• Title/Summary/Keyword: Suspension Culture

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Suspension Culture of Gardenia jasminoides Ellis Cell for Production of Yellow Pigment

  • Kim, Sang-Hwa;Park, Young-Goo;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.142-149
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    • 1991
  • Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

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Elicitation of Seedlings and Cultured Cells for the Production of Capsidiol in Capsicum annum L. (고추 (Capsicum annum L.)식물체 및 배양세포의 Capsidiol 생산 유도)

  • 권순태;정은아;박해영;손건호
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.249-254
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    • 2001
  • Effects of ultraviolet stress and elicitors, cellulase and jasmonic acid (JA), for the production of capsidiol, sesquiterpenoid phytoalexin, in seedlings and suspension cultures of pepper (Capsicum annum L. cv, Soobicho) were examined. Extracellular capsidiol in the medium of suspension cultures was absent from control cells, but accumulated in the elicitor treated cells with 0.05 $\mu\textrm{g}$/mL of cellulase or 0.1 $\mu\textrm{g}$/mL JA. Elicited cells gradually decreased their viability and eventually died within 48 hours of elicitor treatment by the toxicity of capsidiol accumulated in the culture medium. Capsidiol production in the leaves of pepper seedlings was markedly increased by the treatment of ultraviolet stress and reached maximum level at 48 hours of irradiation. Infiltration of elicitors, 0.05 $\mu\textrm{g}$/mL cellulase or 1.0 $\mu\textrm{g}$/mL JA, to the surface of leaf or fruit, stimulated the elicitation of the cells which resulted in the production of capsidiol and expansion of pathogene-like lesion around the elicitor treated region.

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Callus Induction and Somatic Embryogenesis in Suspension Culture of Rehmannia glutinosa (지황의 캘러스 유도와 현탁배양에서 체세포배 발생)

  • Chae, Young-Am;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.184-190
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    • 1993
  • This study was carried out to investigate the appropriate medium and constitutionof growth regulators for somatic embryogenesis for development of rapid mass propagation system via somatic embrygenesis in Rehmennia glutinosa. Embryogenic callus formation from leaf explant was more effective when 4mg / l BA with 0.5mg / l NAA than that of treated with only auxins or cytokinins. LS medium was suitable for embryogenic callus formation. LS medium with 4mg / l BA with 0.5mg / l NAA was effective for the maintenance and proliferation of embryogenic callus. In suspension culture, addition of 1mg / l BA to LS medium was proper for somatic embryogenesis. The highest rate of shoot developement form cotyledon stage embryo was obtained in 1/2 LS medium and plantlet survived by 75% after transplanted to the soil. after 4 weeks.

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The Component Analysis of the Bioflocculant Produced by Strain YG-02 (Strain YG-02가 생산하는 생물응집제의 성분 분석)

  • Jung, Yeon-Gon;Ko, Joon-Il;Chung, Seon-Yong
    • Journal of Korean Society on Water Environment
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    • v.35 no.3
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    • pp.201-208
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    • 2019
  • In this study, we analyzed composition of the bioflocculant, which strain YG-02 produces. First, supernatant and suspension from centrifugation of culture fluid of the strain, were used in the flocculation experiment. As a result, the SVI(sludge volume index) added with the suspension, was 182 mL/g, same as the control group with no additive, and the SVI added with supernatant, was 164 mL/g. So, the result above showed that flocculation capacity of the bioflocculant, was dependent on the substance which strain YG-02 produces, not on factors such as the body of germs. As a result of the thermostability test on substances that cause flocculation, the flocculation effect was significantly reduced, compared to the result of the flocculation test, before applying heat to the culture fluid, and it was able to assume that the substance that causes flocculation, was damaged by heat. Additionally, to understand the component of the bioflocculant, analyzation of sugar composition and fatty acid, was conducted. As a result, sugar composition was the polysaccharide consisting of glucose: lactose with molar ratio of 90.75:9.25. Fatty acid content was detected, as 0.0012 g/100g, showing that it contained glycolipid in the bioflocculant. Such results show that the bioflocculant which strain YG-02 produces, is the new bioflocculant, different from bioflocculantstudiedto date.

Enhanced Production of Shikonin by Using Polyurethane-entrapped Lithospermum erythrorhizon Cells (Polyurethane Foam 에 포괄시킨 Lithospermum erythrorhizon 세포에 의한 Shikonin 생산)

  • Taek, Seo-Weon;Liu, Jang-Ryol;Park, Young-Hoon
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.343-348
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    • 1989
  • Production of shikonin derivatives by Lithospermum erythrorhizon cells by using polyurethane foam was invesliigated. Shikonin derivatives were effectively adsorbed mostly by phase distribution to polyurethane matrices and their production increased significantly compared to the suspension culture. The enhanced production of shikonin was probably due to more facilitated cell to cell con-tact and lowered intracellular shikonin concentration, both of which are known to be favorable for plant secondary metabolite production. In order to improve the process productivity, tell culture was conducted under various culture conditions: Of them, Schenk and Hildebrandt medium containing indole-3-acetic acid (1.75mg/ι) and kinetin (0.1mg/ι) was considered most appropriate for shikonin production. Production of shikonin increased about 4.5 times in the Schenk and Hildebrandt medium containing indole-3-acetic acid (1.15mg/ι) and kinetin (0.1mg/ι) when compared to the same medium containing p-chlorophenoxyacetic acid (2.0mg/ι) and kinetin (0.1mg/ι). When poly-urethane was used as the support material, a single-stage system was more preferred to the conventional two-stage culture system in terms of shikonin productivity.

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Production of Biomass and Bioactive Compounds from Cell Suspension Cultures of Eurycoma longifolia in Balloon Type Bubble Bioreactors

  • Shim, Kyu-Man;Murthy, Hosakatte Niranjana;Park, So-Young;Rusli, Ibrahim;Paek, Kee-Yoeup
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.251-258
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    • 2015
  • Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.

Establishment of a Simple and Rapid Method for Quantitative Determination of -Aminobutyric Acid Using 1H NMR and Production of γ-Aminobutyric Acid in Cell Suspension Cultures of Coriandrum sativum L. (1H NMR에 의한 γ-Aminobutyric Acid의 간단하고 신속한 정량분석법 확립과 고수 (Coriandrum sativum L.) 현탁배양세포로부터 γ-Aminobutyric Acid의 생산)

  • Kim, Suk-Weon;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.207-212
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    • 2007
  • This study describes a simple and rapid method for quantitative determination of $\gamma$-aminobutyric acid (GABA) using $^1H$ NMR spectroscopy from whole cell extracts of plant suspension cultures. When 9 cell lines derived from 8 species of higher plants maintained in liquid Marashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) were subjected to $^1H$ NMR, a cell line of Coriandrum sativum L. exhibited the highest level of GABA. The level reached up to 16.9 mg/dry wt when cells were cultured in MS medium supplemented with 0.5 mg/L 2,4-D after 3 weeks of incubation. The method for quantitative determination of GABA using $^1H$ NMR established in this study could be applied to high-throughput screening of various plant resources for GABA production and the cell suspension culture system of C. sativum could be further developed for commercial production of GABA.

Characteristics of Batch Cultures and Effects of Various Elicitors on Ginsenoside Production in Suspension Cultures of Panax Ginseng C.A. Meyer (고려인삼세포 현탁배양에서 회분배양 특성 및 Ginsenoside 생산에 대한 다양한 elicitors의 영향)

  • 유병삼;변상요
    • KSBB Journal
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    • v.16 no.6
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    • pp.620-625
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    • 2001
  • This study was examined to investigate the time course behaviors of cell growth and sucrose consumption, and effects of various elicitors on ginsenoside production in batch suspension cultures of Panax ginseng Meyer. Suspended cells reached to the stationary phase at 12 days after innoculation. The maximum cell concentration was 14.7 g-DCW/L at 17 days. The highest cell growth rate was 0.59 g-DCW/L d. The sucrose used as a sole carbon source was hydrolysed to glucose and fructose in 4 days, then quickly utilized until the middle-log phase and consumpted completely at 16 days. Various elicitors were app1ied at 8 days from inoculation which is the middle-log phase. Among the elicitors tested, jasmonic acid was the most efficient to increase the ginseneside production, which was 1.5 times higher than control.

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Introduction of Calmodulin into Suspension-Cultured Cells and Protoplasts of Soybean (Glycine max L.) (대두(Glycine max L.) 현탁배양 세포와 원형질체 내로의 외부 Calmodulin의 도입)

  • Hyun Sook CHAE;Kyu Chung HUR;In Sun YOON;Bin G. KANG
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.363-367
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    • 1994
  • In an effort to investigate the role of calmodulin (CaM) as a modulating molecule in the signal transduction system in plant cells, we established methods for introduction of purified CaM into cultured soybean cells. CaM was purified from bovine testis, and was labelled with fluorescein isothiocyanate (FITC). Suspension -cultured cells were healed with saponin (0.1 mg/mL) to permeabilize the plasma membrane and coincubated with FITC-CaM complex. Saponin pretreatment was found to increase the fluorescence in the suspension cultured cells, indicating that the FITC-CaM complex could be incorporated into the cytoplasm. Optimal conditions for introducing FITC-CaM complex into protoplasts by electroporation were established with various electric pulses. With increasing field strength, the fluorescence in the protoplase was increased, while the viability of the protoplase decreased. FITC-CaM complex was successfully introduced into the protoplasts by electroporation and the amount of FITC-CaM complex in the protoplase was estimated.

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Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato (고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성)

  • Park, Soo-Young;Ryu, Sun-Hwa;Kwon, Suk-Yoon;Kim, Jong-Guk;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.