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Molecular cytogenetic analysis of the genome of the potato late blight fungus Phytophthora infestans

Research Project

Project/Area Number 18580043
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

TAGA Masatoki  Okayama University, Graduate School of Natural Science and Technology, Associate Professor (80236372)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
Keywordspotato late blight / Phytophthora / chromosome / genome / Oomycete / molecular cytogenetics / ploidy / ジャガイモ疫病菌 / ジャガイモ / 疫病菌 / 細胞遺伝子
Research Abstract

Phytophthora infestans is the causal pathogen of the most devastating potato disease called late blight. Although genome sequence project of this fungus is under way, basic genome information from cytogenetics is very scarce. In this study, we conducted molecular cytogenetic analyses to determine the karyotype and ploidy of P. infestans.
1. Karyotype analysis
We developed a method to prepare mitotic chromosome specimens by combining the germ tube burst method and chemical treatment with colchicine, colcemid, and cyclohexamide. Chromosomes prepared with this method were stained with DAPI, Giemsa, and silver nitrate for karyotyping. Chromosome number of a standard diploid strain was 2n=10-14 in most specimens including one or two NOR chromosomes. Sister chromatids in a coiled shape and heterochromatines could be observed in not-fully condensed chromosomes.
2. Fluorescence in situ hybridization (FISH)
Probes were cloned and FISH was performed to identify specific chromosomes and analyze ploidy cytologically. With rDNA as a probe, NOR was located near the end of chromosome. Using single copy genes, NiaA and Piexol, as probes, the number of signals per nucleus was shown to indicate ploidy level.
3. Flow cytometry (FCM)
The method to measure nuclear DNA contents of P. infestans by FCM was established. Using Arabidopsis thaliana as the internal reference, genome size of P. infestans was estimated to be 250-300Mb. The survey of world-wide collection of ca. 60 strains showed that strains can be grouped into haploids, dipoids, triploids, aneuploids, and heterkaryons. Formerly reported tetraploids could not be detected in our collection.

Report

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

    (7 results)

All 2008 2007 2006

All Presentation (7 results)

  • [Presentation] 日本産ジャガイモ疫病菌の倍数性解析2008

    • Author(s)
      有本英里香, 金子真美, 多賀正節
    • Organizer
      平成20年度日本植物病理学会大会
    • Place of Presentation
      島根県松江市
    • Year and Date
      2008-04-26
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ploidy analysis of Japanese isolates of Phvtovhthora infestans.2008

    • Author(s)
      Arimoto, E., Taga, M.
    • Organizer
      Annual Meeting of the Phytopathological Society of Japan
    • Place of Presentation
      Matsue, Shimane, Japan
    • Year and Date
      2008-04-26
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Analysis of the genomic ploidy of Phytophthora infestans by flow cytometery and fluorescence in situ hybridization.2007

    • Author(s)
      Kaneko, M., Arimoto, E., Taga, M.
    • Organizer
      XXIV Fungal Genetics Conference
    • Place of Presentation
      Asilomar Conference Center, Pacific Grove, California, USA
    • Year and Date
      2007-03-23
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Analysis of the genomic ploidy of Phytophthora infestans by flow cytometery and fluorescence in situ hybridization.2007

    • Author(s)
      Kaneko, M., Arimoto, E., Taga, M.
    • Organizer
      XXIV Fungal Genetics Conference
    • Place of Presentation
      Asilomar Conference Center, Pacific Grove, CA, USA
    • Year and Date
      2007-03-23
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Analysis of the genomic ploidy of Phytophthora infestans by flow cytometery and fluorescence in situ hybridization2007

    • Author(s)
      Kaneko, M., Arimoto, E. and Taga, M.
    • Organizer
      XXIV Fungal Genetics Conference
    • Place of Presentation
      Asilomar Conference Center, Pacific Grove, California, USA
    • Year and Date
      2007-03-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] フローサイトメトリーによるジャガイモ疫病菌の核ゲノムサイズ解析2006

    • Author(s)
      金子真美, 多賀正節
    • Organizer
      平成18年度日本植物病理学会大会
    • Place of Presentation
      北海道札幌市
    • Year and Date
      2006-06-05
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Flow cytometric analysis of the genome size in Phytophthora infestans.2006

    • Author(s)
      Kaneko, M., Taga, M.
    • Organizer
      Annual Meeting of the Phytopathological Society of Japan
    • Place of Presentation
      Sapporo, Japan
    • Year and Date
      2006-06-05
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2006-04-01   Modified: 2016-04-21  

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