• Applied Biological Chemistry
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• v.38 no.5
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• pp.435-439
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• 1995

Some characteristics and pathogenicity of Hyphantria cunea nuclear polyhedrosis virus (HcNPV), a potential microbial pesticide was studied. H. cunea NPV replicated in the nucleus of S. frugiperda cells cultured in the TNMFH medium. In case of virus infected cell, prepolyhedra formation was observed at 24hrs post-infection. At 48 hrs post-infection, Most of the infected cell contained many mature polyhedra which were released into culture media 72 hrs post-infection, with the cells grown in suspension culture, pH of the culture medium increased during the virus replication: the pH of fresh medium was 6.35 and rose to 6.77 within 120 hrs. Polyhedra formed a band in linear density gradient of sucrose by centrifugation, which co-sedimented with $50{\sim}55%$ sucrose. The shape of the purified polyhedra was mostly tetragonal hexahedron and its size was about $2.5{\mu}m$. Electron microscopy and phase contrast microscopy showed that many bundled nucleocapsids were occluded in mature polyhedra at 48 hrs post infection. H. cunea larvae infected with NPV showed a higher motality in the second and third instar than in the fourth instar. Death rate of H. cunea larvae in the second and third instar fed with leaves coated with $1.5{\times}10^{9}{\sim}l.5{\times}10^{7}PIBs/ml$ reached more than 90%.

• Korean Journal of Agricultural Science
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• v.22 no.1
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• pp.54-61
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• 1995

In order to investigate the effects of Ganoderma lucidum on the fermentation of media by Kluyveromyces fragilis KCCM 35458, with 0%, 0.1%, 0.5%, and 1.0% (w/v) of Ganoderma lucidum extract were added to skin milk and fermented in the suspension culture at $30^{\circ}C$ for 72 hours. The pH, acidity, $CO_2$ evolution, alcohol production, number of yeast cells and lactose content were investigated. The results are summarized as follows; 1. As fermentation time advanced by the pH was decreased and the acidity was increased in the media of 1.0% Gnaoderma lucidum extract as fermentation time advanced. 2. $CO_2$ evolution was increased in the all of Genoderma lucidum extract and significantly effect showed in 1.0% extract medium. 3. During 72 hours culturing, alcohol was detected in all Gandoderma lucidum extract media, 1.0% Ganoderma lucidum extract medium showed extremly high in alcohol and shortened fermentation time. 4. The number of yeast was increased more at logarithmic phase and the growth of yeast was more rapid in all Ganoderma lucidum extract especially 0.5%, 1.0% media samples than control. 5. As fermentation time goes by, the lactose content of media were decreased more rapidly in 1.0% Ganoderma lucidum extract medium than control.

• FLOWER RESEARCH JOURNAL
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• v.17 no.2
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• pp.93-100
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• 2009

This study was carried out to investigate the efficient in vitro mass propagation methods for juvenile sporophytes of Matteuccia struthiopteris. Chopped segments of pinnae, petiole and rhizome were cultured on 1/2MS with 0.1% activated charcoal. Among these explant sources only rhizome segments produced young sporophytes, regenerating vigorously on 1/2 MS medium. Adjusting sucrose concentration to 2% and supplement to $50mgL^{-1}$ $NaH_2PO_4$ in 1/2MS medium proved to be more efficient for plant regeneration. Various combinations of growth regulators such as kinetin, BA, NAA, and IBA were added to the growing media, and the best sporophyte regeneration was obtained by $1{\mu}M$ kinetin. The BA addition resulted in vigorous proliferation of meristematic tissues, but without differentiation to sporophytes. Three types of culture methods, solid using agar, liquid stationary, and liquid shaking culture, were employed with or without activated charcoal. The addition of 0.1% activated charcoal to modified 1/2MS media (2% sucrose, $50mgL^{-1}$ $NaH_2PO_4$, $1{\mu}M$ kinetin, pH 5.8 and 0.8% agar) yielded highest sporophyte regeneration in liquid shaking culture.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.7-13
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• 2001

Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.163-169
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• 2001

OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P ＜ 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

• Development and Reproduction
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• v.12 no.1
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• pp.87-95
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• 2008

Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.

• Research in Plant Disease
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• v.22 no.1
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• pp.18-24
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• 2016

This study was conducted to elucidate the cultural and pathogenic characteristics of Cladosporium cucumerinum PT1 and resistance of 81 commercial cucumbers (Cucumis sativus). Cucumber leaves and fruits appeared as scab were collected from a plastic film house located in Pyeongtaek, Gyeonggi-province, Korea in late March, 2015. A casual fungus was isolated from the diseased fruits on potato dextrose agar and it was identified as C. cucumerinum PT1 based on the morphological characteristics. To find out the effect of wounding and fruit size on the development of cucumber scab, small (<10 cm long), medium (10 to 20 cm long), and large (>20 cm long, commercially mature fruit) size cucumber fruits were harvested, C. cucumerinum PT1 pathogens were inoculated with a single droplet of suspension ($1{\times}10^5$ spores/ml) on wounded or unwounded cucumber fruits. Small fruits were completely damaged with showing severe water-soaking symptoms and fast pathogen growth regardless of wounded or unwounded. Meanwhile slight water-soaking symptoms on medium and large size fruits occurred and disease development into plant tissues was observed only on wounded fruits. Disease resistance of 81 commercial cucumber cultivars was evaluated on third-stage seedlings and small fruits by inoculating suspension ($1{\times}10^5$ spores/ml) of C. cucumerinum PT1. As a result, mini and pickling cultivar groups were resistant, 'Cheoeumcheoreom' cultivar was symptomless and the other cultivars were resistant to medium resistant. On the other hand, most of cucumber cultivars belonging to the other groups were susceptible. Disease resistance of cucumber against cucumber scab was significantly different among cultivars and a few cucumber cultivars showed different disease resistant responses to two bioassay methods using seedlings and small fruits. Therefore, to screen scab resistance in cucumber, a test using both fruits and seedlings is advisable. We think that the selected resistant cultivars can be used to control cucumber scab effectively under the farmhouse condition.

• Journal of Plant Biology
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• v.28 no.1
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• pp.69-77
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• 1985

The degree of The degree of The degree of ${Zn}^{2+}$ effect on the photosynthetic electron transport and photophosphorylation activities in barley chloroplasts has been tested.${Zn}^{2+}$treatment was done in the 2 ways. One was that it was added into the chloroplasts suspensions isolated from the plants grown under the normal ${Zn}^{2+}$level (10$^{-6}$ M). The other was that the different concentrations of ${Zn}^{2+}$was applied in each growth medium. Then, it was not added into the chloroplasts suspensions isolated from the plants. PS II activity in both way of the treatments was more severely inhibited than PS I by the increment of ${Zn}^{2+}$ concentration. The photophosphorylation activity measured by pH measurement was gradually decreased with the increase of ${Zn}^{2+}$concentration in both ways, too. However, it was shown that M $n^{2+}$ could be near fully overcome the inhibitory effect of ${Zn}^{2+}$in PS II, and $Mg^{2+}$ could also reduce the Z $n^{2+}$ inhibition in the photophosphorylation. In the low concentrations of $Mg^{2+}$ (3 to 5$\times$10$^{-3}$ M) in the suspension, ${Zn}^{2+}$(2$\times$10$^{-5}$ M) could increase the activity of photophosphorylation. As compares to other cations, Z $n^{2+}$ caused less inhibitory effect on the photophosphorylation activity than Cu, Cd, but more than Pb and Ni. It may be assumed that a complex from reaction of Z $n^{2+}$ and mercaptoethanol was produced and it could reduce the stability of CPI band during SDS-PAGE.effect on the photosynthetic electron transport and photophosphorylation activities in barley chloroplasts has been tested. Z $n^{2+}$ treatment was done in the 2 ways. One was that it was added into the chloroplasts suspensions isolated from the plants grown under the normal Z $n^{2+}$ level (10$^{-6}$ M). The other was that the different concentrations of Z $n^{2+}$ was applied in each growth medium. Then, it was not added into the chloroplasts suspensions isolated from the plants. PS II activity in both way of the treatments was more severely inhibited than PS I by the increment of Z $n^{2+}$ concentration. The photophosphorylation activity measured by pH measurement was gradually decreased with the increase of Z $n^{2+}$ concentration in both ways, too. However, it was shown that M $n^{2+}$ could be near fully overcome the inhibitory effect of Z $n^{2+}$ in PS II, and $Mg^{2+}$ could also reduce the Z $n^{2+}$ inhibition in the photophosphorylation. In the low concentrations of $Mg^{2+}$ (3 to 5$\times$10$^{-3}$ M) in the suspension, Z $n^{2+}$ (2$\times$10$^{-5}$ M) could increase the activity of photophosphorylation. As compares to other cations, Z $n^{2+}$ caused less inhibitory effect on the photophosphorylation activity than Cu, Cd, but more than Pb and Ni. It may be assumed that a complex from reaction of Z $n^{2+}$ and mercaptoethanol was produced and it could reduce the stability of CPI band during SDS-PAGE.effect on the photosynthetic electron transport and photophosphorylation activities in barley chloroplasts has been tested. Z $n^{2+}$ treatment was done in the 2 ways. One was that it was added into the chloroplasts suspensions isolated from the plants grown under the normal Z $n^{2+}$ level (10$^{-6}$ M). The other was that the different concentrations of Z $n^{2+}$ was applied in each growth medium. Then, it was not added into the chloroplasts suspensions isolated from the plants. PS II activity in both way of the treatments was more severely inhibited than PS I by the increment of Z $n^{2+}$ concentration. The photophosphorylation activity measured by pH measurement was gradually decreased with the increase of Z $n^{2+}$ concentration in both ways, too. However, it was shown that M $n^{2+}$ could be near fully overcome the inhibitory effect of Z $n^{2+}$ in PS II, and $Mg^{2+}$ could also reduce the Z $n^{2+}$ inhibition in the photophosphorylation. In the low concentrations of $Mg^{2+}$ (3 to 5$\times$10$^{-3}$ M) in the suspension, Z $n^{2+}$ (2$\times$10$^{-5}$ M) could increase the activity of photophosphorylation. As compares to other cations, Z $n^{2+}$ caused less inhibitory effect on the photophosphorylation activity than Cu, Cd, but more than Pb and Ni. It may be assumed that a complex from reaction of Z $n^{2+}$ and mercaptoethanol was produced and it could reduce the stability of CPI band during SDS-PAGE.ld reduce the stability of CPI band during SDS-PAGE.

• The Korean Journal of Rheology
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• v.10 no.4
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• pp.195-201
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• 1998

We have studied the rheological and electrical properties of two types of electrorheological (ER) fluids based on semi-conductive polymers (poly(p-phenylene) and polyaniline). These semi-conductive polymer-based suspensions showed a dramatic increase in viscosity on the application of the static electric field due to the large value of conductivity ratio between particle and medium. The dynamic yield stresses of these ER suspensions exhibited a quadratic dependence on electric field strength at low electric fields and a linear one for high fields. They showed a maximum and then decreased with increasing bulk conductivity of particles. These yield stress behaviors under the static electric field were found to be closely related to the dielectric properties, which is in accord with Maxwell-Wagner interfacial polarization induced by the conductivity effects. In order to achieve better understanding of interfacial polarization effect on ER response and to improve the stability of ER suspension, different kinds of surfactants were employed for controlling the ER activity as well as for enhancing the colloidal stability of suspensions.

• Korean Journal of Animal Reproduction
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• v.18 no.4
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• pp.235-244
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• 1995

The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.