• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.194-199
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• 2005

In oder to select an antifungal substance-producing antagonistic bacterium against Fusarium oxysporum casuing fusarium wilt on tomato, strain BK4 was isolated from local soil of Gyeoungbuk and was identified as Bacillus thuringiensis by 16s rDNA analysis, biochemical test, and Mcirolog TM 3.0 System. The antibiotic of B. thuringiensis BK 4 was highly produced at $30^{\circ}C$ in nutrient broth (pH 9.0). The crude antibiotic was even stable at $121^{\circ}C$ and more stable at slight alkalic condition than acid condition. It was also remained $50{\%}$ activity at pH 3.0. B. thuringiensis BK4 showed the inhibition of spore germination and the biocontrol ability against F. oxysporum causing fusarium wilt of tomato in vivo test. According to these results, B. thuringiensis BK4 was enough to use with a microbial agent for biocontrol against fusarium wilt.

• The Korean Journal of Mycology
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• v.37 no.1
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• pp.96-103
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• 2009

For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.

• The Korean Journal of Mycology
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• v.39 no.2
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• pp.91-98
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• 2011

Oudemansiella mucida, an edible and medicinal mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to produce antifungal substances to inhibit the mycelial growth and spore germination of the plant pathogenic fungi. To produce good amount of antifungal substances from culture media, the optimal culture conditions of O. mucida were investigated. The most favorable conditions for the mycelial growth were $25^{\circ}C$ and pH 5 in potato dextrose agar. The most favorable carbon and nitrogen sources promoting mycelial growth were maltose and calcium nitrate, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplemented to the basal medium as a carbon source. The optimal mycelial growth of O. mucida was found in the Hennerberg medium. The crude extract from submerged culture of potato dextrose broth exhibited inhibition of mycelial growth of Colletotrichum acutatum, Botrytis cinerea and Pyricularia oryzae but, fungicidal activity is not good enough to compared with commercially available fungicides tested. Therefore, the antifungal substances extracted from submerged culture of O. mucida might have a potential to be used for biocontrol agent of fungal diseases of plants.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.15-15
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• 2014

Fusarium graminearum (teleomorph Gibberella zeae) is an important plant pathogen that causes head blight of major cereal crops such as wheat, barley, and rice, as well as causing ear and stalk rot on maize worldwide. Plant diseases caused by this fungus lead to severe yield losses and accumulation of harmful mycotoxins in infected cereals [1]. Fungi utilize spore production as a mean to rapidly avoid unfavorable environmental conditions and to amplify their population. Spores are produced sexually and asexually and their production is precisely controlled. Upstream developmental activators consist of fluffy genes have been known to orchestrate early induction of condiogenesis in a model filamentous fungus Aspergillus nidulans. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we characterized functions of the F. graminearum fluffy gene homologs [2]. We found that FlbD is conserved regulatory function for conidiogenesis in both A. nidulans and F. graminearum among five fluffy gene homologs. flbD deletion abolished conidia and perithecia production, suggesting that FlbD have global roles in hyphal differentiation processes in F. graminearum. We further identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum [3]. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. F. graminearum ortholog of Aspergillus nidulans wetA has been shown to be involved in conidiogenesis and conidium maturation [4]. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidia dormancy by suppressing microcycle conidiation in F. graminearum. In A. nidulans, FlbB physically interacts with FlbD and FlbE, and the resulting FlbB/FlbE and FlbB/FlbD complexes induce the expression of flbD and brlA, respectively. BrlA is an activator of the AbaA-WetA pathway. AbaA and WetA are required for phialide formation and conidia maturation, respectively [5]. In F. graminearum, the AbaA-WetA pathway is similar to that of A. nidulans, except a brlA ortholog does not exist. Amongst the fluffy genes, only fgflbD has a conserved role for regulation of the AbaA-WetA pathway.

• The Korean Journal of Pesticide Science
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• v.12 no.2
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• pp.168-176
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• 2008

Recently damage of rice bakanae disease disseminated by infected seeds increased in paddy field in Korea. For controlling rice bakanae disease, the efficacy of 17 fungicides was assessed by 5 kinds of bioassay, spore germination test (SGT), mycelial growth test, detection test on Komada's medium (KDT), pouch test (PT) and greenhouse test (GT). Among ergosterol biosynthesis inhibiting fungicides, prochloraz showed a high controlling activity in all the assay systems while the others showed very low activity except for $500\;{\mu}g/ml$ of hexaconazole in GT and $500\;{\mu}g/ml$ of triflumizole in KDT. Although benomyl and the mixture of benomyl and thiram showed a good activity at 100 and $500\;{\mu}g/ml$ in SGT and PT, respectively, in GT they did a middle activity. Trifloxystrobin and kresoxim-methyl included in strobilurins showed a good activity even at $20\;{\mu}g/ml$ in KDT as well as a middle activity in SGT. Also a high activity not only at $10\;{\mu}g/ml$ in SGT but also at $100\;{\mu}g/ml$ KDT was detected in thiram. The activity of fludioxonil was confirmed in SGT, KDT and PT. Based on these results, it is very important to determine a bioassay system, because the fungicidal activity against rice bakanae disease was fluctuated depending on a assay systems as well as the mechanism of fungicide.

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.128-134
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• 2007

Previously, we isolated plant growth promoting rhizobacterium (PGPR) Bacillus licheniformis K11 which could produce auxin, cellulase and siderophore. The siderophore of B. licheniformis K11 $(siderophore_{K11})$ was determined to be a catechol type siderophore which is produced generally by Bacillus spp. B. licheniformis K11 could produce the siderophore most highly after 96 h of incubation under nutrient broth at $20^{\circ}C$ with initial pH 9.0. For the production of the $siderophore_{K11}$, trehalose and $NH_4Cl$ were the best carbon and nitrogen sources in Davis minimal medium, respectively. The $siderophore_{K11}$ was Produced in M9 medium (pH 9.0) after 4 days at $20^{\circ}C$, and purified from culture broth of B. licheniformis K11 by using Amberlite XAD-2, Sephadex LH-20 column chromatography, and reversed-phase HPLC. The $siderophore_{K11}$ had the biocontrol activity against spore germination of P. capsici and F. oxysporum on potato dextrose agar (PDA). The results indicate that the $siderophore_{K11}$ is an antifungal mechanism of B. licheniformis K11 against phytopathogenic fungi.

• The Korean Journal of Mycology
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• v.22 no.1
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• pp.107-116
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• 1994

Interorder somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus in the order Agaricales and Elfvingia applanata in the order Aphyllophorales. The fusants were classified into stable heterokaryons and spontaneously segregated heterokaryons. Hyphae of all fusion products except two strains did not form clamp connections. Out of them, two clamped and three clampless fusants produced mature fruiting bodies by light-dark cycle on sawdust rice bran medium. All of these basidiocarps had clamp connections. Three fusants were analysed with the distribution of progenies and segregation of genetic characters by random spore analyses. The genetic markers were shown to segregate and recombine in the first generation of monospores isolated from basidiocarps. Phenotypes of a large number of auxotrophic progenies were not detected in the two clamped fusants. The aberration ratio of segregants indicated the gene interaction resulting from different genome structure between distantly related species. The polymerase chain reaction (PCR) was adopted for the detection of somatic hybrids nuclear DNA. Four fusants showed a positive results in three kinds of primers. The prominent reaction products are represented by new bands in primer # 87 and # 125. Out of four fusants, two somatic hybrids had non-parental mtDNA patterns when digested with EcoR1 and HindIII. Comparison of somatic hybrids, tissue culture isolates(TC) and multispore germination isolates(MS) were made using esterase isozyme analysis. It is apparent that somatic hybrids had a minor banding patterns which are quite different from those of parents.

• The Korean Journal of Pesticide Science
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• v.19 no.1
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• pp.47-53
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• 2015

Various diseases occur in fruits, leave and roots during strawberry cultivation and cause severe economical damage and huge amount of chemical fungicide use. Recently, as consumers' interest in safety of foods and organic agriculture produces have increased, control measures using alternatives for chemical fungicides have been newly developed in various ways. This study was conducted to test antifungal activity and control effect of sodium dichloroisocyanurate (NaDCC), using as disinfectants, against major disease pathogens of strawberry, Fusarium oxysporum (Fusarium wilt), Colletotrichum gloeosporioides (Anthracnose) and Phytophthora sp. (Phytophthora blight), and Xanthomonas fragariae (bacterial angular leaf spot) and evaluate availability as environment-friendly materials. When NaDCC was treated at the concentration range of 150 to 300 ppm, it suppressed significantly hyphal growth and reduced spore germination by more than 28%. In field condition, NaDCC showed excellent control effect (control value: 50%) against the bacterial angular leaf spot disease. Based on above-described results, we suggested that NaDCC can be used as alternative candidates to chemical pesticide alternatives of for controlling strawberry diseases.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.313-320
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• 2010

Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.855-868
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• 2014

Synthetic agro-chemicals have been widely used to control diseases on paprika but these days negative attention has been increasing to use of them because of several adverse effects. This research was conducted to isolate and to characterize the antagonistic microorganism to control major paprika diseases, gray mold rot, fruit and stem rot, phytophthora blight, sclerotium rot, and wilt disease. Analysis of the fatty acid and analysis of the 16S rDNA gene sequence revealed that YKJ1 isolated in this research belongs to a group of Serratia marcescens. Specially, 16S rDNA gene sequence of YKJ1 showed 99% of sequence similarity with S. marcescens. Observation through the optical microscope revealed that YKJ1 suppressed the spore germination and the hyphal growth of pathogens. YKJ1 treatment on pathogens induced marked morphological changes like hyphal swelling and degradation of cell wall. In the case of phytophthora blight, the zoosporangium formation was restrained. S. marcescens found in this study call as S. marcescens-YKJ1 and it may be valuable as one of biological control agents against major diseases of paprika in the future even though it is require to be tested with more study on field test.