| Project/Area Number |
01550763
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
発酵工学
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| Research Institution | Osaka University |
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Principal Investigator |
TAYA Masahito Osaka Univ., Dept. of Chem. Eng., Associate Professor, 基礎工学部, 助教授 (60144127)
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| Co-Investigator(Kenkyū-buntansha) |
TONE Setsuji Osaka Univ., Dept. of Chem. Eng., Professor, 基礎工学部, 教授 (40029418)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Plant Cell Culture / Plant Hairy Root / Bioreactor / Agrobacterium rhizogenesis / Metabolite Production / Peroxidase / Superoxide Dismutase / Pigment / 毛状根細胞 / アグロバクテリウムリゾゲネス / ス-パ-オキシドジスムタ-ゼ生産 / 色素生産 / バイオアクタ- / パ-オキシダ-ゼ生産 |
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| Research Abstract |
The induction of hairy roots was tried from various plants and their culture characteristics were investigated in biotechnological a spaects. The following results were obtained.
1. By infecting the speciments of carrot, horseradish, puk-bung and red beet with Agrobacteria rhizogenes, the hairy root clones were induced from these plants. Especially, puk-bung hairy root had high activity of peroxidase (valuable as diagnostic enzymes) and its activity was about 5 time that of original plant.
2. The growth rates of hairy roots were expressed in terms of branching frequency, elongation rate and decay rate of growing point (meristem) at the tips. It was possible to evaluate the culture properties of hairy roots based on this kinetic growth model. Moreover, the amounts of cell mass and main inorganic nutrients (K^+, NH<_4^+ and NO_3^- could be estimated during the hairy root cultures by the monitoring of medium conductivity.
3. The hairy roots from pak-bung and horseradish had high activities of Super-Oxide Dismutase (SOD). A two-step culture of pak-bung hairy root was performed on the basis of the nitrogen source effect on its growth. Namely, medium with NO_3^- was used in cell growth phase and the medium was replaced with NH_4^+ medium in SOD production phase. In this culture system, the SOD productivity was 5-7 times as high as that in the culture with medium containing only NO_3^- as a nitrogen source.
4. In the culture of red beet hairy root, significant amounts of intracellular pigments were secreted into medium by the interrupted limitation of oxygen supply. The repeated-batch culture of beet hairy root was carried out with the temporary oxygen starvation, and 20-25 % of the total pigments in cells was recovered from culture broth. Moreover, it was found that styrene-divinylbenzen resin was applicable to the pigment recovery from culture broth as an adsorbent.
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