1996 Fiscal Year Final Research Report Summary
Mass production of regenerated plants by artificial seed system
| Project/Area Number |
06403019
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Nagoya University |
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Principal Investigator |
KOBAYASHI Takeshi School of Engineering Nagoya Vniversity Professor, 工学部, 教授 (10043324)
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| Co-Investigator(Kenkyū-buntansha) |
UOZUMI Nobuyuki Bioscience Center, Nagoya Vniversity Associate Professor, 生物分子応答研究センター, 助教授 (40223515)
HONDA Hiroyuki School of Engineering Nagoya Vniversity Associate Professor, 工学部, 助教授 (70209328)
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| Project Period (FY) |
1994 – 1996
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| Keywords | artificial seed / regeneration / hairy root / adventive embryo / mechanical cut / secondary metabolite / preservation / cultivation engineering |
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| Research Abstract |
Efficient prodution system of adventive embryo, automatic selection system of adventive embryo and artificial seed system from hairy roots were studied extensively.
Celery embryos and plantlets were found to be selectively released in a culture of immobilized Ca-alginate gel beads in which celery callus was entrapped under regeneration conditions. Repeated batch culture with 5 mM CaCl_2 provided long-term(more than 154d) embryo and plantlet production without gel beads disruption. Productivity of plantlets in the immobilized cell culture was 2,2-fold as high as that in the suspension culture.
An efficient plantlet production from horseradish hairy roots was studied to apply the hairy roots for artificial seed. To make the hairy root fragments, a blender was applied after the root growth. A fragmentation time of 30 s gave the best result on the plantlet formation. Treatment of the hairy roots with naphthaleneacetic acid(1.0 mg/l) made the culture time shorter and increased plantlet productivity. Treatment with kinetin at 0.1 mg/l after fragmentation yielded the highest number of plantlets.
Ajuga hairy root in which the beta-glucuronidase(GUS)gene was introduced under control of the tomato ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS3B) promoter was constructed by the Agrobacterium rhizogenes-mediated cotansformation. The GUS-transformed hairy root could also be efficiently regenerated into plantlets through this procedure. GUS activity was detected in leaf tissue of the regenerated plants.
For plant regeneration from fragmented tips of horseradish hairy root, a shaking-vessel type bioreactor was used. Number of regenerated plantlet depended on a rotational spped, and the maximum number was obtained at 120 rpm. It was concluded that the shaking-vessel type bioreactor was suitable for mass-production of regenerated plantlet.
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