• The Plant Pathology Journal
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• v.38 no.1
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• pp.25-32
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• 2022

Plant pathogens pose major threats on agriculture and horticulture, causing significant economic loss worldwide. Due to the continuous and excessive use of synthetic pesticides, emergence of pesticide resistant pathogens has become more frequent. Thus, there is a growing needs for environmentally-friendly and selective antimicrobial agents with a novel mode of action, which may be used in combination with conventional pesticides to delay development of pesticide resistance. In this study, we evaluated the potentials of lichen substances as novel biopesticides against eight bacterial and twelve fungal plant pathogens that have historically caused significant phytopathological problems in South Korea. Eight lichen substances of diverse chemical origins were extracted from axenic culture or dried specimen, and further purified for comparative analysis of their antimicrobial properties. Usnic acid and vulpinic acid exhibited strong antibacterial activities against Clavibacter michiganensis subsp. michiganensis. In addition, usnic acid and vulpinic acid were highly effective in the growth inhibition of fungal pathogens, such as Diaporthe eres, D. actinidiae, and Sclerotinia sclerotiorum. Intriguingly, the growth of Rhizoctonia solani was specifically inhibited by lecanoric acid, indicating that lichen substances exhibit some degrees of selectivity to plant pathogens. These results suggested that lichen substance can be used as a selective biopesticide for controlling plant disease of agricultural and horticultural significance, minimizing possible emergence of pesticide resistant pathogens in fields.

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.526-534
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• 2019

Chlorella gloriosa (Chlorellaceae, Trebouxiophyceae) was isolated from seawater off the coast of the Dokdo Islands in Korea. An axenic culture was established using the streak-plate method on f/2 agar media supplemented with antibiotics, allowing identification of the isolate by morphological, molecular, and physiological analyses. The morphological characteristics observed by light and electron microscopy revealed typical morphologies of C. gloriosa species. The molecular phylogenetic inference drawn from the small-subunit 18S rRNA sequence verified that the microalgal strain belongs to C. gloriosa. Additionally, gas chromatography-mass spectrometry analysis showed that the isolate was rich in nutritionally important omega-3 and -6 polyunsaturated fatty acids and high-performance liquid chromatography analysis revealed that the high-value antioxidants lutein and violaxanthin were biosynthesized as accessory pigments by this microalga, with arabinose, galactose, and glucose as the major monosaccharides. Therefore, in this study, a Korean marine C. gloriosa species was discovered, characterized, and described, and subsequently added to the national culture collection.

• Korean Journal of Plant Resources
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• v.32 no.6
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• pp.730-734
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• 2019

Cryopreservation is one of the ideal and suitable methods for long-term storage of plant germplasm. The plant contaminated with diseases and pathogens are decreased the multiplication rate, survival rate and high quality of plants after cryopreservation. The aim of this work was to improve a micropropagation method for lily in Korea, which is indigenous plant. In the last process of rinsing scales after surface-sterilization, we tried to control the diseases and pathogens lived within the tissue by rinsing in 0.03% sodium hypochlorite (NaClO) instead of sterile distilled water. Bulb scales of Lilium were cultured in vitro on MS medium supplemented with Plant Preservative Mixture (PPM). The results showed that L. tsingtauense accessions were observed ranged from 53.9 to 100% with a mean value of 76.8% and L. hansonii accessions were checked from 84.5 to 85.5% with a mean of 85% survival rate. The newly small bulb formed from bulb-scales was transferred to MS medium. We checked the presence of microorganisms and survival rate after 3 weeks in culture after examination of bacterial incidences. The results indicated that the non-contamination rate were shown ranged from 75.0 to 94.1% with mean value of 83.2% in L. tsingtauense species, and that L. hansonii were observed 85.1 to 91.7% with mean value of 88.4%. This study will provide a valuable basis for establishment of effective axenic cultures for in vitro micropropagation of Korean native lily species.

• The Korean Journal of Mycology
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• v.42 no.4
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• pp.312-321
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• 2014

Tricholoma bakamatsutake is one of the edible ectomycorrhizal mushrooms as an allied species of Tricholoma matsutake. This is the first report on physical characteristics of T. bakamatsutake strains collected from Quercus mongolica forests in Korea. The pure cultures of these strains were isolated from the tissues of fruit bodies and the culture characteristics were investigated under different conditions (media, temperatures, nitrogen sources). Most strains showed the highest mycelial growth on potato dextrose agar (PDA) at 20 or $25^{\circ}C$. Two strains of T. bakamatsutake preferred the ammonium-form rather than the nitrate-form as an inorganic nitrogen source. T. bakamatsutake showed significantly slower mycelial growth when compared with T. matsutake from a Korean forest, although the optimum culture conditions for the two allied species were similar. We also tested the ability to form mycorrhizae as well as cellulase activity of T. bakamatsutake. All strains showed cellulase activity on a carboxymethylcellulose (CMC) agar plate. The mycorrhizae on axenic Pinus densiflora seedlings were formed by two strains of T. bakamatsutake after 3 or 8 months of inoculation. P. densiflora seedlings inoculated with T. bakamatsutake had a much higher biomass than un-inoculated seedlings.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.1
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• pp.71-76
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• 1998

To know the antibiotic specificity of a Dinoflagellate, Cochlodinium polykrikoides, we investigated the survival time of C. polykrikoides against several concentrations of antibiotics and judged the selective specificity of antibiotics based on the $LT_50$ ($50\%$ of lethal time). The result showed that C. polykrikoides was sensitive to tetracycline and chloramphenicol, and resistant to polymixin-B, ampicillin, penicillin-G, dihydrostreptomycin, and neomycin. In the case of sensitive antibiotics to C. polykrikoides, tetracycline and chloramphenicol, the safety concentrations of both antibiotics were determined and the antibiotic specificity based or the plotted survival curve was analyzed. Before antibiotic treatment, we tested the antibiotic susceptibility of the contaminated bacterial population in tile culture of C. polykrikoides, and decided the proper kinds of antibiotics and concentrations before percoll-centrifugation. By percoll-centrifugation, we reduced bacteria, removed fungi, collected the algal pellet, and made axonic culture by antibiotic cascade procedure based on the result of antibiotic susceptibility test. We observed that axonic C. polykrikoides culture entered the logarthmic phase of growth when cell density was over 740 cells/ml and propagated to 5,800 cells/ml maximally. Divisions per day, k value of C. polykrikoides represented a good index for growth at the low density of cells. There was a highest k value shift before reaching to the logarithmic phase. We suggested that the preceeding highest k value shift stage is a good indicator for accurate broadcasting for red. tide blooming in the field, and the stage is also a good time for controlling red tide blooming in the filed, either.

• Proceedings of the Microbiological Society of Korea Conference
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• 2001.11a
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• pp.46-53
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• 2001

A commensal thermophile, Symbiobacterium toebii, isolated from hay compost (toebii) in Korea commensally interacted with a thermophilic Geobacillus toebii sp. nov., which was a new species within the genus Geobacillus on the basis of the phenotypic traits and molecular systematic data. S. toebii required the crude extracts and/or culture supernatant of the Geobacillus toebii for axenic growth and could grow on the temperature between 45 and $70^{\circ}C$ (optimum: $60^{\circ}C$; 2.4 h doubling time) and pH 6.0 and 9.0 (optimum: pH 7.5). The G+C content of the genomic DNA was $65 mol\%$, and the major quinones were MK-6 and MK-7. A phylogenetic analysis of its 16S rDNA sequence indicated that Symbiobacterium toebii was closely related with solely reported Symbiobacterium thermophilum. The presence of the commensal thermophile 16S rDNA and accumulation of indole in all the enriched cultures indicate that Symbiobacterium toebii is widely distributed in the various soils. The genome of S. toebii constituted a circular chromosome of 3,280,275 base pairs and there was not an extra-chromosomal element (ECE). It contained about 4,107 predicted coding sequences. Of these protein coding genes, about $45.6\%$ was encoded well-known proteins and annotated the functional assignment of 1,874 open reading frames (ORFs), and the rest predicted to have unknown functions. The genes encoding thermostable tyrosine phenol-lyase and tryptophan indole-lyase were cloned from the genomic DNA of S. toebii and the enzymatic production of L-tyrosine and L-tryptophan was carried out with two thermostable enzymes overexpressed in recombinant E. coli.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.4
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• pp.266-287
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• 2004

The non infecting, plant associated bacteria have attracted increased attention for stimulating plant growth and as environmental friendly plant protecting agents. Pink-pigmented facultatively methylotrophic bacteria (PPFMs), classified as Methylobacterium spp., are persistent colonizers of plant leaf surfaces. As the leaves of most or all plants harbor PPFMs that utilize leaf methanol as their sole source of carbon and energy, which is a specific attribute of the genus Methylobacterium. Although they are not well known, these bacteria are co-evolved, interacting partners in plant metabolism. This claim is supported, for example, by the following observations: (1) PPFMs are seed-transmitted, (2) PPFMs are frequently found in putatively axenic cell cultures, (3) Low numbers of seed-borne PPFMs correlate with low germinability, (4) Plants with reduced numbers of PPFM show elevated shoot/root ratios, (5) Foliar application of PPFMs to soybean during pod fill enhances seed set and yield, (6) Liverwort tissue in culture requires PPFM-produced vitamin B12 for growth, (7) treated plants to suppress or decrease disease incidence of sheath blight caused by Rhizoctonia solani in rice, and (8) the PPFM inoculation induced number of stomata, chlorophyll concentration and malic acid content, they led to increased photosynthetic activity. Methylobacterium spp. are bacterial symbionts of plants, shown previously to participate in plant metabolism by consuming plant waste products and producing metabolites useful to the plant. There are reports that inform about the beneficial interactions between this group of bacteria and plants. Screening of such kind of bacteria having immense plant growth promoting activities like nitrogen fixation, phytohormone production, alleviating water stress to the plants can be successfully isolated and characterized and integration of such kind of organism in crop production will lead to increased productivity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.3
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• pp.364-372
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• 1995

On the outbreak of paralytic shellfish poisoning in April 1993 in most of shellfish harvesting areas in Jinhae Bay, Korea, to clarify the toxin production of causative organism Alexandrium species, 19 axenic clonal isolates established from the benthic resting cysts in three different stations of those culture grounds were subjected to PSP toxin analysis by HPLC. Individual toxin content per cell was highly variable among the strains isolated from a sampling area and originated from an individual cyst. Average toxin contents in those areas revealed higher values of 54-70 fmol/cell. Toxin profiles included C1/C2(epiGTX8/GTX8), GTX1/GTX4 and neoSTX as the major components, and GTX2/GTX3, GTX5, C4, dcSTX and STX as the minor or sporadic ones. neoSTX on the dominant toxins showed not only most diverse compositional changes comprising $5-54 mol\%$ ranges but also no detection on the half of the strains examined, which were implicated in arising of heterogeneity with a genetic trait within a geographical region. When average toxin composition was compared, carbamate toxins comprised large proportions of $57\%,\;54\%\;and\;67\%$ as total toxin in St. 1, St. 2 and St. 4, respectively. These results suggested that an extensive paralytic shellfish toxification in Jinhae Bay could be largely due to the production of highly potent carbamate toxins in the causative dinoflagellate Alexandrium species.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.41-45
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• 2002

An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.

• Korean Journal of Ecology and Environment
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• v.34 no.1 s.93
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• pp.20-29
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• 2001

The amount, chemical composition and optical property of extracellular dissolved organic carbon (EOC) by phytoplankton were examined using axenic cultures of Microcystis aeruginosa, Anabaena flos-aquae, and Oscillatoria agardhii. The extracellular organic matter was categorized into five fractions (hydrophobic acids; AHSs, hydrophobic neutrals; HoNs, hydrophilic acids; HiAs, hydrophilic bases; HiBs, and hydrophilic neutrals; HiNs) using three adsorbent resins(XAD-8, cation, and anion). The release pattern and chemical composition of EOC varied with algal species and their growth phases. Percentage of extracellular release increased with age in all cultures. HiAs were the dominant component of EOC in all cultures, whereas the proportion of HiAs decreased with age in all cultures. In contrast, the proportions of HiBs and HiNs increased as cultures aged. In particular, the HiN fraction increased from 0% to 44% of EOC in M. aeruginosa and from 3.0% to 28% in A. flos-aquae, respectively. The proportion of AHSs was higher in the cultures of A. flos-aquae(7.5${\sim}$16%) and O. agardhii (8.7${\sim}$16%) than M. aeruginosa(0.2${\sim}$2.5%). The proportions of AHSs increased with culture age in M. aeruginosa and O. agardhii, but decreased in A. flos-aquae. The specific UV absorbance also varied among species; 1.9 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for M. aeruginosa, 3.7 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for A. flos-aquae, and 13.0 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L^{-1}$ for O. agardhii. The results of this study indicates that DOC excreted by three blue-green algae differed with species and the growth phase.