• Title/Summary/Keyword: plant cultured cells

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Glutathione Contents in Various Plant Cell Lines (다양한 식물배양세포주의 Glutathione 함량)

  • 이정은;안영옥;권석윤;이행순;김석원;박일현;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.57-61
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    • 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.

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Development of natural sunscreen using plant extracts (식물 추출물을 이용한 천연 자외선 차단제 개발)

  • Moon, Ji-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.5
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    • pp.1138-1150
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    • 2020
  • In this study, an experiment was conducted to develop a sunscreen with antioxidant effects by simultaneously investigating the antioxidant and UV protection capabilities of various plant extracts. First, to investigate the UV-blocking ability of 33 kinds of plant extracts, the absorbance spectrum between the UV wavelength of 280 to 400 nm was investigated. Arrowroot, graviola, wheat sprout, sangbaek skin, thorn meal, lacquer, etc. 11 species were selected. The total polyphenol content, total flavonoid content, and DPPH radical scavenging activity of the selected plant extracts are measured to examine the degree of antioxidant activity, and from this, it is a plant extract that has excellent UV protection and antioxidant activity at the same time. The species was selected. A gel-shaped cream is prepared by mixing the selected gold, hops, and licorice extracts in a ratio of 1:1:1, and the UV protection effect of this cream is measured when the cultured cells are irradiated with UV rays. Determined by the method. As a result of the study, it was confirmed that the selected mixture of plant extracts complemented each other in terms of ultraviolet absorption ability and increased cell damage protection effect. Through these results, it was confirmed that it was possible to develop a sunscreen with an antioxidant effect if the antioxidant and sunscreen capabilities of various plant extracts were determined at the same time.

Development of Transgenic Plant (Codonopsis lanceolata Trautv.) Harboring a Bialaphos Resistance Gene, bar (Bialaphos 저항성 유전자 bar를 이용한 형질전환 더덕개발)

  • 조광수;장정은;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.281-287
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    • 1999
  • Codonopsis lanceolata ("Deoduck" in Korea) is a perennial herb, and belongs to family, Campanulaceae. Its taproot is used a good source of a wild vegetable as well as an herbaceous medicine. In this study, to develop a bialaphos-resistant transgenic Codonopsis, seed germination mechanism and somatic embryogenesis of the plant were investigated, and Agrobacterium-mediated transformation with bar gene encoding phosphinothricin acetyltransferase (PAT) was performed. Attempt were made to regenerate plant from cells via somatic embryogenesis. When the cotyledons, nodes and leaf disks were cultured on MS medium containing 2,4-D and zeatin, embryogenic calli were induced. Upon transferring the somatic embryos to N6 solid medium without plant growth regulators, they developed into plantlets under continuous illumination. All plants were dead on MS basal medium containing 10 mg/L phosphinothricin (PPT) and Basta, respectively. The explants did not produce calli in the medium containing 200 mg/L kanamycin. The explants were cocultured with Agrobacterium tumefaciens for 2 days, and transformants were selected in MS basal medium containing 1.0 mg/L 2,4-D, 100 mg/L kanamycin and 500 mg/L carbenicillin. After the selection, embryogenic calli were induced and then somatic embryos were produced by subsequent subculturing. The somatic embryos were germiated on N6 basal medium containing 200 mg/L kanamycin and 500 mg/L carbenicillin. PCR analysis showed that nptII and bar genes were introduced in the Deoduck transformants. After the confirmation of bar gene expression in RNA and protein level, the transgenic Deoduck will be used to study the genetics of filial generation with the herbicide control gene, bar.gene, bar.

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Transgenic Tomato Plants That Overexpress Superoxide Dismutase in Fruits (토마토 과실에서 Superoxide Dismutase를 고발현하는 형질전환 식물체)

  • Park, Eun-Jeong;Lee, Haeng-Soon;Kwon, Suk-Yoon;Choi, Kwan-Sam;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.7-13
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    • 2002
  • Superoxide dismutase (SOD) plays an important role in cellular defense against oxidative stress in plants. We have developed transgenic tomato plants overexpressing a cassava SOD in fruits. Three transgenic tomato plants (one from cv. Pink forcer and two from cv. Koko) using a new vector system, ASOp :: . mSOD1/pBI101, harboring ascorbate oxidase promoter (ASOp) expressing dominantly in cucumber fruits, CuZnSOD cDNA (mSOD1) isolated from cultured cells of cassava, and nptll gene as a selectable marker were successfully developed. SOD specific activity (units/mg protein) in transgenic fruits of both cultivars was increased with maturation of the fruits. SOD specific activity of well-mature fruits in transgenic Pink forcer and Koko showed approximately 1.6 and 2.2 times higher than control fruits, respectively. The strength of SOD isoenzyme bands well reflected the SOD activity during the fruit maturation. These results suggested that SOD gene was properly introduced into tomato fruits in a fruit-dominant expression manner by ASO promoter.

Extract of high hydrostatic pressure-treated danshen (Salvia miltiorrhiza) ameliorates atherosclerosis via autophagy induction

  • Ko, Minjeong;Oh, Goo Taeg;Park, Jiyong;Kwon, Ho Jeong
    • BMB Reports
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    • v.53 no.12
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    • pp.652-657
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    • 2020
  • Danshen (Salvia miltiorrhiza) is a traditional medicinal plant widely used in Asian countries for its pharmacological activities (e.g., amelioration of cardiovascular diseases). In this study, we investigated the anti-atherosclerotic activity of raw danshen root extract prepared using high hydrostatic pressure (HHP) at 550 MPa for 5 min and hot water extraction. This method was useful for elimination of bacteria from cultured danshen plants and for better extraction yield of active principles. The HHP-treated danshen extract (HDE) inhibited proliferation of human umbilical vein endothelial cells (HUVECs) and induced autophagy that was assessed by LC3 conversion and p62 degradation. HDE suppressed foam cell formation in oxLDL-induced RAW264.7 macrophages; lysosomal activity simultaneously increased, measured by acridine orange staining. HDE also reduced atherosclerotic plaque development in vivo in apolipoprotein E knock-out (ApoE-/-) mice fed a high cholesterol diet. Taken together, these results indicated that HDE exhibited anti-atherosclerotic activity via autophagy induction.

In vitro regeneration and the change of anatomical appearance in Poncirus trifoliata RAFIN. (탱자(Poncirus trifoliata RAFIN.)의 기내 재분화 및 조직학적 특성)

  • 박민희;이현화;장현규;이숙영;김홍섭
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.107-119
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    • 1999
  • In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

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Effects of Phytoestrogen on Cell Growth and Insulin-like Growth Factor-I (IGF-I) Production in MC3T3-El Cells (식물성 에스트로겐이 MC3T3-El 골아세포의 성장과 Insulin-like Growth Factor-1(IGF-1)생성에 미치는 영향)

  • Kwon, Ji-Young;Nam, Taek-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.743-749
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    • 2005
  • Estrogen is known to play an important role in maintaining bone mass, since the concentration of serum estrogen decrease after menopause and the estrogen deficiency results in bone loss. Phytoestrogens are plant compounds with estrogen-like biological activity, In this study, to investigate the bioactivities of phytoestrogen, which act on bone metabolism, we examined the effect of selected food-borne phytoestrogens (genistein, daidzein and resveratrol) on osteoblast proliferation and IGF-I production using MC3T3-El cells, a mouse calvaria osteoblast-like cell line. Cells were cultured in a serum free medium for 48 hr in the presence of genistein $(10^{-5}\;M)$, daidzein $(10^{-5}\;M)$ and resveratrol $(10^{-5}\;M)$. The effects of genistein, daidzein and resveratrol on the cell proliferation and growth were evaluated by total cell numbers, MTS assay and cell migration assay. Their effect was compared with the $17\beta-estradiol$. Genistein, daidzein and resveratrol exhibited stimulatory effects on the growth of MC3T3-El cells, and the most pronounced effect was shown with daidzein. In addition, these phytoestrogen increased alkaline phosphatase activity of MC3T3-El cells. These effects were similar to that of $17\beta-estradiol$ effects. Moreover, treatment with genistein, daidzein and resveratrol increased production of insulin like growth factor-I (IGF-I) in conditioned media, indicating that the growth promoting effects of these phytoestrogen were related to the changes in production of IGF-I by MC3T3-El cells. These results show that genistein, daidzein and resveratrol have a stimulatory effect on osteoblast function, and that these findings in a cell model may prove relevant to protecting against the loss of bone mass and the development of osteoporosis in human subjects.

Development of an automatic system for cultivating the bioluminescent heterotrophic dinoflagellate Noctiluca scintillans on a 100-liter scale

  • You, Ji Hyun;Jeong, Hae Jin;Park, Sang Ah;Ok, Jin Hee;Kang, Hee Chang;Eom, Se Hee;Lim, An Suk
    • ALGAE
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    • v.37 no.2
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    • pp.149-161
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    • 2022
  • Noctiluca scintillans is a heterotrophic dinoflagellate that causes red-colored oceans during the day (red tides) and glowing oceans at night (bioluminescence). This species feeds on diverse prey, including phytoplankton, heterotrophic protists, and eggs of metazoans. Thus, many scientists have conducted studies on the ecophysiology of this species. It is easy to cultivate N. scintillans at a scale of <1 L, but it is difficult to cultivate them at a scale of >100 L because N. scintillans cells usually stay near the surface, while prey cells stay below the surface in large water tanks. To obtain mass-cultured N. scintillans cells, we developed an automatic system for cultivating N. scintillans on a scale of 100 L. The system consisted of four tanks containing fresh nutrients, the chlorophyte Dunaliella salina as prey, N. scintillans for growth, and N. scintillans for storage, respectively. The light intensities supporting the high growth rates of D. salina and N. scintillans were 300 and 20 µmol photons m-2 s-1, respectively. Twenty liters of D. salina culture from the prey culture tank were transferred to the predator culture tank, and subsequently 20 L of nutrients from the nutrient tank were transferred to the prey culture tank every 2 d. When the volume of N. scintillans in the predator culture tank reached 90 L 6 d later, 70 L of the culture were transferred to the predator storage tank. To prevent N. scintillans cells from being separated from D. salina cells in the predator culture tank, the culture was mixed using an air pump, a sparger, and a stirrer. The highest abundance of N. scintillans in the predator culture tank was 45 cells mL-1, which was more than twice the highest abundance when this dinoflagellate was cultivated manually. This automatic system supplies 100 L of N. scintillans pure culture with a high density every 10 d for diverse experiments on N. scintillans.

Effect of n-3 Fatty Acids on Estrogen Dependency and Protein Kinase C Activity of Human Breast Cancer Cell

  • Cho, Sung-Hee;Oh, Sun-Hee;Park, Hee-Sung
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.220-226
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    • 1996
  • To investigate the effect of long chain n-3 polyunsaturated fatty acids on breast cancer cell growth, estrogen-dependent MCF-7 human breast cancer cells were cultured serum-free DMEM media containing 0.5$\mu\textrm{g}$/ml of differnet kinds of fatty acids; linoleic acid(LA), arachidonic acid(AA), eicosapentaenoic acid(EPA) and docosahexaenoic acid acid(DHA) and 1, 0.1, 0.2, 0.5and 1.0ng/ml 17$\beta$-estradiol as well as 10$\mu\textrm{g}$/mi insulin and 1.25 mg/ml delipidized bovine serum albumin for 3 days. Cell growth monitored by MTT assay was lower in DHA and EPA treatments as compared to LA treatment, but not with AA treatment. Estrogen concentrations at which cell growth was initially stimulated were 0.1ng/ml for LA and DHA treatments and 0.2ng/ml for EPA and AA treatments, but the degree of stimulation was 25~30% lower in DHA and EPA treatments than in LA treatment. Fatty acid analysis showed that each fatty acid in culture medium was well incoporated into celluar lipid. Protein kinase C activity of cells was most elevated in LA treatment from 2 to 8 hours of culture followed by DHA, EPA, and AA treatments. It is concluded that inhibitions of n-3 DHA and EPA on breast cancer cell growth as compard with n-6 LA is mediated via changes in membrane fatty acid composition reducing estrogen sensitivity and increasing protein kinase C activity.

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Anthocyanin Synthesis in Cell Cultures of Populus alba L. × P. glandulosa Uyeki (세포배양(細胞培養)을 이용(利用)한 현사시나무의 안토시아닌 생성(生成))

  • Park, Young Goo;Choi, Myung Suk;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.81 no.2
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    • pp.183-190
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    • 1992
  • The influence of various levels of major medium components such as sucrose, nitrate, phosphate, plant growth regulators, and light intensity for cell growth and the production of anthocyanin content in cell cultures of Populus alba ${\times}$ P. glandulosa were investigated. Best results for anthocyanin yield were obtained using Murashige and Skoog(MS) medium containing 5% sucrose, 12.5% nitrate, 200% phosphate, 1.0mg/l indole-3-acetic acid(IAA), 1.0mg/l benaylaminopurine(BAP), and continuous illumination of 7,000 lux. On the other hand, maximum cell growth was achieved with 5% sucrose, 50% nitrate above 400% phosphate compare with that of MS basal mediumi, and 0.5mg/l 2, 4-dichlorophenoxyacetic acid(2, 4-D). Anthocyanin accumulation in a suspension cultured cells of given genotype was stimulated by subculturing onto the medium lacking 2, 4-D. Pigmented cell clusters were extracted with methanol containing 1% hydrochloric acid (HCl) and then anthocyanin was identified by thin layer chromatography (TLC) and U. V. spectrophotometer.

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