| Project/Area Number |
63480045
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物保護
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| Research Institution | Kobe University (1990)
Kagawa University (1988-1989) |
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Principal Investigator |
MAYAMA Shigeyuki Kobe University, Dept. of Plant Protection, Assistant Professor, 農学部, 助教授 (00112251)
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| Co-Investigator(Kenkyū-buntansha) |
TOYODA Hideyioshi Kinki University, Dept. of Agronomy, Assistant Professor, 農学部, 助教授 (00150805)
京 正晴 香川大学, 農学部, 助手 (70195395)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1989: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Agrobacterium rhizogenes / transformed plant / melon / tomato / hairy root / genetic engineering / プロトプラスト / エレクトロポ-レ-ション / 形質転換 / Riプラスミド / 病害抵抗性 / 植物病原毒素 / 毒素感受性遺伝子 / 抵抗性遺伝子解析 / Tiプラスミドベクター |
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| Research Abstract |
Utilization of hairy root induced by T-DNA insertion with Agrobacterium rhizogenes was studied to elucidate resistance or susceptibility mechanisms in plant disease and to induce resistance by gene-transfer.
Hairy roots of melon were induced by A. Rhizogenes-infection. The hairy roots could grow vigorously on hormone-free medium. Hairy roots were differentiated into callus tissues when cultured in the presence of growth regulator, and shoots were frequently redifferentiated from the callus when the callus tissues were subcultured in the 50 ug/l of 2, 4-D and BAP. pBI121 was introduced into leaf cell by A. Rhizogenes, and both NPT II and GUS gene were expressed in developed hairy roots. Southern blot analysis of DNA from GUS-positive hairy root lines confirmed the integration of vector DNA to the plant chromosome. Callus tissues, leaves developed from shoots, and secondary hairy roots of regenerants were treated with kanamycin, and then subjected to histochemical detection of b-guluculonidase activity. The results clearly indicated that both the NPT vector and GUS genes transferred to the primry hairy roots were stably integrated and expressed at all stages of redifferentiation. Coat protein gene of Cucumber mosaic virus was introduced in melon cells through agroinfection, and the integration of CP gene was demonstrated by Southern blot analysis of DNA from transformed callus.
Hairy roots of tomato were also induced by A. Rhizogenes. Transformed rate of the roots by T-DNA were over 50% of the induced. Screening of hairy roots resistant to Fusarium oxysporum lycopercisi was not successful.
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