• Title/Summary/Keyword: plant suspension cultures

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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.

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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Effect of Sugar Starvation on the Sugar Transport System in Suspension Cultures of Streptanthus trotus (Streptanthus tortus 배양 세포에서 당류고갈이 당류 수송계에 미치는 영향)

  • 조봉희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.47-50
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    • 2000
  • In suspension cultures of Streptanthus. the uptake rate of sugar was increased during the ceil starvation of sugar in the medium. The maximal uptake rate obtained with 3 days of cell starvation. Sugar transport system induced by the sugar starvation was completely inhibited by 10 $\mu$M cycloheximide. Plant cells are known to possess only one sugar transport system, but the uptake rate of glucose obtained a saturated kinetic while the one of sucrose had two different kinetics after the sugar starvation. Induced sugar transport systems had different kinetics compared to plant cell. These results showed that higher plants have adaptable ability to induce new sugar transport systems when the environment changed unsuitable.

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Production of miraculin protein in suspension cell lines of transgenic rice using Agrobacterium (Agrobacterium을 이용한 형질전환 벼 현탁 세포주에서 miraculin 단백질의 생산)

  • Kim, Hee Kyoung;Go, Ji Yun;Park, So-Young;Kang, Kwon Kyoo;Jung, Yu Jin
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.227-234
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    • 2020
  • To produce the miraculin protein in suspension cultures, rice (Oryza sativa L.) was transformed with Agrobacterium tumefacience EHA105 containing the miraculin AB512278 gene. The cell suspension cultures were established using cell lines selected from transgenic rice callus. The integration of the miraculin gene into the rice chromosome was confirmed using genomic PCR analysis. In addition, RT-PCR analysis indicated that the miraculin gene is expressed in the selected suspension cell lines. Thus, the recombinant miraculin was expressed in the transgenic suspension cell line, HK-2. Therefore, we have successfully developed a HK-2 line that produces miraculin. These results demonstrate that transformed cell suspension cultures can be used to produce a taste-modifying protein such as miraculin.

Increased production of human granulocyte-macrophage colony stimulating factor (hGM -CSF) by the addition of stabilizing polymer in plant suspension cultures

  • Kim, Nan-Seon;Lee, Jae-Hwa;Kim, Yeong-Suk;Gwon, Tae-Ho;Yang, Mun-Sik
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.95-98
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    • 2001
  • The effect of stabilizing polymer on hGM-CSF production was investigated in suspension cell cultures of transgenic tobacco. Secreted human GM -CSF from cell suspension cultures was detected in the medium at a maximum concentration of 180 ${\mu}g/L$ by ELISA. However, the secreted hGM -CSF was unstable in the medium, and rapidly degraded after day 5. In order to stabilize the secreted hGM-CSF, three stabilizing polymers were tested, polyethylene glycol, polyvinylpyrrolidone and gelatin. Gelatin was the most effective in stabilizing the secreted GM-CSF. Following the addition of 5% (w/v) gelatin, the maximum GM -CSF concentration reached 783 ${\mu}g/L$, a 4.6-fold increase over control.

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Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Production of Rosmarinic Acid, Lithospermic Acid B, and Tanshinones by Suspension Cultures of Ti-Transformed Salvia miltiorrhiza Cells in Bioreactors

  • Zhong, Jian-Jiang;Hui Chen;Feng Chen
    • Journal of Plant Biotechnology
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    • v.3 no.2
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    • pp.107-112
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    • 2001
  • The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

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Purification of Phospholipase $A_2$ from Scutellaria baicalensis Suspension Cells (황금 배양 세포로부터 Phospholipase $A_2$의 분리)

  • Ma, Choong-Je;Kim, Dae-Kyung
    • Korean Journal of Pharmacognosy
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    • v.40 no.1
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    • pp.13-17
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    • 2009
  • It was previously reported that yeast elicitor transiently increased oleanolic acid and ursolic acid in Scutellaria baicalensis suspension cultures and also doubled phospholipase $A_2$ ($PLA_2$) activity. Thus, $PLA_2$ was purified from the soluble fractions of S. baicalensis suspension cultures and the characters of the purified $PLA_2$ were identified. The $PLA_2$ was purified about 160 times compared with the starting soluble-protein extract from S. baicalensis suspension culture cells. The purified protein showed a molecular mass of about 43 kDa by SDS-PAGE. The purified plant $PLA_2$ had a neutral pH optimum (pH 7.0) and required $Ca^{2+}$ for activity. The $PLA_2$ activity was inhibited by mammalian $PLA_2$ inhibitors such as 5,8,11,14-eicosatetraynoic acid(ETYA) and arachidonyl trifluoromethyl ketone ($AACOCF_3$).

Effect of Inoculum Size on Biomass Accumulation and Ginsenoside Production by Large-Scale Cell Suspension Cultures of Panax ginseng

  • Thanh Nguyen Trung;Murthy Hosakatte Niranjana;Yu Kee-Won;Jeong Cheol Seung;Hahn Eun-Joo;Paek Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.265-268
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    • 2004
  • Cell growth and ginseng saponin production by large-scale suspension (bioreactor) cultures of Panax ginseng were investigated under various inoculum sizes. Cell growth was low at an inoculum size of 40 g FW/L, and the maximum cell growth was obtained with increasing inoculum size up to 100 g FW/L. The cell density of 333 g FW/L and 12.7 g DW/L was obtained at inoculum size of 100 g FW/L after 30 days of cultivation. Maximum saponin production of $4.40\;\cal{mg/g}$ DW was achieved at 60 g FW/L of inoculum size. Thus, inoculum size 60 g FW/L was suitable for optimum biomass accumulation as well as saponin production during bioreactor cultivation of ginseng suspension cells.

Selection of Cell Source and the Effect of pH and MS Macronutrients on Biomass Production in Cell Cultures of Tongkat Ali (Eurycoma longifolia Jack)

  • Siregar Luthfi Aziz Mahmud;Chan Lai-Keng;Boey Peng-Lim
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.131-135
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    • 2003
  • Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.