• 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.24 no.2 s.77
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• pp.111-126
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• 1996

Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.43-50
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• 2017

A xylan-degrading bacterium (strain WJ-1) was isolated from soil collected from Jeju Island, Republic of Korea. Strain WJ-1 was characterized as a gram-positive, aerobic, and spore-forming bacterium. The predominant fatty acid in this bacterium was anteiso-$C_{15:0}$ (42.99%). A similarity search based on 16S rRNA gene sequences suggested that the strain belonged to the genus Streptomyces. Further, strain WJ-1 shared the highest sequence similarity with the type strains Streptomyces spinoveruucosus NBRC 14228, S. minutiscleroticus NBRC 13000, and S. glaucescens NBRC 12774. Together, they formed a coherent cluster in a phylogenetic tree based on the neighbor-joining algorithm. The DNA G+C content of strain WJ-1 was 74.7 mol%. The level of DNA-DNA relatedness between strain WJ-1 and the closest related species S. glaucescens NBRC 12774 was 85.7%. DNA-DNA hybridization, 16S rRNA gene sequence similarity, and the phenotypic and chemotaxonomic characteristics suggest that strain WJ-1 constitutes a novel subspecies of S. glaucescens. Thus, the strain was designated as S. glaucescens subsp. WJ-1 (Korean Agricultural Culture Collection [KACC] accession number 92086). Additionally, strain WJ-1 secreted thermostable endo-type xylanases that converted xylan to xylooligosaccharides such as xylotriose and xylotetraose. The enzymes exhibited optimal activity at pH 7.0 and $55^{\circ}C$.

• The Korean Journal of Mycology
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• v.44 no.3
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• pp.201-205
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• 2016

In August 2015, leaf spot symptoms were observed on Korean gondre thistle (Cirsium setidens) in Youngwol, Korea. During the early stage, the symptoms appeared as one or more small gray-brown to brown spots on plant leaves. The spots showed extensive enlargement over time and eventually became large dark brown to black lesions on the whole leaf. Stemphylium species were consistently isolated from affected leaves. All isolates were identified as S. lycopersici, S. solani, or S. xanthosomatis based on morphological and cultural characteristics. The isolates were confirmed as S. lycopersici based on a multilocus sequence analysis using the ribosomal internal transcribed spacer (ITS) region, elongation factor 1, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and the noncoding region between the vacuolar membrane ATPase catalytic subunit A gene and a gene involved in vacuolar biogenesis. Pathogenicity was tested by spore suspension inoculation on wounded or unwounded gondre leaves. The lesions were observed on inoculated leaves within 3 days after inoculation, regardless of wound. To our knowledge, this is the first report of the leaf spot on gondre thistle caused by S. lycopersici in Korea or elsewhere.

• 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.

• Journal of agriculture & life science
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• v.46 no.3
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• pp.37-45
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• 2012

To select bacterial strains with antifungal activity against an anthracnose fungal disease causing damage severely on hot pepper, previous isolates obtained from plant root samples were screened. Among 457 isolates, IA70-5 isolate was finally selected and identified as Streptomyces padanus based on 16S rDNA sequence analysis. Strain IA70-5 is non-pigmenteous, non-mobile, and filamentous. S. padanus IA70-5 inhibited effectively the mycelium growth, spore germination, and appressorium formation of Colletotrichum acutatum in vitro. The results of this study demonstrated that IA70-5 strain, especially applied on fruit of hot pepper, decreased disease incidence 90% for pre-inoculation before pathogen treatment. Taken together, S. padanus IA70-5 strain is a promising biological control agent to control of a major fungal pepper disease, anthracnose.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.86-93
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• 2022

The ecotoxicity test method using Undaria pinnatifida spore is challenging to use throughout the year. Since U. pinnatifida female gametophytes can be cultured in the laboratory, they can be used for ecotoxicity testing at any time. Changes in female gametophyte survival rate and relative growth rate in U. pinnatifida exposed to various environmental conditions were analyzed. The female gametophyte of U. pinnatifida was exposed to salinity (5~40 psu), temperature (5~30℃), pH (4~10), and light intensity (0~120 μmol photon m^-2 s^-1). Based on the highest average value, the survival rate of female gametophyte was highest at a temperature of 20℃, salinity 27.5 psu, pH 8, and light intensity 30 μmol photon m^-2 s^-1. And the relative growth rate was highest at a temperature of 15℃, salinity 35 psu, pH 9, and light intensity of 60 μmol photon m^-2 s^-1. As a result of this study, the method using the optimal conditions for the survival rate and relative growth rate is expected to be a practical test method that can complement the current method.

• Journal of the Korea institute for structural maintenance and inspection
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• v.27 no.6
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• pp.138-143
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• 2023

In this study, the crack healing performance of each crack healing agent manufacturing method was analyzed by adding crack healing agents in the form of alginate gel and spore suspension inoculated with endospores of calcium carbonate-forming bacteria to mortar. In addition, by applying it to an full-scale structure in the form of a box-type culvert, we attempted to create an environment in which the developed crack healing agent can be applied not only to a laboratory environment but also to an actual field. The crack healing agent using the dry heat drying method showed crack healing performance, but in the case of the freeze drying method, many spores were killed by freeze hardening and therefore the crack healing performance was lost. As a result of SEM and XRD pattern analysis of the presumed crack healing material extracted from the crack of a full-scale structure, it was found to be calcite, one of the calcium carbonate crystals produced by microorganisms applied to the crack healing agent. In conclusion, it was found that the crack healing by microorganisms can be implemented in a real structure.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.53-53
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• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

• Journal of Nutrition and Health
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• v.18 no.4
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• pp.259-271
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• 1985

The purpose of this study was to develop supplementary foods for infants and young children in order to improve their nutritional status. Three formulas composed of rice, soybeans, fish, dry skim milk and sesame in varying proportions were studied. The three formulas, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$, were consisted of Rice(R), Soybean$(S_{1})$, Sesame$(S_{2})$ (60 : 35 : 5) , Rice, Fish(F), Soybean, Sesame (60 : 10 : 25 : 5) , and Rice, Dry Skin Milk (M), Soybean, Sesame (60 : 10 : 25 : 5), respectively. A proximate analysis and amino acid determination were made on the developed formulas. In the animal assay, growth rate, PER and FER were evaluated and biochemical analyses were also carried out. A storage test and the cost evaluation were also conducted. The summarized results are as follows : 1) The proximate composition of the three formulas were 7.3-7.4% of moisture, 15.9-21.5% of crude protein, 7.8-9.6% of crude fat and 2.5-2.8% ash. 2) The result of amino acid analysis showed that the 1st limiting amino acids of $RS_{1}S_{2}$ and $RFS_{1}S_{2}$ were lysine (amino acid score, 76.6) and threonine (amino acid score, 93.3), and that of $RMS_{1}S_{2}$ and the commercially prepared formula were sulfur containing amino acids (amino acid score, 82.0 and 54.4). When the contents of the amino acids of the three formulas were compared with mother's milk and cow's milk, the balance of the amino acid of each formula was superior to mother's milk but inferior to cow's milk. 3) In the animal assay, the growth rate of all groups increased gradually during the experimental period. 4) The C- PER, which was corrected on the basis of the casein PER of 2.5 was 2.99, 3.38 and 3.10 in the $RS_{1}S_{2}$, $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ respectively. The C- PER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were Significantly (P<.05) higher than that of the casein. 5) The FER of the casein, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$ were 0.37, 0.39, 0.43 and 0.39, respectively. The FER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were also significantly (P<.05) higher than that of the casein. 6) The concentrations of hematocrit, hemoglobin, total protein and albumin in the serum of the rats of all groups were not significantly different among groups. 7) The storage stability test showed that the total plate count (TPC), the coliforms count and the bacterial spore count in the ingredients were quiet low. However, after 30 and 60 days storage, the count in $RFS_{1}S_{2}$ increased and were higher at room temperature than refrigerated temperature. 8) In the cost evaluation, the cost of the developed formulas was \1,826-2,626 / kg. This was less than that of the commercially prepared formula (\3,300-4,073 / kg) and that of the imported formula (\4,250-8,720 / kg).