• Research in Plant Disease
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• v.17 no.2
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• pp.121-128
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• 2011

Bacteriolytic myxobacteria have been known to secrete various antifungal metabolites against several soilborne phytopathogens including Phytophthora. Among the three isolates of Myxococcus spp., KYC 1126 and KYC 1136 perfectly inhibited the mycelial growth of Phytophtora capsici in vitro. In order to show the biocontrol activity on Phytophthora blight of hot pepper, we tried to find the best way of application of a myxobacterial isolate. Although KYC 1126 fruiting body was easily grown on the colony of Escherichia coli as a nutrient source, it did not control the disease when it was pre-applied in soil. Before the bioassay of a liquid culture filtrate of KYC 1126 was conducted, its antifungal activity was confirmed on the seedlings applying with the mixture of the pathogen's zoospore suspension and KYC 1126 filtrate. On greenhouse experiments with five and four replications, the control value of KYC 1126 on phyllosphere and rhizosphere was 88% and 36%, respectively. Whereas, the control value of dimetnomorph+propineb on phyllosphere was 100% and that of propamorcarb on rhizosphere was 44%. There was a phytotoxicity of the myxobacterial filtrate when seedlings were washed and soaked for 24 hours. Gummy materials were covered with roots. And stem and petiole were constricted, then a whole seedling was eventually blighted.

• Research in Plant Disease
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• v.22 no.4
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• pp.293-296
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• 2016

In late June 2016, stalk rot symptoms were observed on five vatieties of sorghum (Sorghum bicolar) at organic paddy-upland rotation system in Anseong city, Korea. The initial symptom on stalk surfaces was red color with a dark red spot lesion. A fungus was isolated from the initial lesion, and cultured on potato dextrose agar. Size of microconidia mostly extend to $5-19{\times}2-{\mu}m$ in culture, with 0-1 septa and macroconidia extend to $29-52{\times}3-4{\mu}m$ with 4-6 septa. Pathogenicity was investigated using conidial suspension spray to seedling of sorghum. After 3 days of inoculation, the dark red lesion was produced on stalks. On the basis of mycological characteristics, pathogenicity, and internal transcribed spacer (ITS) rDNA sequence analysis, this fungus was identified as Fusarium thapsinum. This is the first report of stalk rot on sorghum caused by F. thapsinum in Korea.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.139-147
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• 2002

Objective : This study was to establish a reproducible differentiation system from the parthenogenetic mouse embryonic stem (P-mES02) cells into functional cardiomyocytes like as in vitro fertilization mouse embryonic stem (mES01) cells. Materials and Methods: To induce differentiation, P-mES02 cells were dissociated and aggregated in suspension culture environment for embryoid body (EB) formation. For differentiation into cardiomyocytes, day 4 EBs were treated with 0.75% dimethyl sulfoxide (DMSO) for another 4 days (4-/4+) and then were plated onto gelatin-coated dish. Cultured cells were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. This frequency was compared with the results of DMSO not treated P-mES02 group (4-/4-) and mES01 groups (4-/4+ or 4-/4-). For confirm the generation of cardiomyocytes, beating cell masses were treated with trypsin-EDTA, dispersed cells were plated onto glass coverslips and incubated for 48 h. Attached cells were fixed using 4% paraformaldehyde and incubated with specific antibodies (Abs) to detect cardiomyocytes (anti-sarcomeric ? -actinin Ab, 1 : 100; anti-cardiac troponin I Ab, 1 : 2000) for 1 h. And the cells were finally treated with FITC or TRITC labelled 2nd Abs, respectively, then they were examined under fluorescence microscopy. Results: Rhythmically contracting areas in mES01 or P-mES02 cells were firstly appeared at 9 or 10 days after EBs plating, respectively. The highest cumulative frequency of beating EBs was not different in both treatment groups (mES01 and P-mES02, 4-/4+) with the results of 61.3 % at 13 days and 69.8% at 15 days, respectively. Also, the contracting duration of individual beating EBs was different from minimal 7 days to maximal 53 days. However, DMSO not treated groups (mES01 and P-mES02, 4-/4-) also had contracting characteristics although their frequency was a few compared to those of DMSO treated groups (6.0% and 4.0%). Cells recovered from the spontaneously contracting areas within EBs in both treated groups were stained positively with muscle specific anti-sarcomeric ? -actinin Ab and cardiac specific anti-cardiac troponin I Ab. Conclusion: This study demonstrated that the P-mES02 cell-derived cardiomyocytes displayed similarly structural properties to mES01 cell-derived cardiomyocytes and that the DMSO treatment enhanced the cardiomyocytes differentiation in vitro.

• Research in Plant Disease
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• v.26 no.1
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• pp.8-18
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• 2020

Red pepper, one of the major economic crops in Korea, is being affected by anthracnose disease caused by Colletotrichum acutatum. To control this disease, an antagonistic bacterial strain, Bacillus subtilis YGB36 identified by 16S rDNA sequencing, physiological and biochemical analyses is used as a biological control agent. In vitro screening revealed that the strain YGB36 possess strong antifungal activity against the pathogen Cylindrocarpon destructans. The strain exhibited cellulase, protease, amylase, siderophore production and phosphate solubility. In vitro conidial germination of C. acutatum was most drastically inhibited by YGB36 cell suspensions (10⁶ cfu/ml) or culture filtrate. Development of anthracnose symptoms was reduced on detached immature green pepper fruits by treatment with cell suspensions, and its control value was recorded as 65.7%. The YGB36 bacterial suspension treatment enhanced the germination rate of red pepper seeds and promoted root development and growth under greenhouse conditions. The in vitro screening of fungicide and insecticide sensitivity test against YGB36 revealed that the bacterial growth was not affected by any of the insecticides, and 11 fungicides out of 21 used. Collectively, our results clearly suggest that the strain YGB36 is considered as one of the potential biocontrol agents against anthracnose disease in red pepper.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.175-182
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• 1995

Rice is one of the most successful monocot in regenerating fertile and genetically stable transgenic plants. However there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper we first demonstrate that a Korean variety Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphotransferase) and GUS ($\beta$-glucuronidase) genes in separate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiment, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hy $g^{R}$ calli. The average efficiency of plantlet regeneration was apprbximately 27%. Based on the GUS activities of hygromycin resistant calli, ca.35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and Rl seeds were obtained. By examining the in siぉ activity of GUS in Rl seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the Rl generation.n.

• Journal of Food Hygiene and Safety
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• v.31 no.3
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• pp.216-221
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• 2016

This study was performed to evaluate the bactericidal efficacy of a fumigation disinfectant containing 35% paraformaldehyde against Salmonella Typhimurium (S. Typhimurium). In this study, the efficacy test of a fumigant against S. Typhimurium was carried out according to French standard NF T 72-281. The S. Typhimurium working culture suspension number (N value), all bacteria numbers on the carriers exposed to the fumigant (n1, n2, and n3), the number of bacterial suspensions by the pour plate method (N1), the number of bacterial suspensions by the filter membrane method (N2), and the mean number of bacteria recovered on the control carriers (T value), were obtained from the preliminary test. In addition, the reduction number of S. Typhimurium exposed to the fumigant (d value) was calculated using the T value, the mean number of bacteria in the recovery solution (n'1) and the mean number of bacteria on carriers plated in agar (n'2). The N value was $5.5{\times}10^8$ colony forming units (CFU)/mL, and n1, n2, and n3 were higher than 0.5N1, 0.5N2 and 0.5N1, respectively. Additionally, the T value was $3.5{\times}10^6CFU/carrier$. In terms of the bactericidal effect of the fumigant, the d value was 5.25. According to the French standard for fumigants, the d value for an effective bactericidal fumigant should be greater than 5. The results indicated that the fumigant containing 35% paraformaldehyde had an efficient bactericidal activity against S. Typhimurium, and, therefore, can be used to disinfect food materials and kitchen appliances contaminated with foodborne bacteria.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.101-106
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• 1990

These mxperiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro fertilization and following development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6mm of diameter. Bovine oocytes were matured in vitro for 24~26hrs in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$ and subsequently cultured in medium containing cumulus cell antibody for 1 hour. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hrs in BO solution 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and then cultured for 7 days. The results obtained in these experiments were summarized as follows : 1. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cells, the fertilization rate of cumulus intact and removed oocytes was ranged to 45.0 to 53.7%. These value is slightly lower than that(64.3%) of follicular oocytes not treated with the antibody, and increased frequency of both male and female pronuclear formation was found in cumulus intact oocytes cultred in medium without the antibody(p<0.05). 2. The fertilization rate of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cells was ranged 45.0 to 52.5%, significantly lowre than that(62.8%) of oocytes cultured in antibody free medium, and increased frequency of ova with male and female pronuclei was found when cumulus cells were present(p<0.05). 3. The rates of cumulus cell intact and removed oocytes developed to 8-, 16-cell and morula or blastocyst after treatment of intact and solubilized cumulus cell antibody were ranged 7.1 to 14.5, 2.9 to 5.9 and 1.5 to 2.9%, respectively, slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results of this stduy indicate that cumulus cells promote not only normal fertilization with proper pronuclear formation, but embryo development and that the beneficial effect of cumulus cell to the pronuclear formation and embryo development is blocked by the action of antibody to cumulus cell.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.183-187
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• 1999

To establish an efficient screening system for new herbicides using plant cultured cells, responses of tobacco photomixotrophic cultured (PH) cells to various herbicides with different modes of action were surveyed by measuring the cell growth and ion conductivity in medium. The cells were cultured in Murashige and Skoog (MS) medium containing 0.7mg/L 2,4-D, 0.3mg/L kinetin and 30 g/L sucrose at $25^{\circ}C$ in the light (100 rpm). Chemicals were treated to suspension cultures of tobacco PH cells at the time of subculture. The cell growth and ion conductivity in the medium were investigated on 12 days after chemical treatment. The ion conductivity assay gave well correlated results to the cell growth inhibition data. The responses of tobacco PM cells were dependent on the modes of action of chemicals tested. Atrazine, an inhibitor of photosynthetic electron transport (PET), strongly inhibited both the cell membrane and cell growth ($IC_{50}$/, about 1 $\mu$M). Butachlor (an inhibitor of cell division), glufosinate (an inhibitor of amino acid biosynthesis), and fluridone (an inhibitor of carotenoid biosynthesis) showed a dose-dependent inhibition. However, Quinclorac, a herbicide with an auxin activity, did not affect the cell growth and ion leakage. These results suggested that tobacco PM cells is suitable materials for the simple screening of new herbicides such as PET, amino acid biosynthesis, ceil division inhibitors by measuring the cell growth and ion conductivity.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.381-385
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• 2003

An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100％) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA＋0.5mg/L BA or 0.5 mg/L NAA＋0.5mg/L BA＋0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70％) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.