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ORIGINAL ARTICLE
Tristratiperidium microsporum gen. et sp. nov. (Xylariales) on dead
leaves of Arundo plinii
D. A. Daranagama
1,2
& E. Camporesi
4,5,6
& X. Z. Liu
1
& D. J. Bhat
8
&
S. Chamyuang
2
& A. H. Bahkali
7
& M. Stadler
3,4
& K. D. Hyde
2,7
Received: 7 September 2015 /Revised: 19 November 2015 /Accepted: 10 December 2015
#
German Mycological Society and Springer-Verlag Berlin Heidelberg 2015
Abstract Trist ratiperidium microsporum gen. et sp. nov.
(Xylariales) is introduced to accommodate a taxon isolated
from dead leaves of Arundo plinii, collected in Italy. The sexual
and asexual morphs are described and illustrated and compared
with similar taxa. Phylogenetic analysis of combined Internal
T ranscribed Spacer (ITS) and Large Subunit (LSU) rRNA se-
quence data show the relationships of T. microsporum with
other genera in the Xylariales. Tristratiperidium micro sporum
clusters with the hyphomycete Subramaniomyces
fusisaprophyticus in the order Xylariales, in the clade compris-
ing Apiosporaceae, Hyponectriaceae, Melogrammataceae, and
Pseudomassariaceae. The phylogenetic relationships of
T. micr osporum and allied fungi are discussed and its placement
in the Xylariales genera incertae sedis is suggested.
Keywords Ascomycetes
.
Phylogeny
.
Sordariomycetes
.
Taxonomy
.
Xylariales
Introduction
The Xylariales is a large order of perithecial ascomycetes with
unitunicate asci, accommodated in the subclass
Xylariomycetidae (Zhang et al. 2006; Maharachchikumbura
et al. 2015). The families and genera of the Xylariales have
been traditionally segregated based on morphology. This clas-
sification has been artificial as well as problematic, as the
relative importance of different characters was unclear. The
varied morphological features of members of Xylariales led
to different classification systems. Therefore, several different
classification systems, predominantly based on morphology
were introduced by different authors (Munk 1953; Müller
and von Arx 1973;Barr1990;Hawksworthetal.1995;
Eriksson et al. 2003). Eriksson et al. (2003) and Smith et al.
(2003) used molecular data to infer the phylogenetic relation-
ships within the order. Smith et al. (2003) analyzed combined
datasets of 28S and 18S rDNA sequence data to support the
Xylariales as a monophyletic order, in which
Amphisphaeriac eae, Apiospora ceae, Clypeosph aeriace ae,
Diatrypaceae, Graphostromataceae, Hyponectriaceae, and
Xylariaceae were recognized. However, due to the lack of
reliable strains and sequence data used in this study, they
could not elucidate the famili al relationships within the
Xylariales with certainty (Eriksson et al. 2003; Smith et al.
2003;Zhangetal.2006;Tangetal.2009; Triebel et al.
2005). However, members of the Xylariales generally have
stromata, but sometimes stromata can be absent or highly
Section Editor: Gerhard Rambold
* X. Z. Liu
liuxz@im.ac.cn
1
State Key Laboratory of Mycology, Institute of Microbiology,
Chinese Academy of Sciences, No 3 1st West Beichen Road,
Chaoyang District, Beijing 100101, People’s Republic of China
2
Center of Excellence in Fungal Research, Mae Fah Luang University,
Chiang Rai 57100, Thailand
3
Helmholtz-Zentrum für Infektionsforschung GmbH, Department of
Microbial Drugs, Inhoffenstrasse 7, 38124 Braunschweig, Germany
4
A.M.B. Gruppo Micologico Forlivese BAntonio Cicognani^,Via
Roma 18, Forlì, Italy
5
A.M.B. Circolo Micologico BGiovanni Carini^,C.P.
314 Brescia, Italy
6
Società per gli Studi Naturalistici della Romagna, C.P.
144 Bagnacavallo, RA, Italy
7
Department of Botany and Microbiology, King Saudi University,
Riyadh, Saudi Arabia
8
Department of Botany, Goa University, Goa 403206, India
Mycol Progress (2016) 15:8
DOI 10.1007/s1 1557-015-1151-y
reduced, thick-walled perithecial ascomata, usually 8-spored
unitunicate asci, with a J+, or J– apical apparatus, and
pigmented ascospores with germ slits or germ pores and mu-
cilagino us sheath or appendages ( Barr 1990; Hawksworth
et al. 1995; Smith et al. 2003).
Zhang et al. (2006) and Maharachchikumbura et al.
(2015 ) provided an updated outline for Sordariomycetes
based on a multigene analysis [28S, 18S, translation e lon-
gation factor (TEF) and RNA polymerase II (RPB2) se-
quence data]. Maharachchikumbura et al. (2015)included
Amphisphaeriaceae, Apiosporaceae, Cainiaceae,
Clypeosphaeriaceae, Coniocessiaceae, Diatrypaceae,
Graphostromataceae, Hyponectriaceae, Melogrammataceae,
Vialaeaceae, and Xylariaceae in Xylariales,
while Hernandez– Restrepo et al. (2015)included
Amphisphaeriac eae, Apiospora ceae, Clypeosph aeriace ae,
Diatrypaceae, Hyponectriaceae, Microdochiaceae, and
Xylariaceae. Senanayake et al. (2015) revised the group using
combined ITS and LSU analysis in a move towards redefining
Xylariomycetidae and resurrected the order Amphisphariales
and provided an updated classification for Xylariales. Accord-
ing to Senanayake et al. (2015), the order Xylariales com-
prises the families recognized by Maharachchikumbura et al.
(2015), plus Lopadostomaceae and Pseudomassariaceae.
Amphisphaeriaceae and Clypeosphaeriaceae were removed
from Xylariales and placed in Amphisphaeriales
(Senanayake et al. 2015).
In this study, we introduce a new genus Tristratiperidium in
the order Xylariales. Morphological characters as well as phy-
logenetic analysis indicate that T ristratiperidium belongs in
Xylariales, although it is not linked to any of the existing
families.
Materials and methods
Sample collection and specimen examination
A specimen was collected in Italy in October 2014, and mac-
roscopic and microscopic characters were recorded. A Motic
SMZ-168 dissecting microscope (Ted Pilla. Inc, USA) was
used to observe the structures of the ascomata. Asci and asco-
spores were examined using a Nikon ECLIPSE 80i compound
microscope (Nikon, Tokyo, Japan). Melzer’s reagent and
Lugol solution were applied to observe if the ascus apical
apparatus stained blue. Measurements of stromata (n = 10),
ascomata (n =10),asci (n = 20), and ascospores (n = 40) were
made from material mounted in water and the mean values
were calculated. Photomicrography was carried out using a
Canon 450D digital camera (Canon Inc., Tokyo, Japan) fitted
to the microscope. Measurements were made with the
Tarosoft (R) Image Frame Work program and images used
for figures were processed with Adobe Photoshop CS3
Extended version 10.0 software (Adobe Systems Inc). Herbar-
ium material is deposited in the herbaria of Mae Fah Luang
University (MFLU), Thailand and Kunming Institute of Bot-
any (KUN), China and cultures are deposited at Mae Fah
Luang University Culture Collection (MFLUCC), Thailand
and the Kunming Institute of Botany Culture Collection
(KIBCC), and China General Microbiological Culture Collec-
tion Center (CGMCC) China. Faces of fungi numbers and
Index Fungorum numbers are as explained in Jayasiri et al.
(2015) and Index Fungorum (2015).
Description of cultures and asexual morph
Pure cultures were obtained from single spores, following the
method detailed by Chomnunti et al. (2014). The cultures
were grown in malt and yeast extract agar (Malt extract 6 g/
L, yeast extract 0.6 g/L, dextrose 4 g/l) and incubated at 25–
28 °C for 2–4 days. After 2–4 days, hyphal tips were cut and
transferred to fresh Difco Oatmeal Agar (OA). The cultures
were incubated at 25–28 °C for 1 month. After 2–3weeks,
cultures on OA were checked for asexual structures.
Conidiogenous structures (conidiophores, conidiogenous
cells, and conidia) were observed and measured by phase
contrast microscopy under 400–1000× optical magnification.
DNA isolation, PCR, and sequencing
DNA was extracted from isolates grown on malt and yeast
extract agar media overlaid with sterilized cellophane for
5 days at 25 °C. DNA isolation and purification was carried
out according to Daranagama et al. (2015). For amplification
of ITS and LSU loci the primers and PCR protocols detailed in
Daranagama et al. (2015) were used. The DNA fragments
were amplified using an automated thermal cycler (DongShen
EDC-810- Eastwin, China). A total volume of 50 μl
[10 × PCR buffer, 0.25 mM dNTP, 0.4 μMofeachprimer;
1.5 mM MgCl
2
, 0.8 units Taq Polymerase and 10 ng template
DNA (1:10 diluted)], was used for PCR with adjustments of
component volumes and concentration when needed. Th e
PCR products were visualized on 1 % agarose gels stained
with Goldview (Geneshun Biotech, China) with D2000
DNA ladder (Realtimes Biotech, Beijing, China). All PCR
products were purified according to the company protocols
and DNA sequencing was performed using the same primers
in an Applied Biosystem 3730 DNA analyzer at Omega Ge-
netics Company, Beijing, China. The sequences derived from
this study were deposited in GenBank (Table 1).
Sequence alignment and phylogenetic analysis
To reveal the phylogenetic position of Tristratiperidium,52
ingroup t axa from representative xylari alean species were
downloaded fr om GenBank and include d in the analysis
8 Page 2 of 8 Mycol Progress (2016) 15:8
Table 1 GenBank Accession
numbers of the strains used in this
study
Species name Strain number GenBank Accession numbers
ITS LSU
Amphibambusa bambusicola MFLUCC 11–0617 KP744474 KP744474
Annulohypoxylon nitens MFLUCC 12-0823 KJ934991 KJ934992
Apiospora bambusae ICMP 6889 – DQ368630
Apiospora setosa ICMP 4207 – DQ368631
Arecophila bambusae HKUCC 4794 – AF452038
Arthrinium hyphopodii MFLUCC 15–0003 KR069110 KR069111
Arthrinium phaeospermum HKUCC 3395 – AY0838 32
Arthrinium subglobosa MFLUCC 11–0397 KR069112 KR069113
Astrocystis concavispora MFLUCC 14–0174 KP297404 KP340545
Atrotorquata spartii MFLU 14–0738 – KP325443
Biscogniauxia marginata MFLUCC 12-0740 KJ958407 KJ958408
Cainia anthoxanthis MFLUCC 15–0539 KR092787 KR092777
Cainia graminis MFLUCC 15–0540 KR092793 KR092781
Cainia graminis CBS 136.62 AF431949 AF431949
Camillea obularia ATCC28093 AF201714 –
Collodiscula japonica CBS 124266 JF440974 JF440974
Coniocessia maxima CBS 593.74 GU553332 GU553344
Coniocessia nodulisporioides CBS 281.77 – GU553352
Creosphaeria sassafras CM AT-018 – DQ840056
Daldinia concentrica CBS113277 AY616683 KT281895
Diatrype disciformis MFLUCC 15–0538 KR092795 KR092784
Diatrype palmicola MFLUCC 11–0018 KP744439 KP744481
Diatrype whitmanensis ATCC MYA- 4417 F J746 65 6 –
Eutypa flavovirens MFLUCC 13–0625 KR092798 KR092774
Eutypa lata CBS 208.87 = AFTOL-ID 929 DQ006927 DQ836903
Graphostroma platystoma AFTOL-ID 1249 DQ836906 DQ836906
Hyponectria buxi UME 31430 – AY0838 34
Hypoxylon fragiforme MUCL 51264 KM186294 KM186295
Iodosphaeria tongrenensis GZUH0109 = FJS8 KR095282 KR095283
Lopadostoma americanum LG8 KC774568 KC774568
Lopadostoma dryophilum LG21 KC774570 KC774570
Lopadostoma fagi
LF1 KC774575 KC774575
Lopadostoma quercicola LG27 KC774610 KC774610
Lopadostoma turgidum LT2 KC774618 KC774618
Lunatiannulus irregularis MFLUCC 14–0014 KP297398 KP340540
Melogramma campylosporum MBU JF440978 JF440978
Ophiodiaporthe cyatheae YMJ 1364 JX570889 JX570891
Plagiostoma aesculi AFTOL-ID 1238 – DQ836905
Poronia pileiformis WSP 88113001 GU324760 –
Pseudomassaria chondrospora PC1 JF440982 JF440982
Pseudomassaria chondrospora MFLUCC 15–0545 KR092790 KR092779
Pseudomassaria sepincoliformis CBS 129022 JF440984 JF440984
Repetophragma inflatum NN 42958 – DQ408576
Sarcoxylon compunctum CBS359.61 KT281903 KT281898
Seynesia erumpens SMH 1291 AF279410 AF279410
Sporidesmium knawiae CPC 15467 FJ349609 FJ349610
Subramaniomyces fusisaprophyticus CBS 418.95 EU040241 EU040241
Tristratiperidium microsporum MFLUCC 15-0413 KT696538 KT696539
Vialaea mangiferae MFLUCC 12–0808 KF724974 KF724975
Vialaea minutella BRIP 56959 – KC181924
Xylaria hypoxylon CBS122620 AM993141 KM186301
Xylaria obovata MFLUCC 13–0115 KR049088 KR049089
Mycol Progress (2016) 15:8 Page 3 of 8 8
(Table 1). Plagiostoma aesculi (Fuckel) Sogonov (AFTOL-ID
1238) and Ophiodiaporthe cyatheae Y.M. Ju, et al. (YMJ
1364), belonging to Diaporthales were used as outgroup. Phy-
logenetic analysis was performed using a combined ITS–28S
sequence data matrix.
Sequence data were initially aligned with MUSCLE v.3.6
(Edgar 2004) and Bioedit 7.1.3.0 (Hall 1999) and optimized
with Clustal X v1.83 (Thompson et al. 1997) and manually
aligned where necessary. All characters were assessed to be
unordered and equally weighed. Gaps were treated as missing
data. Phylogenetic analyses were performed using RAxML
v7.0.3 (Stamatakis and Alachiotis 2010)asimplementedin
RAxML GUI 0.95 (Silvestro and Michalak 2012). The search
strategy was set to rapid bootstrapping and the analysis carried
out using the GTR model of nucleotide substitution. The model
of evolution was estimated by using MrModeltest 2.2
(Nylander 2004). The bootstrap analysis for each ML tree
was performed with 1000 fast bootstrap replicates with the
same parameter settings using the GTR substitution model se-
lected by MrModeltest 2.2. Model parameters were selected
independently for the different gene regions under the Akaike
Information Criterion (AIC) implemented in PAUP v.4. The
resulting trees were viewed using the Tree View application
(Page 1996). The alignment was submitted to TreeBASE
(http://purl.org/phylo/treebase/phylows/study/TB2:S18169?x-
accesscode=4f56888bab25c34e125d7ef290f32081&format=
html).
Results
Molecular phylogeny
The familial placements within Xylariales are similar to those
in Senanayake et al. (2015). The order Xylariales comprises
15 clades (A–O) which represent the families Apiosporaceae,
Cainiaceae, Coniocessiaceae, Diatrypaceae, Hyponectriaceae,
Iodosphaeriaceae, Lopadostomaceae, Melogrammataceae,
Pseudomassariaceae, Vialaeaceae, hypoxyloid and xylarioid
Xylariaceae, and three unnamed clades: clade B with
Graphostroma platystoma (Schwein.) Piroz., clade L includ-
ing Subramaniomyces fusisaprophyticus (Matsush.) P.M.
Kirk and Tristratiperidium microsporum, and clade O com-
prising Repetophragma inflatum (Berk. & Ravenel) W.P. Wu
and Sporidesmium knawiae Crous (Fig. 1).
Tristratiperidium clustered with Subrama niomyces
fusisaprophyticus as a distinct clade in Xylariales with 86 %
bootstrap support, but did not cluster in any of the existing
families. Thus, we introduce the new genus in Xylariales gen-
era incertae sedis. Tristratiperidium microsporum together
with Subramaniomyces fusisaprophyticus form a sister clade
to the lineage comprising Apiosporaceae, Hyponectriaceae,
Melogrammataceae, and Pseudomassariaceae.
Taxonomy
Tristratiperidium Daranagama, Camporesi & K.D. Hyde,
gen. nov.
Index Fungorum number: IF 551386, Facesoffungi num-
ber: FoF 00919
Etymology: Refers to the three-layered peridiu m of the
ascomata; tri [= three] + stratum [= layer] + peridium.
Saprobic on dead leaves of grasses. Sexual morph:
Ascomata immersed to partially erumpent, coriaceous, visible
as shiny black dots, globose, solitary, with papillate ostiole.
Papilla dark brown, central, projecting above the surface.
Peridium comprising three layers, with outer layer comprising
thick-walled, dark brown cells of textura angularis,central
layer comprising thick-walled, hyaline cells of textura
intricata, and innermost layer comprising thin-walled,
hyaline-light brown cells of textura prismatica. Hamathecium
comprising numerous, 2 μm wide, filamentous paraphyses.
Asci 8-spored, unitunicate, cylindrical, short pedicellate, with
J- apical apparatus. Ascospores uniseriate, dark brown, unicel-
lular, ellipsoid-subglobose, with a straight, full length germ
slit. Asexual morph: Hyphomycetous, sporulating regions
produced in 1-week-old culture with ab undant conidia.
Conidiophores simple, brown, unbranched, erect, straight, or
slightly flexuous. Conidiogenous cells phialidic, terminal,
percurrently proliferating. Conidia phragmosporous, hyaline,
fusiform, bearing two back-curved, short, terminal setulae.
Type species: T ristratiperidium micr osporum Daranagama,
Camporesi & K.D. Hyde
Notes: The new genus Tristratiperidium was isolated from
fallen dead leaves of Arundo plinii. According to the phylo-
genetic analysis it clustered in Xylariales. This genus superfi-
cially resembles certain xylariaceous fungi, with partially
erumpent, black, ostiolate ascomata. The three-layered perid-
ium with different cell structures and obovoid, light brown
ascospores as well as the asexual morph of Tristratiperidium,
which is considered the most salient feature, discriminates it
from other known taxa (see discussion).
Tristratiperidium microsporum Daranagama, Camporesi
&K.D.Hyde,sp. nov.
Index Fungorum number: IF551387, Facesoffungi num-
ber: FoF 00920 Figs. 2 and 3
Etymology: Refers to the small ascospores.
Habitat: On dead leaves of Arundo plinii L. Sexual
morph: Ascom ata (120–)185 –2 35(− 257) × (150–) 161–
190(−211) μ
m (=215 × 185 μm, n =10),immersedtopartially
erumpent, coriaceous, surface convex, smooth, visible as
shiny black dots, surrounded by white fungal hyphae, glo-
bose, solitary, scattered, ostiolate, papillate. Papilla (7–)10–
15(−17) μm wide (=12.5 μm, n = 10), central, dark brown,
projecting above the surface. Peridium (15–)20–27(−29) μm
wide (=22.5 μm, n = 10), clearly-defined, comprising three
main layers with different cellular arrangements, outer layer
8 Page 4 of 8 Mycol Progress (2016) 15:8
comprisi ng thick-w alled, dark brown cells of textura
angularis, central layer comprising thick-walled, hyaline cells
of textura intricata, and innermost layer comprising thin-
walled, hyaline-light brown cells of textura prismatica.
Hamathecium comprising 2 μm wide, numerous, filamentous
paraphyses. Asci (65–)72–106(−119) × (5.7–)6.6–7.8(−8.2)
μm (=96 × 7.4 μm, n = 20), 8-spored, unitunicate, cylindrical,
short pedicellate, apex slightly thickened, with J- apical appa-
ratus. Ascospores (8–)8.4–10.5(−11) × (4.8–)5.1–6.5(−6.7)
μm (=9.7 × 5.6 μm, n = 20), uniseriate, dark brown, unicellu-
lar, ellipsoidal-subglobose, some with broad ends, smooth-
walled, with a straight, full-length germ slit. Asexual morph:
Hyphomycetous, sporulating regions produced in 1 week old
culture with abundant conidia. Conidiophores
macronematous, (42–)50–80(− 91) × (5–)5.5–8(− 8.4) μm
(=73 × 7.6 μm, n = 20), simple, brown, unbranched, thick-
walled, erect, straight or slightly flexuous. Conidiogenous
cells terminal, phialidic, percurrently proliferating, less than
Fig. 1 Phylogram inferred from likelihood analysis of members of
Xylariales using combined ITS and 28S sequence data. Strain/culture
numbers are given following the taxon names. The new sequences
generated in this study are in blue. The bootstrap support values from
likelihood analysis >50 % from 1000 RAxML replicates are shown above
or below the branches. The tree is rooted with Plagiostoma aesculi
(AFTOL-ID 1238) and Ophiodiaporthe cyatheae (YMJ 1364)
Mycol Progress (2016) 15:8 Page 5 of 8 8
5 μm long, with minute collarette. Conidia (8–)9.2–
13.5(−15) × (3–)4–5(−5.4) μm (=10.8 × 4.6 μm, n =20), hya-
line, fusiform, slightly curved, 1– to 2-septate, gutullate, bear-
ing two back-curved, short, polar setulae.
Culture characteristics: Colonies on Difco OA reaching
the edge of 9 cm Petri-dish in 4 weeks at 25–27 °C, at first
whitish, felty, azonate, with diffuse margins, developing spor-
ulating regions as light grey spots in the centre of the culture;
reverse turn ing light grey after 2 we eks. Producing dense
masses of conidiophores.
Specimen examined: Italy, Pr ovince of Forlì-Cesena,
Porcia–Pre dappio, on de ad leaves of Arundo plinii L
(Poaceae), 8 October 2014, E. Camporesi, Holotype (MFLU
15–0656), Isotype (KUN), ex–type culture (MFLUCC 15–
0413, KIBCC, CMGCC).
Notes: Tristratiperidium microsporum is characterized by
semi-immersed, coriaceous ascomata, unitunicate, ellipsoidal-
subglobose, brown, uniseriately arranged, aseptate ascospores
with longitudinal germ slit and cylindrical asci with a J- apical
apparatus. Tristratiperidi um microsporum clustered with
Subramaniomyces fusisaprophyticus, which is morphologi-
cally distinct (Fig. 1). Subramaniomyces fusisaprophyticus is
a hyphomycete, described by Mani Varghese and Rao (1980)
and its sexual morph is unknown. However, the asexual char-
acters indicate that thesetaxaarenotcongeneric.
Subramaniomyces fusisaprophyticus has polyblastic conidia
produced in branched acropetal chains on mononematous,
branched conidiophores (Bhat and Sutton 1985). Unlike
S. fusisaprophyticus, Tristratiperidium microsporum has hya-
line, solitary conidia, produced terminally on conidiophores,
with phialidic conidiogenous cells. The septate conidia in
Tristratiperidium microsporum are hyaline and bear polar
setulae, which is quite unlike those of Subramaniomyces
fusisaprophyticus (Bhat and Sutton 1985).
Fig. 2 Tristratiperidium microsporum (holotype) a, b appearance of
ascoma on dead leaves of Arundo plinii, c perithecium in cross section,
d peridium in cross section, e clypeus, f–h asci, i ascospores in asci
showing straight germ slit, j, k immature ascospores, l, m mature
ascospores, n, o ascus apex in Lugol solution showing inamyloid (J-)
apical apparatus, p germinating ascospore. Scale bars; a = 500 μm,
b = 200 μm, c = 100 μm, d = 20 μm, e = 50 μm, f–i=20 μm, j–
m=10μm, n–o=10μm, p = 15 μm
Fig. 3 T ristratiperidium microsporum in OA after 2 weeks (from ex–
type culture MFLUCC 15–041 3) a from above, b from below, c
development of conidiomata in the culture, d, e conidiophores bearing
conidia, f–h conidia bearing setulae. Scale bars; d–h=10μm
8 Page 6 of 8 Mycol Progress (2016) 15:8
Discussion
The phylogenetic analyses (Fig. 1) revealed 11 distinct fami-
lies in Xylariales. The topology of well-supported branches is
similar in both single genes analyses (not shown) as well as in
combined gene analysis. This study, as well as in the previous
studies (Maharachchikumbura et al. 2015; Senanayake et al.
2015; Smith et al. 2003) was unable to provide strong support
for familial relationships of several clades within the order,
thus several families themselves are not clearly supported.
Many of the data available publicly only consist of few gene
sequences for a limited number of authentic strains of
Xylariales. Even a much smaller number of sequence data is
available for protein coding genes such as RPB2 or β-tubulin,
thus the interpretation of phylo genetic relationships in
Xylariales is not yet sufficiently informative and several line-
ages remain unresolved (Jaklitsch and Voglmayr 2012).
The sexual state of Tristratiperidium micr osporum shares
characteristics with certain taxa of Xylariales, especially those
with small, dark-colored, ascospores bearing longitudinal germ
slits. The asci and ascospores are similar to those of several
genera of microfungi of Xylariaceae like Anthostomella Sacc.,
Brunneiperidium Daranagama et al., Emarcea Duong et al.,
Helicogermslita Lodha & D. Hawksw., and
Str omatone ur o spora S.C. Jong & E.E. Davis. However, they
differ in stromatal morphology as Anthostomella and
Brunneiperidium have shiny black, carbonaceous, conical,
ostiolate ascomata with a peridium comprising two layers as
well as the latter two has cylindrical asci with long stipes with
J+ apical apparatus and the ellipsoidal-subglobose ascospores
in Tristratiperidium are smaller than those found in
Anthostomella and Brunneiperidium (Da ranagama et al.
2015). Emarcea also have immersed, solitary, papillate and
ostiolate ascomata reminiscent to T ristratiperidium but differs
from having multi-layered (4–6) peridium, long stipitate asci
with J+ apical apparatus bearing hyaline, fusiform, bicellular
ascospores (Duong et al. 2004). T ristratiperidium differs from
Helicogermslita by having dark brown-black, broadly ellipsoi-
dal ascospores with germ pores and helical germ slit with 2–4
coils (Hawksworth and Lodha 1983). Stromatoneurospora has
fusiform ascospores with longitudinally arranged ridges and
without germ slits unlike those of Tristratiperidium (Jong and
Davis 1973).
While the sexual characters of Tristratiperidium
microsporum are similar to those of other taxa of microfungi,
the asexual ch aracters differ considerably. The c onidia of
T. microsporum have polar, recurved setulae while in
Anthostomella species the asexual morph comprises hyaline,
aseptate, ellipsoidal conidia in branched hyaline conidio-
phores (Daranagama et al. 2015). The asexual state of
Tristratiperidium microsporum is similar to those of
Dictyochaeta Spegazzini,MenisporaPersoon, and Thozetella
Kuntze, but differs in having mononematous, dark brown,
unbranched, straight conidiophores, terminating with
percurrently proliferating, phialidic conidiogenous cells and
in the length and angle of insertion of setulae in the conidia.
Dictyochaeta has mononematous, branched or unbranched
conidiophores, mono- to polyphialidic conidiogenous cells
and aseptate conidia with polar setulae (Bhat and Kendrick
1993). Menispora has branched, septate and brown conidio-
phores bearing subhyaline phialides, with curved apices and
multi-sep tate conidia with or without long setulae (Seifert
et al. 2011). Thozetella has sporodochial conidiomata,
branched conidiophores with hyaline phialides and falcate,
hyaline and setulate conidia. Thozetella is further differentiat-
ed by presence of microawns in the sporodochial conidiomata
(Seifert et al. 2011).
Conidia with angled, short, and polar setulae have so far
not been observed in any other asexual state in Xylariales. In
addition, the presence of mononematous, dark brown, un-
branched, straight conidiophores terminating with
percurrently proliferating, phialidic conidiogenous cells pro-
ducing granulate, septate conidia with polar, recurved, setulae,
provides unique characteristi cs to differentiate the asexual
state of Tristratiperidium micr osporum from other hyphomy-
cetous asexual genera of Xylariales such as Dematophora R.
Hartig., Dicyma Boulanger, Geniculosporium Chesters &
Greenh., Nodulisporium Preuss, Virgaria Nees, Xylocladium
P. Syd. ex Lindau and Xylocoremium J.D. Rogers. All these
mentioned asexual genera except Xylocladium have branched,
brown-hyaline conidiophores with sympodial conidiogenesis
cells with apically aggregated scars bearing brown-hyaline
simple conidia without appendages while Xylocladium has
aspergilloid, brown conidiophores with a clavate vesicle and
sympodial, pale brown, vesiculate conidiogenesis cells with
hyaline, single conidia (Seifert et al. 2011).
Acknowledgments The authors appreciate the financial support and
postgraduate scholarship provided by State Key Laboratory of Mycology,
Institute of Microbiology, Chinese Academy of Sciences, Beijing and the
Mushroom Research Foundation, Chiang Mai, Thailand. The authors
gratefully thank Dr. Shaun Pennycook from Landcare Research Univer-
sity of Auckland, New Zealand for nomenclature advice on the proposed
names. The authors extend their sincere appreciations to the Deanship of
Scientific Research at King Saud University for its funding this Prolific
Research group (PRG-1436-09).
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