A peer-reviewed open-access journal
MycoKeys 44: 81–122 (2018)
Melanospora (Sordariomycetes, Ascomycota) and its relatives
doi: 10.3897/mycokeys.44.29742
RESEARCH ARTICLE
http://mycokeys.pensoft.net
81
MycoKeys
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Melanospora (Sordariomycetes, Ascomycota)
and its relatives
Yasmina Marin-Felix1,2, Josep Guarro1, José F. Cano-Lira1, Dania García1,
Andrew N. Miller3, Alberto M. Stchigel1
1 Mycology Unit, Medical School and IISPV, Universitat Rovira i Virgili, C/ Sant Llorenç 21, 43201 Reus,
Tarragona, Spain 2 Westerdijk Fungal Biodiversity Institute, P.O. Box 85167, 3508 CT Utrecht, Netherlands
3 Illinois Natural History Survey, University of Illinois, 1816 S. Oak St., Champaign, Illinois, USA 61820
Corresponding author: Yasmina Marin-Felix (y.marin@westerdijkinstitute.nl)
Academic editor: T. Lumbsch | Received 13 September 2018 | Accepted 18 November 2018 | Published 18 December 2018
Citation: Marin-Felix Y, Guarro J, Cano-Lira JF, García D, Miller AN, Stchigel AM (2018) Melanospora
(Sordariomycetes, Ascomycota) and its relatives. MycoKeys 44: 81–122. https://doi.org/10.3897/mycokeys.44.29742
Abstract
The order Melanosporales comprises a large group of ascomycetes, most of them mycoparasites,
characterized by the production of usually ostiolate, translucent ascomata, unitunicate asci, and
unicellular, pigmented ascospores with germ pores or germ slits. The most studied taxa are Melanospora
and Sphaerodes, but the boundaries with other morphologically closely related genera are not well resolved.
In this study, the taxonomy of Melanospora and related taxa have been re-evaluated based on the analysis
of nuclear rDNA, actin and elongation factor genes sequences of fresh isolates and numerous type and
reference strains. The genus Melanospora has been restricted to species with ostiolate ascoma whose
neck is composed of intermixed hyphae, and with a phialidic asexual morph. Microthecium has been reestablished for species of Melanospora and Sphaerodes without a typical ascomatal neck or, if present, being
short and composed of angular cells similar to those of the ascomatal wall, and usually producing bulbils.
Three new genera have been proposed: Dactylidispora, possessing ascospores with a raised rim surrounding
both terminal germ pores; Echinusitheca, with densely setose, dark ascomata; and Pseudomicrothecium,
characterized by ascospores with indistinct germ pores. Dichotomous keys to identify the accepted genera
of the Melanosporales, and keys to discriminate among the species of Melanospora and Microthecium, as
well as a brief description of the accepted species of both genera, are also provided.
Keywords
Ceratostomataceae, Dactylidispora, Echinusitheca, Melanosporales, Microthecium, Pseudomicrothecium,
soil, Sphaerodes, 4 new taxa
Copyright Yasmina Marin-Felix et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC
BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Introduction
The family Ceratostomataceae (Winter 1887) includes nearly 100 species, often
mycoparasitic and characterized by ostiolate and rostrate, or less frequently non-ostiolate,
translucent ascomata, unitunicate and evanescent asci, brown or exceptionally hyaline,
unicellular ascospores with a germ pore at each end, or less frequently with only one
germ pore or a germ slit, and phialidic asexual morphs or bulbils. Currently, that family
is included in the Melanosporales (Chaudhary et al. 2006, Zhang et al. 2006, Hibbett
et al. 2007, Li et al. 2016, Schultes et al. 2017), although historically it had been placed
in Aspergillales (Gaümann 1964), Hypocreales (Alexopoulos 1962, Spatafora and
Blackwell 1994a, Rehner and Samuels 1995, Jones and Blackwell 1998, Zhang and
Blackwell 2002) and Sphaeriales (Bessey 1950, Dennis 1968). This family comprises 11
sexually reproducing genera, i.e. Arxiomyces, Melanospora, Persiciospora, Pteridiosperma,
Pustulipora, Rhytidospora, Scopinella, Setiferotheca, Sphaerodes, Syspastospora and Vittatispora.
Melanospora, the largest genus of this family (more than 50 species), was established by
Corda (1837) to accommodate Ceratostoma chionea and two new species, Melanospora
zamiae and Melanospora leucotricha, with the former chosen later as the type species
(Kowalski 1965). Melanospora is characterized by usually ostiolate ascomata with a long
neck and a translucent, pale yellow to reddish brown ascomatal wall, and mostly smoothwalled, brown, ellipsoidal to citriform, rarely discoid or fusoid ascospores, with a depressed
germ pore at each end, occasionally surrounded by a raised rim (Guarro et al. 2012).
Related genera are Microthecium and Sphaerodes. The former was erected by Corda (1842)
to distinguish Mi. zobelii from Melanospora spp. by the presence of non-ostiolate, usually
immersed ascomata; and Sphaerodes was introduced by Clements (1909) to separate
Melanospora episphaeria from Melanospora spp. by its reticulate ascospores. However,
the generic boundaries between Melanospora and its relatives remained obscure. Doguet
(1955) carried out a revision of Melanospora, synonymizing several species and transferring
additional species from other genera, mostly from Sphaeroderma, which had been proposed
by Fuckel (1877) and distinguished from Melanospora by the absence of an ascomatal neck.
Doguet (1955) considered the production of a neck as a non-stable taxonomic character
influenced by the nature of the substrate where the fungus grows, and segregated the
genus in several sections on the basis of the morphology of the ascomata (presence or
absence of neck, and its size when present) and ascospores (shape and ornamentation).
The most comprehensive revision of Melanospora and related genera was carried out by
Cannon and Hawksworth (1982), based mainly on the structure of the ascospore wall
under SEM, resulting in the transfer of species of Microthecium to Melanospora and to
Sphaerodes. However, recent molecular studies demonstrated that these two latter genera
are polyphyletic (Zhang and Blackwell 2002, Fan et al. 2012, Li et al. 2016, Schultes et
al. 2017). Other genera included in the family are: Arxiomyces, which produces ovoid to
ellipsoidal ascospores with a rounded apex and a truncate base with a large sunken germ pore
(Cannon and Hawksworth 1982, 1983); Persiciospora, characterized by ascospores with a
pitted wall and a faint reticulation (Cannon and Hawksworth 1982); Pteridiosperma, with
ascospores ornamented with longitudinal wing-like appendages (Krug and Jeng 1979);
Melanospora (Sordariomycetes, Ascomycota) and its relatives
83
Pustulipora, distinguished by its ascospores with a germ pore at each end surrounded by a
blistered, rarely cushion-like structure showing an irregular pustulate appearance (Cannon
1982); Rhytidospora, characterized by non-ostiolate ascomata with a cephalothecoid
ascomatal wall (Krug and Jeng 1979); Scopinella, producing brown, cuboid-ellipsoidal
ascospores with two prominent longitudinal germ slits (Cannon and Hawksworth 1982);
Setiferotheca, which produces ascospores similar to those of Arxiomyces and ascomata with
a crown of dark brown setae surrounding the ostiole (Matsushima 1995); Syspastospora,
possessing ascomata with a long neck composed of parallel arranged hyphae and
cylindrical ascospores with a large terminal slightly sunken germ pore at each end (Cannon
and Hawksworth 1982); and Vittatispora, which produces ascomata similar to those of
Syspastospora and citriform ascospores with a longitudinal, thick, hyaline ridge (Chaudhary
et al. 2006). Practically all taxonomic studies on these fungi have been based exclusively on
the morphological characterization of the reproductive structures of preserved fungarium
specimens, since unfortunately due to their mycoparasitism, many of these fungi do not
grow in pure culture or do not produce ascomata in absence of their hosts. On the other
hand, obtaining reliable nucleotide sequences from members of the Melanosporales is also
difficult because of the usually large amount of DNA of their hosts. Based on the study
of several freshly-isolated soil-borne fungi and of reference and type strains obtained from
various culture collections, we have re-examined the phylogenetic relationships of the most
relevant genera of the Ceratostomataceae. Consequently, the genus Melanospora has been
redefined, Microthecium has been re-established, and three new genera have been proposed.
Materials and methods
Fungal isolates
The strains included in this study are listed in Table 1. Fresh isolates were obtained
from samples following previously described procedures for the activation of dormant
ascospores in soil using acetic acid and phenol solutions (Stchigel et al. 2001, García et
al. 2003). Ascomata were transferred to 55 mm diam. Petri dishes containing oatmeal
agar (OA; oatmeal flakes, 30 g; agar-agar, 20 g; distilled water, 1 L) using a sterile
needle, which were then incubated at 15, 25 and 35 °C.
Morphological study
For cultural characterization, isolates were grown for up to 30 d on OA, potato carrot
agar (PCA; grated potatoes, 20 g; grated carrot, 20 g; agar-agar, 20 g; L-chloramphenicol,
100 mg; distilled water, 1 L), and potato dextrose agar (PDA; Pronadisa, Madrid,
Spain) at 5, 10, 15, 20, 25, 30, 35 and 40 °C. Color notations in parentheses are from
Kornerup and Wanscher (1984). Vegetative and reproductive structures were examined
under an Olympus BH-1 brightfield microscope by direct mounting in lactic acid
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Table 1. Isolates and reference strains of members of Melanosporales included in the combined
phylogenetic study.
Taxa
Dactylidispora
ellipsospora
Dactylidispora
singaporensis
Echinusitheca
citrispora
Strain
NBRC 31376T
NBRC 30865T
CBS 137837T
= FMR 12767T
Source
LSU
Forest soil, Papua New Guinea, KP981451
Buin, Bougainville Island
Soil, Singapore
KP981452
Nectria cinnabarina
CBS 127383
Melanospora
damnosa
Melanospora
kurssanoviana
Melanospora
verrucispora
Melanospora zamiae
Microthecium
ciliatum
Microthecium
compressum
Microthecium
fayodii
CBS 113681
Forest soil, USA, North
Carolina, Great Smoky
Mountains National
Park, Cataloochee Creek
Campground
Austria, Niederösterreich,
Litschau
Soil, France, Pont d’Espagne
NBRC 8098
Unknown
Microthecium
fimbriatum
Microthecium
fimicola
Microthecium
fusisporum
Microthecium
japonicum
Microthecium
levitum
Microthecium
quadrangulatum
Microthecium
retisporum
Microthecium
sepedonioides
Microthecium sp.
Microthecium sp.
Microthecium
tenuissimum
NBRC 31375T
NBRC 7902
NBRC 9829
KP981453
GenBank accession number
ITS
act
tef1
03137601* KP981545 KP981579
03086502* KP981546 KP981580
KP981477 KP981547 KP981581
HM534894 HM534894
–
HM534873
KP981454
KP981478 KP981543 KP981582
KP981455
KP981479 KP981548 KP981583
Forest soil, Papua New Guinea, KP981456
Kebil, Chimb Dist.
Unknown
KP981457
Soil, unknown
KP981458
KP981480 KP981549 KP981584
00790201* KP981544 KP981585
KP981481 KP981524 KP981586
NBRC 8627
Unkown
KP981459
00862701* KP981525 KP981587
FMR 12363
Soil, Tennessee, Great Smoky
Mountains National Park,
Cosby Creek trail
Unknown
KP981460
KP981482 KP981526 KP981588
KP981461
KP981483 KP981527 KP981589
KP981462
KP981463
KP981464
KP981465
KP981484
KP981485
KP981486
KP981487
KP981466
00880601* KP981532 KP981594
KP981467
KP981488 KP981533 KP981595
KP981468
KP981489 KP981534 KP981596
KP981469
KP981470
KP981490 KP981535 KP981597
KP981491 KP981536 KP981598
KP981471
KP981492 KP981537 KP981599
KP981472
00836601* KP981538 KP981600
KP981473
KP981493 KP981539 KP981601
KP981474
KP981494 KP981540 KP981602
KP981475
KP981495 KP981541 KP981603
KY628706
KY628705
NBRC 8523
NBRC 8354
FMR 5483
FMR 12370
FMR 13418
NBRC 8806
FMR 12371
FMR 6218
= CBS 966.97
FMR 10098
FMR 13884
CBS 112763T
NBRC 8366
FMR 11933
FMR 6725
= CBS 102190
FMR 7183
= CBS 108937
CBS 112764T
Unknown
Soil, Australia, Moara
Soil, Spain, Gran Canaria
Soil, Spain, Aragon, Los Valles
Occidentales
Unknown
Soil, Spain, Gran Canaria, Pico
de Osorio
Soil, Nepal, Bhadgaon
Soil, Nigeria, Enugu, Nsukka
Soil, Spain, Catalonia, Vall
Fosca
Soil, Spain, Asturias, Muniellos
Biological Absolute Reserve
Soil, Japan
Forest soil, Spain, Aragón, valle
de Ordesa
Desert soil, Egypt, Sinai
Forest soil, New South Wales,
Sydney, Blue Mountains
Soil, Spain, Murcia, Sierra
de Espuña, Umbria de Peña
Apartada
KP981528
KP981529
KP981530
KP981531
–
KP981590
KP981591
KP981592
KP981593
–
Melanospora (Sordariomycetes, Ascomycota) and its relatives
Taxa
Strain
Microthecium
NBRC 9442
zobelii
Pseudallescheria
CBS 106.53T
fusoidea
Pseudomicrothecium BJTC FAN1001T
subterraneum
Vittatispora coorgii
BICC 7817T
Source
Decaying carpophore of
Coriolus flabelliformis
Soil, Panama, Guipo
From Tuber indicum, China,
Yunnan
Soil, India, Western Ghats,
Coorg District, Kakkabe
LSU
KP981476
85
GenBank accession number
ITS
act
tef1
00944201* KP981542 KP981604
EF151316
AY878941
–
–
JN247804
–
–
–
DQ017375
–
–
–
BICC: Biocon culture collection, Bangalore, India; BJTC: Capital Normal University, Beijing, China; CBS: Westerdijk Fungal
Biodiversity Institute, Utrecht, the Netherlands; FMR: Facultat de Medicina, Reus, Spain; NBRC: Biological Resource Center,
Chiba, Japan. T indicates type strains. * sequences retrieved from NBRC database.
and water of the ascomata and/or microcultures grown on OA and PDA. Pictures
were obtained with a Zeiss Axio Imager M1 brightfield microscope. The samples for
scanning electron microscopy (SEM) were processed according to Figueras and Guarro
(1988), and SEM micrographs were taken at 15 keV with a Jeol JSM 840 microscope.
Molecular study
The DNA of the fungal isolates (Table 1) was extracted and purified directly from the
colonies according to the Fast DNA Kit protocol (MP Biomedicals, Solon, Ohio). The
amplification of the small subunit (SSU), the D1−D3 domains of the large subunit
(LSU) and the internal transcribed spacer region (ITS) of the nuclear rDNA, and the
fragments of actin (act) and translation elongation factor 1-α (tef1) genes were performed
according to White et al. (1990) (SSU), Vilgalys and Hester (1990) (LSU), Cano et al.
(2004) (ITS), Voigt and Wöstermeyer (2000) (act) and Houbraken et al. (2007) (tef1).
A BigDye Terminator 3.1 cycle sequencing kit (Applied Biosystems Inc., Foster City,
California) was used to sequence both strands with a combination of the same primers
used in the amplification. PCR products were purified and sequenced at Macrogen Europe
(Amsterdam, The Netherlands) with a 3730XL DNA analyzer (Applied Biosystems),
and the consensus sequences were obtained using SeqMan (version 7.0.0; DNASTAR,
Madison, WI, USA). A phylogenetic study based on the analysis of SSU sequences of
the isolates and type and reference strains of the Melanosporales and of some members
of the Chaetosphaeriales, Coniochaetales, Coronophorales, Hypocreales, Microascales,
Sordariales and Xylariales, using Thelebolus ellipsoideus (Thelebolales) as outgroup, was
performed to confirm the taxonomic placement of our isolates. A subsequent study, carried
out to infer the phylogenetic relationships among members of the Melanosporales, was
based on the analysis of a combined data set including the ITS, LSU, act and tef1 sequences
of our isolates and of type and reference strains of a large number of the Melanosporales,
including Nectria cinnabarina and Pseudallescheria fusoidea as outgroups. The MaximumLikelihood (ML) and Bayesian Inference (BI) methods were used in phylogenetic analyses
as described by Hernández-Restrepo et al. (2016). Bootstrap support (BS) ≥70 and posterior
probability values (PP) ≥0.95 were considered significant. The sequences generated in
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this study were deposited in GenBank (Table 1 and Fig. 1) and the alignments used in
the phylogenetic analyses were deposited in TreeBASE (http://purl.org/phylo/treebase/
phylows/study/TB2:S17079). Sequences retrieved from GenBank and NBRC included in
the SSU and combined analyses are shown in Fig. 1 and Table 1, respectively.
Results
The SSU phylogenetic study was based on an alignment of 1023 bp and produced a single
ML tree (Fig. 1) inferred from a RAxML analysis. The members of the Melanosporales
including our isolates were placed in a highly supported main clade (100 % BS / 0.99
PP), and the isolate CBS 137837, whose morphological features did not match any
previously described taxon, occurred as a basal branch clearly separated from the other
Melanosporales, which grouped together with a high support (95 % BS / 0.98 PP) and
separated into three subclades. The first one (96 % BS / - PP), contained most of the isolates
morphologically identified as Melanospora, Persiciospora and Sphaerodes, including the type
and reference strains of Melanospora brevirostris, M. fimbriata, M. fusispora, M. levita, M.
zobelii, Papulaspora sepedonioides, Pteridiosperma ciliatum, Sphaerodes compressa, S. fimicola,
S. retispora, S. quadrangularis and S. tenuissima, without significant genetic variation among
them. The second subclade (80 % BS / - PP) comprised the type strains of Sphaerodes
ellipsospora and Sphaerodes singaporensis and a reference strain of Melanospora kurssanoviana,
which resulted clearly separated from the other two, which grouped with high support (100
% BS / 0.98 PP). In the third subclade (100 % BS / 1 PP) were nested the type species
of Melanospora (M. zamiae), the type strains of Melanospora verrucispora and Sphaerodes
mycoparasitica, and reference strains of Melanospora damnosa and Melanospora tiffanii.
The lengths of the individual alignments used in the combined data set were 802
bp (LSU), 535 bp (ITS), 727 bp (act) and 846 bp (tef1), respectively, and the final total
alignment was 2910 bp. In the ML tree derived from the RAxML analysis of the combined
data set (Fig. 2), the Melanosporales were highly supported (100 % BS / 1 PP) and
subdivided into seven lineages. The first clade (89 % BS / 1 PP; Clade Microthecium)
grouped all our isolates, with the exception of CBS 137837, and type or reference strains
of Melanospora fimbriata, M. fusispora, M. levita, M. zobelii, Papulaspora sepedonioides,
Pteridiosperma ciliatum, Sphaerodes compressa, S. fimicola, S. retispora, S. quadrangularis
and S. tenuissima. All the fungi belonging to this clade have non-ostiolate ascomata, or
when a neck is present, it is short and composed of angular cells similar to those of the
ascomatal wall. Also, bulbils (microsclerotial-like asexual propagules) are present in most
of these species. In spite of the high morphological variability shown by members of this
clade, the loci used in the phylogenetic analysis were not able to separate the species from
each other. The second clade (100% BS / 1 PP; Clade Melanospora) comprised the type
species of Melanospora, M. zamiae, the type strain of M. verrucispora and a reference strain
of M. damnosa. The members of this clade produce ostiolate ascomata with a long neck
composed of hyphae irregularly arranged and ending in a crown of setae. In addition, an
asexual morph is commonly present, which is characterized by solitary, sessile, flask-shaped
phialides producing from rounded to ellipsoidal conidia. The third lineage comprised only
Melanospora (Sordariomycetes, Ascomycota) and its relatives
87
Figure 1. RAxML phylogram obtained from SSU sequences of isolates and type and reference strains
included in the Melanosporales, and strains belonging to the orders Chaetosphaeriales, Coniochaetales,
Coronophorales, Hypocreales, Microascales, Sordariales and Xylariales. Thelebolus ellipsoideus was used
as outgroup. RAxML bootstrap support (BS) values above 70 % and Bayesian posterior probability
scores above 0.95 are shown at the nodes. Type strains of the different species are indicated with T.
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Yasmina Marin-Felix et al. / MycoKeys 44: 81–122 (2018)
Figure 2. RAxML phylogram obtained from the combined ITS, LSU, act and tef1 sequences of our
isolates and type and reference strains of the order Melanosporales. Nectria cinnabarina and Pseudallescheria
fusoidea were used as outgroup. RAxML bootstrap support (BS) values above 70 % and Bayesian posterior
probability scores above 0.95 are shown at the nodes. GenBank accession numbers are indicated in
Table 1. Type strains of the different species are indicated with T.
a reference strain of Melanospora kurssanoviana, which failed to sporulate in pure culture.
The fourth clade (90 % BS / 0.98 PP; Clade Dactylidispora) was composed of the type
strains of Sphaerodes ellipsospora and S. singaporensis, both characterized by ascospores with a
raised rim surrounding the germ pores. Finally, the isolate CBS 137837 and the type strains
of Melanospora subterranea and Vittatispora coorgii formed three independent branches.
The isolate CBS 137837 produces globose, non-ostiolate, densely setose, dark ascomata
and smooth-walled ascospores with a depressed germ pore at each end, while the other
two species of this clade also possess morphological features unique in the Melanosporales,
e.g. ascospores with indistinct germ pores in M. subterranea and with a longitudinal, thick,
hyaline ridge in V. coorgii.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
89
Taxonomy
Key to the accepted genera of the Melanosporales producing sexual morphs
(adapted from Cannon and Hawksworth 1982)
1
–
2
–
3
–
4
–
5
–
6
–
7
–
8
–
9
–
10
–
11
–
Ascospores with two longitudinal germ slits ................................. Scopinella
Ascospores with germ pores ........................................................................2
Ascospores with a broad germ pore and a small basal appendage .................3
Ascospores with a germ pore at each end.....................................................4
Ascomata with a crown of dark brown setae surrounding the ostiole .............
................................................................................................ Setiferotheca
Ascomata without setae...............................................................Arxiomyces
Ascospores oblong or cylindric-fusiform, and germ pores crateriform ...........
...............................................................................................Syspastospora
Ascospores and germ pores otherwise ..........................................................5
Ascomata ostiolate; neck long, composed of hyphae ...................................6
Ascomata non-ostiolate or ostiolate; neck absent or short, conical, composed
of angular cells similar to those of the ascomatal wall ..................................7
Neck composed of irregularly arranged hyphae ........................ Melanospora
Neck composed of parallel arranged hyphae.............................. Vittatispora
Ascospores with indistinct germ pores......................... Pseudomicrothecium
Ascospores with conspicuous germ pores ....................................................8
Germ pores surrounded by hyaline structures .............................................9
Germ pores without such structures..........................................................10
Germ pores with a raised rim ................................................Dactylidispora
Germ pores with a blistered, rarely cushion-like structure ..........Pustulipora
Ascomatal wall cephalothecoid................................................ Rhytidospora
Ascomatal wall not cephalothecoid ...........................................................11
Ascomata dark, densely setose ................................................ Echinusitheca
Ascomata translucent, glabrous or surrounded by hyphae-like hairs ..............
.............................................................................................. Microthecium
Dactylidispora Y. Marín, Stchigel, Guarro & Cano, gen. nov.
MycoBank: MB812079
Type species. Dactylidispora ellipsospora (Takada) Y. Marín, Stchigel, Guarro & Cano.
Holotype and ex-type strain: NBRC 31376.
Description. Ascomata superficial, globose to pyriform, ostiolate or not, yellowishbrown, appearing dark brown when the ascospores are mature, glabrous or setose; necks
cellular, short, conical, with a crown of setae surrounding the ostiole; ascomatal wall
membranaceous, of textura angularis. Paraphyses absent. Asci 8-spored, broadly clavate,
short-stipitate, without apical structures, evanescent. Ascospores one-celled, at first hyaline,
becoming brown to dark brown when mature, fusiform or citriform, umbonate and
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Yasmina Marin-Felix et al. / MycoKeys 44: 81–122 (2018)
truncate at the ends, smooth-walled, with one germ pore at each end; germ pores depressed,
surrounded by a raised rim. Conidiophores reduced to conidiogenous cells. Conidiogenous
cells phialidic, solitary, flask-shaped. Conidia hyaline, subglobose to ovoid, smooth-walled.
Etymology. From Greek δακτυλίδης–, ring, and from Latin –spora, spore, due to the
raised rim that surrounds the germ pores of the ascospores.
Notes. The most distinctive characteristic of Dactylidispora is the production of
smooth-walled ascospores with a germ pore at each end surrounded by a raised rim.
Vittatispora, proposed as a new genus by Chaudhary et al. (2006), also produces a raised
rim surrounding the germ pores. However, both genera can be easily distinguished by the
nature of the ascomatal neck, which is composed of angular cells in Dactylidispora and of
parallel arranged hyphae in Vittatispora; and by the presence of a hyaline ridge running the
entire vertical length of the ascospore between the germ pores in Vittatispora. Moreover,
in our phylogenetic study (Fig. 2), Vittatispora also constituted a lineage independent
from the other members of the Melanosporales. Pustulipora is also morphologically
similar to Dactylidispora being characterized by blistered, rarely cushion-like structures
surrounding the germ pore (Cannon 1982). However, unfortunately, Pustulipora could
not be included into this phylogenetic study since living cultures were not available.
The presence of a raised rim was also described in Melanospora collipora (Stchigel et
al. 1997), which is here transfered to Dactylidispora even though it was not possible to
include this species in the phylogenetic study.
Dactylidispora collipora (Stchigel & Guarro) Y. Marín, Stchigel, Guarro & Cano,
comb. nov.
MycoBank: MB812080
Melanospora collipora Stchigel & Guarro, in Stchigel, Guarro & Figueras, Mycol. Res.
101: 446. 1997. [Basionym]
Notes. This species produces ascomata with a crown of setae around the ostiole,
ellipsoidal ascospores, and bulbils.
Dactylidispora ellipsospora (Takada) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812081
Microthecium ellipsosporum Takada, in Kobayasi et al., Bull. natn. Sci. Mus., Tokyo 16:
527. 1973. [Basionym]
≡ Sphaerodes ellipsospora (Takada) D. García, Stchigel & Guarro, Stud. Mycol. 50: 67.
2004.
Notes. Dactylidispora ellipsospora is characterized by non-ostiolate ascomata, fusiform
ascospores and absence of asexual morph.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
91
Dactylidispora singaporensis (Morinaga, Minoura & Udagawa) Y. Marín, Stchigel,
Guarro & Cano, comb. nov.
MycoBank: MB812082
Melanospora singaporensis Morinaga, Minoura & Udagawa, Trans. Mycol. Soc. Japan
19: 142. 1978. [Basionym]
≡ Sphaerodes singaporensis (Morinaga, Minoura & Udagawa) D. García, Stchigel &
Guarro, Stud. Mycol. 50: 67. 2004.
Notes. Dactylidispora singaporensis is distinguished by its ostiolate ascomata, citriform
ascospores, and phialidic asexual morph.
Echinusitheca Y. Marín, Stchigel, Dania García, Guarro, A.N. Mill. & Cano, gen. nov.
MycoBank: MB812084
Fig. 3
Type species. Echinusitheca citrispora Y. Marín, Stchigel, Dania García, Guarro, A.N.
Mill. & Cano. Holotype and ex-type strain, respectively: CBS H-21596, CBS 137837
= FMR 12767.
Description. Ascomata superficial or immersed, solitary to gregarious, globose,
non-ostiolate, strongly setose, semi-translucent, pale brown to brown, appearing
black when ascospores are mature; setae straight, becoming sinuous toward apex, pale
Figure 3. Echinusitheca citrispora (CBS 137837T). A Ascomata B Asci C, D Ascospores E Depressed
germ pore. Scale bars: 50 μm (A); 10 μm (B); 5 μm (C, D); 1 μm (E).
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brown to brown, non-septate, rarely 1-septate, thick-walled, verrucose to tuberculate,
sometimes branched; ascomatal wall membranaceous, of textura angularis to textura
globulosa. Asci 8-spored, globose to subglobose, non-stipitate, without apical structures.
Ascospores at first hyaline, becoming brown to dark brown when mature, ellipsoidal,
one-celled, smooth-walled, with a depressed germ pore at each end.
Etymology. From Latin echinus–, sea urchin, and from Greek –τείχος, wall, because
of the ascomata resemblance to a sea urchin, due to the abundance of setae.
Notes. This genus is characterized by dark, strongly setose, non-ostiolate ascomata.
Apart from Echinusitheca, the other genera of the Melanosporales characterized by
the production of dark semi-translucent ascomata are Arxiomyces and Scopinella, but
both genera differ from Echinusitheca by the production of long ascomatal necks.
Moreover, Scopinella can be easily distinguished from Echinusitheca by its cuboidellipsoidal ascospores with two prominent longitudinal germ slits, and Arxiomyces by
its ellipsoidal ascospores that are rounded at the apex and truncated at the base, and
with a broad germ pore that bears a mucilaginous and collapsing appendage.
Echinusitheca citrispora Y. Marín, Stchigel, Dania García, Guarro, A.N. Mill. &
Cano, sp. nov.
MycoBank: MB812085
Fig. 3
Type. USA, North Carolina, Great Smoky Mountains National Park, Cataloochee
Creek Campground (35.1375; -83.4915), forest soil, 15 July 2008, A.N. Miller, M.
Calduch and A.M. Stchigel, holotype CBS H-21596, cultures ex-type CBS 137837 =
FMR 12767.
Description. Colonies on PDA attaining a diam. of 70–75 mm after 14 d at
35 °C, cottony and granulose due to the presence of a large number of ascomata, white
with grey to black dots, depressed at the centre and margins fringed; reverse yellowishwhite to pale yellow (4A2 to 4A3) and with olive brown (4F2) dots. Colonies on
OA attaining a diam. of 50–60 mm in 14 d at 35 °C, cottony and granulose due to
the presence of numerous ascomata, margins arachnoid, white to orange white (5A2)
with brownish grey dots (5F2); reverse yellowish-white to golden grey (4A2 to 4C2).
Minimum, maximum, and optimum temperature of growth are 20, 40 and 35 °C,
respectively. Mycelium composed of hyaline to pale yellow, septate, branched, smoothwalled hyphae, 1–3 μm diam. Ascomata non-ostiolate, immersed into the mycelium,
solitary or gregarious, globose, 130–280 μm diam., setose, semi-translucent, pale
brown to brown, appearing black when ascospores are mature; setae straight, becoming
sinuous toward apex, 20–200 μm long, 5–20 μm wide at base, tapering gradually to a
rounded tip of 2–5 μm diam., pale brown to brown, non-septate or rarely 1-septate,
thick-walled, verrucose to tuberculate, sometimes branched at apex; ascomatal wall
membranaceous, 30–40 μm thick, composed of 5–6 layers of flattened cells of 5–30
μm diam. of textura angularis to textura globulosa. Asci 8-spored, globose to subglobose,
Melanospora (Sordariomycetes, Ascomycota) and its relatives
93
20–25 × 15–20 μm, soon evanescent, non-stipitate, without apical structures,
irregularly disposed at the centrum. Ascospores irregularly arranged in the asci, onecelled, at first hyaline, becoming brown to dark brown when mature, smooth- and
thick-walled, ellipsoidal, 20–27 × 10–15 μm, with one germ pore at each end; germ
pores 0.75–2 μm diam., depressed. Asexual morph absent.
Etymology. From Latin citrum-, lemon, and -spora, spore, referring to the lemonshaped ascospores.
Melanospora Corda, Icon. fung. (Prague) 1: 24. 1837, emend.
Fig. 4
Type species. Melanospora zamiae Corda, Icon. fung. (Prague) 1: 24. 1837.
Representative strain: NBRC 7902.
Description. Ascomata superficial to immersed, globose to subglobose, ostiolate,
yellowish-orange or reddish, tomentose or glabrous, usually with a long neck composed
of intermixed hypha, with a crown of rigid, hyaline, septate, smooth- and thick-walled
setae; ascomatal wall membranaceous, translucent, of textura angularis. Periphyses
present. Paraphyses absent. Asci 8-spored, clavate, rounded at apex, without apical
structures, thin-walled, evanescent. Ascospores one-celled, at first hyaline, becoming
brown to dark brown when mature, fusiform, ellipsoidal or citriform, smooth-walled,
reticulate or verrucose, with a terminal apiculate or depressed germ pore at each end.
Asexual morph phialidic, hyaline. Bulbils uncommon.
Notes. This genus is distinguished by translucent ascomata with a neck composed
of intermixed hyphae and with an apical crown of setae, smooth or ornamented
ascospores with an apiculate germ pore at each end, and a phialidic asexual morph.
The neck of Melanospora spp. is morphologically similar to those of Syspastospora
and Vittatispora, which are also composed of hyphae. Syspastospora was introduced
in 1982 by Cannon and Hawksworth to accommodate Melanospora parasitica, with
three additional species described later (S. boninensis, S. cladoniae and S. tropicalis).
This genus differs from Melanospora in the production of cylindrical to barrel-shaped
ascospores with a large, slightly sunken germ pore at both ends (ellipsoidal, citriform
or fusiform, having much smaller, apiculate or depressed germ pores in Melanospora).
Vittatispora can be distinguished from Melanospora by the production of ascospores
with a thick, hyaline, longitudinal ridge and a raised rim surrounding the germ pores.
Moreover, Syspastospora and Vittatispora differs from Melanospora in the structure of
the ascomatal neck, which is composed of hyphae in a parallel arrangement in both
genera (interwoven hyphae in Melanospora).
Melanospora is now restricted to species with ascoma bearing a neck composed
of interwoven hyphae and mostly ending in a crown of setae. This kind of neck
differentiates this genus from Microthecium, which has a neck composed of angular
cells similar to those of the ascomatal wall and possessing a crown of setae surrounding
the ostiole rather than disposed at apex of the neck. The only exception is Melanospora
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Figure 4. Morphological features of the genus Melanospora. Melanospora damnosa (CBS 113681).
A Ascoma B Ascomatal neck D Detail of hyphal neck F Ascospores H Ascospore germinating. Melanospora
zamiae (NBRC 7902) C Ascomatal neck E Detail of ascomatal wall. Melanospora verrucispora (NBRC
31375T) G Ascospores I Phialidic asexual morph. Scale bars: 50 μm (A); 10 μm (B–E, I); 5 μm (F–H).
mycoparasitica that does not have this sort of neck, being short, cellular and without
the crown of setae at the top of this, although this could be due to the fact that it was
described and illustrated at an early stage of ascomal development. In a study on the
development and cytology of Melanospora tiffanii, Kowalski (1965) illustrated early
stages of development with the neck appearing similar to that of M. mycoparasitica.
Long hyphal necks are produced in Melanospora arenaria, Melanospora caprina,
Melanospora chionea, Melanospora langenaria, Melanospora longisetosa and Melanospora
washingtonensis; therefore, these have been kept in the emended genus Melanospora,
although they were not included in the phylogenetic study.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
95
Key to the species of Melanospora
1
–
2
–
3
–
4
–
5
–
6
–
7
–
8
–
9
–
10
–
Ascospores with the surface ornamented .....................................................2
Ascospores smooth-walled ..........................................................................4
Ascospores irregularly verrucose .......................................... M. verrucispora
Ascospores reticulate ...................................................................................3
Ascospores coarsely reticulate .......................................... M. mycoparasitica
Ascospores slightly reticulate ........................................................ M. tiffanii
Ascospores discoid-ellipsoidal .....................................................................5
Ascospores otherwise ..................................................................................7
Asci 4-spored; ascospores 14–19 × 12–14 × 8–9 μm .............. M. longisetosa
Asci 8-spored; ascospores smaller ................................................................6
Neck 250–400 μm long; ascospores 7.5–16 × 6–12 × 4–7 μm ..... M. chionea
Neck 150–200(–260) μm long; ascospores 10.5–12(–13.5) × 9–10.5(–12) ×
7–9 μm ......................................................................... M. washingtonensis
Ascomata usually narrower than 100 μm; ascospores citriform to
rhomboidal ..............................................................................M. damnosa
Ascomata usually broader than 100 μm; ascospores ellipsoidal to citriform ..... 8
Ascomata strongly tomentose; neck 1500–2000 μm long ...........M. caprina
Ascomata weakly or not tomentose; neck shorter than 1500 μm .................9
Neck shorter than 250 μm long ...................................................M. zamiae
Neck longer than 800 μm long .................................................................10
Setae longer than 100 μm ......................................................... M. arenaria
Setae up to 50 μm long ............................................................ M. lagenaria
Melanospora arenaria L. Fisch. & Mont., in Montagne, Annls. Sci. Nat., Bot., sér.
4 5: 337. 1856.
Notes. Melanospora arenaria is characterized by ascomata with a long neck and
ellipsoidal to citriform, smooth-walled ascospores. It is similar to Melanospora
caprina, but differs in having less tomentose ascomata with a shorter neck. Also,
it is similar to M. lagenaria, differing only by the size of the setae at the top of the
ascomatal neck. Molecular data is necessary to confirm that both species correspond
to different species since the size of the setae could be influenced by the culture
media on where these grew.
Melanospora caprina (Fr.) Sacc., Syll. fung. (Abellini) 2: 462. 1883.
Sphaeria caprina Fr., Fl. Danic. 11: tab. 1859, fig. 2. 1825. [Basionym]
≡ Ceratostoma caprinum (Fr.) Fr., Summa veg. Scand., Section Post. (Stockholm):
396. 1849.
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≡ Cerastoma caprinum (Fr.) Quél., Mém. Soc. Émul. Montbéliard, Sér. 2 5: 522. 1875.
= Sphaeria vervecina Desm., Annls Sci. Nat., Bot., sér. 2 17: 13. 1842.
≡ Melanospora vervecina (Desm.) Fuckel, Jb. nassau. Ver. Naturk. 23-24: 126. 1870.
= Melanospora vervecina f. arundinis Sacc., Syll. fung. (Abellini) 2: 461. 1883.
Notes. Melanospora caprina is distinguished from the other species of the genus by its
larger, white, densely tomentose ascomata with a very long neck, and ellipsoidal to
citriform, smooth-walled ascospores with slightly apiculate germ pores.
Melanospora chionea (Fr.) Corda, Icon. fung. (Prague) 1: 24. 1837.
Ceratostoma chioneum Fr., Observ. mycol. (Havniae) 2: 340. 1818. [Basionym]
≡ Sphaeria chionea (Fr.) Fr., Syst. mycol. (Lundae) 2: 446. 1823.
≡ Melanospora chionea var. chionea (Fr.) Corda, Icon. fung. (Prague) 1: 24, tab. 7, fig.
297. 1837.
= Sphaeria biformis var. brachystoma Pers., Syn. meth. fung. (Göttingen) 1: 60. 1801.
≡ Melanospora chionea var. brachystoma (Pers.) Sacc., Syll. fung. (Abellini) 2: 461. 1883.
= Sphaeria leucophaea Fr., Elench. fung. (Greifswald) 2: 92. 1828.
≡ Ceratostoma leucophaeum (Fr.) Fr., Summa veg. Scand., Section Post. (Stockholm):
396. 1849.
≡ Melanospora chionea var. leucophea (Fr.) Sacc., Syll. fung. (Abellini) 2: 461. 1883.
= Melanospora antarctica Speg., Boln Acad. nac. Cienc. Córdoba 11: 233. 1888.
Notes. This species is characterized by white, tomentose ascomata and discoid, smoothwalled ascospores with depressed germ pores.
Melanospora damnosa (Sacc.) Lindau, in Engler & Prantl, Nat. Pflanzenfam., Teil.
I (Leipzig) 1: 353. 1897.
Fig. 4A, B, D, F, H
Sphaeroderma damnosum Sacc., Riv. Patol. veg. 4: 64. 1895. [Basionym]
Notes. Melanospora damnosa is distinguised by the production of ascomata with a short
neck and citriform to rhomboidal, smooth-walled ascospores with a slightly apiculate
germ pore at each end.
Melanospora lagenaria (Pers.) Fuckel, Jb. nassau. Ver. Naturk. 23-24: 126. 1870.
Sphaeria lagenaria Pers., Syn. meth. fung. (Göttingen) 1: 58. 1801. [Basionym]
≡ Ceratostoma lagenaria (Pers.) Fr. [as ‘lagenarium’], Syst. veg., Edn 16: 392. 1827.
≡ Auerswaldia lagenaria (Pers.) Rabenh., Hedwigia 1: 116. 1857.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
97
≡ Cerastoma lagenaria (Pers.) Quél., Mém. Soc. Émul. Montbéliard, Sér. 2 5: 522. 1875.
≡ Phaeostoma lagenaria (Pers.) Munk [as ‘lagenarium’], Dansk bot. Ark. 17: 82. 1957.
= Melanospora lagenaria var. tetraspora Rehm, Hedwigia 30: 259. 1891.
Notes. Melanospora lagenaria is similar to M. caprina, but the former has less tomentose
ascomata with shorter necks ending in a poorly developed crown of setae. This species is
also similar to M. arenaria. For morphological comparison see Notes of the latter species.
Melanospora longisetosa P.F. Cannon & D. Hawksw., J. Linn. Soc., Bot. 84: 130. 1982.
Notes. This species is characterized by the formation of 4-spored asci and discoid,
smooth-walled ascospores.
Melanospora mycoparasitica (Vujan.) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812086
Sphaerodes mycoparasitica Vujan., Mycol. Res. 113: 1173. 2009. [Basionym]
Notes. Melanospora mycoparasitica is distinguished by its fusiform, coarsely reticulate
ascospores.
Melanospora tiffanii Kowalski, Mycologia 57: 279. 1965.
Notes. This species is distinguished by its fusiform, slightly reticulate ascospores.
Melanospora verrucispora Takada, in Kobayasi et al., Bull. natn. Sci. Mus., Tokyo
16: 525. 1973.
Fig. 4G, I
Notes. This species is characterized by irregularly verrucose ascospores.
Melanospora washingtonensis Nitzan, J.D. Rogers & D.A. Johnson, Sydowia 56:
282. 2004.
Notes. This species is similar to M. chionea, but they differ in the length of the neck
[150–200(–266) μm in M. washingtonensis vs. 250–400 μm in M. chionea] and in the
size of the ascospores [10.5–12(–13.5) × 9–10.5(–12) × 7–9 μm in M. washingtonensis
vs. 7.5–16 × 6–12 × 4–7 μm in M. chionea], as well as in the presence of a phialidic
asexual morph in M. washingtonensis.
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Melanospora zamiae Corda., Icon. fung. (Prague) 1: 24. 1837.
Fig. 4C, E
= Melanospora leucotricha Corda, Icon. fung. (Prague) 1: 25. 1837.
= Melanospora coemansii Westend., Bull. Acad. R. Sci. Belg., Cl. Sci., sér. 2 2: 579. 1857.
= Melanospora cirrhata Berk. in Cooke, Grevillea 16: 102. 1888.
= Melanospora globosa Berl., Malpighia 5: 409. 1891.
= Melanospora pampeana Speg., Anal. Mus. nac. Hist. nat. B. Aires 6: 287. 1898.
= Melanospora townei Griffiths, Bull. Torrey bot. Club 26: 434. 1899.
= Melanospora rhizophila Peglion & Sacc., Annls mycol. 11: 16. 1913.
= Melanospora mattiroloana Mirande [as ‘mattiroliana’], Bull. Soc. mycol. Fr. 32:
72. 1916.
= Melanospora schmidtii Sacc., Syll. fung. (Abellini) 24: 650. 1926.
= Melanospora asclepiadis Zerova, J. Inst. Bot. Acad. Sci. Ukraine 12: 155. 1937.
Notes. Melanospora zamiae is characterized by the production of ellipsoidal to citriform,
smooth-walled ascospores with a depressed germ pore at each end. Doguet (1955)
described the presence of bulbils; however, later studies did not mention the presence
of such sort of propagules (Calviello 1973, Cannon and Hawksworth 1982), which
rarely occur in the genus.
Doubtful species
Melanospora aculeata E.C. Hansen, Vidensk. Meddel. Dansk Naturhist. Foren.
Kjøbenhavn 59: 15. 1877.
Notes. Cultures of this species are not available, but it was originally described as
producing small asci (18–21 × 7–8 μm) and ascospores (4–6 × 3–4 μm). This species
produced ostiolate ascomata without a neck, typical of Microthecium; however, such
small ascospores have never been seen in Microthecium.
Melanospora endobiotica Woron., Notul. syst. Inst. cryptog. Horti bot. petropol.
3: 31. 1924.
Notes. Cultures are not available, and no illustrations were included in the protologue.
It was reported as morphologically similar to Melanospora rhizophila [now considered
a synonym of Melanospora zamiae (Doguet 1955)] when it was first described
(Woronichin 1924).
Melanospora (Sordariomycetes, Ascomycota) and its relatives
99
Excluded species
Melanospora arachnophila Fuckel, Jb. nassau. Ver. Naturk. 23–24: 127. 1870.
Notes. This species possesses cylindrical asci and hyaline ascospores, features never seen
in Melanospora. It was previously excluded from Melanospora by Doguet (1955).
Melanospora argadis Czerepan., Nov. sist. Niz. Rast. 3: 177. 1966.
Notes. This species shows morphological features never observed in Melanospora,
e.g. small asci (10–14 × 5–6.5 μm) and olivaceous ascospores (5–5.5 × 3–3.5 μm).
The original description is not detailed enough to ascertain its possible taxonomical
placement.
Melanospora exsola Bat. & H.P. Upadhyay, Atas Inst. Micol. Univ. Recife 2: 331. 1965.
Notes. This species is excluded from Melanospora due to its dark brown, nontranslucent, setose ascomata and its small ascospores (4.5–12 × 4–7 μm), which seem
to indicate a closer relationship with Chaetomium.
Melanospora gigantea (Massee & Crossl.) Massee & Crossl., Fungus Flora of Yorkshire (Leeds): 215. 1905.
Notes. Descriptions of this species and of its basionym, Sphaeroderma gigantea, were
not found.
Melanospora lucifuga (Jungh.) Sacc., Syll. fung. (Abellini) 2: 464. 1883.
Notes. Cultures are not available, and the original description does not mention asci
and ascospores. Therefore, we agree with Doguet (1955) in the exclusion of this fungus
from Melanospora.
Melanospora kurssanoviana (Beliakova) Czerepan., Notul. syst. Sect. cryptog.
Inst. bot. Acad. Sci. U.S.S.R. 15: 84. 1962.
Notes. In our phylogenetic study, M. kurssanoviana was placed in an independent
lineage far from Melanospora. Unfortunately, the only living culture available is sterile.
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We did not find any distinctive morphological feature to differentiate this species from
other members of the Melanosporales in the original description and in the drawing to
introduce it as a new genus.
Melanospora macrospora P. Karst., Hedwigia 30: 299. 1891.
Notes. Doguet (1955) excluded this species due to the production of very large
cylindrical asci (480–500 × 33–36 μm) and ascospores (42–52 × 28–35 μm),
morphological features not observed in any other member of the Melanosporales.
Melanospora octahedrica Pat., Cat. Rais. Pl. Cellul. Tunisie (Paris): 109. 1897.
Notes. This species is transferred to Scopinella due to the morphology of its ascospores,
i.e. octahedral ascospores with two prominent longitudinal germ slits.
Scopinella octahedrica (Pat.) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812087
Basionym. Melanospora octahedrica Pat., Cat. Rais. Pl. Cellul. Tunisie (Paris): 109. 1897.
Melanospora pascuensis Stchigel & Guarro, Mycol. Res. 103: 1305. 1999.
Notes. This species is excluded from Melanospora since its neck is cellular or absent,
instead it is characterized by a dark ring-like structure around the germ pores of the
ascospores (Stchigel et al. 1999). This fungus could represent a new genus since such
structure is unique in the Melanosporales, and these kind of structures resulted in being
phylogenetically informative, as in the case of Dactylidispora, which is distinguished
by its ascospores with a raised rim around the germ pores. The type strain of this
specimen was contaminated with another fungus and it could not be included in the
molecular study.
Melanospora setchellii (Harkn.) Sacc. & P. Syd., Syll. fung. (Abellini) 16: 564. 1902.
Notes. This species is excluded from Melanospora since it produces cylindrical asci with
the ascospores uniseriately disposed, a feature never observed in this genus.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
101
Melanospora vitrea (Corda) Sacc., Syll. fung. (Abellini) 2: 463. 1883.
Sphaeronaema vitreum Corda, Icon. fung. (Prague) 1: 25. 1837. [Basionym]
Notes. Doguet (1955) excluded this species due to its oblong, pale yellow ascospores.
Microthecium Corda, Icon. fung. (Prague) 5: 30, 74. 1842, emend.
Fig. 5
= Sphaerodes Clem., Gen. fung. (Minneapolis): 44, 173. 1909.
= Pteridiosperma J.C. Krug & Jeng, Mycotaxon 10: 44. 1979.
= Persiciospora P.F. Cannon & D. Hawksw., J. Linn. Soc., Bot. 84: 133. 1982.
Type species. Microthecium zobelii Corda, Icon. fung. (Prague) 5: 74. 1842.
Representative strain: NBRC 9442.
Description. Ascomata ostiolate or not, superficial or immersed, globose to
subglobose or pyriform, yellowish-orange, orange-brown or reddish, tomentose or
glabrous; necks short or absent, conical, composed of angular cells similar to those
of the ascomatal wall, usually with a crown of hyaline, septate, smooth- and thickwalled setae around the ostiole; ascomatal wall membranaceous, translucent, of textura
angularis. Periphyses present. Paraphyses absent. Asci 8-spored, clavate, rounded at
apex, without apical structures, thin-walled, evanescent. Ascospores one-celled, at first
hyaline, becoming brown to dark brown when mature, ellipsoidal, fusiform, navicular,
citriform, plataniform or spindle-shaped, smooth, reticulate, pitted or wrinkled, with
a terminal apiculate or depressed germ pore at each end. Asexual morph phialidic,
hyaline. Bulbils usually produced, pale orange to reddish-orange.
Notes. Microthecium has translucent ascomata of textura angularis, cellular
necks short or absent, ascospores smooth-walled or ornamented with a depressed
or apiculate germ pore at each end, often producing bulbils and a phialidic asexual
morph. Dactylidispora, Pustulipora and Pseudomicrothecium produce ascomata similar
to Microthecium. However, the two first genera can be distinguished by the presence
of a raised rim and blistered structure surrounding the germ pores of the ascospores,
respectively, while Pseudomicrothecium differs in the production of 2-spored asci and
ascospores with indistinct germ pores.
The species Mi. africanum, Mi. beatonii, Mi. brevirostratum, Mi. episphaerium, Mi.
foveolatum, Mi. geoporae, Mi. hypomyces, Mi. internum, Mi. lenticulare, Mi. marchicum,
Mi. masonii, Mi. micropertusum, Mi. moureai, Mi. nectrioides, Mi. pegleri and Mi.
perplexum were not included in the phylogenetic study because we could not locate
any specimens since the holotypes or living cultures of most of them are not available.
However, these species were transferred to Microthecium based on their complete and
well-illustrated descriptions.
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Figure 5. Morphological features of the genus Microthecium. Microthecium levitum (FMR 10098). A Nonostiolate ascoma E Asci G Ascospores K Ascospore (SEM). Microthecium fayodii (FMR 12363). B Ostiolate
ascomata F Ascospores O Variable shaped bulbils. Microthecium fimicola (FMR 5483). C Detail of cellular
neck M Ascospores (SEM) P Bulbil. Microthecium quadrangulatum (CBS 112763T). D Crown of setae around
the ostiole L Ascospore SEM. Microthecium retisporum (NBRC 8366). H Ascospores N Asexual morph.
Microthecium japonicum (FMR 12371) I Ascospores J Ascospore SEM. Microthecium sepedonioides (FMR
11933) Q Bulbil. Scale bars: 50 μm (A, B, O); 20 μm (C, D); 10 μm (E–I, P, Q); 5 μm (J, L–N); 2.5 μm (K).
Melanospora (Sordariomycetes, Ascomycota) and its relatives
103
Key to the species of Microthecium
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Sexual morph absent, only producing bulbils ................... Mi. sepedonioides
Sexual morph present..................................................................................2
Ascomata non-ostiolate ...............................................................................3
Ascomata ostiolate ....................................................................................13
Ascospores with an ornamented surface ......................................................4
Ascospores smooth-walled or nearly so........................................................8
Ascospores pitted and with wing-like ridges ......................... Mi. foveolatum
Ascospores coarsely reticulate ......................................................................5
Asci 4-spored ..............................................................................................6
Asci 8-spored ..............................................................................................7
Ascospores (25–)28–34(–40) × 14–18(–20) μm .......................Mi. beatonii
Ascospores 22–28 × 12–15 × 9–11 μm ................................. Mi. perplexum
Ascospores 25–34 × 12–18 μm ........................................ Mi. episphaerium
Ascospores 17–20 × 10–12 × 7–9 μm ...................................Mi. retisporum
Ascomata smaller than 120 μm ..........................................Mi. tenuissimum
Ascomata longer than 120 μm ....................................................................9
Ascospores shorter than 20 μm .................................................................10
Ascospores longer than 20 μm ..................................................................11
Ascospores 15–19 × 11–13 × 8–9 μm, with the narrow faces coarsely reticulate
and the others smooth ........................................................Mi. compressum
Ascospores 10–17 × 8–12 × 9–10 μm, entirely smooth-walled.... Mi. levitum
Ascospores fusiform .............................................................. Mi. hypomyces
Ascospores citriform .................................................................................12
Ascospores 28–30 × 12–13(–15) μm ....................................... Mi. geoporae
Ascospores 18–25 × 8.5–12 × 6–9 μm ........................................ Mi. zobelii
Ascospores with wing-like appendages ......................................................14
Ascospores otherwise ................................................................................15
Ascospores wrinkled, (12–)13–18 × (7–)8–10 μm ....................Mi. ciliatum
Ascospores pitted, (17–)20–22(–24) × 12–14 × 10–12 μm .....Mi. lenticulare
Ascospores ornamentated ..........................................................................16
Ascospores smooth-walled ........................................................................23
Ascospores punctate or punctate-reticulate................................................17
Ascospores reticulate or striate-reticulate ...................................................19
Ascospores punctate, ellipsoidal ............................................ Mi. africanum
Ascospores punctate-reticulate, ellipsoidal-fusiform ..................................18
Ascospores delicately punctate, asexual morph and bulbils present ................
.............................................................................................Mi. japonicum
Ascospores coarsely punctate, asexual morph and bulbils absent...................
................................................................................................ Mi. moreaui
Ascospores striate-reticulate ......................................................................20
Ascospores reticulate .................................................................................21
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20
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21
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22
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23
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25
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26
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27
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29
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30
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Yasmina Marin-Felix et al. / MycoKeys 44: 81–122 (2018)
Ascospores with inconspicuous ridges forming a very coarse reticulum, 18–
22(–28) × 9.5–11(–13) × 8–9 μm..................................Mi. micropertusum
Ascospores without ridges or reticulum, 26–36 × 13–17 μm ......Mi. masonii
Ascospores with 4–6 prominent longitudinal ribs ........ Mi. quadrangulatum
Ascospores without longitudinal ribs ........................................................22
Ascospores spindle-shaped, 19.5–22 × 8.5–11 μm ..................Mi. internum
Ascospores citriform to fusiform, 14–20 × 10–17 μm ............... Mi. fimicola
Crown of setae absent ...........................................................Mi. nectrioides
Crown of setae present around the ostiole .................................................24
Ascospores citriform ............................................................ Mi. marchicum
Ascospores otherwise ................................................................................25
Ascospores ellipsoid to citriform, often somewhat plataniform..................26
Ascospores otherwise ................................................................................28
Bulbils present .............................................................................. Mi. fallax
Bulbils absent ...........................................................................................27
Ascospores 21–34 × 11–17 μm ......................................... Mi. brevirostrum
Ascospores 18–22 × 9–11 μm ............................................. Mi. fimbriatum
Ascospores ellipsoid to fusiform ........................................... Mi. fusisporum
Ascospores ellipsoid to navicular ...............................................................29
Ascospores (9.5–)11–12(–13) × 4–4.5 μm .................................. Mi. pegleri
Ascospores longer than 15 μm ..................................................................30
Ascospores 16–24 × 8–12 μm .....................................................Mi. fayodii
Ascospores 25–30 × 11–15 μm ......................................Mi. brevirostratum
Microthecium africanum (J.C. Krug) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812088
Persiciospora africana J.C. Krug, Mycologia 80: 416. 1988. [Basionym]
Notes. Microthecium africanum is characterized by ostiolate ascomata and punctate,
ellipsoidal ascospores. Two asexual morphs with different conidia have been reported:
(i), 1–4(–5)-celled, globose and smooth-walled at first but becoming cylindrical and
coarsely verrucose later; (ii), 1–2-celled, large, usually cylindrical and smooth-walled
(Krug 1988). However, the type strain was probably not a pure culture because the
SSU and LSU sequences match with different species of Fusarium and the pictures
of the conidia type (i) resemble the chlamydospores produced by several species of
this genus.
Microthecium beatonii D. Hawksw., Trans. Mycol. Soc. Japan 18: 145. 1977.
≡ Sphaerodes beatonii (D. Hawksw.) P.F. Cannon & D. Hawksw., Bot. J. Linn. Soc.
84: 145. 1982.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
105
Notes. This species is characterized by non-ostiolate ascomata, 4-spored asci and very
coarsely reticulate, citriform ascospores. These morphological features are also observed
in Microthecium perplexum, but this species produces ascospores with only a third of
the surface coarsely reticulate while the rest remains smooth-walled. Microthecium
episphaerium and Mi. retisporum differ from Mi. beatonii in the production of 8-spored
asci. Moreover, Mi. retisporum produces a phialidic asexual morph and bulbils, which
are absent in the other mentioned species, and smaller ascospores (17–20 × 10–12 ×
7–9 μm) than in Mi. beatonii [28–34(–40) × 14–18(–20) μm], in Mi. episphaerium
(25–34 × 12–18 μm) and in Mi. perplexum (22–28 × 12–15 × 9–11 μm).
Microthecium brevirostratum (Moreau) Y. Marín, Stchigel, Guarro & Cano,
comb. nov.
Mycobank: MB812089
Melanospora brevirostrata Moreau, Bull. Trimest. Soc. mycol. Fr. 61: 59. 1945. [Basionym]
Notes. Microthecium brevirostratum together with Mi. fayodii and Mi. pegleri produces
ostiolate ascomata, smooth-walled, ellipsoidal to navicular or citriform ascospores and
bulbils. Microthecium brevirostratum is easily distinguished by ascospores with apiculate
germ pores and the presence of a phialidic asexual morph (ascospores show depressed
germ pores and lack an asexual morph in other species). Microthecium fayodii and Mi.
pegleri differ in the size of the ascospores, Mi. pegleri having the smallest ascospores in
Microthecium [(9.5–)11–12(–13) × 4–4.5 μm].
Microthecium brevirostrum (Fuckel) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812090
Ceratostoma brevirostre Fuckel, Bot. Ztg. 19: 250. 1861. [Basionym]
≡ Melanospora brevirostris (Fuckel) Höhn., Sber. Akad. Wiss. Wien, Math.-naturw.
Kl., Abt. 1 123: 94. 1914.
= Ceratostoma helvellae Cooke, Grevillea 1: 175. 1873.
≡ Melanospora helvellae (Cooke) Sacc., Syll. fung. (Abellini) 2: 462. 1883.
= Melanospora sphaerodermoides Grove, J. Bot., Lond. 23: 132. 1885.
= Melanospora sphaerodermoides var. sphaerodermoides Grove, J. Bot., Lond. 23: 132.
1885.
= Thielavia soppittii Crossl., Naturalist, London: 7. 1901.
= Rosellinia aurea McAlpine, Fungus Diseases of stone-fruit trees in Australia: 102.
1902.
≡ Sphaeroderma aureum (McAlpine) Sacc. & D. Sacc., Syll. fung. (Abellini) 17: 781.
1905.
≡ Melanospora aurea (McAlpine) Doguet, Botaniste 39: 124. 1955.
= Melanospora sphaerodermoides var. rubella Pidopl., Mikrobiol. Zh. 9: 61. 1948.
106
Yasmina Marin-Felix et al. / MycoKeys 44: 81–122 (2018)
= Melanospora camelina Faurel & Schotter, Revue Mycol., Paris 30: 144. 1965.
= Melanospora tulasnei Udagawa & Cain, Can. J. Bot. 47: 1932. 1970.
Notes. Microthecium brevirostrum, Mi. fallax and Mi. fimbriatum produce ostiolate ascomata
and ellipsoidal to citriform, often plataniform, smooth-walled ascospores with an apiculate
germ pore at each end. Microthecium fimbriatum is easily distinguished by its smaller (100–
110 μm diam.), reddish ascomata, while Mi. fallax differs in the production of bulbils.
Microthecium ciliatum Udagawa & Takada, Trans. Mycol. Soc. Japan 15: 23. 1974.
≡ Pteridiosperma ciliatum (Udagawa & Y. Takada) J.C. Krug & Jeng, Mycotaxon 10:
45. 1979.
Notes. This species is characterized by non-ostiolate ascomata and ellipsoidal to fusiform
ascospores ornamented with wing-like appendages and wrinkles, and the production of
a phialidic asexual morph and bulbils. Microthecium lenticulare and Mi. foveolatum also
present ascospores with wing-like appendages, but these are pitted and not wrinkled (as
in Mi. ciliatum), and neither species produces bulbils. Microthecium foveolatum and Mi.
ciliatum are characterized by non-ostiolate ascomata and the production of a phialidic
asexual morph, whereas Mi. lenticulare has ostiolate ascomata and lacks an asexual morph.
Microthecium compressum Udagawa & Cain, Can. J. Bot. 47: 1921. 1970.
≡ Sphaerodes compressa (Udagawa & Cain) P.F. Cannon & D. Hawksw., J. Linn. Soc.,
Bot. 84: 145. 1982.
Notes. This species is distinguished by the production of non-ostiolate ascomata and
citriform, bilaterally flattened ascospores, with the narrow faces coarsely reticulate and
the widest faces smooth or nearly so, along with the production of a phialidic asexual
morph.
Microthecium episphaerium (W. Phillips & Plowr.) Höhn., Sber. Akad. Wiss.
Wien, Math.-naturw. Kl., Abt. 1 123: 98. 1914.
Melanospora episphaeria W. Phillips & Plowr., Grevillea 10: 71. 1881. [Basionym]
≡ Sphaeroderma episphaerium (W. Phillips & Plowr.) Sacc., Syll. fung. (Abellini) 2:
460. 1883.
≡ Sphaerodes episphaerium (W. Phillips & Plowr.) Clem. [as ‘episphaericum’], Gen.
fung. (Minneapolis): 1‒227. 1909.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
107
≡ Vittadinula episphaeria (W. Phillips & Plowr.) Clem. & Shear, Gen. fung., Edn 2
(Minneapolis): 281. 1931.
= Sphaeroderma epimyces Höhn., Sitzungsberichte der Kaiserlichen Akademie der Wissenschaften Math.-naturw. Klasse Abt. I 116: 103. 1907.
≡ Melanospora epimyces (Höhn.) Doguet, Botaniste 39: 125. 1955.
Notes. Microthecium episphaerium shows non-ostiolate ascomata and very coarsely
reticulate, citriform ascospores. For morphological comparison see Notes of Mi.
beatonii.
Microthecium fallax (Zukal) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812772
Melanospora fallax Zukal, Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1 98: 547.
1889. [Basionym]
= Melanospora anomala Hotson, Proc. Amer. Acad. Arts & Sci 48.: 257. 1912.
= Melanospora cervicula Hotson, Proc. Amer. Acad. Arts & Sci. 48: 254. 1912.
= Melanospora papillata Hotson, Proc. Amer. Acad. Arts & Sci 48.: 251. 1912.
= Melanospora phaseoli Roll-Hansen, Blyttia 6: 73. 1948.
Notes. This species is characterized by ostiolate ascomata, ellipsoidal to citriform, often
plataniform, smooth-walled ascospores, and production of bulbils. For morphological
comparison see Notes of Mi. brevirostrum.
Microthecium fayodii (Vuill.) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812091
Fig. 5B, F, O
Melanospora fayodii Vuill. [as ‘fayodi’], Bull. Séanc. Soc. Sci. Nancy, Sér. 2 8: 33. 1887.
[Basionym]
Notes. This species is characterized by ostiolate ascomata, ellipsoidal to navicular or
citriform, smooth-walled ascospores, and production of bulbils. For morphological
comparison see Notes of Mi. brevirostratum.
Microthecium fimbriatum (Rostr.) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812092
Sphaeroderma fimbriatum Rostr., Oest. Grönl. Svampe: 25. 1894. [Basionym]
≡ Melanospora fimbriata (Rostr.) Petch, Trans. Br. mycol. Soc. 21: 253. 1938.
108
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Notes. Microthecium fimbriatum produces ostiolate ascomata, and citriform to
plataniform, smooth-walled ascospores with a strongly apiculate and tuberculate germ
pore at each end. Although the ascomata was described as small and reddish in the
protologue, the strain included in this study (NBRC 8523) shows larger (250–380
μm diam.), orange-brown ascomata. Moreover, our isolate produces bulbils. For
morphological comparison see Notes of Mi. brevirostrum.
Microthecium fimicola (E.C. Hansen) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812093
Fig. 5C, M, P
Melanospora fimicola E.C. Hansen, Vidensk. Meddel. Dansk Naturhist. Foren. Kjøbenhavn 59: 15. 1876. [Basionym]
≡ Sphaeroderma fimicola (E.C. Hansen) Sacc., Syll. fung. (Abellini) 2: 460. 1883.
≡ Sphaerodes fimicola (E.C. Hansen) P.F. Cannon & D. Hawksw., J. Linn. Soc., Bot.
84: 146. 1982.
= Melanospora ornata Zukal, Verh. zool.-bot. Ges. Wien 35: 340. 1886.
≡ Sphaerodes ornata (Zukal) Arx, Gen. Fungi Sporul. Cult., Edn 3 (Vaduz): 156. 1981.
= Sphaeroderma hulseboschii Oudem., Contrib. Flora Mycol. d. Pays-Bas 11: 23. 1886.
≡ Melanospora hulseboschii (Oudem.) Doguet, Botaniste 39: 121. 1955.
= Melanospora affine Sacc. & Flageolet, Bull. Soc. Mycol. Fr. 12: 67. 1896.
= Melanospora manginii Vincens [as ‘mangini’], Bull. Soc. Mycol. Fr. 33: 69. 1917.
≡ Sphaerodes manginii (Vincens) Arx, Gen. Fungi Sporul. Cult., Edn 3 (Vaduz): 156. 1981.
Notes. Microthecium fimicola is characterized by ostiolate ascomata and coarsely
reticulate ascospores with strongly apiculate germ pores at both ends. The other
species with ostiolate ascomata and reticulate ascospores are Mi. internum and Mi.
quadrangulatum. The main differences among them are the shape and size of the
ascospores, being citriform in Mi. fimicola, spindle-shaped in Mi. internum and
fusiform in Mi. quadrangulatum. The production of bulbils has only been observed in
our fresh isolates of Mi. fimicola, although this was not previously reported.
Microthecium foveolatum Udagawa & Y. Horie, in Hawksworth & Udagawa,
Trans. Mycol. Soc. Japan 18: 149. 1977.
≡ Pteridiosperma foveolatum (Udagawa & Y. Horie) J.C. Krug & Jeng, Mycotaxon 10:
45. 1979.
Notes. This species is easily distinguished by its non-ostiolate ascomata, ellipsoidal to
fusiform ascospores ornamented with small pores and thick wing-like ridges usually
Melanospora (Sordariomycetes, Ascomycota) and its relatives
109
longitudinal but often oblique, and production of phialidic asexual morph. For
morphological comparison see Notes of Mi. ciliatum.
Microthecium fusisporum (Petch) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812094
Sphaeroderma fusisporum Petch, Naturalist, London: 58. 1936. [Basionym]
≡ Melanospora fusispora (Petch) Doguet, Botaniste 39: 215. 1955.
= Melanospora fusispora var. fusispora (Petch) Doguet, Botaniste 39: 215. 1955.
= Melanospora fusispora var. parvispora Matsush., Matsush. Mycol. Mem. 8: 24. 1995.
Notes. Microthecium fusisporum is related to Mi. nectrioides, both possessing ostiolate
ascomata and smooth-walled ascospores. However, Mi. nectrioides can be distinguished
by the absence of the crown of setae around the ostiole and its citriform ascospores,
being fusiform in Mi. fusisporum.
Microthecium geoporae (W. Oberm.) Höhn., Sber. Akad. Wiss. Wien, Math.naturw. Kl., Abt. 1 123: 98. 1914.
Guttularia geoporae W. Oberm., Mykol. Zentbl. 3: 9. 1913. [Basionym]
Notes. This species produces non-ostiolate ascomata and citriform, smoothwalled ascospores. Other species previously placed in Melanospora characterized by
the production of non-ostiolate ascomata and smooth-walled ascospores are Mi.
hypomyces, Mi. levitum and Mi. zobelii. Microthecium hypomyces is distinguished by its
fusiform ascospores (citriform in the other species), and Mi. levitum by the presence
of bulbils and a phialidic asexual morph. Microthecium geoporae and Mi. zobelii are
distinguished by the size of their ascospores [28–30 × 12–13(–15) μm in Mi. geoporae
and 18–25 × 8.5–12 × 6–9 μm in Mi. zobelii]. Microthecium tenuissimum shows
similar morphological features to these species but its ascospores are finely reticulate
under SEM and its ascomata are smaller (less than 120 μm) than in the other species.
Microthecium hypomyces (Höhn.) Höhn., Sber. Akad. Wiss. Wien, Math.-naturw.
Kl., Abt. 1 123: 50. 1914.
Sphaeroderma hypomyces Höhn., Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1
116: 102. 1907. [Basionym]
≡ Melanospora hypomyces (Höhn.) Doguet, Botaniste 39: 215. 1955.
Notes. This species is characterized by non-ostiolate ascomata and fusiform, smoothwalled ascospores. For morphological comparison see Notes of Mi. geoporae.
110
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Microthecium internum (Tehon & G.L. Stout) Y. Marín, Stchigel, Guarro & Cano,
comb. nov.
MycoBank: MB812095
Melanospora interna Tehon & G.L. Stout, Mycologia 21: 181. 1929. [Basionym]
Notes. This species produces ostiolate ascomata and spindle-shaped ascospores with a
coarse and irregular reticulum. For morphological comparison see Notes of Mi. fimicola.
Microthecium japonicum (Y. Horie, Udagawa & P.F. Cannon) Y. Marín, Stchigel,
Guarro & Cano, comb. nov.
MycoBank: MB812096
Fig. 5I, J
Persiciospora japonica Y. Horie, Udagawa & P.F. Cannon, Mycotaxon 25: 233. 1986.
[Basionym]
Notes. Microthecium japonicum is characterized by ostiolate ascomata and ellipsoidal
to fusiform, punctate-reticulate ascospores, similar to Mi. moureai. However, Mi.
japonicum produces a phialidic asexual morph and bulbils (absent in Mi. moureai) and
delicately reticulate ascospores (coarsely reticulate in Mi. moureai).
Microthecium lenticulare (Udagawa & T. Muroi) Y. Marín, Stchigel, Guarro &
Cano, comb. nov.
MycoBank: MB812097
Pteridiosperma lenticulare Udagawa & T. Muroi [as ‘lenticularis’], Trans. Mycol. Soc.
Japan 22: 20. 1981. [Basionym]
Notes. Microthecium lenticulare produces ostiolate ascomata and pitted-walled ascospores
with wing-like appendages. For morphological comparison see Notes of Mi. ciliatum.
Microthecium levitum Udagawa & Cain, Can. J. Bot. 47: 1917. 1970.
Fig. 5A, E, G, K
≡ Sphaerodes levita (Udagawa & Cain) D. García, Stchigel & Guarro, Stud. Mycol.
50: 67. 2004.
Notes. This species is characterized by non-ostiolate ascomata, citrifrom and
smooth-walled ascospores with umbonate and tuberculate germ pores, presence of
Melanospora (Sordariomycetes, Ascomycota) and its relatives
111
bulbils and phialidic asexual morph. For morphological comparison see Notes of
Mi. geoporae.
Microthecium marchicum (Lindau) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812099
Chaetomium marchicum Lindau, Hedwigia 35: 56. 1896. [Basionym]
≡ Sphaeroderma marchicum (Lindau) Sacc. & P. Syd., Syll. fung. (Abellini) 14:
627. 1899.
Notes. Microthecium marchicum is characterized by its ostiolate ascomata and
citrifrom, smooth-walled ascospores. Its ascospores are similar to those of Mi.
geoporae, Mi. hypomyces, Mi. levitum and Mi. zobelii, but all of them produce nonostiolate ascomata.
Microthecium masonii (Kirschst.) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812100
Ceratostoma masonii Kirschst., Trans. Br. mycol. Soc. 18: 306. 1934. [Basionym]
≡ Persiciospora masonii (Kirschst.) P.F. Cannon & D. Hawksw., J. Linn. Soc., Bot. 84:
135. 1982.
Notes. Microthecium masonii is characterized by ostiolate ascomata and ellipsoidal to
fusiform, faintly striate-reticulate ascospores. The same type of ascospore ornamentation
is also observed in Mi. micropertusum, but this latter species is easily distinguished by
the presence of inconspicuous ridges forming a very coarse reticulum and a phialidic
asexual morph.
Microthecium micropertusum (Y. Horie, Udagawa & P.F. Cannon) Y. Marín, Stchigel,
Guarro & Cano, comb. nov.
MycoBank: MB812101
Sphaerodes micropertusa Y. Horie, Udagawa & P.F. Cannon, Mycotaxon 25: 236. 1986.
[Basionym]
Notes. Microthecium micropertusum is distinguished by its ostiolate ascomata, fusiform
to citriform or nearly rhombic in outline ascospores with inconspicuous ridges forming
a coarse reticulum, and presence of phialidic asexual morph. For morphological
comparison see Notes of Mi. masonii.
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Microthecium moreaui (P.F. Cannon & D. Hawksw.) Y. Marín, Stchigel, Guarro
& Cano, comb. nov.
MycoBank: MB812102
Persiciospora moreaui P.F. Cannon & D. Hawksw., J. Linn. Soc., Bot. 84: 134. 1982.
[Basionym]
Notes. Microthecium moreaui is characterized by its ostiolate ascomata, ellipsoidal and
pitted-walled ascospores, and production of bulbils. For morphological comparison
see Notes of Mi. japonicum.
Microthecium nectrioides (Marchal) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812103
Sphaeroderma nectrioides Marchal, Bull. Soc. R. Bot. Belg. 23: 25. 1884. [Basionym]
≡ Melanospora nectrioides (Marchal) Doguet, Botaniste 39: 121. 1955.
= Melanospora asparagi G. Arnaud, Ann. Serv. Epiph. 2: 273. 1915.
Notes. This species produces ostiolate ascomata and citriform, smooth-walled
ascospores. For morphological comparison see Notes of Mi. fusisporum.
Microthecium pegleri (D. Hawksw. & A. Henrici) Y. Marín, Stchigel, Guarro &
Cano, comb. nov.
MycoBank: MB812104
Melanospora pegleri D. Hawksw. & A. Henrici, Kew Bull. 54: 795. 1999. [Basionym]
Notes. Microthecium pegleri is characterized by ostiolate ascomata, ellipsoidal to
plano-convex, smooth-walled ascospores and presence of bulbils. For morphological
comparison see Notes of Mi. brevirostratum.
Microthecium perplexum D. Hawksw., Trans. Mycol. Soc. Japan 18: 151. 1977.
≡ Sphaerodes perplexa (D. Hawksw.) P.F. Cannon & D. Hawksw., Bot. J. Linn. Soc.
84: 148. 1982.
Notes. This species produces non-ostiolate ascomata, 4-spored asci and citrifrom
ascospores usually with smooth walls, but one third of these are coarsely reticulated.
For morphological comparison see Notes of Mi. beatonii.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
113
Microthecium quadrangulatum (D. García, Stchigel & Guarro) Y. Marín, Stchigel,
Guarro & Cano, comb. nov.
MycoBank: MB812105
Fig. 5D, L
Sphaerodes quadrangularis D. García, Stchigel & Guarro, Stud. Mycol. 50: 64. 2004.
[Basionym]
Notes. Microthecium quadrangulatum is characterized by ostiolate ascomata and
fusiform, reticulate ascospores with strongly apiculate germ pores. For morphological
comparison see Notes of Mi. fimicola.
Microthecium retisporum Udagawa & Cain, Can. J. Bot. 47: 1926. 1970.
Fig. 5H, N
≡ Sphaerodes retispora (Udagawa & Cain) P.F. Cannon & D. Hawksw., J. Linn. Soc.,
Bot. 84: 149. 1982.
= Microthecium retisporum var. inferius Udagawa & Cain [as ‘inferior’], Can. J. Bot.
47: 1928. 1970.
≡ Sphaerodes retispora var. inferior (Udagawa & Cain) P.F. Cannon & D. Hawksw., J.
Linn. Soc., Bot. 84: 149. 1982.
≡ Sphaerodes inferior (Udagawa & Cain) D.W. Li & N.P. Schultes, in Schultes, Murtishi
& Li, Fungal Biology 121: 901. 2017.
= Microthecium retisporum var. retisporum Udagawa & Cain, Can. J. Bot. 47: 1926. 1970.
≡ Sphaerodes retispora var. retispora (Udagawa & Cain) P.F. Cannon & D. Hawksw., J.
Linn. Soc., Bot. 84: 149. 1982.
Notes. This species is characterized by non-ostiolate ascomata, reticulate citriform
ascospores with apiculate germ pores, a phialidic asexual morph and presence of
bulbils. For morphological comparison see Notes of Mi. beatonii.
Microthecium sepedonioides (Preuss) Y. Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812106
Fig. 5Q
Papulaspora sepedonioides Preuss, Linnaea 24: 112. 1851. [Basionym]
Notes. Microthecium sepedonioides only produces bulbils and the sexual morph has
never been observed.
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Yasmina Marin-Felix et al. / MycoKeys 44: 81–122 (2018)
Microthecium tenuissimum (D. García, Stchigel & Guarro) Y. Marín, Stchigel,
Guarro & Cano, comb. nov.
MycoBank: MB812107
Sphaerodes tenuissima D. García, Stchigel & Guarro, Stud. Mycol. 50: 65. 2004.
[Basionym]
Notes. This species is characterized by non-ostiolate ascomata and citriform, ellipsoidal
in lateral view, finely reticulate ascospores with strongly apiculate germ pores. For
morphological comparison see Notes of Mi. geoporae.
Microthecium zobelii Corda, Icon. fung. (Prague) 5: 74. 1842.
≡ Sphaeria zobelii (Corda) Tul. & C. Tul., Fungi hypog.: 186. 1851.
≡ Ceratostoma zobelii (Corda) Berk., Journal of the Royal Horticultural Society 4:
402. 1860.
≡ Melanospora zobelii (Corda) Fuckel, Jb. nassau. Ver. Naturk. 23-24: 127. 1870.
= Melanospora zobelii var. zobelii (Corda) Fuckel, Jb. nassau. Ver. Naturk. 23-24: 127. 1870.
= Melanospora coprophila Zukal, Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1
98: 544. 1889.
= Melanospora marchicum Lindau, Hedwigia 35: 56. 1896.
= Melanospora zobelii var. minor Pidopl., Mikrobiol. Zh. 9(2-3): 60. 1948.
Notes. Microthecium zobelii produces non-ostiolate ascomata, and citriform, smoothwalled ascospores with slightly apiculate germ pores. For morphological comparison
see Notes of Mi. geoporae.
Doubtful species
Microthecium ryvardenianum Aramb. & Gamundí, Agarica 6: 124. 1985.
Notes. This species is considered as doubtful because it presents morphological features
atypical of Microthecium (e.g. allantoid ascospores when immature becoming striate
when mature).
Pseudomicrothecium Y. Marín, Stchigel, Guarro, Cano, gen. nov.
MycoBank: MB812108
Type species. Pseudomicrothecium subterraneum (L. Fan, C.L. Hou, P.F. Cannon &
Yong Li) Y. Marín, Stchigel, Guarro & Cano. Holotype and ex-isotype strain: BJTC
FAN1001, K[M] 172128.
Melanospora (Sordariomycetes, Ascomycota) and its relatives
115
Description. Ascomata non-ostiolate, globose, translucent, pale brown to brown,
appearing dark brown when the ascospores are mature, glabrous or setose; ascomatal
wall membranaceous, of textura angularis. Asci 2-spored, clavate, short-stipitate,
without apical structures, evanescent. Ascospores one-celled, at first hyaline, becoming
dark brown to blackish when mature, ellipsoidal to citriform, umbonate and truncate
at both ends, with a terminal indistinct germ pore at each end. Asexual morph absent.
Etymology. The name refers to the morphological resemblance to Microthecium.
Notes. The new genus Pseudomicrothecium is proposed here to accommodate
Melanospora subterranea because it constitutes a separate lineage in our phylogenetic
study. This genus is characterized by its non-ostiolate ascomata, similar to those of
Microthecium, 2-spored asci and smooth-walled ascospores with an indistinct germ pore
at each end. Asci containing two ascospores have only been observed in some species of
Scopinella (i.e. Scopinella gallicola and S. sphaerophila). However, Scopinella can be easily
distinguished from Pseudomicrothecium by the production of ostiolate ascomata with
long necks and cuboid-ellipsoidal ascospores with two prominent longitudinal germ slits.
Pseudomicrothecium subterraneum (L. Fan, C.L. Hou, P.F. Cannon & Yong Li) Y.
Marín, Stchigel, Guarro & Cano, comb. nov.
MycoBank: MB812109
Basionym. Melanospora subterranea L. Fan, C.L. Hou, P.F. Cannon & Yong Li,
Mycologia 104: 1434. 2012.
Discussion
We have revised the taxonomy of relevant members of the family Ceratostomataceae
based on the analyses of the SSU, LSU, ITS, act and tef1 nucleotide sequences. This
study strongly supported the order Melanosporales proposed by Zhang and Blackwell
in 2007 (Hibbett et al. 2007). The phylogenetic inference showed seven lineages
corresponding to the genera Dactylidispora, Echinusitheca, Melanospora, Microthecium,
Pseudomicrothecium and Vittatispora, and to Melanospora kurssanoviana. Our results
agree with previous studies (Zhang and Blackwell 2002, Fan et al. 2012) which already
suggested and demonstrated that the ornamentation of the ascospores under SEM, a
feature traditionally used to delimit most of the genera in the Melanosporales, is not
useful for estimating phylogenetic relationships among these fungal taxa. Similarly, the
morphology of the ascospores is of weak taxonomic value and a poor predictor for the
generic delimitation of members of the family Sordariaceae, resulting in the synonymy of
two relevant genera, i.e. Gelasinospora and Neurospora (Dettman et al. 2001, García et al.
2004, Nygren et al. 2011). In our study, two of the largest genera of the Melanosporales,
Melanospora and Microthecium, grouped species with both smooth and ornamented
ascospore walls. By contrast, a phylogenetic study of the Lasiosphaeriaceae (Miller
and Huhndorf 2005) revealed that the morphology of the ascomatal wall was more
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phylogenetically informative than that of the ascospores, with several new genera proposed
(i.e. Immersiella) or emended (i.e. Lasiosphaeria, Lasiosphaeris and Schizothecium) (Miller
and Huhndorf 2004, Cai et al. 2005). Here, the erection of the new genus Echinusitheca
is a clear example of the relevance of the ascomatal morphology in the taxonomy of
these fungi, and in fact this taxon together with Arxiomyces and Scopinella are the only
genera in the Melanosporales that show dark semi-transluscent ascomata. In this context,
although Echinusitheca has ascospores similar to those of Melanospora and Microthecium,
this genus constitutes one of the lineages phylogenetically most distant within this order.
Another lineage considerably distant from the other members of the Melanosporales
is constituted by the clade represented only by the species Melanospora kurssanoviana,
suggesting that this fungus could represent a new genus. However, this new taxon is at
this moment not proposed because its colonies, in spite of attempts to induce sporulation,
remain sterile and a detailed morphological study was not possible. The infertility of the
cultures is probably due to the fact that an important part of the members of this fungal
group show a peculiar habitat developing a certain degree of mycoparasitism and requiring
the presence of the host to complete the biologic cycle and develope reproductive structures.
The mycoparasitism of Melanospora, Syspastospora and the species previously placed in
Persiciospora and Sphaerodes has already been demonstrated by numerous authors (Doguet
1955, Calviello 1973, Jordan and Barnett 1978, Harveson and Kimbrough 2000, 2001),
and this ability has been exploited in the biocontrol of phytopathogenic fungi (Vujanovic
and Goh 2009, Goh and Vujanovic 2010, Kim and Vujanovic 2016, 2017).
The genus Sphaeronaemella, which is characterized by pale and translucent
ascomata, was thought to be related to Melanospora (Cannon and Hawksworth
1982). However, we do not agree with this relationship because it differs from the
Melanosporales in the production of hyaline ascospores, as opposed to the pigmented
ones in the members of that order. By contrast, our results correlate with those of other
authors that demonstrated a closer phylogenetic relationship of this genus with the
members of the order Microascales (Spatafora and Blackwell 1994b, Hausner and Reid
2004). In fact, our SSU tree seems to indicate that Sphaeronaemella could represent
a new family of the Microascales; however, further studies including more taxa and
additional genes are needed to more accurately confirm its taxonomic status.
The placement of our isolate of Persiciospora japonicum in the Microthecium clade
once more demonstrated that the ornamentation of the ascospores, which is pitted
in Persiciospora spp., is of poor taxonomic value, and consequently all the species of
Persiciospora should be transferred to Microthecium. As it was above mentioned, the
species of this latter genus show a typical cellular ascomatal neck which is also present
in Persiciospora and constitutes a common feature in both genera. Surprisingly, in some
previous phylogenetic studies, the species of Persiciospora were placed in the Hypocreales,
closely related to Nectria (Zhang and Blackwell 2002, Maharachchikumbura et al.
2015, Schultes et al. 2017). However, this could be probably explained by a possible
contamination of the cultures of Persiciospora spp. with an hypocrealean host (Fan
et al. 2012). The same situation may have occurred with the cultures of Scopinella
and Syspastospora, which led to a possible erroneous classification of both taxa in the
Melanospora (Sordariomycetes, Ascomycota) and its relatives
117
Hypocreales (Zhang and Blackwell 2002, Chaudhary et al. 2006, Fan et al. 2012,
Maharachchikumbura et al. 2015, Schultes et al. 2017).
Pteridiosperma ciliatum, a member of the Melanosporales with ascospores
ornamented with longitudinal wing-like ridges that anastomose each other to form
a well defined reticulum (a distinctive feature of Pteridiosperma), was also found
in the Microthecium clade, proving once again that the ascospore ornamentation is
not phylogenetically informative. Consequently, we have synonymyzed the genus
Pteridiosperma with Microthecium since Pteridiosperma spp. show non-ostiolate
ascomata, or if ostiolate, they show a short neck composed of angular cells, which are
typical morphological characteristics of Microthecium.
Another genus that our results demonstrated should be synonymized and included in
Microthecium is Sphaerodes because its type species, S. episphaerium, shows morphological
features (non-ostiolate ascomata) that fit with the current circumscription of that
emended genus. Most of the species of Sphaerodes, with the exception of S. ellipsospora
and S. singaporensis, which are now located in the new genus Dactylidispora, and S.
mycoparasitica, which is now placed in Melanospora, are also transferred to Microthecium
since these produce non-ostiolate or ostiolate ascomata without a neck, or less frequently
with a short neck composed of angular cells similar to the ascomatal ones. Another
relevant feature of the genus Microthecium is the production of bulbils. These propagules
are typical of Papulaspora, an anamorphic genus that encompasses more than 40 species.
Although it was initially accepted as a genus without a sexual morph (Hotson 1912),
its link with species of Melanospora and Chaetomium has been reported (Roll-Hansen
1948, Zhang et al. 2004). In our phylogenetic study Papulaspora sepedonioides, the type
species of the genus, was nested in the Microthecium clade, and therefore transferred
to this genus. The relationship of this species with the Melanosporales had already
previously been demonstrated by Davey et al. (2008) and Li et al. (2016). However, it
has been demonstrated that Papulaspora is a polyphyletic genus, and other species of the
genus have been reported as belonging to the classes Leotiomycetes and Sordariomycetes
(Ascomycota). Therefore, the other species of Papulaspora not linked to the species of
Microthecium should be transferred to other taxonomic groups. The relationship of
some species of Papulaspora with the Melanosporales is also suggested by the production
of similar phialidic synanamorphs (Van Beyma 1931, Hotson 1942).
The most recent new combination performed in Sphaerodes, S. inferior, was done
to accommodate S. retispora var. inferior since it was not clustering with S. retispora var.
retispora (Schultes et al. 2017). However, we suspected that the sequences of S. retispora var.
retispora deposited in GenBank were contaminated with the hypocrealean host. In order
to corroborate it, we studied such sequenced strain demonstrating that it was effectively
contaminated. Therefore, S. inferior is here considered a synonym of Mi. retisporum since
the morphological differences are insufficient to recognize this variety as a different species.
There are important morphological differences among the strains of Microthecium
that suggest the presence of several additional cryptic species in the genus; however,
our phylogenetic study, in spite of having used five loci, was not able to resolve the
boundaries among them.
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Acknowledgments
This work was supported by the Spanish “Ministerio de Economía y Competitividad”,
grant CGL2017-88094-P. The new genus Echinusitheca was recovered from samples
collected during fieldwork supported by a National Science Foundation grant (DEB0515558) to ANM.
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