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3852/12-137
Short title: Phialemoniopsis gen. nov. and new species of Phialemonium and Lecythophora
Lecythophora
Haybrig Perdomo
Dania García
Josepa Gené1
Josep Cano
Mycology Unit, Medical School and IISPV, Universitat Rovira i Virgili, Sant Llorenç 21,
43201-Reus, Spain
Deanna A. Sutton
Richard Summerbell
Sporometrics, Inc. 219 Dufferin Street, Suite 20C, Toronto, Ontario, M6K 1Y9, and Dalla
Lana School of Public Health, University of Toronto, 223 College Street, Toronto Ontario,
M5T 1R4
Josep Guarro
Mycology Unit, Medical School and IISPV, Universitat Rovira i Virgili, Sant Llorenç 21,
43201-Reus, Spain
clarify the phylogenetic relationships among these fungi and other related taxa, we performed
a polyphasic study based on a detailed morphological study and on the analysis of sequences
of four loci: the internal transcribed spacer regions, the D1/D2 domains of the 28S rRNA,
actin and β-tubulin genes. The combination of the resulting data let us propose the new genus
Sarcopodium oculorum and two new species, Phialemoniopsis cornearis and Phialemoniopsis
pluriloculosa. The taxonomy of Phialemoniopsis has not been completely resolved, however,
remaining incertae sedis within the Sordariomycetes. In addition, the new species
described and the species Acremonium atrogriseum and Taifanglania inflata are transferred to
the genus Phialemonium. Lecythophora and Phialemonium are currently monophyletic genera
provided.
INTRODUCTION
Phialemonium W. Gams & McGinnis is an anamorphic genus with species widely distributed
and commonly isolated from air, soil, industrial water and sewage (Gams and McGinnis
1983). They also are involved in human infections, affecting both immunocompromised and
immunocompetent patients (King et al. 1993, Guarro et al. 1999, de Hoog et al. 2000, Proia et
al. 2004, Scott et al. 2004, Rivero et al. 2009). Phialemonium, typified by P. obovatum W.
Gams & McGinnis, was described by Gams and McGinnis (1983) to accommodate
filamentous fungi with morphological features intermediate between Acremonium Link and
Phialophora Medlar. The species of Phialemonium are characterized by moist, flat, never
pink colonies and abundant adelophialides (reduced phialides that are not delimited from
subtending intercalary hyphal cell by a basal septum), although short discrete phialides
without a collarette also are present. The phialides form unicellular, hyaline conidia
aggregated in slimy heads (Gams and McGinnis 1983). Currently, the genus Phialemonium
comprises two species, P. obovatum and P. curvatum W. Gams & W.B. Cooke.
sympodially proliferating and obovate conidia with an apiculate and minutely truncate base
grayish colonies, non-proliferating phialides and cylindrical to allantoid conidia (Gams and
McGinnis 1983, King et al. 1993, de Hoog et al. 2000). Some authors have described the
original description (Proia et al. 2004, Strahilevitz et al. 2005, Weinberger et al. 2006, Sutton
Yaguchi et al. (2006) described the new species Cephalotheca foveolata Yaguchi,
Nishim. & Udagawa from a patient with a subcutaneous infection, which showed a
Phialemonium-like anamorph. In the same study, analysis of the LSU rDNA sequences
curvatum into Cephalothecaceae was not supported by the sequence analysis. Similar results
were found by Perdomo et al. (2011b), who demonstrated that Phialemonium is not
Syd. and Neolinocarpon K.D. Hyde, two ascomycetous genera that are incertae sedis within
characterized by the production of pigmented conidia arranged in chains and slimy heads, is
This is a rare opportunistic species described as agent of a human systemic infection and
repeatedly isolated from different clinical sources (de Hoog et al. 2000, Perdomo et al.
2011a). Other fungi show some morphological resemblance and are genetically related to
the new genus Taifanglania Z.Q. Liang, Y.F. Han, H.L. Chu & R.T.V. Fox (Liang et al. 2009)
and some species of Sarcopodium Ehrenb. and Volutella Tode, such as S. oculorum Gené &
Guarro and V. cinerescens (Ces.) Sacc. The latter two species were described as agents of
ocular infections (de Hoog et al. 2000, Guarro et al. 2002, Weinberger et al. 2006).
adelophialides with collarettes, although discrete phialides like those of Acremonium also may
be present (de Hoog et al. 2000, Weber 2002). Lecythophora Nannf. is a cosmopolitan genus,
which comprises six species: L. lignicola Nannf. (type species), L decumbens (J.F.H. Beyma)
E. Weber, Görke & Begerow, L. fasciculata (J.F.H. Beyma) E. Weber, Görke & Begerow, L.
hoffmannii (J.F.H. Beyma) W. Gams & McGinnis, L. luteoviridis (J.F.H. Beyma) E. Weber,
Görke & Begerow, and L. mutabilis (J.F.H. Beyma) W. Gams & McGinnis. The
morphological characters used to differentiate these species are the presence in L. mutabilis
and L. luteoviridis of chlamydospores, which are ovoidal to ellipsoidal and solitary in the
former and globose to oblong or pear-shaped and solitary or in short chains in the latter; the
developing ventricose phialides and adelophialides. Lecythophora spp. have been isolated
from wood, water and food (Weber 2002), and some species are also known as human
pathogens. Lecythophora hoffmannii and L. mutabilis have been isolated from keratitis,
subcutanous abscesses, peritonitis, endocarditis and septic shock (de Hoog et al. 2000,
Lecythophora are Cadophora Lagerb. & Melin, Collophora Damm & Crous, Hormonema
Lagerb. & Melin, Hyphozyma de Hoog & M.T. Sm., Margarinomyces Laxa,
Phaeoacremonium W. Gams, Crous & M.J. Wingf. and Phialophora. These genera belong to
ascomycete orders other than those embracing Lecythophora and Phialemonium (Seifert et al.
2011).
Lecythophora species, analyzing the D1/D2 domains of the 28S rRNA gene sequences, we
found that some isolates could not be properly identified to species and were considered to be
possible cryptic species (Perdomo et al. 2011b). In the present study, to clarify the
phylogenetic relationships among these fungi and other related or morphologically similar
taxa, as well as to describe new species, we performed a multilocus sequence analysis of the
internal transcribed spacer (ITS) regions, the D1/D2 domains of the 28S rRNA (D1/D2), actin
characterization of the isolates, re-evaluating the characters seen in the light of the
Isolates and morphology.—Twenty-four clinical isolates from the Fungus Testing Laboratory, Department of
Pathology, University of Texas Health Science Center at San Antonio (UTHSC) were included in the present
study. Twenty-two isolates were identified in Perdomo et al. (2011a, b) as Acremonium atrogriseum (n = 1),
Phialemonium sp. 1 (n = 5), Phialemonium sp. 2 (n = 1) and Lecythophora sp. 1 (n = 2). Two other clinical
isolates were received as Volutella sp. (UTHSC 04-7 and UTHSC 09-3589). In addition, type or reference strains
of Phialemonium and Lecythophora spp., and species of other morphologically similar genera, such as
Sarcopodium oculorum, S. circinatum Ehrenb. , Volutella ciliata (Alb. & Schwein.) Fr., V. cinerescens,
Cephalotheca sulfurea Fuckel and Taifanglania inflata (Burnside) Z.Q. Liang, Y.F. Han & H.L. Chu, also were
For morphological characterization, the isolates were examined on oatmeal agar (OA: 30 g oat flakes,
20 g agar, 1L distilled water), potato dextrose agar (PDA, Difco Laboratories, Detroit, Michigan) and potato
carrot agar (PCA: 20 g potatoes; 20 g carrot; 20 g agar; 1 L tap water). Water agar with sterilized plant material
(small pieces of wood, filter paper and leaves of herbaceous plants) also was used to enhance the formation of
ascomata or conidiomata. Cultures were incubated at room temperature (25 C ± 2 C) in the dark, up to 6 mo.
Growth rates of the isolates were determined on PDA plates at 15, 25, 30, 35, 37, 40 and 42 C for 14 d in
darkness. In descriptions color colony codes in parenthesis refer to Kornerup and Wanscher (1978). The
microscopic features were examined and mesured in lactic acid under a light microscope (Olympus CH-2).
Photomicrographs were obtained with a Zeiss AXIO Imager M1 light microscope and a Jeol JSM-6400 scanning
electron microscope.
Molecular study.—DNA was extracted according to Perdomo et al. (2011a). Protocols for the amplification of
the ITS, D1/D2, ACT and TUB sequences follow Cano et al. (2002), Voigt and Wöstemeyer (2000), and
Gilgado et al. (2005). PCR products were purified according to the manufacturer's instructions using a
commercial kit (Illustra GFX™ PCR DNA and Gel Band Purification, General Electric Healthcare,
Buckinghamshire, UK). Sequencing reactions were carried out with the PCR primers using a Taq DyeDeoxy
Terminator Cycle Sequencing Kit (Applied Biosystems, Gouda, the Netherlands), according to the manufacter's
recommendations, and the resulting products were analyzed on a 310 DNA sequencer (Applied Biosystems).
DNA sequences were aligned with the program Clustal X 1.8 (Thompson et al. 1997). In addition to the
sequences of the clinical isolates and those of type and reference strains generated in this study, several
sequences of other related taxa were retrieved from the GenBank for comparison in this analysis.
The relationships of the fungi tested in this study at suprageneric rank were determined first through the
analysis of D1/D2 domains sequences. Then we performed a multilocus sequence analysis of the D1/D2
domains, ITS regions and a fragment of the ACT and TUB genes of those clades obtained in the first analysis
Maximum likelihood (ML) analysis of the D1–D2 sequences was conducted with MEGA 5.05 (Tamura
et al., 2011), using Tamura Nei substitution model with gamma distribution. Support for internal branches was
assessed by a search of 1000 bootstrapped sets of data. Bayesian analysis was performed with MrBayes3.0
(Huelsenbeck and Ronquist 2001, Ronquist and Huelsenbeck 2003) by running 1 000 000 generations in four
chains, saving the current tree every 100 generations. The last 10 000 trees were used to construct a 50%
majority rule consensus tree and to determine the posterior probabilities of the branches. The tree was conducted
In the multilocus study, most parsimonious tree (MPT) analyses of the combined dataset of the loci
mentioned above were performed with PAUP* 4.0b10 (Swofford 2002). One hundred heuristic searches were
conducted with random sequence addition and tree bisection reconnection branch-swapping algorithms,
collapsing zero-length branches and saving all minimal-length trees (MulTrees) on different datasets. The gaps
were treated as missing data and support for internal branches was assessed with a heuristic parsimony search of
1000 bootstrapped datasets. The combined dataset was tested for incongruence with the partition homogeneity
test (PHT) as implemented in PAUP*. Tree length, consistency index (CI), retention index (RI) and the
homoplasy index (HI) values also were calculated. In those analyses the sequences of Lecythophora lignicola
(HE610463 of CBS 267.33) or Phialemonium obovatum (FR691997 of CBS 279.76) were used as outgroup.
The sequences generated in this study and the alignments used the phylogenetic analyses were
RESULTS
Phylogenetic analyses.—We illustrated the phylogenetic tree inferred from the analysis of the
genetically related (FIG. 1). The aligned sequence regions consisted of 469 bp. The sequences
were distributed in three main clades, that is clade I, incertae sedis (99% bs and 1 pp), clade
II, Hypocreales (91% bs and 1 pp) and clade III, Sordariales (clade III, 76% bs and 1 pp),
while that the representatives of Coniochaetales grouped with low bootstrap support (bs).
Clade I embraced 12 clinical isolates, the type and reference strains of P. curvatum, one
reference strain of Volutella cinerescens (CBS 832.71), the type and a reference strain of
Sarcopodium oculorum (IHEM 19077, FMR 7190 respectively) and GenBank sequences of
K.D. Hyde and Neolinocarpon enshiense K.D. Hyde, Joanne E. Taylor & J. Fröhl. Because
the type species of the genus Phialemonium (P. obovatum) was distant from clade I and
included in the order Sordariales (clade III), clade I may represent an anamorph genus
different from Phialemonium. This clade is also different from Sarcopodium or Volutella
because reference strains of the type species of these two genera (S. circinatum, V. ciliata
respectively) were placed in the order Hypocreales (Clade II). Clade III included members of
the family Cephalothecaceae related to P. obovatum; that is type strains and clinical isolates
Malloch & Cain, Cryptendoxyla hypophloia Malloch & Cain and P. inflatus, reference strains
of T. inflata and C. sulfurea, in addition to the clinical isolate UTHSC 03-3661, which was
closely related to the latter three fungi and identified in Perdomo et al. (2011b) as
Phialemonium sp. 2. Finally, the type and reference strains of all the currently accepted
species of Lecythophora, some members of the ascomycetous genus Coniochaeta and the
clinical isolates UTHSC 01-20-1 and UTHSC 01-1644 identified in Perdomo et al. (2011b) as
To confirm the phylogenetic relationships in the D1/D2 domains tree and determine
the boundaries of some of the taxa included in the study, we performed separated multilocus
analyses of clades I, III and the Coniochaetales group, including the most representative
We illustrated the multilocus analysis of clade I (FIG. 2). Two MPT were produced
from a heuristic search using the combined dataset of 2135 characters from the four loci
tested. The resulting topology was similar to the topologies obtained by analyzing the genes
individually (data not shown). Analyses of the combined partitions support the recognition of
four well supported terminal clades similar to those previously observed (FIG. 1), each
(FMR 7190) of S. oculorum, and three additional isolates (UTHSC 05-2527, UTHSC 07-
3736, UTHSC 09-2358). Sequences variation within these isolates was 0–1 bp/454 total bp
for D1/D2 (99.3–100%), 1–8/476 for ITS (99.2–99.7%), 0–12/713 for ACT (98.2–100%) and
0–2/375 for the TUB (99.8–100%). The second and the third clades were composed of two
clinical isolates each (100% and 96% bs respectively). The fourth clade (100% bs) included
the type and reference strains of P. curvatum, and the strain V. cinerescens CBS 832.71.
Sequence similarities within this group were 99.4–100% for D1/D2, 99.2–100% for ITS,
The combined multilocus analysis of the Coniocahetales group included eight strains
of Lecythophora spp. and P. obovatum as outgroup (FIG. 3). The maximum parsimony
analysis of the dataset (2173 bp) produced one MPT, with eight terminal branches. Six
corresponded to known species and the other two, UTHSC 01-20-1 and UTHSC 01-1644,
are illustrated (FIG. 4). The phylogenetic analysis of the dataset (2175 bp) yielded one MPT,
in which three well supported (100% bs) primary clades, were shown. The first clade was
composed of two sister terminal subclades; one (100% bs) included the type and different
isolates of P. obovatum, and the other (100% bs) grouped the type strain and four clinical
isolates of C. foveolata. The second clade contained the type strain of A. atrogriseum and
three conspecific isolates (Perdomo et al. 2011a). The similarities among the sequences of the
members of this group were 99.8–100% for D1/D2, 98.6–100% for ITS, 98.5–99.9% for ACT
and 98.8–100% for the TUB. The third clade was divided into three terminal subclades; the
first one included a reference strain of T. inflata and the type strain of P. inflatus. The
sequence similarity was 98.9–100% for all the loci studied, which demonstrated that both
species are conspecific. The second subclade included only the clinical isolate UTHSC 03-
3661, and finally the third subclade was constituted by CBS 135.34, a reference strain of C.
Morphological study.—The morphological study of the 24 clinical isolates and numerous type
and reference strains of Phialemonium, Lecythophora and related taxa, let us characterize
phenotypically the different clades shown in the trees inferred from the phylogenetic analyses.
flexuose, hyaline to brown conidiogenous cells (discrete phialides and adelophialides), in all
cases with collarettes. They are solitary, borne directly on undifferentiated hyphae or
the conidiomatal types also were present in some cases. Four morphotypes could be
distinguished within theses isolates, which corresponded to the four main phylogenetic clades
observed in the multilocus analysis (FIG. 2). One morphotype included the three clinical
isolates identified as Phialemonium sp. 1 (Perdomo et al. 2011b) and the type and a reference
strain of S. oculorum; they all develop sporodochium- and pycnidium-like conidiomata. The
conidiomatal structures were characterized by a wall with two types of tissue, textura intricata
A second morphotype contained two isolates, UTHSC 06-1820 and UTHSC 06-1465.
A third morphotype included the isolates UTHSC 04-7 and UTHSC 09-3589. These
isolates have multilocular conidiomata, with a wall of textura epidermoidea, and ellipsoidal
conidia.
The last morphotype included the isolates identified as P. curvatum; three of them,
UTHSC 06-4324, UTHSC R-3447 and UTHSC R-3448, were characterized by the presence
of sporodochium-like conidiomata. These structures also were observed in the conspecific
strain CBS 832.71, which was received from the CBS collection as V. cinerescens.
All the members of clade III (FIG. 1) developed discrete phialides or adelophialides
with a cylindrical or slightly swollen basal region, usually without visible collarettes. These
characters are typical of the genus Phialemonium. Within this clade, we recognized six
morphotypes, which corresponded with the terminal clades shown in the corresponding
conidia with a truncate base and oval chlamydospores. Those in the second morphotype,
occasionally with a markedly inflated base, and produced slightly pigmented obovoid conidia
A fourth morphotype included the strains of T. inflata and P. inflatus, which showed
discrete phialides with a swollen basal part, tapering into a long neck, and citriform conidia.
showed phialides similar to those described in strains of A. atrogriseum and P. inflatus, but
globose conidia.
The sixth morphotype, which corresponded to the reference strain of C. sulfurea (CBS
135.34), developed an anamorph with flask-shaped phialides and ovate conidia. The conidia
in strains of the last-mentioned three morphotypes formed long and persistent chains, and they
had connectives on both ends. This character was not described previously for the genus
Most isolates included in the Coniochareales group (FIG. 1) showed the typical
UTHSC 01-1644, had characteristics that were distinct from those of the other described
species of Lecythophora. The first isolate, UTHSC 01-20-1, excreted a diffusible orange
pigment and produced discrete phialides and adelophialides with a swollen base, while the
TAXONOMY
The morphological features shown by the Phialemonium species in clade I are very different
from the typical features shown by Phialemonium s. str. and its relatives (clade III). Because
these morphological differences also are reflected in the multilocus sequence analysis, we
therefore consider that the fungi included in clade I represent a new genus, which is proposed
below as Phialemoniopsis. Four species are described in the new genus. Furthermore, three
new species of Phialemonium and two other of Lecytophoraare are described in the present
study.
MycoBank MB563874
Colonies growing moderately with aerial mycelium sparse or lacking, brown or grayish.
conidiophores solitary, borne on undifferentiated hyphae from the aerial mycelium or grouped
several layers; wall of textura intricata or textura epidermoidea or both, with an irregular
dehiscence (lacking apical preformed opening or ostiole), often appearing cupulate when
mature. Setae usually present, forming a frill at the margin of open conidiomata or
and slightly apiculate at the base, hyaline to subhyaline, smooth-walled, aggregated in slimy
Phialemoniopsis cornearis H. Perdomo, D. García, Gené, Cano & Guarro, sp. nov. FIG. 5
MycoBank MB563875
Colonies on OA at 25 C attained 54–60 mm diam after 14 d, flat, velvety, with whitish soft
cottony aerial mycelium and olive-brown (4F3) submerged hyphae; reverse colorless to dark
brown (6F5) with age. Colonies on PDA at 25 C, 35–40 mm diam after 14 d, fasciculate,
radially folded, initially white to brownish gray (4E2), becoming olive-brown (4E4) to black;
reverse colorless first becoming olive-brown (4E4). The fungus grew slowly at 37 C (up to 9
mm diam after 14 d); no growth at 40 C; optimal temperature for growth was at 25–30 C.
Vegetative hyphae initially hyaline, smooth-walled, becoming brown and verruculose, 2–4
Conidiophores from aerial mycelium mostly short, erect, often simple or few branched, 10–30
× 1–2 μm. Phialides and adelophialides straight to slightly flexuose, cylindrical, 2–24 × 1–2
μm, with conspicuous collarette, initially hyaline to subhyaline becoming brown, smooth-
walled. Conidia one-celled, ellipsoidal to slight allantoid, 4–7 × 1–2 μm, slightly apiculate at
subglobose, 300–650 × 200–450 μm; wall 3–4 μm thick, with textura intricata, dark brown to
black. These conidiomata open irregularly and often appearing cupulate when mature and
surrounded by subhyaline to brown setae. Setae forming a frill at the margin of the
μm long, 1–2 μm wide, hyaline, subhyaline, or brown, smooth and slightly thick-walled.
group of 3–5 discrete phialides. Phialides cylindrical or subcylindrical, 9–25 × 1–1.5 μm, with
slightly apiculate at the base, 2–3 × 1–1.5 μm, one-guttulate, smooth-walled, hyaline, pale
lateral, solitary or in short chains, globose, oval to pyriform, 5–9 × 4–5 μm, initially hyaline
Etymology: From the Latin cornearis referred to the clinical specimen from which the
Specimens examined: USA. FLORIDA. From human corneal fluid, D.A. Sutton. (CBS H-20781,
HOLOTYPE; UTHSC 06-1820, CBS 131711, FMR 10701, ex-type cultures). USA. SOUTH CAROLINA. From
the pycnidium-like conidiomata of P. cornearis shows a wall of textura intricata and it usually
Phialemoniopsis curvata (W. Gams & W. B. Cooke) H. Perdomo, D. García, Gené, Cano &
MycoBank MB563876
≡Phialemonium curvatum W. Gams & W. B. Cooke, in Gams & McGinnis, Mycologia 75:980 (1983).
white becoming brown (6E5) to grayish brown (6E3). On PDA at 25 C, 46–48 mm diam after
14 d, cream-colored (4A3), with small areas light brown (6D4). The fungus grew slowly at 37
C (5–11 mm diam in 14 d); no growth at 40 C; optimal temperature for growth was at 25–30
C. Vegetative hyphae hyaline to light brown, 1–2 μm wide, smooth- to slightly rough-walled.
conidiomata in some isolates (CBS 832.71, UTHSC 06-4324, UTHSC R-3447, UTHSC R-
3448). Adelophialides abundant, 4–12 × 1–1.5 µm, with barely visible collarettes. Discrete
phialides less common, long, cylindrical, slightly tapering toward the apex, 8–20 × 2–3.5 µm,
with barely visible collarettes. Conidia cylindrical or allantoid, 4–6 × 1–2 µm, hyaline,
conidiophores, with a marginal frill of setae and with a slimy conidial mass covering the
entire upper surface of the conidioma. Setae unbranched or slightly branched toward the base,
straight or flexuose, septate, cylindrical, with slightly pointed or blunt tip, up to 60 μm long,
culture). USA. CALIFORNIA: From chaparral soil, W.B. Cooke (CBS 491.82). USA. MINNESOTA: From
human sinus, D.A. Sutton (UTHSC 04-956). USA. TEXAS: From canine pleural fluid, D.A. Sutton (UTHSC 06-
4324). USA. UTAH. From human blood, D.A. Sutton (UTHSC 08-2292). ISRAEL. From human eye, D.A.
Sutton (UTHSC R-3447); from human eye, D.A. Sutton (UTHSC R-3448). As Volutella cinerescens: from
Phialemoniopsis curvata is mainly distinguished from the other species of the genus
Phialemoniopsis ocularis (Gené & Guarro) H. Perdomo, D. García, Gené, Cano & Guarro,
MycoBank MB563877
≡ Sarcopodium oculorum Gené & Guarro, in Guarro et al., J. Clin. Microbiol. 40:3074 (2002).
periphery, with whitish, soft cottony aerial mycelium and brownish gray (4E2) submerged
(4A3) to brownish gray (4E2), radially folded and granulose due to the abundant production
of conidiomata. The fungus grew slowly at 37 C (9–14 mm diam after 14 d); no growth at 40
C; optimal temperature for growth was 25 C. Sporulation from conidiophores solitary on the
Phialides and adelophialides from aerial mycelium straight to slightly flexuose, cylindrical, 3–
19(–36) × 1–2 μm, with collarettes, initially hyaline to subhyaline becoming brown, smooth-
walled. Conidia ellipsoidal to slightly allantoid, 3–5 × 1–1.5 μm, slightly appiculate at the
forming a frill at the margin of the conidiomata and interspersed with the conidiophores.
and epidermoidea. These conidiomata open by an irregular rupture and often appear cup-
shaped when mature and surrounded by setae. Setae erect, usually unbranched, straight or
flexuose, septate, cylindrical, with slightly pointed or blunt tips, up to 65 μm long, 1.5–2.5
bearing a group of 2–4 phialides terminally. Phialides usually with a slightly swollen basal
part and slightly tapering toward the tip, 5–8 × 1–1.5 μm, hyaline. Conidia one-celled,
ellipsoidal, some lightly allantoid, 2–3 × 1–1.5 μm, with a slightly apiculate base, one-
globose to pyriform, 5–9 × 3–6 μm, initially hyaline becoming brown, thick- and rough-
Specimens examined: BRAZIL. Sao Paulo: From human keratitis, P. Godoy (IHEM 19077, ex-type
culture). BRAZIL. Sao Paulo: From human keratitis, P. Godoy (FMR 7190). USA PENNSYLVANIA: From
peritoneal dialysis catheter, D.A. Sutton (UTHSC 05-2527). USA. FLORIDA: Left hand, D.A. Sutton (UTHSC
07-3736). USA. MASSACHUSETTS: From aspirate cellulitis, D.A. Sutton (UTHSC 09-2358).
A description of this species was given by Guarro et al. (2002). However, pycnidium-
from the aerial mycelium and with the sporodochium- and pycnidium-like conidiomata. The
main distinctive feature of P. ocularis is that the pycnidium-like conidiomata have a wall
made of both textura intricata and textura epidermoidea, and the conidia from these structures
are usually ellipsoidal. In addition, it differs from the other species by its growth at 37 C.
Phialemoniopsis pluriloculosa H. Perdomo, D. García, Gené, Cano & Guarro, sp. nov. FIG. 7
MycoBank MB563878
Colonies on OA 25 C attained 54–55 mm diam after 14 d, velvety and yellowish white (4A2),
but soon become olive-brown (4E4) to black; reverse colorless to black with age. On PDA at
25 C the colonies reaching 40–43 mm diam after 14 d, mucous, radially folded, yellowish
white (4A2); reverse pale yellow (4A3). The fungus grew slowly at 35 C (up to 8 mm diam in
initially hyaline, smooth-walled, becoming brown and slightly verruculose, 1–2 μm wide.
Sporulation from conidiophores growing on the aerial mycelium, and from sporodochium-
straight to slightly flexuose, cylindrical, 6–20 × 1–2 μm, with short collarettes, initially
cylindrical, some slightly curved, 3–5 × 1–2.5 μm, with a slightly apiculate base, biguttulate,
450 × 150–350 μm; wall 10–15 μm thick, with textura epidermoidea, dark brown to black,
non-ostiolate, opening irregularly and often appearing cupulate and with marginal setae. Setae
unbranched or slightly branched toward the base, straight or flexuose, septate, cylindrical,
with slightly pointed or blunt tips, up to 60 μm long, 2.5–3 μm wide, subhyaline to brown,
branched, and often proliferate sympodially showing conidiogenous cells as lateral cylindrical
projections, hyaline. Phialides subcylindrical, slightly tapering toward the tip, 6–12 × 1–1.5
μm, hyaline. Conidia one-celled, ellipsoidal, 2–3 × 1–1.5 μm, with a slightly apiculate base,
unicellular, intercalary, usually in chains, subglobose, oval to pyriform, 5–9 × 4–6 μm,
Specimens examined: USA. NEVADA: From human toe nail, D.A. Sutton (CBS H-20782,
HOLOTYPE; UTHSC 04–7, CBS 131712, FMR 11070, ex-type cultures). USA. MINNESOTA: From human
Phialemoniopsis pluriloculosa differs from the other species of the genus by the
II).
Emended description.
Colonies spreading, flat, velvety, plane, or slightly floccose, white, yellow-gray, ochraceous
Phialides often inflated at base, with or without barely visible collarettes. Adelophialides
slimy heads or in long chains, with or without connectives at both ends. Chlamydospores
Sordariales).
base and more or less conspicuous collarettes, and conidia arranged in long chains that may
Phialemonium atrogriseum (Panas.) D. García, H. Perdomo, Gené, Cano & Guarro, comb.
(1971).
for growth was at 25 C. Conidiophores absent or poorly differentiated, often single lateral
phialides born directly from aerial hyphae , or consisting of whorls of 2–4 phialides on top of
lateral supporting cells. Phialides flask-shaped with more or less inflated basal part and a
long, tapering neck bearing a barely visible collarete, 8–20 × 2–3.5 µm wide at base, mostly
isolates. Adelophialides may be also present, 7–11 × 1.5–2 µm. Conidia 1-celled, ellipsoidal
or obovoid, 3–5 × 2–3 µm, with an apiculate and minutely truncate base, subhyaline to
grayish brown, smooth-walled, in slimy heads and in long chains, without connectives.
Specimens examined: UKRAINE. Kharkiv: From noodles, V.T. Panasenko (CBS 604.67, ex-type
culture). GERMANY. München; From human urine, A. Haas (CBS 774.97). USA. DELAWARE: Dover. From
a lower lobe lung, A.A. Padhye (CBS 306.85). USA. PENNSYLVANIA: From BAL aplastic anemia, D.A.
chains by its slightly pigmented conidia without connectives. Although P. atrogriseum forms
conidia similar to those of P. obovatum, in the latter they are not pigmented and are arranged
only in slimy heads. Also, P. obovatum produces chlamydospores and P. atrogriseum does
not.
Phialemonium globosum D. García, H. Perdomo, Gené, Cano & Guarro, sp. nov. FIG. 8H–L
MycoBank MB563880
Colonies on OA and PDA at 25 C attained up to 30 mm diam after 14 d, flat, velvety to
powdery, sometimes fasciculate in the center, white. The fungus grew slowly at 37 C (up to
Conidiophores absent or poorly differentiated, often single lateral phialides borne directly
from aerial hyphae, occasionally 1–2 phialides borne on top of a lateral supporting cell.
Phialides terminal or lateral, sometimes sympodially proliferating from the supporting cell
without basal septa, cylindrical or with a slightly swollen basal part, sometimes slightly
constricted at midpoint, and a long tapering neck, (7–)9–24(–36) × 1.5–3 µm near the base, 1–
4–6 × 3.5–5 µm, with distinct connectives on both ends, hyaline, smooth-walled, forming
Specimen examined: USA. ILLINOIS: From human sinus, D.A. Sutton (CBS H-20783, HOLOTYPE;
and Phialemonium inflatum (see below). They all share phialides with a more or less swollen
basal portion and form long conidial chains with connectives at both ends. However, these
three species can be distinguished by their conidial morphology: P. globosum has globose
conidia up to 5 µm diam, the anamorph of C. sulfurea has ovoid conidia up to 3 µm diam, and
Phialemonium inflatum (Burnside) D. García, H. Perdomo, Gené, Cano & Guarro, comb.
MycoBank MB563881
≡ Taifanglania inflata (Burnside) Z.Q. Liang, Y.F. Han & H.L. Chu, Fungal Diversity 34:72 (2009)
occasionally 2–3 phialides borne atop lateral supporting cells. Phialides lateral or terminal,
flask-shaped, with a swollen basal part and tapering abruptly into a long neck, 7–15(–19) ×
(1.5–)2–3 µm, hyaline, smooth-walled. Conidia one-celled, citriform, 4–5 × 2–3 µm, with
distinct connectives on both ends, hyaline, smooth- to slightly rough-walled, and arranged in
Specimens examined: USA. WASHINGTON: From Apis mellifera, J.A. Stevenson (CBS 259.39, ex-
type culture). As Taifanglania inflata: RUSSIA. Ust-Omchug. From cabbage field soil, T. Ito and T. Yokoyama
(NBRC 31965).
The key morphological features, based on OA cultures, useful for distinguishing the
Phialemonium species included in the present study, are summarized (TABLE III).
Lecythophora cateniformis H. Perdomo, D. García, Gené, Cano & Guarro, sp. nov. FIG.
9A–I
MycoBank MB563407
center and white at the periphery; reverse brownish gray (8F2). On PDA at 25 C, the colonies
reaching 35 mm diam after 14 d, flat, velvety, moist, orange-white (6A2); reverse pale yellow
pyriform, 4–7(–9) × 4–5(–6) μm, pale brown to brown, smooth and slightly thick-walled.
Teleomorph unknown.
Specimen examined: USA. ALABAMA: From canine bone marrow aspirate, D.A. Sutton (CBS H-
20779, HOLOTYPE; UTHSC 01-1644, CBS 131709, FMR 10743, ex-type cultures).
hyaline or faintly brown chlamydospores and its conidia are ellipsoidal to cylindrical up to 2
μm wide.
Lecythophora luteorubra H. Perdomo, D. García, Gené, Cano & Guarro, sp. nov. FIG. 9J–P
MycoBank MB563408
orange (6B8); reverse orange (6A7). Colonies on PDA at 25 C attained 25 mm diam after 14
d, downy, orange (6B8); reverse orange (6B7). Diffusible pigment orange-red to brownish red
(10D8) was produced on both media at 25 C, but it is absent at 30 C. The fungus grew slowly
cylindrical or ampulliform, 5–10 × 1–3 μm, with distinct cylindrical collarettes, hyaline,
hyphae, 1–1.5 × 1 μm. Conidia one-celled, ellipsoidal or cylindrical, 3–6(–7) × 1–2 μm, with
Etymology: From the Latin luteorubra, referred to the orange-red diffusible pigment.
Specimen examined: USA. MASSACHUSETTS: From leg wound, D.A. Sutton (CBS H-20780,
anamorph of Coniochaeta ligniaria (Grev.) Massee. However, L. hoffmannii does not produce
diffusible pigment, and its conidia are slightly smaller (3.5–5 × 1.2–2 μm) and often
somewhat more curved than those of L. luteorubra. In the anamorph of C. ligniaria, the
conidia often are curved; the adelophialides are predominant; although some cylindrical
phialides may be present, they are never ampulliform; and the teleomorph is usually present.
The key morphological features useful to distinguish current Lecythophora species are
DISCUSSION
The genus Phialemonium was proposed to accommodate three species with morphological
features intermediate between Acremonium and Phialophora (Gams and McGinnis 1983).
Yaguchi et al. (2006) and Perdomo et al. (2011b) demonstrated that, as originally conceived,
the genus was polyphyletic. They suggested that P. curvatum should be accommodated in
different genus. Based on combined morphological and additional molecular data, we propose
the new anamorphic genus Phialemoniopsis to accommodate P. curvatum and three additional
species, Phialemoniopsis cornearis and P. pluriloculosa proposed here as new and P. ocularis
Other anamorphic genera that have the same conidial ontogeny and develop
textura angularis and by the production of fusiform or falcate conidia. Molecular data placed
the members of Chaetomella within the Leotiomycetes (Helotiales) (Rossman et al. 2004).
fusispora (P. Karst.) Seifert. Their conidiomata are usually cylindrical, sessile and non-setose,
with a wall of textura porrecta (Sutton 1980, Seifert 1985). The taxonomical placement of
Cornucopiella is still uncertain among ascomycetes. Cultures of the first species are not
available for study; however, we examined and sequenced the D1/D2 and ITS regions of a
reference strain of C. fusispora (CBS 113544). That strain did not sporulate in any agar media
tested. The BLAST query of the ITS and D1-D2 of it showed similarities below 91% with
respectively.
white, cream or reddish colonies and pycnidium-like conidiomata with walls of textura
epidermoidea (Damm et al. 2010). Sporodochial conidiomata is not present in this genus. The
species of Collophora are known only as agents of wood necroses of Prunus trees (Damm et
al. 2010).
Sarcopodium they are brown pinkish or purple, with brown setae (Sutton 1981, Domsch et al.
2007), and in Volutella they are white to pale with hyaline setae around the margin (Maud and
Duke 1928, Gräfenhan et al. 2011). Most species of these two genera are saprotrophic or
pathogenic on different plants, but two of them, V. cinerescens and S. oculorum, have been
described as human opportunists of ocular infections (de Hoog et al. 2000, Guarro et al.
2002). Our molecular analysis shows that type species of Sarcopodium (S. circinatum) and
Volutella (V. ciliata) are distantly related to Phialemoniopsis in the Hypocreales. The isolates
of the respective opportunistic species wrongly assigned to those genera were clearly related
to P. curvata (FIG. 1). That is the case of V. cinerescens CBS 832.71, isolated from a case of
endophthalmitis, whose affinity to P. curvata was suggested by Proia et al. (2004). That
isolate, as well as other clinical isolates included in the Proia et al. (2004) study, developed
the typical sporodochium-like conidiomata described for P. curvata in the present study. It is
noteworthy, however, that such structures were not observed in the type strain and in two
other reference strains of P. curvata in any of the media tested in this study. These are
relatively old strains and we conjectured that they have degenerated, thereby losing the ability
to produce conidiomata.
The description of S. oculorum was based on two isolates obtained from a corneal
ulcer of a human patient (Guarro et al. 2002). These isolates were initially identified as
media tested, a character shared with P. curvata. Therefore, the presence of conidiomata and
the phylogenetic relationships with the latter fungus support inclusion of S. oculorum in
In the D1/D2 analysis (FIG. 1), Phialemoniopsis was phylogenetically related to some
species of the genera Linocarpon (L. livistonae) and Neolinocarpon (N. globosicarpum and N.
enshiense). Those genera show anamorphs producing phialides with collarettes similar to
those present in Phialemoniopsis (Hyde et al. 1998, Bahl 2006). However, Huhndorf and
Miller (2011) demonstrated that Linocarpon is polyphyletic and the species of that genus and
of Neolinocarpon, particularly related with Phialemoniopsis, were not associated with any
known family. We could not include Neolinocarpon species in our combined analysis of
DNA phylogenies because strains of members of that genus are not available in any public
culture collection
and Cephalotheca, and let us enlarge the circumscription of the genus Phialemonium to
include species producing conidia in slimy heads or in chains. We found a close phylogenetic
inflata, Acremonium atrogriseum (FIGS. 1, 4). The genus Taifanglania was erected by Liang
et. al. (2009) to accommodate thermotolerant fungi, as well as some Paecilomyces species
showing single phialides on undifferentiated hyphae like Paecilomyces inflatus. The ex-type
strain of P. inflatus, isolated from a worker bee in USA, and the strain used by Liang et al.
(2009) to propose T. inflata, isolated from soil in Russia, were studied. They proved to be
conspecific and related to the type species of Phialemonium. It is worth mentioning that T.
inflata is the only Taifanglania species that is not thermotolerant. According to our
members of Taifanglania are placed within the Chaetomiaceae (Liang et al. 2009). These
inflatum is closely related to C. sulfurea and P. globosum (FIG. 4). Morphologically, they all
develop discrete phialides with more or less swollen bases, usually without visible collarettes,
and produce long persistent chains of conidia with connectives (Chester 1935, Onions and
Barron 1967, Liang et al. 2009). Differences among these species are summarized (TABLE
III). Our polyphasic taxonomic approach also supports the accommodation of A. atrogriseum
in the genus Phialemonium. This rare opportunistic fungus differs morphologically from the
other Phialemonium species by the lack of chlamydospores and by producing grayish brown
Sagenomella W. Gams, Phialosimplex Sigler, Deanna A. Sutton, Gibas, Summerb. & Iwen,
Monocillium S.B. Saksena and Septofusidium W. Gams. However, all these fungi produce
only catenulate conidia. These genera are phylogenetically unrelated to Phialemonium; the
first two genera belong to the Trichocomaceae (Eurotiales), while Monocillium is placed in
the Niessliaceae and Septofusidium in the Nectriaceae, both latter within Hypocreales
Perdomo et al. (2011b) showed that Lecythophora and Coniochaeta constituted a well
defined group with several unidentified clinical isolates. Here, we have described two new
species closely related to L. hoffmannii (FIG. 3); however, other putative new species (i.e.
Lecythophora sp. 2–4) were not formally proposed because they could not be clearly
distinguished morphologically from the currently accepted Lecythophora species. The novel
species, L. luteorubra and L. cateniformis, are distinguished from each other mainly by the
respectively (TABLE IV). It is important to note that descriptions of those two species are
based on single isolate each. Therefore, phenotypic identification characters are susceptible to
ACKNOWLEDGMENTS
The authors are indebted to the curators of the Centraalbureau voor Schimmelcultures (Utrecht, the Netherlands),
BCCM™/IHEM Biomedical Fungi and Yeasts Collection (Brussels, Belgium) and NITE Biological Resource
Center (Chiba, Japan) for supplying many of the strains used in the study. This work was supported by the
Spanish Ministerio de Ciencia e Innovación grants CGL 2009-08698/BOS and CGL 2011-27185/BOS.
LITERATURE CITED
Bahl J. 2006. Molecular evolution of three morphologically similar families in the Xylariomycetidae
lineage for Stachybotrys chartarum, the indoor air fungus. Mycol Res 108:864–872.
Cano J, Sagués M, Barrio E, Vidal P, Castañeda RF, Gené J, Guarro J. 2002. Molecular taxonomy of
Aphanoascus and description of two new species from soil. Stud Mycol 47:153–164.
Chesters CGC. 1935. Studies on British Pyrenomycetes. The life histories of three species of Cephalotheca
Damm U, Fourie PH, Crous PW. 2010. Coniochaeta (Lecythophora), Collophora gen. nov. and Phaeomoniella
de Hoog GS, Guarro J, Gené J, Figueras MJ. 2000. Atlas of clinical fungi. 2nd ed. Utrecht, the Netherlands :
Domsch KH, Gams W, Anderson T. 2007. Compendium of soil fungi. 2nd ed. München, Germany: IHW
Verlag. 672 p.
Drees M, Wickes BL, Gupta M, Hadley S. 2007. Lecythophora mutabilis prosthetic valve endocarditis in a
Verlag. 262 p.
———, McGinnis M. 1983. Phialemonium, a new anamorph genus intermediate between Phialophora and
Gilgado F, Cano J, Gené J, Guarro J. 2005. Molecular phylogeny of the Pseudallescheria boydii species
typification of nectriaceous fungi in Cosmospora, Acremonium, Fusarium, Stilbella and Volutella. Stud Mycol
68:79–13.
Guarro J, Höfling-Lima A, Gené J, de Freitas D, Godoy P, Zorat-Yu M, Zaror L, Fischman O. 2002. Corneal
ulcer caused by the new fungal species Sarcopodium oculorum. J Clin Microbiol 40:3071–3075.
———, Nucci M, Akiti T, Gené J, Cano J, Barreiro MDC, Aguilar C. 1999. Phialemonium fungemia: two
Huelsenbeck JP, Ronquist F. 2001. MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:754–755.
Huhndorf SM, Miller AN. 2011. A molecular re-appraisal of taxa in the Sordariomycetidae and a new species of
Hyde K, Taylor JE, Fröhlich J. 1998. Fungi from palms XXXIV. The genus Neolinocarpon with five new
King D, Pasarell L, Dixon DM, McGinnis MR, Merz WG. 1993. A phaeohyphomycotic cyst and peritonitis
caused by Phialemonium species and a reevaluation of its taxonomy. J Clin Microbiol 31:1804–1810.
Kornerup A, Wanscher JH. 1978. Methuen handbook of color. 3rd ed. London: Erye Methuen. 252 p.
Liang ZQ, Han YF, Chu HL, Fox RTV. 2009. Studies on the genus Paecilomyces in China V. Taifanglania gen.
Luangsa-ard JJ, Hywel-Jones NL, Samson RA. 2004. The polyphyletic nature of Paecilomyces sensu lato based
184.
Onions AHS, Barron GL. 1967. Monophialidic species of Paecilomyces. Mycol Pap 107:1–25.
Perdomo H, Sutton DA, García D, Fothergill AW, Cano J, Gené J, Summerbell RC, Rinaldi MG, Guarro J.
2011a. Spectrum of clinically relevant Acremonium species in the United States. J Clin Microbiol 49:243–256.
Perdomo H, Sutton DA, García D, Fothergill AW, Gené J, Cano J, Summerbell RC, Rinaldi MG, Guarro J.
2011b. Molecular and phenotypic characterization of Phialemonium and Lecythophora isolates from clinical
Proia LA, Hayden MK, Kammeyer P, Ortiz J, Sutton DA, Clark T, Schroers H-J, Summerbell RC. 2004.
Phialemonium: an emerging mold pathogen that caused four cases of hemodialysis-associated endovascular
Rivero M, Hidalgo A, Alastruey-Izquiedo A, Cía M, Torroba L, Rodríguez-Tudela JL. 2009. Infections due to
Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed models.
Bioinformatics 19:1572–1574.
Rossman AY, Aime MC, Farr DF, Castlebury LA, Peterson KR, Leahy R. 2004. The coelomycetous genera
Chaetomella and Pilidium represent a newly discovered lineage of inoperculate discomycetes. Mycol Prog
3:275–290.
Sakaeyama S, Sano A, MurataY, Kamei K, Nishimura K, Hatai K. 2007. Lecythophora hoffmannii isolated from
Seifert K. 1985. Notes on several apocryphal genera of synnematal hyphomycetes. Trans Br Mycol Soc 85:123–
133.
———, Morgan-Jones G, Gams W, Kendrick B. 2011. The genera of Hyphomycetes. Utrecht, the Netherlands:
Sigler L, Sutton DA, Connie FC, Summerbell RC, Noel RK, Iwen PC. 2010. Phialosimplex, a new anamorphic
genus associated with infections in dogs and having phylogenetic affinity to the Trichocomaceae. Med Mycol
48:334–345.
Schwammenthal Y, Feinberg MS, Siegman-Igra Y, Bash E, Polacheck I, Zelazny A, Howard SJ, Cibotaro P,
Shovman O, Keller N. 2005. An outbreak of Phialemonium infective endocarditis liked to intracavernous penile
Suh S-O, Blackwell M. 1999. Molecular phylogeny of the cleistothecial fungi placed in Cephalothecaceae and
Summerbell RC, Gueidan C, Schroers H-J, de Hoog GS, Starink M, Rosete A, Guarro J, Scott JA. 2011.
Acremonium phylogenetic overview and revision of Gliomastix, Sarocladium and Trichothecium. Stud Mycol
68:139–162.
Sutton BC. 1980. The Coelomycetes: fungi imperfecti with pycnidia, acervuli and stromamata. Kew, UK:
———. 1981. Sarcopodium and its synonyms. Trans Brit mycol Soc 76:97–102.
Sutton DA, Wickes BL, Thompson EH, Rinaldi MG, Roland RM, Libal MC, Russel K, Gordon S. 2008.
Pulmonary Phialemonium curvatum phaeohyphomycosis in a standard poodle dog. Med Mycol 46:355–359.
Swofford DL. 2002. PAUP*: phylogenetic analysis using parsimony (*and other methods). 4.0b10. Sunderland,
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary
genetics analysis using maximum likelihood, evolutionary distance and maximum parsimony methods. Mol Biol
Evol 28:2731–2739.
Taniguchi Y, TaketanT, Moriyama H, Moriki S, Nishimura K, Sato E, Notsu Y, Higuchi T, Sugitani Y, Yasuda
K, Nagai A, Yamaguchi S, Shibata H, Masuda J. 2009. Septic shock induced by Lecythophora mutabilis in a
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The Clustal X Windows interface:
flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–
4882.
van Heerden A, Willem H van Z, Cruywagen CW, Mouton M, Botha A. 2011. The lignicolous fungus
Coniochaeta pulveracea and its interactions with syntrophic yeasts from the woody phylloplane. Microb Ecol
62:609–619.
Voigt K, Wöstemeyer J. 2000. Reliable amplification of actin genes facilitates deep-level phylogeny. Microbiol
Res 155:179–195.
———, Görke C, Begerow D. 2002. The Lecythophora-Coniochaeta complex II. Molecular studies based on
Pitlik SD, Rinaldi MG, Thompson E, Sutton DA. 2006. Isolated endogenous endophthalmitis due to a
44:253–259.
Yaguchi T, Sano A, Yarita K, Suh MK, Nishimura K, Udagawa S. 2006. A new Cephalotheca isolate from
LEGENDS
FIG. 1. Maximum likelihood (ML) tree obtained from D1/D2 domains of the 28S rDNA sequences of isolates
(TABLE I). Numbers on the branches are bootstrap ML values above 55%, followed by Bayesian posterior
probabilities (Bpp) above 0.56. Branch lengths are proportional to distance. Sequences not generated in this
study and obtained from the GenBank database are indicated in parentheses. Type strains of the different species
FIG. 2. One of the two most parsimonious trees inferred from heuristic search of a combined alignment of the
domains D1/D2, ITS, ACT and TUB of the isolates included in clade I (FIG. 1). Bootstrap support values above
70% are indicated at the nodes. Lecythophora lignicola (CBS 267.33) was chosen as outgroup. GenBank
accession numbers for sequences included in this analysis are listed (TABLE I). T Type strain. New taxa are in
boldface.
FIG. 3. One of the two most parsimonious trees inferred from heuristic search of a combined alignment of the
domains D1/D2, ITS, ACT and TUB of the isolates included in the clade III (FIG. 1). Bootstrap support values
above 70% are indicated at the nodes. Phialemonium obovatum (CBS 279.76) was chosen as outgroup. GenBank
accession numbers for sequences included in this analysis are provided (TABLE I). The news species are
FIG. 4. The most parsimonious tree inferred from heuristic search of a combined alignment of the domains
D1/D2, ITS, ACT and TUB of the isolates included in clade IV (FIG 1). Bootstrap support values above 70% are
indicated at the nodes. Lecythophora lignicola (CBS 267.33) was chosen as outgroup. GenBank accession
numbers for sequences included in this analysis are provided (TABLE I). T Type strain. New taxa are in boldface.
FIG. 5. Phialemoniopsis cornearis, UTHSC 06-1820T. A. Colony surface on OA after 14 d at 25 C. B. Colony
surface on OA after 6 mo incubation at 25 C with conidiomata. C. Conidiophore from aerial mycelium. D–L.
Phialides, adelophialides (arrows) and conidia from aerial mycelium. M. Chlamydospore. N, O. Sporodochium-
like conidiomata. P. Conidiophores lining the inner wall of sporodochium-like conidiomata. Q, R. Detail of
with wall of textura intricata. U, V. Detail of branched conidiophores from pycnidium-like conidiomata and
conidia. Bars: C–I, M, Q = 10 μm; J–L, R, V = 5 μm; N, O, S = 100 μm; T = 50 μm; P, U = 20 μm.
FIG. 6. Phialemoniopsis ocularis, IHEM 19077T. A–E. Phialides, adelophialides and conidia from the aerial
mycelium. F, G. Pycnidium-like conidiomata. H–J. Conidiophores lining the inner wall of a sporodochium-like
conidioma and conidia. K, L. Chlamydospores. Conidiophores from a pycnidium-like conidiomata and conidia.
D–F. Phialides, adelophialides (arrow) and conidia from the aerial mycelium. G, H, J. Multilocular pycnidium-
Conidiophores and conidia from a pycnidium-like conidiomata. Bars: G, H, J = 100 μm; C, D, I, K = 10 μm; E,
F, L = 5 μm.
FIG. 8. A–G. Phialemonium atrogriseum, CBS 604.67T. A, B. Colony surface and reverse on OA after 14 d at
25 C. C–G. Phialides, some with a visible collarette (arrow) and conidia. H–L. Phialemonium globosum,
UTHSC 03-3661T. H. Colony surface on OA after 14 d at 25 C. I–L phialides and conidia in chain. M–Q.
conidia. O. Conidia in chains. P, Q. Phialide and conidia. Bars: C, I, J, N = 10 μm; D–F, G, K, L, O–Q = 5
μm.
FIG. 9. A–I. Lecythophora cateniformis, UTHSC 01-1644T. A, B. Colony surface on PDA and OA respectively
Lecythophora luteorubra, UTHSC 01-20-1T. J, K. Colony surface and reverse on PDA after 14 d at 25 C. L.
Colony surface and diffusible pigment on PDA after 2 mo incubation at 25 C. M. Phialides and conidia in slimy
heads. N, O. Phialides without basal septum. P. Conidia. Bars: G–I, M–O = 10 μm; C–F, P = 5 μm.
FOOTNOTES
Submitted 19 Apr 2012; accepted for publication 11 Sep 2012.
1
Corresponding author. E-mail: josepa.gene@urv.cat
TABLE I. Phialemonium and Lecythophora clinical isolates and type or reference strains of related species included in the study
P. ocularis Whitish to brownish Solitary, superficial, hyaline to subhyaline. Solitary, superficial, globose to subglobose, dark Chlamydospores present
gray Conidia subhyaline, ellipsoid, navicular or brown to black, unilocular, textura intricata and
slightly allantoid epidermoidea. Conidia hyaline, ellipsoidal to
lightly allantoid, slightly apiculate at the base
P. pluriloculosa Yellowish white, olive- Solitary, superficial, multilocular, Solitary or in groups, superficial or semi- Chlamydospores present
brown to black subhyaline to brown. Conidia hyaline, immersed, globose to subglobose, dark brown to
ellipsoidal black, multilocular, textura epidermoidea.
Conidia hyaline, ellipsoidal, slightly apiculate at
the base
TABLE III. Distinctive morphological features of Phialemonium species based on OA cultures at 25 C after 14 d
Species Colonies Conidia Other distinctive features
P. atrogriseum Pale ocher-brown to Subhyaline to dark gray, ellipsoidal or Sometimes, phialides with a short pale
brownish black obovoid with an apiculate and minutely collarette. Ascomata and chlamydospores
truncate base, in slimy heads and in chains absent
without connectives
P. globosum White Hyaline, globose to subglobose, connective Chlamydospores and ascomata absent
on both ends, in chains
P. inflatum White to pale yellow Hyaline, citriform, connective on both Chlamydospores and ascomata absent
ends, in chains
P. obovatum Pale ochraceous to Hyaline, obovoid with a truncate base, in Chlamydospores hyaline, oval and thin-
greenish slimy heads walled. Some isolates develop globose and
dark structures suggestive of immature
ascomata
Phialemonium White to brownish Hyaline, cylindrical, in slimy heads Chlamydospores hyaline, ellipsoidal, thick-
anamorph of walled. Ascomata present, covered with
Cephalotheca sulfurous to glabrous hyphae; ascospora
foveolata elliptic-ovate, foveolate-walled
Phialemonium White to pale yellow Hyaline, ovoid, connective on both ends, in Chlamydospores absent. Ascomata present,
anamorph of chains covered with light yellow to brown hyphal
Cephalotheca sulfurea hairs; ascospores broadly reniform, smooth-
walled
TABLE IV. Distinctive morphological features of Lecythophora species, based on OA cultures at 25 C after 14 d
Species Colonies Conidia Chlamydospores Other distinctive features
L. cateniformis Brownish gray Obovoid to ellipsoidal, with a Pale brown to brown, subglobose, Adelophialides are
protruding and slightly truncate oval to pyriform, solitary or in predominating
base short chains
L. decumbens Olivaceous to Ellipsoidal to cylindrical, often Absent Presence of conidia swollen up
brown somewhat curved to 13 × 3–4.5 μm
L. fasciculata Gray-brown to Ovoid to ellipsoidal, often Absent Adelophialides are
black somewhat curved predominating
L. hoffmannii Orange to salmon Ellipsoidal to cylindrical, often Absent More common short
somewhat curved adelophialides than ventricose
phialides
L. luteorubra Orange Ellipsoidal to cylindrical, slightly Absent Colonies producing a diffusible
apiculate at the base pigment orange-red to
brownish-red
L. luteoviridis Olivaceous to Ellipsoidal to cylindrical, often Hyaline or faintly brown, globose Darkening of the colonies due
brown somewhat curved to oblong or pear-shaped, singly to development of pigmented
or short chains chlamydospores
L. lignicola Brown Ovoid to ellipsoidal Absent Discrete ventricose phialides
abundant
L. mutabilis Brown Ellipsoidal, rarely curved Olivaceous-brown, ovoid to Discrete ventricose phialides
ellipsoidal, more rarely globose, abundant
singly
CBS 490.82T Phialemonium curvatum
CBS 491.82
UTHSC 06-4324
UTHSC R-3447
99/1 UTHSC R-3448
CBS 832.71
UTHSC 08-2292
99/1 UTHSC 04-956
UTHSC 09-3589
UTHSC 06-1465
Clade I
UTHSC 06-1820 Incertae sedis
UTHSC 09-2358 Incertae sedis
87/0,9
UTHSC 07-3736
IHEM 19077T Sarcopodium oculorum
FMR 7190
UTHSC 05-2527
UTHSC 04-7
Linocarpon livistonae (DQ810206)
94/0,98 Neolinocarpon globosicarpum (DQ810224)
Neolinocarpon enshiense (DQ810221)
UTHSC 06–1820 T
100 100 Phialemoniopsis cornearis
UTHSC 06–1465
UTHSC 04–7 T
96 Phialemoniopsis pluriloculosa
UTHSC 09–3589
UTHSC 06–4324
83 CBS 491.82
UTHSC 08–2292
98 UTHSC 04–956
1
D1-D2, ITS, ACT, TUB
2173 bp CBS 205.38 T Lecythophora fasciculata
Tree length= 514 77
CI=0.65 100 CBS 267.33 T Lecythophora lignicola
HI=0.35
RI=0.55< 98 CBS 206.38 T Lecythophora luteoviridis
UTHSC 01–1399
UTHSC 06–733
UTHSC 03–2653
100
UTHSC 03–986
CBS 774.97
100
Phialemonium atrogriseum
CBS 306.85
90 CBS 604.67 T Acremonium atrogriseum
10