International Journal of Systematic and Evolutionary Microbiology (2014), 64, 3856–3861
DOI 10.1099/ijs.0.068403-0
Wickerhamiella allomyrinae f.a., sp. nov., a yeast
species isolated from the gut of the rhinoceros
beetle Allomyrina dichotoma
Yong-Cheng Ren, Yun Wang, Liang Chen, Tao Ke and Feng-Li Hui
Correspondence
Feng-Li Hui
[email protected]
School of Life Science and Technology, Nanyang Normal University, Nanyang 473061, PR China
Two strains representing Wickerhamiella allomyrinae f.a., sp. nov. were isolated from the gut of
Allomyrina dichotoma (Coleoptera: Scarabeidae) collected from the Baotianman National Nature
Reserve, Nanyan, Henan Province, China. Sequence analyses of the D1/D2 domains of the LSU
rRNA gene revealed that this novel species was located in the Wickerhamiella clade
(Saccharomycetes, Saccharomycetales), with three described species of the genus Candida,
namely Candida musiphila, Candida spandovensis and Candida sergipensis, as the most closely
related species. The novel species differed from these three species by 9.3–9.8 % sequence
divergence (35–45 nt substitutions) in the D1/D2 sequences. The species could also be
distinguished from the closely related species, C. musiphila, C. spandovensis and C. sergipensis,
by growth on vitamin-free medium and at 37 6C. The type strain is Wickerhamiella allomyrinae sp.
nov. NYNU 13920T (5CICC 33031T5CBS 13167T).
The ascomycetous yeast genus Wickerhamiella is closely
related to the genera Spencermartinsiella, Starmerella,
Sugiyamaella, Trichomonascus, Yarrowia and Zygoascus on
the basis of concatenated gene sequences for the nearly
entire LSU rRNA, SSU rRNA, translation elongation
factor-1a (EF-1a), and RNA polymerase II subunits 1
(RPB1) and 2 (RPB2) (Kurtzman & Robnett, 2013). The
type strain of the genus was initially described as Torulopsis
domercqiae (as Torulopsis domercqii) by van der Walt & van
Kerken (1960). Van der Walt & Liebenberg established
the new genus Wickerhamiella in 1973, and transferred T.
domercqiae to this genus as Wickerhamiella domercqiae
after examining two additional strains and observing
conjugation and ascospore formation in two of the three
available strains (van der Walt & Liebenberg, 1973). In the
fifth edition of The Yeasts, A Taxonomic Study, the genus
Wickerhamiella accommodated five species: Wickerhamiella
australiensis, Wickerhamiella cacticola, W. domercqiae, Wickerhamiella lipophila and Wickerhamiella occidentalis (Lachance
& Kurtzman, 2011). There are more than 17 Candida species
that are currently members of the genus Wickerhamiella on
the basis of sequence-based phylogenetic analyses (Lachance
& Kurtzman, 2011; Badotti et al., 2013). Subsequently, several
additional members of the genus Wickerhamiella have been
proposed, such as Wickerhamiella pagnoccae (Barbosa et al.,
Abbreviation: ITS, internal transcribed spacer.
The GenBank/EMBL/DDBJ accession numbers for the sequences
of the D1/D2 domains of the LSU rRNA gene and the ITS regions
of Wickerhamiella allomyrinae NYNU 13920T are KJ152751 and
KJ152752, respectively.
3856
2012), Wickerhamiella dulcicola and Wickerhamiella cachassae
(Badotti et al., 2013), Wickerhamiella slavikovae and Wickerhamiella goesii (Hagler et al., 2013), and Wickerhamiella
kiyanii and Wickerhamiella fructicola (Dayo-Owoyemi et al.,
2014). Given that yeasts of the genus Wickerhamiella are
physiologically similar in their utilization of carbon and
nitrogen compounds, separating them is difficult on the basis
of phenotypic characteristics. Therefore, species identification
should be based on rRNA gene sequence comparisons.
During a study of yeasts associated with insects, we isolated
a large number of yeasts from the digestive tract of insects
as well as from related substrates, including rotting wood,
frass and galleries (Chen et al., 2013; Hui et al., 2013). The
majority of the yeasts belonged to several major clades in
the subphylum Saccharomycotina; some of these species
have been identified as novel species in earlier papers (Hui
et al., 2012, 2013a, 2013b; Chen et al., 2013). Amongst the
insect associates, we focused on two strains of an asexual
ascomycetous yeast species from the gut of the rhinoceros
beetle Allomyrina dichotoma in China. Sequence analysis of
the D1/D2 domains of the LSU rRNA gene revealed
that the aforementioned strains represent an undescribed
anamorphic yeast species belonging to the Wickerhamiella
clade. In this paper, we describe this novel species as
Wickerhamiella allomyrinae f.a., sp. nov.
The strains belonging to the proposed novel species,
NYNU 13915 and NYNU13920T, were isolated from the
gut of two individuals of A. dichotoma in September 2013.
Adult insects were collected from the Baotianman National
Nature Reserve near Nanyang (approximate coordinates:
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Printed in Great Britain
Wickerhamiella allomyrinae f.a., sp. nov.
33u 279 N and 111u 489 E), which has a typical transitional
climate from the northern subtropical zone to the warm
temperate zone in central China.
The methods for yeast isolation were detailed by Nguyen
et al. (2006) and Urbina et al. (2013). The insects were
usually placed in Petri dishes for 1–3 days without food
prior to dissection. Withholding food helps to eliminate
some contaminating organisms that might be isolated from
the gut. Each insect individual was surface disinfected by
washing in 70 % ethanol (5 min), 5 % bleach (5 min) and
sterile water (10 min) prior to dissection. The aseptically
removed gut contents and saline wash solution were plated
separately on acidified yeast extract-malt extract (YM) agar
(0.3 % yeast extract, 0.3 % malt extract, 0.5 % peptone, 1 %
glucose, 2 % plain agar, adjusted to pH 3.5 with HCl), and
incubated at 25 uC for 3–4 days. Single yeast colonies were
purified at least twice, and stored in 15 % glycerol at
280 uC and/or on YM agar at 4 uC.
The morphological observations and metabolic tests that
constitute the standard yeast description were performed
according to established methods (Yarrow, 1998; Kurtzman
et al., 2011). Assimilation tests for carbon and nitrogen
sources were performed in liquid media. Starved inocula
were used in nitrogen and vitamin assimilation tests. Strains
were examined for ascosporulation on the following agar
media incubated at 15 uC and 25 uC for 1–4 weeks: YM
agar, 5 % malt extract agar, corn meal agar and YCBAS agar
(1.1 % yeast carbon base, 0.01 % ammonium sulphate and
1.8 % agar).
Genomic DNA was extracted using an Ezup Column Yeast
Genomic DNA Purification kit according to the manufacturer’s protocol (Sangon Biotech). The D1/D2 domains
of the LSU rRNA gene and ITS regions were amplified by
PCR, and sequenced using primers NL1 and NL4 (Kurtzman
& Robnett, 1998) and ITS1 and ITS4 (White et al., 1990),
respectively. Both DNA strands were sequenced, and the
reactions were carried out using a Dye Terminator cycle
sequencing kit (Applied Biosystems).
The sequences were compared in a pairwise manner using a
search (Altschul et al., 1997), and aligned with the
sequences of related species retrieved from GenBank using
the multiple alignment program CLUSTAL X version 1.81
(Thompson et al., 1997). A phylogenetic tree based on LSU
D1/D2 sequences was reconstructed using the neighbourjoining method in MEGA 5.0 (Tamura et al., 2011).
Evolutionary distance data were calculated from Kimura’s
two-parameter model (Kimura, 1980) in the neighbourjoining analyses. All sites containing gaps in the alignment
were excluded. Dipodascus magnusii NRRL Y-17563T and
Starmerella bombicola NRRL Y-17069T were used as outgroups.
Confidence levels of the clades were estimated from
bootstrap analysis (1000 replicates) (Felsenstein, 1985),
and only values above 50 % were recorded on the resulting
tree. Reference sequences were retrieved from GenBank
under the accession numbers indicated on the tree
(Fig. 1).
BLAST
http://ijs.sgmjournals.org
Sequence comparison and species delineation
The two strains of W. allomyrinae sp. nov. were found to
share identical sequences in both D1/D2 and ITS regions.
Sequence analyses of the D1/D2 domains of the LSU rRNA
gene revealed that this novel species was closely related to
species in the Wickerhamiella clade (Saccharomycetes,
Saccharomycetales). In terms of pairwise sequence similarity, the close matches to W. allomyrinae sp. nov. were
Candida musiphila, Candida spandovensis and Candida
sergipensis. The D1/D2 sequences of the novel species
showed a sequence divergence of 9.3 % (35 substitutions
and 17 gaps over 548 bases) from the closest relative C.
musiphila. The novel species also differed from its other
two close relatives, C. spandovensis and C. sergipensis, by
sequence divergences of 9.4 % and 9.8 %, respectively. For
the ITS region, this novel species differed by sequence
divergence of 13.3 % and 21.4 % from C. musiphila and C.
spandovensis, respectively. However, pairwise sequence
analysis with C. sergipensis could not be performed because
its ITS sequences are not currently available from either the
NCBI GenBank database or the CBS database.
A phylogenetic analysis based on the D1/D2 domains of the
LSU rRNA gene sequences indicated that W. allomyrinae
sp. nov. forms a subclade with C. spandovensis and C.
sergipensis (Fig. 1). The bootstrap support for this subclade
was relatively low (86 %; Fig. 1). The high degree of
sequence divergence among the described species of the
subclade could be the result of many unknown species yet
to be discovered, and also explain the low branch support
of this subclade. More importantly, the novel species
occupies a basal position with respect to C. spandovensis
and C. sergipiensis, indicating that the phylogenetic species
concept applies in the present case.
Since the two isolates of W. allomyrinae sp. nov. were
located in the Wickerhamiella clade, which contains six
sexual species assigned to the genus Wickerhamiella, special
efforts were made to induce their sexual state. However, the
strains did not produce ascospores or exhibit conjugation
on the most common sporulation media (YM agar, 5 %
malt extract agar, corn meal agar and YCBAS agar), alone
or mixed in pairs, at 15 uC or 25 uC for 1–4 weeks. In spite
of this result, the novel species was assigned to the genus
Wickerhamiella in conformance with the provisions of the
Melbourne Code (Norvell, 2011).
W. allomyrinae sp. nov. exhibited a narrow range of carbon
and nitrogen assimilation, which is typical for species in
the Wickerhamiella clade. However, the species could be
distinguished from the closely related species, C. musiphila,
C. spandovensis and C. sergipensis, by growth on vitaminfree medium and at 37 uC, which was positive for the novel
species and negative for the three closely related species
(Table 1). Furthermore, W. allomyrinae sp. nov. could be
distinguished from C. musiphila in terms of ability to
assimilate salicin, succinate and citrate, but not D-xylose.
The novel species also differed from C. spandovensis by
positive assimilation of salicin, inability to ferment glucose
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3857
Y.-C. Ren and others
67 Candida azyma CBS 6826T (EF536346)
74 Candida azyma UWOPS 95-693.4 (EF601042)
97
70
56
Candida sp. UWOPS 95-805.2 (EF601043)
71 Candida sp. UWOPS 95-863.2 (EF601041)
Candida azymoides UFMG-287T (DQ985171)
Candida parazyma UWOPS 99-724.2T (EF601046)
Candida sp. EJ3M02 (EF653941)
Candida alocasiicola AS 2.3484T (EU284106)
81 Wickerhamiella dulcicola UFMG-TOL15T (JQ180255)
52
Candida sp. LCF-01 NN12L02 (HQ623501)
89
Candida sp. UWOPS 03-446.4 (EU443389)
Wickerhamiella cachassae UFMG-D5L7T (JQ180256)
0.02
Candida sp. UL100 (HQ641271)
Candida vanderwaltii CBS 5524T (EU443388)
Candida sp. GE1L05 (FJ527060)
Wickerhamiella goesii IMUFRJ 52102T (JN790617)
61
90 Wickerhamiella occidentalis UWO(PS)91-698.4T (AF046037)
72
Wickerhamiella lipophila UWO(PS)91-681.3T (AF046040)
93
Wickerhamiella sp. 9E1 (AM946760)
99
Wickerhamiella australiensis UWOPS 05-260.2T (EF536348)
96
Wickerhamiella cacticola NRRL Y-27362T (AF046035)
Candida jalapaonensis UFMG-T05-210T (EU580139)
Wickerhamiella kiyanii FB1-1DASPT (JX978398)
82
Wickerhamiella pagnoccae UFMG-F18C1T (HQ593535)
54
Candida drosophilae UWO(PS)91-716.3T (EU443387)
70
Candida sp. 9A2 (FM178292)
73
Candida sp. UWO(PS)00-102.1 (AF313351)
100
Candida sp. EVN1238 CM122/05 (FR853155)
99
100 Candida sp. CBS 2275 (AY536215)
Candida pararugosa NRRL Y-17089T (U62306)
72
70
61
80
90
61
Candida sp. BG99-8-18-1-3-1 (AY242245)
Candida hasegawae NBRC 102566T (AB306510)
Wickerhamiella fructicola H10YT (JX978400)
61
Candida kazuoi NBRC 102565T (AB306509)
Candida galacta NRRL Y-17645T (DQ438239)
57
85
97
Candida sp. BG02-7-21-004C-1-1 (AY520293)
Candida bombiphila CBS 9712T (AJ620185)
Wickerhamiella domercqiae NRRL Y-6692T (DQ438240)
99
Wickerhamiella cf. domercqiae UWO(PS)00-107.1 (AF313369)
Wickerhamiella cf. domercqiae UWO(PS)00-192.1 (AF313368)
84 Candida sp. 107 (EF141077)
91
73
98
73
97
86
95
3858
Candida sp. SJ-1 (EF653272)
Candida sorbophila NRRL Y-7921T (DQ438229)
Candida infanticola CBS 11938T (HQ695009)
Candida sp. SN-102 (EF621560)
Candida sp. UWO(PS)00-136.3 (AF313356)
Candida musiphila AS 2.3479T (EU284104)
Wickerhamiella allomyrinae NYNU 13920T (KJ152751)
Candida sergipensis UFMG-R188T (AF397405)
Candida spandovensis NRRL Y-17761T (DQ438228)
Candida sp. BG02-7-18-027A-1-2 (AY520352)
Candida sp. BG01-7-26-006A-1-1 (AY242275)
Wickerhamiella slavikovae IMUFRJ 52096T (FJ463264)
Candida versatilis NRRL Y-6652T (DQ438242)
Starmerella bombicola NRRL Y-17069T (HQ111052)
Dipodascus magnusii NRRL Y-17563T (JQ689070)
85
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Wickerhamiella allomyrinae f.a., sp. nov.
Fig. 1. Phylogenetic tree derived from neighbour-joining analysis based on sequences of the D1/D2 domains of the LSU rRNA
gene, showing the placement of Wickerhamiella allomyrinae sp. nov. and other relevant species. Dipodascus magnusii NRRL
Y-17563T and Starmerella bombicola NRRL Y-17069T were used as outgroups. Bootstrap values of above 50 % are given at
nodes based on 1000 replications. Bar, 2 % sequence difference.
and galactose, and the lack of assimilation of L-arabinose,
xylitol, galactitol and ethanol. The novel species was easily
separated from C. sergipensis based on the ability to
assimilate citrate and succinate, and the inability to
assimilate xylitol, galactitol or ethanol.
Most species in the Wickerhamiella clade are highly
specialized nutritionally and ecologically, and some of
them have a strong association with flowers and floricolous
insects (Lachance & Kurtzman, 2011; Lachance et al., 2011;
Barbosa et al., 2012). For example, Lachance et al. (1998)
described five species in the Wickerhamiella clade, including two asexual taxa, Candida drosophilae and Candida
lipophila, isolated from flowers of Ipomoea acuminata and
its associated insect Drosophila floricola. Recent samplings
of yeasts revealed that the sugar cane plant may be a new
habitat for some yeasts in this clade. W. slavikovae and W.
goesii were isolated from sugar cane plants (Hagler et al.,
2013), whereas W. dulcicola and W. cachassae were isolated
from sugar cane juice and must, respectively (Badotti et al.,
2013). Of the 53 yeast strains isolated in this study,
Candida maltosa, Candida boleticola and Trichosporon
moniliiforme were the most frequently isolated species
from the gut of rhinoceros beetles. By comparison, only
two isolates of W. allomyrinae sp. nov. were obtained from
the environment we tested, making it more difficult to
speculate about the habitat of this novel species. However,
two other strains, namely Candida sp. BG02-7-18-027A-12 and Candida sp. BG01-7-26-006A-1-1, in the same
subclade have been isolated from the gut of beetles,
suggesting that this group of species may occur in beetle
guts and similar substrates in the Baotianman Reserve.
Description of Wickerhamiella allomyrinae Hui,
Ren, Wang, Chen & Ke sp. nov.
Wickerhamiella allomyrinae (al.lo.my.ri9nae. N.L. fem. gen.
n. allomyrinae referring to the genus of the host beetle,
Allomyrina dichotoma).
In YM broth after 3 days at 25 uC, cells are ovoid and
variable in size (2–463–5 mm), and occur singly or in
pairs. Budding is multilateral (Fig. 2). A sediment is
formed after one month, but no pellicle is observed. On
YM agar after 3 days at room temperature, colonies are
white, convex, smooth and opalescent, with an entire edge.
In Dalmau plates after 2 weeks on corn meal agar,
pseudomycelia or true mycelia are not formed. No asci
or signs of conjugation are observed after growth on the
most common sporulation media. Fermentation of glucose
is negative. Glucose, galactose, L-sorbose, sucrose, maltose,
salicin, arbutin, raffinose, glycerol, ribitol, glucitol, mannitol, D-galacturonate, succinate and citrate are assimilated.
Table 1. Physiological characteristics differentiating Wickerhamiella allomyrinae sp. nov. from closely related species
Species: 1, W. allomyrinae sp. nov.; 2, C. musiphila (data from Wang
et al., 2008); 3, C. spandovensis (Lachance et al., 2011); 4, C. sergipensis
(Lachance et al., 2011). +, Positive; 2, negative; D, delayed; S, slow;
V, variable; ND, not determined.
Characteristic
Fermentation of:
Glucose
Galactose
Assimilation of:
D-Xylose
L-Arabinose
Salicin
Xylitol
Galactitol
Succinate
Citrate
Ethanol
Other tests
Growth in vitamin-free medium
Growth at 37 uC
http://ijs.sgmjournals.org
1
2
3
2
2
2
2
+
V
S
2
D
V
2
2
2
+
2
+
+
+
2
2
+
2
2
+
+
2
+
+
ND
2
2
2
2
2
2
S
+
2
2
4
V
+
+
+
2
2
+
ND
2
Fig. 2. Photomicrographs of cells of Wickerhamiella allomyrinae
sp. nov. NYNU 13920T. Budding cells grown on YM agar for
3 days at 25 6C. Bar, 10 mm.
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Y.-C. Ren and others
No growth occurs with D-glucosamine, D-ribose, D-xylose,
L-arabinose, D-arabinose, L-rhamnose, a,a-trehalose, methyl
a-D-glucoside, cellobiose, melibiose, lactose, melezitose,
inulin, soluble starch, erythritol, xylitol, L-arabinitol,
galactitol, myo-inositol, 2-keto-D-gluconate, 5-keto-D-gluconate, D-gluconate, D-glucuronate, DL-lactate, methanol or
ethanol as sole carbon sources. Assimilation of nitrogen
compounds: positive for ethylamine, L-lysine and Dtryptophan, and negative for nitrate, nitrite, cadaverine,
creatine, creatinine, glucosamine and imidazole. Growth in
vitamin-free medium is positive. Growth is observed at
37 uC, but not at 40 uC. Growth in the presence of 10 %
NaCl plus 5 % glucose, 0.01 % cycloheximide and 1 % acetic
acid is negative. Acid formation on chalk agar is positive.
Starch-like compounds are not produced. Urease activity
and Diazonium Blue B reactions are negative.
T
The type strain, NYNU 13920 , was isolated from the gut
of A. dichotoma collected from the Baotianman National
Nature Reserve, Nanyan, Henan Province, China. It has
been deposited in the collection of the Yeast Division of
the Centraalbureau voor Schimmelcultures, Utrecht, the
Netherlands, as strain CBS 13167T and in the China Centre
of Industrial Culture Collection, Beijing, China, as strain
CICC 33031T. The Mycobank number is MB 809413.
two yeast species isolated from natural substrates. Int J Syst Evol
Microbiol 63, 3099–3103.
Hui, F. L., Niu, Q. H., Ke, T. & Liu, Z. (2012). Candida ficus sp. nov., a
novel yeast species from the gut of Apriona germari larvae. Int J Syst
Evol Microbiol 62, 2805–2809.
Hui, F. L., Chen, L., Chu, X. Y., Niu, Q. H. & Ke, T. (2013a).
Wickerhamomyces mori sp. nov., an anamorphic yeast species found
in the guts of wood-boring insect larvae. Int J Syst Evol Microbiol 63,
1174–1178.
Hui, F. L., Chen, L., Li, Z. H., Niu, Q. H. & Ke, T. (2013b). Metschnikowia
henanensis sp. nov., a new anamorphic yeast species isolated from
rotten wood in China. Antonie van Leeuwenhoek 103, 899–904.
Kimura, M. (1980). A simple method for estimating evolutionary rates
of base substitutions through comparative studies of nucleotide
sequences. J Mol Evol 16, 111–120.
Kurtzman, C. P. & Robnett, C. J. (1998). Identification and phylogeny
of ascomycetous yeasts from analysis of nuclear large subunit (26S)
ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73, 331–371.
Kurtzman, C. P. & Robnett, C. J. (2013). Relationships among genera
of the Saccharomycotina (Ascomycota) from multigene phylogenetic
analysis of type species. FEMS Yeast Res 13, 23–33.
Kurtzman, C. P., Fell, J. W., Boekhout, T. & Robert, V. (2011). Methods
for isolation, phenotypic characterization and maintenance of yeasts. In
The Yeasts, A Taxonomic Study, 5th edn, vol. 1, pp. 87–110. Edited by
C. P. Kurtzman, J. W. Fell & T. Boekhout. Amsterdam: Elsevier.
Lachance, M. A. & Kurtzman, C. P. (2011). Wickerhamiella van der
Walt (1973). In The Yeasts: A Taxonomic Study, 5th edn, vol. 2, pp.
891–897. Edited by C. P. Kurtzman, J. W. Fell & T. Boekhout.
Amsterdam: Elsevier.
Acknowledgements
This work was supported by the National Natural Science Foundation
of China (31370073) and the Research Planning Project of Basic and
Advanced Technology of Henan Province, China (122300410032).
References
Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z.,
Miller, W. & Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new
Lachance, M.-A., Rosa, C. A., Starmer, W. T., Schlag-Edler, B.,
Barker, J. S. F. & Bowles, J. M. (1998). Wickerhamiella australiensis,
Wickerhamiella cacticola, Wickerhamiella occidentalis, Candida drosophilae and Candida lipophila, five new related yeast species from
flowers and associated insects. Int J Syst Bacteriol 48, 1431–1443.
Lachance, M. A., Boekhout, T., Scorzetti, G., Fell, J. W. & Kurtzman,
C. P. (2011). Candida Berkhout (1923). In The Yeasts, a Taxonomic
Study, 5th edn, pp. 987–1278. Edited by C. P. Kurtzman, J. W. Fell &
T. Boekhout. Amsterdam: Elsevier.
generation of protein database search programs. Nucleic Acids Res 25,
3389–3402.
Nguyen, N. H., Suh, S. O., Erbil, C. K. & Blackwell, M. (2006).
Badotti, F., Silva, P. A. B., Mendonça, M. C., Gomes, F. C., Morais,
P. B., Lachance, M. A. & Rosa, C. A. (2013). Wickerhamiella dulcicola
Metschnikowia noctiluminum sp. nov., Metschnikowia corniflorae sp.
nov., and Candida chrysomelidarum sp. nov., isolated from green
lacewings and beetles. Mycol Res 110, 346–356.
sp. nov. and Wickerhamiella cachassae sp. nov., yeasts isolated from
cachaça fermentation in Brazil. Int J Syst Evol Microbiol 63, 1169–
1173.
Norvell, L. L. (2011). Fungal nomenclature. 1. Melbourne approves a
new code. Mycotaxon 116, 481–490.
Barbosa, A. C., Morais, C. G., Morais, P. B., Rosa, L. H., Pimenta,
R. S., Lachance, M. A. & Rosa, C. A. (2012). Wickerhamiella pagnoccae
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar,
S. (2011). MEGA5: molecular evolutionary genetics analysis using
sp. nov. and Candida tocantinsensis sp. nov., two ascomycetous yeasts
from flower bracts of Heliconia psittacorum (Heliconiaceae). Int J Syst
Evol Microbiol 62, 459–464.
maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28, 2731–2739.
Chen, L., Zhang, L., Li, Z. H. & Hui, F. L. (2013). Sympodiomycopsis
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. &
Higgins, D. G. (1997). The CLUSTAL_X windows interface: flexible
yantaiensis sp. nov., a basidiomycetous yeast isolated from insect frass.
Int J Syst Evol Microbiol 63, 3501–3505.
strategies for multiple sequence alignment aided by quality analysis
tools. Nucleic Acids Res 25, 4876–4882.
Dayo-Owoyemi, I., Rosa, C. A., Rodrigues, A. & Pagnocca, F. C.
(2014). Wickerhamiella kiyanii f.a., sp. nov. and Wickerhamiella
Urbina, H., Frank, R. & Blackwell, M. (2013). Scheffersomyces
fructicola f.a., sp. nov., two yeasts isolated from native plants of
Atlantic rainforest in Brazil. Int J Syst Evol Microbiol 64, 2152–2158.
cryptocercus: a new xylose-fermenting yeast associated with the gut
of wood roaches and new combinations in the Sugiyamaella yeast
clade. Mycologia 105, 650–660.
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach
van der Walt, J. P. & Liebenberg, N. V. (1973). The yeast genus
using the bootstrap. Evolution 39, 783–791.
Wickerhamiella gen. nov. (Ascomycetes). Antonie van Leeuwenhoek
39, 121–128.
Hagler, A. N., Ribeiro, J. R., Pinotti, T., Brandão, L. R., Pimenta, R. S.,
Lins, U., Lee, C. F., Hsieh, C. W., Lachance, M. A. & Rosa, C. A. (2013).
van der Walt, J. & van Kerken, A. (1960). Torulopsis domercqii nov.
Wickerhamiella slavikovae sp. nov. and Wickerhamiella goesii sp. nov.,
spec. Antonie van Leeuwenhoek 26, 314–316.
3860
Downloaded from www.microbiologyresearch.org by
International Journal of Systematic and Evolutionary Microbiology 64
IP: 88.99.165.207
On: Sun, 18 Jun 2017 10:58:17
Wickerhamiella allomyrinae f.a., sp. nov.
Wang, S. A., Jia, J. H. & Bai, F. Y. (2008). Candida alocasiicola sp. nov.,
Candida hainanensis sp. nov., Candida heveicola sp. nov. and Candida
musiphila sp. nov., novel anamorphic, ascomycetous yeast species
isolated from plants. Antonie van Leeuwenhoek 94, 257–265.
White, T. J., Bruns, T., Lee, S. & Taylor, J. W. (1990). Amplification
and direct sequencing of fungal ribosomal RNA genes for phylogenetics.
http://ijs.sgmjournals.org
In PCR Protocols: a Guide to Methods and Applications, pp. 315–322.
Edited by M. A. Innis, D. H. Gelfan, J. J. Sninsky & T. J. White. New
York: Academic Press.
Yarrow, D. (1998). Methods for the isolation, maintenance and
identification of yeasts. In The Yeasts, A Taxonomic Study, pp. 77–100.
Edited by C. P. Kurtzman & J. W. Fell. Amsterdam: Elsevier.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Sun, 18 Jun 2017 10:58:17
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