• The Plant Pathology Journal
• /
• v.23 no.3
• /
• pp.180-186
• /
• 2007

The biocontrol agent, Serratia plymuthica A21-4, has been developed for controlling pepper Phytophthora blight. Serratia plymuthica A21-4 strongly inhibits the mycelial growth, zoospore formation, and cyst germination of Phytophthora capsici in vitro. The application of a cell suspension of strain A21-4 to pepper plants in pot experiments and in greenhouse successfully controlled the disease. The bacteria produced a potent antifungal substance which was a key factor in the suppression of Phytophthora capsici. The most active chemical com-pound was isolated and purified by antifungal activity-guided fractionation. The chemical structure was identified as a chlorinated macrolide $(C_{23}H_{31}O_8Cl)$ by spectroscopic (UV, IR, MS, and NMR) data, and was named macrocyclic lactone A21-4. The active compound significantly inhibited the formation of zoosporangia and zoospore and germination of cyst of P. capsici at concentrations lower than $0.0625{\mu}g/ml$. The effective concentrations of the macrocyclic lactone A21-4 for $ED_{50}$ of mycelial growth inhibition were $0.25{\mu}g/ml,\;0.25{\mu}g/ml,\;0.30{\mu}g/ml \;and\;0.75{\mu}g/ml$ against P. capsici, Pythium ultimum, Sclerotinia sclerotiorum and Botrytis cinerea, respectively.

• The Plant Pathology Journal
• /
• v.28 no.2
• /
• pp.156-163
• /
• 2012

Stem rots caused by Rhizoctonia solani and Sclerotinia sclerotiorum have been known as devastating diseases in balloon flower plants. Antifungal activities of four fungicides, azoxystrobin, polyoxin B, trifloxystrobin and validamycin A were evaluated in vitro, showing effective suppression with mycelial growth of the fungal isolates on PDA media. Efficacies of the four fungicides were also demonstrated in stem tissues of balloon flower plants against R. solani and S. sclerotiorum. A commercially available Bacillus subtilis strain Y1336 was tested in terms of antagonistic biological control of stem rot disease of balloon flower plants. The bacterial strain revealed its antifungal activities against R. solani and S. sclerotiorum demonstrated by dual culture tests using paper discs and two plant pathogenic fungi on PDA media, as well as by plant inoculation assay, indicating that this antagonistic bacterial strain can be incorporated into disease management program for balloon flower stem rot diseases together with the four chemical fungicides.

• Mycobiology
• /
• v.37 no.2
• /
• pp.114-120
• /
• 2009

Streptomyces albidoflavus C247 was isolated from the soil of the Gyeongsan golf course in Korea. Physiological, biochemical and 16S rDNA gene sequence analysis strongly suggested that the isolate belonged to Streptomyces albidoflavus. Preliminary screening revealed that the isolate was active against fungi and bacteria. Self-directing optimization was employed to determine the best combination of parameters such as carbon and nitrogen source, pH and temperature. Nutritional and culture conditions for the production of antibiotics by this organism under shake-flask conditions were also optimized. Maltose (5%) and soytone (5%) were found to be the best carbon and nitrogen sources for the production of antibiotics by S. albidoflavus C247. Additionally, 62.89% mycelial growth inhibition was achieved when the organism was cultured at $30^{\circ}C$ and pH 6.5. Ethyl acetate (EtOAc) was the best extraction solvent for the isolation of the antibiotics, and 100 ${mu}$/ml of EtOAc extract was found to inhibit 60.27% of the mycelial growth of Rhizoctonia solani AG2-2(IV) when the poison plate diffusion method was conducted.

• The Korean Journal of Mycology
• /
• v.49 no.4
• /
• pp.441-454
• /
• 2021

Fungal diseases including anthracnose, stem rot, blight, wilting, and root rot of crops are caused by phytopathogens such as Colletotrichum species, Sclerotinia sclerotiorum, Phytophthora species, and Fusarium oxysporum and F. solani which threaten the production of chili pepper. In this study, to identify biological control agents (BCAs) of phytopathogenic fungi, potentially useful Bacillus species were isolated from the field soils. We screened out five Bacillus strains with antagonistic capacity that are efficiently inhibiting the growth of phytopathogenic fungi. Bacillus species were characterized by the production of extracellular enzymes, siderophores, and indole-3-acetic acid (IAA). Furthermore, the influence of bacterial strains on the plant growth promoting activity and seedling vigor index were assessed using Brassica juncea as a model plant. Inoculation with Bacillus subtilis SRCM 121379 significantly increased the length of B. juncea shoots and roots by 45.6% and 52.0%, respectively. Among the bacterial isolates, Bacillus subtilis SRCM 121379 showed the superior enzyme activities, antagonistic capacity and plant growth promoting effects. Based on the experimental results, Bacillus subtilis SRCM 121379 (GenBank accession no. NR027552) was finally selected as a BCA candidate.

• KSBB Journal
• /
• v.29 no.3
• /
• pp.131-138
• /
• 2014

In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

• Applied Biological Chemistry
• /
• v.34 no.2
• /
• pp.180-186
• /
• 1991

A water soluble antifungal antibiotic, Solumycin, was separated from the culture broth of Streptomyces sp. LAM-593, isolated from soil, by butanol extraction, alumina-, 1st and 2nd Sephadex LH-20 column chromatography. The substance was pale yellow crystal which gave a single spot at Rf value 0.24 with ethanol-ammonia water-water (8:1:1), 0.46 with butanol-ethanol-water (5:1:4), 0.84 with 50% methanol on silica gel TLC. It was dissolved well in water, methanol and acidic aq. butanol but not in ethanol, acetone, ethyl acetate, chloroform. acetic acid etc., and gave positive Fehling and Molish reaction. The UV spectrum in methanol showed absorption at 342, 361, 380, and 404 nm. The antibiotic was active against fungi such as Candide, Cryptococcus, Saccharomyces, Trichophyton and Trichosporon, but not to bacteria such as Bacillus, Escherichia and Staphylococcus.

• Journal of Microbiology and Biotechnology
• /
• v.4 no.4
• /
• pp.296-304
• /
• 1994

Antibiotic substances were produced by Bacillus subtilis YB-70, a potential biocontrol agent found to suppress root-rot of eggplant (Solanum melonggena L) caused by Fusarium solani, in a dextrose glutamate medium and isolated by isoelectric precipitation. Partial purification was performed by column chromatography on silica gel with two solvent systems: chloroform-methanol and methanol-chloroform-water as eluting solvents, This active fraction YBS-1 s contained antifungal activity were soluble in ethanol, methanol, and water, but were not soluble in other solvents including acetone, butanol, ethyl ether, dimethylformamide, propanol, and etc. High performance liquid chromatography and thin layer chromatographic separation of YBS-1s showed that they have been composed of three biological active bands that were named YBS-1A, -1B, and -1C. The substances were stable to heat and resistant to protease. YBS-1s were active against a wide range of plant pathogenic fungi but did not inhibit the growth of bacteria and yeasts. They were not only fungicidal but also fungistatic against chlamydospores of F. solani. The $ED_{50}$ values for the chlamydospore germination and the germ-tube growth of F. solani were $O.725\mu\textrm{m}/ml\;and\;O.562\mu\textrm{m}/ml$, respectively. Microscopic observations proved the substances restricted the growth of phytopathogenic fungus F. solani by spore burst followed by dissolving of its germ-tube, and caused abnormal hyphal swelling after application to chlamydospores or growing hyphae. Cultural filtrate of B; subtilis YB-70 also suppressed the development of root-rot of eggplant in pot tests.

• Journal of Ginseng Research
• /
• v.42 no.3
• /
• pp.304-311
• /
• 2018

Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.

• Mycobiology
• /
• v.43 no.3
• /
• pp.333-338
• /
• 2015

In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria.

• Journal of Life Science
• /
• v.9 no.1
• /
• pp.1-7
• /
• 1999

To isolate of antagonistic bacteria to Phytophthora capsici, which cause Phytophthora blight in red pepper, 237 isolates of Pseudomonas spp. and 260 isolates of Bacillus spp. were screened in selective media from rhizosphere soils of red pepper at Kyongsan, Kyongju, Yongchon and Euisung in Kyongbuk. Among total 497 isolates, 8 isolates of Pseudomonas spp and 4 isolates of Bacillus spp. inhibited the mycelial growth of Phytophthora capsici above 50$\%$ . These antagonistic bacteria showed more inhibitory effect on TSA (tryptic soy agar) than V-8 juice agar. Four isolates, P0704, P1201, B1101 and B1901, showing the most prominent antagonistic activity were selected and identified as P. cepacia (P0704, P1201), B. polymyxa (B1101) and B. subtilis (B1901), respectively. Cell free filtrates of these isolates were shown to inhibit zoosporangia germination and mycelial growth of p. capsici indicating that these isolates turned out to be bacteria producing antifungal substances. As a result of antagonistic test to Phytophthora blight in green house p. cepacia (P0704) showed the highest antagonistic effect with 46.7$\%$ and the rest of them were in the range of 13.4$\%$ to 26.7$\%$ .