International Journal of Systematic and Evolutionary Microbiology (2002), 52, 823–829
DOI : 10.1099/ijs.0.02008-0
The search for synonyms among
streptomycetes by using SDS-PAGE of wholecell proteins. Emendation of the species
Streptomyces aurantiacus, Streptomyces cacaoi
subsp. cacaoi, Streptomyces caeruleus and
Streptomyces violaceus
1
BCCMTM/LMG Bacteria
Collection, K. L.
Ledeganckstraat 35, 9000
Gent, Belgium
B. Lanoot,1 M. Vancanneyt,1 I. Cleenwerck,1 L. Wang,2 W. Li,2 Z. Liu2
and J. Swings1
2
Institute of Microbiology,
Chinese Academy of
Sciences, Beijing 100080,
P.R. China
Author for correspondence : B. Lanoot. Tel : j32 9 264 5102. Fax : j32 9 264 5092.
e-mail : Benjamin.Lanoot!rug.ac.be
A collection of 93 Streptomyces reference strains were investigated using SDSPAGE of whole-cell proteins. Computer-assisted numerical analysis revealed 24
clusters encompassing strains with very similar protein profiles. Five of them
grouped several type strains with visually identical patterns. DNA–DNA
hybridizations revealed homology values higher than 70 % among these
type strains. According to the current species concept, it is proposed that
Streptomyces albosporeus subsp. albosporeus LMG 19403T is considered as a
subjective synonym of Streptomyces aurantiacus LMG 19358T, that
Streptomyces aminophilus LMG 19319T is considered as a subjective synonym
of Streptomyces cacaoi subsp. cacaoi LMG 19320T, that Streptomyces niveus
LMG 19395T and Streptomyces spheroides LMG 19392T are considered as
subjective synonyms of Streptomyces caeruleus LMG 19399T, and that
Streptomyces violatus LMG 19397T is considered as a subjective synonym of
Streptomyces violaceus LMG 19360T.
Keywords : Streptomyces, SDS-PAGE of whole-cell proteins, synonyms, taxonomy
INTRODUCTION
At the introduction of the genus in 1943 (Waksman &
Henrici, 1943) classifications were based solely on
morphology and pigmentation. The lack of standardized procedures for describing streptomycetes,
together with the unavailability of type strains,
led to species which were poorly described. Moreover, antibiotics producing streptomycetes were
described on the basis of trivial differences in morphology, for patent purposes. This unsound classification system has been improved by the International
Streptomyces Project (ISP) in 1964. Standard procedures for describing streptomycetes were developed
and applied to 463 Streptomyces and (former) Streptoverticillium species. Type strains became publicly
available when they were deposited in four collections.
.................................................................................................................................................
Abbreviation : ISP, International Streptomyces Project.
No attempts were undertaken by the ISP to determine
synonymies among morphologically defined species.
With the introduction of the principles of numerical
taxonomy by Locci et al. (1981) and Williams et al.
(1983), another view on the taxonomy of this complex
genus emerged. Numerical taxonomy of phenotypic
features without heavy emphasis on morphology and
pigmentation allowed a more reliable classification
than those previously defined. The taxonomic study of
Williams et al. (1983) is therefore considered to be a
milestone in Streptomyces systematics. An extensive
species reduction was proposed, and probability
matrices were created to identify isolates (Williams et
al., 1989). Ka$ mpfer & Kroppenstedt (1991) updated
the classification system of Williams et al. (1983) by
extending the numbers of phenotypic traits and by
including more taxa.
Streptomyces systematics is gradually evolving from a
phenotypically dominated taxonomy to a more poly-
02008 # 2002 IUMS Printed in Great Britain
823
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
B. Lanoot and others
.................................................................................................................................................................................................................................................................................................................
Fig. 1. Dendrogram derived from an unweighted pair group method with arithmetic averages (UPGMA) algorithm of r
values (expressed as percentages) of protein profiles. Clusters are delineated at a similarity level of 90 %. Symbols : *,
cultivated for 48 h ; †, growth temperature 48 mC.
824
International Journal of Systematic and Evolutionary Microbiology 52
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
Synonymous species in the genus Streptomyces
phasic taxonomy comprising chemotaxonomic and
genomic information. The large number of validly
described Streptomyces species (more than 500) remains a major obstacle in the setting up of large
fundamental taxonomic studies, including DNA–
DNA hybridizations, and in establishing the link
between phenotypic and genomic information. Furthermore, because of the lack of minimal standards for
defining Streptomyces species (Manfio et al., 1994), the
polyphasic approach remains difficult.
The aim of this study was to screen for synonymous taxa in a representative subset of the genus
Streptomyces, using SDS-PAGE of whole-cell
proteins, and to compare the data with these of
Williams et al. (1989) and Ka$ mpfer & Kroppenstedt
(1991). It has been shown that under highly
standardized cultivation conditions, SDS-PAGE of
whole-cell proteins is an excellent and generally
applicable method for defining relationships at the
species level among large numbers of strains
(Vandamme et al., 1996 ; Descheemaeker et al., 1994).
Manchester et al. (1990) has already demonstrated the
usefulness of SDS-PAGE of whole-cell proteins in
streptomycete systematics.
METHODS
Bacterial strains and cultivation conditions. The strains used
in this study can be found in Fig. 1.
All strains were grown on Bennett’s agar slopes (using
0n01 M phosphate buffer, pH 7n0) and incubated for 24 h at
28 mC unless indicated otherwise. Mycelial suspensions were
prepared in 2 ml 0n1 M phosphate buffer (pH 7n0) and added
to 250 ml flasks containing 80 ml of Bennett’s broth (0n01 M
phosphate buffer, pH 7n0). The flasks were shaken for 24 h at
28 mC unless indicated otherwise. The same cultivation
method was used for preparing protein or DNA extracts.
To determine the influence of incubation time on the
resolution of protein profiles, protein extracts were prepared
from Streptomyces griseus subsp. griseus LMG 5974 and
Streptomyces rimosus LMG 5984T after 18, 36 and 51 h.
Furthermore, the influence of the cultivation method (agar
versus broth) was investigated.
SDS-PAGE of whole-cell proteins. Protein extracts were
prepared according to the protocol of Manchester et al.
(1990), with some modifications as indicated above.
SDS-PAGE of the whole-cell proteins, scanning, and
normalization of electrophoretic patterns were performed as
described by Pot et al. (1994). The similarity between all
traces was calculated using the Pearson product moment
correlation coefficient with the software package 
version 4.2 (Applied Maths). Out of the obtained similarity
matrix, a dendrogram was constructed using the unweighted
pair group method with arithmetic averages (UPGMA)
algorithm.
DNA–DNA hybridizations. DNA of 250–500 mg cells was
prepared using the method of Pitcher et al. (1989)
supplemented with a lysozyme (Serva) step (10 mg ml−" in
1iTE buffer). DNA was finally dissolved in 0.1iSSC. Only
high-molecular-mass DNA was retained for further applications. Microplate hybridizations were performed using the
microplate method described by Ezaki et al. (1989), using
a model FL-2575 microplate reader (Towa Scientific)
for the fluorescence measurements and black Maxisorp
FluoroNunc microplates (Nunc A\S). Biotinylated probe
DNA was sheared by ultrasonication (30 s, duty cycle 50,
output control 1) using a Branson Sonifier (model 250)
equipped with microcup horn and was then hybridized with
single-stranded unlabelled DNA, non-covalently bound to
microplate wells. Hybridizations were performed at 52 mC
in a hybridization solution containing 50 % formamide.
Salmon-sperm DNA was used as the negative control in all
experiments.
Determination of mol % GjC. The direct high-performance
liquid chromatography method of Tamaoka & Komagata
(1984) was used. DNA was hydrolysed into nucleosides with
nuclease P1 and bacterial alkaline phosphatase (Sigma) ; this
was followed by separation using reversed-phase HPLC
(Waters).
RESULTS AND DISCUSSION
SDS-PAGE of whole-cell proteins
Before routine application of SDS-PAGE of wholecell proteins, the method was optimized using
Streptomyces LMG 5974 and LMG 5984T. The extent
to which factors such as cultivation time and medium
(agar versus broth) influence the resolution of protein
profiles was investigated. Our results indicated that
high-resolution and reproducible protein patterns were
obtained only from cultures grown in liquid medium,
whereas cultures grown on solid media yielded a high
background (due to proteinase activity) and consequently gave a lower resolution. It was also demonstrated that the growth phase of cells in liquid medium
should preferably be within the exponential phase,
which means approx. 36 h for fast-growing streptomycetes. Protein patterns of strains harvested at the
early stationary phase (51 h), in conditions analogous
to those used in the study of Manchester et al. (1990),
clearly had a lower resolution. Duplicate protein
extracts were prepared in order to verify the reproducibility of the growth conditions and preparation of
extracts. A correlation value above 92 % was observed
between duplicate protein patterns loaded onto separate gels.
Using the above-mentioned growth conditions, protein patterns were generated from 93 Streptomyces
strains (see Fig. 1) belonging to different phenotypically defined clusters. After processing of the data,
a cluster analysis was performed (Fig. 1). A 90 %
similarity level was used as guideline for delineating 24
clusters (Fig. 1) encompassing strains with very similar
protein profiles. A correlation level (r) of 0n90 or higher
in protein patterns of bacterial strains has often been
used as an indication of possible relatedness at the
species level (Kersters, 1985 ; Manchester et al., 1990).
http://ijs.sgmjournals.org
825
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
B. Lanoot and others
Cluster
LMG 19358T
LMG 19403T
1
LMG 19320T
LMG 19319T
5
LMG 19393T
LMG 19335T
LMG 19360T
LMG 19397T
LMG 19399T
LMG 19395T
8
12
19
LMG 19392T
LMG 19301T
LMG 19327T
LMG 19302T
22
Assignment according to:
Williams et al.
(1989)
Kämpfer & Kroppenstedt
(1991)
Cat. II-sp.13
12
Cat. IV-sp.1
Cat. I-sp.9
63
31
Cat. I-sp.9
31
Cat. I-sp.16
41
Cat. I-sp.16
56
Cat. I-sp.5
9
Cat. I-sp.11
Cat. IV-sp.7
Cat. I-sp.2
50
58
43
Cat. I-sp.2
40
Cat. I-sp.2
47
Cat. I-sp.20
Cat. I-sp.2
1-3
1-3
.....................................................................................................
Fig. 2. All strains found in this study
possessing visually identical protein profiles,
except for cluster 22, encompassing strains
with similar protein profiles. A correlation
is made with the numerical phenotypic
studies of Williams et al. (1989) and
Ka$ mpfer & Kroppenstedt (1991). Molecular
mass markers (from left to right), in kDa :
β-galactosidase (116), bovine albumin
(66), egg albumin (45), glyceraldehyde-3phosphate dehydrogenase (36), carbonic
anhydrase (29), trypsinogen (24), trypsin
inhibitor (21n1) and lysozyme (14n2).
Markers
Table 1. DNA hybridization between several Streptomyces taxa
.................................................................................................................................................................................................................................................................................................................
1, S. albosporeus albosporeus LMG 19403T ; 2, S. aurantiacus LMG 19358T ; 3, S. cacaoi cacaoi LMG 19320T ; 4, S. aminophilus
LMG 19319T ; 5, S. violaceus LMG 19360T ; 6, S. violatus LMG 19397T ; 7, S. spheroides LMG 19392T ; 8, S. niveus LMG 19395T ;
9, S. caeruleus LMG 19399T ; 10, S. griseus griseus LMG 19302T ; 11, S. microflavus LMG 19327T ; 12, S. anulatus LMG 19301T.
Values are meansp ; –, not determined.
SDS-PAGE
cluster
1
5
12
19
22
Name as
received/Strain
1
2
3
4
5
6
7
8
9
10
11
12
DNA relatedness ( %) to :
1
2
3
4
5
6
7
8
9
10
11
12
100
74(7)
–
–
–
–
–
–
–
–
–
–
–
100
–
–
–
–
–
–
–
–
–
–
–
–
100
100(6)
–
–
–
–
–
–
–
–
–
–
–
100
–
–
–
–
–
–
–
–
–
–
–
–
100
90(9)
–
–
–
–
–
–
–
–
–
–
–
100
–
–
–
–
–
–
–
–
–
–
–
–
100
77(4)
85(2)
–
–
–
–
–
–
–
–
–
–
100
100(5)
–
–
–
–
–
–
–
–
–
–
–
100
–
–
–
–
–
–
–
–
–
–
–
–
100
43(2)
61(6)
–
–
–
–
–
–
–
–
–
–
100
45(12)
–
–
–
–
–
–
–
–
–
–
–
100
DNA–DNA hybridization experiments
In the present study, delineated clusters contain not
strains of a single species, but several type strains of
different validly described Streptomyces species. Five
clusters (1, 5, 8, 12 and 19 ; Fig. 2) contain type strains
with identical profiles (there were only small differences
in the intensities of bands). Other type strains grouping
in the respective clusters, and all members of the other
clusters, show highly similar (though not identical)
patterns (there were minor differences in the presence
or absence of bands). DNA–DNA hybridization
experiments were performed to investigate whether
synonymous names might be found among the species
in these clusters.
The DNA–DNA hybridization results among strains
of the five selected clusters (sharing identical patterns)
is presented in Table 1. Within cluster 1, Streptomyces
826
albosporeus subsp. albosporeus LMG 19403T and
Streptomyces aurantiacus LMG 19358T share a DNA
homology of 74 %. Within cluster 5, Streptomyces
cacaoi subsp. cacaoi LMG 19320T and Streptomyces
aminophilus LMG 19319T are 100 % related. Within
cluster 8, Streptomyces endus LMG 19393T shows a
relatedness of 92 % (data from Labeda & Lyons, 1991)
with respect to Streptomyces hygroscopicus subsp.
hygroscopicus LMG 19335T. Within cluster 12,
Streptomyces violaceus LMG 19360T and Streptomyces
violatus LMG 19397T show a DNA relatedness of
90 %. Within cluster 19, Streptomyces spheroides LMG
19392T shares DNA relatedness of 77 % and 84 %,
respectively, with Streptomyces niveus LMG 19395T
and Streptomyces caeruleus LMG 19399T. S. caeruleus
LMG 19399T is highly related to S. niveus LMG
19395T, as they show a DNA homology of 100 %.
These data confirm that streptomycetes possessing
International Journal of Systematic and Evolutionary Microbiology 52
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
Synonymous species in the genus Streptomyces
visually identical protein patterns are genomically
more than 70 % related and represent the same
genomic species.
In a second step, we investigated the level of DNA
homology among the strains showing minor
differences in protein profiles (r 0n90, but differentiated by the presence or the absence of a few bands).
Three strains belonging to cluster 22 (r 0n93), i.e. S.
anulatus LMG 19301T, S. griseus subsp. griseus LMG
19302T and S. microflavus LMG 19327T, were studied.
As presented in Table 1, S. anulatus LMG 19301T
shows genomic relatedness of 61 and 45 %, respectively, to S. griseus subsp. griseus LMG 19302T and S.
microflavus LMG 19327T. S. griseus subsp. griseus
LMG 19302T and S. microflavus LMG 19327T show a
genomic relatedness of 43 %. These intermediate DNA
homology values confirm that strains possessing highly
similar protein patterns but showing minor qualitative
differences may share DNA homologies below 70 %
and may be considered as distinct taxa. Numerical
analysis of the SDS-PAGE patterns of soluble proteins
seems to be a perfect method for screening for highly
related strains. Visual identity is indispensable if the
relationship at the species level is being determined.
A good correlation exists between the data for SDSPAGE of whole-cell proteins (the present study) and
published DNA-hybridization data (e.g. Labeda &
Lyons, 1991 ; Kim et al., 1999). In the study of Labeda
& Lyons (1991), it was proposed that S. endus be
considered as a subjective synonym of S. hygroscopicus. Both strains have identical protein profiles
(cluster 8 ; Fig. 2), confirming their conclusions. In the
same study, it was observed that the phenotypically
similar species Streptomyces violaceusniger and S.
hygroscopicus show an intermediate DNA hybridization level of 42 %. In our study, both strains
shared highly similar (r l 0n89), though not identical,
protein profiles. Kim et al. (1999) recently clarified the
taxonomic status of thermophilic streptomycetes. For
instance, it was shown that Streptomyces thermoviolaceus and Streptomyces thermovulgaris are distinct
taxa, as they share a low DNA hybridization value of
10 %. This low value is confirmed by different protein
patterns (r l 0n83).
Comparison with phenotypic data
A low correlation is observed between the protein
profiles and the numerical phenotypic data provided
by Williams et al. (1989) and Ka$ mpfer & Kroppenstedt
(1991). We expected that taxa sharing high DNA–
DNA hybridization values would group within the
same phenotypic cluster. Only for the type strains of S.
aminophilus, LMG 19319T, and S. cacaoi subsp. cacaoi,
LMG 19320T, do the observed high relatedness (100 %,
Table 1) correlate with their phenotypic cluster assignment (Fig. 2), as both strains are classified in
speciesgroup Cat. I-sp. 9 according to Williams et al.
(1989), and in cluster 31 according to Ka$ mpfer &
Kroppenstedt (1991). A partial correlation with former
studies was observed in the following case : S. hygroscopicus subsp. hygroscopicus LMG 19335T and S.
endus LMG 19393T (DNA hybridization of 92 %) are
both grouped in speciesgroup Cat. I-sp. 16 by Williams
et al. (1989), but belong to cluster 56 and cluster 41,
respectively, according to Ka$ mpfer & Kroppenstedt
(1991). S. niveus LMG 19395T and S. spheroides LMG
19392T (DNA hybridization of 77 %) belong to the
speciesgroup Cat. I-sp. 2 according to Williams et al.
(1989), but are classified in cluster 43 and cluster 40,
respectively, according to Ka$ mpfer & Kroppenstedt
(1991). S. caeruleus LMG 19399T, which belongs to
the same DNA hybridization group (showing 85
and 100 % DNA hybridization, respectively, to S.
spheroides LMG 19392T and S. niveus LMG 19395T),
is grouped in Cat. IV-sp. 7 by Williams et al. (1989),
and in cluster 58 by Ka$ mpfer & Kroppenstedt (1991).
S. violaceus LMG 19360T and S. violatus LMG 19397T
(sharing 90 % DNA hybridization) are classified in
Cat. I-sp. 5 and Cat. I-sp. 11 according to Williams et
al. (1989), and in clusters 9 and 50 according to
Ka$ mpfer & Kroppenstedt (1991). In one case, no
correlation existed between our data and those of
other authors, i.e. S. aurantiacus LMG 19358T and S.
albosporeus subsp. albosporeus LMG 19403T (having
DNA hybridization of 74 %), are classified in Cat. IIsp. 13 and Cat. IV-sp. 1 according to Williams et al.
(1989), and in clusters 12 and 63 by Ka$ mpfer &
Kroppenstedt (1991). These results clearly confirm
that the phenotypic groupings provided by Williams et
al. (1989) and Ka$ mpfer & Kroppenstedt (1991) do not
always reflect the genomic relationships.
In the genus Streptomyces (conventional) phenotypic
data still dominate the classification system. In other
genera containing fewer species (e.g. the genus
Helicobacter ; Dewhirst et al., 2000), this phenotypic
classification system has already gradually evolved
into a polyphasic approach (in which genomic data are
most often used as a basis for delineation of taxa). It is
logical that we should pursue an analogous polyphasic
taxonomy for the genus Streptomyces, and that it is no
longer acceptable to use different criteria for speciation
in different groups of bacteria. Genotypy should
preferably precede phenotypy. Only when genomebased groups are defined can the search for phenotypic
traits (which may or may not be successful) begin.
From our study, it is clear that SDS-PAGE of wholecell proteins, applied under highly standardized
conditions, is a valuable phenotypic tool for screening
for synonyms within the genus Streptomyces, provided
that DNA–DNA hybridization experiments are performed to confirm genomic relatedness at the species
level.
On the basis of the data from this paper, we propose
the following synonyms in the genus Streptomyces : S.
albosporeus subsp. albosporeus LMG 19403T is a
subjective synonym of S. aurantiacus LMG 19358T ; S.
aminophilus LMG 19319T is a subjective synonym of S.
cacaoi subsp. cacaoi LMG 19320T ; S. niveus LMG
http://ijs.sgmjournals.org
827
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
B. Lanoot and others
19395T and S. spheroides LMG 19392T are subjective
synonyms of S. caeruleus LMG 19399T ; S. violatus
LMG 19397T is a subjective synonym of S. violaceus
LMG 19360T. Emended descriptions are given below
and are based on the species descriptions, as given by
Williams et al. (1989), and the original descriptions.
Emended description of Streptomyces cacaoi subsp.
cacaoi Waksman in Bunting 1932, 515–517 ; Waksman
and Henrici 1948, 951AL
Spore chains are spirales ; the spore surface is smooth.
The aerial mass is in the white-yellow colour series.
The reverse side of a colony has no distinctive
pigments. Melanin pigment is not formed and soluble
pigments are not produced. The following carbohydrates are utilized for growth : -glucose, -arabinose, -xylose, -mannitol and -fructose. There is no
growth, or only traces of growth, on iso-inositol and
rhamnose. The GjC content of the DNA is 73 mol %.
The type strain is IMRU 3082T (l LMG 19320T).
Subjective synonym : Streptomyces aminophilus IJSB
30 : 371AL ATCC 14961T (l LMG 19319T).
Emended description of Streptomyces caeruleus
(based on the original descriptions of S. caeruleus
and S. spheroides) (Baldacci 1944) Pridham,
Hesseltine and Benedict 1958, 60AL (Actinomyces
caeruleus Baldacci 1944, 180)
Spore chains are rectiflexibiles or spirales. The spore
surface is smooth. The aerial mass is yellow or grey.
The reverse side of a colony is dark blue-violet to black
(for S. caeruleus) and is pale yellow to light yellowish
brown (for S. spheroides). Melanoid pigments are not
formed. Bluish-black to greenish-black diffusible
pigments are produced on some media (for S.
caeruleus). Carbon utilization in S. caeruleus occurs
only on -glucose. Very poor growth is observed on
Pridham and Gottlieb carbon-utilization medium
when other carbon sources are being used. S.
spheroides utilizes -glucose, -arabinose, sucrose, xylose, -mannitol, -fructose and rhamnose. Growth
is possible on iso-inositol and raffinose. The GjC
content of the DNA is 71 mol %. The type strain is
IPV 930T (l ISP 5103T l LMG 19399T). Subjective
synonyms : Streptomyces niveus Smith, Dietz, Sokolski
and Savage 1956, IJSB 30 :394AL NRRL 2466 (LMG
19395T) and Streptomyces spheroides Wallick et al.
1956, IJSB 30 : 401AL DSM 40292T (LMG 19392T).
Emended description of Streptomyces aurantiacus
(Rossi-Doria 1891), Waksman in Waksman and
Lechevalier 1953, 53AL Streptothrix aurantiaca
Rossi-Doria 1891, 417
Spore chains are spirales. The spore surface is smooth.
Aerial mycelium (if developed) belongs to the red
colour series. The reverse side of a colony is yellow,
828
yellow-brown, red or red-brown. Melanoid pigments
are not formed and soluble pigments are not produced.
The following carbohydrates are utilized for growth :
-glucose, -arabinose, rhamnose, -xylose, raffinose,
sucrose (only for S. aurantiacus), iso-inositol (only for
S. aurantiacus), -galactose (only for S. albosporeus
subsp. albosporeus), and meso-inositol (only for S.
albosporeus subsp. albosporeus). The GjC content of
the DNA is 70 mol %. The type strain is INMI 1373T
(l LMG 19358T). Subjective synonym : Streptomyces
albosporeus subsp. albosporeus IJSB 30 : 370AL DSM
40795T (LMG 19403T).
Emended description of Streptomyces violaceus
(Rossi-Doria) Waksman and Henrici l Actinomyces
violaceus (Rossi-Doria) Gasperini 1894, emend.
Krasilnikov 1941 ; Streptomyces violaceus (RossiDoria) Waksman in Waksman and Lechevalier 1953,
43
Spore chains are spirales. Spore chains are moderately
long, comprising 10–50 or more spores per chain. The
spore surface is spiny. The aerial mass is in the white or
red colour series. The reverse side of a colony is reddish
or reddish-orange to purple. Reverse mycelium pigment is a pH indicator. Melanoid pigments are formed
(sometimes slowly). A red to violet pigment is found in
the medium in yeast-malt agar, oatmeal agar, saltsstarch agar and glycerol-asparagine agar. This pigment
is pH-sensitive. The following carbohydrates are
utilized for growth : -glucose, -arabinose, -xylose,
-fructose, rhamnose, sucrose, raffinose, iso-inositol
and -mannitol. The GjC content of the DNA is 71
mol %. The type strain is INMI 1T (l LMG 19360T).
Subjective synonym : Streptomyces violatus IJSB
30 : 404AL INMI 1205T (LMG 19397T).
ACKNOWLEDGEMENTS
This study was performed within the framework of the
Belgian–Chinese programme ‘ Identification and classification of actinomycetes, specifically bioactive Streptomyces
strains isolated from Chinese soil ’, supported by OSTC
Belgium (BL\02\C10) and MOST\NSFC P. R. China. We
thank Dirk Dewettinck (RUG, Belgium) for excellent
technical assistance in performing SDS-PAGE of whole-cell
proteins. The authors are also indebted to Professor R. M.
Kroppenstedt (DSMZ, Germany) and Dr T. Kudo (JCM,
Japan) for providing cultures.
REFERENCES
Descheemaeker, P., Pot, B., Ledeboer, A. M., Verrips, T. & Kersters,
K. (1994). Comparison of the Lactococcus lactis differential medium
(DCL) and SDS-PAGE of whole-cell protein extracts for the identification of lactococci to subspecies level. Syst Appl Microbiol 17,
459–466.
Dewhirst, F. E., Fox, J. G. & On, S. L. W. (2000). Recommended
minimal standards for describing new species of the genus Helicobacter.
Int J Syst Evol Microbiol 50, 2231–2237.
Ezaki, T., Hashimoto, Y. & Yabuuchi, E. (1989). Fluorometric
deoxyribonucleic acid–deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in
International Journal of Systematic and Evolutionary Microbiology 52
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07
Synonymous species in the genus Streptomyces
which radioisotopes are used to determine genetic relatedness among
bacterial strains. Int J Syst Bacteriol 39, 224–229.
Ka$ mpfer, P. & Kroppenstedt, R. M. (1991). Probabilistic identification of streptomycetes using miniaturized physiological tests. J Gen
Microbiol 137, 1893–1902.
Kersters, K. (1985). Numerical methods in the classification of bacteria
by protein electrophoresis. In Computer-assisted Bacterial Systematics,
pp. 337–68. Edited by M. Goodfellow, D. Jones & F. G. Priest.
London : Academic Press.
Kim, B., Sahin, N., Minnikin, D. E., Zakrzewska-Czerwinska, J.,
Mordarski, M. & Goodfellow, M. (1999). Classification of thermophilic streptomycetes including the description of Streptomyces thermoalcalitolerans sp. nov. Int J Syst Bacteriol 49, 7–17.
Labeda, D. P. & Lyons, A. J. (1991). The Streptomyces violaceusniger
cluster is heterogeneous in DNA relatedness among strains : emendation
of the descriptions of S. violaceusniger and S. hygroscopicus. Int J Syst
Bacteriol 41, 398–401.
Locci, R., Rogers, J., Sardi, G. & Schofield, G. M. (1981). A
preliminary numerical study on named species of the genus
Streptoverticillium. Anni Microbiol 31, 115–121.
Manchester, L., Pot, B., Kersters, K. & Goodfellow, M. (1990).
Classification of Streptomyces and Streptoverticillium species by numerical analysis of electrophoretic protein patterns. Syst Appl Microbiol
13, 333–337.
Manfio, P. G., Zakrzewska-Czerwinska, J., Atalan, E. & Goodfellow, M. (1994). Towards minimal standards for the description
of Streptomyces species. In Proceedings of the 9th International
Symposium on the Biology of Actinomycetes, Moscow, 1994, pp. 1–14.
Pitcher, D. D., Saunders, N. A. & Owen, R. J. (1989). Rapid
extraction of bacterial genomic DNA with guanidinium thiocyanate.
Lett Appl Microbiol 8, 151–156.
Pot, B., Vandamme, P. & Kersters, K. (1994). Analysis of electrophoretic whole-organism protein fingerprints. In Modern Microbial
Methods. Chemical Methods in Prokaryotic Systematics, pp. 493–521.
Edited by M. Goodfellow & A. G. O ’Donnell. Chichester : Wiley.
Tamaoka, J. & Komagata, K. (1984). Determination of DNA base
composition by reversed-phase high-performance liquid chromatography. FEMS Microbiol Lett 25, 125–128.
Vandamme, P., Pot, B., Gillis, M., De Vos, P., Kersters, K. & Swings,
J. (1996). Polyphasic taxonomy, a consensus approach to bacterial
systematics. Microbiol Rev 60, 407–438.
Waksman, S. & Henrici, A. (1943). The nomenclature and classification of the actinomycetes. J Bacteriol 46, 337–341.
Williams, S. T., Goodfellow, M., Wellington, E. M. H., Vicker, J. C.,
Alderson, G., Sneath, P. H. A. & Sackin, M. J. (1983). Numerical
classification of Streptomyces and related genera. J Gen Microbiol 129,
1743–1813.
Williams, S. T., Goodfellow, M. & Alderson, G. (1989). Streptomyces
and related genera. In Bergey’s Manual of Systematic Bacteriology, vol.
IV, section 29. Edited by S. T. Williams, M. E. Sharpe & J. G. Holt.
Baltimore : Williams & Wilkins.
http://ijs.sgmjournals.org
829
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Thu, 15 Jun 2017 14:50:07