• Korean Journal of Plant Tissue Culture
• /
• v.27 no.3
• /
• pp.181-184
• /
• 2000

Proteins in friable and compact calli of Viola patrinii DC. were analysed. The protein contents in friable calli were lower than those in compact calli. In suspension culture, it increased to the maximum after 3 weeks culture from inoculation and decreased after 4 weeks culture. Several strong lavel signals were detected with the SDS-PAGE analysis. The polypeptides of 28, 31 and 35 KD were observed from the friable cell culture, from the compact cell culture strong band of 30 KD was determined, indicating that these polypeptides may be the major protein occurring during their cultures. Changes in amino acid contents during culture of the viola suspension cells were investigated the amino acid contents were greatest between two and three weeks culture of the viola suspension cells.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2022.09a
• /
• pp.62-62
• /
• 2022

Freesia has been an important worldwide cut flower because of its fragrance, long vase life and the wide color range of the flower. The conventional propagation methods by seeds and corms have many disadvantages such as shorter inflorescences with fewer numbers of florets, a reduction in cut flower quality and the accumulation of plant viruses in corms by successive cultivation. Therefore, the conventional propagation systems in Freesia needs to be replaced with tissue cultures to overcome the disadvantages. This study explored an efficient multiplication protocol using the combination of plant growth regulators (PGRs) for developed cultivar 'Sunny Gold'. The combination between 6-benzylaminopurin (BA) and α-naphthalene acetic acid (NAA) did not produce new shoots but developed enlarged roots. BA only treatments and the combination between BA and kinetin treatments were effective on shoot multiplication. The highest average number of shoots was 5.3 in the presence of 3 mg/L BA and 0.5 mg/L kinetin. To produce corms and cormlets, proliferated shoots were subcultured on 1/2 Murashige and Skoog (MS) medium supplemented with 90 g/L sucrose, 1 g/L charcoal and 7 g/L plant agar and placed at 4℃ in the dark for 6 months. The small size of corms and comlets were produced. The average number of regenerated comlets was 2.75 per shoot. The results showed that shoot multiplication is more efficient than cormlet regeneration for in vitro freesia proliferation.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.1
• /
• pp.23-28
• /
• 1994

This study was rimed out to examine the difference in the cell vitality between mesophyII protoplast (MP) and paraveinal mesophyII protoplast (PVMP) of Nicotiana tabaccum 'Xanti', Petunia hybrida 'Blue Star' and Chrysanthemum morifolium 'Baeckwang' by using urea permeability technique. The effects of various enzyme solutions and incubation time, NAA and thidiazron on plant regeneration from isolated protoplasts were also investigated. The vibratome technique was used for protoplast isolation and urea permeability test because the fresh living, thin tissue stripes (50 ${\mu}{\textrm}{m}$ of thickness) could be obtained with minimal damage with the vibratome. For the three plants examined, the urea permeability on the tested tissue stripes was relatively higher in PVMP than in MP by about Ks = 2.0 $\times$ 10$^{-5}$ cm/sec. The treatment of an enzyme mixture of 1.5% cellulase R-10, 1% Driselase, 0.5% Macerozyme R-10, and 0.5% Pectinase for 4 to 8 h was effective on the isolation of PVMP. The highest frequency of callus formation and plant regeneration from the isolated protoplasts was obtained with NAA 2 mg/L and thidiazuron 0.01 mg/L. Furthermore, the results demonstrated that cell devision and plantlet regeneration was more frequent in the PVMP than in the MP of the same leaf or plant We, therefore, conclude that UM is an excellent experimental material for the callus formation and regeneration from isolated protoplasts.

• Journal of Korean Society of Forest Science
• /
• v.94 no.1 s.158
• /
• pp.39-44
• /
• 2005

An effective micropropagation system for Liriodendron tulipifera L via somatic embryogenesis was established using immature seeds. Immature seeds from five individual trees were bisected longitudinally and cultured on two basal media (MS and B5) containing different combinations of 2,4-D and TDZ to induce callus and embryogenic tissue under light ($40{\mu}mol\;m^{-2}s^{-1}$, 16 hr/day) or complete darkness at $25{\pm}2^{\circ}C$. There was no distinctive difference on callus and embryogenic tissue induction between the two basal media with PGRs. Optimum culture medium appeared to be MS medium supplemented with 1.0mg/L 2,4-D and 0.01mg/L TDZ plus 3% sucrose. Nonembryogenic callus induction rate was not significantly different among the genotypes. However, However, the embryogenic callus induction frequency differed greatly by the genotypes ranging from 55% to 72% when cultured in the dark. Generally, the cultures maintained in the dark tended to show normal somatic embryo development as well as embryogenic tissue formation and this was confirmed by histological examination. Above results suggest that a proper selection of mother tree and dark culture condition are necessary to optimize somatic embryogenesis system of Liriodendron tulipifera.

• Korean Journal of Veterinary Research
• /
• v.33 no.2
• /
• pp.269-275
• /
• 1993

Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained with primary antiserum and conjugated second antibodies as recommended by manufacturer. None of the cultures tested showed virus-induced phenomena. Immunoreactive products were commonly found in the liver, in some cases there were also positive staining in the spleen and kidneys. Other organs showed weak or insignificant immunoreactions. By ABC method on the formalin-fixed, paraffin-embedded liver tissues, strong immunoreactivity was found in the periportal triad lesions and peripheral lesions of the hepatic lobules. Immunoreactive products showed diffuse fine granular in the cytoplasm of hepatocytes and sinusoidal cells. In some cells, immunoproducts marginate at the periphery of the cells. The intensive staining of the cytoplasm of infected cells allowed their exact differentiation from surrounding uninfected cells. The positive area involved coincided with histopathological lesion on serial liver sections. In conclusion, liver was proved to be a consistent target organ in RHD, and the immunoperoxidase method in the section of formalin-fixed, paraffin-embedded hepatic tissue could be broadly used for the routine diagnosis of the disease.

• Korean Journal of Clinical Laboratory Science
• /
• v.36 no.2
• /
• pp.163-172
• /
• 2004

Mast cells (MCs) have been implicated in the pathogenesis of tissue fibrosis. However, the role of MC in the development of liver fibrosis has not been fully elucidated. Stem cell factor (SCF) is known to recruit MCs to the liver following injury as it induces mast cell proliferation, survival and differentiation from resident tissue precursors. This study examines the interaction between activated hepatic stellate cells (HSCs) and MCs in rat fibrotic liver, and SCF production by HSCs during culture in vitro. Rats were studied 4, 7, 14 and 21 days after bile duct ligation (BDL). Fibrogenesis was assessed by a measurement of collagen stained with sirius red F3B. Activated HSCs and MCs were identified by ${\alpha}$-smooth muscle actin (${\alpha}-SMA$) immunohistochemical and alcian blue staining and measured by a computerized image analysis system. SCF production was determined in rat HSC cultures using Western blotting. Mild fibrotic changes were noted in BDL rat livers as early as 4 days after induction of cholestasis. Significant expansion and organization of fibrous tissue has occurred in day 14 BDL rats which progressed to bridging fibrosis by day 21. In BDL rats, both a large number of activated HSCs and MCs were detected in portal tracts and fibrous septa. Both area of activated HSCs infiltration and density of MCs were significantly higher in all BDL group compared with Shams. In BDL rats, both areas of activated HSCs infiltration and density of MCs were no significant difference between day 4 and 7 and were significantly higher in day 14. However, the areas of activated HSCs infiltration were significantly lesser in day 21 and the densities of MCs were significantly higher in day 21 compared with day14 BDL. In BDL rats, both areas of activated HSCs infiltration and density of MCs were highly correlated with areas of fibrosis. Western blotting showed that SCF protein was consistently produced in activated HSCs by culture on plastic and freshly isolated HSCs expressed relatively little 30kD SCF compared to late primary culture activated HSCs (day 14) and passaged HSCs. These results suggest that HSCs activated in vitro produce SCF, and may play an important role in recruiting mast cells to the liver during injury and fibrosis.

• The Journal of Korean Academy of Prosthodontics
• /
• v.41 no.3
• /
• pp.300-318
• /
• 2003

Statement of problem: The success of implants depends on intimate and direct contact of implant material on bone tissue and on functional relationship with soft tissue contact. Creation and maintenance of osseointegration depend on the understanding of the tissue's healing, repairing, and remodeling capacity and these capacities rely on cellular behavior. Altering the surface properties can modify cellular responses such as cell adhesion, cell motility, bone deposition, Therefore, various implant surface treatment methods are being developed for the improved bone cell responses. Purpose: The purpose of this study was to evaluate the responses of osteoblast-like cells to surface-modified titanium. Materials and Methods: The experiment was composed of four groups. Group 1 represented the electropolished surface. Group 2 surfaces were machined surface. Group 3 and Group 4 were anodized surfaces. Group 3 had low roughness and Group 4 had high roughness. Physicochemical properties and microstructures of the discs were examined and the responses of osteoblast-like cells to the discs were investigated. The microtopography was observed by SEM. The roughness was measured by three-dimension roughness measuring system. The microstructure was analyzed by XRD, AES. To evaluate cell responses to modified titanium surfaces, osteoblasts isolated from calvaria of neonatal rat were cultured. Cell count, morphology, total protein measurement and alkaline phosphatase activities of the cultures were examined. Results and Conclusion: The results were as follows 1. The four groups showed specific microtopography respectively. Anodized group showed grain structure with micropores. 2. Surface roughness values were, from the lowest to the highest, electropolished group, machined group, low roughness anodized group, and high roughness anodized group. 3. Highly roughened anodized group was found to have increased surface oxide thickness and surface crystallinity. 4. The morphology of cells, flattened or spherical, were different from each other. In the electropolished group and machined group, the cells were almost flattened. In two anodized groups, some cells were spherical and other cells were flattened. And the 14 day culture cells of all of the groups were nearly flattened due to confluency. 5. The number of attached cells was highest in low roughness anodized group. And the machined group had significantly lower cell count than any other groups(P<.05). 6. Total protein contents showed no difference among groups. 7. The level of alkaline phosphatase activities was higher in the anodized groups than electropolished and machined groups(P<.05).

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.1
• /
• pp.57-61
• /
• 2000

We investigated the levels of glutathione (GSH) and its oxidized form (GSSG) in 24 cell lines derived from various plant species to understand the antioxidative mechanism in plant cell cultures. The total glutathione content was 98$\pm$27 $\mu$g/g cell fresh wt, showing a slight difference in plant species. The average contort of GSH and GSSG was 72$\pm$20 and 26$\pm$10 $\mu$g/g cell fresh wt, respectively. The average GSH content in plant cell lines occupies approximately 73% in total glutathione. During the suspension cultures of Scutellaria baicalensis, one of the plant species we tested, the GSH content decreased in proportion to the cell growth during the exponential growth stage, showing the low level at the stationary growth stage (84 $\mu$g/g cell fresh wt), whereas the GSSG content increased to the stationary growth stage (31 $\mu$g/g cell fresh wt). The results suggested that the ratio of GSH and GSSG should be involved in the cell growth and antioxidative mechanism in cultured cells.

• Korean Journal of Plant Tissue Culture
• /
• v.26 no.4
• /
• pp.259-264
• /
• 1999

In order to study the role of ethylene on the formation of adventitious roots, trichomes and calli, the effects of 1-aminocyclopropane-l-carboxylic acid (ACC), ethephon, $CoCl_2$ and $AgNO_3$ were investigated in the leaf segments from ecotype Columbia of Arabidopsis thaliana. When the leaf segments were cultured on the media for forming adventitious roots (0.1 mg/L NAA), trichomes (2.0 mg/L NAA) and calli (10.0 mg/L NAA), and then each cultures was treated with 1-100 mg/L of ACC and ethephon, respectively. On the adventitious root-forming medium adventitious root formation was decreased, and trichomes were induced. And on the trichome-forming medium trichome formation was decreased, and calli were induced. In order hand each culture was treated with 1-100 mg/L of $CoCl_2$ and $AgNO_3$, respectively. On the adventitious root-forming medium adventitious roots was increased without trichome formation, and on the trichome-forming medium trichome formation was decreased, and adventitious roots were induced. However on the callus-forming medium treated with ACC, ethephon, $CoCl_2$ and $AgNO_3$, respectively, callus formation was inhibited and trichomes were induced in all cultures.

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.3
• /
• pp.203-211
• /
• 2000

In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.