• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.268-275
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• 2008

In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.1-10
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• 2007

The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.36-47
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• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

• Journal of Microbiology and Biotechnology
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• v.30 no.5
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• pp.689-699
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• 2020

Brevibacillus brevis GZDF3 is a gram-positive, plant growth-promoting rhizosphere bacterium (PGPR) isolated from the rhizosphere soil of Pinellia ternata (an important herb in traditional Chinese medicine). The GZDF3 strain produces certain active compounds, such as siderophores, which are the final metabolite products of non-ribosomal peptide synthetase (NRPS) and independent non-ribosomal peptide synthetase (NIS) activity. With the present study, we attempted to investigate the siderophore production characteristics and conditions of Bacillus sp. GZDF3. The antibacterial activity of the siderophores on pathogenic fungi was also investigated. Optimal conditions for the synthesis of siderophores were determined by single factor method, using sucrose 15 g/l, asparagine 2 g/l, 32℃, and 48 h. The optimized sucrose asparagine medium significantly increased the production of siderophores, from 27.09% to 54.99%. Moreover, the effects of different kinds of metal ions on siderophore production were explored here. We found that Fe³⁺ and Cu²⁺ significantly inhibited the synthesis of siderophores. The preliminary separation and purification of siderophores by immobilized-metal affinity chromatography (IMAC) provides strong antibacterial activity against Candida albicans. The synergistic effect of siderophores and amphotericin B was also demonstrated. Our results have shown that the GZDF3 strain could produce a large amount of siderophores with strong antagonistic activity, which is helpful in the development of new biological control agents.

• KSBB Journal
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• v.24 no.4
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• pp.334-340
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• 2009

For the isolation of a plant growth-promoting rhizobacteria, strain YK-5 was selected from approximately 400 thermostable strains isolated from special soil samples. Strain YK-5 produced an antifungal compound, and optimum carbon and nitrogen sources for the production of the antifungal compound were investigated against Aspergillus flavus as a test strain. Modified LB medium containing 1% peptone, 1% yeast extract and 5% black sugar was determined to be the optimal medium for growth and antifungal compound production. Culture broth of strain YK-5 potently inhibited growth of the phytopathogenic fungus Fusarium oxysporum KACC 40052 for 7 days. The plant growth-promotion function of strain YK-5 was tested against radish and rice in pot trials. Leaf number, plant height and root length in YK-5-treated radish markedly exceeded (> 60%) those of untreated radish. Leaf length and white rootlet development were markedly more prominent than in commercially-treated rice plants. Strain YK-5 was determined to be Bacillus subtilis YK-5 by physiological, chemotaxonomical, and phylogenetical analyses.

• Microbiology and Biotechnology Letters
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• v.40 no.3
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• pp.190-196
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• 2012

In this study, a total of more than 1,000 bacteria, including 739 species of aerobic bacteria, 80 species of urease producing bacteria and 303 species of photosynthetic bacteria, were isolated from red-pepper field soils located in the Gyeongsan Province of the Republic of Korea. Amongst these, 158 species of aerobic bacteria, 70 species of urease producing bacteria and 228 species of photosynthetic bacteria were found to be auxin producing soil bacteria through quantification analysis with the Salkowski test. The latter groupings were then tested for antifungal activities to ${\beta}$-Glucanase and siderophore using CMC congo red agar and CAS blue agar media. In addition, the selected strains were examined for antifungal activity against various phytopathogenic fungi on PDN agar media. Six strains; BCB14, BCB17, C10, HA46, HA143, and HJ5, were noted for their ability to both produce auxin and act as antifungal substances. 16S rDNA sequence comparison analyses of these six strains identified them as Bacillus subtilis BCB14, B. methylotrophicus BCB17, B. methylotrophicus C10, B. sonorensis HA46, B. subtilis HA143, and B. safensis HJ5.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.1
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• pp.29-36
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• 2009

Fourteen bacterial strains were isolated from crop rhizosphere and identified as phosphate solubilizing bacteria (PSB) by 16S rRNA analysis. Only 3 strains exhibited a strong ability to solubilize insoluble phosphate in agar medium containing a hydroxyapatite. The rates of P solubilization by isolates were ranged from 200 and $2300\;mg\;L^{-1}$, which are inversely correlated with pH in culture medium. Furthermore, HPLC analyses reveal the production of organic acid from the culture filtrates of PSB. Among these, strain Acinetobacter sp. released only gluconic acid, Pseudomonas orientalis produced gluconic acid which was subsequently converted into 2-ketogluconic acid, and Enterobacter asburiae released acetic acid and succinic acid. On the other hand, P. orientalis and E. asburiae released $372\;mg\;L^{-1}$ and $191\;mg\;L^{-1}$ of IAA into broth culture, respectively, while Acinetobacter sp. did not produce IAA. Furthermore, in vivo study showed that plant growth promoting effect by bacteria generally seemed to be increased IAA production and phosphate solubilization.

• Journal of Life Science
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• v.31 no.7
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• pp.641-653
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• 2021

The purpose of this study was to confirm the antifungal activity, plant growth-promoting activity, and mineral solubilizing activity of 18 types of bacteria isolated purely from rhizosphere soil. The potential of isolates of the genus Bacillus and Pseudomonas as biocontrol agents was confirmed through the antifungal activity of these isolates. This activity has been determined to be due to various hydrolytic enzymes on the cell wall of plant pathogenic fungi and the production of siderophores in isolates. In addition, most of the isolates have been found to have aminocyclopropane-1-carboxylate deaminase production activity, indole-3-acetic acid production activity, and nitrogen fixation activity. These characteristics are believed to have a positive effect on root development, growth, and the productivity of crops via a reduction in the concentration of ethylene under conditions of environmental stress, to which plants are commonly exposed. In addition, on testing for the solubilizing activity of the isolates for phosphoric acid, silicon, calcium carbonate, and zinc, some isolates were found to have mineral solubilizing activities. Inoculation of these isolates during plant growth is expected to assist plant growth by converting nutrients necessary for growth into usable forms that can be absorbed by plants. The 18 isolated strains can be used as biocontrol agents due to their antifungal activity, plant growthpromoting activity, and mineral solubilizing activity.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.391-402
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• 2021

Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

• Research in Plant Disease
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• v.28 no.1
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• pp.1-18
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• 2022

Volatiles exist ubiquitously in nature. Volatile compounds produced by plants and microorganisms confer inter-kingdom and intra-kingdom communications. Autoinducer signaling molecules from contact-based chemical communication, such as bacterial quorum sensing, are relayed through short distances. By contrast, biogenic volatiles derived from plant-microbe interactions generate long-distance (>20 cm) alarm signals for sensing harmful microorganisms. In this review, we discuss prior work on volatile compound-mediated diagnosis of plant diseases, and the use of volatile packaging and dispensing approaches for the biological control of fungi, bacteria, and viruses. In this regard, recent developments on technologies to analyze and detect microbial volatile compounds are introduced. Furthermore, we survey the chemical encapsulation, slow-release, and bio-nano techniques for volatile formulation and delivery that are expected to overcome limitations in the application of biogenic volatiles to modern agriculture. Collectively, technological advances in volatile compound detection, packaging, and delivery provide great potential for the implementation of ecologically-sound plant disease management strategies. We hope that this review will help farmers and young scientists understand the nature of microbial volatile compounds, and shift paradigms on disease diagnosis and management to aromatic (volatile-based) agriculture.