• Title/Summary/Keyword: Cell suspension

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Elicitor-induced accumulation of stilbenes in cell suspension cultures of Cayratia trifolia (L.) Domin

  • Roat, Chetana;Ramawat, K.G.
    • Plant Biotechnology Reports
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    • v.3 no.2
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    • pp.135-138
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    • 2009
  • Cell cultures of Cayratia trifolia (Vitaceae), a tropical lianas, were maintained in Murashige and Skoog's medium containing $0.25mg\;1^{-1}$ NAA, $0.2mg\;1^{-1}$ kinetin and casein hydrolysate $250mg\;1^{-1}$. Cell suspension cultures of C. trifolia accumulate stilbenes (piceid, resveratrol, viniferin, ampelopsin), which on elicitation by any of $500{\mu}M$ salicylic acid, $100{\mu}M$ methyl jasmonate, $500{\mu}M$ ethrel and $500mg\;1^{-1}$ yeast extract, added on the 7th day, were enhanced by 3- to 6-fold ($5-11mg\;1^{-1}$) by the 15th day.

DESIGN OF A CHECK VALVE FOR FEEDING BIOLOGICAL CELLS ONE BY ONE (세포의 개별 공급을 위한 체크 밸브의 설계)

  • Choi, Jung-Hoon;Lee, Sang-Wook;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 1994.07a
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    • pp.85-87
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    • 1994
  • Feeding biological cells one by one is the key point in the manipulation of cells. The conventional valve systems have many difficulties in feeding cells one by one, because they shut the whole flow of fluids when they are closed and have possibilities of breaking the fragile cells. They need some other equipments for continuous supply of suspension and to protect the cells. We design a check valve for feeding biological cells one by one using polyimide all the silicon substrate. The cells are fed by hydraulic pressure through the isotropically etched cavity. When the suspension flows continuously along the channel the valve is bent by hydraulic pressure and a cell is fed to the outlet. We have studied a cell fusion device fabricated with polyimide and electroplating. If the designed check valve is located in front of the cell fusion device it is helpful to fuse two different kinds of cells.

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Effects of Various Elicitors on the Production of Berberine in Plant Cell Suspension Cultures of Thalicrtrum rugosum (Thalicrtrum rugosum 세포배양에 의한 berberine 생산에 미치는 여러 가지 elicitor의 영향)

  • 윤정환;박인석김동일
    • KSBB Journal
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    • v.8 no.4
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    • pp.390-396
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    • 1993
  • Effects of various elictitors were investigated to enhance the production of berberine in plant cell suspension cultures of Thalicrtrum rugosum. Treatments of yeast elicitor, 15 different types of abiotic elicitors, 16 kinds of fungal elicitors from three species of fungi were performed. Cell growth and berberine production were examined and compared for both normal and elicitor treated cultures. No distinguished increases in berberine yield by the addition of elicitors could be attained.

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Studies on the Induction of Transformation and Multiplication in Orchid Plants.(III) Expression of Gene Transferred into Orchid Protoplasts by Electroporation (난과식물의 형질전환 유도 및 다량증식에 관한 연구. III. Electroporation에 의해서 자란의 원형질체로 도입된 유전자의 발현)

  • 이정석;황성진김영준황백
    • KSBB Journal
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    • v.6 no.4
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    • pp.385-388
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    • 1991
  • Embryogenic cell suspension cultures of B. striata were established as transferred selected embryogenic callus into liquid medium. Protoplasts isolated from embryogenic cell suspensions were electroporated in buffered solutions containing plasmid DNA of pBI121. Transient GUS (beta-glucuronidase) activity measurement and selection for kanamycin resistent showed that expression of foreign genes and stable transformation were achieved. GUS transient gene expression was increased by increasing DNA concentration of pBI121 plasmid and affected by the level of the applied voltage. An optimal level of GUS activity was obtained after electroporation with a pulse of 200-300 voltage/1180 uF. Protoplast viability was up to the 80% at the optimal voltage.

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Isolation and Culture of Protoplasts Derived from Embryogenic Cell Suspension Culture of Oryza sativa (Rice) (벼 진탕 배 배양세포로부터 원형질체 분리 및 배양)

  • Hwang, Baik;Kim, Mee-Kyung;Vasil, I. K.
    • Journal of Plant Biology
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    • v.31 no.1
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    • pp.41-49
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    • 1988
  • Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2, 4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2, 4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.

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Long-Term Starvation Induces the Viable-but-Nonculturable Condition in Lactobacillus crispatus KLB46

  • Lee, Seok-Yong;Kim, Ju-Hyeon;Jang, Jeong-Eun;Kim, Seung-Cheol;Yun, Hyeon-Sik;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.918-922
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    • 2001
  • In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

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Production of Immunostimulating Polysaccharide in Angelica gigas Nakai SusDension Cell Cultures (참당귀 현탁세포배양에 의한 면역증강성 다당 생산)

  • Kim, Young-Hwa;Kim, Ik-Hwan;Kim, Dong-Il
    • KSBB Journal
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    • v.21 no.5
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    • pp.331-335
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    • 2006
  • Suspension cells of Angelica gigas Nakai were cultivated to produce extracellular polysaccharide(ECP) as immunostimulating agents. Effects of environmental conditions such as sucrose and 2,4-dichlorophenoxyacetic acid(2,4-D) concentrations on the growth and production of ECP were studied using suspension cultures of A. gigas Nakai. Final dry cell weight was increased with an increase of initial sucrose concentration from 30 to 60 g/L. The maximum production of ECP(1.2 g/L) was achieved at an initial sucrose concentration of 50 g/L on day 8. High 2,4-D concentration was effective for ECP production but not for cell growth. In addition, various fungal elicitors were investigated for the enhanced production of ECP in A. gigas suspension cultures. Among the tested fungal elicitors, Verticillium dahliae was the most effective for the production of ECP in A. gigas suspension culture.

Identification of Salidroside from Rhodiola sachalinensis A. Bor. and its Production through Cell Suspension Culture (참돌꽃에서 Salidroside의 동정 및 현탁세포배양을 통한 분리)

  • Kim, Soo-Jung;Kim, Kwang-Soo;Hwang, Sung-Jin;Chon, Sang-Uk;Kim, Young-Ho;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.203-208
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    • 2004
  • Salidroside was isolated and purified from R. sachalinensis A. Bor. roots. Purified salidroside was obtained from repeated silicagel column chromatography and preparative HPLC, and identified by $^1H-NMR,\;^{13}C-NMR\;and\;^1H-^1H$ COSY spectra analyzer. Callus induction and cell suspension from R. sachalinensis leaf segments were established on 1/2MS solid medium and in $2B_5$ liquid medium containing 0.5 mg/l NAA and 1 mg/l, BA in the dark condition, respectively. The contents of salidroside for suspension culture were ranging from 0.12% to 0.41% in comparison with 0.17% for natural roots.

Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.125-131
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    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

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Embryogenic cell suspension culture and plant regeneration in zoysiagrass (Zoysia japonica Steud) (한국들잔디 배아세포의 부유배양과 식물체 재생)

  • Fang, Wenjuan;Han, Liebao;Qi, Chunhui;Li, Deying;Park, Tae-Yun
    • Asian Journal of Turfgrass Science
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    • v.23 no.2
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    • pp.345-352
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    • 2009
  • Zoysiagrass (Zoysia japonica Steud) is a warm season turfgrass species widely used for sports field and golf courses. Many cultivars are propagated through vegetative methods. This study was conducted to develop an optimum culture medium and culture conditions for embryogenic callus induction and plant regeneration, and to establish a cell suspension culture system for use in zoysiagrass breeding and propagation. The results indicated that adding $Cu^{++}$ at 2.5 mg $L^{-1}$ to the induction medium was optimum for callus induction. Increasing the numbers of sub-culture cycles improved the quality of calli. The optimum dosage for cell suspension culture ranged from 2.5 to 10 mL. The embryogenic callus suspension used in this study had a plant regeneration rate of 58%.