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ORIGINAL ARTICLE
Phylogenetic placement of Leptosphaeria polylepidis, a pathogen
of Andean endemic Polylepis tarapacana, and its newly discovered
mycoparasite Sajamaea mycophila gen. et sp. nov.
Marcin Piątek
1
&Pamela Rodriguez-Flakus
2
&Alejandra Domic
3,4
&Arely N. Palabral-Aguilera
5
&
M. Isabel Gómez
6
&Adam Flakus
7
Received: 12 April 2019 /Revised: 15 October 2019 /Accepted: 21 October 2019
Abstract
Polylepis tarapacana forms one of the highest-altitude woodlands worldwide. Its populations are experiencing a decline due to
unsustainable land-use practices, climate change, and fungal infection. In Sajama National Park in Bolivia, Polylepis tarapacana
is affected by a disease caused by the pleosporalean fungus Leptosphaeria polylepidis, recently described in 2005. In this study,
the integrative morphological and molecular analyses using sequences from multiple DNA loci showed that it belongs to the
genus Paraleptosphaeria (Leptosphaeriaceae, Pleosporales). Accordingly, the appropriate new combination, Paraleptosphaeria
polylepidis, is made. Pseudothecia of Pa. polylepidis were found to be overgrown by enigmatic conidiomata that were not
reported in the original description of this fungus. Morphological and molecular analyses using sequences from two DNA loci
revealed that they belong to an undescribed genus and species in the family Dictyosporiaceae (Pleosporales). The new generic
and specific names, Sajamaea and S. mycophila, are introduced for this unusual fungus.
Keywords Andes .Mycoparasite .New genus .New species .Pleosporales .Plant pathogen .Polylepis .South America
Introduction
Along the Andes, dispersed woodlands of Polylepis
(Rosaceae) constitute a common component of the treeline.
Polylepis woodlands are characterized by being dominated
either mostly or exclusively by representatives of this genus
and by being found in areas of difficult access, including
rocky outcrops and mountain slopes (Fjeldså and Kessler
1996; Kessler 2006). These woodlands are important habitats
for plant and animal species, including several endangered
habitat-specialist bird species (Fjeldså and Kessler 1996)and
constitute important sources of firewood, fodder, and medici-
nal plants for local indigenous communities (Fjeldså and
Kessler 1996;Domicetal.2014; Hurtado et al. 2018).
Polylepis woodlands are experiencing a rapid decline due to
unsuitable land use practices, habitat destruction, and ongoing
climate change (Navarro et al. 2005).
Polylepis tarapacana is a species that includes small trees
and shrubs distributed along the semiarid Andean highlands
from Peru to Argentina and Chile (Kessler 1995). The species
Section Editor: Hans-Josef Schroers
*Marcin Piątek
m.piatek@botany.pl
*Pamela Rodriguez-Flakus
p.rodriguez@botany.pl
1
Department of Mycology, W. Szafer Institute of Botany, Polish
Academy of Sciences, Lubicz 46, 31-512 Kraków, Poland
2
Department of Vascular Plants, W. Szafer Institute of Botany, Polish
Academy of Sciences, Lubicz 46, 31-512 Kraków, Poland
3
Department of Anthropology, The Pennsylvania State University,
University Park, PA 16802, USA
4
Department of Geosciences, The Pennsylvania State University,
University Park, PA 16802, USA
5
Instituto de Ecología, Herbario Nacional de Bolivia, Calle 27 Cota
Cota,LaPaz,Bolivia
6
Colección Boliviana de Fauna, Museo Nacional de Historia Natural,
La Paz, Bolivia
7
Department of Lichenology, W. Szafer Institute of Botany, Polish
Academy of Sciences, Lubicz 46, 31-512 Kraków, Poland
Mycological Progress (2020) 19:1–14
https://doi.org/10.1007/s11557-019-01535-w
#The Author(s) 2019
possess morphological (Simpson 1979; Kessler 1995) and
physiological adaptations (Rada et al. 2001; Azócar et al.
2007; González et al. 2007) to tolerate frost and desiccation
due to extreme environmental conditions, including high solar
irradiance, night frosts, and high diurnal temperature varia-
tion. In Nevado Sajama in Bolivia, P. tarapacana forms one
of the world’s highest-altitude woodlands, reaching elevations
up to 5200 m a.s.l. (Jordan 1980). The species is currently
categorized as “Vul ner able”in Bolivia due to firewood extrac-
tion, habitat loss, and climate change (MMAyA 2012;
Cuyckens et al. 2016). A potentially pathogenic fungus,
Leptosphaeria polylepidis, constitutes an emerging threat to
the permanence of remaining P. tarapacana woodlands, as its
infection has been attributed to the mortality of several indi-
viduals in the Sajama National Park (Coca-Morante 2012).
The fungus, originally described from two collections made
in 2002 in the Sajama National Park, forms stromatic black
knots on the branches of infected individuals of P. tarapacana
(Macía et al. 2005). Recent evaluations have shown that the
infection is widespread in protected areas particularly of the
southeast and northeast slopes of Nevado Sajama. However,
actions to control the spread of the infection have not been
implemented due to a lack of knowledge regarding the life-
cycle and pathogenicity of L. polylepidis.
The genus Leptosphaeria, with the type species
Leptosphaeria doliolum, resides in the Leptosphaeriaceae
within Pleosporales (Zhang et al. 2012; de Gruyter et al.
2013;Ariyawansaetal.2015). It contains hundreds of de-
scribed species that in recent molecular analyses were proven
to belong to different genera, including Brunneosphaerella,
Heterospora,Neoleptosphaeria,Paraleptosphaeria,
Plenodomus,Pseudoleptosphaeria,orSubplenodomus
(Crous et al. 2009; de Gruyter et al. 2013;Ariyawansaetal.
2015). The generic placement of the majority of
Leptosphaeria species is, however, still uncertain, and their
rearrangements into the correct genus need molecular
phylogenetic analyses, ideally using freshly collected
materials. When describing Leptosphaeria polylepidis,
Macía et al. (2005) provided a nuc rDNA ITS1-5.8S-ITS2
(ITS) sequence from the holotype material. Comparing this
sequence with sequences available at that time in GenBank,
they found that L. polylepidis was most similar to
Leptosphaeria dryadis, a parasite of Dryas octopetala in arctic
and alpine regions of the Northern Hemisphere.
Leptosphaeria dryadis is currently placed in the genus
Paraleptosphaeria (de Gruyter et al. 2013). However, it is
not clear if this is also the correct genus for L. polylepidis
because the ITS sequence generated by Macía et al. (2005)
was not included in any of the subsequent studies dealing with
Leptosphaeria-like species; many new sequences of
Leptosphaeria-like species were generated in recent years that
could disprove close relationships between L. polylepidis and
Paraleptosphaeria dryadis, and sequences from more than
one DNA locus can better resolve generic boundaries within
Leptosphaeriaceae (Ariyawansa et al. 2015).
Therefore, it is the aim of this study to resolve the phyloge-
netic and generic placement of Leptosphaeria polylepidis by
employing integrative morphological and molecular analyses
using sequences from multiple DNA loci. During the analyses
of freshly collected material, we unexpectedly found that
pseudothecia of L. polylepidis were overgrown by conidiomata
that were not reported in the original description of the fungus.
It was not clear if these conidiomata represented an asexual
state of L. polylepidis or, alternatively, belonged to a
mycoparasitic fungus. Therefore, the second aim of this study
was to answer this question by applying morphological and
molecular analyses using nuc rDNA ITS and LSU sequences.
Materials and methods
Specimen sampling and documentation
This study is based on newly collected material of
Leptosphaeria polylepidis. Four specimens were collected in
the Sajama National Park in Bolivia, which is the type locality
and the only known place of occurrence of L. polylepidis.One
of them was partly covered with conidiomata. This specimen
was used for detailed morphological/anatomical and molecu-
lar analyses. The voucher material is preserved at the Herbario
Nacional de Bolivia (LPB) and at the fungal herbarium of the
W. Szafer Institute of Botany, Polish Academy of Sciences,
Kraków (KRAM F). Attempts to obtain cultures failed.
Morphological analyses
The morphology was examined using standard stereo and
light microscopy (Nikon SMZ 800, Nikon Eclipse 80i DIC).
Thin sections of ascomata and conidiomata were made man-
ually using a razor blade or with the aid of a freezing micro-
tome Thermo Scientific Microm HM430 equipped with BFS-
MP freezing stage and BFS-3MP controller. The sectioned
material was examined in distilled water, 10% solution of
potassium hydroxide (KOH), or lactophenol cotton blue
(LPCB). The amyloidity of fungal structures was tested using
Lugol’s solution (IKI), or a combination of first KOH and then
IKI (KOH/IKI). All measurements were made in distilled
water.
DNA isolation, PCR, and sequencing
Genomic DNA was extracted in three separate extraction
rounds, namely one extraction from 10 pseudothecial hymenia
of Leptosphaeria polylepidis, one extraction from conidial
mass obtained from five conidiomata, and one extraction from
15 entire conidiomata of unknown fungus, using the
2 Mycol Progress (2020) 19:1–14
DNeasy™Plant Mini Kit or QIAamp DNA Investigator Kit
(Qiagen, Germany), following the manufacturer’sinstruc-
tions. For Leptosphaeria polylepidis, a total of four genetic
markers were amplified, namely nuc rDNA 18S (SSU), nuc
rDNA ITS1-5.8S-ITS2 (ITS), nuc rDNA 28S (LSU), and a
fragment of the translation elongation factor 1-alpha gene
(TEF1). For unknown conidiomatal fungus, two genetic
markers were amplified, namely ITS and LSU. The following
primer pairs were used for amplification: NS1-NS24 (White
et al. 1990; Gargas and Taylor 1992), ITS1F-ITS4 (White
et al. 1990; Gardes and Bruns 1993), LROR-LR7 (Vilgalys
and Hester 1990; Rehner and Samuels 1994), and EF1983F-
EF12218R (Rehner and Buckley 2005) for SSU, ITS, LSU,
and TEF1, respectively. Polymerase chain reactions (PCRs)
were performed as follows: for Leptosphaeria polylepidis,
2μl of DNA extract was used for SSU, ITS, and LSU and
5μlforTEF1, while for unknown conidiomatal fungus, 3 μl
of DNA extract was used for ITS and LSU in a total volume of
25 μl PCR reactions; the rest of components were added ac-
cording to Flakus et al. (2019). Cycling conditions were per-
formed as reported by Rodriguez-Flakus and Printzen (2014)
for the nuclear markers and Rehner and Buckley (2005)for
TEF1 with modification of the initial denaturalization to 4 min
and final extension to 5 min (M. Mardones, pers. comm.).
PCR products were visualized on agarose gels and later puri-
fied using ExoSap. The PCR amplicons were sequenced by
Macrogen Europe B.V. (Amsterdam, the Netherlands).
Phylogenetic analyses
Newly generated sequences were trimmed, assembled, and
edited using Geneious Pro, version 8.0.5 (Biomatters Inc.).
BLAST searches in GenBank (Altschul et al. 1997) were per-
formed to find sequences of most closely related species, re-
vealing affinities of sequences from Leptosphaeria polylepidis
to sequences of the Leptosphaeriaceae and sequences obtained
from unknown conidiomatal fungus to sequences of the
Dictyosporiaceae. Accordingly, two separate datasets were as-
sembled to resolve the phylogenetic placement of each fungus.
To resolve the phylogenetic position of Leptosphaeria
polylepidis, a four-gene dataset (SSU+ITS+LSU+TEF1)was
assembled that included sequences generated in this study and
sequences from 45 specimens of closely related members of
the Leptosphaeriaceae and a member of the Cucurbitariaceae
used as an outgroup (selected from Zhang et al. 2012 and
Tibpromma et al. 2017;seeTable1). Each single gene dataset
was aligned separately using the MAFFT algorithm (Katoh
et al. 2005) as implemented on the GUIDANCE web-server
(Penn et al. 2010); to remove poorly or ambiguously aligned
uncertain columns, a default cut-off score of 0.93 in all single
gene alignments were chosen. The single-gene datasets were
concatenated to a final alignment using Geneious Pro and
consisted of 1023 bp (SSU), 452 bp (ITS), 1326 bp (LSU),
To resolve phylogenetic position of the conidiomatal fungus,
a two-gene dataset (ITS+LSU) was assembled, which
contained sequences generated in this study and sequences
from 28 related members of the Dictyosporiaceae and two se-
quences of Periconia igniaria used as an outgroup (selected
from Iturrieta-González et al. 2018 and Yang et al. 2018;see
Tab le 2). Phylogenetic analyses were performed as described
above and were based on 820 bp (LSU) and 412 bp (ITS)
alignments. A single partition for LSU (GTR+I+G) and three
partitions for ITS (GTR+G for ITS1, SYM+I+G for 5.8S, and
JC+I+G for ITS2) were selected based on PartitionFinder 2
(Lanfear et al. 2016). The ML and BI analyses were performed
with similar parameters as described above, but 20 million gen-
erations in four independent parallel runs were sampled and six
gradually heated chains used. Relationships receiving bootstrap
support (ML-BP) above 70% and 0.95 as Bayesian posterior
probability (PP) were considered strongly supported. The
resulting trees from both phylogenetic analyses were built in
Figtree v.1.4.2 (http//tree.bio.ed.ac.uk/software/figtree).
Results
Phylogenetic analyses
The phylogenetic analyses of the four-locus (SSU+ITS+LSU+
TEF1) dataset comprising representatives of the
Leptosphaeriaceae (and a member of the Cucurbitariaceae used
as an outgroup) resulted in overall similar tree topologies
Mycol Progress (2020) 19:1–14 3
and 915 bp (TEF1). Subsequently, the analyses were per-
formed in the CIPRES Scientific gateway portal (:dito_
existshttp://www.phylo.org/portal2) (Miller et al. 2010). The
optimal partitioning scheme and substitution models for each
single-gene alignment were inferred by PartitionFinder 2
(Lanfear et al. 2016). A single partition for SSU and LSU;
three partitions for ITS1, 5.8S, and ITS2 regions; and three
for each of the codon positions of TEF1 were found.
Substitution models selected under the greedy search algo-
rithm and Akaike information criterion (Lanfear et al. 2016)
were TVM+G+I for SSU, ITS2, and LSU; JC+I for 5.8S; and
GTR+G for ITS1 and used as priors for Bayesian phylogenet-
ic inference (BI) analyses. Maximum likelihood (ML) and
bootstrap support (BS) analyses were performed on each locus
separately and concatenated data set by using RAxML-HPC2
on XSEDE with a rapid BS algorithm (Stamatakis 2014),
1000 replicates, and GTRGAMMA substitution model.
Bayesian inference (BI) analyses were carried out using
Markov Chain Monte Carlo as implemented in MrBayes on
XSEDE (Huelsenbeck and Ronquist 2001; Ronquist and
Huelsenbeck 2003) and were based on 10 million generations,
sampling every 1000th tree, two independent parallel runs of
four incrementally heated chains (0.15) and discarding the
first 0.5 of the sampled trees.
Table 1 Species used for inferring the phylogeny of the Leptosphaeriaceae (and a member of the Cucurbitariaceae used as an outgroup) with details of
their host, country of origin, strain or voucher information, GenBank accession numbers (ITS, LSU, SSU, TEF1)andreferences
Species Host Country Strain/
voucher
GenBank acc. no. References
ITS LSU SSU/TEF1
Alternariaster
bidentis
Bidens
sulphurea
Brazil CBS
134021
KC609333 KC609341 –/–Alves et al. 2013
Alternariaster
bidentis
Bidens
sulphurea
Brazil CBS
134185
KC609334 KC609342 –/–Alves et al. 2013
Alternariaster
centaureae--
diffusae
Centaurea
diffusa
Russia MFLUCC
14-0992
KT454723 KT454715 KT454730/–Ariyawansa et al. 2015
Alternariaster
centaureae--
diffusae
Centaurea
diffusa
Russia MFLUC
15-0009
KT454724 KT454716 KT454731/–Ariyawansa et al. 2015
Alternariaster
helianthi
Helianthus
annuus
Hungary CBS
199.86
KC609336 KC609343 –/–Alves et al. 2013
Alternariaster
helianthi
Helianthus
annuus
–CBS
327.69
KC609335 KC584369 KC584627/–Alves et al. 2013
Cucurbitaria
berberidis
Berberis
julianae
Netherlands CBS
394.84
–JX681088 GQ387544/–Ve r k l e y e t a l . 2014 (ITS); de Gruyter
et al. 2010 (LSU)
Heterospora
chenopodii
Chenopodium
album
Netherlands CBS
115. 96
JF740227 EU754188 EU754089/–de Gruyter et al. 2013 (ITS); de
Gruyter et al. 2009 (LSU, SSU)
Heterospora
chenopodii
Chenopodium
album
Netherlands CBS
448.68
FJ427023 EU754187 EU754088/–Aveskamp et al. 2009 (ITS); de
Gruyter et al. 2009 (LSU, SSU)
Heterospora
dimorphospora
Chenopodium
quinoa
Peru CBS
165.78
–JF740281 JF740098/–de Gruyter et al. 2013
Heterospora
dimorphospora
Chenopodium
quinoa
Peru CBS
345.78
–GU238069 GU238213/–Aveskamp et al. 2010
Leptosphaeria
doliolum
dead stem United
Kingdo-
m
MFLUC
15-1875
KT454727 KT454719 KT454734/–Ariyawansa et al. 2015
Leptosphaeria
doliolum
Phlox
paniculata
Netherlands CBS
155.94
JF740207 JF740282 –/–de Gruyter et al. 2013
Leptosphaeria
doliolum
Urtica dioica Netherlands CBS
505.75
JF740205 GQ387576 GQ387515/GU349069 de Gruyter et al. 2013 (ITS); de
Gruyter et al. 2010 (LSU, SSU);
Schoch et al. 2009 (TEF1)
Leptosphaeria
doliolum
Rudbeckia Netherlands CBS
541.66
JF740206 JF740284 –/–de Gruyter et al. 2013
Leptosphaeria
errabunda
Solidago Netherlands CBS
617.75
JF740216 JF740289 –/–de Gruyter et al. 2013
Leptosphaeria
errabunda
Delphinium Netherlands CBS
125978
JF740217 JF740290 –/–de Gruyter et al. 2013
Leptosphaeria
pedicularis
Pedicularis Switzerland CBS
390.80
JF740224 JF740294 –/–de Gruyter et al. 2013
Leptosphaeria
pedicularis
Gentiana
punctata
Switzerland CBS
126582
JF740223 JF740293 –/–de Gruyter et al. 2013
Leptosphaeria
sclerotioides
Medicago
sativa
Canada CBS
144.84
JF740192 JF740269 –/–de Gruyter et al. 2013
Leptosphaeria
sclerotioides
Medicago
sativa
Canada CBS
148.84
JF740193 JF740270 –/–de Gruyter et al. 2013
Leptosphaeria
slovacica
Ballota nigra Netherlands CBS
389.80
JF740247 JF740315 JF740101/–de Gruyter et al. 2013
Leptosphaeria
slovacica
Ballota nigra Netherlands CBS
125975
JF740248 JF740316 –/–de Gruyter et al. 2013
Leptosphaeria
sydowii
Senecio
jacobaea
United
Kingdo-
m
CBS
385.80
JF740244 JF740313 –/JF740139 de Gruyter et al. 2013
Leptosphaeria
sydowii
Senecio
jacobaea
United
Kingdo-
m
CBS
125976
JF740245 JF740314 –/–de Gruyter et al. 2013
Paraleptosphaeria
dryadis
Dryas
octopetala
Switzerland CBS
643.86
JF740213 GU301828 KC584632/GU349009 de Gruyter et al. 2013 (ITS); Schoch
et al. 2009 (LSU, TEF1);
Woudenberg et al. 2013 (SSU)
Paraleptosphaeria
dryadis
Dryas
octopetala
Switzerland CBS
743.86
AF439461 ––/–Camara et al. 2002
4 Mycol Progress (2020) 19:1–14
obtained from BI and ML analyses. The Heterospora clade
nested together with Subplenodomus in the ML analysis but
was sister to the remaining genera of the Leptosphaeriaceae in
BI analysis. In general, high bootstrap support values above
70% (ML–BP) and posterior probabilities above 0.95 (PP) were
retrieved and supported monophyletic lineages representing
distinct genera as in previous studies (de Gruyter et al. 2013;
Ariyawansa et al. 2015; Tibpromma et al. 2017). The Bayesian
tree is displayed in Fig. 1. The newly generated Leptosphaeria
polylepidis sequences clustered with ex-holotype sequence of
L. polylepidis (ML–BP = 100%, PP = 1) in their well-supported
Paraleptosphaeria lineage (ML–BP = 78%, PP = 1);
Tabl e 1 (continued)
Species Host Country Strain/
voucher
GenBank acc. no. References
ITS LSU SSU/TEF1
Paraleptosphaeria
macrospora
Rumex
longifolius
Norway CBS
114198
JF740238 JF740305 –/–de Gruyter et al. 2013
Paraleptosphaeria
nitschkei
Cirsium
spinosissim-
um
Switzerland CBS
306.51
NR_111 621
(=J-
F740239)
JF740308 –/–de Gruyter et al. 2013
Paraleptosphaeria
nitschkei
Petasites Italy MFLU
13-0644
KP729446 KP729447 –/–Liu et al. 2015
Paraleptosphaeria
polylepidis
Polylepis
tarapacana
Bolivia MA-Fungi
57843 –
holotype
AJ786644 ––/–Macia et al. 2005
Paraleptosphaeria
polylepidis
Polylepis
tarapacana
Bolivia APA-2999 MK795714 MK795717 MK795720/MK831009 This study
Paraleptosphaeria
praetermissa
Rubus idaeus Sweden CBS
114591
JF740241 JF740310 –/–de Gruyter et al. 2013
Paraleptosphaeria
rubi
Rubus Italy MFLUCC
14-0211
KT454726 KT454718 KT454733/–Ariyawansa et al. 2015
Paraleptosphaeria
sp. (as
Leptosphaeria
sp.)
Phlomis
younghusba-
ndii
China PHY-06 JX401979 JX401985 –/–Zhang D.W. and Guo S.X. unpubl.
Paraleptosphaeria
sp. (as
Leptosphaeria
sp.)
Phlomis
younghusba-
ndii
China PHY-54 JX401931 JX401999 –/–Zhang D.W. and Guo S.X. unpubl.
Plenodomus
biglobosus
Brassica juncea France CBS
127249
JF740199 JF740275 –/–de Gruyter et al. 2013
Plenodomus
biglobosus
Brassica napus
var.
napobrassic-
a
–CBS
298.36
AJ550862 GU237980 GU238207/–Mendes-Pereira et al. 2003 (ITS);
Aveskamp et al. 2010 (LSU, SSU)
Plenodomus lupini Lupinus
mutabilis
Peru CBS
248.92
JF740236 JF740303 –/–de Gruyter et al. 2013
Plenodomus
pimpinellae
Pimpinella
anisum
Israel CBS
101637
JF740240 JF740309 –/–de Gruyter et al. 2013
Plenodomus
salviae
Salvia glutinosa Italy MFLUCC
13-0219
KT454725 KT454717 KT454732/–Ariyawansa et al. 2015
Plenodomus sp. (as
Leptosphaeria
sp.)
Phlomis
younghusba-
ndii
China PHY-30 JX401955 JX401989 –/–Zhang D.W. and Guo S.X. unpubl.
Plenodomus visci Viscum album France CBS
122783
–EU754195 EU754096/–de Gruyter et al. 2009
Plenodomus
wasabiae
Eutrema
japonicum
Taiwan CBS
120119
JF740257 JF740323 –/–de Gruyter et al. 2013
Plenodomus
wasabiae
Eutrema
japonicum
Taiwan CBS
120120
JF740258 JF740324 –/–de Gruyter et al. 2013
Subplenodomus
violicola
Viola tricolor Netherlands CBS
306.68
FJ427083 GU238156 –/–Aveskamp et al. 2009 (ITS);
Aveskamp et al. 2010 (LSU)
Subplenodomus
violicola
Viola tricolor New
Zealand
CBS
100272
FJ427082 JF740322 –/–Aveskamp et al. 2009 (ITS); de
Gruyter et al. 2013 (LSU)
APA A. N. Palabral-Aguilera; CBS CBS-KNAW Collections, Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; MA Real Jardín
Botánico herbarium, Madrid, Spain; MFLU Mae Fah Luang University herbarium; MFLUCC Mae Fah Luang University Culture Collection, Chiang
Rai, Thailand
Mycol Progress (2020) 19:1–14 5
relatedness of L. polylepidis and Paraleptosphaeria dryadis
was weakly supported (ML–BP = 60%). The ITS sequence
from the newly generated material and holotype of
L. polylepidis differed in 6 bp. The SSU, LSU, and TEF1 se-
quences were not available from holotype material.
The phylogenetic analyses of the two-locus (ITS+LSU)
dataset, including representatives of the Dictyosporiaceae and
two sequences of Periconia igniaria used as an outgroup, re-
sulted in similar tree topologies. Dictyosporium grouped in a
single clade together with Aquadictyospora in ML analysis but
was sister to the Pseudodictyosporium lineage in the BI analy-
sis. The Bayesian consensus tree is displayed in Fig. 2.The
phylogenetic tree shows 11 monophyletic lineages of the
Dictyosporiaceae, corresponding to 10 known genera and one
unknown lineage, the latter of which consisted of the
conidiomatal fungus from Leptosphaeria polylepidis.Results
were roughly consistent with previous phylogenetic results
(e.g., Iturrieta-González et al. 2018;Yangetal.2018), except
for some differences in branches with low statistical support
values mainly at the backbone of the phylogenetic tree. The
sequences of the conidiomatal fungus from two independent
extraction rounds were identical and of high quality, excluding
the possibility of DNA isolation from contaminating fungi, and
were nested within the currently circumscribed family
Dictyosporiaceae (ML–BP=100%,PP=1).Theyformeda
sister group to the members of the genus Pseudocoleophoma
(ML–BP = 89%, PP = 1). The phylogenetic relationships of the
conidiomatal fungus and the Pseudocoleophoma lineage with
their relatives in the Dictyosporiaceae remain unresolved, due
the lack of support in our molecular analyses.
Taxonomy
Paraleptosphaeria polylepidis (M.J. Macía, M.E. Palm &
M.P. Martín) Piątek, Flakus & Rodr. Flakus, comb. nov.
(Fig. 3)
MycoBank no. MB832195
Basionym:Leptosphaeria polylepidis M.J. Macía, M.E.
Palm & M.P. Martín, Mycotaxon 93: 402 (2005)
Ascomata pseudothecial, developedonstroma.Stroma 1–3
mm high, 2–7 mm wide, black, surface with visible individual
pseudothecia, usually with base growing inward and intermixed
with host tissue. Pseudothecia dark brown to black, matt, rough,
300–700 μmhigh,300–600 μm wide, globose to subglobose
with base usually protruding into ca. 200–1000 μmhighstipes
that fuse below into stroma; papillae short or lacking. Peridium
60–150 μm wide, pale to dark brown (KOH+ greenish grey),
consisting of a single stratum, paraplectenchymatous, textura
angularis, composed of several layers of isodiametric to slightly
elongate thin-walled cells, 15–25 × 9–15 μm. Hamathecium
persistent, composed of relatively thin, 1.5–4μm wide, septate,
branched and strongly anastomosed pseudoparaphyses. Asci
120–250 × 20–45 μm, 5–8-spored, bitunicate, cylindrical-
clavate, slightly curved, with a short stipe, apically widened,
with an ocular chamber, IKI–, KOH/IKI–; endosacus KOH/
IKI+ orange. Ascospores 45–60 × (14–)20–23 μm, pale brown,
broadly ellipsoidal, narrower at the top, constricted at septa,
uniformly thin-walled, without perispore or gelatinous coat,
(2–)3(–4)-septate. Asexual state unknown.
Specimen examined (used for morphology and molecular
analyses): See host fungus in the type of Sajamaea mycophila.
Additional specimens examined: Bolivia, Oruro, Sajama
National Park (=Parque Nacional Sajama), W slope of
Sajama’s volcano, Quebrada Huaytana, 18°11′S, 68°51′W,
elev. ca. 4000–4200 m, on Polylepis tarapacana, 1 Feb. 2016,
A. Domic (three collections: AD1, AD2, AD5; all in LPB).
Sajamaea Flakus, Piątek & Rodr. Flakus, gen. nov.
MycoBank no. MB832196
Etymology: Referring to the place of occurrence of the new
genus –slopes of Nevado Sajama, the highest mountain in
Bolivia.
Description:Mycoparasitic.Conidiomata pycnidial,
uniloculate to multi-loculate, subglobose to irregular in shape,
pale brown. Peridium composed of several layers of isodia-
metric to slightly elongate cells in the form of textura
angularis. Conidiophores hyaline, reduced to phialidic
conidiogenous cells. Conidia pigmented (pale brown), broad-
ly ellipsoidal, aseptate, smooth, thin-walled, guttulate.
Generic type:Sajamaea mycophila Flakus,M.Piątek &
Rodr. Flakus.
Sajamaea mycophila Flakus, Piątek & Rodr. Flakus, sp.
nov. (Fig. 4)
MycoBank no. MB832197
Etymology: The epithet name mycophila refers to the oc-
currence of this fungus on other fungus.
Description:Mycoparasitic on Paraleptosphaeria
polylepidis, causing moderate damages of host ascomata.
Conidiomata pycnidial, as pale brown galls on host surface,
150–300 μm wide, 120–300 μm high, first erumpent through
the outermost layer of host peridium, later almost sessile on
pseudothecial clusters of host, uniloculate to multi-loculate,
solitary to aggregated, subglobose to irregular in shape, pale
brown; better seen when wet. Peridium 20–30 μm wide, pale
to dark brown, composed of about 3–8 layers of cells with
walls up to 2 μm wide, cells isodiametric to slightly elongate,
textura angularis. Conidiophores reduced to conidiogenous
cells. Conidiogenous cells 5–11 μmhigh,4–8μmwide,
enteroblastic, phialidic, smooth-walled, hyaline, with a small
collar, densely outlining inner surfaces of pycnidia. Conidia
9–13 × 5.5–7.5 μm, pale brown, broadly ellipsoidal, sometimes
slightly narrower at one point, with rounded ends, aseptate,
smooth, thin-walled, guttulate. Sexual state unknown.
Specimen examined: Bolivia, Oruro, Sajama National Park
(=Parque Nacional Sajama), W slope of Nevado Sajama,
6 Mycol Progress (2020) 19:1–14
Table 2 Species used for inferring phylogeny of the Dictyosporiaceae (and Periconia igniaria used as an outgroup) with details of their substrate or
host, country of origin, strain or voucher information, GenBank accession numbers (ITS, LSU), and references
Species Substrate/host Country Strain/voucher GenBank acc. no. References
ITS LSU
Aquadictyospora
lignicola
Submerged wood China MFLUCC 17-1318 MF948621 MF948629 Li et al. 2017
Cheirosporium
triseriale
Submerged wood China HMAS 180703 EU413953 EU413954 Cai et al. 2008
Dendryphiella
paravinosa
Leaves of Citrus
sinensis
Italy CBS 14128 KX228257 KX228309 Crous et al. 2016
Dendryphiella
variabilis
Dead leaf of a
lauraceous tree
Cuba CBS 584.96 LT963453 LT963454 Iturrieta-González et al.
2018
Dendryphiella vinosa –Japan (for ITS,
unavailable
for LSU)
NBRC 32669 (for ITS,
strain info unavailable
for LSU)
DQ307316 EU848590 Dela Cruz T.E. et al. unpubl.
(ITS); Jones et al. 2008
(LSU)
Dictyocheirospora
aquatica
Submerged wood China KUMCC 15-0305 KY320508 KY320513 Wang et al. 2016
Dictyocheirospora
bannica
Dead wood Japan KH 332 LC014543 AB807513 Tanaka et al. 2015
Dictyocheirospora
rotunda
Submerged dead
decaying wood
Thailand MFLUCC 14-0293 KU179099 KU179100 Boonmee et al. 2016
Dictyocheirospora
rotunda
Submerged wood China MFLUCC 17-1313 (in
GenBank) or
MFLUCC 17–1687 (in
publication)
MF948625 MF948633 Li et al. 2017
Dictyosporium
bulbosum
Dead wood Japan yone 221 LC014544 AB807511 Tanaka et al. 2015
Dictyosporium
elegans
––NBRC 32502 DQ018087 DQ018100 Tsui et al. 2006
Dictyosporium
thailandicum
Submerged wood Thailand MFLUCC 13-0773 KP716706 KP716707 Liu et al. 2015
Dictyosporium
toruloides
––CBS 209.65 DQ018093 DQ018104 Tsui et al. 2006
Digitodesmium
bambusicola
––CBS 110279 DQ018091 DQ018103 Tsui et al. 2006
Gregarithecium
curvisporum
Dead culms of Sasa
sp.
Japan KT 922 AB809644 AB807547 Tanaka et al. 2015
Jalapriya inflata Piece of driftwood United Kindom NTOU 3855 JQ267362 JQ267363 Kirschner et al. 2013
Jalapriya pulchra Submerged decaying
wood
China MFLUCC 15-0348 KU179108 KU179109 Boonmee et al. 2016
Jalapriya pulchra Submerged wood China MFLUCC 17-1683 MF948628 MF948636 Li et al. 2017
Neodendryphiella
mali
Leaf of Malus
domestica
Italy CBS 139.95 LT906655 LT906657 Iturrieta-González et al.
2018
Neodendryphiella
mali
Herbivore dung Spain FMR 17003 LT993734 LT993735 Iturrieta-González et al.
2018
Neodendryphiella
michoacanensis
Soil Mexico FMR 16098 LT906660 LT906658 Iturrieta-González et al.
2018
Neodendryphiella
tarraconensis
Soil Spain FMR 16234 LT906659 LT906656 Iturrieta-González et al.
2018
Periconia igniaria Triticum aestivum Switzerland CBS 379.86 LC014585 AB807566 Tanaka et al. 2015
Periconia igniaria Bamboo Papua New
Guinea
CBS 845.96 LC014586 AB807567 Tanaka et al. 2015
Pseudocoleophoma
calamagrostidis
Dead leaves of
Calamagrostis
matsumurae
Japan KT 3284 LC014592 LC014609 Tanaka et al. 2015
Pseudocoleophoma
polygonicola
Dead stems of
polygonaceous
plant
Japan KT 731 AB809634 AB807546 Tanaka et al. 2015
United Kingdom MFLUCC 16-0123 KX576655 KX576656 Hyde et al. 2016
Mycol Progress (2020) 19:1–14 7
71/1
/0.99
71/1
83/0.91
1
100/1
100/1
75/1
/1
78/1
100/0.99
82/1
100/1
94/1
100/1
88/
100/1
99/1
/0.99
91/1
96/1
100/1
100/1
100/0.98
100/1
60/
100/1
100/1
/0.94
100/1
1
87/1
92/1
88/
97/1
91/0.98
75/0.99
0.005
Plenodomus pimpinellae CBS 101637
Leptosphaeria sclerotioides CBS 144.84
Alternariaster centaureae-diffusae MFLUCC 14-0992
Plenodomus sp. (as Leptosphaeria sp. PHY-30)
Paraleptosphaeria praetermissa CBS 114591
Leptosphaeria errabunda CBS 617.75
Plenodomus lupini CBS 248.92
Heterospora dimorphospora CBS 165.78
Alternariaster helianthi CBS 199.86
Heterospora chenopodii CBS 115.96
Paraleptosphaeria dryadis CBS 643.86
Heterospora dimorphospora CBS 345.78
Alternariaster bidentis CBS 134021
Leptosphaeria doliolum CBS 155.94
Leptosphaeria slovacica CBS 389.80
Leptosphaeria doliolum MFLUCC 15-1875
Leptosphaeria doliolum CBS 541.66
Paraleptosphaeria sp. (as Leptosphaeria sp. PHY-54)
Subplenodomus violicola CBS 306.68
Paraleptosphaeria nitschkei MFLU 13-0644
Plenodomus wasabiae CBS 120120
Plenodomus wasabiae CBS 120119
Paraleptosphaeria macrospora CBS 114198
Plenodomus visci CBS 122783
Leptosphaeria pedicularis CBS 390.80
Leptosphaeria sydowii CBS 385.80
Leptosphaeria sydowii CBS 125976
Paraleptosphaeria polylepidis MA 57834
Leptosphaeria errabunda CBS 125978
Leptosphaeria doliolum CBS 505.75
Paraleptosphaeria dryadis CBS 743.86
Paraleptosphaeria nitschkei CBS 306.51
Plenodomus biglobosa CBS 298.36
Alternariaster helianthi CBS 327.69
Plenodomus biglobosa CBS 127249
Heterospora chenopodii CBS 448.68
Cucurbitaria berberidis CBS 394.84
Leptosphaeria pedicularis CBS 126582
Paraleptosphaeria rubi MFLUCC 14-0211
Paraleptosphaeria sp. (as Leptosphaeria sp. PHY-06)
Plenodomus salviae MFLUCC 13-0219
Alternariaster bidentis CBS 134185
Leptosphaeria sclerotioides CBS 148.84
Subplenodomus violicola CBS 100272
Paraleptosphaeria polylepidis APA 2999
Leptosphaeria slovacica CBS125975
Alternariaster centaureae-diffusae MFLUCC 15-0009
Fig. 1 Bayesian inference consensus tree inferred from a four-locus
dataset (SSU+ITS+LSU+TEF1) of the Leptosphaeriaceae clade.
Numbers above bold branches indicate bootstrap support values ≥70%
from maximum likelihood and posterior probabilities ≥0.95 from
Bayesian inference analyses. Cucurbitaria berberidis is used as an
outgroup
Tabl e 2 (continued)
Species Substrate/host Country Strain/voucher GenBank acc. no. References
ITS LSU
Pseudocoleophoma
typhicola
Submerged stems of
Typha latifolia
Pseudodictyosporium
elegans
––CBS 688.93 DQ018099 DQ018106 Tsui et al. 2006
Pseudodictyosporium
thailandica
Decaying bamboo
stem
Thailand MFLUCC 16-0029 KX259520 KX259522 Hyde et al. 2016
Pseudodictyosporium
wauense
––NBRC 30078 DQ018098 DQ018105 Tsui et al. 2006
Sajamaea mycophila Pseudothecia of
Paraleptosphaeria
polylepidis
Bolivia APA-2999 (DNA isolated
from conidial mass)
MK795715 MK795718 This study
Sajamaea mycophila Pseudothecia of
Paraleptosphaeria
polylepidis
Bolivia APA-2999 (DNA isolated
from conidiomata)
MK795716 MK795719 This study
APA A. N. Palabral-Aguilera; CBS CBS-KNAW Collections, Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; FMR Facultat de
Medicina i Ciencies de la Salut, Reus, Spain; HMAS Herbarium Mycologium Institute of Microbiology Chinese Academy of Sciences, Beijing, China;
KH K. Hirayama; KT K. Tanaka; KUMCC Culture collection of Kunming Institute of Botany, Kunming, China; MFLUCC Mae Fah Luang University
Culture Collection, Chiang Rai, Thailand; NBRC National Institute of Technology and Evaluation, Chiba, Japan; NTOU National Taiwan Ocean
University Culture Collection, Keelung, Taiwan; yone H. Yonezawa
8 Mycol Progress (2020) 19:1–14
closely located to the permanent monitoring plot “Cohiri,”
19K 512102, 8002927, elev. ca. 4425 m, parasitic on
Paraleptosphaeria polylepidis growing on Polylepis
tarapacana, 15 Oct. 2016, A.N. Palabral-Aguilera (APA-
2999), C. López & M.I. Gómez (LPB 0003512 –holotype,
KRAM F-59659 –isotype).
Discussion
Molecular phylogenetic analyses, based on the four-gene
dataset, placed Leptosphaeria polylepidis in a well-
supported clade consisting of members of the genus
Paraleptosphaeria, including the type species Pa.
nitschkei. The genus Paraleptosphaeria has been de-
scribed by de Gruyter et al. (2013)forPa. dryadis,
Pa. macrospora,Pa. nitschkei,Pa. orobanches,and
Pa. praetermissa that form a distinct monophyletic lin-
eage among Leptosphaeriaceae within Pleosporales.
Paraleptosphaeria is morphologically similar but clearly
divergent genetically from the Leptosphaeria lineage.
Separateness of Paraleptosphaeria from Leptosphaeria
has been confirmed by subsequent molecular studies
(Ariyawansa et al. 2015;Chenetal.2015; Liu et al.
2015; Wanasinghe et al. 2016; Tibpromma et al. 2017);
additionally, two novel species, Pa. padi and Pa. rubi,
have been described in the genus (Ariyawansa et al.
2015; Tibpromma et al. 2017) after its original estab-
lishment. Therefore, prior to this study, seven species
were accepted in Paraleptosphaeria.
The generic concept of Paraleptosphaeria includes species
having immersed, solitary or aggregated, thick-walled,
ostiolate and unilocular pseudothecia, bitunicate, 8-spored as-
ci, and fusiform, 5–8-septate, hyaline to yellow-brownish as-
cospores. The asexual state, when present, is coelomycetous
consisting of pycnidial, unilocular conidiomata, phialidic
conidiogeneous cells, and oblong or ellipsoidal, aseptate, hy-
aline conidia (de Gruyter et al. 2013). Leptosphaeria
polylepidis generally fits well to this concept, except that its
pseudothecia are superficial and developed on stroma.
However, this character may be variable in the genus, and,
for example, Liu et al. (2015) reported superficial
pseudothecia in their material of Paraleptosphaeria nitschkei.
Therefore, based on molecular evidence, Leptosphaeria
polylepidis is transferred to Paraleptosphaeria (as Pa.
polylepidis)inthisstudy.
98/1
100/1
90/1
/0.92
89/1
73/
/0.93
77/1
75/1
/0.92
99/1
94/1
82/
98/1
99/0.99
0.02
100/1
100/1
100/1
100/1
100/1
100/1
100/1
100/1
Pseudocoleophoma calamagrostidis KT 3284
Pseudodictyosporium wauense NBRC 30078
Pseudocoleophoma typhicola MFLUCC 16-0123
Dictyosporium thailandicum MFLUCC 13-0773
Pseudocoleophoma polygonicola KT 731
Periconia igniaria CBS 379.86
Dendryphiella vinosa NBRC 32669
Dictyocheirospora rotunda MFLUCC 17-1313
Neodendryphiella tarraconensis FMR 16234
Jalapriya pulchra MFLUCC 17-1683
Periconia igniaria CBS 845.96
Dictyosporium elegans NBRC 32502
Jalapriya inflata NTOU 3855
Neodendryphiella mali FMR 17003
Dendryphiella paravinosa CBS 14128
Sajamaea mycophila APA-2999 (Conidial mass)
Gregarithecium curvisporum KT 922
Dictyocheirospora bannica KH 332
Pseudodictyosporium elegans CBS 688.93
Neodendryphiella mali CBS 139.95
Jalapriya toruloides CBS 209.65
Sajamaea mycophila APA-2999 (Conidiomata)
Dendryphiella variabilis CBS 584.96
Dictyosporium bulbosum yone 221
Neodendryphiella michoacanensis FMR 16098
Cheirosporium triseriale HMAS 180703
Jalapriya pulchra MFLUCC 15-0348
Dictyocheirospora rotunda MFLUCC 14-0293b
Pseudodictyosporium thailandica MFLUCC 16-0029
Aquadictyospora lignicola MFLUCC 17-1318
Dictyocheirospora aquatica KUMCC 15-0305
Digitodesmium bambusicola CBS 110279
Fig. 2 Bayesian inference consensus tree inferred from two-locus dataset
(ITS+LSU) of the Dictyosporiaceae clade. Numbers above bold branches
indicate bootstrap support values ≥70% from maximum likelihood and
posterior probabilities ≥0.9 from Bayesian inference analyses. Periconia
igniaria is used as an outgroup
Mycol Progress (2020) 19:1–14 9
Other than genetically divergent, Paraleptosphaeria
may be ecologically different from Leptosphaeria by
having other host specialization. Five of the eight
Paraleptosphaeria species occur on Rosaceae, and three
species on other host families, Asteraceae,
Orobanchaceae, and Polygonaceae, while Leptosphaeria
species tend to inhabit hosts in very diverse families but
not Rosaceae (de Gruyter et al. 2013;Ariyawansaetal.
2015; Dayarathne et al. 2015; Liu et al. 2015). The
molecular phylogenetic analyses showed that
Paraleptosphaeria polylepidis was weakly supported as
sister species of Pa. dryadis supporting earlier finding
by Macía et al. (2005). These authors pointed out that
Pa. polylepidis and Pa. dryadis occur both at high ele-
vations. Their host plants, Polylepis tarapacana and
Dryas octopetala (Rosaceae), are adapted to grow in
cold ecosystems. According to our molecular analyses,
the branch leading to Pa. polylepidis is very long,
which may suggest an old evolutionary origin of this
lineage. It is, however, also possible that undiscovered
closely related Paraleptosphaeria species may still exist
in poorly studied South American countries. The weakly
supported sister-group relationship of Pa. polylepidis
and Pa. dryadis and the circumstance that their hosts
abc
d f g
eh
Fig. 3 Paraleptosphaeria polylepidis (KRAM F-59659). a,b,dHabit of
the crowded ascomata on twig of Polylepis tarapacana.cLongitudinal
section through ascomata and stroma. eParaplectenchymatous peridium
from longitudinal section. fHamathecium composed of thin interascal
filaments. gBitunicate asci with 5–8 ascospores inside (left mounted in
water, right mounted in KOH/IKI). hAscospores mounted in water. Scale
bars: a–d500 μm, e25 μm, f,h10 μm, g50 μm
10 Mycol Progress (2020) 19:1–14
are phylogenetically unrelated (Potter et al. 2007)sug-
gests that they evolved from different ancestral species.
The assignment of conidiomata found on the pseudothecia
of Paraleptosphaeria polylepidis to a new phylogenetic line-
age of the Dictyosporiaceae was unexpected as we earlier
hypothesized having encountered the asexual state of Pa.
polylepidis. The sequences obtained from the fungus formed
a strongly supported sister clade to members of the genus
Pseudocoleophoma but could not be classified in
Pseudocoleophoma on the basis of different morphological
characteristics. The genus Pseudocoleophoma has been de-
scribed for two species, Ps. calamagrostidis (generic type)
and Ps. polygonicola, that produced sexual states on the nat-
ural substrates and asexual states in cultures (Tanaka et al.
abc
d
f
g
i
h
e
Fig. 4 Sajamaea mycophila (KRAM F-59659). a–cHabit of pale brown
conidiomata growing on ascomata of Paraleptosphaeria polylepidis.d–e
Longitudinal section of uniloculate conidioma erumpent through the
outermost layer of host peridium (emounted in LPCB). fLongitudinal
section of multi-loculate conidioma showing layers of conidiogenous
cells (hyaline areas) and conidial masses (brown areas). g
Section through paraplectenchymatous peridium showing conidiogenous
cells mounted in LPCB. hConidiogenous cells and young conidia
mounted in LPCB. iConidia. Scale bars: a1000 μm, b–c500 μm, d–f
50 μm, g–i10 μm
Mycol Progress (2020) 19:1–14 11
2015). Two more species have been described later, Ps.
typhicola forming only asexual state on the natural substrate
(Hyde et al. 2016)andPs. bauhiniae producing both asexual
and sexual states on the natural substrate (Jayasiri et al. 2019).
While the fungus found on the pseudothecia of Pa. polylepidis
lacks a sexual state and forms uniloculate to multi-loculate
pycnidia and pale brown, broadly ellipsoidal conidia, the spe-
cies of Pseudocoleophoma usually produce a sexual state and
pycnidia of the asexual state are uniloculate and the cylindrical
conidia are hyaline. It also differs ecologically by parasitism
on another fungus, while all Pseudocoleophoma species are
saprobic on dead plant materials. Though we did not check
mycelial interactions in culture experiments, the fungus forms
pycnidia erumpent through the outermost layer of host perid-
ium and, therefore, is likely mycoparasitic (at least
necrotrophic as defined by Sun et al. 2019). Furthermore,
the molecular phylogenetic analyses indicated genetic dis-
tances between this fungus and Pseudocoleophoma compara-
ble to distances between other Dictyosporiaceae clades,
assigned to distinct genera (Boonmee et al. 2016). No
morphologicaly similar coelomycete genus was found in
Sutton (1980) and in the recent literature; therefore, a new
genus and species names, Sajamaea and S. mycophila, are
introduced for this fungus. The family Dictyosporiaceae con-
tains mostly asexual species, while sexual species are very
rare, being represented only by Dictyosporium meiosporum,
D. sexualis,Gregarithecium curvisporum,
Pseudocoleophoma calamagrostidis,P. polygonicola,and
P. bauhiniae (Tanaka et al. 2015; Boonmee et al. 2016;
Jayasiri et al. 2019). The asexual species are mostly hypho-
mycetous, often producing characteristic cheirosporous co-
nidia (Tanaka et al. 2015; Boonmee et al. 2016; Iturrieta-
González et al. 2018;Yangetal.2018), but Sajamaea and
Pseudocoleophoma have coelomycetous asexual states,
which supports their close phylogenetic relationship.
The parasitism of Sajamaea mycophila suggests that this
species may be potentially used as a biopesticide that could
prevent the development of Paraleptosphaeria polylepidis
and consequently protect Polylepis tarapacana woodlands.
Additional emphases are thus required to re-collect
S. mycophila and aim at culturing it in order to unravel its
mycoparasitic and biopesticidal potentials.
Acknowledgments We appreciate the support of Cecilia López and the
staff of the Sajama National Park for their collaboration in field work. We
are greatly indebted to all staff of the Herbario Nacional de Bolivia,
Instituto de Ecología, Universidad Mayor de San Andrés, La Paz, for
their generous long-term cooperation.
Funding information The study was funded by the International
Foundation for Science (IFS). Research on the mycoparasitic fungus
was financially supported by the National Science Centre (NCN) in
Poland (DEC-2013/11/D/NZ8/03274). MP, PRF and AF received addi-
tional support under statutory funds from the W. Szafer Institute of
Botany, Polish Academy of Sciences, Kraków, Poland.
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