• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Korean Journal of Microbiology
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• v.53 no.4
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• pp.347-350
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• 2017

We sequenced the genome of the Sphingobacteriaceae bacterium SH-48 isolated from the Sohan stream in Republic of Korea by using a dilution-to-extinction culturing method. The sequences were assembled into a draft genome containing 5,650,162 bp with a G + C content of 35.4% and 4,856 protein-coding genes in 2 contigs. This strain contains the carotenoid biosynthesis genes crtY, crtZ, crtD, crtI, crtB, and crtH as gene clusters. This genomic information provides new insights into the carotenoid biosynthesis pathway.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.212-226
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• 2002

North American ginseng production may have been maximized in the traditional growing areas in the last decade and further increases may be in woods grown root, for niche markets. The marketplace demands high quality roots. Most problems leading to low quality roots start with the grower and can be avoided. These include poor site selection, inadequate soil drainage, untimely and poorly applied pesticides, and neglect of good sanitary practices. Selection of low lying sites increased the plant damage from frost in Ontario in May 2002. Seeding is still the major method of propagation of ginseng in spite of some success in culturing different parts of the plant. Opportunities exist for shortening the stratification period of North American ginseng seed to allow spring planting. This may reduce disease incidence. Since only one-third of ginseng seed sown ultimately produces plants harvested after 3 years any approach that reduces disease incidence and improves seed germination, seedling emergence and crop stand must be pursued. Disease is the major problem in ginseng cutivation from seed stratification, soil preparation prior to planting, right through to drying of the roots. Replant disease remains as an unresolved problem and needs full characterization and new approaches for control. Much progress has been made in research and related extension activities in disease control although challenges will arise such as with Quintozene and its replacement with Quadris for control of diseases caused by Rhizoctonia. Decreased labor populations and increased associated costs for ginseng production are causing rapid mechanization in every aspect of the ginseng industry. Engineers, machinery dealers, and fabricators, and growers are being challenged to increase efficiency by mechanization.

• Journal of Life Science
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• v.8 no.1
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• pp.8-13
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• 1998

The maize mitochondrial mutant (NCS2) is derived from homologous recombination between genes encoding NADH dehydrogenase subunit 4 and subunit 6. Plants from mitochondria mutants exhibited severe related growth and development including dwarfism and striping on the leaves. Aborted embryos from NCS2 mutants have been rescued and cultured on the N6 medium supplemented with 2,4-D 1 mg/l. Most calli from NCS2 aborted embryos showed slow growing pattern at first stage. However, upon continuous culturing them on the medium, those were segregated into mutant and normal callus lines. These segregations could be detected by using PCR method with three primers. Such segregation seems to be resulted from the preferential growth of normal cells over the mutant cells on the normal culture condition. Therefore, this method can be used for determining rate of indirect cytoplasmic segregation by estimating amplified band intensities. When NCS2 mutant callus lines cultured on regeneration medium, no adventitious shoot induction was observed. However, callus lines with more mitochondria induced adventitious shoots. These studies suggest that mitochondria NADH-dehydrogenase for electron transport in the inner membrane of mitochondria is essential for the differentiation and development of plants.

• Korean Journal of Plant Resources
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• v.31 no.6
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• pp.684-694
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• 2018

This study describes the successful establishment of a cryopreservation protocol for Citrus limon cultivars: 'Frost Eureka limon' and 'Cook Eureka limon', using a droplet-vitrification method. The shoot tips that were excised from in vitro grown seedlings of the two cultivars were preserved in liquid nitrogen (LN) and successfully regenerated into whole plants. Excised shoot tips were pre-cultured for 1 or 2 days in 0.3 M and 0.5 M sucrose solutions at $25^{\circ}C$ and incubated in a loading solution (LS) composed of 17.5% glycerol + 17.5% sucrose in Murashige and Skoog (MS) medium for 40 min at $25^{\circ}C$. Prior to direct immersion in LN for 1 h, the shoot tips were dehydrated with plant vitrification solution 2 (PVS2) at $0^{\circ}C$ or PVS3 at $25^{\circ}C$. The frozen shoot tips were re-warmed and unloaded with 1.2 M sucrose in $\text\tiny{^1/_2}$ MS for 30 min at $25^{\circ}C$. Shoot tips were post-cultured overnight on survival medium and then micrografted onto 'trifoliate orange' (Poncirus trifoliate (L.) Raf. seedling rootstocks for recovery and to produce whole plants. The highest regrowth rates were 53.5% and 50.3% for cryopreserved shoot tips of 'Frost Eureka limon' and 'Cook Eureka limon', respectively, when pre-cultured in 0.3 M and 0.5 M sucrose concentrations in a sequencing manner, with LS and treated with PVS2 for 60 min at $0^{\circ}C$. We also investigated whether the ammonium ion concentration on post-culture medium affected the viability of the cryopreserved Citrus shoot tips. The viability of cooled samples, following culturing on woody plant media (WPM) containing $\text\tiny{^1/_4}$ ammonium nitrate overnight before micrografting, was the highest (70.3%) in 'Frost Eureka limon'. The study described here is a cost-effective and safe method to conserve Citrus fruit cultivars, for the improvement and large-scale multiplication of fruit plants and for breeding disease resistance.

• Korean Journal of Medicinal Crop Science
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• v.28 no.6
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• pp.445-454
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• 2020

Background: Culturing wild ginseng adventitious root using plant factory technology provides genetic safety and high productivity. This production technology is drawing attention in the fields of functional raw materials and product development. The cultivation method using elicitors is key technology for controlling biomass and increasing secondary metabolites. Methods and Results: Elicitor treatments using methyl jasmonate, pyruvic acid, squalene, β-sistosterol were performed to amplify total ginsenosides (Rb1, Rc, Rb2, Rb3, and Rd) of cultured wild ginseng adventitious root. Thereafter, fermentation and steaming processes were performed to convert total ginsenosides into minor molecular ginsenosides (Rg3, Rk1, and Rg5). The result indicated that methyl jasmonate minimizes the reduction in fresh weight of cultured wild ginseng adventitious root and maximizes total ginsenosides (sum of Rb1, Rc, Rb2, Rb3, and Rd). Ginsenoside conversion results showed a maximum degree of conversion of 131 mg/g. Conclusions: In this study, we demonstrated that the optimal elicitor treatment method increased the content of total ginsenosides, while the steaming and fermentation processing method increased the content of minor ginsenosides.

• Journal of Korean society of Dental Hygiene
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• v.17 no.2
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• pp.319-330
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• 2017

Objectives: The purpose of this study was to compare SYBR Green qPCR, TaqMan, and bacterial selective medium cultures for accurate quantitative analysis of oral microorganisms. Methods: The SYBR Green method is widely used to analyze the total amount of oral microorganisms in oral saliva. However, in this study, MTR-PCR method based on TaqMan method was performed using newly developed primers and probes. In addition, it was designed to confirm the detection agreement of bacteria among bacteria detection method. Results: As a result of MRT-PCR and SYBR Green qPCR analysis, more than 40 times (0.9-362.9 times) bacterium was detected by MRT-PCR. In addition, more bacteria were detected in saliva in the order of MRT-PCR, SYBR Green qPCR, and bacterium culture, and the results of MRB-PCR and SYBR Green qPCR showed the highest agreement. The agreement between the three methods for detecting P. intermedia was similar between 71.4 and 88.6%, but the agreement between MRT-PCR and SYBR Green qPCR was 80% for S. mutans. Among them, the number of total bacteria, P. intermedia and S. mutans bacteria in saliva was higher than that of SYBR Green qPCR method, and bacterium culture method by MRT-PCR method. P. intermedia and S. mutans in saliva were detected by MRT-PCR and MRT-PCR in 88.6% of cases, followed by the SYBR Green qPCR method (80.0%). Conclusions: The SYBR Green qPCR method is the same molecular biology method, but it can not analyze the germs at the same time. Bacterial culturing takes a lot of time if there is no selective culture medium. Therefore, the MRT-PCR method using newly developed primers and probes is considered to be the best method.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.417-422
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• 2008

Apicidin is a cyclic tetrapeptide produced by some Fusarium species and is known to inhibit Apicomplexan histone deacetylase. The goals of this study were to determine species identity of Fusarium isolate KCTC16676, an apicidin producer, to improve a method for apicidin extraction, and to test phytotoxicity of apicidin and its analogs. We compared sequences of the translation elongation factor 1-alpha (TEF) gene in KCTC16676 with those from isolates representing diverse Fusarium species, which showed that KCTC16676 belongs to the F. semitectum-F. equiseti species complex. To enhance apicidin production, after culturing isolate KCTC16676 on a wheat medium for 3 weeks at $25^{\circ}C$, the culture was extracted with chloroform. Apicidins were purified through a reverse phase $C_{18}$ silica gel column, resulting in 5 g of apicidin, 200 mg of apicidin A, and 300 mg of apicidin $D_2$ from 4 kg of wheat cultures; this represents a significant yield improvement from a previous method, offers more materials to study the modes of its action, and facilitates the elucidation of the apicidin biosynthesis pathway. Apicidin and apicidin $D_2$ showed phytotoxicity on both seedlings and 2-week-old plants of diverse species, and weeds were more sensitive to apicidins than vegetables

• Journal of Animal Science and Technology
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• v.44 no.4
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• pp.491-498
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• 2002

The inhibitory effect of lactobacilli and bifidobacteria on the growth of typical intestinal pathogens, Escherichia coli and Salmonella typhimurium was studied. The degree of inhibition was measured by well disc assay and turbidimetry method. The strains which showed the higher antimicrobial activity were L. acidophilus La-5, L. acidophilus NCFM, L. casei Lc-01 on the average by using two different methods. The associative cultures were performed with selected 3 lactobacilli and 2 enteropathogens E. coli and S. typhimurium, respectively. Inhibition of pathogen began at 9hr after culturing so that viable counts was decreased rapidly. After 30hr incubation, there were no viable pathogens from the mixed culture. Under this experimental condition, the antimicrobial activity of lactic acid bacteria was not due to pH alone and supposed to different to the strains.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.53.1-53.1
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• 2011

Bioactive glass powders were synthesized by hydrothermal chemical route by the use of ultrasonic energy irradiation. We used sodalime, calcium nitrate tetra hydrate and di ammonium hydrogen phosphate as the precursor material to synthesize $SiO_2$ rich bio-active glass materials. The $SiO_2$ content was varied in the precursor mixture to 60, 52 and 45 mole%. Dense compacts were obtained by microwave sintering at $1,100^{\circ}C$. Mechanical properties were characterized for the fabricated dense bioactive glasses and were found to be comparable with conventional CaO-$SiO_2$-$Na_2O$-$P_2O_5$ bioactive glass. Detailed biocompatibility evaluation of the glass composition was investigated by in-vitro culture of MG-63 cell and mesenchyme stem cell. Cell adhesion behavior was investigated for both of the cell by one cell morphology for 30, 60 and 90 minutes. Cell proliferation behavior was investigated by culturing both of the cells for 1, 3 and 7 days and was found to be excellent. Both SEM and confocal laser scanning microscopy were used for the investigation. Western blot analysis was performed to evaluate the bimolecular level interaction and extent and rate of specific protein expression. The ability to form biological apatite in physiological condition was observed with simulated body fluid (SBF). In-vivo bone formation behavior was investigated after implanting the materials inside rabbit femur for 1 and 3 month. The bone formation behavior was excellent in all the bioglass compositions, specially the composition with 60% $SiO_2$ content showed most promising trend.