• Applied Biological Chemistry
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• v.50 no.1
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• pp.23-28
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• 2007

Auxin-producing antagonistic bacterium K11, which can inhibit Phytophtora capsici, was isolated from a local red-pepper field soil in Gyeong-buk. In order to check for additional PGPR(plant growth promoting rhizobacterium) functions of the strain K11, we confirmed siderophore and cellulase productions by CAS (chrome azurol S) blue agar and CMC plate with congo red, respectively. The strain K11 was identified as Bacillus licheniformis with 98% similarity on 16s rDNA comparison and Biolog analyses. B. licheniformis K11 promoted mung bean adventitious root induction and enhanced root growth of mung bean (160%), pea (150%), and Chinese cabbage (130%), Also, B. licheniformis K11 was able to effectively suppress (63%) P. capsici causing red-pepper blight in the pot in vivo test. Therefore, we could select a triple-functional PGPR which has auxin, siderophore, and cellulase producing ability for effective crops production in organic farming.

• Journal of Life Science
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• v.17 no.12
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• pp.1682-1688
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• 2007

To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% $Ca_3(PO_4){_2}$ as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from $3{\sim}639{\mu}g/ml$ depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

• Food Science of Animal Resources
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• v.30 no.2
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• pp.328-335
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• 2010

The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• Journal of Life Science
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• v.12 no.3
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• pp.242-249
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• 2002

The optimal culture conditions in 500$m\ell$ flask suture, 5$\ell$ jar fermenter and 2,000 $\ell$ large stale culture were investigated to maximize the production of antibiotic on Rhizoctonia solani AC4, the causal agent of vegetables damping-off, by the strain Bacillus ehimensis YJ-37. Starch (1.5%) as a carbon source, peptone (0.6%) as a nitrogen source and MgC1$_2$(0.15%) as a metal ion in the medium containing N $a_2$HP $O_4$(0.3%) showed the highest production of the antibiotic(s) in a rotary shake (200 rpm). Optimal initial pH of the culture medium, culture temperature and culture time for the antibiotic(s) production were pH 8.0, 32$^{\circ}C$ and 54hrs, respertively. Under the optimal renditions in flask culture, cell growth and antifungal activity (clear zone size) were 1.42 $\times$ 10$^{8}$ cfu/$m\ell$ (21g-DCW/ $\ell$) and 13.9 mm, respectively. In 5 $\ell$ jar fermenter (medium 3 $\ell$), optimal air flow, agitation speed and culture time for the antibiotic(s) production were 2 vvm, 200 rpm and 48hrs, respectively. Under the optimal conditions in 5 $\ell$ jar fermenter, tell growth and antifungal activity were 2.06 $\times$ 10$^{8}$ cfu/$m\ell$ (30g-DCW/ $\ell$) and 13.4 mm, respectively. Under the culture conditions of air flow (30 vvm) and agitation speed (200 rpm) at 32$^{\circ}C$ for 10 days in 2,000 $\ell$ large scale culture (medium 1,800 $\ell$, pH 8.0), cell growth and antifungal activity were 0.81$\times$10$^{8}$ cfu/$m\ell$ (12g-DCW/ $\ell$) and 8.6 mm, respectively.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.447-453
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• 2007

With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

• The Korean Journal of Mycology
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• v.32 no.1
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• pp.31-38
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• 2004

An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

• Journal of Life Science
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• v.18 no.2
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• pp.255-263
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• 2008

Bacteria that antagonize plant pathogenic fungi were isolated from the sediment soil at the Ansan industrial estate. One isolate of them showed growth inhibition of Rhizoctonia solani, Botrytis cenerea, and Fusarium oxysporum. This strain was identified as Pandoraea sp. based on phenotypic and phylogenetic characteristics and termed Pandoraea sp. BCNU 315. Tryptone as nitrogen source and sucrose as carbon source were found to be most effective for the microbial growth. In addition, the optimum temperature and pH for microbial growth were $30^{\circ}C$ and pH 7.0, respectively. The substances generated from Pandoraea sp. BCNU 315 were purified and analyzed by column chromatography, HPLC, GC-MS and NMR. As a result, one compound was determined to be indole, another compound was predicted as cyclopentadecaheptene. Detailed structural clarification of the all of the rest six compounds from Pandoraea sp. BCNU 315 has to be accompanied in the further studies.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.74-80
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• 2011

Bacteriocin-producing lactic acid bacterium having antagonistic activity against Bacillus cereus, was isolated from Kimchi. The selected strain was identified as Lactococcus lactis by the Bergey's manual and 16S rDNA analysis, and named as L. lactis ET45. The bacteriocin was stable in the pH range 3.0-11.0. The bacteriocin was active over a wide temperature range from $40^{\circ}C$ to $121^{\circ}C$. Optimal culture condition for producing bacteriocin was obtained by growing the cells on MRS medium at pH 7.5 and $30^{\circ}C$ for 18 h. Antibacterial activity of the bacteriocin was completely disappeared by proteinase K, and this means that bacteriocin is a proteinous substance. The molecular weight of bacteriocin was estimated to be about 4.5 kDa by tricine sodium dodecyl sulfate polyacryamide gel electrophoresis (TSDS-PAGE).

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1286-1298
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• 2005

The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.