• Title/Summary/Keyword: rice cell suspension culture

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Effects of Silkworm Hemolymph on Cell Viability and hCTLA4Ig Production in Transgenic Rice Cell Suspension Cultures

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Ryu, Hyun-Nam;Yu, Da-Hyun;Choi, Yong-Soo;Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1944-1948
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    • 2007
  • Silkworm hemolymph (SH), prepared from fifth-instar larvae of Bombyx mori and heat-treated at $60^{\circ}C$ for 30 min, was used to improve cell viability and the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic Oryza sativa L. cell suspension cultures. Even though SH could not elevate cell viability at the concentrations up to 3% (v/v), addition of 0.3% (v/v) SH to a culture medium enhanced the production of hCTLA4Ig by 36.8% over an SH-free medium. Moreover, the production period of hCTLA4Ig could be shortened in a 0.3% (v/v) SH-added medium compared with that in an SH-free culture. As a result, addition of 0.3% (v/v) SH improved the productivity of hCTLA4Ig significantly in transgenic rice cell cultures.

Effects of Various Ions on the Cellular and Secretory Isoperoxidases in Rice Suspension Culture

  • Lee, Mi-Young
    • BMB Reports
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    • v.30 no.6
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    • pp.379-384
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    • 1997
  • The effects of several ions on the specific activity and isozyme patterns of cellular and secretory isoperoxidases were studied in suspension-cultured cells of rice (Oryza sativa L.). Peroxidase release into the culture medium occurred in the absence of added calcium. The addition of calcium ion greatly stimulated the secretion of cationic isoperoxidases such as C2 and C3 into the medium: a maximum 11 fold increase of secretions occurred in the presence of 5 mM $CaCl_2$, and the secretion was accomplished within 1 hour after the addition of $CaCl_2$. About a 10 fold increase of the peroxidase secretion into the medium did occur with 0. 5% NaCl, whereas cellular isoperoxidase levels were reduced notably. About a 6 fold increase of the specific activity of cellular isoperoxidase was found in 5 mM $NiCl_2$-treated cell, while $NiCl_2$ had no effect on the secretion of peroxidase into the medium. Various concentrations of KCl did not change peroxidase secretion, but 5 mM $ZnCl_2$ reduced peroxidase secretion greatly. The major secretory isoperoxidases stimulated by $CaCl_2$, NaCl and cellulase were composed of cationic isoperoxidases C2 and C3, which were found to be localized in the cell wall of rice by examination of the enzyme in the protoplast. Furthermore, the secretion rates of secretory isoperoxidases were increased rapidly when cellulase was treated in the absence of the osmotic stabilizer of 0.4 M mannitol. These results suggest that the stimulations of secretory isoperoxidase levels seem to be due to the stimulation of secretion into the culture medium of rice.

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Production of Artificial Seeds by Alginate-encapsulation of Rice Somatic Embryos (벼의 수화겔 인공종자 생산)

  • 정원중;민성란;송남희;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.183-186
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    • 1994
  • Somatic embryos derived from cell suspension cultures of rice were singly alginate-encapsulated to be used as artificial seeds. When placed on half strength MS solid medium,73% of the encapsulated somatic embryos were capable of germination Encapsulation per se did not affect the germination frequency of embryos. When incubated by wrapping with moistured non-sterile filter paper, 60% of the encapsulated somatic embryos germinated. However encapsulated zygotic embryos without endosperm showed a high germination frequency regardless of the sterility of the incubation conditions. The results suggest that a greater susceptibility of somatic embryos to contaminants is attributed to lower germination frequency of encapsulated somatic embryos in non-sterile conditions.

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Secondary Products in Cell Suspension Culture of Salix koreensis (버드나무(Salix koreensis) 현탁배양(懸濁培養) 세포(細胞)의 대사산물(代謝産物))

  • Park, Young Goo;Shin, Dong Ill;Lee, Sang Goo
    • Journal of Korean Society of Forest Science
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    • v.78 no.2
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    • pp.198-208
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    • 1989
  • Cell suspension cultures for Salix koreensis was well established at the supplements of 2, 4-D with cytokinin particulary the combination of 1.0 mg/l 2, 4-D with 0.1 mg/l of zeatin. These combined rates of phytohormones are also effective to callus induction from S, koreensis leaf and its multiplication. Cultured media exhibited the great inhibitory effect on the germination of rice, barnyard grass and lettuce seeds, indicating the presence of biologically active substances in media. Several phenolic compounds such as pyrogallol, sinapic acid, cinnamic acid, tannic + gallic and p-chlorobenzoic acid were detected in the cell suspension culture. The inhibitory effect exhibited by cultured media may be partly attributed to these phenalic compounds.

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DNA Delivery into Embryogenic Cells of Zoysiagrass(Zoysia japonica Steud.) and Rice(Oryza sativa L.) by Electroporation (Electroporation을 이용한 잔디(Zoysia japonica Steud.) 및 벼(Oryza sativa L.) 배발생세포로의 DNA 도입)

  • 박건환;최준수;윤충호;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.309-314
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    • 1994
  • To develop simple and efficient transformation methods of monocotyledonous plane, electroporation-mediated delivery of DNA into intact embryogenic cell clumps was investigated in zoysiagrass and rice. Calli of zoysiagrass, induced from 3-week-old immature embryos, were suspension-cultured in MS basic medium supplemented with 1.0 mg/t 2.4-D and used for elechuporation. Calli, derived from immature inflorescences of 20 mm lenth of rice, were also suspension-cultured on N6 basic medium supplemented with 1.0 mg/L 2.4-D. Suspension-cultured embryogenic cell clumps were electroporated in liqid MS medium added with a Plasmid DNA (30 $\mu\textrm{m}$/ml), pGA1074, encoding ${\beta}$-glucuronidiase (GUS). DNA delivery into the cells through cell walls and cell membrane was confirmed by the transient expression of the GUS gene. Cell clumps of zoysiagrass and rice, electroporated with 400 volt at 800 pF capacitance, expressed GUS gene activity at a mean frequency of 25 units (one unit = one clony of blue cells) per 200 ${\mu}\ell$ of packed cell volume. Untreated cells and healed non-embryogenic cells did not exhibit GUS activity These results indicate that electroporation-mediated transformation can use intact embryogenic cells (thus avoiding the use protoplasts) in zoysiagrass and rice.

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