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Functional genomics in the rice blastfungus, Magnapathe oryzae using an RNA siencing approach

Research Project

Project/Area Number 17380030
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Plant pathology
Research InstitutionKobe University

Principal Investigator

NAKAYASHIKI Hitoshi  Kobe University, Graduate School of Agricultural Science, Associate Professor (50252804)

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)
Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
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)
KeywordsRNAi / RNA silencing / Maonaoorthe orvzae / ポストゲノム / 病原性遺伝子
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.

Report

(4 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (22 results)

All 2008 2007 2006 2005 Other

All Journal Article (17 results) (of which Peer Reviewed: 6 results) Presentation (2 results) Book (3 results)

  • [Journal Article] Systematic functional analysis of calcium signaling proteins in the genome of the rice blast fungus, Magnaporthe oryzae, using a high-throughput RNA silencing system.2008

    • Author(s)
      Nguyen, et. al.
    • Journal Title

      Molecular Microbiology 68

      Pages: 1348-1365

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] RNA interference: roles in fungal biology.2008

    • Author(s)
      Nakayashiki, et. al.
    • Journal Title

      Current Opinion in Microbiology (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Speciation in Pyricularia inferred from multilocus phylogenetic analysis.2008

    • Author(s)
      Hirata, et. al.
    • Journal Title

      Mycol Res. 111

      Pages: 799-808

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Systematic functional analysis of calcium signaling proteins in the genome of the rice blast fungus, Magnaporthe oryzae, using a high-throughput RNA silencing system.2008

    • Author(s)
      Nguyen, et. al.
    • Journal Title

      Molecular Microbiology (In press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] siRNA-dependent and-independent posttranscriptional cosuppression of the LTR-retrotransposon MAGGY in the phytopathogenic fungus Magnaporthe oryaze.2007

    • Author(s)
      Murata, et. al.
    • Journal Title

      Nucleic Acids Res. 35

      Pages: 5987-5994

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] siRNA-dependent and -independent posttranscriptional cosuppression of the LTR-retrotransposon MAGGY in the phytopathogenic fungus Magnaporthe oryzae.2007

    • Author(s)
      Murata, et. al.
    • Journal Title

      Nucleic Acids Res. 35

      Pages: 5987-5994

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Evolution and diversification of RNA silencing proteins in fungi2006

    • Author(s)
      Nakayashiki H. et al.
    • Journal Title

      J. Mol. Evol 63

      Pages: 127-135

    • NAID

      120000949568

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Specific cleavage of ribosomal RNA and mRNA during victorin-induced apoptotic cell death in oat2006

    • Author(s)
      Hoat, T. et al.
    • Journal Title

      Plant J. 46

      Pages: 922-933

    • Related Report
      2006 Annual Research Report
  • [Journal Article] The C-terminal chromodomain-like module in the integrase domain is crucial for high transposition efficiency of the retrotransposon MAGGY2005

    • Author(s)
      Nakayashiki, H. et al.
    • Journal Title

      FEBS letters 579

      Pages: 488-492

    • NAID

      120000942509

    • Related Report
      2005 Annual Research Report
  • [Journal Article] RNA silencing as a tool for exploring gene function in Ascomycete fungi2005

    • Author(s)
      Nakayashiki, H. et al.
    • Journal Title

      Fungal Genet. & Biol. 42

      Pages: 275-283

    • NAID

      120000942198

    • Related Report
      2005 Annual Research Report
  • [Journal Article] RNA silencing in fungi : Mechanisms and applications2005

    • Author(s)
      Nakayashiki H
    • Journal Title

      FEBS letters 579

      Pages: 5950-5957

    • NAID

      120000943890

    • Related Report
      2005 Annual Research Report
  • [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
    • Journal Title

      Mycol Res 111

      Pages: 799-808

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [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
    • Journal Title

      Nucleic Acids Res 35

      Pages: 5987-5994

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [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
    • Journal Title

      Molecular Microbiology 68

      Pages: 1348-1365

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [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
    • 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
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Evolution and diversification of RNA silencing proteins in fungi

    • Author(s)
      Nakayashiki H. et al.
    • Journal Title

      J.Mol.Evol (In press)

    • NAID

      120000949568

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Specific cleavage of ribosomal RNA and mRNA during victorin-induced apoptotic cell death in oat

    • Author(s)
      Hoat, T. et al.
    • Journal Title

      Plant J. (In press)

    • Related Report
      2005 Annual Research Report
  • [Presentation] Analysis of calcium signaling proteins in the rice blast fungus, Magnaporthe oryzae, using an RNA silencing approach.2007

    • Author(s)
      Nguyen, et. al.
    • Organizer
      4th International Rice blast conference
    • Place of Presentation
      Changsha, China
    • Year and Date
      2007-10-11
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [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
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] RNAi in fungi. In: RNAi (Advanced Methods)2008

    • Author(s)
      Nakayashiki, H.
    • Total Pages
      138
    • Publisher
      Taylor and Francis. UK
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] RNAi in fungi. In: RNAi(Advanced Methods)2008

    • Author(s)
      Nakayashiki, H.
    • Total Pages
      138
    • Publisher
      Taylor and Francis. UK
    • Related Report
      2007 Annual Research Report
  • [Book] RNA interference (RNAi) (eds. M. Latterich)2007

    • Author(s)
      Nakayashiki, H.
    • Publisher
      Taylor and Francis. UK(in press)
    • Related Report
      2006 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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