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Free Radical Scavenging Activity and Ascorbate Content in Various Plant Cell Lines (다양한 식물배양세포주에서 자유라디칼 포착활성과 Ascorbate 함량)

  • AHN, Young-Ock;CHOI, Yong-Hwa;KWON, Seok-Yoon;LEE, Haeng-Soon;KIM, Suk-Won;PARK, Il-Hyun;KWAK, Sang-Soo
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.289-293
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    • 1998
  • We investigated the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in methanol extracts of 64 cultured cell lines, which were derived from various plant species, and the ascorbate content in cell lines, which showed a high radical scavenging activity. Thirteen cell lines revealed the antioxidative activity ($IC_{50}$) by methanol extracts of less than 50 mg in cell fresh wt. Of them, six cell lines showed the same Rf value as ascorbate on the DPPH sprayed silica gel TLC. The ascorbate content in cell lines of Rosa multiflora, Scutellaria baicalensis, and Achyranthes japonica showed 48.5, 30.3, and $16.8\;\mu\textrm{g}$ per g cell fresh wt by HPLC analysis, respectively. In callus cultures of S. baicalensis, the concentration of ascorbate reached a maximun ($39{\pm}3.4\;\mu\textrm{g}/g$ cell fresh wt) on 30 days after subculture, which corresponded to the stationary growth phase, and subsequently decreased by successive culturing.

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Callus Formation and Plant Regeneration from Immature Embryos of Eleutherococcus senticosus (가시오갈피 미숙배 배양으로부터 Callus 형성 및 식물체 재분화)

  • Yu, Chang-Yeon;Kim, Jae-Kwang;Ahn, Sang-Deuk
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.49-55
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embryos in Eleutherococcus senticosus. Immature embryos from seeds were removed under the microscope and placed on MS, $MSB_5\;and\;B_5$ medium containing several plant growth regulators. While the calli were well formed on media containing 2 mg/l of 2, 4-D, 2 mg/l of 2, 4-D and 0.7 mg/l of TDZ, shoot regeneration was better on MS medium with combinations of high concentrations of TDZ and low concentrations of 2, 4-D. Treatment of 2, 4-D alone was better than treatment of TDZ alone in callus induction, but plant regeneration was reversed. The results of callus formation and shoot regeneration on $MSB_5\;and\;B_5$ media were similar to those of MS media. The rate of callus formation was nearly 100% when 2, 4-D was added to $B_5$, medium on concentration of 2 mg/l or 0.7 mg/l. TDZ showed very significant effect on the formation of multiple shoots.

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Propagation of Cutting Method of a Rare Endemic Juniperus chinensis var. sargentii Henry in Korea (희귀수종 눈향나무(Juniperus chinensis var. sargentii Henry)의 삽목증식)

  • Song, Jeong-Ho;Jang, Kyung-Hwan;Hur, Seong-Doo
    • Korean Journal of Plant Resources
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    • v.23 no.4
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    • pp.368-373
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    • 2010
  • Juniperus chinensis var. sargentii Henry is a short and creeping evergreen shrub which reaches about 60 cm in height and only occurs in the northeast Asia and in the top of high mountains over the Korea. The Korea Forest Service protects it strictly by law since J. chinensis var. sargentii is an eccentric plant and possibly may be exterminated soon in Korea. This study was carried out to develop the propagation technique by cutting for conservation of genetic resources of J. chinensis var. sargentii. The rooting responses of branch cuttings, obtained from hard(May) and semi-hard wood shoots (August) to four growth regulators, namely, IAA, IBA, NAA and Rooton(exceptionally powder method) applied at various concentrations(0, 100, 200, 500, 1000 and 2000 $mgL^{-1}$) were examined in mixed soil media. Rooting rate showed significant difference between cutting times, among kinds and among concentrations of growth regulators. The optimum cutting time was April to May in hardwood cutting. The application of IBA 1000 $mgL^{-1}$(rooting rate : 36.4%) was most effective in callus formation and rooting of cutting. Relatively, rooting of cutting of the control taken in May was 30.4%. Root characteristics such as number, length and diameter of root were not significantly affected by kinds and concentrations of growth regulators in hardwood cutting.

Selection of Suitable Varieties of Carnation (Dianthus caryophyllus L.) and Optimization of Culture Conditions for Efficient Tissue Culture (효율적 조직배양체계 확립을 위한 카네이션 품종 선발 및 배양조건 설정)

  • Kang, Chan-Ho;Han, Bum-So;Han, So-Gon;Kown, Sung-Hwan;Song, Young-Ju
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.121-129
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    • 2011
  • As the molecular breeding was progressed, many plant transformation techniques were attained for improving transformation accuracy and used to produce useful transgenic plants. Day by day, new varieties were developed so new transformation techniques required for these newly developed varieties. Carnation (Dianthus caryophyllus L.) is a popular and economically important ornamental plant, all over the world. Keeping this in view, we selected 18 varieties of D. caryophyllus L. commonly available in the market and did optimization of culture conditions for more efficient tissue culture and to get higher number of plants via micro-propagation. Four varieties namely Yellowdotcom, Jakarta, Belmonte, Polartessino etc. were selected for organ culture studies from single cell line. The optimum growth was recorded in the MS media supplied with sucrose 3%, NAA 1.0 mg/L and TDZ 1.0 mg/L. except Belmonte, in which, BA 1.0 mg/L was found to be the best combination, in place of TDZ, rest ingredients were same. The most efficient coagulating agent used to obtain higher number of plant from callus was phytagel 0.3%. The most effective explant for higher shoot formation was stem in which 80.2% shoot formation was recorded. It also reduced culture periods by 6 days.

Selection of Herbicide Resistant Potatoes Transformed with Phosphinothricin Acetyltransferase Gene (Phosphinothricin Acetyltransferase 유전자 도입에 의한 제초제 저항성 감자의 선발)

  • Han, S.S.;Jeong, J.H.;Bang, K.S.;Yang, D.C.
    • Korean Journal of Weed Science
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    • v.17 no.4
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    • pp.390-399
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    • 1997
  • This experiment was conducted to introduce PAT (phosphinothricin acetyltransferase, non-selective herbicide bialaphos resistant gene) gene into potato (Solanum tuberosum. cv. Desiree). Optimal shoot regeneration from leaf discs and stem segments was obtained in MS medium supplemented with 0.1 mg/L IBA and 0.5 mg/L BA, and the frequency of shoot regeneration was 54% in left discs and 46% in stem segments. In this condition, leaf discs and stem segments of potato were co-cultivated with A. tumefaciens MP90 which contained binary vector with GUS: :NPTII gene and PAT gene. Transgenic shoots were regenerated from leaf and stem-derived calli on selection medium with 100mg/L kanamycin. The 100${\mu}M$ acetosyringone treatment during the co-cultivation highly enhanced(4 times than the control) the shoot regeneration on selection medium. When the putative transgenic plants were transferred to medium with 10mg/L basta, all of them were survived. After PCR. GUS test, and Southern blot analysis of the survived plant, we confirmed that the gene was stably integrated into the potato genome and expressed. After the transgenic plants were transplanted in soil, and the transgenic plants were sprayed with the herbicide basta (300ml/10a), the transgenic plants remained green but control plants were died.

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Shoot Primordium Culture for Multiplication of Carrot (당근의 다량증식을 위한 순원기 배양)

  • 서호범;이수성
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.93-97
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    • 1999
  • Shoot tips with 2 leaf primordia were cultured to induce shoot primordia in MS liquid medium supplemented with several concentrations of BA and hIAA under the conditions of 10,000 lux illuminations for 24 h and of vertical shaking of 2 rpm in carrot. Two F$_1$ hybrids and two male sterility lines were used. Shoot primordia were only induced in the medium supplemented with 2.0 mg/L of BA and 0.2 mg/L of NAA. Genotypic specificity and seasonal effect of donor parents on shoot primordia induction were not observed and average 15-20% of the planted dornes developed to shoot primordia. The induced shoot primordia were successfully propagated by subculture in the same medium. However, they were grown into three different types during multiplication, that is, the type with multiple small shoots on the surface, the type of without any shoot, and the type of callus. Shoot primordia clusters with small shoots on the surface differentiated multiple shoots successfully in 1/2 MS solid medium supplemented with 0.2 to 1.0 mg/L of IAA and 0.2 to 1.0 mg/L of kinetin. New shoot primordia with small shoots were well formed when pieces bigger than 2 mm in diameter of the out layer of the shoot primordia cluster with small shoots were subcultured. No differences of multiplication and shooting ability and chromosomal variation of shoot primordia were observed until the 13th sub-culture.

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The Filter Membrane Culture Procedure with Feeder Cells in Rice Protoplast Culture (Filter membrane과 feeder세포를 이용한 벼의 원형질체 배양)

  • LEE, Sung-Ho;SHON, Young Geol;Lee, Soo In;DAVEY Micheal R.;COCKING Edward C.;CHO, Moo Je
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.295-303
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    • 1997
  • To investigate the response on feeder cell cultures, protoplasts isolated from cell suspensions initiated from mature seed scutellum-derived callus of the Japonica rice variety Taipei 309, were cultured on filter membranes under various conditions. The effects of various factors, such as gelling agents, feeder cell and protoplast densities, species of feeder cells and heat shock treatment, have been investigated to improve protoplast plating efficiencies on filter membranes. Maximum protoplast plating efficiencies were obtained when protoplasts were cultured on KPR medium semi-solidified with Sea Plaque agarose at a density of $5\;\times\;10^{5}\;ml^{-1}$ protoplasts in the presence of Lolium multiflorum as feeder cells (0.5 ml pcv per 10 ml of protoplast culture medium). Pre-culture heat shock treatments for 1 min. and 5 min. to the protoplasts did not give any appreciable increase on the plating efficiency of protoplasts in the presence of feeder cells. Maltose-supplemented medium was superior for plant regeneration from protoplast-derived colonies compared with medium containing only sucrose. The plants were transferred to the glasshouse, flowered and were fertile.

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A Study on the Effect of Mountain Ginseng Adventitious Roots Extract (산삼부정근 추출물의 효능${\cdot}$효과에 관한 연구)

  • Yoo Yung-Geun;Joung Min-Seok;Lee Youn-Hee;Choi Jong-Wan;Kim Joong-Hoi;Paek Kee-Yoeup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.377-383
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    • 2004
  • This study reviewed the application of an extract from mountain ginseng adventitious roots which had been grown through tissue culture as a cosmetic ingredient. The mountain ginseng adventitious roots were derived from mountain ginseng callus that was induced from mountain ginseng root whose origin is estimated to date back about one hundred years ago. The adventitious roots were separated from callus and grown in a 20 L bioreactor. In order to proliferate the adventitious roots, they were cultured for 5 weeks in bioreactor. Then the harvested mountain ginseng adventitious roots were dried and extracted. For verifying skin whitening effect of an extract from the tissue-cultured mountain ginseng adventitious roots in vivo, we performed the clinical test of it. The research showed the significant skin whitening effect of a mountain ginseng adventitious roots extract and the statistical analysis showed a significant difference (p<0.0001) between sample ($2\%$ mountain ginseng adventitious roots extract) and placebo. But, some saponins showed below $10\%$ inhibitory effect of tyrosinase and melanin synthesis in B-16 melanoma. The extracts of red ginseng and ginseng which were the same concentration as the tissue-cultured mountain ginseng adventitious roots extract's showed little inhibitory effect of tyrosinase and melanin synthesis in B-16 melanoma. In DPPH test, Anti-hydroxyl radical activity of $0.5\%$ the tissue-cultured mountain ginseng adventitious roots extract was $86\%.$.

The Study on Tissue-Cultured Echinacea purpurea Adventitious Roots Extract for Application as a Cosmetic Ingredient (조직 배양한 에키네시아 추출물에 관한 효능 연구)

  • Park, Chang-Min;Joung, Min-Seok;Choi, Jong-Wan;Paek, Kee-Yoeup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.2
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    • pp.137-142
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    • 2008
  • Echinacea purpurea, an indian traditional plant medicine has been widely used as herbal remedy for the treatment of disease such as colds or other infections. However, Echinacea purpurea extracts recently have been applied as a cosmetic ingredient for skin care. We artificially cultured Echinacea purpurea by using the bioreactor culture system for this study. We induced callus from Echinacea purpurea and separated adventitious roots, harvested and extracted after cultured in bioreactors. Previously, several studies have been reported on anti-oxidant and immuno-enhancing effects of Echinacea purpurea extract but other efficacies were not well known. In this study, we investigated the whitening, anti-wrinkle and anti-oxidant effects to know applicable value of tissue-cultured Echinacea purpurea adventitious roots extract(TCEPARE) as a cosmetic ingredient. TCEPARE did not show cytotoxicity until a concentration of 2% and showed the anti-oxidative effect in DPPH and NBT tests. Also, the extract decreased tyrosinase expression in a dose-dependent manner and inhibited melanin synthesis in B16 melanoma cells. TCEPARE reduced protein levels of MMP-1, 2 secreted in culture medium or in cell lysates. From these results we suggest that TCEPARE has potential benefits applicable as to cosmetic ingredient for skin care products.

산삼의 배양 및 그 응용에 관한 연구

  • 신미희
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.2
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    • pp.45-56
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

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