| Project/Area Number |
09356002
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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 | 展開研究 |
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| Research Field |
植物保護
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
YAMADA Tetsuji Okayama University, Faculty of Agriculture, Professor, 農学部, 教授 (00191320)
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| Co-Investigator(Kenkyū-buntansha) |
ICHINOSE Yuki Okayama University, Graduate School of Natural Science and Technology, Associate Professor, 大学院自然科学研究科, 助教授 (50213004)
TAHARA Makoto Okayama University, Faculty of Agriculture, Professor, 農学部, 教授 (50274014)
OZAWA Hidenori Japan Tobacco Inc., Leaf Tobacco Research Laboratory, Researcher, 葉たばこ研究所, 研究員
田中 博 日本たばこ, 葉たばこ研究所, 主任研究員
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥17,600,000 (Direct Cost: ¥17,600,000)
Fiscal Year 1999: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1998: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1997: ¥9,700,000 (Direct Cost: ¥9,700,000)
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| Keywords | lysozyme / lytic enzyme / tobacco / phenylalanine ammonia-lyase / cis-element / フェニルアラニンアンモリアリアーゼ / 枯れ病菌 / リゾチーム遺伝子 / PAL / フェニールアラニンアンモニア・リアーゼ / エンドウ / トランスジェニック / ナス科植物 / 青枯れ病 / 遺伝子組換え |
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| Research Abstract |
Ralstonia solanacearum is a causal agent of bacterial wilt mainly to solanacearum plants. Although many approaches were made to control this pathogen, a sure method to protect the plants was not established yet. In this study, to breed genetically resistant solanacearum plants to this pathogen, we planed to generate the transgenic tobacco plants expressing the lysozyme gene isolated from the bacteriophage of this bacterium. Furthermore we utilized pea PSPAL1 promoter which shows root spec and pathogen-inducible expression pattern for the control of lysozyme gene.
We considered that the following results include the significant achievements in this research.
1. From bacteriophage P4282 of Ralstonia solanacearum, we isolated lysozyme protein and correspond gene, and structurally characterized.
2. We constructed the repeat promoter consisting the regulatory cis-elements (Box I, Box, II and Box IV of PSPAL1 promoter that expressed extremely high promoter activity. Especially the repeat promoter exhibited 6.5 hold activity than CaMV 35S promoter in roots.
3. Each CaMV 35S promoter, PSPAL1 promoter or the repeat promoter described above was fused with lysozyme gene, and transformed tobacco plants.
4. The transgenic tobacco plants that integrated chimeric lysozyme gene expressed higher lytic enzyme activity.
5. By the inoculation of 35S :: Lux gene cassette introduced R. solanacearum on the transgenic tobacco plants possessing PSPAL1 promoter :: GUS reporter gene enabled to establish the monitoring system to investigate the host defense response and behavior of bacterial pathogen simultaneously.
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