| Project/Area Number |
11556027
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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 | 展開研究 |
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| Research Field |
林学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
HOGETSU Taizo Asian Natural Environmental Science Center, The University of Tokyo, Professor, アジア生物資源環境研究センター, 教授 (10107170)
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Katsumi Asian Natural Environmental Science Center, The University of Tokyo, Associate Professor, アジア生物資源環境研究センター, 助教授 (80211895)
IDE Yuji Graduate School of Agricultural and Life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (90213024)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,100,000 (Direct Cost: ¥13,100,000)
Fiscal Year 2001: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2000: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1999: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Cryptomeria japonica / Chamaecyparis obtusa / Pinus densiflora / PCR breeding marker / microsatellite (SSR) marker / surperssion PCR / マイクロサテライト(SSR) / gene walking 法 / 多型解析 / マイクロサテライト多型マーカー / ISSR / gene walking法 / メンブランフィルター / 共優性 / 種特異性 / ヘテロ接合率 |
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| Research Abstract |
The aim of this project was to develop microsatellite markers for breeding of the main silvicultural tree species, Pinus densiflora, Chamaecyparis obtusa and Cryptomeria japonica, and to establish a novel method for microsatellite development, which needs less time and labor. First of all we developed several microsatellite markers of P. densiflora and C. obtusa by an improved method including an enrichment process of microsatellite containing DNA fragments. On the other hand, we investigated a new technical idea, and established a novel simple method for development of microsatellite markers. This method was composed of two stages; in the first stage, a sequence neighboring a microsatellite was determined and -one primer was designed from the sequence. The second stage, a sequence including both sequences neighboring the microsatellite were determined by use of a suppression PCR technique, and designed the other primer for PCR amplification of the microsatellite region. In the first stage, a lot of sub-clones including different microsatellite regions could be obtained, indicating that this method is effective for developing a number of markers. This methods were also applicable to species of fungus insect, nematode in addition to the tree species. This method will enable mass production of microsatellite breeding markers for the main silvicultural tree species more easily.
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