Antonie van Leeuwenhoek

, Volume 103, Issue 2, pp 399–408

Wangella harbinensis gen. nov., sp. nov., a new member of the family Micromonosporaceae

Authors

  • Feiyu Jia
    • School of Life ScienceNortheast Agricultural University
  • Chongxi Liu
    • School of Life ScienceNortheast Agricultural University
  • Xiangjing Wang
    • School of Life ScienceNortheast Agricultural University
  • Junwei Zhao
    • School of Life ScienceNortheast Agricultural University
  • Qianfeng Liu
    • School of Life ScienceNortheast Agricultural University
  • Ji Zhang
    • School of Life ScienceNortheast Agricultural University
  • Ruixia Gao
    • School of Life ScienceNortheast Agricultural University
    • School of Life ScienceNortheast Agricultural University
Original Paper

DOI: 10.1007/s10482-012-9820-1

Cite this article as:
Jia, F., Liu, C., Wang, X. et al. Antonie van Leeuwenhoek (2013) 103: 399. doi:10.1007/s10482-012-9820-1

Abstract

A novel endophytic actinomycete, designated strain NEAU-J3T, was isolated from soybean root (Glycine max (L.) Merr) and characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain NEAU-J3T fell within the family Micromonosporaceae. The strain was observed to form an extensively branched substrate mycelium, which carried non-motile oval spores with a smooth surface. The cell walls of strain NEAU-J3T were determined to contain meso-diaminopimelic acid and galactose, ribose and glucose were detected as whole-cell sugars. The major menaquinones were determined to be MK-9(H4) and MK-9(H6). The phospholipids detected were phosphatidylcholine and phosphatidylethanolamine. The major cellular fatty acids were determined to be C16:0, C18:1 ω9c, C18:0, C17:0, C17:1 ω7c, anteiso-C17:0, C16:1 ω7c and C15:0. The DNA G + C content was 62.5 mol%. On the basis of the morphological and chemotaxonomic characteristics, phylogenetic analysis and characteristic patterns of 16S rRNA gene signature nucleotides, strain NEAU-J3T is considered to represent a novel species of a new genus within the family Micromonosporaceae, for which the name Wangella harbinensis gen. nov., sp. nov. is proposed. The type strain of Wangella harbinensis is strain NEAU-J3T (=CGMCC 4.7039T = DSM 45747T).

Keywords

Wangella harbinensis gen. nov. Sp. nov.Polyphasic taxonomy16S rRNA gene

Introduction

The family Micromonosporaceae was first described by Krasil’nikov (1938) and its description has been subsequently emended by Goodfellow et al. (1990), Koch et al. (1996), Stackebrandt et al. (1997) and Zhi et al. (2009) on the basis of chemotaxonomic data and 16S rRNA gene sequence analysis. At the time of writing, the family Micromonosporaceae comprises 27 genera. During a study on the ecological diversity of actinomycetes from soybean root, an aerobic actinomycete strain, NEAU-J3T, was isolated from soybean root collected from Harbin, Heilongjiang Province, north China. In this study, the taxonomic status of this strain is reported based on phylogenetic, chemotaxonomic and phenotypic evidence. It is proposed that strain NEAU-J3T is placed in a new genus and species of the family Micromonosporaceae, designated Wangella harbinensis.

Materials and methods

Isolation and maintenance of the organism

Strain NEAU-J3T was isolated from soybean root (Glycine max (L.) Merr) collected from Harbin, Heilongjiang province, north China (45°45′ N, 126°41′ E). The plant was tagged outdoors and stored in a clean plastic bag until used (approx. 24 h). The root sample was air dried for 24 h at room temperature and then washed in water with an ultrasonic step (160 W, 15 min) to remove the surface soils and adherent epiphytes completely. After drying, the sample was cut into pieces of 5–10 mm in length and then subjected to a seven-step surface sterilization procedure: a 60-s wash in sterile tap-water containing cycloheximide (100 mg l−1) and nalidixic acid (20 mg l−1), followed by a wash in sterile water, a 5-min wash in 5 % (v/v) NaOCl, a 10-min wash in 2.5 % (w/v) Na2S2O3, a 5-min wash in 75 % (v/v) ethanol, a wash in sterile water and a final rinse in 10 % (w/v) NaHCO3 for 10 min. The rinsed root sample was transferred temporarily onto sterile filter paper to eliminate excess moisture and then placed on a plate of humic acid-vitamin agar (HV) (Hayakawa and Nonomura 1987) supplemented with cycloheximide (50 mg l−1) and nalidixic acid (20 mg l−1). After 21 days of aerobic incubation at 28 °C, colonies were transferred and purified on oatmeal agar (International Streptomyces Project (ISP) medium 3) and incubated at 28 °C for 2–3 weeks.

Morphological, cultural and physiological characteristics

Morphological properties were observed by light microscopy (Nikon ECLIPSE E200) and scanning electron microscopy (Hitachi S-3400 N) using cultures grown on ISP3 agar at 28 °C for 21 days. Cultural characteristics of strain NEAU-J3T were recorded on ISP media 2–7 (Shirling and Gottlieb 1966), glucose-asparagine agar (glucose 1 %, asparagine 0.05 %, K2HPO4·3H2O 0.05 %, agar 1.5 %, w/v, pH 7.2–7.4), oatmeal-nitrate agar (JCM medium 52), Hickey–Tresner agar (yeast extract 0.1 %, beef extract 0.1 %, NZamine A 0.2 %, dextrin 1 %, CoCl2·6H2O 0.002 %, agar 2 %, w/v, pH 7.2–7.4), tap-water agar (Gordon et al. 1974), sucrose-nitrate agar (Waksman medium 1), yeast extract-starch agar (JCM medium 61), glucose-yeast extract agar (Cross et al. 1968), Bennett’s agar (Jones 1949) and HV agar (Hayakawa and Nonomura 1987). The Color Harmony Manual (Jacobson et al. 1958) was used to determine the names and designations of colony colours. The temperature range for growth was determined on ISP3 medium after incubation for 2 weeks. The pH range and NaCl tolerance for growth were determined on GY medium (glucose 1 %, yeast extract 1 %, K2HPO4·3H2O 0.05 %, MgSO4·7H2O 0.05 %, w/v, pH 7.2) at 28 °C for 14 days on a rotary shaker. Decomposition of cellulose, hydrolysis of starch, reduction of nitrate, liquefaction of gelatin and production of H2S were examined as described previously (Gordon et al. 1974). Utilization of carbohydrates as sole carbon sources was tested using ISP medium 9 as basal medium according to the method of Shirling and Gottlieb (1966). The utilization of amino acids as nitrogen sources was tested as described by Williams et al. (1983).

Chemotaxonomic characterization

Biomass for chemical studies was prepared by growing the strain in tryptic soy broth (TSB: tryptone, 15 g; soya peptone, 5 g; NaCl, 0.5 g; distilled water, 1 l; pH 7.0–7.4) in Erlenmeyer flasks for 14 days at 28 °C. Cells were harvested by centrifugation, washed with distilled water and freeze-dried. The isomer of diaminopimelic acid (DAP) in the whole-cell hydrolysates was analysed by an HPLC method using Agilent TC–C18 Column (250 × 4.6 mm i.d. 5 μm) with a mobile phase consisting of acetonitrile: 0.05 mol l−1 phosphate buffer pH 7.2 = 15:85 at a flow rate of 0.5 ml min−1. The peak detection used an Agilent G1321A fluorescence detector with a 365 nm excitation and 455 nm longpass emission filters (McKerrow et al. 2000). The N-acyl group of muramic acid in peptidoglycan was determined by the method of Uchida et al. (1999). The whole-cell sugars were analyzed according to the procedures developed by Lechevalier and Lechevalier (1980). Phospholipids in cells were extracted and identified by using the method of Minnikin et al. (1984). Menaquinones were extracted from freeze-dried biomass and purified according to Collins (1985). Extracts were analyzed by HPLC–UV method using Agilent Extend-C18 Column (150 × 4.6 mm, i.d. 5 μm), typically at 270 nm. The mobile phase was acetonitrile-propyl alcohol (60:40, v/v) and the flow rate was set to 1.0 ml min−1 and the run time was 60 min. The injection volume was 20 μl, and the chromatographic column was controlled at 40 °C (Wu et al. 1989). Cellular fatty acids were analysed by GC–MS using the method of Xiang et al. (2011).

DNA G + C content, 16S rRNA gene sequencing and determination of the signature nucleotide positions

Genomic DNA of strain NEAU-J3T was extracted as described previously by Lee et al. (2003). The DNA G + C content of the genomic DNA was determined by the thermal denaturation method as described by Mandel and Marmur (1968), and Escherichia coli JM109 was used as the reference strain. PCR amplification of 16S rRNA gene was carried out according to the procedures described by Loqman et al. (2009). The 16S rRNA gene sequence of strain NEAU-J3T was aligned with multiple sequences obtained from the GenBank/EMBL/DDBJ databases using Clustal X 1.83 software. The alignment was manually verified and adjusted prior to the construction of phylogenetic trees. Phylogenetic trees were generated with the neighbour-joining (Saitou and Nei 1987), maximum-parsimony (Fitch 1971) and maximum-likelihood (Felsenstein 1981) algorithms using Mega 5.05 (Tamura et al. 2011). A distance matrix was generated using Kimura’s two-parameter model (Kimura 1980). All positions containing gaps and missing data were eliminated from the dataset (complete deletion option). Streptomyces ambofaciens ATCC 23877T was used as an outgroup. Confidence values for branches of the phylogenetic trees were determined using bootstrap analyses based on 1,000 resamplings (Felsenstein 1985). Nucleotides and nucleotide pairs in the 16S rRNA gene of the novel strain and closely related members of the family Micromonosporaceae were determined after manual verification of the CLUSTAL_X sequence alignment. Nucleotide positions were numbered according to the corresponding position in the 16S rRNA gene sequence of E. coli (Brosius et al. 1978).

Results and discussion

The almost-complete 16S rRNA gene sequence (1,514 nt; GenBank/EMBL/DDBJ accession number JQ750973) of strain NEAU-J3T showed a close relationship with members of family Micromonosporaceae. 16S rRNA gene sequence comparisons revealed that strain NEAU-J3T was most closely related to Couchioplanes caeruleus subsp. azureus IFO 13993T (98.15 %), Krasilnikovia cinnamomea 3-54(41)T (97.95 %), Pseudosporangium ferrugineum 3-44-a-(19)T (97.89 %), Actinoplanes friuliensis DSM 7358T (97.88 %) and Actinoplanes digitatis IFO 12512T (97.63 %). A Neighbour-Joining phylogenetic tree (Fig. 1) indicated that the new isolate belonged to the family Micromonosporaceae and was closely affiliated with the members of the genera Krasilnikovia, Couchioplanes, Pseudosporangium and Actinoplanes, an association that was supported by maximum-likelihood (Supplementary Fig. S1) and maximum-parsimony (Supplementary Fig. S2) algorithms employed and by a 83 % bootstrap value in the neighbour-joining analysis.
https://static-content.springer.com/image/art%3A10.1007%2Fs10482-012-9820-1/MediaObjects/10482_2012_9820_Fig1_HTML.gif
Fig. 1

Neighbour-joining phylogenetic tree derived from nearly complete 16S rRNA gene sequences (1,514 nt) showing the relationship between strain NEAU-J3T and type species of recognized genera of the family Micromonosporaceae. S. ambofaciens ATCC 23877T was used as an outgroup. Stars indicate branches that were also found using maximum parsimony and maximum-likelihood methods. Numbers at branchpoints indicate bootstrap percentages (based on 1,000 replicates); only values >30 % are indicated. Scale bar 0.01 substitutions per nucleotide position

The membership of strain NEAU-J3T in the family Micromonosporaceae was also supported by the presence of an almost complete set of family-specific signature nucleotides (Zhi et al. 2009) in its 16S rRNA gene sequence. However, when the signature nucleotide positions of strain NEAU-J3T were compared with those of its closest relatives, there were several nucleotide pair differences from the genera Couchioplanes, Krasilnikovia, Pseudosporangium, and Actinoplanes (Table 1).
Table 1

Comparison of 16S rRNA gene signature nucleotide positions of strain NEAU-J3T and closely related genera (Nucleotide positions of bases or base pairs are given according to E. coli numbering (Brosius et al. 1978))

Site

NEAU-J3T

Pseudosporangium

Actinoplanes

Couchioplanes

Krasilnikovia

129

C

C

U

C

U

139:224

G–U

G–U

A–U

G–U

A–U

140:223

G–C

G–C

G–U

G–C

U–G

144:178

U–G

U–G

U–A

U–G

U–G

153:168

C–G

C–G

C–G

G/C–G

C–G

154:167

C–G

C–G

C–G

G/C–G

C–G

155:166

C–G

C–G

C–G

G/C–G

C–G

182

U

U

G

G

G

186:190

G–A

C–C

C–A

U–C

U–G

203

U

G

G

G

G

222

U

U

U

U

C

232

G

G

A

G

G

262

G

G

A

G

G

445:489

G–C

G–C

G–C

G–C

C–G

446:488

G–C

G–C

G–C

G–C

C–G

456

C

U

C

U

A

457

A

G

A

A

G

570

C

G

G

G

G

602:636

C–G

C–G

A–U

C–G

C–G

614:626

G–C

G–C

G–C

G–G/C

G–C

615:625

C–G

C–G

G–C

C–G

C–G

616:624

G–U

G–C

G–U

G–U

G–C

631

C

C

U

U/C

C

832:854

G–U

G–U

G–C

G–U

G–U

837:849

G–C

G–C

G–U

G–C

G–C

1001:1039

C–G

C–G

G–C

C–G

C–G

1006

A

A

U

A

A

1011:1018

U–A

C–G

C–G

C–G

U–A

1119:1154

U–A

U–A

U–A

U–A

U–A

1121:1152

G

C

G

G

C

1133:1141

G

C

G

G

C

1256

U

U

C

U

U

1263:1272

G–U

G–U

A–U

A–U

G–U

1451

C

G

C

G

G

1452

A

G

A

G

G

1469

G

U

A

U

U

Morphological observation of a 21-day-old culture of strain NEAU-J3T grown on ISP3 agar revealed it formed extensively branched, non-fragmenting, yellow- to rusty-coloured substrate mycelium. Aerial hyphae and sporangia were not present. Spores (1.60 × 0.80 μm) were observed to be oval and non-motile with a smooth surface and borne singly on the substrate mycelium (Fig. 2). Strain NEAU-J3T was observed to grow well on oatmeal-nitrate, tap-water, yeast extract-starch, glucose-yeast extract, ISP2, ISP3, ISP4 and ISP7 agar; moderately on glucose-asparagine, Bennett’s and HV agar; poorly on sucrose-nitrate and ISP6 agar; no growth was observed on Hickey–Tresner agar and ISP5 medium (Supplementary Table S1). No soluble pigments were found to be produced on any of the media tested. Growth of strain NEAU-J3T was determined to occur in the pH range 4–10 and 0–3 % NaCl (w/v), with optimum growth at pH 7.0 and 1 % NaCl (w/v). The temperature range for growth was determined to be 18–39 °C, with the optimum temperature being 28 °C. Detailed physiological and biochemical properties are presented in the species description. The cell morphology of strain NEAU-J3T is clearly different from members of the genera Couchioplanes, Krasilnikovia, Pseudosporangium and Actinoplanes, which form globose pseudosporangia on the substrate mycelium. Strain NEAU-J3T is also distinguished from the genera Couchioplanes and Actinoplanes by the absence of motile spores.
https://static-content.springer.com/image/art%3A10.1007%2Fs10482-012-9820-1/MediaObjects/10482_2012_9820_Fig2_HTML.jpg
Fig. 2

Scanning electron micrograph of substrate mycelium of strain NEAU-J3T grown on ISP3 agar for 21 days at 28 °C. Scale bar 1 μm

Cells of strain NEAU-J3T were determined to contain meso-diaminopimelic acid as the diamino acid. Whole-cell hydrolysates were found to contain galactose, ribose and glucose (Supplementary Fig. S3). The acyl type of the cell wall polysaccharides was determined to be glycolyl. The predominant menaquinones were found to be MK-9 (H4) (55 %) and MK-9 (H6) (36 %), with MK-10 (H4) (8 %) and MK-9 (H8) (1 %) as minor components. The phospholipids detected were phosphatidylcholine and phosphatidylethanolamine (Supplementary Fig. S4). The major cellular fatty acids were determined to be C16:0 (40.3 %), C18:1 ω9c (11.7 %), C18:0 (9.8 %), C17:0 (8.3 %), C17:1 ω7c (8.3 %), anteiso-C17:0 (8.3 %), C16:1 ω7c (6.2 %) and C15:0 (6.2 %) (Supplementary Table S2; Fig. S5). Strain NEAU-J3T can be readily distinguished from the most closely related genera in the family Micromonosporaceae by the phospholipid type. Members of the genera Couchioplanes, Krasilnikovia, Pseudosporangium and Actinoplanes correspond to phospholipid type PII, in contrast to strain NEAU-J3T, which belongs to phospholipid type PIII (Lechevalier et al. 1981). The fatty acid pattern also differentiates strain NEAU-J3T from the genera Couchioplanes, Krasilnikovia, Pseudosporangium and Actinoplanes, as these genera have a 2d or 2c fatty acid type, in contrast to the novel isolate, which is of type 1a (Kroppenstedt 1985). Furthermore, the major menaquinones, the whole-cell sugars and the G + C content of the DNA of strain NEAU-J3T differ from those of the most closely related genera in the family Micromonosporaceae (Table 2).
Table 2

Characteristics of strain NEAU-J3T (Wangella gen. nov.) and genera in the family Micromonosporaceae

Genus

Diamino acid (s)

Diagnostic sugars

Fatty-acid type

Major menaquinone (s)

Phospholipid type

Wangella

m-DAP

Rib, Glc, Gal

1a

9 (H4, 6)

III

Pseudosporangium

m- and 3-OH-DAP

Ara, Gal, Glc, Man, Xyl, Rib

2d

9 (H6)

II

Couchioplanes

l-Lys

Ara, Gal, Xyl

2c

9 (H4)

II

Actinoplanes

m-DAP

Ara, Xyl

2d

9 (H4), 10 (H4)

II

Krasilnikovia

m-DAP

Gal, Ara, Xyl, Man, Rib

2d

9 (H6, 4, 8)

II

Actinocatenispora

m-DAP

Ara, Gal, Xyl

3b

9 (H4, 6)

II

Allocatelliglobosispora

3-OH-DAP

Glc, Rha, Rib, Xyl, Ara, Gal, Man

3b

10 (H4), 9 (H4), 10 (H6)

II

Asanoa

m-DAP

Ara, Gal, Xyl

2d

10 (H6, 8)

II

Catellatospora

m-DAP3O

Ara, Gal, Xyl or only Xyl

3b

9 (H4, 6), 10 (H4)

II

Catelliglobosispora

m- and 3-OH-DAP

Rha, Man, Xyl, Gal, Glc

3b

10 (H4)

II

Catenuloplanes

l-Lys

Xyl

2c

9 (H8), 10 (H8)

III

Dactylosporangium

m-DAP

Ara, Xyl

3b

9 (H4, 6, 8)

II

Hamadaea

m-DAP

Xyl, Gal, Man, Rib, Ara, Rha

3b

9 (H4)

II

Jishengella

m-DAP

Xyl, Man, Ara, Rib, Glc

3a

9 (H4, 6, 8), 10 (H4)

II

Longispora

m-DAP

Ara, Gal, Xyl

2d

10 (H4, 6)

II

Luedemannella

m-DAP

Gal, Man, Rha, Rib, Xyl, Ara

2d

9 (H6, 4, 2, 8)

II

Micromonospora

m-DAP

Ara, Xyl

3b

10 (H4, 6), 9 (H4, 6)

II

Phytohabitans

m-DAP, l-Lys

Gal, Glc, Man, Rib, Xyl

2d

9 (H6), 10 (H4, 6)

II

Phytomonospora

m-DAP

Gal, Glc, Rib, Man

2d

8 (H2), 9 (H2), 10 (H2, 4, 6)

PIII

Pilimelia

m-DAP

Ara, Xyl

2d

9 (H2, 4)

II

Planosporangium

m-DAP

Ara, Gal, Xyl

2d

10 (H6, 8, 4)

II

Plantactinospora

m-DAP

Ara, Xyl

3b

9 (H4), 10 (H4)

II

Polymorphospora

m-DAP

Xyl

2a

9 (H4, 6), 10 (H4, 6)

II

Rugosimonospora

3-OH-DAP

Ara, Gal, Xyl

2c

9 (H8, 6)

II

Salinispora

m-DAP

Ara, Gal, Xyl

3a

9 (H4)

II

Spirilliplanes

m-DAP

Ara, Xyl

2d

10 (H4)

II

Verrucosispora

m-DAP

Man, Xyl

2d

9 (H4)

II

Virgisporangium

m-DAP

Ara, Gal, Man, Rha, Xyl

2d

10 (H4, 6, 8)

II

Data for reference genera were taken from Ørskov (1923), Couch (1950), Kane (1966), Thiemann et al. (1967), Asano and Kawamoto (1986), Yokota et al. (1993), Rheims et al. (1998), Kudo et al. (1999), Tamura et al. (1994, 1997, 2001, 2006), Lee and Hah (2002), Matsumoto et al. (2003), Maldonado et al. (2005), Thawai et al. (2006, 2010), Ara and Kudo (2006, 2007a, b), Ara et al. (2008a, b), Wiese et al. (2008), Monciardini et al. (2009), Qin et al. (2009), Inahashi et al. (2010), Lee and Lee (2011), Xie et al. (2011) and Li et al. (2011). Fatty acid types are classified according to Kroppenstedt (1985) and phospholipid types according to Lechevalier et al. (1981)

m-DAPmeso-diaminopimelic acid, l-Lysl-lysine, Ara arabinose, Gal galactose, Glc glucose, Man mannose, Rha rhamnose, Rib ribose, Xyl xylose

On the basis of morphological, chemotaxonomic data, phylogenetic analysis and the signature nucleotide pattern of the 16S rRNA gene, strain NEAU-J3T is readily distinguishable from known actinomycetes belonging to the family Micromonosporaceae and thus this isolate can be considered as representative of a new genus. Therefore, we propose that strain NEAU-J3T represents a novel species of a new genus within the family Micromonosporaceae, for which the name W. harbinensis gen. nov., sp. nov. is proposed.

Description of Wangella gen. nov.

Wangella (Wang. ell’a. N.L. fem. Dim. n. Wangella, named after Jidong Wang, a Chinese microbiologist).

Aerobic, Gram-positive and non-acid-fast actinomycetes. Cells form extensively branched, non-fragmenting, yellow- to rusty-coloured substrate mycelium on ISP3 agar. Aerial hyphae and sporangia are not present. Spores (1.60 × 0.80 μm) are oval and non-motile with a smooth surface and are borne singly on the substrate mycelium. Cell walls contain meso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars are galactose, ribose and glucose. The acyl type of the cell wall polysaccharides is glycolyl. The predominant menaquinones are MK-9 (H4) and MK-9 (H6). The phospholipids are phosphatidylcholine and phosphatidylethanolamine, corresponding to phospholipid type PIII. The major cellular fatty acids are C16:0, C18:1 ω9c, C18:0, C17:0, C17:1 ω7c, anteiso-C17:0, C16:1 ω7c, and C15:0, corresponding to fatty acid type 1a. The type species of the genus is W. harbinensis.

Description of Wangella harbinensis sp. nov.

Wangella harbinensis (har.bin.en′sis. N.L. masc. adj. harbinensis pertaining to the city in Heilongjiang Province in the north of China, from where the type strain was isolated).

Morphological, chemotaxonomic and general characteristics are as given above for the genus. Grows well on oatmeal-nitrate, tap-water, yeast extract-starch, glucose-yeast extract, ISP2, ISP3, ISP4 and ISP7 agar; moderately well on glucose-asparagine, Bennett’s and HV agar; poorly on sucrose-nitrate and ISP6 agar; no growth is observed on Hickey–Tresner agar and ISP5 medium. No soluble pigments are produced on any of these media. Positive for liquefaction of gelatin and negative for hydrolysis of starch, decomposition of cellulose, reduction of nitrate and production of H2S. d-Glucose, d-galactose, d-mannose, d-maltose, d-sucrose, l-rhamnose, d-sorbitol and d-raffinose are utilized as sole carbon sources but l-arabinose, d-fructose, inositol, lactose, d-mannitol, d-ribose and d-xylose are not. l-Arginine, l-threonine, l-alanine, l-glutamine, l-asparagine, creatine and glycine are used as sole nitrogen sources but l-aspartic acid, l-glutamic acid and l-tyrosine are not. Tolerates up to 3 % (w/v) NaCl and grows at temperatures between 18 and 39 °C, with an optimum temperature of 28 °C. Growth occurs at initial pH values between 4 and 10, the optimum being pH 7.0. The G + C content of the genomic DNA is 62.5 ± 0.25 mol %.

The type strain is NEAU-J3T(=CGMCC 4.7039T = DSM 45747T), isolated from soybean root collected from Harbin, Heilongjiang Province, north China. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain NEAU- J3T is JQ750973.

Acknowledgments

This work was supported in part by grants from NKPBR (No. 2010CB126102) and NNSF (Nos. 30971937 and 30771427) of China, the Special Foundation for Scientific and Technological Innovation Research of Harbin (No. 2011RFXXN038), NSF of Heilongjiang Province (No. C201029).

Supplementary material

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© Springer Science+Business Media Dordrecht 2012