Two novel species of Vagicola (Phaeosphaeriaceae) from Italy

Abstract

Phaeosphaeriaceae is a large and important family in the order Pleosporales, comprising economically important plant pathogens. Species may also be endophytes or saprobes on plant hosts. Two new species referable to Vagicola, Phaeosphaeriaceae are introduced in this paper based on analyses of LSU and ITS sequence data and their unique morphology. Most Phaeosphaeriaceae species grow on monocotyledons; Vagicola dactylidis and V. chlamydospora are also saprobic on grasses (Poaceae). Vagicola chlamydospora formed asexual structures in a culture. The new species are described and illustrated and compared with other taxa.

Full text

Submitted 24 October 2015, Accepted 19 November 2015, Published online 24 November 2015
Corresponding Author: Itthayakorn Promputtha  e-mail  ppam118@gmail.com 716
Two novel species of Vagicola (Phaeosphaeriaceae) from Italy
Jayasiri SC1, Wanasinghe DN1,2, Ariyawansa HA3, Jones EBG4, Kang JC5,
Promputtha I6, Bahkali AH4, Bhat J7,8, Camporesi E9 and Hyde KD1, 2, 4
1Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand
2World Agro forestry Centre East and Central Asia Office, 132 Lanhei Road, Kunming 650201, China
3Guizhou Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, 550006,
Guizhou, China
4Botany and Microbiology Department, College of Science, King Saud University, Riyadh, 1145, Saudi Arabia
5Engineering Research Center of Southwest Bio-Pharmaceutical Resources, Ministry of Education, Guizhou
University, Guiyang 550025, Guizhou Province, China
6Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand
7No. 128/1-J, Azad Housing Society, Curca, P.O. Goa Velha, 403108, India
8Department of Botany, Goa University, Goa, 403 206, India
9A.M.B. GruppoMicologicoForlivese “Antonio Cicognani”, Via Roma 18, Forlì, Italy; A.M.B. CircoloMicologico
“Giovanni Carini”, C.P. 314, Brescia, Italy; Società per gliStudiNaturalisticidella Romagna, C.P. 144, Bagnacavallo
(RA), Italy
Jayasiri SC, Wanasinghe DN, Ariyawansa HA, Jones EBG, Kang JC, Promputtha I, Bahkali AH,
Bhat J, Camporesi E, Hyde KD 2015  Two novel species of Vagicola (Phaeosphaeriaceae) from
Italy. Mycosphere 6(6), 716728, Doi 10.5943/mycosphere/6/6/7
Abstract
Phaeosphaeriaceae is a large and important family in the order Pleosporales, comprising
economically important plant pathogens. Species may also be endophytes or saprobes on plant
hosts. Two new species referable to Vagicola, Phaeosphaeriaceae are introduced in this paper
based on analyses of LSU and ITS sequence data and their unique morphology. Most
Phaeosphaeriaceae species grow on monocotyledons; Vagicola dactylidis and V. chlamydospora
are also saprobic on grasses (Poaceae). Vagicola chlamydospora formed asexual structures in a
culture. The new species are described and illustrated and compared with other taxa.
Key words  LSU  ITS  monocotyledons  multigene analyses  Poaceae
Introduction
Phaeosphaeriaceae is a large family in the order Pleosporales (Hyde et al. 2013,
Phookamsak et al. 2014). Members of this group grow mainly on monocotyledons, but some
species have also been reported on dicotyledons (Shoemaker and Babcock 1989, Schoch et al.
2006, Zhang et al. 2009, 2012, De Gruyter et al. 2010, Hyde et al. 2013, Wijayawardene et al.
2014). The family was introduced by Barr (1979) and recent studies have shown it to be a natural
group comprising 25 genera (Ariyawansa et al. 2015). Ariyawansa et al. (2015) and Phukhamsakda
et al. (2015) have provided the latest backbone trees for the family. The asexual morphs are
coelomycetous (Zhang et al. 2009, Phookamsak et al. 2014, Wijayawardene et al. 2014, Li et al.
2015). The family Phaeosphaeriaceae has a cosmopolitan distribution, and species are generally
Mycosphere 6 (6): 716–728 (2015) ISSN 2077 7019
www.mycosphere.org Article Mycosphere
Copyright © 2015 Online Edition
Doi 10.5943/mycosphere/6/6/7

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necrotrophic, plant pathogens or saprobes on a wide range of plants Shoemaker and Babcock 1989,
Carson 2005, Stukenbrock et al. 2006, Cannon and Kirk 2007.
The genus Phaeosphaeria was introduced by Miyake (1909). Miyake (1909) treated 114
species of Phaeosphaeria and accommodated them in six subgenera, viz. Ovispora, Fusispora,
Phaeosphaeria, Spathispora, Vagispora and Sicispora based on the differences in ascospore shape,
number of septa and the gelatinous sheaths on spores (Eriksson 1967, Shoemaker & Babcock
1989). The morphological characters of taxa in this genus are often ambiguous and can be confused
with other taxa in the Leptosphaeriaceae and Montagnulaceae, and with genera in the family itself
(Hyde et al. 2013, Phookamsak et al. 2014). Multigene phylogenetic analyses were carried out to
confirm the placement of this group by Zhang et al. (2009), Phookamsak et al. (2014) and
Ariyawansa et al. (2015). In this paper, we introduce two new species in Vagicola from Italy, which
were found on dead culms of Dactylis sp. (Poaceae). Combined analyses of LSU and ITS sequence
data using maximum-likelihood (ML) and maximum-parsimony (MP) clearly showed these species
grouped in Phaeosphaeriaceae with strong statistical support. In this paper, the two new species are
described and illustrated and compared with similar taxa.
Material and Methods
Collections, morphology and isolation
Specimens were collected in Italy by Erio Campesori. Study of gross morphology and
photomicrography were carried out under a stereomicroscope. Sections of ascoma were made free-
hand. Several specimens were used to observe the asci and ascospore characters and slides were
preserved in lactoglycerol. Micro-morphological characters were observed under a compound
microscope (Nikon Eclipse Ni), and measurements made using Tarosoft (R) Image Frame Work v.
0.9.7. Single spore isolation was carried out following the method of Chomnunti et al. (2014). Type
specimens of the new species are deposited in the Mae Fah Luang University Herbarium (MFLU),
Chiang Rai, Thailand, and ex-type cultures in Mae Fah Luang University Culture Collection
(MFLUCC) and Kunming Institute of Botany (KIB). Facesoffungi numbers and Index Fungorum
numbers are as outlined in Jayasiri et al. (2015) and Index Fungorum (2015).
Establishing the asexual morphs
Circular (0.5 cm) agar blocks from growing colony margins were cut and placed on fresh
Malt Extract Agar (MEA) plates as described in Phooksamak et al. (2015). Asexual structures
produced on Malt Extract Agar were observed after eight weeks of incubation, under light, at 20°
C.
DNA isolation, amplification and sequencing
Fungal isolates were grown on 2% MEA for 20 days at 16°C. Genomic DNA was extracted
from the growing mycelium using the Biospin Fungus Genomic DNA Extraction Kit-BSC14S1
(BioFlux®, P.R. China); following the instructions of the manufacturer (Hangzhou, P.R. China).
DNA sequence data was obtained from the internal transcribe spacer (ITS), large subunits of the
nuclear ribosomal RNA genes (LSU). Primer sets used for these genes were as follows: ITS:
ITS5/ITS4; LSU: LR0R/LR5 (Liu et al. 1999; Sung et al. 2007). The amplification was performed
following the instructions, and were set up for initial denaturation of 5 min at 95°C, followed by 35
cycles of 45 s at 94°C, 45 s at 52°C and 90 s at 72°C, and a final extension period of 10 min at
72°C. PCR-products were checked on 1% agarose electrophoresis gels stained with ethidium
bromide. Purification and sequencing of PCR products were done by Majorbio Co., China. DNA
sequence data were obtained from the large subunit rDNA (LSU) and internal transcribed spacers
will amplify by primer pairs ITS5 and ITS4 (White et al. 1990). Primer sequences and database are
available in GenBank. For Vagicola dactylidis single spore isolation was not successful. Therefore
fungal DNA was isolated directly from the ascomata.

718
Phylogenetic analysis
Sequences data were downloaded from GenBank to supplement the dataset (Table 1)
(Phookamsak et al. 2014, Ariyawansa et al. 2015). The represented sequences including those
newly obtained were aligned using with MAFFT v. 6.864b
(http://mafft.cbrc.jp/alignment/server/index.html) and improved manually where necessary using
Bioedit (Hall 1999). Didymella exigua was selected as outgroup taxon. The model of evolution was
carried out using MrModeltest 2.2 (Nylander 2004). Maximum likelihood analysis was performed
by using raxmlGUIv.0.9b2 (Silvestro and Michalak 2011). The search strategy was set to rapid
bootstrapping and the analysis carried out using the GTRGAMMAI model of nucleotide
substitution. The number of replicates was inferred using the stopping criterion (Pattengale et al.
2009). Maximum Likelihood bootstrap values equal or greater than 70% are given as the first set of
numbers above the nodes (Fig. 1). PAUPv4.0b10 was used to conduct the parsimony analysis to
obtain the phylogenetic trees. Trees were inferred using the heuristic search option with 1000
random sequence additions. Maxtrees were setup to 500 and branches of zero length were collapsed
and all multiple parsimonious trees were saved. Descriptive tree statistics for parsimony (Tree
Length [TL], Consistency Index [CI], Retention Index [RI], Relative Consistency Index [RC] and
Homoplasy Index [HI] were calculated for trees generated under different optimality criteria.
Kishino-Hasegawa tests (KHT) (Kishino and Hasegawa 1989) were performed in order to
determine whether trees were significantly different. Maximum-parsimony bootstrap values equal
or greater than 70% are given as the second set of numbers above the nodes (Fig. 1).
Results and Discussion
Molecular phylogeny
The combined LSU and ITS dataset comprising 57 strains of species of Phaeosphaeriaceae
were used to determine the generic placement of our two strains as Vagicola dactylidis and V.
chlamydospora. The phylogenetic trees obtained from Maximum Likelihood and Parsimony
analysis yielded trees with similar overall topology at subclass and family relationships, in
agreement with previous work based on Maximum Likelihood analysis (Zhang et al. 2012;
Phookamsak et al. 2013, 2014, Ariyawansa et al. 2014a, b, c, 2015, Wijayawardene et al. 2013,
Phukhamsakda et al. 2015). Individual LSU and ITS single gene trees were initially made and had a
similar topology (data not shown). Therefore the genes were combined. The maximum parsimony
dataset consists of 1360 characters with 987 characters as constant information, 113 characters as
variable characters are parsimony-uninformative, and 260 characters were count as parsimony-
informative character. The most parsimonious tree showed TL = 1873, CI = 0.343, RI = 0.613, RC
= 0.210, HI = 0.657 values. The best scoring tree is presented in Figure 1. The strains of Vagicola
dactylidis and V. chlamydospora clustered in the family Phaeosphaeriaceae. Vagicola dactylidis
and V. chlamydospora formed a sister clade with V. vagans (CBS 604.86) with 52% ML and 50%
MP support, but separate from other genera in the family. The new sequence data are deposited in
GenBank (Table 1).
Taxonomy
Vagicola K.W.T. Chethana and K.D. Hyde, in Ariyawansa et al. Fungal Diversity (2015)
= Phaeosphaeria subgen. Vagispora Shoemaker & Babcock, Can. J. Bot. 67: 15001599
(1989)
Type species: Vagicola vagans (Niessl) O. Eriksson, Chethana & K.D. Hyde, comb. nov.
Basionym: Pleospora vagans Niessl, Verh. Naturf. Ver. Briinn 14: 174. 1876
Phaeosphaeria vagans (Niessl) O.E. Erikss., Ark. Bot. 6: 430 (1967)
Vagicola chlamydospora Jayasiri, Camporesi & K.D. Hyde, sp. nov. Fig. 2
Index Fungorum Number: IF551683

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Facesoffungi Number: FoF 01323
Fig. 1 – RAxML Maximum Likelihood phylogenetic tree based on a combined LSU and ITS
sequence dataset. Bootstrap support values for Maximum Likelihood (ML) greater than 70% and
Maximum-Parsimony bootstrap values above 70% are given above and below the nodes
respectively. The tree is rooted to Didymella exigua (CBS 18355).

720
Table 1 Taxa used in the phylogenetic analysis and GenBank accession numbers (LSU and ITS)
and species. New sequences are in bold.
Taxon
Voucher/culture
numbers
GenBank accession numbers
LSU
ITS
Ampelomyces quisqualis
CBS 129.79
EU754128
HQ108038
Allophaeosphaeria cytisi
MFLUCC 15-0649
KT306950
KT306947
Allophaeosphaeria clematidis
MFLUCC 15-0652
KT306953
KT306949
Allophaeosphaeria dactylidis
MFLUCC 13-0618
KP744473
KP744432
Allophaeosphaeria subcylindrospora
MFLUCC 13-0380
KT314183
KT314184
Allophaeosphaeria muriformia
MFLUCC 13-0349
KP765681
KP765680
Chaetosphaeronema hispidulum
CBS 216.75
KF251652
KF251148
Coniothyrium dolichi
CBS 124140
GQ387611
JF740183
Coniothyrium glycines
CBS 124141
KF251714
KF251211
Dermatiopleospora mariae
MFLUCC 13-0612
KJ749653
KJ749654
Dermatiopleospora luzulae
MFLUCC 14-0932
KT306951

Didymella exigua
CBS 183.55
EU754155
GU237794
Diederichomyces cladoniicola
CBS 128026
JQ238628
KP170642
Diederichomyces caloplacae
CBS 129338
JQ238643
KP170639
Diederichomyces ficuzzae
CBS 128019
JQ238616
KP170647
Entodesmium rude
CBS 650.86
GU301812

Entodesmium artemisiae
MFLUCC 14-1156
KT315509
KT315508
Galliicola pseudophaeosphaeria
MFLUCC 14-0524
KT326693
KT326692
Leptospora rubella
CPC 11006
DQ195792
DQ195780
Loratospora aestuarii
JK 5535B
GU301838

Loratospora luzulae
MFLUCC 14 0826
KT328495
KT328497
Muriophaeosphaeria galatellae
MFLUCC 14-0614
KT438329
KT438333
Neosetophoma samarorum
CBS 139.96
GQ387579
KF251161
Neosetophoma samarorum
CBS 138.96
KF251664
KF251160
Neostagonospora caricis
CBS 135092
KF251667
KF251163
Neostagonospora elegiae
CBS 135101
KF251668
KF251164
Neosphaerellopsis thailandica
CPC 21659
KP170721
KP170652
Nodulosphaeria modesta
MFLUCC 11-0461
KM434285
KM434275
Nodulosphaeria senecionis
MFLUCC 15-1297
KT290257
KT290257
Ophiobolus cirsii
MFLUCC 13-0218
KM014662
KM014664
Ophiobolus erythrosporus
MFLUCC 12-2225
KM014665
KM491547
Ophiosphaerella aquatica
MFLUCC 14-0033
Ophiosphaerella agrostidis
MFLUCC 11-0152
KM434281
KM434271
Ophiosphaerella agrostidis
MFLUCC 12-0007
KM434282
KM434272
Paraphoma dioscoreae
CBS 135100
KF251671
KF251167
Paraphoma chrysanthemicola
CBS 172.70
KF251669
KF251165
Paraphoma radicina
CBS 111.79
KF251676
KF251172
Parastagonospora caricis
S6150/CBS135671
KF251680
KF251176
Parastagonospora nodorum
CBS 110109
KF251681
KF251177
Parastagonospora poae
CBS 135089
KF251682
KF251178
Phaeosphaeria papayae
CBS 135416
KF251690
KF251187
Phaeosphaeria alpina
CBS 456.84
KF251684
KF251181
Phaeosphaeria avenaria
DAOM 226215
AY544684


721
Taxon
Voucher/culture
numbers
GenBank accession numbers
LSU
ITS
Phaeosphaeria eustoma
CBS 573.86
DQ678063

Phaeosphaeria chiangraina
MFLUCC 13-0231
KM434280
KM434270
Phaeosphaeria musae
MFLUCC 11-0133
KM434277

Phaeosphaeria musae
CBS 120026
GU301862
DQ885894
Phaeosphaeria nigrans
CBS 307.79
KF251687

Phaeosphaeria oryzae
CBS 110110
KF251689
KF251186
Phaeosphaeria oryzae
MFLUCC 11-0170
KM434279
KM434269
Phaeosphaeria thysanolaenicola
MFLUCC 10-0563
KM434276
KM434266
Phaeosphaeria typharum
CBS 296.54
KF251695
KF251192
Phaeosphaeria vagans
CBS 604.86
KF251696
KF251193
Phaeosphaeriopsis musae
CBS 120026
GU301862
DQ885894
Phaeosphaeriopsis dracaenicola
MFLUCC 11-0157
KM434283
KM434273
Phaeosphaeriopsis glaucopunctata
MFLUCC 13-0265
KJ522477
KJ522473
Phaeosphaeriopsis triseptata
MFLUCC 13-0271
KJ522479
KJ522475
Populocrescentia forlicesenensis
MFLUCC 15-0651
KT306952
KT306948
Sclerostagonospora cycadis
CBS 123538
FJ372410
FJ372393
Scolicosporium minkeviciusii
MFLUCC 12-0089
KF366382

Septoriella phragmitis
CBS 140065
KR873279
KR873251
Septoriella oudemansii
CBS 138012
KJ869224
KR873250
Septoriella poae
CBS 136766
KJ869169
KJ869111
Septoriella leuchtmannii
CBS 459.84
KF251691
KF251188
Setophoma achromolaena
CBS 135105T/CPC
18553
KF251747
KF251244
Setophoma sacchari
CBS 333.39
KF251748
KF251245
Setophoma sacchari
MFLUCC11-0154
KJ476146
KJ476144
Setophoma terrestris
CBS 335.29
KF251749
KF251246
Sulcispora pleospora
MFLUCC 14-0995
Stagonospora neglecta
CBS 343.86
EU754218
AJ496630
Stagonospora foliicola
CBS 110111
KF251759
KF251256
Vagicola chlamydospora
MFLUCC 15-0177
KU163654
KU163658
Vagicola dactylidis
MFLU 15-2720
KU163656
KU163657
Vagicola vagans
CBS 604.86
KF251696
KF251193
Vrystaatia aloeicola
CBS 135107
KF251781
KF251278
Wojnowiciella viburni
MFLUCC 12-0733/ICMP
19778
KC594287
KC594286
Wojnowicia dactylisicola
MFLUCC 13-0738
KP684147
KP744469
Wojnowicia dactylisii
MFLUCC 13-0735
KP684149
KP744470
Wojnowicia lonicerae
MFLUCC 13-0737
KP684151
KP744471
Wojnowiciella eucalypti
CBS 139904
KR476774
KR476741
Xenophoma puncteliae
JDL-2012a/CBS 128022
JQ238619
JQ238617
Xenoseptoria neosaccardoi
CBS 128665
KF251784
KF251281
CBS Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands; CPC Working collection of
Pedro Crous housed at CBS; DAOM Plant Research Institute, Department of Agriculture
(Mycology), Ottawa, Canada; ICMP International Collection of Microorganisms from Plants, New
Zealand; JK: J. Kohlmeyer; MFLUCC Mae Fah Luang University Culture Collection, Chiang Rai,
Thailand.

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Fig. 2 – Vagicola chlamydospora (MFLU 15-1399, holotype) a-c Ascomata developing on surface
of host. d Papilla. e Section through the ascoma. f Peridium. g Pseudoparaphyses. h-j Asci. k-n
Ascospores. Scale bars: d = 30 µm, e = 50 µm, f, h-j = 10 µm, g, k-n = 5 µm.
Holotype  MFLU 15-1399
Etymology  With reference to chlamydospores-like asexual morph formed in culture
Saprobic on Dactylis sp. Sexual morph: Ascomata 121  208 
scattered or sometimes clustered, immersed, visible as raised, black dots on the host surface, uni- to
bi-loculate, subglobose, brown to dark brown, ostiole central, with a minute papilla. Peridium 21
27   -walled, of equal thickness, composed of 25 layers of brown to dark brown,
pseudoparenchymatous cells, arranged in textura angularis to inner layer composed of brown cells
of textura prismatica. Hamathecium composed of numerous, 1.72.3    ,
cellular pseudoparaphyses, with distinct septa, slightly constricted at the septa, embedded in a

723
Fig. 3 – Vagicola chlamydospora asexual morph in culture (MFLUCC 15-0177, ex-type culture).
a Fungal mycelium in MEA from above. b Fungal mycelium in MEA from below. c Close up view
of fruiting body in culture. d-h Asexual spores formation structures. Scale bars: d-h = 10 µm.
gelatinous matrix. Asci 5457 × 9     × 9.6   = 20), 8-spored, bitunicate,
fissitunicate, broadly cylindrical or cylindric-clavate, sessile to subsessile, apically rounded with an
ocular chamber. Ascospores 2132 × 3.7         = 30), overlapping 13-
seriate, phragmosporous, narrowly fusiform, with obtuse ends, with guttules, hyaline to yellowish-
brown, 9-septate, slightly curved, constricted at the septa, smooth-walled, lacking a sheath or
appendages. Asexual morph: Colonies on MEA effuse, white to pale yellow. Mycelium 23 
wide, prostrate, composed of septate, branched, smooth, hyaline, hyphae. Conidiophores micro- to
macronematous, erect, flexuous, smooth, composed of beaded, variedly-sized cells. Conidiogenous
cells 59 × 36 holoblastic, globose to subglobose, smooth, subhyaline. Conidia 912 × 1013

724
   10 × 11   = 20), globose, solitary, thick-walled, chlamydospore-like, with dense
cytoplasm (Fig 3).

725
Fig. 4 – Vagicola dactylidis (holotype). a, b Ascomata developing on surface of host. c Peridium. d
Section through ascoma e Papilla. f Pseudoparaphyses. g-j Asci. k-o Ascospores. Scale bars: a =
500 µm, b = 200 µm, c = 10 µm, d = 50 µm, e = 20 µm, g-j = 20 µm, g, f, k-o = 5 µm.
Material examined  ITALY, Province of Forlì-Cesena, near Poderone - Corniolo, dead
stem of Dactylis sp. (Poaceae), 21 October 2014, Erio Camporesi IT 2188 (MFLU 15-1399,
holotype), (isotype in KUN), ex-type culture, MFLUCC 15-0177, Genbank accession numbers:
LSU- KU163654, ITS-KU163658, SSU-KU163655.
Culture characters  Ascospores germinating on MEA within 36 h. Colonies growing on
MEA, reaching 2 cm diam. in 1 week at 16°C. Mycelium superficial, felty, gummy, edge undulate,
from above white, reverse yellow colour.
Notes  Vagicola chlamydospora resembles to Vagicola vagans in having similar ascomata,
as black coloured dots of ostiole visible with immersed ascomata when viewed on the host surface,
textura angularis to textura prismatica brown colour cells peridium. and broad cellular
pseudoparaphyses but differs in processing longitudinal septa and narrow ascospores. The spores in
Vagicola chlamydospora are similar to Nodulosphaeria, but differ in the morphology of ascomata
and phylogenetic analyses. In this study we observed chlamydospore-like asexual morph in culture
(Fig. 3).
Vagicola dactylidis Wanasinghe, Jayasiri, Camporesi & K.D. Hyde, sp. nov. Fig. 4
Index Fungorum Number: IF551684
Facesoffungi Number: FoF 01324
Holotype  MFLU 15-2720
Etymology  With reference to the host occurrence
Saprobic on dead stem of Dactylis sp. Sexual morph: Ascomata  

 solitary, scattered, superficial, globose to subglobose, dark
brown to black, coriaceous, ostiolate. Ostiole     
  
µm, n = 5), papillate, black, smooth, filled with dark brown cells. Peridium 912 µm wide at the
base, 1216 µm wide in sides, comprising 34 layers, comprising blackish to dark brown, thick-
walled cells of textura angularis. Hamathecium comprising numerous, 22.5 µm (n = 30) wide,
filamentous, branched, septate, pseudoparaphyses. Asci 

= 40), 8-spored, bitunicate, fissitunicate, broadly-clavate, with a short, orbicular pedicel, rounded at
apex, with minute ocular chamber. Ascospores     
        
obliquely bi-seriate, initially hyaline, becoming yellowish brown at maturity, broadly fusiform, 5-
trans-septate, with a longitudinal septa in between second and third trans-septate, constricted at the
central septa, weakly constricted at the other septa, with conical and narrowly rounded ends,
lacking a mucilaginous sheath. Asexual morph: Undetermined.
Material examined  ITALY, Province of Arezzo, Bagno di Cetica, dead stem of Dactylis
sp. (Poaceae), 8 October 2012, Erio Camporesi. IT 799 (MFLU 15-2720, holotype), (isotype in
KUN), GenBank accession numbers: LSU  KU163656, ITS  KU163657.
Notes  Vagicola dactylidis is more similar to the type species, V. vagans, than V.
chlamydospora, in having ascospores with vertical septa. Dactylis is the host plant of both Vagicola
chlamydospora and dactylidis. Two other species associated with Dactylis in Phaeosphaeriaceae
are Phaeosphaeria huronensis (Shoemaker & Babcock 1989) and Ophiosphaerella herpotricha
(Phookamsak et al. 2014), however, they share no morphological characters.
Acknowledgements
This work was supported by the International Research Group Program (IRG-14-27),
Deanship of Scientific Research, King Saud University, Saudi Arabia. K.D. Hyde thanks the
Chinese Academy of Sciences, [project number 2013T2S003], for the award of Visiting
Professorship for Senior International Scientists at Kunming Institute of Botany. MFLU [grant
number 56101020032] is thanked for supporting studies on Dothideomycetes. We are grateful to

726
the Mushroom Research Foundation, Chiang Rai, Thailand. Subashini C. Jayasiri is grateful to Mr.
and (Late) Mrs. Jayasiri and S.P.R.D. Lasantha for their valuable suggestions.
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