O, Rollins AW,
Romero AI, Etayo J, Selçuk F, Stephenson SL, Suetrong S, Taylor JE, Tsui CKM, Vizz-
ini A, Abdel-Wahab MA, Wen T-C, Boonmee S, Dai DQ, Daranagama DA, Dissanay-
ake AJ, Ekanayaka AH, Fryar SC, Hongsanan S, Jayawardena RS, Li W-J, Perera RH,
Phookamsak R, de Silva NI, Thambugala KM, Tian Q, Wijayawardene NN, Zhao R-L,
Zhao Q, Kang J-C, Promputtha I (2015) The Faces of Fungi database: Fungal names
linked with morphology, phylogeny and human impacts. Fungal Diversity 74(1): 3–18.
https://doi.org/10.1007/s13225-015-0351-8
Jayawardena RS, Hyde KD, Chen YJ, Papp V, Palla B, Papp D, et al. (2020) One stop shop IV:
taxonomic update with molecular phylogeny for important phytopathogenic genera: 76–
100 (2020). Fungal Diversity 103: 87–218. https://doi.org/10.1007/s13225-020-00460-8
Jeewon R, Liew ECY, Hyde KD (2002) Phylogenetic relationships of Pestalotiopsis and
allied genera inferred from ribosomal DNA sequences and morphological characters.
Molecular Phylogenetics and Evolution 25(3): 378–392. https://doi.org/10.1016/
S1055-7903(02)00422-0
Johnston PR, Quijada L, Smith CA, Baral HO, Hosoya T, Baschien C, Pärtel K, Zhuang WY,
Haelewaters D, Park D, Carl S, López-Giráldez F, Wang Z, Townsend JP (2019) A mul-
tigene phylogeny toward a new phylogenetic classication of Leotiomycetes. IMA
Fungus 10(1): 1. https://doi.org/10.1186/s43008-019-0002-x
Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A,
Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious
basic: An integrated and extendable desktop software platform for the organization
and analysis of sequence data. Bioinformatics (Oxford, England) 28(12): 1647–1649.
https://doi.org/10.1093/bioinformatics/bts199
Kress WJ, Erickson DL, Jones FA, Swenson NG, Perez R, Sanjur O, Bermingham E (2009)
Plant DNA barcodes and a community phylogeny of a tropical forest dynamics plot in

211
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Panama. Proceedings of the National Academy of Sciences of the United States of
America 106(44): 8621–18626. https://doi.org/10.1073/pnas.0909820106
Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular evolutionary genetics analysis
version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7): 1870–1874.
https://doi.org/10.1093/molbev/msw054
Levin RA, Wagner WL, Hoch PC, Nepokroeff M, Pires JC, Zimmer EA, Sytsma KJ (2003)
Family-level relationships of Onagraceae based on chloroplast rbcL and ndhF data.
American Journal of Botany 90(1): 107–115. https://doi.org/10.3732/ajb.90.1.107
Lutzoni F, Kauff F, Cox CJ, McLaughlin D, Celio G, Dentinger B, Padamsee M, Hibbett
D, James TY, Baloch E, Grube M, Reeb V, Hofstetter V, Schoch C, Arnold AE, Miad-
likowska J, Spatafora J, Johnson D, Hambleton S, Crockett M, Shoemaker R, Sung
GH, Lücking R, Lumbsch T, O’Donnell K, Binder M, Diederich P, Ertz D, Gueidan C,
Hansen K, Harris RC, Hosaka K, Lim YW, Matheny B, Nishida H, Pster D, Rogers J,
Rossman A, Schmitt I, Sipman H, Stone J, Sugiyama J, Yahr R, Vilgalys R (2004) As-
sembling the fungal tree of life: Progress, classication, and evolution of subcellu-
lar traits. American Journal of Botany 91(10): 1446–1480. https://doi.org/10.3732/
ajb.91.10.1446
McTaggart AR, Shivas RG, Braun U (2007) Annellosympodia orbiculata gen. et sp. nov.
and Scolecostigmina agellariae sp. nov. from Australia. Australasian Plant Patholo-
gy 36(6): 573–579. https://doi.org/10.1071/AP07061
Mel’nik VA, Braun U (2013) Atractilina alinae sp. nov. and Neosporidesmium vietnamense
sp. nov. – two synnematous hyphomycetes from Vietnam. Mycobiota 3: 1–9. https://
doi.org/10.12664/mycobiota.2013.03.01
Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES science gateway for infer-
ence of large phylogenetic trees. 2010 Gateway Computing Environments Workshop
(GCE). IEEE, New Orleans, 8. https://doi.org/10.1109/GCE.2010.5676129
Morelet M (1971) De aliquibus in mycologia novitatibus (5e note). Bulletin de la Société
de Sciences Naturelles et d ’Archéologie de Toulon et du Var 195: 7–8. https://doi.
org/10.2113/gssgfbull.S7-XIII.1-2.195
Nakamura T, Yamada KD, Tomii K, Katoh K (2018) Parallelization of MAFFT for large-
scale multiple sequence alignments. Bioinformatics (Oxford, England) 34(14): 2490–
2492. https://doi.org/10.1093/bioinformatics/bty121
Ordynets A, Keßler S, Langer E (2021) Geometric morphometric analysis of spore
shapes improves identication of fungi. PLOS ONE 16(8): e0250477. https://doi.
org/10.1371/journal.pone.0250477
Pem D, Jeewon R, Chethana KWT, Hongsanan S, Doilom M, Suwannarach N, Hyde KD
(2021) Species concepts of Dothideomycetes: Classication, phylogenetic inconsis-
tencies and taxonomic standardization. Fungal Diversity 109(1): 283–319. https://
doi.org/10.1007/s13225-021-00485-7
Piepenbring M, Hofmann TA, Kirschner R, Mangelsdorff R, Perdomo O, Rodríguez Jus-
tavino D, Trampe T (2011) Diversity patterns of Neotropical plant parasitic microfungi.
Ecotropica (Bonn) 17: 27–40.
Promputtha I, Jeewon R, Lumyong S, McKenzie EHC, Hyde KD (2005) Ribosomal DNA
ngerprinting in the identication of non-sporulating endophytes from Magnolia liliif-
era (Magnoliaceae). Fungal Diversity 20: 167–186.
Promputtha I, Lumyong S, Vijaykrishna D, McKenzie EHC, Hyde KD, Jeewon R (2007)
A phylogenetic evaluation of whether endophytes become saprotrophs at host se-
nescence. Microbial Ecology 53(4): 579–590. https://doi.org/10.1007/s00248-006-
9117-x

212
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Raja HA, Miller AN, Pearce CJ, Oberlies NH (2017) Fungal Identication Using Molecular
Tools: A Primer for the Natural Products Research Community. Journal of Natural
Products 24 80(3): 756–770. https://doi.org/10.1021/acs.jnatprod.6b01085
Rodríguez Justavino D, Kirschner R, Piepenbring M (2015) New species and new records
of Meliolaceae from Panama. Fungal Diversity 70(1): 73–84. https://doi.org/10.1007/
s13225-014-0292-7
Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L,
Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Ecient bayesian phylogenetic
inference and model choice across a large model space. Systematic Biology 61(3):
539–542. https://doi.org/10.1093/sysbio/sys029
Schoch CL, Crous PW, Groenewald JZ, Boehm EWA, Burgess TI, de Gruyter J, de Hoog
GS, Dixon LJ, Grube M, Gueidan C, Harada Y, Hatakeyama S, Hirayama K, Hosoya T,
Huhndorf SM, Hyde KD, Jones EBG, Kohlmeyer J, Kruys Å, Li YM, Lücking R, Lumbsch
HT, Marvanová L, Mbatchou JS, McVay AH, Miller AN, Mugambi GK, Muggia L, Nelsen
MP, Nelson P, Owensby CA, Phillips AJL, Phongpaichit S, Pointing SB, Pujade-Renaud
V, Raja HA, Plata E Rivas, Robbertse B, Ruibal C, Sakayaroj J, Sano T, Selbmann L,
Shearer CA, Shirouzu T, Slippers B, Suetrong S, Tanaka K, Volkmann-Kohlmeyer B,
Wingeld MJ, Wood AR, Woudenberg JHC, Yonezawa H, Zhang Y, Spatafora JW
(2009) A class-wide phylogenetic assessment of Dothideomycetes. Studies in My-
cology 64: 1–15. https://doi.org/10.3114/sim.2009.64.01
Seifert KA, Hughes SJ (2000) Spiropes dictyosporus, a new synnematous fungus associ-
ated with sooty moulds. New Zealand Journal of Botany 38(3): 489–492. https://doi.
org/10.1080/0028825X.2000.9512698
Stamatakis A (2014) RAxML version 8: A tool for phylogenetic analysis and post-analy-
sis of large phylogenies. Bioinformatics (Oxford, England) 30(9): 1312–1313. https://
doi.org/10.1093/bioinformatics/btu033
Stevens FL (1918) Some meliolicolous parasites and commensals from Porto Rico. Bo-
tanical Gazette (Chicago, Ill.) 65(3): 227–249. https://doi.org/10.1086/332230
Talavera G, Castresana J (2007) Improvement of phylogenies after removing divergent
and ambiguously aligned blocks from protein sequence alignments. Systematic Biol-
ogy 56(4): 564–577. https://doi.org/10.1080/10635150701472164
Thambugala KM, Wanasinghe DN, Phillips AJL, Camporesi E, Bulgakov TS, Phukham-
sakda C, Ariyawansa HA, Goonasekara ID, Phookamsak R, Dissanayake A, Tennakoon
DS, Tibpromma S, Chen YY, Liu ZY, Hyde KD (2017) Mycosphere notes 1–50: Grass
(Poaceae) inhabiting Dothideomycetes. Mycosphere : Journal of Fungal Biology 8(4):
697–796. https://doi.org/10.5943/mycosphere/8/4/13
Valenzuela-Lopez N, Magaña-Dueñas V, Cano-Lira JF, Wiederhold N, Guarro J, Stchigel
AM (2019) Two new species of Gloniopsis (Hysteriales, Ascomycota) from clinical
specimens: Morphological and molecular characterization. Mycoses 62(12): 1164–
1173. https://doi.org/10.1111/myc.13006
Vilgalys R, Hester M (1990) Rapid genetic identication and mapping of enzymatically
amplied ribosomal DNA from several Cryptococcus species. Journal of Bacteriology
172(8): 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
Von Arx JA, Müller E (1975) A re-evaluation of the bitunicate ascomycetes with keys to
families and genera. Studies in Mycology 9: 1–159.
Wagner T, Ryvarden L (2002) Phylogeny and taxonomy of the genus Phylloporia
(Hymenochaetales). Mycological Progress 1(1): 105–116. https://doi.org/10.1007/
s11557-006-0009-8

213
MycoKeys 103: 167–213 (2024), DOI: 10.3897/mycokeys.103.115799
Miguel A. Bermúdez-Cova et al.: Species of Atractilina and Spiropes hyperparasitic on Meliolales
Wanasinghe DN, Phukhamsakda C, Hyde KD, Jeewon R, Lee HB, Jones EBG, Tibpromma
S, Tennakoon DS, Dissanayake AJ, Jayasiri SC, Gafforov Y, Camporesi E, Bulgakov
TS, Ekanayake AH, Perera RH, Samarakoon MC, Goonasekara ID, Mapook A, Li W-J,
Senanayake IC, Li J, Norphanphoun C, Doilom M, Bahkali AH, Xu J, Mortimer PE, Tibell
L, Tibell S, Karunarathna SC (2018) Fungal diversity notes 709–839: Taxonomic and
phylogenetic contributions to fungal taxa with an emphasis on fungi on Rosaceae.
Fungal Diversity 89(1): 1–236. https://doi.org/10.1007/s13225-018-0395-7
Wang Z, Johnston PR, Takamatsu S, Spatafora JW, Hibbett DS (2006a) Towards a phy-
logenetic classication of Leotiomycetes based on rDNA data. Mycologia 98(6):
1065–1075. https://doi.org/10.1080/15572536.2006.11832634
Wang Z, Binder M, Schoch CL, Johnston PR, Spatafora JW, Hibbett DS (2006b) Evolution
of helotialean fungi (Leotiomycetes, Pezizomycotina): A nuclear rDNA phylogeny.
Molecular Phylogenetics and Evolution 41(2): 295–312. https://doi.org/10.1016/j.
ympev.2006.05.031
White TJ, Brun T, Lee S, Taylor J (1990) Amplication and direct sequencing of fungal
ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White
TJ (Eds) PCR Protocols: a guide to methods and applications, Academic Press, San
Diego. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
Zeng XY, Zhao JJ, Hongsanan S, Chomnunti P, Boonmee S, Wen TC (2017) A check-
list for identifying Meliolales species. Mycosphere: Journal of Fungal Biology 8(1):
218–359. https://doi.org/10.5943/mycosphere/8/1/16
Zhang K, Guo W, Sosa D, Magdama F, Serrano L, Malosso E, Li D-W, Castañeda-Ruiz RF
(2020) Phylogeny and morphology of Ellismarsporium parvum and the new combina-
tion E. varium. Mycotaxon 135(2): 443–452. https://doi.org/10.5248/135.443
Zhao G, Wu Y, Li N (1996) Six fungi species of hyperparasite on Meliolaceae. Journal of
Beijing Forestry University 18(1): 99–103.
Supplementary material 1
Alignments and tree generated during the analysis of the DNA sequences
of Atractilina parasitica, Malacaria meliolicola and other members of the
Dothideomycetes
Authors: Miguel A. Bermúdez-Cova, Tina A. Hofmann, Nourou S. Yorou, Meike Piepenbring
Data type: docx
Explanation note: Alignment is shown in NEXUS format. The tree is shown in Newick format.
Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.
Link: https://doi.org/10.3897/mycokeys.103.115799.suppl1