| Project/Area Number |
07454222
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物生理
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| Research Institution | Japan Women's University |
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Principal Investigator |
KOMAMINE Atsushi Japan Wamen's University, Faculty of Science, Professor, 理学部, 教授 (90011494)
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| Co-Investigator(Kenkyū-buntansha) |
OGAWA Kyoko Japan Wamen's University, Faculty of Science, Assistant, 理学部, 助手 (60130738)
MAKI Hisae Japan Wamen's University, Faculty of Science, Assistant, 理学部, 助手 (70212206)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | somatic embryogenesis / gene expression / antisense DNA / tarticle gun / gene introduction / in situ hybridigation / carrot / 硝酸還元酵素 / ホウレンソウ |
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| Research Abstract |
1. Cloning and analysis of gene expressing in early stages of somatic embryogenesis in carrot suspension sultures
A gene, which was expressed specifically only in early stages of somatic embryogenesis, was isolated by substractive differential screening in carrot suspension cultures, it was cloned and designated CEM6.Sequence of nucleotides of CEM6 cDNA revealed that CEM6 encoded glycine-rich hydrophobic protein. Northern analysis indicated that CEM6 was expressed globular-torpedo shaped embryos and the expression was declined thereafter. However, in situ hybridization revealed that CEM6 was expressed only in peripheral cells and the expression was most clear in torpedo shaped embryos. It was interesting that CEM6 expression was disappeared immediately after treatment with 2,4-D of torpedo shaped embryos for 24hr.
2. Induction of somatic embryogenesis from single cells
Single cells were released from hypocotyls of regenerated plantlets in vitro. When these single cells were cultured in conditioned medium, they differentiated directly to embryos at high frequency. Thus, high frequency embryogenesis from single cells was established.
3. Analysis of the function of CEM6 by introduction of anti-sense CEM6
Introduction of anti-sense CEM6 fused with GUS and npt II genes was attempted in order to know the function of CEM6 by observing embryogenesis of transformed cells. Introduction of CEM6 was performed by bombardment using a particle gun. Stable transformant cells were obtained. Observation of embryogenesis of transformant cells is in progress.
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