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Studies on pathogenicity and soil-transmissibility of plant RNA viruses in the genus Furovirus

Research Project

Project/Area Number 16380034
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Plant pathology
Research InstitutionThe University of Tokyo

Principal Investigator

SHIRAKO Yukio  The University of Tokyo, Asian Natural Environmental Science Center, Professor, アジア生物資源環境研究センター, 教授 (90143023)

Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2006: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2005: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2004: ¥6,400,000 (Direct Cost: ¥6,400,000)
KeywordsGenus Furovirus / Positive-sense RNA virus / Soil-borne wheat mosaic virus / Pathogenicity / Temperature sensitivity / Soil transmissibility / Yeast two-hybrid system / Chromosomal integration / 欠失変異 / Furovirus / 土壌伝染 / Polymyxa graminis / プロトプラスト
Research Abstract

1. Soil-borne wheat mosaic virus (SBWMV), the type species of the Genus Furovirus, forms spontaneous deletion mutations in RNA2 in infected wheat plants. Detailed analysis of deleted sites in RNA2 showed that the 3' termini of deletions end immediately upstream of the p19 promoter sequence. Deletion further into the promoter region inhibited transcription of p19 subgenomic RNA and cell-to-cell movement of viral RNA although RNA replication in single cells was not affected.
2. The p37 gene on RNA1 and CP, RT and p19 genes on RNA2 were replaced with a green fluorescence protein (GFP) gene in infectious cDNA clones of SBWMV. In vitro transcripts were inoculated to wheat leaves and GFP fluorescence in inoculated leaves and upper leaves were examined with a fluorescence light microscopy. Results showed that p37 is the viral RNA movement protein, which p19 is required for efficient cell-to-cell movement and long-distance movement of viral RNA, and that CP is required for persistent infection of the virus. CP-RT was shown to facilitate movement of viral RNA systemically.
3. The wild-type SBWMV has optimal temperatures for infection below 20℃. However, when systemically infected wheat or barley plants were kept at 25℃, virus variants that infects plants systemically at 25℃ appeared in limited plants. When the variant genome was thoroughly analyzed, only one asynonymous nucleotide change was found in p37 movement protein gene. The mutation in p37 was confirmed to be responsible for persistent infection of SBWMV variants at 25℃.
4. Increased virulence caused by mutants of SBWMV was mapped and identified in p19 gene on RNA2.
5. A conventional yeast two-hybrid system was modified by integration of BD:bait genes into yeast chromosome. Efficiency to get true-positive clones from cDNA library screening was significantly increased.

Report

(4 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report

Research Products

(8 results)

All 2006 2005

All Journal Article (7 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Orchid fleck virus is a rhabdovirus with an unusual bipartite genome.2006

    • Author(s)
      Kondo, H., Maeda, T., Shirako, Y., Tamada, T.
    • Journal Title

      Journal of General Virology 87

      Pages: 2413-2421

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] RNase E maintenance of proper FtsZ/FtsA ratio required for nonfilamentous growth of Escherichia coli cells but not for colony-forming ability.2006

    • Author(s)
      Tamura, M., Lee, K., Miller, C.A., Moore, C.J., Shirako, Y., Kobayashi, M., Cohen, S.N.
    • Journal Title

      Journal of Bacteriology 188

      Pages: 5145-5152

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Orchid fleck virus is a rhabdovirus with an unusual bipartite genome2006

    • Author(s)
      Kondo, H., Maeda, T., Shirako, Y., Tamada, T.
    • Journal Title

      Journal of General Virology 87

      Pages: 2413-2421

    • Related Report
      2006 Annual Research Report
  • [Journal Article] RNase E maintenance of proper FtsZ/FtsA ratio required for nonfilamentous growth of Escherichia coli cells but not for colony-forming ability2006

    • Author(s)
      Tamura, M., Lee, K., Miller, C.A., Moore, C.J., Shirako, Y., Kobayashi, M., Cohen, S.N.
    • Journal Title

      Journal of Bacteriology 188

      Pages: 5145-5152

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Stable deletions arising in the readthrough region of Soil-borne wheat mosaic virus RNA2 define the 5' limit of the functional promoter for the p19 subgenomic RNA.2005

    • Author(s)
      Yamamiya, A., Miyashini, M., Shirako, Y.
    • Journal Title

      Archives of Virology 150

      Pages: 1871-1884

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Stable deletions arising in the readthrough region of Soil-borne wheat mosaic virus RNA2 define the 5' limit of the functional promoter RNA for the p19 subg2005

    • Author(s)
      Yamamiya, Y., Miyanishi, M., Shirako, Y
    • Journal Title

      Archives of Virology 150

      Pages: 1871-1884

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Stable deletions arising in the readthrough region of Soil-borne wheat mosaic virus RNA2 define the 5' limit of the functional promoter RNA for the p19 subgenomic2005

    • Author(s)
      Yamamiya, Y., Miyanishi, M., Shirako, Y
    • Journal Title

      Archives of Virology 150(印刷中)

    • Related Report
      2004 Annual Research Report
  • [Patent(Industrial Property Rights)] タンパク質結合検出方法2006

    • Inventor(s)
      白子幸男, 宮下脩平
    • Industrial Property Rights Holder
      国立大学法人 東京大学
    • Industrial Property Number
      2006-281333
    • Filing Date
      2006-10-16
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary

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Published: 2004-03-31   Modified: 2016-04-21  

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