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Molecular cloning of recalcitrant plant viruses genes by a modified RAPD method using RF-dsRNA

Research Project

Project/Area Number 12660041
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物保護
Research InstitutionUtsunomiya University

Principal Investigator

NATSUAKI Tomohide  Utsunomiya Univ., Fac.Agri., Professor, 農学部, 教授 (10134264)

Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordsrecalcitrant plant viruses / replicative form dsRNA / PCR / simple detection method / molecular cloning / 難純化RNAウイルス / キュウリ黄化ウイルス / tomato infectious chlorosis virus / Cryptovirus / カンキツトリステザウイルス
Research Abstract

Molecular cloning and sequencing of plant virus genes has been carried out during the past decade. One objective of molecular analysis of plant viruses has been the improvement of the methods to detect and diagnose pathogenic viruses. As templates for Cdna synthesis, nucleic acids are usually extracted from purified virus preparations in relatively pure form and in rather large amounts. Theses strategies rely on the purification of virus particles from infected plants. However, there are many recalcitrant viruses that can not be purified by current methods and, therefore, the standard nucleic acid templates are not accessible for their cloning. It is the viruses for which there are no available antisera that alternate methods of detection and diagnosis are needed. For several of these viruses, the application of dsRNA extraction techniques from herbaceous or woody host plants has permitted the detection of viral replicative nucleic acids (RF-dsRNA). The main objective of this research was the production of cDNA clones generated from RF-dsRNA extracted from virus-infected plant tissues. In this research the simple one tube RT-PCR method to detect these viruses was established. Furthermore, full nucleotide sequences of mild and severe isolates of citrus tristeza vireus and Cucumber yellows virus were determined. Tomato infectious chlorosis virus, a member of Criniviruses, was newly recognized in Japan.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Suastika, G., Natsuaki, T., et al.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67. 73-77 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hartono, S., Natsuaki, T., et al.: "Yellowing disease of tomatoes caused by Tomato infectious chlorosis virus newly recognized in Japan"Journal of General Plant Pathology. 69. 61-64 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hartono, S., Natsuaki, T., et al.: "Nucleotide sequence and genome organization of cucumber yellows virus, a member of the genus Crinivirus"Journal of General Virology. 84. 1007-1012 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Suastika,G., Natsuaki,T., Terui,H., Kano,T., Ieki,H. and Okuda,S.: "Nucleotide sequence of citrus tristeza virus seedlding yellows isolate"Journal of General Plant Pathology. 67-1. 73-77 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hartono, S., Natsuaki, T, Sayama, H., Atarashi, H. and Okuda, S.: "Yellowing disease of tomatoes caused by Tomato infectious chlorosis virus newly recognized in Japan"Journal of General Plant Pathology. 69-1. 61-64 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hartono, S., Natsuaki, T., Genda, Y. and Okuda, S.: "Nucleotide sequence and genome organization of cucumber yellows vinos, a member of the genus Crinivirus"Journal of General Virology. 84-4. 1007-1012 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Suastika, G., Natsuaki, T., Terui, H., Kano, T., Ieki, H., Okuda, S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67・1. 73-77 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hartono, S., Natsuaki, T., Sayama, H., Atarashi, H., Okuda, S.: "Yellowing disease of tomatoes caused by Tomato infectious chlorosis virus newly recognized in Japan"Journal of General Plant Pathology. 69・1. 61-64 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hartono, S., Natsuaki, T., Genda, Y., Okuda, S.: "Nucleotide sequence and genome organization of cucumber yellows virus, a member of the genus Crinivirus"Journal of General Plant Pathology. 84・4. 1007-1012 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Suastika, G., Natsuaki, T., Terui, H., Kano, T., Ieki, H., Okuda, S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67(1). 73-77 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Suastika,G.,Natsuaki,T.,Terui,H.,Kano,T.,Ieki,H.and Okuda,S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67(1). 73-77 (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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