• Korean Journal of Microbiology
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• v.54 no.4
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• pp.471-473
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• 2018

Variovorax sp. PMC12 is a rhizobacterium isolated from tomato rhizosphere and enhanced the plant resistance to abiotic and biotic stresses. Here we present the complete genome sequence of strain PMC12. The genome is comprised of two circular chromosomes harboring 5,873,297 bp and 1,141,940 bp, respectively. A total of 6,436 protein-coding genes, 9 rRNAs, 64 tRNAs, 3 ncRNAs, and 80 pseudogenes were identified. We found genes involved in 1-aminocyclopropane-1-carboxylate (ACC) deaminase, antioxidant activity, phosphate solubilization, and biosynthesis of proline and siderophore. Those genes may be related to capability of improving plant resistance to various stresses including salinity, cold temperature, and phytopathogen.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.90.1-90
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• 2003

Colonization of Pseudomonas chlororaphis O6, a nonpathogenic rhizobacterium, on the roots induced systemic resistance in cucumber plants against tai-get leaf spot, a foliar disease caused by Corynespora cassiicola. A cDNA library was constructed using mRNA extracted from the cucumber leaves 12 h after inoculation with C. cassiicola, which roots had been previously treated with O6. To identify the genes involved in the O6-mediated induced systemic resistance (ISR), we employed a subtractive hybridization method using mRNAs extracted from C cassiicola-inoculated cucumber leaves with and without previous O6 treatment on the plant roots. Differential screening of the cDNA library led to the isolation of 5 distinct genesencoding a GTP-binding protein, a putative senescence-associated protein, a galactinol synthase, a hypersensitive-induced reaction protein, and a putative aquaporin. Expressions of these genes are not induced by O6 colonization alone. Before challenge inoculation, no increase in the gene transcriptions could be detected in previously O6-treated and untreated plants but, upon subsequent inoculation with the pathogenic fungus, transcription levels in O6-treated plants rose significantly faster and stronger than in untreated plants. Therefore, the O6-mediated ISR may be associated with an enhanced capacity for the rapid and effective activation of cellular defense responses which becomes apparent only after challenge inoculation on the distal, untreated plant parts, as suggested by Conrath et al. (2002). This work was supported by a grant R11-2001-092-02006-0 from the Korea Science and Engineering Foundation through the Agricultural Plant Stress Research Center at Chonnam National University.

• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.607-620
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• 2023

The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.32-32
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• 2023

Cadmium and salt exposure to crops is considered vulnerable for production as well as consumption. To address these challenges, the current study aimed to mitigate the toxicity induced by salt and cadmium in soybean plants through the application of bacterial strain Bacillus safensis KJW143 isolated from the rhizosphere of oriental melon..The bioassay analysis revealed that KJW143 is a highly salt-tolerant and cadmium-resistant (Cd) strain with an innate ability to produce melatonin, gibberellin (GA3), Indole-3-Acetic Acid (IAA), and organic acids (i.e., acetic, succinic, lactic, and propionic acids). Soybean plants at 20 days old were treated with KJW143 in a different form (pellet, broth, and together) and their effect on plant performance was investigated. Inoculation with KJW143enhanced plant biomass and growth attributes in soybean plants compared to the control (non-treated). In particular, we observed that only pellet-treated showed 65%, 27.5%, and 28.7% increase in growth (shoot fresh weight) compared to broth, broth with pellet, and control. In addition, bacterial strain KJW143 treatment (only pellet) modulated the physiochemical apparatus of soybean plants by increasing glucose (390%), arabinose (166%), citric acid (22.98%) and reducing hydrogen peroxide (29.7%), catalase (32.1%), salicylic acid (25.6%) compared to plants with combined stressed plants (cd and salinity). These findings suggest that bacterial strain KJW143 could be usedas a biofertilizer to minimize the probable risk of heavy metal and salinity stress on crops.

• Research in Plant Disease
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• v.30 no.2
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• pp.114-123
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• 2024

In July 2022, stem rot symptom was found in a peony plant grown in a pot under a greenhouse at Jinju, Gyeongnam Province, South Korea. Two fungal species were isolated from the infected peony stems and cultured on 1/2-strength potato dextrose agar for identification. The morphological characteristics of the fungal isolates were examined, and nucleotide sequences of the internal transcribed spacer region, β-tubulin and translation elongation factor 1-α were analysed. The pathogenicity of the two isolates was confirmed in detached peony leaves, according to Koch's postulates. To our knowledge, this is the report of Neopestalotiopsis clavispora and Sclerotinia sclerotiorum as the causal agents of peony stem rots. Antifungal activity of chemical fungicide propineb and rhizobacterium Bacillus siamensis H30-3 was shown against the two plant pathogenic fungi N. clavispora and S. sclerotiorum.Unidentified diffusible and volatile compounds from B. siamensis H30-3 could suppress in vitro mycelial growths of N. clavispora JJ 8-2-1 and S. sclerotiorum JJ 8-2-2.

• Journal of Life Science
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• v.13 no.1
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• pp.90-98
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• 2003

An allium rhizobacterium Serratia plymuthica AL-1 was previously selected as a biocontrol agent of allium white rot. The chitinase from S. plymuthica AL-1 produced in medium containing colloidal chitin was purified by ammonium sulfate precipitation (40~70%), affinity adsorption, column chromatography on DEAE-sephadex A-50 and sephadex C-200 gel filtration. The enzyme was purified 10.8-fold with a yield of 7.3% from the starting culture broth. The purified chtinase gave a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis, it's molecular weight was estimated to be 55 kDa. The optimum pH and temperature of the purified enzyme were pH 5.5 and $55^{\circ}C$, respectively and it is stable up to $50^{\circ}C$ and maintains around 90% of its activity for 60min. The enzyme were activated by $Ca^{2+}$, $Mn^{2+}$ and $Mg^{2+}$ and inhibited by $Cu^{2+}$, SDS, $\rho$-CMB, MIA, respectively. The purified chitinase showed broad spectrum of antifungal activities against plant pathogenic fungi Sclerotium cepivoruin, Alternana alternnta, Colletotrichum glceosporioidrs, Phoma sp., Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium f. sp. niveum but rarely inhibited Phytophthora capsici and Pythium ultimum.. The purified chitinase from S. plymuthica AL-1 caused swelling, lysis, deceleration and degradation of the hyphal tips of S. sczerotiorum causing allium white rot. It suggest that S. prymuthica AL-1 chitinase play an important part in the bifunctional chitinase / lysozyme activity.

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.1-9
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• 2009

The genetic monitoring method was developed for the rapid detection of the PGPR and biocontrol agent, B. subtilis AH18 in red-pepper field soil by multiplex PCR using sid, aec and cel gene primers. The monitoring of B. subtilis AH18 in the soil was carried by amplified a 2,3-dihydro-2,3-dihydroxy benzoate dehydrogenase [EC: 1. 3. 1. 28]gene (sid - 794 bp : EF408238) which is a key enzyme of siderophore synthesis, an auxin efflux carrier gene (aec - 1,052 bp : EF408239) and a cellulase gene (cel - 1,582 bp : EF070194). The natural un sterilized soil was inoculated with B. subtilis AH18 to determine the sensitivity ($1.8\times10^5$ cfu/g) of multiplex PCR for the rapid dectection and then the strain was monitored successfully in rhizosphere or non-rhizosphere soil of red-pepper cultural soil. At 3 weeks after the treatment, density of the strain was monitored more abundantly in rhizosphere soil.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.261-271
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• 2007

Phytoremediation is an economic and environmentally friendly technique to remediate contaminated-soil. In this study, the effects of plants, rhizobacteria and physicochemical factors on phytoremediation have been reviewed. For successful phytoremediation, the selection of plants is primarily important. To remediate soil contaminated with petroleum hydrocarbon, raygrass (Lolium multiflorum lam), white mustard, vetch (Vicia villosa), tall fescue (Festuca arundinacea), legumes, poplar, and Pine (Pinus densiflora) were mainly applied, and the removal efficiency of petroleum hydrocarbon were ranged 68 to 99%. Corn (Zea mays), raygrass (Lolium multiflorum lam), vetch (Vicia villosa), mustard, clover (Trifolium repens), and tall fescue (Festuca arundinacea) were used for the removal of polycyclic aromatic hydrocarbon, and their removal efficiencies were 50-98%. Rhizobacteria play significant roles for phytoremediation because they can directly participate in the degradation of contaminant as well as promoting plants growth. The following rhizobacteria were preferred for phytoremediation: Azospirillum lipoferum, Enterobactor cloacae, Azospirillum brasilense, Pseudomonas putida, Burkholderia xenovorans, Comamonas testosterone, Pseudomonas gladioli, Azotobacter chroococcum, Bacillus megaterium, and Bacillus subtilis. Pysicochemical factors such as pH, temperature, nutrient, electron acceptor, water content, organic content, type of contaminants are consequential limiting factors for phytoremediation.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.17-17
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• 2010

Ethylene, known as a stress hormone regulate wide developmental processes including germination, root hair initiation, root and shoot primordial formation and elongation, leaf and flower senescence and abscission, fruit ripening. The acceleration of ethylene biosynthesis in plant associated with environmental and biological stresses. 1-Aminocycloprophane-1-carboxlyate deaminase(ACCD) is an enzyme that cleaves ACC into and ammonia, a precursor of the plant hormone ethylene. Plant growth-promoting rhizobacteria (PGPR) having ACCD can decrease endogenous ACC level of tissue, resulting in reduced production of ethylene in plants. ACC deaminse was a key enzyme for protect stressed plants from injurious effects of ethylene. ACCD gene was encoded from Pseudomonas flourescens, PGPR and was cloned in Escherichia coli. We expressed the recombinant ACCD(rACCD) containing 357 amino acids with molecular weight 39 kDa that revealed by SDS-PAGE and western blot. The rACCD was purified by Ni-NTA purification system. The active form of rACCD having enzyme activity converted ACC to a-ketobutyrate. The optimal pH for ACC deaminase activity was pH 8.5, but no activity below pH 7.0 and a less severe tapering activity at base condition resulting in loss of activity at over pH 11. The optimal temperature of the enzyme was $30^{\circ}$ and a slightly less severe tapering activity at 15 - 30$^{\circ}$, but no activity over $35^{\circ}$. P. flourescens ACC deaminase has a highly conserved residue that plays in allowing substrate accessibility to the active sites. The enzymatic properties of this rACCD will provide an important reference for analysis of newly isolated ACCD and identification of newly isolated PGPR containing ACCD.