Archives of Microbiology https://doi.org/10.1007/s00203-018-1530-2
ORIGINAL PAPER
Paraburkholderia panacihumi sp. nov., an isolate from ginsengcultivated soil, is antagonistic against root rot fungal pathogen Yue Huo1 · Jong‑Pyo Kang1 · Yeon‑Ju Kim1 · Deok‑Chun Yang1,2 Received: 26 December 2017 / Revised: 27 April 2018 / Accepted: 24 May 2018 © Springer-Verlag GmbH Germany, part of Springer Nature 2018
Abstract The novel species DCY115T was isolated from ginseng-cultivated soil in Gochang province, Republic of Korea. The isolated strain was assigned to the genus Paraburkholderia due to its 16S rRNA gene sequence proximity to Paraburkholderia xenovorans LB400T (98.8%), Paraburkholderia terricola LMG 20594T (98.4%), Paraburkholderia graminis C4D1MT (98.2%), Paraburkholderia rhynchosiae WSM3937T (98.1%), and Paraburkholderia phytofirmans PsJNT (98.1%). Strain DCY115T is gram-negative, facultative aerobic, rod-shaped, non-motile, non-flagellated, and oxidase and catalase positive. The predominant isoprenoid quinone of D CY115T is ubiquinone Q-8. The major cellular fatty acids are C16:0, cyclo-C17:0, cyclo-C19:0 ω8c, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The major polar lipids include diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and an unknown amino lipid (AL1). The genomic DNA G + C content is 61.3 mol%. Phenotypic tests and chemotaxonomic analysis place strain D CY115T in the genus Paraburkholderia. DNA–DNA hybridization values between strain DCY115T and closely related reference strains were lower than 51%. The low DNA relatedness data in combination with phylogenetic and biochemical tests showed that strain DCY115T could not be assigned to any recognized species. Finally, strain DCY115T showed antagonistic activity against Fusarium solani (KACC 44891T) and Cylindrocarpon destructans (KACC 44660T), which are two root rot fungal pathogens of ginseng. In conclusion, the results in this study support strain D CY115T as a novel species within the genus Paraburkholderia for which the name Paraburkholderia panacihumi is proposed. The type strain is DCY115T (= KCTC 52952T = JCM 32099T). Keywords Taxonomy · Paraburkholderia panacihumi · Ginseng soil · Antagonistic activity
Introduction Communicated by Erko Stackebrandt. Yue Huo and Jong-Pyo Kang contributed equally to this work. The NCBI GenBank accession number for the 16S rRNA gene sequence of strain DCY115T is KY694400. DPD (digital protologue database) Taxon Number TA00399. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00203-018-1530-2) contains supplementary material, which is available to authorized users. * Yeon‑Ju Kim
[email protected] * Deok‑Chun Yang
[email protected] Extended author information available on the last page of the article
The genus Burkholderia belongs to the family Burkholderiaceae of the Betaproteobacteria as described by Yabuuchi et al. (1992). However, phylogenetic studies based on 16S rRNA and several housekeeping genes showed that the genus Burkholderia was not monophyletic and that it contains at least two genera: the genus Burkholderia containing clinically important and phytopathogenic members of the genus and a new genus, Paraburkholderia gen. nov., harboring environmental species (Dobritsa and Samadpour 2016). The genus Paraburkholderia was first proposed by Sawana et al. (2014) with Paraburkholderia graminis as the type species. Members of the genus Paraburkholderia are characterized as gram-negative, straight rod-shaped, and with a G + C content in the range 58.9–65.0 mol% (Dobritsa and Samadpour 2016). At the time of writing, the genus Paraburkholderia is comprised of 65 recognized species
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(http://www.bacterio.net/paraburkholderia.html). Members of the genus Paraburkholderia have been isolated from various environments (soil, plants and water), including the recently described Paraburkholderia caffeinilytica (Gao et al. 2016) and Paraburkholderia azotifigens (Choi and Im 2018). Ginseng (Panax Ginseng Meyer) has been used in traditional medicine in East Asia for thousands of years. The main active compounds, derived from glycosylated triterpenes or ginsenosides, possess many pharmacological properties (Choi 2008). Despite its prevalent and widespread usage, ginseng cultivation is hindered by root rot, a major disease caused by the soil-born fungi Cylindrocarpon destructans and Fusarium solani (Rahman and Punja 2005). Accordingly, screening of D CY115T isolated from ginsengcultivated soil was carried out by in vitro studies to confirm antifungal activity against F. solani (KACC 4 4891T) and C. destructans (KACC 44660T). We isolated strain DCY115T and identified it as the type strain of novel species in the genus Paraburkholderia. Accordingly, strain DCY115T is properly identified as a novel Paraburkholderia species, for which the name Paraburkholderia panacihumi sp. nov. is propounded.
Materials and methods Isolation and culture conditions Rhizosphere soil samples were collected from a ginseng field in Gochang county (35°26′89″N 126°42′740″E), Republic of Korea. A 1.0 g soil sample was dissolved in 10 ml sterile saline (0.85% NaCl in distilled water, w/v). The mixture was serially diluted up to a 1 0−5 dilution and 100 µl of each dilution was spread-plated onto tryptic soy agar (TSA, MB cell) media. The plates were incubated at 30 °C for 3 days. Colonies with different morphology, color, and margin were picked and purified by transferring to new TSA plates. Isolated and purified colonies were sent to GenoTech (Daejeon, Republic of Korea) for 16S rRNA sequencing. Among the purified isolates, a novel Paraburkholderia species was identified and named D CY115T, and was characterized using a polyphasic approach. Tryptic soy broth (TSB, MB cell) containing 30% (v/v) glycerol was used as storage medium and the strain was kept at − 80 °C for long-term maintenance. Strain D CY115T was deposited to the Korean Collection for Type Cultures (KCTC 5 2952T) and the Japan Collection of Microorganisms (JCM 32099T). Phylogenetically closely related type strains were purchased for comparative analysis: P. xenovorans (DSM 1 7367T) and P. graminis (DSM 17151T) were obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) collection; P. terricola (LMG 20594T), P. rhynchosiae (LMG 2 7174T), and P. phytofirmans (LMG 2 2487T)
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were obtained from the Belgian Coordinated Collections of Microorganisms (BCCM). Additionally, F. solani (KACC 44891T) and C. destructans (KACC 44660T) were obtained from the Korean Agricultural Culture Collection (KACC) to confirm antifungal activity.
16S rRNA sequencing and phylogenetic construction The 16S rRNA gene was amplified using the 27F/1492R (Lane 1991) and 518F/800R universal bacterial primer sets (Weisburg et al. 1991). Seq-Man software version 4.1 (DNASTAR, Inc.) was utilized to compile and edit the 16S rRNA sequence of strain DCY115T. Alignment gaps were manually trimmed using the BioEdit program (Hall 1999). Multiple alignments were conducted using the CLUSTAL X program (Thompson et al. 1997). Evolutionary distances were calculated according to the Tamura–Nei model (Tamura and Nei 1993). Finally, the phylogenetic tree was constructed using neighbor-joining- (Saitou and Nei 1987), maximum-parsimony- (Fitch 1971), and maximum-likelihood (Felsenstein 1981) methods using the MEGA 6 program package (Tamura et al. 2013). Bootstrap values were determined based on 1000 replications (Felsenstein 1985). The 16S rRNA gene sequence of strain DCY115T was compared with sequences listed in the public database using the EzTaxon-e server [http://eztaxon-e.ezbiocloud.net/; (Kim et al. 2012)].
Phenotypic characteristics analysis Colonies of D CY115T grown on TSA media were monitored after incubation at 30 °C for 3 days. Cell morphology was visually analyzed by transmission electron microscopy (TEM). Cells cultured on TSA media at 30 °C for 24 h were suspended and placed on carbon and formvar-coated nickel grids for 30 s. The grids were coated with a drop of 0.1% (w/v) aqueous uranyl acetate to aid in observation. Colonies were observed under a microscope operated at 80 kV (Carl Zeiss LOE912AB) under standard operating conditions. Gliding motility was tested using a hanging-drop technique (Bernardet et al. 2002). A Gram stain kit (bioMérieux) was used to verify the gram reactivity. Oxidase activity was detected by 1% (w/v) N,N,N,N-tetramethyl-p-phenylenediamine reagent (bioMérieux) as instructed by the manufacturer. Catalase activity was analyzed by the production of O2 bubbles from freshly grown cells mixed with a 3% (v/v) H2O2 solution. The ability of strain D CY115T to grow under anaerobic conditions was determined using the GasPak™ EZ Gas Generating System (Becton Dickinson) after 2 weeks of incubation at 30 °C. Growth was tested using several media, including Reasoner’s 2A (R2A, MB cell) agar, Trypticase soy agar (TSA, MB cell), lysogeny broth agar (LB, MB cell),
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nutrient agar (NA, Difco), potato dextrose agar (PDA, MB cell), and MacConkey agar (Difco) at 30 °C for 1 week. Salinity tolerance was investigated using TSB supplemented with 0–5.0% (w/v) NaCl, at 0.5% intervals, after incubation at 30 °C for 5 days. The temperature values of 4, 10, 15, 20, 25, 30, 35, 37 and 40 °C were tested for 1 week. Growth at pH 4.0–10.0 in pH 0.5-unit intervals was evaluated in TSB broth. The pH level was adjusted using methods previously described by Kang et al. (2015). After 5 days of incubation at 30 °C, measurement of optical density at 600 nm was performed by UV–Vis spectrophotometry (Ultrospec 2100 Pro, Amersham Biosciences). Hydrolysis of the following substrates was analyzed: tyrosine [on R2A agar containing 0.5% (m/v) tyrosine], casein [on R2A agar supplemented with 2 % (m/v) skim milk], Tween 80 [on R2A agar containing 1% (v/v) Tween 80 and 0.02% (m/v) C aCl2], DNA (on DNase agar medium) and gelatin [on TSA agar containing 1.2% (m/v) gelatin]. H 2S production was analyzed using the methods previously described by Levine et al. (1934). Antibiotic susceptibility was tested on Mueller–Hinton (Difco) agar at 30 °C for 48 h using the disk diffusion method (Bauer et al. 1966). The inhibition zones were interpreted according to the manufacturer’s instructions. The antibiotic disks used were as follows: neomycin (30 µg), vancomycin (30 µg), tetracycline (30 µg), lincomycin (15 µg), penicillin G (10 U), erythromycin (15 µg), rifampicin (5 µg), novobiocin (30 µg), ceftazidime (30 µg), oleandomycin (15 µg), and carbenicillin (100 µg). Other phenotypic analyses were performed with the commercial API ZYM, API 20NE, and API ID 32GN kits (bioMérieux) as instructed by the manufacturer.
Determination of DNA G + C content The genomic DNA G + C mol% content was analyzed with modification as described by Mesbah et al. (2011). The nucleosides were detected by HPLC (NS-4000, Futecs co. Ltd.), and separated using a reverse-phase YMC-Triart C18 column (250 × 4.6 mm, 5 µm). The elution solution was a mixture of 25 mM ammonium dihydrogen phosphate (NH4)H2PO4/acetonitrile (20/1, v/v). The flow rate was 0.6 ml min−1. Detection was performed by UV absorption at a wavelength of 270 nm. The genomic DNA of Escherichia coli D4889 (Sigma-Aldrich) was used as a calibration standard. Experiments were carried out in triplicate.
DNA–DNA relatedness DNA hybridization was performed fluorometrically using photobiotin-labeled DNA probes and microdilution wells as reported previously by Ezaki et al. (1989) with slight modifications. The optimal hybridization temperature was estimated to be 49.4 °C. The experiment was performed five times for each sample to ensure accuracy of the results.
Chemotaxonomic analysis Polar lipids of strain DCY115T and P. xenovorans LB400T were extracted and subsequently analyzed by two-dimensional thin layer chromatography (TLC) on 10 × 10 cm silica gel plates (Merck No. 5553) as described by Minnikin et al. (1984). Ubiquinones were extracted from dry cells with chloroform: methanol (2/1, v/v), concentrated at 50 °C in a vacuum rotary evaporator, and the residues were extracted with hexane. After separation of the ubiquinones using a Sep-Pak Vac 6 ml cartridge, hexane/diethyl ether (90/10, v/v) was used for elution. Following evaporation, ubiquinones were dissolved in acetone and concentrated using N 2 gas. Finally, ubiquinones were analyzed by HPLC [model, NS-6000A, Futecs; reversed-phase column YMC-Triart C18 (250 × 4.6 mm × 0.5 µm), a flow-rate of 1.0 ml/min and wavelength 270 nm; solvent methanol/2propanol (65/35, v/v)] according to the method of Hiraishi et al. (1996). Fatty acids were saponified, methylated, and extracted from cells cultured on TSA at 30 °C for 48 h according to the MIDI standard protocol (Sherlock Microbial Identification System) as previously reported by Sasser (1990) and then analyzed by gas chromatography (Agilent GC 6890). Identification of methyl esters was analyzed using the TSBA library (version 6.1).
In vitro assessment of plant growth promotion and/ or antagonism Siderophore production was detected using Pseudomonas agar F medium with chrome azurol S (CAS) complex, as described by Schwyn and Neilands (1987). Siderophore production is confirmed by the change in color of a yellowish to reddish halo surrounding the colonies in the blue-green-colored medium. Phosphate solubilizing ability was verified by a plate screening method developed by Pikovskaya (1948). A clear halo region found around the colonies in the opaque medium indicated a positive phosphate solubilization result. The antagonistic ability of D CY115 T against the ginseng root rot fungal pathogens F. solani (KACC 4 4891T) and C. destructans (KACC 44660T) was investigated with the methods described by Farh et al. (2015). All pathogenic fungi were activated on PDA medium and incubated at 25 °C for 5 days. Strain DCY115T and P. xenovorans LB400T were activated on TSA and incubated for 2 days at the optimum temperature of 30 °C. All bacteria and fungi were tested for antagonistic ability on Waksman agar. Pathogen growth was verified at different time points (after 10 days for F. solani; after 20 days for C. destructans) and compared with the control.
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Results and discussion
Physiological and biochemical characteristics
Phylogenetic analysis
Cells are rod shaped (1.8–2.0 µm in length × 0.7–0.8 µm in width) and non-flagellated (Supplementary Fig. S2). Physiological and biochemical characteristics of strain DCY115T are generalized in the species description and the comparison of selective characteristics with related type strains is presented in Table 1. Strain DCY115T can be differentiated from strain P. xenovorans LB400T by the following characteristics. The API ZYM strip test indicated that D CY115T is esterase (C4) and cystine arylamidase positive, the API 20 NE strip indicated capric acid, d-glucose, l-arabinose, potassium gluconate, trisodium citrate, phenylacetic acid-positive activity, and the API ID 32 GN strip indicated d-ribose, 3-hydroxybenzoic acid, and l-fucose-positive results.
Strain D CY115 T showed the highest 16S rRNA gene sequence similarity with P. xenovorans LB400T (98.8%), P. terricola LMG 2 0594T (98.4%), P. graminis C4D1MT (98.2%), P. rhynchosiae WSM3937 T (98.1%), and P. phytofirmans PsJNT (98.1%). The phylogenetic tree constructed using the maximum likelihood (Fig. 1), neighbour joining (Supplementary Fig. S1), and maximum parsimony algorithms indicated that strain DCY115T belongs to the genus Paraburkholderia and forms a monophyletic cluster with P. xenovorans LB400T.
Fig. 1 16S rRNA gene sequence maximum-likelihood phylogenetic tree showing the taxonomic position of strain D CY115T in the genus Paraburkholderia. Filled circles indicate that the corresponding nodes were also recovered in trees generated with maximum-
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parsimony algorithms. Bootstrap values > 50% based on 1000 replications are shown at branching points. Pelistega europaea LMG 10982T (Y11890) was used as the outgroup. Bar, 0.01 substitutions per nucleotide position
Archives of Microbiology Table 1 Different characteristics of strain DCY115T and related Paraburkholderia-type strains
Characteristics
1
Hydrolysis of Esculin − DNA − Enzyme activity (API ZYM) Esterase − Lipase + Cystine arylamidase − β-galactosidase − β-glucosidase − Reduction of nitrates − Assimilation of (API 20NE, ID 32 GN) + l-arabinose Potassium gluconate − Capric acid − Adipic acid − Trisodium citrate − Phenylacetic acid − − l-rhamnose − d-ribose − d-saccharose Sodium malonate – Sodium acetate – 3-hydroxybenzoic acid + − l-serine – l-fucose Propionic acid − 4-hydroxybenzoic acid +
2
3
4
5
6
− −
− −
− +
+ −
− −
+ + + − − −
− − − + − +
+ − − − − +
+ − − − + −
+ + − − − −
− + + − + + − + − – – – − + − +
+ + + − + − − + + + – – + + +
+ + + + + + + − + + + – + + – –
+ + − − + + + − + + + + + + + +
+ + + + − + + + – – + + + + + +
Strains: 1, strain D CY115T; 2, Paraburkholderia xenovorans DSM 17367T; 3, Paraburkholderia terricola LMG 2 0594T; 4, Paraburkholderia graminis DSM 17151T; 5, Paraburkholderia rhynchosiae LMG 27174T; 6, Paraburkholderia phytofirmans LMG 2 2487T. All data were from this study, +, positive; −, negative
DNA G + C content and DNA–DNA relatedness The genomic DNA G + C content is estimated to be 61.3 mol%. The DNA–DNA hybridization value between strain DCY115T and P. xenovorans LB400T was estimated to be 51.0%, and DNA–DNA hybridization values between strain DCY115T and other strains were well below 31.1%. The low hybridization values suggest that DCY115T represents a distinct genomic species.
Chemotaxonomic characteristics T
The polar lipid profiles of strain DCY115 and P. xenovorans LB400T are listed in Supplementary Fig. S3. The predominant isoprenoid quinone of DCY115T is ubiquinone Q-8, which is in accordance with the description of the genus Paraburkholderia. The fatty acids profiles of strain DCY115T and the related type strains are listed in Table 2. The major cellular fatty acids are C16:0, cyclo-C17:0,
16:1 cyclo-C19:0 ω8c, summed feature 3 ( C16:1 ω7c and/or C ω6c), and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). Overall, the chemotaxonomic results show that strain D CY115 T has the typical characteristics of the genus Paraburkholderia. However, the different proportion of fatty acids revealed that DCY115T was distinguishable from related-type strains as compared in this study.
Plant growth promoting characteristics and antagonistic activity Regarding the plant growth promoting characteristics, strain DCY115T and P. xenovorans LB400T showed the ability to produce siderophores as indicated by the change in color of a yellow halo zone surrounding colonies in the blue-greencolored medium (Fig. 2a). Moreover, strain D CY115T and P. T xenovorans LB400 produced a clear halo region around the colonies in Pikovskaya medium, indicating a positive result for phosphate solubilization (Fig. 2b). DCY115T and P.
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Table 2 Fatty acid profile of strain DCY115T and relatedtype strains
Archives of Microbiology Fatty acids
1
2
3
4
5
6
C14:0 C16:0 Cyclo-C17:0 C16:1 2-OH C16:0 2-OH C16:0 3-OH C18:1 2-OH Cyclo-C19:0 ω8c Summed feature 2* Summed feature 3** Summed feature 8***
3.4 20.67 10.32 1.86 1.81 4.6 – 12.22 6.88 12.33 24.82
4.7 18.2 5.1 2.2 2.2 7.1 tr 3.6 8.5 19.1 27.3
4.8 14.3 14 3.1 3.5 6.6 1.5 7.1 8.2 9.1 27.8
4.7 14.5 6.9 5.3 3.7 7 1.8 3.3 9.8 15.3 26.5
3.7 14.4 3.5 2.7 1.5 5.4 – 1.7 6.4 20.9 38.2
3.6 13.8 2.1 2.3 2.1 4.1 1.4 1.8 5.6 17.6 44.3
Strains: 1, strain D CY115T; 2, Paraburkholderia xenovorans DSM 17367T; 3, Paraburkholderia terricola LMG 2 0594T; 4, Paraburkholderia graminis DSM 17151T; 5, Paraburkholderia rhynchosiae LMG 27174T; 6, Paraburkholderia phytofirmans LMG 22487T. All data were obtained in this study − not detected, tr trace amount (< 1.0%) *Summed feature 2 comprising C12:0 aldehyde, unknown fatty acid of equivalent chain length 10.928 and/ or C14:0 3-OH and/or C 16:1 iso I 16:1 ω6c **Summed feature 3 comprising C16:1 ω7c and/or C
18:1 ω6c ***Summed feature 8 comprising C18:1 ω7c and/or C
xenovorans LB400T showed an antagonistic activity against F. solani (KACC 44891T). However, DCY115T showed an antagonistic activity against C. destructans (KACC 4 4660T) (Fig. 2c–h). Overall, the results collected during this study indicate that strain DCY115T belongs to a new species within the genus Paraburkholderia. We suggest the name Paraburkholderia panacihumi sp. nov. for this isolate from ginsengcultivated soil.
Description of Paraburkholderia panacihumi sp. nov Paraburkholderia panacihumi (pa.na.ci.hu’mi. N.L. n. Panaxacis scientific name of ginseng; L. n. humus soil; N.L. gen. n. panacihumi of soil of a ginseng field). Cells are gram-negative, catalase and oxidase positive, facultative anaerobic, rod shaped approximately 0.7–0.8 µm in width and 1.8–2.0 µm in length. Colonies on TSA medium are circular with entire margins, slightly yellow pigmentation and 0.4–1 mm in diameter after 3 days of incubation at 30 °C. Growth occurs at 10–37 °C (optimum 30 °C), at pH 4.5–7.5 (optimum pH 6.5) and in the presence of 0–1.5% (w/v) NaCl (optimum 0.5%). Growth occurs on NA, R2A, TSA, PDA, and LB agar medium. Hydrolysis of Tween 80 is positive. Casein, DNA, tyrosine, H2S production, gelatin, and urease are negative. In the API ZYM strip, alkaline phosphatase, esterase lipase (C8), lipase (C14), leucine arylamidase, valine arylamidase, acid phosphatase and
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naphthol-AS-BI-phosphohydrolase activity is positive. In the API 20 NE strips, the assimilation of d-glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine and malic acid are positive In the API ID 32 GN strips, the assimilation of myo-inositol, lactic acid, l-alanine, 3-hydroxybenzoic acid, d-sorbitol, l-histidine, potassium 2-ketogluconate, 3-hydroxybutyric acid, 4-hydroxybenzoic acid, and l-proline activities are positive. Negative characteristics from commercial test systems are listed in Supplementary Table S1. Strain DCY115T was susceptible to neomycin (30 µg), tetracycline (30 µg), penicillin G (10 U), erythromycin (15 µg), novobiocin (30 µg), ceftazidime (30 µg), and carbenicillin (100 µg) but resistant to vancomycin (30 µg), lincomycin (15 µg), rifampicin (5 µg), and oleandomycin (15 µg). The genomic DNA G + C content is 61.3 mol%. The major polar lipids are diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and an unknown amino lipid (AL1). The predominant isoprenoid quinone of DCY115T is ubiquinone Q-8. The major cellular fatty acids consist of C16:0, cyclo-C17:0, cyclo-C19:0 ω8c, summed feature 3 (C16:1 ω7c and/or C 16:1 ω6c) and summed feature 8 (C18:1 ω7c and/or C 18:1 ω6c). Siderophores are produced and phosphate is solubilized in this strain. D CY115T shows an antagonistic effect against the ginseng root rot pathogens F. solani (KACC 44891T) and C. destructans (KACC 44660T). The type strain, D CY115T (= KCTC 52952T = JCM 32099T) was isolated from ginseng-cultivated soil in Gochang province, Republic of Korea.
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Fig. 2 In vitro assessment of plant growth in the presence of strain DCY115T and Paraburkholderia xenovorans DSM17367T. Siderophore production ability (a); phosphate-solubilizing ability (b); antagonistic ability (c–h); F. solani KACC 4 4891T (c–e); control (c);
P. xenovorans D SM17367T (d); DCY115T (e); C. destructans KACC 44660T (f–h); control (f); P. xenovorans DSM17367T (g); DCY115T (h)
Acknowledgements This study was supported by a Grant from the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (KIPET No. 317007-3), Republic of Korea.
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Archives of Microbiology
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Affiliations Yue Huo1 · Jong‑Pyo Kang1 · Yeon‑Ju Kim1 · Deok‑Chun Yang1,2 1
Department of Oriental Medicinal Biotechnology, College of Life Science, Kyung Hee University, Seocheon‑dong, Giheung‑gu, Yongin, Gyeonggi 446‑701, Republic of Korea
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2
Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University, Seocheon‑dong, Giheung‑gu, Yongin, Gyeonggi 446‑701, Republic of Korea
