2007 Fiscal Year Final Research Report Summary
Functional genomics in the rice blastfungus, Magnapathe oryzae using an RNA siencing approach
| 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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| Keywords | RNAi / RNA silencing / Maonaoorthe orvzae |
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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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[Journal Article] (2007). Speciation in Pyricularia inferred from multilocus phylogenetic analysis
Author(s)
Hirata, K, Kusaba, M, Chuma, I, Osue, J, Nakayashiki, H, Mayama, S, Tosa, Y
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Journal Title
Mycol Res 111
Pages: 799-808
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] (2007). siRNAdependent and -independent posttranscriptional cosuppression of the LTR-retrotransposon MAGGY in the phytopathogenic fungus Magnaporthe oryzae
Author(s)
Murata, N, Kadotani, N, Yamaguchi, M, Tosa, Y, Mayama, S, Nakayashiki, H
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Journal Title
Nucleic Acids Res 35
Pages: 5987-5994
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] (2008)Systematic functional analysis of calcium signaling proteins in the genome of the rice blast fungus, Magnaporthe oryzae, using a high-throughput RNA silencing system
Author(s)
Nguyen, B, Q, Kadotani, N, Kasahara, S, Tosa, Y, Mayama, S, Nakayashiki, H
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Journal Title
Molecular Microbiology 68
Pages: 1348-1365
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] (2007)Analysis of calcium signaling proteins in the rice blast fungus, Magnaporthe oryzae, using an RNA silencing approach
Author(s)
Quoc, N, B, Kadotani, N, Tosa, Y, Mayama, S, Nakayashiki, H
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Journal Title
4th International Rice blast conference. Changsha, China. Abstract 41
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Analysis of calcium signaling proteins in the rice blast fungus, Magnaporthe oryzae, using an RNA silencing approach2007
Author(s)
Quoc, N.B., Kadotani, N., Tosa, Y., Mayama, S., Nakayashiki, H
Organizer
4th International Rice blast conference (Abstract P. 41)
Place of Presentation
Changsha, China
Year and Date
20070000
Description
「研究成果報告書概要(欧文)」より
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