Phaeostilbelloides and Velloziomyces — new dematiaceous genera from the Brazilian Cerrado

Abstract

Phaeostilbelloides velloziae gen. & sp. nov. and Velloziomyces ramosiconidialis gen. & sp. nov. are described and illustrated. Both new fungi infect Vellozia squamata, endemic to the Brazilian Cerrado.

Full text

ISSN (print) 0093-4666 © 2015. Mycotaxon, Ltd. ISSN (online) 2154-8889
MYCOTAXON
http://dx.doi.org/./.
Volume 130, pp. 757–767 July–September 2015
Phaeostilbelloides and Velloziomyces —
new dematiaceous genera from the Brazilian Cerrado
E A  S A,
Z M C,  J C D*
Departamento de Fitopatologia, Universidade de Brasília,
Campus Darcy Ribeiro, Asa Norte, 70910-900 Brasília, DF, Brazil
* C : jcarmine@gmail.com
A —Phaeostilbelloides velloziae gen. & sp. nov. and Velloziomyces ramosiconidialis
gen. & sp. nov. are described and illustrated. Both new fungi infect Vellozia squamata,
endemic to the Brazilian Cerrado.
K  — neotropical fungi, asexual ascomycete morphs, mycodiversity, Velloziaceae
Introduction
Starting in 1993, an extensive inventory of fungi associated with plants
native to the Brazilian savanna, designated as the Cerrado, (Dianese 2000,
Dianese et al. 1993b, 1997) yielded important records of unusual hyphomycetes
such as those inhabiting the trichomes of Cerrado plants (Cantrell et al. 2011,
Pereira-Carvalho et al. 2009b). To date, 13 new ascomycete genera have been
described, ten as asexual morphs (Dornelo-Silva & Dianese 2004, Pereira-
Carvalho et al. 2009b) and three as sexual morphs (Dianese & al. 2001, Pereira-
Carvalho 2009a, Pereira-Carvalho et al. 2010). In addition, two new genera
of rust fungi, Batistopsora and Kimuromyces (Cummins & Hiratsuka 2003,
Dianese et al. 1993a, 1995), have been described and a third, Mimema, has
been reinstated (Cummins & Hiratsuka 2003, Dianese et al. 1994). Several
cercosporoid fungi (Dianese et al. 2008, Dornelo-Silva et al. 2007, Furlanetto
& Dianese 1999, Hernandez-Gutierrez & Dianese 2008, 2009, 2014, Inacio
& Dianese 2006, Medeiros & Dianese 1994) have been described and other
new ascomycetes have been included in other publications (Dornelo-Silva &
Dianese 2003, Inacio et al. 2011, 2012, Soares & Dianese 2014, Souza et al.
2008). is paper expands the number of genera among the asexual ascomycete

758 ... Armando, Chaves, & Dianese
morphs from the Cerrado, with the description and illustration of two new
genera, Phaeostilbelloides and Velloziomyces.
Materials & methods
Fungi associated with plants native in the Cerrado Biome were extensively collected.
Observations under a stereomicroscope revealed the presence of dematiaceous
hyphomycetes on leaves of Vellozia squamata Pohl (Velloziaceae), which is a Brazilian
Cerrado endemic. Squash preparations were mounted on semi-permanent slides stained
with lacto-glycerol/cotton blue or glycerol-KOH/basic phloxine. Mounting media
without stains were also used to determine the color of dierent structures. Sections
about 15–25 µm thick were produced using a Leica CM 1520 freezing microtome.
e structures observed in the squash preparations and sections were measured,
described, and documented using a Leica DM 2500 microscope coupled with a Leica
DFC 490 digital camera connected to a microcomputer. Image capture, editing, and
measurements were made with Leica QWin V3 soware. Whenever possible 20–50
measurements were made and expressed as range of length by width, with the mean
measures indicated in parentheses.
Taxonomy
Phaeostilbelloides Armando, Z.M. Chaves & Dianese, gen. nov.
MB MB
Diers from Phaeostilbella by its lack of rigid setae on the conidiomata.
T : Phaeostilbelloides velloziae Armando et al.
E: Latin Phaeostilbelloides, meaning similar to the genus Phaeostilbella.
C synnematous, scattered, erect, determinate, dark brown to
brown. C macronematous, cylindrical, branched just below
the apex, septate, straight or exuous, brown. C 
monophialidic, integrated, terminal, determinate. C acrogenous,
cylindrical to ellipsoid, unicellular, brown or pale brown.
Phaeostilbelloides velloziae Armando, Z.M. Chaves & Dianese, sp. nov. P 
MB MB
Diers from Phaeostilbella species by its lack of rigid curved setae on the conidiomatal
top and diers from Myrothecium species by its dark brown or brown conidiophores and
conidiogenous cells and its lack of a sporodochial phase.
T: Brazil. Distrito Federal: Brasília, West Sector of the IBGE-Roncador Reserve, on
living leaves of Vellozia squamata, 5 Apr. 2001, J.C. de Castro 82 (Holotype, UBMC
18404).
E: Latin velloziae, referring to the host genus.
L necrotic, linear, elongate, 5-10 × 0.5 mm, following parallel leaf
veins. M immersed;  light brown, septate - µm wide.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 759
F 1 Phaeostilbelloides velloziae (holotype, UBMC 18404) on leaf of Vellozia squamata.
A. Synnema showing stipe with textura intricata; B. Synnema top showing subapical branching
(white arrows), and abundant hyaline proliferations of the conidiophores (black arrow);
C. Conidiogenous cells and conidia; D. Conidia; E. Conidia with clearer area (arrow) at the conidial
base. Scale bars: A = 50 µm; C = 10 µm; B, D, E = 5 µm.
C 194-(212)-294 × 54-(56)-60 µm, synnematous cylindrical,
stipe textura intricata, smooth, dark brown, expanded to a 150-(166)-175 µm
wide brush-like capitulum, containing large number of immature
conidiophores that show up in squash preparations. C 3-5 µm

760 ... Armando, Chaves, & Dianese
in diam., cylindrical, smooth, originating as sub-terminal branches at
the capitulum, reduced to a single cell bearing two conidiogenous cells.
C  monophialidic, integrated, terminal, cylindrical,
10-(15)-20 µm long, 1-3 µm in diam., showing clear periclinal thickenings but
not a collarette. C 8-(7.5)-11 × 2-(2.5)-3 µm, noncatenate, aseptate,
sub-cylindrical, with an obtuse apex and truncate base, brown to dark brown,
with a slightly clear band just above the hilum, accumulating in mucilaginous,
dark brown masses.
A  : BRAZIL. D F: ,
Reserva Ecológica Águas Emendadas, on living leaves of Vellozia squamata, 25 Mar.
1995, Mariza Sanchez 530 (UBMC 7668).
C – Following the concepts by Seifert (1985) and Seifert et al. (2011),
it is not possible to place the specimen in any of the known genera producing
synnematous conidiomata, 1-celled brown to dark-brown conidia, and
phialidic conidial ontogeny. e closest genus, Myrothecium Tode [= Solheimia
E.F. Morris] as described and illustrated by Seifert et al. (2011), shares some
characteristics in common with the new taxon. However, the dierences between
Phaeostilbelloides velloziae and Myrothecium species are sucient to place
them in two separate genera. All Myrothecium species produce dark smooth
conidia, except for one species that shows conidia with strongly striate surface
and fusiform with acute ends. In addition, the synnematous conidiomata are
unbranched, and sporodochial conidiomata may also be present. In contrast,
P. velloziae lacks sporodochia, produces brown to dark brown smooth-walled
cylindrical conidia with an obtuse apex and a pale area just above the truncate
base, and synnemata that are sometimes branched. Another synnematous
species described from Brazil, Chaetantromycopsis bambusae H.P. Upadhyay et
al., produces black slimy spore heads but clearly diers from Phaeostilbelloides
by its hyaline to creamy white conidiophores and the presence of erect acute
hyaline to creamy white setae encircling the base of the synnemata (Upadhyay
et al. 1986). Finally, the Phaeostilbelloides type species is easily separated from
Phaeostilbella species because the hyaline cylindrical conidiophore branches
seen at the top in squash preparations dier completely from the rigid slightly
curved appendices of Phaeostilbella. Furthermore, Phaeostilbella synnemata
typically show textura prismatica, not textura intricata as in Phaeostilbelloides.
Also, the only species currently accepted in Phaeostilbella was reported on
grasses in Europe (Seifert et al. 2011) while Phaeostilbelloides velloziae is present
in a dicotyledonous host endemic to the Cerrado.
e dichotomous key below, modied from Seifert et al. (2011), segregates
Phaeostilbelloides from all other genera with synnematous conidiomata forming
phialidic conidiogenous cells bearing 1-celled conidia.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 761
Key to the dematiaceous genera of phialidic synnematous anamorphs
forming 1-celled conidia
1. Conidia hyaline, globose to ellipsoidal, ovoid, fusoid,
in clear or bright colored mucilaginous false heads..........................2
1. Conidia brown to dark brown, cylindrical, fusoid,
in dark brown to black false heads........................................7
2. Synnemata originating from broad well developed stromata, found on wood
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Stromatographium Höhn.
2. Synnemata lacking a well-developed basal stroma..............................3
3. Synnemata with ornamenting cells on stipe
. . . . . . . . . . . . . . . . . . . . . anamorphic Stilbocrea Pat. (= Gracilistilbella Seifert)
3. Synnemata without ornamenting cells on the stipe .............................4
4. Synnematous heads with sterile hyphae among the conidiogenous cells
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .anamorphic Holwaya Sacc. (= Crinula Fr.)
4. Synnematous heads lacking sterile hyphae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5
5. Conidiophores penicillate occurring in pustules on the upper portion of the stipe
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cornutostilbe Seifert
5. Stipe without conidiophores in pustules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6
6. Conidiogenous cells lageniform with characteristically narrow elongated necks,
conidia obovoid.....................................Dennisographium Rifai
6. Conidiogenous cells holoblastic or phialidic terminal at the tip of
each conidiophore, lacking narrow neck...........Synnemellisia N.K. Rao et al.
7. Synnemata with setose capitulum, showing a crown of dark setae,
conidia fusoid ........................................Phaeostilbella Höhn.
7. Synnemata lacking setae at capitulum or with subhyaline hyphae . . . . . . . . . . . . . . . .8
8. Capitulum without sterile hyphae, conidia fusiform, with smooth or striate surface,
sporodochial phase oen present..................Myrothecium (= Solheimia)
8. Capitulum non-setose but with subhyaline hyphae, conidia cylindrical,
with obtuse apex and pale band just above the truncate base,
sporodochial phase absent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Phaeostilbelloides
Velloziomyces Armando, Z.M. Chaves & Dianese, gen. nov.
MB MB
Diers from Minimidochium by its holoblastic conidial ontogeny and from
Dictyodochium by its schizolytic conidial secession.
T : Velloziomyces ramosiconidialis Armando et al.
E: From the Latin Velloziomyces for its occurrence on Vellozia.
C sporodochial scattered or conuent, setose, dark brown, S
cylindrical, erect, straight or exuous, continuous or septate, brown to dark
brown. C undierentiated C  monoblastic,
cylindrical to lageniform, indeterminate, with several enteroblastic percurrent

762 ... Armando, Chaves, & Dianese
extensions, integrated, terminal. C solitary, obclavate or Y-shaped,
0-2-septate, simple or branched, brown to pale brown.
Velloziomyces ramosiconidialis Armando, Z.M. Chaves & Dianese, sp. nov.
MB MB P 2, 3
Diers from Minimidochium species by its holoblastic conidial ontogeny, from
Dictyodochium species by its schizolytic conidial secession of phragmospores, and from
Iyengarina species by its setose sporodochial conidiophores and association with living
leaves.
T: Brazil. Distrito Federal: Planaltina, Estação Ecológica Águas Emendadas, on
living leaves of Vellozia squamata, 13 Jun. 1995, Arenildo Soares Alves 215 (Holotype,
UBMC 8849).
E: From the Latin ramosiconidialis, referring to conidial branching.
C amphigenous, light brown, erumpent, setose. Mycelium mostly
immersed, composed of septate, light brown, smooth hyphae. C
sporodochial suprastomatal, scattered or conuent, setose, dark brown,
10-(42)-55 µm diam. S cylindrical, erect, straight or exuous, sometimes
slightly curved, obtuse or acerose at the apex, continue or septate, brown to dark
brown, 60-(112)-138 × 3-(4)-6 µm. C  monoblastic,
indeterminate, with several enteroblastic percurrent extensions, terminal,
short cylindrical, lageniform 3-9 × 3-5 µm. C solitary, 1-2-septate,
more or less Y-shaped, with a lateral branch, obclavate at main axis (7-(18)-25
× 3-(8)-16 µm) and one shorter lateral branch, light brown, with paler conical
apical cell, truncate base.
A  : BRAZIL. D F: B, Jardim
Botânico de Brasília, on living leaves of Vellozia squamata, 24 May 2011, Eliane
Armando 129 (UBMC 21928).
C – Sporodochial conidiomata are distributed in several genera
of dematiaceous anamorphs (Ellis 1971, Ellis 1976, Kier and Morelet 2000,
Seifert 1985, Seifert et al. 2011). However, there is no published genus described
as producing setose sporodochia on top of an obconical stromatic structure
that grows through the stomata. As illustrated here, the stroma originates
deep within the leaf parenchyma, grows through the stoma to expand at the
surface into a multicellular plate formed by a layer of tightly packed annellidic
conidiogenous cells surrounded by dark brown elongated setae. In addition, the
light brown conidia are unusual because at maturity they have one unicellular
conical branch growing from the basal cell, which is shorter than the main
2–3-celled axis of the spore. Sometimes the branch is not present.
Two genera, Minimidochium B. Sutton and Dictyodochium Sivan., as shown
in Seifert et al. (2011), have setose sporodochial conidiomata that originate
from internal mycelium emerging from plant tissue, as in Velloziomyces.

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 763
F  Velloziomyces ramosiconidialis (holotype, UBMC 8849) on leaf of Vellozia squamata.
A. Setose stromatic sporodochium; B. Sporodochium detail of the supercial distribution of the
conidiogenous cells; C. Setose sporodochium bearing conidia; D. Setae detail and a group of free
conidia; E. Setose sporodochium; F. Conidiogenous cell; G–H. Conidia focused in dierent depths.
Scale bars: A = 20 µm; B–E = 10 µm; F–H = 5 µm.
However, all eight species of Minimidochium: M. crepuscolare C. Ciccar.,
M. eucalypti Vittal & Dorai, M. indicum Varghese & V.G. Rao, M. microsporum
Matsush., M. monoseptatum G.C. Zhao, M. nipponicum Matsush., M. parvum

764 ... Armando, Chaves, & Dianese

Phaeostilbelloides & Velloziomyces gener. nov. (Brazil) ... 765
F 3 Velloziomyces ramosiconidialis (holotype, UBMC 8849) on leaf of Vellozia squamata
(SEM). A–C. Developing sporodochium seen from the top; D. Sporodochium showing a mature
seta; E. Setose sporodochium and conidiogenous cells (arrows), and a conidium (C) still attached
to the conidiogenous cell; F. Top view of a typical setose sporodochium. Scale bars: A–E = 10 µm;
F = 40 µm.
Cabello et al., and M. setosum B. Sutton (Cabello et al. 1998, Ciccarone 1988,
Matsushima 1995, Sutton 1969, Varghese & Rao 1980, Vittal & Dorai 1991, Zhao
& Zhao 2012) produce phialidic conidiogenous cells bearing typically setulose
hyaline conidia. On the other hand, in the monotypic genus Dictyodochium,
the muriform brown conidia have rhexolytic secession. Also, the three species
belonging in Iyengarina Subram. (Kuthubutheen & Nawawi 1992, Subramanian
1958) are easily segregated from V. ramosiconidialis due to clear dierences in
conidial morphology, inability to form setose sporodochial conidiomata, and
association with plant debris instead of living leaves. us, species belonging in
all three genera clearly dier from V. ramosiconidialis, which is characterized by
its holoblastic annellidic conidiogenous cells, non-setulose branched conidia
showing schizolytic secession, and setose sporodochial conidiomata.
Acknowledgements
e authors thank Drs. R.F. Castañeda-Ruiz (INIFAT, Cuba) and Paul Kirk
(Kew Gardens, UK) for critical commentaries that greatly improved our manuscript
and CNPq-Brazil for the graduate fellowship provided to the senior author and for
nancing the execution of the entire research project through funds provided by the
PPBIO-Cerrado/CNPq. Finally, gratitude is expressed to Prof. Mariza Sanchez for
assistance and friendship.
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