• Korean Journal of Organic Agriculture
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• v.30 no.3
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• pp.409-422
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• 2022

Southern blight caused by Sclerotium rolfsii is a serious soilborne disease in economically important crops including pepper. In this study, we conducted a selection of antagonistic bacterial strains and organic materials to biologically control the disease. Out of 39 strains isolated from soils at Jinju in Korea, strain JJ2-01 showed the highest mycelial growth inhibition; garlic oil among various organic materials significantly reduced disease incidence and severity. When a combination of strain JJ2-01 and garlic oil, or each was drenched into the pepper plants, combined treatment and garlic oil significantly suppressed the disease development, however, acid phosphatase activity in garlic oil-treated plants decreased. In the case of combined treatment, the soil activities did not affect by treatment, while soil urease activity was significantly increased by the combined treatment. Therefore, given soil quality and health for sustainable agriculture, the combination of strain JJ2-01 and garlic acid was an effective application for environmental-friendly control of Southern blight in pepper plants.

• Research in Plant Disease
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• v.15 no.2
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• pp.88-93
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• 2009

In 2008, leaf rot and blight on pepper seeding ("Dokya-chungchung") occurred in a pepper farm at Hwaseong-si, Gyeonggi-do, Korea. The typical symptom is water-soaking and dark brown leaf blight at edges and tips of leaves. The fungal colonies isolated from infected tissues were pinkish at first and turned gradually to gray. Conidia were fusiform, non-septum, and $8.1-17.0{\times}2.0-3.8{\mu}m$ in size. Several specific PCR primers derived from the sequence of the internal transcribed spacer (ITS) region of the rDNA, such as CaINT, CgINT and CcINT were used for the identification of the fungal pathogen. The C. acutatum-specific primer CaINT was amplified single fragment of 496 bp that discriminated C. acutatum from the other species. The pathogenicity test was performed on seedlings and fruits of red pepper. On the basis of the morphological, molecular characteristics and pathogenicity test, we identified as Colletotrichum acutatum. This is the first report on leaf rot and blight on pepper seedling caused by C. acutatum in Korea.

• 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.

• Mycobiology
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• v.29 no.4
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• pp.218-223
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• 2001

A rapid radicle assay for prescreening antagonistic bacteria to Phytophthora capsic4 causal agent of Phytophthora blight of pepper was developed. Sixty-four bacterial strains with in vitro antifungal activity selected out of 1,400 strains isolated from soils of Ansung, Chunan, Koyang, and Paju, Korea in 1998 were used for development of the bioassay. Uniformly germinated pepper seeds dipped in bacterial cells for 3 hours were placed near the edges of growing mycelia of P. capsici on water agar containing 0.02% glucose. Five-week-old pepper plants(cv. Nockwang) were inoculated to compare with results of the radicle assay developed in this study. For plant inoculation, pepper seeds were sown in potting mixtures incorporated with the bacterial strains, then transplanted into steam-sterilized soils 3 weeks later. Plants were hole-inoculated with zoospores of P. capsici 2 weeks after transplanting. Disease incidence and severity were determined in radicle and plant assessments, respectively. In radicle assay, six strains, GK-B15, GK-B25, OA-B26, OA-B36, PK-B09, and VK-B14 consistently showed the significant(P=0.05) disease reduction against radicle infection by the fungus, four of which also did in plant assessments. Strains OA-B36 and GK-B15 consistently reduced the fungal infection in both the radicle assay and the plant assessment. Therefore, prescreening strains using the radicle assay developed in this study followed by plant assay could reduce time and labor, and improved the possibility of selecting antagonistic bacteria for control of Phytophthora blight of peppers.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.123-135
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• 2023

Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO₄ solution (untreated control). In diluted (nutrient-deficient) V₈ juice broth, the tested strain populations were maintained at >10⁸ cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 10⁶ cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.

• Research in Plant Disease
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• v.16 no.2
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• pp.170-175
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• 2010

A monoterpenoid benzylideneacetone (BZA) is a bacterial metabolite isolated from culture broth of an entomopathogenic bacterium, Xenorhabdus nematophila K1. It was tested in this study the control efficacy of the metabolite against two major fungal diseases occurring in red-pepper plants. BZA exhibited significant antifungal activities against Phytophthora capsici and Colletotrichum acutatum. Under natural light conditions, the antifungal activity of BZA was maintained for more than sixty days. The antifungal activity of BZA was not lost even in soil because the incidence of Phytophthora blight against red-pepper plants was significantly reduced when the suspensions of P. capsici were poured to the rhizosphere soils mixed with BZA. Application of the BZA suspension spray to the fruit surface infected with C. acutatum significantly suppressed the disease occurrence of anthracnose on the red-pepper plants. These results suggest that BZA can be used to develop a promising agrochemical to control phytophthora blight and anthracnose of redpepper plants.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1001-1010
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• 2005

Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

• Research in Plant Disease
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• v.12 no.1
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• pp.46-50
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• 2006

The microorganisms with the antifungal activity against Phytophthora capsici and Colletotrichum acutatum and the plant growth-promoting activity were screened from a forest and natural fields of Gajang-Dong, Sangju-city. One of the isolates, Streptomyces griseofuscus 200401, was selected as a good plant growth-promoting strain in this study. In greenhouse test, the number of leaf, fresh weight, and dry weight of pepper plants, that were grown with treatment of culture suspension or powder containing S. griseofuscus 200401, were higher than those without the bacterial cells. Cultivation of S. griseofuscus 200401 strain for 7 days in a nutrient rich medium produced ammonium chloride up to 0.13 ${\mu}g/ml$ in the culture solution of S. griseofuscus. Treatment of the selected strain significantly reduced the severity of the late blight of pepper plants to show the equivalent disease control activity to chemical fungicide. This study suggests that S. griseofuscus 200401 strain could be a potential biological agent with the biocontrol activity and the plant growth-promoting activity.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.340-345
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• 2010

Two similar microbial fungicides (termed as MA and MB) developed in a Korean biopesticide company were analyzed and compared each other in their biocontrol activities against the phytophthora blight of chili pepper caused by Phytophthora capsici. MA and MB contained the microbe Paenibacillus polymyxa and Bacillus subtilis, respectively, with concentrations over those posted on the microbial products. In comparison of the isolated microbes (termed as MAP from MA and MBB from MB) in the antagonistic activities against P. capsici was effective, prominently against zoospore germination, while MBB only significantly inhibited the mycelia growth of the pathogen. Some effectiveness of MAP and MBB was noted in the inhibition of zoosporangium formation and zoospore release from zoosporangia; however, no such large difference between MAP and MBB was noted. In a pot experiment, MA reduced the severity of the phytophthora blight more than MB, suggesting that the disease control efficacy would be more attributable to the inhibition of zoospore germination than mycelia growth of P. capsici. These results also suggest that the similar microbes MA and MB targeting different points in the life cycle of the pathogen differ in the disease control efficacies. Therefore, to develop microbial fungicides it is required to examine the targeting points in the pathogen's life cycle as well as the action mode of antagonistic microorganisms.

• The Plant Pathology Journal
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• v.15 no.4
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• pp.217-222
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• 1999

Treatment with antagonistic rhizobactera Burkholderia cepacia strain N9523 or an inducer of resistance DL-$\beta$-amino-n-butyric acid (BABA) effectively inhibited Phytophthora capsici infection on pepper plants in artificially infested pots. Treatment with BABA alone at $1,000\mu\textrm{g}$/ml or together with B. cepacia in combination induced a strong protection from the Phytophthora disease in the greenhouse. In artificially infested field, protection of pepper plants against the Phytophthora epidemic by BABA treatment was maintained at a considerable level. In contrast, soil drench with the antagonist B. cepacia alone, or in combination with BABA did not suppress the Phytophthora epidemic in the field. Mortality of pepper plants caused by P. capsici infection was significantly reduced by treatment with the antagonist Pseudomonas aeruginosa strain 950923-29 and BABA (12-29% plants diseased) relative to the untreated control (41-91% plants diseased) in the naturally infested field. Treatment with the antagonist Ps. aeruginosa strain 950923-29 and BABA also resulted in high levels of protection against Phytophthora blight in pepper plants. In the plastic filmhouse test, the average percentage of plants diseased was significantly low relative to the naturally infested field. Treatment with the antagonist Ps. aeruginosa strain 950923-29 and BABA in combination was most effective in suppressing the Phytophthora disease in field and plastic filmhouse.