FEMS Yeast Research 2 (2002) 9^16
www.fems-microbiology.org
Sporobolomyces odoratus sp. nov., a new species in the
Sporidiobolus ruineniae clade
Elisabete Vale¤rio, Ma¤rio Gadanho, Jose¤ Paulo Sampaio *
Centro de Recursos Microbiolo¤gicos, Secc a‹o Auto¤noma de Biotecnologia, Faculdade de Cie“ncias e Tecnologia, Universidade Nova de Lisboa,
2829-516 Caparica, Portugal
Received 6 June 2001; received in revised form 4 October 2001; accepted 8 November 2001
First published online 7 December 2001
Abstract
This report presents the description of a new Sporobolomyces species, Sp. odoratus sp. nov. The new species was characterized by DNA
fingerprinting using the micro/minisatellite-primed PCR approach (MSP-PCR). A phylogenetic analysis using the D1/D2 region of the 26S
rDNA revealed that Sp. odoratus belongs to a clade that includes Rhodosporidium fluviale and Sporidiobolus ruineniae. An integrated
comparison with related species is presented. A number of presumptive strains of S. ruineniae were also investigated using the MSP-PCR
method and 26S rDNA sequence analysis. Mating experiments revealed, for the first time, that S. ruineniae includes heterothallic strains,
besides those already known to be self-fertile. : 2002 Federation of European Microbiological Societies. Published by Elsevier Science
B.V. All rights reserved.
Keywords : Basidiomycetous yeast; Taxonomy ; Sporobolomyces odoratus; Sporidiobolus ruineniae
1. Introduction
The anamorphic basidiomycetous genus Sporobolomyces
Kluyver and van Niel accommodates species that produce
bilaterally symmetrical ballistoconidia and have CoQ 10
or CoQ 10(H2) as their major ubiquinones. The typical
attributes of Sporobolomyces include incapacity to utilize
inositol and to produce amyloid compounds. In addition,
xylose is not present in whole-cell hydrolysates. The asexual genus Rhodotorula Harrison shares with Sporobolomyces all the characteristics mentioned above, except the capacity to produce ballistoconidia.
The polyphyletic nature of Sporobolomyces was revealed
by sequence analysis of 18S rDNA [1^3] and of the D1/D2
region of the 26S rDNA [4,5]. Within the Urediniomycetes, species like Sporobolomyces roseus Kluyver p van
Niel are related to the teleomorphic genus Sporidiobolus
Nyland, whereas species like Sp. oryzicola Nakase p Suzuki and S. elongatus Shivas and Rodrigues de Miranda
are a⁄liated to the Naohidea^Sakaguchia clade, and species such as Sp. xanthus (Nakase, Okada p Sugiyama)
* Corresponding author. Tel. : +351 (21) 2948300;
Fax: +351 (21) 2948530. E-mail address: jss@mail.fct.unl.pt (J.P. Sampaio).
Boekhout and Sp. lactophilus Nakase, Itoh, Suzuki p
Bandoni belong to the Agaricostilbum^Kondoa clade.
The type species of Sporobolomyces is Sporidiobolus salmonicolor (Fischer p Brebeck) Kluyver p van Niel and represents the anamorphic stage of S. salmonicolor Fell p
Tallman. It is possible that, in the future, Sporobolomyces
will be restricted to taxa related to the type species, in
order to decrease its present heterogeneity. However, any
taxonomic change has to take into account the proximity
between Sporidiobolus (and related Sporobolomyces species) and Rhodosporidium (and related Rhodotorula species). In fact, sequence analysis of 26S rDNA has indicated
a close relationship between Sporidiobolus ruineniae Holzschu, Tredick p Pha¡, Sporidiobolus microsporus Higham
ex Fell, Blatt p Statzell-Tallman, Rhodosporidium azoricum Sampaio p Gadanho, Rhodosporidium £uviale Fell,
Kurtzman, Tallman p Buck and R. lusitaniae Fonseca
and Sampaio [6,7]. These ¢ve species belong to the £uviale^ruineniae group, one of the three clades in which
the species of the genera Sporidiobolus and Rhodosporidium, together with their close asexual relatives, are divided
[7]. Recently, Takashima and Nakase [8] described Sporobolomyces nylandii Takashima p Nakase and Sporobolomyces poonsookiae Takashima p Nakase, two anamorphic
species that belong to the £uviale^ruineniae clade. This
report presents the description of an additional Sporobo-
1567-1356 / 02 / $22.00 : 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
PII : S 1 5 6 7 - 1 3 5 6 ( 0 1 ) 0 0 0 5 4 - X
10
Table 1
List of strains used in MSP-PCR ¢ngerprinting
Strain
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
IGC
Sp. odoratus
IGC 5696
PDA contamination, Monte de Caparica, Portugal
Sp. odoratus
Sp. odoratus
Sp. poonsookiae Takashima p Nakase
IGC 5697
IGC 5698
IGC 5699T
Leaf of PlatanusUacerifolia, Monte de Caparica, Portugal
Leaf of lemon tree, Algueira‹o, Portugal
Dead leaf of Mangifera indica, Thailand
5062T
5084
4701T
4641T
4599
4642
5617T
4201T
4211
5676
5677
5692
5660T
5596
5693T
5694T
5695
Strain origin
CBS 6568T
CBS
CBS
CBS
CBS
CBS
7041T
5001T
5000
5004
5078
CBS 5811T
CBS 5624
JCM 10213T
ZP 351
JCM 10207T
Isolation source
Observations
Soil, Azores, Portugal
Soil, Azores, Portugal
Brackish water, USA
Woodland soil, Lisbon, Portugal
Dry leaf, Arra¤bida, Portugal
Plant litter, Arra¤bida, Portugal
Herbaceous culm, Jamaica
Leaf of Malpighia coccigera, Indonesia
Leaf of M. coccigera, Indonesia
Unknown substrate, Indonesiaa
Unknown substrate, Indonesiaa
Garden soil, Taiwan
Dung of goat, Pakistan
Air, Japanb
Dead leaf of Oryza sativa, Thailand
Basidiocarp of Myxarium nucleatum, Sesimbra, Portugal
Leaf of Acer monspessulanum, Arra¤bida, Portugal
Mating type
Mating type
Self-fertile
Self-fertile
Self-fertile
Self-fertile
Self-fertile
Self-fertile
Self-fertile
Mating type
Anamorphic
Mating type
A1
A2
A1
A2
Absence of ballistoconidia
Isolated by J.P. Sampaio, December 1996
Isolated by A. Fonseca and J. Ina¤cio,
November 1998
Isolated by J.P. Sampaio and E. Vale¤rio,
November 1999
Isolated by A. Fonseca, May 2000
Isolated by A. Fonseca, October 1986
IGC, Portuguese Yeast Culture Collection, FCT-UNL, Portugal; CBS, Centraalbureau voor Schimmelcultures, Yeast Division, Utrecht, The Netherlands ; JCM, Japan Collection of Microorganisms, RIKEN, Japan; ZP, personal collection of J.P. Sampaio.
a
Isolated by J. Ruinen and deposited at CBS as S. roseus.
b
Type strain of Pichia rosea; maintained at CBS as Rhodotorula graminis.
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
Species
Rhodosporidium azoricum Sampaio p Gadanho
R. azoricum
R. £uviale Fell, Kurtzman, Tallman p Buck
R. lusitaniae Fonseca p Sampaio
R. lusitaniae
R. lusitaniae
S. microsporus Higham ex Fell, Blatt p Statzell-Tallman
S. ruineniae Holzschu, Tredick p Pha¡ var. ruineniae
S. ruineniae var. ruineniae
S. ruineniae var. ruineniae
S. ruineniae var. ruineniae
S. ruineniae var. ruineniae
S. ruineniae var. coprophilus Kurtzman p Fell
S. ruineniae var. coprophilus
Sp. nylandii Takashima p Nakase
Sp. odoratus Sampaio, Fonseca p Vale¤rio
Sp. odoratus
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
lomyces species in this clade, Sp. odoratus sp. nov. A comparison with closely related taxa is presented.
2. Materials and methods
2.1. Isolations, morphological characterization and studies
of sexual compatibility
For the isolations, the ballistoconidium-fall method was
employed. For microscopy, cultures were grown on MYP
agar (malt extract 0.7% w/v, yeast extract 0.05% w/v, soytone 0.25% w/v and agar 1.5% w/v) at room temperature
(20^23‡C) and studied with an Olympus BX50 microscope, using phase contrast optics. For determination of
sexual compatibility, pairs of 2^4-day-old cultures were
crossed on SG agar (soytone 0.2% w/v, glucose 0.2% w/v
and agar 1.5% w/v), incubated at room temperature and
examined for production of mycelium and teliospores after
1 week.
2.2. Physiological characterization
Physiological and biochemical characterization was performed according to the techniques described by Yarrow
[9]. Additional assimilation tests were performed using aldaric acids and aromatic compounds, as described by Fonseca [10] and Sampaio [11], respectively.
11
3. Results and discussion
Between 1986 and 2000, ¢ve strains of an anamorphic
pink-colored ballistoconidial yeast were isolated from several localities in Portugal (Table 1). All the isolates were
light pink, dull, exhaled a peculiar odor and produced
large quantities of forcibly discharged propagules. The
latter characteristic resulted in the formation of numerous
small colonies that covered the entire surface of the culture
medium. In addition, ballistoconidia were frequently produced at the end of long stalks. All strains were unable to
utilize melezitose and to grow at 35‡C. This combination
of physiological traits occurs in some species of the £uviale^ruineniae clade, such as S. microsporus, S. ruineniae
and R. lusitaniae. Therefore, the isolates were initially
compared with the species of this group, regardless of their
ability to produce ballistoconidia.
The indication that the ¢ve strains were conspeci¢c and
distinct from the remaining species of the £uviale^ruineniae
clade was obtained in DNA ¢ngerprinting experiments
using MSP-PCR. This investigation was conducted using
the primers M13 and (GTG)5 and the banding patterns
obtained are depicted in Fig. 1. It can be observed that the
pro¢les of the various strains of Sp. odoratus are similar,
but clearly di¡erent from those of the remaining species in
the £uviale^ruineniae clade. In order to determine the phy-
2.3. PCR ¢ngerprinting and rDNA sequence analysis
For PCR ¢ngerprinting, the DNA extraction protocol,
primers, PCR and electrophoresis conditions and gel image analysis procedures were those reported by Sampaio et
al. [12]. A total of 21 strains representing the various species of the £uviale^ruineniae clade were investigated using
the micro/minisatellite-primed PCR approach (MSPPCR). The list of cultures used in this study, and information related to them, is presented in Table 1. For rDNA
sequence analysis, total DNA was extracted using the protocol of Sampaio et al. [12] and ampli¢ed using primers
ITS5 and LR6. Cycle sequencing of the 600^650-bp region
at the 5P end of the 26S rDNA D1/D2 domain employed
forward primer 5P-GCA TAT CAA TAA GCG GAG
GAA AAG and reverse primer 5P-TCC TCC GCT TAT
TGA TAT GC. Sequences were obtained with an Amersham Pharmacia ALF Express II automated sequencer
using standard protocols. Alignments were made with
MegAlign (Dnastar Inc., Madison, WI, USA) and visually
corrected. Phylogenetic trees were computed with PAUP
version 4.0 using the neighbor-joining method [13]. Distances between sequences were calculated using Kimura’s
two-parameter model [14]. Bootstrap analysis [15] was
based on 1000 random resamplings.
Fig. 1. MSP-PCR ¢ngerprints of Sp. odoratus and remaining species of
the £uviale^ruineniae clade generated with primers M13 (a) and (GTG)5
(b). M, molecular size marker (V DNA cleaved with HindIII and
xX174 DNA cleaved with HaeIII).
12
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
logenetic position of the new species, the nucleotide sequence of the D1/D2 region of the 26S rDNA was determined (Fig. 2). Sequence analysis con¢rmed that Sp. odoratus represents a distinct species and that it belongs to the
£uviale^ruineniae clade.
Currently, the £uviale^ruineniae clade comprises three
species of Sporobolomyces and presents two particularities.
It includes species that are able to produce ballistoconidia
together with species that do not present this trait. This
issue has been addressed by Fell et al. [6] and, more recently, by Gadanho et al. [7]. Secondly, the basidia of
some sexual species (R. lusitaniae, S. ruineniae and S. microsporus) are formed at the end of a stalk [7], which does
not occur in any of the species belonging to the other two
Fig. 2. Phylogenetic tree of S. odoratus and related taxa of the genera Sporobolomyces, Rhodotorula, Sporidiobolus and Rhodosporidium, obtained by
neighbor-joining analysis of the D1/D2 region of the 26S rDNA. Rhodotorula minuta and Saccharomyces cerevisiae were included to root the tree. Percentage bootstrap values of 1000 replicates are given at each node. GenBank accession numbers are indicated after species designation (names in boldface correspond to sequences determined in this study; T, type strain).
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
13
Table 2
Physiological characteristics of the strains of Sp. odoratus
Carbon compounds
IGC 5694T
IGC 5695
IGC 5696
IGC 5697
IGC 5698
D-Glucose
+
3
+
3
3
+
D
D
3
+
3
+
3
3
+
+
3
3
+
3
3
3
+
3
+
D
+
+
3
3
+
DW
3
3
+
3
3
3
3
3
3
3
3
+
+
3
3
+
3
+
3
3
3
3
3
+
+
+
3
3
+
D
D
3
+
3
+
3
3
+
+
3
3
+
3
3
3
D
3
+
D
+
+
3
3
+
+
3
3
+
3
+
3
3
3
3
3
3
+
+
3
3
+
3
+
3
3
3
3
3
+
+
+
3
3
+
3
3
3
+
3
+
3
3
3
+
3
3
+
3
3
3
D
3
+
3
+
+
3
3
DW
D
3
3
+
3
D
3
3
3
3
3
3
+
D
3
3
+
3
+
3
3
3
3
3
+
+
+
3
3
+
D
3
3
+
3
+
3
3
+
+
3
3
+
3
3
3
D
3
+
D
+
+
3
3
+
+
3
3
+
3
+
3
3
3
3
3
3
+
+
3
3
+
3
+
3
3
3
3
3
+
3
+
3
3
+
D
3
3
+
3
+
3
3
3
+
3
3
+
3
3
3
+
3
+
3
+
W
3
3
+
W
3
3
DW
3
3
3
3
3
3
3
3
+
3
3
3
+
3
D
3
3
3
3
3
+
+
3
3
3
+
+
3
3
3
+
+
3
3
3
+
+
3
3
3
+
+
3
3
3
D-Galactose
L-Sorbose
D-Glucosamine
D-Ribose
D-Xylose
L-Arabinose
D-Arabinose
L-Rhamnose
Sucrose
Maltose
K,K-Trehalose
Methyl-K-D-glucoside
Cellobiose
Salicin
Arbutin
Melibiose
Lactose
Ra⁄nose
Melezitose
Inulin
Soluble starch
Glycerol
Erythritol
Ribitol
Xylitol
D-Glucitol
D-Mannitol
Galactitol
Inositol
Glucono-N-lactone
D-Gluconic acid
D-Glucuronic acid
D,L-Lactic acid
Succinic acid
Citric acid
L-Malic acid
L-Tartaric acid
D-Tartaric acid
m-Tartaric acid
Saccharic acid
Mucic acid
Methanol
Ethanol
Vanillic acid
Veratric acid
Ferulic acid
p-Hydroxybenzoic acid
m-Hydroxybenzoic acid
Protocatechuic acid
Catechol
Gallic acid
Salicylic acid
Gentisic acid
Phenol
Nitrogen compounds
Nitrate
Nitrite
Ethylamine
L-Lysine
Cadaverin
D, delayed; W, weak results.
14
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
Table 2 (continued).
Carbon compounds
IGC 5694T
IGC 5695
IGC 5696
IGC 5697
IGC 5698
Creatine
Creatinine
Other tests
Growth in vitamin-free medium
Growth with 0.01% cycloheximide
Growth with 0.1% cycloheximide
Growth at 30‡C
Growth at 35‡C
Formation of starch-like compounds
Splitting of arbutin
Hydrolysis of urea
Colour reaction with diazonium blue B
3
3
3
3
3
3
3
3
3
3
+
3
3
3
3
3
+
+
+
+
3
3
3
3
3
+
+
+
+
3
3
+
3
3
+
+
+
+
3
3
3
3
3
+
+
+
+
3
3
3
3
3
+
+
+
D, delayed; W, weak results.
are discrepant between Sp. odoratus and R. lusitaniae. In
contrast to R. lusitaniae, the new species utilizes sucrose,
ra⁄nose and does not grow with galactitol, ferulic acid,
p-hydroxybenzoic acid, gallic acid and catechol.
Since the MSP-PCR approach is well suited to characterize large sets of isolates, we also investigated a group of
strains suspected to belong to S. ruineniae (IGC 5596, IGC
5676, IGC 5677 and IGC 5692). Three strains, IGC 5676,
IGC 5677 and IGC 5692, showed DNA banding pro¢les
similar to IGC 4201T and IGC 4211, the two con¢rmed
representatives of S. ruineniae var. ruineniae, whereas the
patterns of IGC 5596 were comparable to those of IGC
5660T , the single known strain of S. ruineniae var. coprophilus Kurtzman p Fell (Fig. 1). Among the variety rui-
clades (toruloides^glutinis and johnsonii^pararoseus). The
phylogenetic tree in Fig. 2 indicates that the closest relative of Sp. odoratus is Sp. nylandii. The two species have
11 di¡erent bases in the D1/D2 region of the 26S rDNA.
S. poonsookiae, the third anamorphic species in this clade,
has 25 di¡erent bases towards Sp. odoratus.
The physiological and biochemical properties of Sp. odoratus are depicted in Table 2 and a comparison with other
ballistoconidia-producing taxa is presented in Table 3. The
closest relatives of Sp. odoratus are Sp. nylandii, Sp. poonsookiae and R. lusitaniae (Fig. 2). The new species does not
utilize D-ribose, cellobiose and catechol and is not able to
grow in the presence of 0.1% cycloheximide, in contrast to
Sp. nylandii and Sp. poonsookiae. Several physiological tests
Table 3
Salient physiological di¡erences between Sp. odoratus and related ballistoconidia-producing taxa
Cluster 1 toruloides^glutinis
Sp. alborubescens Derx (CBS 482T )
Cluster 2 £uviale^ruineniae
S. ruineniae Holzschu, Tredick p Pha¡ (CBS 5001T ,
CBS 5000)
S. microsporus Higham ex Fell, Blatt p Statzell-Tallman
(CBS 7041T )
Sp. nylandii Takashima p Nakase (IGC 5693T )
Sp. odoratus Sampaio, Fonseca p Vale¤rio (IGC 5694T ,
IGC 5695, IGC 5696, IGC 5697, IGC 5698)
Sp. poonsookiae Takashima p Nakase (IGC 5699T )
Cluster 3 johnsonii^pararoseus
Sp. roseus Kluyver p van Niel (JCM 5353LT )
Sp. marcillae Santa Mar|¤a (CBS 4217T )
Sp. ruberrimus Yamazaki p Fujii (CBS 7500AUT ,
CBS 7501)
S. pararoseus Fell p Tallman (CBS 491T , CBS 484,
CBS 4216, IGC 4622)
S. salmonicolor Fell p Tallman (CBS 490T , CBS 2630,
CBS 2643, IGC 4351, IGC 4558, IGC 4623)
S. johnsonii Nyland (CBS 5470T , IGC 4353)
Sp. holsaticus Windish ex Yarrow p Fell (CBS 1522T )
Malt.
Ribo.
Melz.
Cellob.
Starch
30‡C
35‡C
0.1% Cx.
AmH. Cat.
+
+
+
3
3
+
+
+
+
3
+
+
3
+
3
+
3
+
+
+
3
+
3
+
3
+
3
+
+
+
+
3
+
3
+
3
+
3
3*
3
+
V
+
3
+
3
3
3
+
3
3
+
3
+
3*
+
3
+
+
+
+
+
+
+
+
3
+
+
+
+
+
V
+
+
+
3
3
3
3
3
3
3
3
3
3
+
3
3
3
3
+
3
+
+
+
+
3
3
+
3
V
V
V
V
V
+
+
+
V
+
+
+
V
+
V
+
+
+
+
3
+
+
+
+
+
+
3
3
+
+
Results: (+) positive (including delayed and weak results), (3) negative and (V) variable.
Data are from the present study and from the Portuguese Yeast Culture Collection database and correspond to the strains indicated between brackets ;
* positive results in the original description [8]. Abbreviations: Malt., Maltose ; Ribo., D-Ribose; Melz., Melezitose ; Cellob., Cellobiose; 0.1% Cx.,
growth in the presence of 0.1% cycloheximide; AmH., m-hydroxybenzoic acid; Cat., catechol.
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
neniae, two strains (IGC 5676 and IGC 5677), previously
identi¢ed as Sp. roseus [16], produced numerous ballistoconidia and strain IGC 5692 produced ballistoconidia very
rarely. With respect to the variety coprophilus, IGC 5596
did not form ballistoconidia similarly to IGC 5660T , the
type strain of this variety. The strain IGC 5596 was originally described as the type of Pichia rosea Nishiwaki and
was received from the CBS (Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands) culture collection
as Rhodotorula graminis di Menna. Interestingly, all the
seven isolates of S. ruineniae were collected in Asia (Indonesia, Taiwan, Japan and Pakistan) and, although we have
recently studied large sets of ballistoconidial yeasts from
Portugal using the MSP-PCR approach, none of them
belonged to S. ruineniae. Since these four new members
of S. ruineniae were not self-fertile, as is the case for the
previously characterized strains of this species [17], we investigated their mating compatibility in crossing experiments. From the six pairings inoculated on SG agar, mycelium and teliospores were observed in the crossing of
IGC 5676 and IGC 5692. Besides incomplete clamp connections, true clamps were also observed. The teliospores
originated from transversally septate basidia similar to
those already observed in the homothallic strains. These
results indicate that S. ruineniae includes both homothallic
and heterothallic strains, a situation that was not reported
previously for this species. Description of a sexual yeast
species based solely on homothallic isolates followed, later,
by the unravelling of heterothallic strains based on molecular typing methods occurred previously for Rhodosporidium kratochvilovae Hamamoto, Sugiyama p Komagata
[12]. The examples of S. ruineniae and R. kratochvilovae
show the importance of molecular typing in the discovery
of new anamorph^teleomorph connections. Moreover, our
¢ndings also demonstrate the relevance of population-focussed studies rather than type strain-oriented investigations. We designate IGC 5676 as the mating type A1 and
IGC 5692 as the mating type A2 of S. ruineniae var. ruineniae. The other two strains (IGC 5677 and IGC 5596) did
not exhibit any sign of sexual behavior and are presently
considered as anamorphic strains of this species. To con¢rm our identi¢cations, we determined the D1/D2 sequences of the two mating strains and also that of IGC 5596.
No di¡erences were detected in comparison with the published sequences of S. ruineniae (Fig. 2). With respect to
conventional phenotypic criteria, the L-rhamnose test and
the mode of teliospore germination can be used to distinguish the two varieties of S. ruineniae [18]. We were able to
con¢rm growth with this carbon compound for the strains
of the variety ruineniae (weak results for IGC 5677, delayed for IGC 5692 and vigorous for IGC 5676, 4201 and
4211) and absence of growth for the two strains of the
variety coprophilus. The reported heterogeneity in teliospore germination could not be investigated because
none of the strains of the variety coprophilus produced
teliospores.
15
3.1. Latin diagnosis of Sporobolomyces odoratus Sampaio,
Fonseca p Vale¤rio sp. nov.
Post 2 dies in MYP agar cellulae ovoideae ad cylindraceae (2) 3^5U5^8 (9) Wm. Ballistoconidia ovoidea ad reniformia, 3^4U6^10 Wm, ex sterigmatibus longis (2^
3U10^70 Wm) abundanter oriunda. Mycelium verum non
formatur. Post unum mensem ad 20^22‡C cultura in striis
subrosea, hebes, super¢cie laevi vel ex parte rugosa, textura butyracea. Assimilatio maltosi, D-ribosi, melezitosi,
cellobiosi deest, nullum incrementum ad 35‡C. Cultura
odore peculiari.
3.2. Description of Sporobolomyces odoratus Sampaio,
Fonseca p Vale¤rio sp. nov.
Yeast cells after 2 days on MYP agar ovoid to cylindrical measuring (2) 3^5U5^8 (9) Wm (Figs. 3 and 4). Ballistoconidia abundantly produced, ovoid to reniform, measuring 3^4U6^10 Wm and originating at the end of usually
long (2^3U10^70 Wm) sterigmata (Figs. 3 and 4). True
mycelium is not formed. After 1 month at 20^22‡C, the
streak culture is pale-pink, dull, the surface is smooth or
slightly wrinkled and the texture is butyrous. A salient
feature is the production of a peculiar odor. Unable to
ferment glucose. Additional physiological and biochemical
characteristics are depicted in Table 2.
Fig. 3. Line drawings of yeast cells, sterigmata and ballistoconidia of
Sp. odoratus grown on MYP agar at room temperature for 2^3 days.
a: Cell with long sterigma and immature ballistoconidium (IGC 5697) ;
b: budding yeast cells and formation of ballistoconidia (IGC 5694T );
c: released ballistoconidia of IGC 5694T and IGC 5697. Bar = 10 Wm.
16
E. Vale¤rio et al. / FEMS Yeast Research 2 (2002) 9^16
References
Fig. 4. Micrographs of ballistoconidia-forming cells and ballistoconidia
of Sp. odoratus grown on MYP agar at room temperature for 2^3 days.
a: IGC 5697; b: IGC 5694T . Bar = 10 Wm.
3.2.1. Etymology
The speci¢c epithet odoratus ( = fragrant) refers to the
peculiar odor of the cultures of this species.
3.2.2. Origin, type and deposits
IGC 5694 was isolated by J.P. Sampaio from a basidiocarp of Myxarium nucleatum Wallroth collected at Quinta
dos Pinheiros, Sesimbra, 30 km south of Lisbon, Portugal,
in December 1996. This strain was deposited lyophilized
(holotype) and living (ex-type) in the Portuguese Yeast
Culture Collection, Caparica, Portugal. Information on
the origin of other strains of this species is presented in
Table 1.
Acknowledgements
M.G. was supported by a Grant SFRH/BD/1170/2000.
The authors are indebted to Dr. Chee-Jen Chen (Southern
Taiwan University of Technology, Taiwan) for sending a
strain (IGC 5692) of S. ruineniae, to Dr. M. Hamamoto
(Japan Collection of Microorganisms, Japan) for sending
the type strains of Sp. nylandii and Sp. poonsookiae, to
Prof. Isabel Spencer-Martins for critical reading of the
manuscript and to Dr. Michael WeiM (University of Tu«bingen, Germany) for preparing the Latin diagnosis.
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