| Project/Area Number |
17380030
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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 |
Plant pathology
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| Research Institution | Kobe University |
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Principal Investigator |
NAKAYASHIKI Hitoshi Kobe University, Graduate School of Agricultural Science, Associate Professor (50252804)
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| Co-Investigator(Kenkyū-buntansha) |
MAYAMA Shigeyuki Kobe University, Graduate School of Agricultural Science, Professor (00112251)
TOSA Yukio Kobe University, Graduate Schcol of Agricultural Science, Professor (20172158)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,870,000 (Direct Cost: ¥15,700,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2007: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥8,300,000 (Direct Cost: ¥8,300,000)
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| Keywords | RNAi / RNA silencing / Maonaoorthe orvzae / ポストゲノム / 病原性遺伝子 |
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| Research Abstract |
We developed an RNA silencing vector, pSilent-Duall (pSD1), with a convergent dual Pol II promoter system that provides a high-throughput platform for functional genomics research in filamentous fungi. In the pSD1 system, the target gene was designed to be transcribed as a chimeric RNA with enhanced green fluorescent protein (eGFP) RNA. This enabled us to efficiently screen the resulting transformants using GFP fluorescence as an indicator of gene silencing. A model study with the eGFP gene showed that pSD1-based vectors induced gene silencing via the RNAi pathway with slightly lower efficiency than did hairpin eGFP RNA-expressing vectors. To demonstrate the applicability of the pSD1 system for elucidating gene function in the rice blast fungus Magnaporthe oryzae, 37 calcium signaling-related genes that include almost all known calcium signaling proteins in the genome were targeted for gene silencing by the vector. Phenotypic analyses of the silenced transformants showed that at least 26, 35, and 15 of the 37 genes examined were involved in hyphal growth, sporulation and pathogenicity, respectively, in M oryzae. These included several novel findings such as that Pmcl-, Spfl- and Neo1-like Ca^<+>2 pumps, calreticulin, and calpactin heavy chain were essential for fungal pathogenicity.
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