• The Plant Pathology Journal
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• v.23 no.3
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• pp.161-165
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• 2007

Cultivation of a biocontrol agent on a certain medium often results in reduced biocontrol efficacy and alters physiological state. In our previous study, Burkholderia gladioli strain B543 with long-term subculture on tryptic soy agar resulted in significantly reduced biocontrol ability against cucumber damping-off caused by P. ultimum. Therefore, we investigated the influence of laboratory culturing media on biocontrol activity and physiological state of Burkholderia gladioli strain B543 by using long-term repeated culture on a certain medium. When isolate B543 were successionally cultured on King's B agar (KBA), tryptic soy agar, nutrient agar (NA), or soil extract agar more than 20 times, the isolate cultured on KBA or NA showed a significantly enhanced biocontrol efficacy and higher population density in the rhizosphere of cucumber compared to that of the others. However, the isolates cultured on KBA more than 20 times showed the lowest production of protease, siderophore, or antifungal substance(s), measured by skim milk agar, Chrome-Azurol-S agar, and potato dextrose agar amended with 10% of the culture filtrate, respectively. Our results suggest that adaptation to proper culturing medium can alter biocontrol ability and physiological state, and we must consider laboratory media in optimizing the use of biocontrol agents.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.105-113
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• 2019

Although siderophore compounds are mainly biosynthesized as a response to iron deficiency in the environment, they also bind with other metals. A few studies have been conducted on the impact of heavy metals on the siderophore-mediated iron uptake by microbiome. Here, we investigated siderophore production by a variety of rhizosphere fungi under different concentrations of $Zn^{2+}$ ion. These strains were specifically isolated from the rhizosphere of Panax ginseng (Korean ginseng). The siderophore production of isolated fungi was investigated with chrome azurol S (CAS) assay liquid media amended with different concentrations of $Zn^{2+}$ (50 to $250{\mu}g/ml$). The percentage of siderophore units was quantified using the ultra-violet (UV) irradiation method. The results indicated that high concentrations of $Zn^{2+}$ ion increase the production of siderophore in iron-limited cultures. Maximum siderophore production by the fungal strains was detected at $Zn^{2+}$ ion concentration of $150{\mu}g/ml$ except for Mortierella sp., which had the highest siderophore production at $200{\mu}g/ml$. One potent siderophore-producing strain (Penicillium sp. JJHO) was strongly influenced by the presence of $Zn^{2+}$ ions and showed high identity to P. commune (100% using 18S-rRNA sequencing). The purified siderophores of the Penicillium sp. JJHO strain were chemically identified using UV, Fourier-transform infrared spectroscopy (FTIR), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS) spectra.

• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.461-470
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• 1999

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrome azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-restricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.

• Journal of Microbiology and Biotechnology
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• v.23 no.5
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• pp.652-655
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• 2013

Diverse p-terphenyl compounds, named curtisians, have been isolated from the fungus Paxillus curtisii, and degradation of wood by this fungus is thought to be progressed by iron chelation of p-terphenyl curtisians. In this study, the iron chelation ability of p-terphenyls has been proved by chrome azurol S (CAS) assay, reducing power, and UV-visible spectroscopic analyses. The catechol moiety of p-terphenyl is an essential factor for the potent iron chelation ability, and thus deacylated curtisian with a tetrahydroxyl moiety in the central ring of p-terphenyl is more effective than acylated curtisians.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.415-421
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• 2003

A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.

• The Plant Pathology Journal
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• v.17 no.3
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• pp.174-179
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• 2001

The plant growth-promoting Pseudomonas strains, WR8-3 (Pseudomonas fluorescens), WR9-11 (Pseudomonas sp.) and WR9-16 (P.putida), which induced resistance systematically in watermelon to gummy stem rot were investigated on their induced systemic resistance(ISR)-related characteristics. The pyoverdine production was repressed in the standard succinate medium by increasing the concentration of $\textrm{FeCL}_3$. But the iron-binding ability on chrome azurol S agar media (CAS) was observed only in the strains, WR8-3 and WR9-16. When the two strains were mutated, the resulting iron-binding siderophore-negative mutants, WR8-3m and WR 9-16m, failed to promote the growth of watermelon and to induce resistance. The strains, WR8-3 and WR 9-16, slightly inhibited the growth of Didymella bryoniae at a low concentration of $\textrm{FeCL}_3$ on Kong's medium B, but not to exert control dffect. The strain WR9-11 showed antagonism in the concentration of $\textrm{FeCL}_3$ from 0 to $1,000\mu\textrm{M}$. When the crude lipoplysaccharide of each strain was treated in the rhizosphere of watermelon, mean lesion area was similar to that of the untreated control. The strains, WR9-11 and WR9-16 produced some level of hydrogen cyanide (HCN). Salicylic acid production was not detected in all of the strains.

• The Plant Pathology Journal
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• v.35 no.5
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• pp.425-436
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• 2019

Botrytis cinerea, a major phytopathogenic fungus, has been reported to infect more than 200 crop species worldwide, and it causes massive losses in yield. The aim of this study was to evaluate the inhibitory abilities and effects of Bacillus amyloliquefaciens RS-25, Bacillus licheniformis MG-4, Bacillus subtilis Z-14, and Bacillus subtilis Pnf-4 and their culture filtrates and extracts against the gray mold caused by B. cinerea on postharvest tomato, strawberry, and grapefruit. The results revealed that the cells of Z-14, culture filtrate of RS-25, and cells of Z-14 showed the strongest biocontrol activity against the gray mold on the strawberry, grape, and tomato fruit, respectively. All the strains produced volatile organic compounds (VOCs), and the VOCs of Pnf-4 displayed the highest inhibition values. Based on headspace solid-phase microextraction in combination with gas chromatography-mass spectrometry, esters accounted for the largest percentage of the VOCs produced by RS-25, MG-4, Z-14, and Pnf-4 (36.80%, 29.58%, 30.78%, and 36.26%, respectively). All the strains showed potent cellulase and protease activities, but no chitinase activity. RS-25, Z-14, and MG-4, but not Pnf-4, grew on chrome azurol S agar, and an orange halo was formed around the colonies. All the strains showed biofilm formation, fruit colonization, and lipopeptide production, which may be the main modes of action of the antagonists against B. cinerea on the fruit. This study provides the basis for developing natural biocontrol agents against the gray mold caused by B. cinerea on postharvest fruit.

• 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.

• Microbiology and Biotechnology Letters
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• v.38 no.3
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• pp.295-301
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• 2010

Sheath rot and dry rot disease caused by Pseudomonas marginalis and Fusarium oxysporum were serious problems in garlic farmland. In this study, total of 160 indigenous antagonistic bacteria were isolated from 16 farmlands in Yeongcheon, Korea. Among these, 15 strains were able to inhibited P. marginalis and F. oxysporum. The 16s rDNA genes of the selected 15 strains were amplified and sequenced. The strains has strong antagonistic ability against garlic pathogens was achieved Bacillus subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240. The selected 3 strains tested for investigation of antifungal mechanisms further analyses; 3 strains of these validated for production of siderophore, ${\beta}$-glucanase and chitinase using CAS (chrome azurol S) blue agar, CMC-congo red agar and DNS method. The 3 strains were able to utilized insoluble phosphate as dertermined by vanado-molybdate method. The 3 strains verified for production of auxin and gibberellic acid using Salkowski test and holdbrook test. Also, 3 strains showed stimulation germination, stem growth promoting activity on the in vivo test. The 3 strains were able to effectively suppress P. marginalis and F. oxysporum causing sheath rot and dry rot diseases on the in vivo pot test.