International Journal of Systematic and Evolutionary Microbiology (2013), 63, 1646–1652
DOI 10.1099/ijs.0.044669-0
Winogradskyella psychrotolerans sp. nov., a marine
bacterium of the family Flavobacteriaceae isolated
from Arctic sediment
Z. Begum, T. N. R. Srinivas, P. Manasa, B. Sailaja, B. Sunil, S. Prasad
and S. Shivaji
Correspondence
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500 007, India
S. Shivaji
[email protected]
A novel Gram-negative, rod–coccus shaped, non-motile, strain, RS-3T, was isolated from a
sediment sample collected from the marine transect of Kongsfjorden, Ny-Ålesund, Svalbard,
Arctic. Colonies and broth cultures were yellowish in colour due to the presence of carotenoids.
Strain RS-3T was positive for oxidase, aesculinase, caseinase, gelatinase and urease activities
and negative for amylase, catalase, lipase, lysine decarboxylase, ornithine decarboxylase, DNase
and b-galactosidase activities. The predominant fatty acids were iso-C15 : 0 (18.0), anteiso-C15 : 0
(16.8), iso-C15 : 1 G (14.2), anteiso-C15 : 1 A (6.0) and iso-C15 : 0 3-OH (6.8). Strain RS-3T
contained MK-6 (72.42 %) and MK-7 (27.58 %) as the major respiratory quinones and
phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids make up the
polar lipid composition. The DNA G+C content of strain RS-3T was 34.7±1.2 mol%. The 16S
rRNA gene sequence analysis indicated that Winogradskyella pacifica and Winogradskyella
thalassocola are the most closely related species with sequence similarities to the type strains of
these species of 98.5 and 97.7 %, respectively. However, DNA–DNA hybridization with
Winogradskyella pacifica KCTC 22997T and Winogradskyella thalassocola DSM 15363T
showed a relatedness of 22 and 42.5 % with respect to strain RS-3T. Based on the DNA–DNA
hybridization values, phenotypic and chemotaxonomic characteristics and phylogenetic inference,
strain RS-3T is proposed as a novel species of the genus Winogradskyella, for which the name
Winogradskyella psychrotolerans sp. nov. is proposed. The type strain of Winogradskyella
psychrotolerans sp. nov. is RS-3T (5CIP 110154T5NBRC 106169T). An emended description
of the genus Winogradskyella is provided.
The genus Winogradskyella is a part of the family Flavobacteriaceae, phylum ‘Bacteroidetes’. The cells of species of
the genus Winogradskyella are rod-shaped, Gram-negative,
motile by gliding, capable of forming network-like structures, non-endospore forming, produce non-diffusible
yellow or orange non-flexirubin type of pigment, strictly
aerobic or facultatively anaerobic, chemo-organotrophic,
positive for cytochrome oxidase (variable), catalase and
alkaline phosphatase activities and nitrate reduction and
can hydrolyse gelatin, starch and DNA. The main cellular
fatty acids are iso-C15 : 0, anteiso-C15 : 0, iso-C15 : 1, iso-C16 : 0
3-OH and iso-C17 : 0 3-OH. The polar lipids included
phosphatidylethanolamine, one or two unknown aminolipids and one unknown lipid. The major respiratory
Abbreviations: FAMEs, fatty acid methyl esters; NJ, neighbour joining;
PE, phosphatidylethanolamine.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene
sequence of strain RS-3T is FN377721.
A supplementary figure is available with the online version of this paper.
1646
quinone is menaquinone 6 (MK-6) and DNA G+C
content is 30.2–37 mol% (Ivanova et al., 2010;
Nedashkovskaya et al., 2005; Nedashkovskaya et al., 2012;
Yoon et al., 2011). Currently, the genus Winogradskyella
comprises 14 species with validly published names. The
species of the genus are widely distributed in different
marine habitats like seawater (Kim & Nedashkovskaya
2010; Yoon et al., 2011), marine algae (Nedashkovskaya
et al., 2005; Nedashkovskaya et al., 2012), starfish (Ivanova
et al., 2010), marine sediments (Romanenko et al., 2009),
sea urchin (Nedashkovskaya et al., 2009), sponge (Lau
et al., 2005) and protein-enriched surface seawater
(Pinhassi et al., 2009). In the present study, a strain RS3T was isolated from marine sediment collected from
Kongsfjorden, Ny-Ålesund, Svalbard, Arctic (Srinivas et al.,
2009). Phylogenetic analysis based on the 16S rRNA gene
sequence analysis indicated that strain RS-3T clustered
within the genus Winogradskyella. Based on phenotypic
and chemotaxonomic characteristics with the nearest
phylogenetic neighbours, it is proposed that strain RS-3T
represents a novel species.
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Winogradskyella psychrotolerans sp. nov.
Strain RS-3T was isolated from a sediment sample (brown,
clay-like, sticky with algae) collected from 4.2 m depth of
Kongsfjorden, Ny-Ålesund, Svalbard (78u 569 07.90 N 11u
529 54.90 E), in the Arctic on 9 August 2007. The sample
that yielded strain RS-3T had a pH of 7.6. The surface and
bottom water temperature at the RS sampling site at the
time of collection was 5.2 uC. For isolation of bacteria,
100 mg of the sediment sample was suspended in 900 ml
sterile normal saline and subjected to shaking for 2 h at
20 uC. The supernatant was serially diluted and 100 ml was
plated on Zobel marine agar (MA, HIMEDIA) and incubated at 10 uC for 15 days. Different morphotypes were
purified and maintained on MA plates.
Laboratories) and sequenced as described previously (Lane,
1991). The resultant almost complete sequence of the 16S
rRNA gene (1510 nt) was subjected to BLAST sequence
similarity search (Altschul et al., 1990) to identify the most
closely related taxa. All the 16S rRNA gene sequences
belonging to the genus Winogradskyella were downloaded
from the database (http://www.ncbi.nlm.nih.gov), aligned
using the CLUSTAL_X program (Thompson et al., 1997) and
the alignment corrected manually. Phylogenetic trees were
constructed using the maximum-likelihood (ML) and
neighbour joining (NJ) (Saitou & Nei, 1987) tree-making
algorithms, using the PhyML program (Guindon et al.,
2005) and the PHYLIP package, version 3.5 (Felsenstein,
1993), respectively. The resultant tree topologies were evaluated by bootstrap analysis based on 1000 resamplings
using the SEQBOOT and CONSENSE programs in the PHYLIP
package. Pair-wise evolutionary distances were computed
using the DNADIST program with the Kimura two-parameter
model (Kimura, 1980).
The culturable bacterial count (c.f.u.) obtained was
0.86104 g21 sediment. A total of 16 morphotypes were
obtained. Out of this one strain each belonged to the
genera Arthrobacter, Marinobacterium and Winogradskyella;
three strains to the genus Pseudoalteromonas; and five
strains each to the genera Pseudomonas and Shewanella
(Srinivas et al., 2009). The yellowish-coloured colony
which belonged to the genus Winogradskyella was selected
and characterized. For 16S rRNA gene sequencing, DNA
was prepared using a microbial DNA isolation kit (Mo Bio
0.01
Strain RS-3T shared the highest 16S rRNA gene pairwise
sequence similarity with Winogradskyella pacifica and
Winogradskyella thalassocola (98.5 % and 97.7 % respectively). Phylogenetic analyses further indicated that strain
Winogradskyella exilis 022-2-26T (FJ595484)
Winogradskyella poriferorum UST030701-295T (AY848823)
Winogradskyella echinorum KMM 6211T (EU727254)
Winogradskyella ulvae KMM 6390T (HQ456127)
Winogradskyella eximia KMM 3944T (AY521225)
Winogradskyella epiphytica KMM 3906T (AY521224)
Winogradskyella lutea A73T (FJ919968)
100
Winogradskyella arenosi R60T (AB438962)
Winogradskyella rapida SCB36T (U64013)
Winogradskyella thalassocola KMM 3907T (AY521223)
63
100
Winogradskyella pacifica KMM 6019T (GQ181061)
Winogradskyella psychrotolerans RS-3 T (FN377721)
Salegentibacter salarius ISL-6T (EF486353)
Fig. 1. Phylogenetic tree based on 16S rRNA gene sequences showing the relationship of Winogradskyella psychrotolerans
RS-3T with species of the genus Winogradskyella and other closely related members of the family Flavobacteriaceae. The
phylogenetic tree was constructed using the maximum-likelihood method. Numbers at nodes are bootstrap values.
Salegentibacter salarius ISL-6T (EF486353) was used as an outgroup. Bar, 0.01 substitutions per alignment position.
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Z. Begum and others
Table 1. Features that distinguish Winogradskyella psychrotolerans RS-3T from the closely related species of the genus
Winogradskyella
Strains: 1, Winogradskyella psychrotolerans RS-3T; 2, Winogradskyella pacifica KCTC 22997T; 3, Winogradskyella thalassocola DSM 15363T. Data for
all three taxa is from the present study. All strains were Gram-negative, yellow pigmented, positive for catalase and oxidase activities, casein and
gelatin hydrolysis and phenylalanine, glutamic acid and cysteine utilization. All strains were negative for lysine decarboxylase, ornithine
decarboxylase and phenylalanine deaminase activities, H2S and indole production, Voges–Proskauer and methyl red reactions, agar, DNA and
Tween-60 hydrolysis and failed to utilize xylose, adonitol, rhamnose, cellobiose, raffinose, trehalose, maltose, galactose, inulin, sodium gluconate,
salicin, dulcitol, inositol, methyl-a-D-glucoside, ribose, melezitose, methyl-a-D-mannoside, xylitol, D-arabinose and sorbose. All strains were
sensitive to norfloxacin (10), lomefloxacin (30), roxithromycin (30), chloramphenicol (30), erythromycin (15) and tetracycline (30) and resistant to
colistin (10) and nalidixic acid (30). +, Positive; 2, negative; W, weak. R, resistant; S, sensitive. PE, phosphatidylethanolamine; AL1–AL2,
unidentified aminolipids; L1–L3, unidentified lipids.
Characteristic
Cell shape
Cell size (mm)
Motility
Temperature growth range
(uC)
Optimum growth
temperature (uC)
NaCl range (%, w/v)
Optimum NaCl
concentration (%, w/v)
pH growth range
Optimum pH
Nitrate reduction
Hydrolysis of:
Aesculin
Starch
ONPG
Urea
Utilization of:
Citrate
Malonate
Arabinose
Sucrose
Glucose
Lactose
Fructose
Melibiose
L-Arabinose
Mannose
Glycerol
Glucosamine
Sorbitol
Mannitol
Aspartic acid
Alanine
Leucine
Isoleucine
Tryptophan
Methionine
Histidine
Valine
Antibiotic susceptibility
(mg/disc):
Co-trimoxazole (25)
Ampicillin (10)
1648
1
2
3
Rod–coccus
0.4–0.660.5–1.0
2
4–30
Rod
0.4–0.561–2.3
+
4–30
Rod
0.5–0.764–7.3
+
4–37
18
21-23
1–6
2
1–5
1–3
1–8
2
6–9
7.5
+
7–9
7.5
2
6–8
7
+
+
2
2
+
2
+
2
2
+
2
+
2
+
+
2
+
2
+
2
+
2
+
2
2
2
+
2
2
2
+
+
+
+
+
2
2
+
2
2
2
2
2
2
2
2
2
2
2
+
2
+
2
+
2
+
2
+
+
2
+
+
+
+
2
+
2
+
2
+
R
S
S
R
R
S
W
+
W
+
2
+
+
+
W
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Winogradskyella psychrotolerans sp. nov.
Table 1. cont.
Characteristic
1
2
3
Cefoperazone (75)
Cefuroxime (30)
Cephotaxime (30)
Penicillin-G
(10 units/disc)
Cefazolin (30)
Lincomycin (15)
Kanamycin (30)
Chemotaxonomic
characteristics:
Major fatty acids
R
R
S
R
R
R
R
S
S
S
R
S
S
R
R
S
S
R
R
R
S
iso-C13 : 0, iso-C14 : 0, iso-C15 : 0,
iso-C15 : 0, anteiso-C15 : 0, iso-C15 : 1 G,
iso-C15 : 0, anteiso-C15 : 0,
iso-C15 : 1 G, anteiso-C15 : 1 A, anteiso-C15 : 0, iso-C15 : 1 G, anteiso-C15 : 1 A, anteiso-C15 : 1 A, iso-C15 : 0 3-OH,
iso-C15 : 0 3-OH
iso-C15 : 0 3-OH, iso-C17 : 0 3-OH
summed feature 9
Polar lipids
PE, AL1, AL2, L1, L3
PE, AL1, AL2, L1
PE, AL1, L1, L2
Menaquinones
MK-6, MK-7
MK-6
MK-6
DNA G+C content (mol%)
34.7±1.2
35.5±1.2
33.1±1.2
RS-3T clustered with the species of the genus Winogradskyella
and clustered with W. pacifica and together with W.
thalassocola (Fig. 1). The topology of the NJ tree was
essentially the same (Fig. S1 available in IJSEM Online).
Hence, the type strains of W. pacifica and W. thalassocola
were grown under the same conditions as strain RS-3T and
used as reference strains in most phenotypic and chemotaxonomic tests.
Cell morphology and motility were studied using a light
microscope. Growth at different temperatures (4, 10, 18,
30, 37 and 40 uC) were ascertained using MA agar medium
and salt tolerance [0 (without NaCl), 1, 2, 3, 4, 5, 6, 8 and
10 % (w/v)] were ascertained using nutrient agar (NA,
HIMEDIA) medium. Growth at pH 5, 6, 7, 7.5, 8, 8.5, 9,
9.5, 10, 11 and 12 was assessed on MA buffered with citric
acid–NaOH (for pH 5 and 6), NaHPO4–Na2HPO4 (for
pH 7 and 8), glycine–NaOH (for pH 9 and 10) or Tris/HCl
or NaOH (for pH 11 and 12). Biochemical characteristics
such as activity of oxidase, catalase, lysine decarboxylase,
ornithine decarboxylase, nitrate reduction, hydrolysis of
aesculin, gelatin, ONPG, starch, Tween 60, carbon source
assimilation, H2S production and the sensitivity to 17
different antibiotics using the disc diffusion method with
commercially available discs (HIMEDIA) were determined
by previously described methods (Lányı́, 1987; Smibert &
Krieg, 1994). Biochemical characteristics were also assessed
using the Hi25 Enterobacteriaceae identification kit and the
HiCarbohydrate kit parts A, B and C (HIMEDIA) according to the manufacturer’s protocol. The phenotypic characteristics of strain RS-3T are listed in the species description
and in Table 1.
Standardization of the physiological age of strains RS-3T,
W. pacifica KCTC 22997T and W. thalassocola DSM 15363T
was done based on the protocol (http://www.microbialid.
com/PDF/TechNote_101.pdf) given by Sherlock Microbial
Identification System (MIDI). For cellular fatty acids
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analysis, strains RS-3T, W. pacifica KCTC 22997T and W.
thalassocola DSM 15363T were grown on MA plates at
18 uC for 4, 3 and 5 days. Cellular fatty acid methyl esters
(FAMEs) were obtained from cells by saponification, methylation and extraction following the protocol of MIDI.
Cellular FAMEs were separated by GC (6850) and identified
and quantified with MIDI System software (version. 6.0,
using the aerobe RTSBA6 method and RTSBA6 database).
Menaquinones and polar lipids were determined in freezedried cells of strain RS-3T and the reference strains. Menaquinones were extracted as described by Collins et al.
(1977) and were analysed by HPLC (Groth et al., 1997).
Polar lipids were extracted and analysed according to the
method described by Komagata & Suzuki (1987). The DNA
of strains RS-3T, W. pacifica KCTC 22997T and W. thalassocola DSM 15363T was isolated according to the procedure of
Marmur (1961) and the G+C content was determined from
melting point (Tm) curves (Sly et al., 1986) obtained by
using a Lambda 2 UV-Vis spectrophotometer (Perkin
Elmer) equipped with the Templab 2.0 software package
(Perkin Elmer). Escherichia coli strain DH5-a was used as a
standard. DNA–DNA hybridization was performed by the
membrane filter method (Tourova & Antonov, 1988) as
described previously (Shivaji et al., 1992; Reddy et al., 2000).
The cellular fatty acid composition of strain RS-3T showed
a pronounced dominance of the branched and hydroxyl
medium chain saturated and unsaturated fatty acids
iso-C15 : 0 (18.0), anteiso-C15 : 0 (16.8), iso-C15 : 1 G (14.2),
anteiso-C15 : 1 A (6.0) and iso-C15 : 0 3-OH (6.8) (Table 2).
The quinones of RS-3T consisted of MK-6 (72.42 %) and
MK-7 (27.58 %). MK-6 was present in reference strains W.
pacifica KCTC 22997T and W. thalassocola DSM 15363T
(Table 1). Strain RS-3T contained phosphatidylethanolamine (PE), two unidentified aminolipids (AL1 and AL2)
and two unidentified lipids (L1 and L3) as total polar lipids
(Fig. 2a). All three strains possessed PE, one unidentified
aminolipid (AL1) and one unidentified lipid (L1) and
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Z. Begum and others
Table 2. Comparison of the fatty acid composition of
Winogradskyella psychrotolerans RS-3T from the closely
related species of the genus Winogradskyella
Strains: 1, Winogradskyella psychrotolerans RS-3T; 2, Winogradskyella
pacifica KCTC 22997T; 3, Winogradskyella thalassocola DSM 15363T.
Results are presented as percentages of the total fatty acids. Fatty acids
amounting to 5 % or more of the total fatty acids are indicated by
bold type. Data for both the taxa are from the present study. ND, not
detected. The strains were grown on MA plates at 186C. Fatty acids
amounting ,1 % in all strains were not considered.
Fatty acid
Saturated
C14 : 0
C16 : 0
C17 : 0
C20 : 0
Unsaturated
C15 : 1v8c
C15 : 1v6c
Branched saturated
iso-C13 : 0
anteiso-C13 : 0
iso-C14 : 0
anteiso-C14 : 0
iso-C15 : 0
anteiso-C15 : 0
Branched unsaturated
iso-C15 : 1 G
anteiso-C15 : 1 A
anteiso-C17 : 1v9c
Hydroxy
iso-C14 : 0 3-OH
C15 : 0 2-OH
C15 : 0 3-OH
iso-C15 : 0 3-OH
C16 : 0 3-OH
iso-C16 : 0 3-OH
C17 : 0 2-OH
iso-C17 : 0 3-OH
Summed feature*
3
9
1
2
3
Description of Winogradskyella psychrotolerans
sp. nov.
0.8
1.7
2.1
1.2
1.0
ND
0.6
2.2
Winogradskyella psychrotolerans (psy.chro.to9le.rans. Gr. adj.
psychros cold; L. part. adj. tolerans tolerating; N.L. part. adj.
psychrotolerans tolerating cold temperature).
ND
ND
ND
ND
1.9
ND
ND
ND
1.9
1.9
2.2
1.2
3.5
3.1
18.0
16.8
8.9
1.9
1.0
4.0
3.2
17.7
17.0
14.2
6.0
1.6
11.6
6.3
ND
1.0
3.2
1.7
8.1
4.5
2.0
ND
6.8
0.3
1.7
1.6
4.4
ND
4.7
ND
6.9
6.0
8.4
9.0
ND
13.7
6.1
1.5
ND
1.8
ND
6.8
0.4
ND
ND
2.0
6.4
1.4
4.1
2.4
1.3
5.6
ND
*Summed features are groups of two or three fatty acids that cannot
be separated by the MIDI system. Summed feature 3 comprises isoC15 : 0 2-OH and/or C16 : 1v7c and/or C16 : 1v6c; summed feature 9
comprises C16 : 0 10-methyl and/or iso-C17 : 1v9c.
differed from each other in presence or absence of one
unidentified aminolipid (AL2) and two unidentified lipids
(L2 and L3) (Fig. 2). DNA–DNA hybridization with W.
pacifica KCTC 22997T and W. thalassocola DSM 15363T
showed a relatedness of 22 % and 42.5 % with respect to strain
RS-3T. This value is lower than 70 %, confirming that the
novel strain RS-3T is not a member of W. pacifica KCTC
22997T and W. thalassocola DSM 15363T (Wayne et al., 1987).
1650
The main features of the strain RS-3T are in line with the
original and emended descriptions of the genus Winogradskyella but it could be distinguished from the closely
related species W. pacifica KCTC 22997T and W. thalassocola
DSM 15363T (Tables 1 and 2). Thus, the cumulative
differences that strain RS-3T exhibits from the closely related
type strains unambiguously supports the creation of a new
species of the genus Winogradskyella for which the name
Winogradskyella psychrotolerans sp. nov. is proposed.
Cells are Gram-negative, non-motile, rod–coccus shaped
(0.4–0.6 mm in diameter and 0.5–1.0 mm in length) and
occur singly. Colonies are round, 1 mm in diameter,
smooth, shiny, mucoid, yellowish, opaque, flat and entire.
Cells grow at 4 to 30 uC and exhibit optimum growth at
18 uC. Requires NaCl for growth and tolerates up to 6 %
(w/v) NaCl. Grows at pH 6–9, with optimum growth at
pH 7.5. Catalase and oxidase activities are present but
ornithine decarboxylase, lysine decarboxylase b-galactosidase and phenylalanine deaminase activities are absent. The
methyl red and Voges–Proskauer’s reactions are negative.
There is no indole production from tryptophan. Nitrate is
reduced and H2S is not produced. Casein, aesculin, gelatin
and urea are hydrolysed but not agar, DNA, starch and
Tween 60. Acid is produced from sucrose, lactose, melibiose,
mannose and mannitol (in the Hi25 Enterobacteriaceae
identification kit) and the new species is able to ferment the
same sugars plus citrate and malonate when using the
HiCarbohydrate kit. Utilizes phenylalanine, isoleucine,
tryptophan, methionine, glutamic acid, cysteine, histidine
and valine but does not assimilate arabinose, xylose,
adonitol, rhamnose, cellobiose, raffinose, trehalose, glucose,
maltose, fructose, galactose, L-arabinose, sucrose, inulin,
sodium gluconate, glycerol, salicin, glucosamine, dulcitol,
inositol, sorbitol, methyl a-D-glucoside, ribose, melezitose,
methyl-a-D-mannoside, xylitol, D-arabinose, sorbose, aspartic acid, alanine and leucine. Susceptible to (mg/disc unless
indicated) norfloxacin (10), lomefloxacin (30), roxithromycin (30), penicillin G (10 units/disc), cefazolin (30), lincomycin (2), chloramphenicol (30), erythromycin (15) and
tetracycline (30); resistant to co-trimoxazole (25), nalidixic
acid (30), colistin (10), ampicillin (10), cefoperazone (75),
cefuroxime (30), cefotaxime (30) and kanamycin (30). The
polar lipids consist of phosphatidylethanolamine (PE), two
unidentified aminolipids (AL1 and AL2) and two unidentified lipids (L1 and L3). The isoprenoid quinones present
are MK-6 and MK-7. The cellular fatty acids composition is
as in Table 2.
The type strain is RS-3T (5CIP 110154T5NBRC 106169T),
isolated from a sediment sample collected from the marine
transect of Kongsfjorden, Ny-Ålesend, Svalbard, Arctic.
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Winogradskyella psychrotolerans sp. nov.
Fig. 2. Two-dimensional thin-layer chromatogram of the total lipids of Winogradskyella psychrotolerans RS-3T (a)
Winogradskyella pacifica KCTC 22997T (b) and Winogradskyella thalassocola DSM 15363T (c) after spraying with
molybdatophosphoric acid. PE, phosphatidylethanolamine; AL1–AL2, unidentified aminolipids; L1–L3, unidentified lipids. The
lipids were identified as phospholipids or aminolipids by spraying with molybdenum blue and ninhydrin reagents respectively.
The genomic DNA G+C content of the type strain is
34.7±1.2 mol%.
Emended description of the genus
Winogradskyella
The description of the genus is as given previously (Ivanova
et al., 2010; Nedashkovskaya et al., 2005, 2012; Yoon et al.,
2011), with the following amendments: cells are both rodand coccus-shaped.
Ivanova, E. P., Christen, R., Gorshkova, N. M., Zhukova, N. V.,
Kurilenko, V. V., Crawford, R. J. & Mikhailov, V. V. (2010).
Winogradskyella exilis sp. nov., isolated from the starfish Stellaster
equestris, and emended description of the genus Winogradskyella. Int J
Syst Evol Microbiol 60, 1577–1580.
Kim, S. B. & Nedashkovskaya, O. I. (2010). Winogradskyella pacifica
sp. nov., a marine bacterium of the family Flavobacteriaceae. Int J Syst
Evol Microbiol 60, 1948–1951.
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.
Komagata, K. & Suzuki, K. (1987). Lipid and cell-wall analysis in
bacterial systematics. Methods Microbiol 19, 161–207.
Acknowledgements
Lane, D. J. (1991). 16S/23S rRNA sequencing. In Nucleic Acid Techniques
in Bacterial Systematics, pp. 115–175. Edited by E. Stackebrandt &
M. Good Fellow. Chichester: Wiley.
S. S. is thankful to the National Centre for Antarctic and Ocean
Research, Goa, and the Council of Scientific and Industrial Research
(CSIR) Network Project on Exploitation of India’s rich microbial
diversity (NWP0006) for funding. T. N. R. S. acknowledges the CSIR,
Government of India for the award of a Research Associateship.
Lányı́, B. (1987). Classical and rapid identification methods for
References
Winogradskyella poriferorum sp. nov., a novel member of the family
Flavobacteriaceae isolated from a sponge in the Bahamas. Int J Syst
Evol Microbiol 55, 1589–1592.
Altschul, S. F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J.
(1990). Basic local alignment search tool. J Mol Biol 215, 403–410.
Collins, M. D., Pirouz, T., Goodfellow, M. & Minnikin, D. E. (1977).
medically important bacteria. Methods Microbiol 19, 1–67.
Lau, S. C. K., Tsoi, M. M. Y., Li, X., Plakhotnikova, I., Dobretsov, S.,
Lau, K. W. K., Wu, M., Wong, P. K., Pawlik, J. R. & Qian, P. Y. (2005).
Marmur, J. (1961). A procedure for the isolation of deoxyribonucleic
acid from microorganisms. J Mol Biol 3, 208–218.
Distribution of menaquinones in Actinomycetes and Corynebacteria. J
Gen Microbiol 100, 221–230.
Nedashkovskaya, O. I., Kim, S. B., Han, S. K., Snauwaert, C.,
Vancanneyt, M., Swings, J., Kim, K. O., Lysenko, A. M., Rohde, M. &
other authors (2005). Winogradskyella thalassocola gen. nov., sp.
Felsenstein, J. (1993).
PHYLIP (phylogeny inference package) version
3.5.1. Distributed by the author. Department of Genome Sciences,
University of Washington, Seattle, USA.
nov., Winogradskyella epiphytica sp. nov. and Winogradskyella eximia
sp. nov., marine bacteria of the family Flavobacteriaceae. Int J Syst Evol
Microbiol 55, 49–55.
Groth, I., Schumann, P., Rainey, F. A., Martin, K., Schuetze, B. &
Augsten, K. (1997). Demetria terragena sp. nov., a new genus of
Nedashkovskaya, O. I., Vancanneyt, M., Kim, S. B. & Zhukova, N. V.
(2009). Winogradskyella echinorum sp. nov., a marine bacterium of the
Actinomycetes isolated from compost soil. Int J Syst Bacteriol 47,
1129–1133.
family Flavobacteriaceae isolated from the sea urchin Strongylocentrotus
intermedius. Int J Syst Evol Microbiol 59, 1465–1468.
Guindon, S., Lethiec, F., Duroux, P. & Gascuel, O. (2005). PHYML
Nedashkovskaya, O. I., Kukhlevskiy, A. D. & Zhukova, N. V. (2012).
Online–a web server for fast maximum likelihood-based phylogenetic
inference. Nucleic Acids Res 33 (Web Server issue, ), W557–W559.
Winogradskyella ulvae sp. nov., a novel epiphyte of a Pacific seaweed
and emended descriptions of the genus Winogradskyella and
http://ijs.sgmjournals.org
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 23:30:58
1651
Z. Begum and others
Winogradskyella thalassocola, Winogradskyella echinorum, Winogradskyella exilis and Winogradskyella eximia. Int J Syst Evol Microbiol 62,
1450–1456.
Pinhassi, J., Nedashkovskaya, O. I., Hagström, Å. & Vancanneyt, M.
(2009). Winogradskyella rapida sp. nov., isolated from protein-
enriched seawater. Int J Syst Evol Microbiol 59, 2180–2184.
Reddy, G. S. N., Aggarwal, R. K., Matsumoto, G. I. & Shivaji, S.
(2000). Arthrobacter flavus sp. nov., a psychrophilic bacterium
Smibert, R. M. & Krieg, N. R. (1994). Phenotypic characterization. In
Methods for General and Molecular Bacteriology, pp. 607–654. Edited
by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg.
Washington, DC: American Society for Microbiology.
Srinivas, T. N. R., Nageswara Rao, S. S. S., Vishnu Vardhan Reddy,
P., Pratibha, M. S., Sailaja, B., Kavya, B., Hara Kishore, K., Begum, Z.,
Singh, S. M. & Shivaji, S. (2009). Bacterial diversity and bioprospect-
isolated from a pond in McMurdo Dry Valley, Antarctica. Int J Syst
Evol Microbiol 50, 1553–1561.
ing for cold-active lipases, amylases and proteases, from culturable
bacteria of Kongsfjorden and Ny-alesund, Svalbard, Arctic. Curr
Microbiol 59, 537–547.
Romanenko, L. A., Tanaka, N., Frolova, G. M. & Mikhailov, V. V.
(2009). Winogradskyella arenosi sp. nov., a member of the family
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., Higgins,
D. G. & Higgins, D. G. (1997). The CLUSTAL_X windows interface:
Flavobacteriaceae isolated from marine sediments from the Sea of
Japan. Int J Syst Evol Microbiol 59, 1443–1446.
flexible strategies for multiple sequence alignment aided by quality
analysis tools. Nucleic Acids Res 25, 4876–4882.
Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new
method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–
425.
Tourova, T. P. & Antonov, A. S. (1988). Identification of microorgan-
Shivaji, S., Ray, M. K., Shyamala Rao, N., Saisree, L., Jagannadham,
M. V., Seshu Kumar, G., Reddy, G. S. N. & Bhargava, P. M. (1992).
Sphingobacterium antarcticus sp. nov.: a psychrotrophic bacterium
from the soils of Schirmacher Oasis, Antarctica. Int J Syst Bacteriol 42,
102–106.
isms by rapid DNA–DNA hybridisation. Methods Microbiol 19, 333–
355.
Wayne, L. G., Brenner, D. J., Colwell, R. R., Grimont, P. A. D., Kandler,
O., Krichevsky, M. I., Moore, L. H., Moore, W. E. C., Murray, R. G. E. &
other authors (1987). International Committee on Systematic
Bacteriology. Report of the ad hoc committee on reconciliation of
approaches to bacterial systematics. Int J Syst Bacteriol 37, 463–464.
Sly, L. I., Blackall, L. L., Kraat, P. C., Tian-Shen, T. & Sangkhobol, V.
(1986). The use of second derivative plots for the determination of
Yoon, B. J., Byun, H. D., Kim, J. Y., Lee, D. H., Kahng, H. Y. & Oh, D. C.
(2011). Winogradskyella lutea sp. nov., isolated from seawater, and
mol% guanine plus cytosine of DNA by the thermal denaturation
method. J Microbiol Methods 5, 139–156.
emended description of the genus Winogradskyella. Int J Syst Evol
Microbiol 61, 1539–1543.
1652
Downloaded from www.microbiologyresearch.org by
International Journal of Systematic and Evolutionary Microbiology 63
IP: 88.99.165.207
On: Fri, 16 Jun 2017 23:30:58