ORIGINAL PAPER

Streptomyces muensis sp. nov.

Debananda S. Ningthoujam • Salam Nimaichand • Dollyca Ningombam •

K. Tamreihao • Li Li • Yong-Guang Zhang • Juan Cheng • Min-Jiao Liu •

Wen-Jun Li

Received: 22 May 2013 / Accepted: 10 September 2013 / Published online: 14 September 2013

� Springer Science+Business Media Dordrecht 2013

Abstract A strain of Streptomyces, MBRL 179T,

isolated from a sample from a Limestone quarry

located at Hundung, Manipur, India, was character-

ized by polyphasic taxonomy. The strain formed a

monophyletic clade with Streptomyces spinoverruco-

sus NBRC 14228T (16S rRNA gene sequence simi-

larity of 99.3 %) in the Neighbour-joining tree. DNA–

DNA hybridization experiment gave a DNA–DNA

relatedness value of 34.7 % between MBRL 179T and

S. spinoverrucosus NBRC 14228T. Strain MBRL 179T

contained LL-diaminopimelic acid, xylose, glucose,

and mannose in the whole cell-wall hydrolysates along

with small amount of ribose. The major polar lipids

detected were diphosphatidylglycerol, phosphatidyl-

ethanolamine, phosphatidylglycerol, phosphatidylin-

ositol and phosphatidylinositolmannoside, with other

unknown phospholipids and aminophospholipid. MK-

9(H6), MK-9(H8) and MK-9(H4) were the predomi-

nant menaquinones detected. The major fatty acids

were anteiso-C16:0 (28.1 %), iso-C16:0 (20.3 %), C16:0

(9.4 %) and anteiso-C17:0 (8.3 %). The G?C content of

the genomic DNA was 71.1 %. Based on the polyphasic

experiment results, the strain MBRL 179T merits recog-

nition as a representative of a novel species of the genus

Streptomyces for which the name Streptomyces muensis

sp. nov. is proposed; the type strain is MBRL 179T (=JCM

17576T = KCTC 29124T).

Keywords Streptomyces muensis sp. nov. �Hundung � Limestone quarry

Electronic supplementary material The online version ofthis article (doi:10.1007/s10482-013-0035-x) contains supple-mentary material, which is available to authorized users.

D. S. Ningthoujam � S. Nimaichand � K. Tamreihao

Microbial Biotechnology Research Laboratory,

Department of Biochemistry, Manipur University,

Canchipur, Imphal 795003, Manipur, India

e-mail: debananda.ningthoujam@gmail.com

D. S. Ningthoujam � D. Ningombam

State Biotech Hub, Department of Biochemistry, Manipur

University, Canchipur, Imphal 795003, Manipur, India

S. Nimaichand � J. Cheng � M.-J. Liu � W.-J. Li (&)

Key Laboratory of Microbial Diversity in Southwest

China, Ministry of Education, Yunnan Institute of

Microbiology, Yunnan University, Kunming 650091,

People’s Republic of China

e-mail: liact@hotmail.com; wjli@ynu.edu.cn

L. Li � Y.-G. Zhang � W.-J. Li

Key Laboratory of Biogeography and Bioresource in Arid

Land, CAS, Xinjiang Institute of Ecology and Geography,

Chinese Academy of Sciences, Ur}umqi 830011, People’s

Republic of China

123

Antonie van Leeuwenhoek (2013) 104:1135–1141

DOI 10.1007/s10482-013-0035-x

Introduction

The genus Streptomyces is characterized by the

presence of high DNA G?C content, formation of

extensively branched substrate and aerial mycelia, the

presence of LL-diaminopimelic acid (LL-DAP) and

absence of characteristic sugars in the cell wall (cell

wall type I) (Kampfer 2012). It is one of most

commonly isolated taxa from soil and has many

biotechnological applications. Two of the common

antifungal compounds produced by this genus are

nystatin (Streptomyces noursei) and amphotericin B

(Streptomyces nodosus), both of which belongs to the

group of polyene antibiotics (Brautaset et al. 2000;

Caffrey et al. 2001). Yuan and Crawford (1995)

reported that a strain of Streptomyces lydicus was a

potential biocontrol agent. Members of the genus have

also been reported to play a role in plant growth

promotion (Hamdali et al. 2008; Khamna et al. 2009).

At the time of writing, there are over 600 Streptomyces

species cited in literature (Euzeby 2013). The present

study presents the polyphasic characterization of a

novel strain, MBRL 179T, which is proposed as

representative of a novel species of the genus

Streptomyces.

Materials and methods

Strain and culture conditions

Strain MBRL 251T was isolated from a soil sample

collected from a limestone quarry at Hundung,

Manipur, India (25.05�N, 94.33�E) on Gauze’s

Medium No. 1 adjusted to pH 5.3 as the selective

isolation medium with the procedure as described

earlier (Nimaichand et al. 2012). Strain MRBL 179T

was preserved as lyophilized spore suspensions in

skimmed milk at room temperature and as glycerol

suspensions (20 %, v/v) at -80 �C.

Phenotypic characterization

To observe its morphological characteristics, strain

MBRL 179T was cultivated aerobically on Gauze’s

Medium No. 1 (28 �C) for 2 weeks. Morphology of

spores and mycelia was observed by using light

microscopy (Axioscope A1, Zeiss) and scanning

electron microscopy (SEM) (JSM-6360, Jeol)

(Williams and Davies 1967). Growth on various

International Streptomyces Project (ISP, Shirling and

Gottlieb 1966) media, tryptic soy agar (TSA, Difco),

Starch Casein Nitrate Agar (SCNA), Gauze’s Medium

No. 1, Czapek’s agar and Nutrient agar (NA) were

observed. The colony color was determined using the

ISCC-NBS color chart (Kelly 1964). Utilization of

sole carbon and nitrogen sources was determined as

described by Shirling and Gottlieb (1966). Tests for

decomposition of casein, and acid production from

carbohydrates were performed following the methods

of Gordon et al. (1974). Hydrolysis of gelatin and

starch was determined as described by Collins et al.

(2004) and Tweens 20, 40, 60 and 80 according to

Sierra (1957). Growth at different temperatures (5, 15,

28, 37, 42, 50 and 60 �C), pH (4, 5, 6, 7, 8, 9 and 10)

and NaCl concentrations (0, 2, 5, 7 and 10 % w/v) were

determined on TSA as described by Goodfellow et al.

(1986). Catalase activity was observed by assessing

bubble production in 3 % (v/v) H2O2 and oxidase

activity was determined by using 1 % (w/v) solution of

tetramethyl-p-phenylenediamine (Kovacs 1956).

Other biochemical tests including Voges-Proskauer,

methyl red and the production of indole were per-

formed as described by Goodfellow et al. (1986).

Chemotaxonomy

The amino acid content of the cell wall was deter-

mined according to Staneck and Roberts (1974) and

the sugars of the whole cell hydrolysates were

analyzed as described by Tang et al. (2009). For other

chemotaxonomic analyses, cell biomass from 2 weeks

old culture on tryptic soy broth (TSB, Difco) was

harvested by centrifugation, washed with distilled

water and lyophilized. Polar lipids were extracted and

analyzed by two-dimensional TLC as described by

Minnikin et al. (1984). The extraction of menaqui-

nones was performed as described by Collins et al.

(1977) and analyzed by HPLC (Tamaoka et al. 1983).

Cellular fatty acids were extracted, methylated and

analyzed by using the Sherlock Microbial Identifica-

tion System (MIDI) according to the method of Sasser

(1990) and the manufacturer’s instructions. The fatty

acid methyl esters were then analysed by GC (Agilent

Technologies 7890A GC System) by using the

Microbial Identification software package (Sherlock

Version 6.1; MIDI database: TSBA6).

1136 Antonie van Leeuwenhoek (2013) 104:1135–1141

123

Molecular analysis

Genomic DNA extraction and PCR amplification of

the 16S rRNA gene was performed as described by Li

et al. (2007) The almost complete 16S rRNA gene

sequence (1479 bp) of the strain was identified using

the EzTaxon-e server database (Kim et al. 2012) and

aligned with the 16S rRNA gene sequences of other

Streptomyces species using CLUSTAL X version 2.1

(Larkin et al. 2007). Phylogenetic analyses were

performed using the software package MEGA version

5.1 (Tamura et al. 2011). Phylogenetic distances were

calculated with the Kimura two-parameter model

(Kimura 1983) and tree topologies were inferred using

the maximum-parsimony (Fitch 1972) and neighbor-

joining (Saitou and Nei 1987) methods. To determine

the support of each clade, bootstrap analysis was

performed with 1,000 resamplings (Felsenstein 1985).

The G?C content of the genomic DNA was deter-

mined according to the method described by Mesbah

et al. (1989). DNA–DNA relatedness was studied in

triplicate by the thermal renaturation method (DeLey

et al. 1970) using Lambda 35 UV/Vis Spectropho-

tometer (Perkin Elmer) equipped with PTP 6 ? 6

Peltier Temperature Programmer (Perkin Elmer) and

BG-chiller E15 (Baygene Biotech Company Limited).

Results and discussion

Strain MBRL 179T formed extensive substrate and

white colour aerial mycelia in Gauze’s Medium No. 1

with spiral spore chains (*15-17 spores, Fig. 1). The

spore surface appears to be smooth or sometimes

shriveled. The strain grew well in all the media tested

(Table 1). MBRL 179T differs from the related type

strain Streptomyces spinoverrucosus NBRC 14228T

Fig. 1 Scanning electron micrographs for strain MBRL 179T

grown on Gauze’s Medium No. 1 for 2 weeks at 28 �C, Bar

2 lm

Table 1 Culture

characteristics of strain MBRL

179T on different ISP and other

selective media as observed

using ISCC-NBS Color Chart

(Kelly, 1964)

Medium Colour of mycelium Soluble pigment

Aerial (Spore mass) Substrate

ISP2 Black Black Orange

ISP3 White Grey –

ISP4 White Yellow Pink –

ISP5 Reddish Brown Black –

ISP6 Grey – Black

ISP7 Reddish Brown Black –

NA Grey Greenish Yellow –

Czapek’s White Yellowish Brown –

SCNA Grey Reddish Brown –

TSA Grey – Orange

Gauze’s Medium No. 1 White Blue Blue

Antonie van Leeuwenhoek (2013) 104:1135–1141 1137

123

(Diab and Al-Gounaim 1982) in the utilization of

adonitol, L-arabinose, cellobiose, fructose, inulin,

maltose, rhamnose and L-tryptophan as sole C/N

sources. Unlike NBRC 14228T, it could not produce

acid from galactose, glucose, lactose and mannose.

The differentiating properties of strain MBRL 179T

from NBRC 14228T are listed in Table 2 and the

detailed phenotypic characteristics are mentioned in

the species description.

Strain MBRL 179T had LL-diaminopimelic acid as

the diagnostic cell wall diamino acid, and xylose

(51.8 %), glucose (26.2 %) and mannose (18.5 %)

were detected in the whole cell hydrolysates along

with small amounts of ribose (3.6 %). The major polar

lipids detected were diphosphatidylglycerol, phospha-

tidylethanolamine, phosphatidylglycerol, phosphati-

dylinositol and phosphatidylinositolmannoside, with

other unknown phospholipids and aminophospholipid

(see Supplementary Fig. S1). MK-9(H6) (49.9 %),

MK-9(H8) (32.4 %) and MK-9(H4) (11.1 %) were the

predominant menaquinones detected along with small

amount of MK-9(H2) (6.6 %). The fatty acid methyl

ester profile ([1 %) contained anteiso-C15:0 (28.1 %),

iso-C16:0 (20.3 %), iso-C15:0 (16.0 %), C16:0 (9.4 %),

anteiso-C17:1 (8.3 %), iso-C17:0 (4.8 %), iso-C14:0 (3.9 %),

Summed Feature 3 containing C16:1x7c and/or

C16:1x6c (3.4 %) and Summed Feature 9 containing

C19:1x11c and/or C19:1x9c (1.2 %).

The G ? C content of the genomic DNA was

71.1 %. The phylogenetic tree of strain MBRL 179T

(Fig. 2) based on neighbor-joining method formed a

monophyletic clade with Streptomyces spinoverruco-

sus NBRC 14228T (16S rRNA gene sequence similar-

ity of 99.3 %; 10 nucleotides difference of total 1454

nucleotides). The phylogenetic position is supported

by other tree-making algorithms and by a bootstrap of

59 % in the neighbor joining tree Hence, the strain S.

spinoverrucosus NBRC 14228T was considered for the

DNA–DNA hybridization studies. The experiments

showed that strain MBRL 179T displayed low DNA–

DNA reassociation values with S. spinoverrucosus

NBRC 14228T (34.7 ± 4.3 %), thereby indicating that

the whole genomic DNA relatedness values are well

below the delineating 70 % cut-off point for species

identification (Wayne et al. 1987).

The chemotaxonomic data and the phylogenetic

analysis clearly indicates the affiliation of the strain

Table 2 Differential characteristics between strain MBRL

179T and S. spinoverrucosus NBRC 14228T

Characteristics S. muensis

MBRL 179T

S. spinoverrucosus

NBRC 14228T

pH range 6–10 6–9

Acid production from

Galactose - ?

Glucose - ?

Lactose - ?

Mannose - ?

Utilization of sole C-sources

Adonitol ? -

L-Arabinose ? -

Cellobiose ? -

Fructose ? -

Galactose - ?

Inositol - ?

Inulin ? -

Maltose ? -

Rhamnose ? -

Sorbitol - ?

Utilization of sole N-sources

L-Glutamine - ?

L-Threonine - ?

L-Tryptophan ? -

Major fatty acids ([ 5 %)

iso-C15:0 16.1 6.6

anteiso-C15:0 28.1 11.9

iso-H C16:1 - 13.1

iso-C16:0 20.3 38.3

C16:0 9.4 -

anteiso-C17:1x9c - 8.3

anteiso-C17:1 8.3 8.4

G ? C mol % 71.1 70.9

Note: ? positive, - negative, w weakly positivem, all the data

were from this study

All the test strains are positive for hydrolysis of Tweens 20, 40, 60,

80, starch; catalase, oxidase and citrate utilization tests, and acid

production from sucrose. All of them utilize lactose, mannitol,

mannose, raffinose, salicin, trehalose, xylose, L-alanine, L-arginine,

L-aspartic acid, L-isoleucine, L-leucine, L-lysine, L-methionine, L-

phenylalanine, L-proline, L-tyrosine and L-valine as sole C and N

sources. They showed negative results for hydrolysis of casein,

gelatin and urea, MR, VP and indole production tests, and acid

production from fructose and maltose. All the above strains could

not utilize dulcitol, melibiose and L-cysteine, as sole C or N

sources. They grew in the temperature range 28–37 �C and tolerate

up to 2 % NaCl

1138 Antonie van Leeuwenhoek (2013) 104:1135–1141

123

MBRL 179T to the genus Streptomyces. The genotypic

and phenotypic features suggest that strain MBRL

179T could be clearly distinguished from its closest

phylogenetic relative. Besides low DNA–DNA relat-

edness with the closest phylogenetic neighbour, the

strain is also distinguished from strain NBRC 14228T

by several phenotypic properties as listed in Table 2.

Therefore, the Hundung strain MBRL 179T is consid-

ered to represent a new species of the genus Strepto-

myces, for which the name Streptomyces muensis sp.

nov. is proposed.

Description of Streptomyces muensis sp. nov.

Streptomyces muensis (mu.en’sis. N.L. masc. adj.

muensis derived from the acronym MU for Manipur

University, Canchipur, India where the type strain has

been isolated).

Gram-stain positive, aerobic and spore chains

containing up to 15–17 spores. Spiral spore chain,

and each spore on maturity measures 0.5–1.5 lm in

diameter. Growth occurs at 28–37 �C and pH 6–10,

with optimum growth at 28 �C and pH 7. Growth

occurs in presence of up to 2 % NaCl. Utilizes

adonitol, L-arabinose, cellobiose, fructose, inulin,

lactose, maltose, mannitol, mannose, raffinose,

rhamnose, salicin, sucrose, trehalose and xylose as

sole carbon sources; and L-alanine, L-arginine, L-

aspartic acid, L-isoleucine, L-leucine, L-lysine, L-

methionine, L-phenylalanine, L-proline, L-tryptophan,

L-tyrosine and L-valine as sole nitrogen sources. Does

not utilize dulcitol, galactose, inositol, melibiose,

sorbitol, L-cysteine, L-glutamine and L-threonine as

either sole carbon or nitrogen sources. Acid produc-

tion from sucrose, but not from fructose, galactose,

glucose, lactose, maltose and mannose. Hydrolyzes

Tweens 20, 40, 60 and 80, and starch, but not casein,

gelatin and urea. Positive in catalase, oxidase and

citrate utilization tests, but negative in methyl red,

Voges-Proskauer and indole production tests. Con-

tains LL-diaminopimelic acid, xylose, glucose and

mannose with small amount of ribose in the cell wall

hydrolysates. MK-9(H6), MK-9(H8) and MK-9(H4)

are the predominant menaquinones present, while the

polar lipids consist of diphosphatidylglycerol, phos-

phatidylethanolamine, phosphatidylglycerol, phos-

phatidylinositol and phosphatidylinositolmannoside,

with other unknown phospholipids, and amino-

phospholipid. The fatty acid profile ([1 %) is as

follows: anteiso-C15:0, iso-C16:0, iso-C15:0, C16:0, an-

teiso-C17:1, iso-C17:0, iso-C14:0, Summed Feature 3

containing C16:1x7c and/or C16:1x6c and Summed

Feature 9 containing C19:1x11c and/or C19:1x9c.

S. violaceus NBRC 13103T (AB184315)

S. roseoviolaceus ISP 5277T (AJ399484)

S. purpurascens NBRC 13077T (AB184859)

S. hawaiiensis NBRC 12784T (AB184143)

S. spinoverrucosus NBRC 14228T (AB184578)

Streptomyces muensis MBRL 179T (JN560155)

S. lusitanus NBRC 13464T (AB184424)

S. coerulescens ISP 5146T (AY999720)

S. bellus ISP 5185T (AJ399476)

S. coeruleorubidus NBRC 12844T (AB184849)

S. lomondensis NBRC 15426T (AB184673)

S. parvulus NBRC 13193T (AB184326)

S. malachitospinus NBRC 101004T (AB249954)

S. iakyrus NBRC 13401T (AB184877)

S. indiaensis NBRC 13964T (AB184553)

99

75

59*

98

65*

0.001

Fig. 2 Neighbour-joining tree based on 16S rRNA gene

sequences, showing the relationships between strain MBRL

179T and other type strains of Streptomyces species. Actinopla-

nes philippinensis IFO 13878T (D85474) was used as the

outgroup. Asterisks indicate branches that were also recovered

using the maximum parsimony tree. Numbers at nodes are levels

of bootstrap support (%) for branch points (1000 resamplings).

Bar, 0.001 substitutions per nucleotide position

Antonie van Leeuwenhoek (2013) 104:1135–1141 1139

123

The type strain, MBRL 179T (=JCM

17576T = KCTC 29124T), was isolated from a lime-

stone quarry at Hundung, Manipur, India. The 16S

rRNA gene sequences of strain MBRL 179T has been

deposited in GenBank under the accession number

JN560155.

Acknowledgments The authors are grateful to Prof.

Tomohiko Tamura (NBRC) for providing the reference type

strains. S Nimaichand wishes to thank the University Grants

Commission (UGC), Government of India, for offering him the

Rajiv Gandhi National Fellowship. K Tamreihao wishes to

thank the Council of Scientific and Industrial Research (CSIR),

Government of India for offering him the CSIR-SRF. Y-G

Zhang was supported by the West Light Foundation of the

Chinese Academy of Sciences. W-J Li was also supported by

‘Hundred Talents Program’ of the Chinese Academy of

Sciences. The authors also wish to thank SAIF, NEHU,

Shillong for providing the SEM facility.

References

Brautaset T, Sekurova ON, Sletta H, Ellingsen TE, Strøm AR,

Valla S, Zotchev SB (2000) Biosynthesis of the polyene

antifungal antibiotic nystatin in Streptomyces noursei

ATCC 11455: analysis of the gene cluster and deduction of

the biosynthetic pathway. Chem Biol 7:395–403

Caffrey P, Lynch S, Flood E, Finnan S, Oliynk M (2001)

Amphotericin biosynthesis in Streptomyces nodosus:

deductions from analysis of polyketide synthase and late

genes. Chem Biol 8:713–723

Collins MD, Pirouz T, Goodfellow M, Minnikin DE (1977)

Distribution of menaquinones in actinomycetes and cory-

nebacteria. J Gen Microbiol 100:221–230

Collins CH, Lyne PM, Grange JM, Falkinham JO (2004)

Microbiological methods, 8th edn. Arnold, London

DeLey J, Caltoir H, Reeynaerts A (1970) The quantitative

measurement of DNA hybridization from renaturation rate.

Eur J Biochem 12:133–142

Diab A, Al-Gounaim MY (1982) Streptomyces spinoverruco-

sus, a new species from the air of Kuwait. Int J Syst Bact

32:327–331

Euzeby JP (2013). List of bacterial names with standing in

nomenclature: a folder available on the internet. (Last full

update: 14 May 2013), http://www.bacterio.cict.fr/

Felsenstein J (1985) Confidence limits on phylogenies: an

approach using the bootstrap. Evolution 39:783–791

Goodfellow M (1986) Genus Rhodococcus Zopf 1891, 28AL. In:

Sneath PHA, Mair NS, Sharpe NE, Holt JG (eds) Bergey’s

manual of systematic bacteriology, vol 2. Williams &

Wilkins, Baltimore, pp 1472–1481

Gordon RE, Barnett DA, Handerhan JE, Pang CHN (1974)

Nocardia coeliaca, Nocardia autotrophica, and the no-

cardin strain. Int J Syst Bacteriol 24:54–63

Hamdali H, Halidi M, Virolle MJ, Qahdouch Y (2008) Rock

phosphate-solubilizing actinomycetes: screening for plant

growth-promoting activities. World J Microbiol Biotech-

nol 24:2565–2575

Kampfer P (2012) Genus I. Streptomyces Waksman and Hen-

rici 1943, 339 emend. Witt and Stackebrandt 1990, 370

emend. Wellington, Stackebrandt, Sanders, Wolstrup and

Jorgensen 1992, 159. In: Goodfellow M, Kampfer P,

Busse H-J et al (eds) Bergey’s manual of systematic

bacteriology, Part B, vol 5, 2nd edn. Springer, New York,

pp 1455–1767

Kelly KL (1964) Inter-Society Color Council—National Bureau

of Standards Color-Name Charts Illustrated with Centroid

Colors. US Government Printing Office, Washington

Khamna S, Yokota A, Lumyong S (2009) Actinomycetes iso-

lated from medicinal plant rhizospheric soils: diversity and

screening of antifungal compounds, indole-3-acetic acid

and siderophore production. World J Microbiol Biotechnol

25:649–655

Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon

YS, Lee JH, Yi H, Won S, Chun J (2012) Introducing

EzTaxon-e: a prokaryotic 16S rRNA Gene sequence

database with phylotypes that represent uncultured species.

Int J Syst Evol Microbiol 62:716–721

Kimura M (1983) The neutral theory of molecular evolution.

Cambridge University Press, Cambridge

Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan

PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lo-

pez R, Thompson JD, Gibson TJ, Higgins DG (2007)

Clustal W and Clustal X version 2.0. Bioinformatics

23:2947–2948

Li WJ, Xu P, Schumann P, Zhang YQ, Pukall R, Xu LH,

Stackebrandt E, Jiang CL (2007) Georgenia ruanii sp.

nov., a novel actinobacterium isolated from forest soil in

Yunnan (China), and emended description of the genus

Georgenia. Int J Syst Evol Microbiol 57:1424–1428

Mesbah M, Premachandran U, Whitman WB (1989) Precise

measurement of the G ? C content of deoxyribonucleic

acid by high-performance liquid chromatography. Int J

Syst Bacteriol 39:159–167

Minnikin DE, O’Donnell AG, Goodfellow M, Alderson G,

Athalye M, Schaal A, Parlett JH (1984) An integrated

procedure for the extraction of bacterial isoprenoid qui-

nones and polar lipids. J Microbiol Methods 2:233–241

Nimaichand S, Zhu WY, Yang LL, Ming H, Nie GX, Tang SK,

Ningthoujam DS, Li WJ (2012) Streptomyces manipuren-

sis sp. nov., a novel actinobacterium isolated from a

limestone deposit site in Manipur, India. Antonie Van

Leeuwenhoek 102:133–139

Saitou N, Nei M (1987) The neighbour-joining method: a new

method for reconstructing phylogenetic trees. Mol Bio

Evol 4:406–425

Sasser M (1990) Identification of bacteria by gas chromatog-

raphy of cellular fatty acids. USFCC Newsl 20:16

Shirling EB, Gottlieb D (1966) Methods for characterization of

Streptomyces species. Int J Syst Bacteriol 16:313–340

Sierra GA (1957) A simple method for the detection of lipolytic

activity of micro-organisms and some observations on the

influence of the contact between cells and fatty substrates.

Antonine van Leeuwenhoek 23:15–22

Staneck JL, Roberts GD (1974) Simplified approached to

identification of aerobic actinomycetes by thin-layer

chromatography. Appl Microbiol 28:226–231

1140 Antonie van Leeuwenhoek (2013) 104:1135–1141

123

Tamaoka J, Katayama-Fujimura Y, Kuraishi H (1983) Analysis

of bacterial menaquinone mixtures by high performance

liquid chromatography. J Appl Bacteriol 54:31–36

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S

(2011) MEGA5: molecular evolutionary genetics analysis

using maximum likelihood, evolutionary distance, and

maximum parsinomy methods. Mol Bio Evol 28:2731–2739

Tang SK, Wang Y, Chen Y, Lou K, Cao LL, Xu LH, Li WJ

(2009) Zhihengliuella alba sp. nov., and emended

description of the genus Zhihengliuella. Int J Syst Evol

Microbiol 59:2025–2031

Wayne LG, Brenner DJ, Colwell RR, Grimont PAD, Kandler O,

Krichevsky MI, Moore LH, Moore WEC, Murray RGE,

Stackebrandt E, Starr MP, Truper HG (1987) International

committee on systematic bacteriology. Report of the ad hoc

committee on reconciliation of approaches to bacterial

systematic. Int J Syst Bacteriol 37:463–464

Williams ST, Davies FL (1967) Use of a scanning electron

microscope for the examination of actinomycetes. J Gen

Microbiol 48:171–177

Yuan WM, Crawford DL (1995) Characterization of Strepto-

myces lydicus WYEC108 as a potential biocontrol agent

against fungal root and seed rots. Appl Environ Microbiol

61:3119–3128

Antonie van Leeuwenhoek (2013) 104:1135–1141 1141

123