micromonospora taraxaci sp. nov., a novel endophytic actinomycete isolated from dandelion root...
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ORIGINAL PAPER
Micromonospora taraxaci sp. nov., a novel endophyticactinomycete isolated from dandelion root (Taraxacummongolicum Hand.-Mazz.)
Junwei Zhao • Lifeng Guo • Hairong He •
Chongxi Liu • Yuejing Zhang • Chuang Li •
Xiangjing Wang • Wensheng Xiang
Received: 25 April 2014 / Accepted: 14 July 2014 / Published online: 1 August 2014
� Springer International Publishing Switzerland 2014
Abstract A novel actinomycete, designated strain
NEAU-P5T, was isolated from dandelion root (Tarax-
acum mongolicum Hand.-Mazz.). Strain NEAU-P5T
showed closest 16S rRNA gene sequence similarity to
Micromonospora chokoriensis 2–19/6T (99.5 %), and
phylogenetically clustered with Micromonospora vio-
lae NEAU-zh8T (99.3 %), M. saelicesensis Lupac 09T
(99.0 %), M. lupini Lupac 14NT (98.8 %), M. zeae
NEAU-gq9T (98.4 %), M. jinlongensis NEAU-
GRX11T (98.3 %) and M. zamorensis CR38T
(97.9 %). Phylogenetic analysis based on the gyrB
gene sequence also indicated that the isolate clustered
with the above type strains except M. violae NEAU-
zh8T. The cell-wall peptidoglycan consisted of meso-
diaminopimelic acid and glycine. The major menaqui-
nones were MK-9(H8), MK-9(H6) and MK-10(H2). The
phospholipid profile contained diphosphatidylglycerol,
phosphatidylethanolamine and phosphatidylinositol.
The major fatty acids were C16:0, iso-C15:0 and C17:0.
Furthermore, some physiological and biochemical
properties and low DNA–DNA relatedness values
enabled the strain to be differentiated from members
of closely related species. Therefore, it is proposed that
strain NEAU-P5T represents a novel species of the
genus Micromonospora, for which the name Micromo-
nospora taraxaci sp. nov. is proposed. The type strain is
NEAU-P5T (=CGMCC 4.7098T = DSM 45885T).
Keywords Micromonospora taraxaci sp. nov. �Polyphasic taxonomy � 16S rRNA gene � gyrB gene
Introduction
Endophytic actinobacteria, which are ubiquitous in
living plants, have aroused more and more attention in
recent years. These microorganisms may be underex-
plored sources of novel species and novel natural
products for exploitation in pharmaceutical and agri-
culture (Qin et al. 2009, 2011). Dandelion is a
Junwei Zhao and Lifeng Guo have contributed equally to this
work.
Electronic supplementary material The online version ofthis article (doi:10.1007/s10482-014-0237-x) contains supple-mentary material, which is available to authorized users.
J. Zhao � L. Guo � H. He � C. Liu � Y. Zhang �C. Li � X. Wang (&) � W. Xiang (&)
Key Laboratory of Agriculture Biological Functional
Gene of Heilongjiang Provincial Education Committee,
Northeast Agricultural University, No. 59 Mucai Street,
Xiangfang District, Harbin 150030, People’s Republic of
China
e-mail: [email protected]
W. Xiang
e-mail: [email protected]
W. Xiang
State Key Laboratory for Biology of Plant Diseases and
Insect Pests, Institute of Plant Protection, Chinese
Academy of Agricultural Sciences, Beijing, People’s
Republic of China
123
Antonie van Leeuwenhoek (2014) 106:667–674
DOI 10.1007/s10482-014-0237-x
traditional Chinese medicine, which was widely used
to treat liver diseases, kidney diseases and spleen
problems (Ling and Zheng 1998). During the inves-
tigation of exploring potential sources of actinomy-
cetes with novel natural products in the root of
dandelion (Taraxacum mongolicum Hand.-Mazz.), a
total of 15 isolates of endophytic actinomycetes were
isolated from dandelion root sample. Out of 15
isolates, Streptomyces was the dominant genus
(n = 9, 60 % of isolates), followed by Micromonos-
pora sp. (n = 5, 33.33 %) and Streptosporangium sp.
(n = 1, 6.67 %). In this study, we performed poly-
phasic taxonomy on strain NEAU-P5T, and proposed
that the novel isolate is a new species of the genus
Micromonospora, for which the name Micromonos-
pora taraxaci sp. nov. is proposed.
Materials and methods
Isolation and maintenance of the organisms
Strain NEAU-P5T was isolated from dandelion root (T.
mongolicum Hand.-Mazz.) collected from Harbin, Hei-
longjiang province, north China (45�450N, 126�410E).
The root sample was processed as described by Wang
et al. (2013) and placed on a plate of humic acid-vitamin
agar (HV) (Hayakawa and Nonomura 1987) supple-
mented with cycloheximide (50 mg L-1) and nalidixic
acid (50 mg L-1). After 2 weeks of aerobic incubation at
28 �C, colonies were transferred and purified on Interna-
tional Streptomyces Project (ISP) medium 3 (Shirling and
Gottlieb 1966) and maintained as glycerol suspensions
(20 %, v/v) at -80 �C.
Antifungal activity bioassay in vitro
Strain NEAU-P5T was point-inoculated at the periph-
ery of potato dextrose agar (PDA: potato 200 g,
dextrose 20 g, agar 20 g, distilled water 1 L), carrot
agar (CA: carrot 200 g; agar 20 g; distilled water 1 L)
and rye sucrose agar (RSA: rye 60 g, sucrose 18 g,
agar 20 g, distilled water 1 L) plates and incubated for
7 days at 28 �C, after which a fresh mycelial PDA, CA
and RSA agar plug of fungi was transferred to the
center of the corresponding plate. After an additional
7 days of incubation at 18 and 28 �C, respectively,
inhibition of hyphal growth of fungal strains was
scored (Mendes et al. 2011). The fungal strains:
Phytophthora sojae (CA medium, 28 �C) was kindly
provided by Soybean Research Institute of Northeast
Agricultural University (Harbin, China); Alternaria
solani (PDA medium, 28 �C), Corynespora cassiicola
(PDA medium, 28 �C), Phytophthora infestans (RSA
medium, 18 �C), Colletotrichum orbiculare (PDA
medium, 28 �C), Rhizoctonia solani (PDA medium,
28 �C), Sclerotinia sclerotiorum (PDA medium,
18 �C) and P. capsici (CA medium, 18 �C) were
kindly provided by the Institute of Vegetables and
Flowers, Chinese Academy of Agricultural Sciences
(Beijing, China).
Morphological, cultural, physiological
characteristics
Morphological characteristics were observed by light
microscopy (Nikon ECLIPSE E200) and scanning
electron microscopy (Hitachi S-3400 N) after cultivation
on ISP 3 medium at 28 �C for 3 weeks. Cultural
characteristics were determined after growth for 3 weeks
at 28 �C on SA1 agar (Trujillo et al. 2005), N-Z amine
agar (Trujillo et al. 2007), Bennett’s agar (Jones 1949),
ISP media 2–7 (Shirling and Gottlieb 1966). ISCC-NBS
colour charts Standard Samples No 2106 (Kelly 1964)
was used to determine the color of colonies and soluble
pigments. The growth temperature range (4, 10, 15, 20,
28, 35, 37, 40, 45 �C) was determined on ISP 3 medium
after culturing for 2 weeks. Tolerance of various pH (3, 4,
5, 6, 7, 8, 9, 10, 11, 12) and NaCl (0, 1, 2, 3, 4 and 5 %,
w/v) were determined in GY medium (Jia et al. 2013) in
shake flasks (250 r.p.m.) at 28 �C for 7 days. Reduction
of nitrate, degradation of gelatin, cellulose and urea,
hydrolysis of starch and aesculin, coagulation of milk and
production of catalase and H2S were examined as
described by Gordon et al. (1974). Utilization of sole
carbon sources was tested on ISP 9 medium (Shirling and
Gottlieb 1966). Utilization of amino acids as nitrogen
sources was tested as described by Williams et al. (1983).
The reference strains M. chokoriensis 2–19(6)T, M.
saelicesensis Lupac 09T, M. lupini Lupac 14NT and M.
zamorensis CR38T were obtained from Japan Collection
of Microorganisms (JCM) and DSMZ (German Collec-
tion of Microorganisms and Cell Cultures), M. violae
NEAU-zh8T (Zhang et al. 2014), M. zeae NEAU-gq9T
(Shen et al. 2014) and M. jinlongensis NEAU-GRX11T
(Gao et al. 2014) were from our laboratory. These strains
were cultured under the same conditions for comparative
analyses.
668 Antonie van Leeuwenhoek (2014) 106:667–674
123
Chemotaxonomic characterization
Biomass for chemical studies was obtained by growing
the strain in GY medium in shake flasks (250 r.p.m.) at
28 �C for 7 days. Cells were harvested by centrifuga-
tion, then washed with distilled water and freeze-dried.
Phospholipids in cells were extracted and identified
using the method of Minnikin et al. (1984). Whole-cell
sugar composition was analyzed according to the
method of Lechevalier and Lechevalier (1980). The
isomer of diaminopimelic acid (DAP) in the whole cell
hydrolysates was derivatised according to McKerrow
et al. (2000) and analyzed by a HPLC method.
Menaquinones were extracted from freeze-dried bio-
mass and purified according to Collins (1985) then
analyzed by a HPLC–UV method as described previ-
ously (Gao et al. 2014). Mycolic acids were checked by
the acid methanolysis method as described previously
(Minnikin et al. 1980). Biomass for fatty acids analysis
was obtained by growing strain NEAU-P5T in GY
medium at 28 �C for 7 days and detected by GC–MS
using the method of Gao et al. (2014).
DNA preparation, amplification and determination
of 16S rRNA gene sequence
Genomic DNA of strain NEAU-P5T was extracted as
described previously by Lee et al. (2003) and PCR
amplification of 16S rRNA gene was carried out using
the method of Loqman et al. (2009). The PCR product
was purified and cloned into the pMD19-T vector
(Takara) and sequenced using an Applied Biosystems
DNA sequencer (model 3730XL) and software provide
by the manufacturer. Almost full-length 16S rRNA
gene sequence (1,509 nt) was obtained and aligned
with multiple sequences obtained from the GenBank/
EMBL/DDBJ databases using Clustal X 1.83. The
alignment was manually verified and adjusted prior to
the construction of phylogenetic trees. Phylogenetic
trees were generated with the neighbour-joining (Sa-
itou and Nei 1987) and maximum-likelihood (Felsen-
stein 1981) algorithms using Molecular Evolutionary
Genetics Analysis (MEGA) software version 5.05
(Tamura et al. 2011). The stability of the clades in the
trees was appraised by bootstrap analysis with 1,000
replicates (Felsenstein 1985). A distance matrix was
generated using the Kimura’s two-parameter model
(Kimura 1980). All positions containing gaps and
missing data were eliminated from the dataset
(complete deletion option). 16S rRNA gene sequence
similarities between strains were calculated on the
basis of pairwise alignment using the EzTaxon-e server
(Kim et al. 2012). PCR amplification of the gyrB gene
was carried out using primers GYF1 and GYR3B
(Garcia et al. 2010). Sequencing and phylogenetic
analysis was performed as described above. Signature
nucleotide pattern in the 16S rRNA gene of the novel
strain was determined after manual verification of the
CLUSTAL_X sequence alignment (Zhi et al. 2009).
Nucleotide positions were numbered according to the
corresponding position in the 16S rRNA gene sequence
of Escherichia coli (Brosius et al. 1978).
DNA base composition and DNA–DNA
hybridization
The G?C content of the genomic DNA was deter-
mined by the thermal denaturation method as described
by Mandel and Marmur (1968), with E. coli JM109 as
the reference strain.
DNA–DNA hybridization was carried out between
strain NEAU-P5T and M. chokoriensis 2–19/6T, M.
violae NEAU-zh8T, M. saelicesensis Lupac 09T, M.
lupini Lupac 14NT, M. zeae NEAU-gq9T, M. jinlong-
ensis NEAU-GRX11T and M. zamorensis CR38T in a
model Cary 100 Bio UV/VIS-spectrophotometer
equipped with a Peltier-thermostatted 6 9 6 multicell
changer and a temperature controller with in situ
temperature probe (Varian) as described previously
(De Ley et al. 1970; Huss et al. 1983). The DNA
concentration was adjusted spectrophotometrically at
260 nm to around 1 using 0.1 9 saline sodium citrate
(SSC). The renaturation rates of sheared DNA were
determined at 70 �C.
Results and discussion
Morphological observation of a 21-day-old culture of
strain NEAU-P5T grown on ISP3 agar revealed it had
the typical characteristics of genus Micromonospora.
Substrate mycelium was well-developed without frag-
mentation. Spores (0.6 9 0.8 lm) were borne singly
on the substrate mycelia and the spore surface was
smooth (Suppl. Fig. S1). Good growth was observed on
N-Z amine, ISP2, ISP3 and ISP4 agar; moderate
growth was observed on SA1, Bennett’s, ISP5 and
Antonie van Leeuwenhoek (2014) 106:667–674 669
123
ISP7 agar and poor growth was observed on ISP6 agar.
The colour of colonies was dark orange yellow on N-Z
amine agar, strong yellowish brown on Bennett’s agar,
strong orange yellow on SA1 agar, deep orange yellow
on ISP2 agar, moderate orange yellowish brown on
ISP3 agar, brilliant orange yellow on ISP4 agar, deep
yellow on ISP5 agar, moderate yellow on ISP6 agar and
light yellow on ISP7 agar. No aerial mycelium was
observed on any of the tested media. Grayish purple
and light grayish purplish red soluble pigments were
observed on ISP3 and ISP5 agar, respectively. Melanin
was not observed on ISP6 and ISP7 agar. Growth of
strain NEAU-P5T occurred in the pH range 6–12 and
0–2 % NaCl (w/v), with optimum growth at pH 7.0 and
0 % NaCl (w/v). The temperature range for growth was
10–37 �C, with the optimum temperature being 28 �C.
Detailed physiological and biochemical properties are
presented in the species description. No antifungal
activity of strain NEAU-P5T was observed in this
study. Micromonospora species are also able to
produce antitumor compounds, for example, the
endophytic M. lupini Lupac 08T, which is closely
related to strain NEAU-P5T, produces two novel
antitumoral compounds lupinadicins A and B (Igarashi
et al. 2007). Therefore, the antitumor activity of
NEAU-P5T will be assayed in the following research.
Cells of strain NEAU-P5T were observed to contain
meso-diaminopimelic acid and glycine as diagnostic
amino acids. Whole-cell hydrolysates were found to
contain rhamnose, xylose, glucose and galactose. The
major menaquinones detected were MK-9(H8)
(50.5 %), MK-9(H6) (10.6 %), MK-10(H2) (10.1 %),
MK-8(H8) (6 %), MK-9(H10) (5.9 %), MK-10(H8)
(5.7 %), MK-10(H4) (5.6 %), MK-9(H4) (3.6 %) and
MK-9(H2) (1.9 %). The phopholipids was found to
consist of diphosphatidylglycerol, phosphatidyletha-
nolamine, phosphatidylinositol and an unknown phop-
holipid (Suppl. Fig. S2). The cellular fatty acid profile
was determined to be composed of C16:0 (27.2 %), iso-
C15:0 (23.3 %), C17:0 (13.0 %), C15:0 (6.6 %), C17:1
x7c (6.2 %), anteiso-C17:0 (5.7 %), C18:0 (5.3 %),
C17:0 cyclo (4.6 %), C18:1 x9c (2.9 %), C17:0
10-methyl (1.9 %), iso-C14:0 (1.3 %), C16:1 x7c
(1.2 %) and C18:0 10-methyl (0.7 %). Mycolic acids
were not detected.
The almost complete 16S rRNA gene sequence
(1,509 nt) of strain NEAU-P5T was determined and
deposited as KC439463 in the GenBank/EMBL/DDBJ
databases. Comparative 16S rRNA gene sequence
analysis showed that strain NEAU-P5T was phyloge-
netically related to members of the genus Micromo-
nospora. The position of strain NEAU-P5T in the
family Micromonosporaceae was also supported by
the presence of a complete set of family-specific
signature nucleotides (Zhi et al., 2009) in its 16S rRNA
gene sequence (Suppl. Table S1). Phylogenetic ana-
lysis based on the 16S rRNA gene sequence indicated
that strain NEAU-P5T clustered with M. chokoriensis
2–19/6T (99.5 %), M. violae NEAU-zh8T (99.3 %), M.
saelicesensis Lupac 09T (99.0 %), M. lupini Lupac
14NT (98.8 %), M. zeae NEAU-gq9T (98.4 %), M.
jinlongensis NEAU-GRX11T (98.3 %) and M. zamor-
ensis CR38T (97.9 %), which was supported by a
bootstrap value of 63 % in the neighbour-joining tree
(Fig. 1) and also recovered with the maximum-likeli-
hood algorithm. Partial sequence of gyrB gene was
obtained (1,162 nt) and deposited as KJ608999 in the
GenBank/EMBL/DDBJ databases. The similarities of
gyrB nucleotide sequence between strain NEAU-P5T
and M. chokoriensis 2–19/6T, M. violae NEAU-zh8T,
M. saelicesensis Lupac 09T, M. lupini Lupac 14NT, M.
zeae NEAU-gq9T, M. jinlongensis NEAU-GRX11T
and M. zamorensis CR38T were 98.0, 92.9, 94.9, 95.2,
94.7, 94.8, and 94.7 %, respectively. Phylogenetic
analysis of gyrB nucleotide sequence supported that
strain NEAU-P5T was placed in the genus Micromo-
nospora, near to M. chokoriensis 2–19/6T, M. saelic-
esensis Lupac 09T, M. lupini Lupac 14NT, M. zeae
NEAU-gq9T, M. jinlongensis NEAU-GRX11T and M.
zamorensis CR38T by a 66 % bootstrap value but far
away from M. violae NEAU-zh8T (Suppl. Fig. S3).
Other studies have also reported similar results that the
topology of phylogenetic trees based on gyrB gene
sequences differed from those based on 16S rRNA
gene sequence data (Wang et al. 2011; Carro et al.
2012; Li et al. 2014). To establish the precise
taxonomic position of strain NEAU-P5T, DNA–DNA
hybridizations performed between the novel isolate
and M. chokoriensis 2–19/6T, M. violae NEAU-zh8T,
M. saelicesensis Lupac 09T, M. lupini Lupac 14NT, M.
zeae NEAU-gq9T, M. jinlongensis NEAU-GRX11T
and M. zamorensis CR38T; the levels of DNA–DNA
relatedness between them were 53.8 ± 3.9,
57.1 ± 2.5, 33.8 ± 1.1, 54.1 ± 2.7, 41.9 ± 1.2,
34.6 ± 1.9 and 38.5 ± 2.1 %, respectively. These
values were below the threshold value of 70 %
recommended by Wayne et al. (1987) for assigning
strains to the same genomic species.
670 Antonie van Leeuwenhoek (2014) 106:667–674
123
Besides the genotypic evidence above, the strain
could also be distinguished from the seven related
Micromonospora type strains by morphological,
physiological and biochemical characteristics as sum-
marized in Table 1, such as the clearly different
colony colours on ISP2, ISP3 and ISP4 media after
being incubated at 28 �C for 3 weeks (Suppl. Fig. S4)
and the differences in production of urease, hydrolysis
of starch, decomposition of cellulose and patterns of
carbon and nitrogen utilization (Table 1). Therefore,
strain NEAU-P5T represents a novel species of the
genus Micromonospora, for which the name Micro-
monospora taraxaci sp. nov. is proposed. It is quite
interesting that several Micromonospora species iso-
lated from different endophytic root origins are
clustering together after 16S rRNA and gyrB gene
sequencing and worthy of further research.
Description of Micromonospora taraxaci sp.nov
Micromonospora taraxaci (ta.ra.xa’ci. L. n. Taraxa-
cum, name of a plant, and also a botanical generic
name (Taraxacum); L. gen. n. taraxaci of Taraxacum,
referring to the isolation of the organism from T.
mongolicum Hand.-Mazz.).
Aerobic, Gram-staining positive actinomycete that
forms extensively branched, non-fragmenting substrate
mycelium but no aerial hypha. Good growth is observed
on N-Z amine, ISP2, ISP3 and ISP4 agar; moderate
growth is observed on SA1, Bennett’s, ISP5 and ISP7
agar; poor growth is observed on ISP6 agar. Colonies are
orange yellow colour-series. Grayish purple and light
grayish purplish red soluble pigments are observed on
ISP3 and ISP5 agar, respectively. Spores are single, oval
with smooth surface. Growth occurs at pH values
Fig. 1 Neighbour-joining
tree based on 16S rRNA gene
sequence (1,354 nt) showing
relationship between strain
NEAU-P5T and members of
the genus Micromonospora.
The out-group used was
Catellatospora citrea subsp.
citrea DSM 44097T. Only
bootstrap values above 50 %
(percentages of 1,000
replications) are indicated.
Asterisks indicate branches
also recovered in the
maximum-likelihood tree;
Bar 0.005 nucleotide
substitutions per site
Antonie van Leeuwenhoek (2014) 106:667–674 671
123
between 6 and 12, the optimum being pH 7.0. Tolerates
up to 2 % NaCl and grows at temperatures between 10
and 37 �C, with an optimum temperature of 28 �C.
Positive for liquefaction of gelatin, hydrolysis of aescu-
lin, starch, cellulose and Tween 40, production of
catalase and urease and negative for reduction of nitrate,
coagulation of milk, hydrolysis of Tween 20 and 80 and
production of H2S. L-arabinose, D-galactose, D-glucose,
lactose, D-maltose, D-mannose, D-raffinose, D-mannitol,
D-sorbitol D-ribose, D-sucrose and D-xylose are utilized as
sole carbon sources but D-fructose, inositol and L-
rhamnose are not. L-alanine, L-arginine, L-asparagine,
L-aspartic acid, L-threonine, creatine, L-glutamic acid and
L-serine are utilized as sole nitrogen sources but L-
glutamine, glycine and L-tyrosine are not. No inhibition
was observed against A. solani, C. cassiicola, P.
infestans, C. orbiculare, P. sojae, R. solani, S. sclerotio-
rum and P. capsici. Cell wall contains meso-diamino-
pimelic acid and glycine, and the characteristic whole-
cell sugars are rhamnose, xylose, glucose and galactose.
The phopholipids include diphosphatidylglycerol,
phosphatidylethanolamine, phosphatidylinositol and
an unknown phopholipid. The major menaquinones
are MK-9(H8), MK-9(H6) and MK-10(H2). The major
cellular fatty acids are C16:0, iso-C15:0 and C17:0.
Mycolic acids are absent. The G?C content of the
DNA of the type strain is 68.3 ± 0.25 mol %.
The type strain, NEAU-P5T (=CGMCC
4.7098T = DSM 45885T), was isolated from a root
of dandelion (T. mongolicum Hand.-Mazz.) collected
from Harbin, Northeast China. The GenBank/EMBL/
DDBJ accession numbers for the 16S rRNA gene and
gyrB gene sequences of strain NEAU-P5T are
KC439463 and KJ608999, respectively.
Acknowledgments This work was supported in part by grants
from the National Outstanding Youth Foundation (No.
31225024), the National Key Project for Basic Research (No.
2010CB126102), the National Key Technology R&D Program
(No. 2012BAD19B06), the Program for New Century Excellent
Talents in University (NCET-11-0953), the National Natural
Science Foundation of China (No.31372006, 31171913 and
31071750), the Outstanding Youth Foundation of Heilongjiang
Province (JC201201), the Chang Jiang Scholar Candidates
Program for Provincial Universities in Heilongjiang (CSCP),
the Sicence and Technology Research Project of Heilongjiang
Table 1 Differential characteristics of strain NEAU-P5T and the closely related species
Characteristics 1 2 3 4 5 6 7 8
Spore ornamentation Smooth Rough to nodulara Smooth Smoothb Smoothb Smooth Smooth Smoothc
Maximum NaCl tolerance (%, w/v) 2 2 2 3 3 3 3 3
Cellulose decomposition ? ? - ? ? - - -
Production of urease ? ? ? ? ? - - ?
Hydrolysis of starch ? - - - - - ? -
Use as sole carbon source:
D-maltose ? ? ? - ? ? ? -
D-mannose ? ? ? ? ? ? - -
D-sorbitol ? - - - - - ? -
L-rhamnose - ? - ? ? ? ? ?
D-mannitol ? ? - - - ? ? -
Use as sole nitrogen source:
Creatine ? ? - ? ? ? - ?
L-threonine ? - - ? ? ? ? ?
L-glutamic acid ? - ? ? - - ? -
L-glutamine - ? ? ? ? ? ? ?
Strains: 1, NEAU-P5T; 2, M. chokoriensis 2–19(6)T; 3, NEAU-zh8T; 4, M. saelicesensis Lupac 09T; 5, M. lupini Lupac 14NT; 6, M.
zeae NEAU-gq9T; 7, M. jinlongensis NEAU-GRX11T; 8, M. zamorensis CR38T. (?) positive, (-) negative. Data are from this study
except where markeda Data from Ara and Kudo (2007)b Data from Trujillo et al. (2007)c Data from Carro et al. (2012)
672 Antonie van Leeuwenhoek (2014) 106:667–674
123
Provincial Educational Commission (No. 12541001) and the
Youth Science Foundation of Heilongjiang Province (No.
QC2014C013).
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