| Project/Area Number |
15380002
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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 |
Breeding science
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| Research Institution | University of Tsukuba |
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Principal Investigator |
EZURA Hiroshi University of Tsukuba, Graduate School of Life and Environmental Sciences, Professor, 大学院生命環境科学研究科, 教授 (00332552)
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| Co-Investigator(Kenkyū-buntansha) |
MINAMISAWA Kiwamu Tohoku University, Graduate School of Life Sciences, Professor, 大学院生命科学研究科, 教授 (70167667)
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| Project Period (FY) |
2003 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2006: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2005: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2004: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2003: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | Cucurbitacae / gene transfer / Agrobacterium / transgenic plant / ethylene / biosynthesis / rizobitoxin / ACC deaminase / ACCダアミナーゼ / 組替え体 |
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| Research Abstract |
The objectives of the research were 1) to elucidate the plant-Agrobacterium interaction through ethylene signaling pathway and 2) to apply the knowledge on molecular breeding of Agrobacterium which shows higher ability of gene transfer to Cucurbitacae plants. The followings are the major results obtained.
1. We observed that ethylene itself did not inhibit Agrobacterium cell growth whereas plants responding to ethylene, that inhibiting the vir gene expression in Agrobacterium on the plant cells. It is likely that the inhibitory effect is caused by materials which are induced/depressed by ethylene in the plant cells. These results allow us to speculate that we can develop super-Agrobacterium which shows higher ability of gene transfer by providing the ability to inhibit ethylene biosynthesis in plant cells.
2. Rhizobitoxine is an ethylene biosynthesis inhibitor produced by soybean root nodule bacteria (Bradyrhizobium elkanii), and reduces ethylene production from the host plants. rtxACDEFG genes from B. elkanii and OAH synthase gene from Mesorhizobium loti were introduced to Agrobacterium tumefaciens C58 strain, and the genetically engineered Agrobaterium produced rhizobitoxine. ACC deaminase is an enzyme produced by symbiotic bacteria, and degrade ACC which results in inhibition of ethylene biosynthesis in plants. ACC deaminase gene from Pseudomonas strain was introduced to A. tumefaciens C58 strain, and the genetically engineered Agrobacterium showed ACC deaminase activity.
3. The genetically modified Agrobacterium producing either rhizobitoxineor ACC deaminase showed higher ability of gene transfer to melon. In addition, the Agrobacterium producing ACC deaminase also showed higher ability of gene transfer to Arabidopsis.
In conclusion, we are succeeded in molecular breeding to production of so-called super-Agrobacterium which shows higher ability of gene transfer into plant cells without inhibition of vir gene expression.
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