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1996 Fiscal Year Final Research Report Summary

Development of the cytogenetic techniques to analyze chromosome rearrangements in plant-pathogenic filamentous fungi

Research Project

Project/Area Number 07660059
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

TAGA Masatoki  Okayama University, Facullty of Science, Associate Professor, 理学部, 助教授 (80236372)

Co-Investigator(Kenkyū-buntansha) MURATA Minoru  Okayama University, Research Institute for Bioresources, Associate Professor, 資源生物科学研究所, 助教授 (20166292)
Project Period (FY) 1995 – 1996
Keywordsplant-pathogenic fungi / filamentous fungi / chromosome / fluorescence in situ hybridization / pulsed field gel electrophoresis / cytogenetics
Research Abstract

1.Background and objectives
One of the major causes for genetic variation of plat-pathogenic filamentous fungi is the change of complement chromosomes of the genomes. Such chromosomal changes are thought to be constituted of aneuploidy and structural rearrangements of chromosomes such as translocation and inversion. Presently techniques that enable detailed cytogenetic analyzes are not available for filamentous fungi. The purpose of this study is to develope novel cytogenetic techniques to elucidate chromosomal changes, especially the rearrangements, in plant fungal pathogens. We set up three concrete goals, that is, (1) application of fluorescence in situ hybridization (FISH) to identify specific chromosomes, (2) detection of chromosomal rearrangements by FISH and pulsed field gel electrophoresis (PFGE), (3) search for the kinds and frequencies of chromosomal rearrangements in fungal populations.
2.Materials
Three plant pathogens, Fusarium solani f.sp.pisi, Botrytis cinerea and Alternari … More a alternata, were used.
3.Results and conclusions
(1)FISH and PFGE techniques
Experimental protocols for FISH and PFGE were successfully developed especially for F.solani. Using our PFGE prosocol the chromosomes in size range between ca.6-Mb and ca.500-kb could be clearly separated in a sigle run, which enabled us to analyze karyotypic polymorphisms of F.solani. Regarding FISH four types of probes, that is, plasmid cosmid, total nuclear DNA,PCR-amplified DNA of specific chromosomes were labeled with either biotin or DIG.With these probes specific chromosomes became detectable among the somatic metaphase complement chromosomes of the genome.
(2)Identification and characterization of specific chromosomes
Nucleolar-organizing chromosomes was identified by FISH and PFGE using rDNA as a probe in F.solani, A.alternata and B.cinerea. The number of nucelolar-organizing chromosomes was only one in F.solani and B.cinerea, while two were detected in A.alternata. The morphological change of rDNA in the course of nuclear division was evident for F.solani.
B chromosome in F.solani was detected by three types of FISH (ordinary FISH with B chromosome-specific cosmid clone, genomic in situ hybridization, and chromosome painting). This was the first success of cytological detection and visualization of B chromosome in fungi.
Simultaneous identification of two chromosomes was accomplished by two-color FISH using probes of plasmid-cosmid clones or chromosome painting coupled with hybridization detection by FITC and rhodamine.
In this study we have not succeeded in showing the chromosomal rearrangement yet mainly because of the shortage of time. Less

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] M.Taga et al: "Comparison of different karyotyping methods in filamenfous ascomycetes-a case study of Nectria haemafococca (anamorph:Fusarium solani)" Mycological Research. (印刷中). (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 多賀 正節 ら: "蛍光 in situ ハイブリダイゼーションによる Nectria haematococca 体細胞染色体の分子細胞遺伝学的解析" 日本植物病理学会報. 62. 299 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 赤松 創 ら: "パルスフィールドゲル電気泳動法(PFGE)および蛍光染色法によるAlternaria属、植物病原菌の染色体解析" 日本植物病理学会報. 63. 250-251 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 多賀 正節 ら: "Nectria haematococca 体細胞染色体の染色体ペインティング" 日本植物病理学会報. 63. 251 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Taga, M.et al.: "Comparison of different karyotyping methods in filamentous ascomycetes -a case study of Nectria haematococca (anamorph : Fusarium solani)." Mycological Research. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taga, M.et al.: "Molecular cytogenetical analysis of somatic chromosomes of Nectria haematococca by fluorescence in situ hybridization. (in Japanese)" Ann.Phytopathol.Soc.Jpn.62. 299 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Taga, M.et al.: "Chromosome painting of Nectria haematococca somatic chromosomes. (in Japanese)" Ann.Phytopathol.Soc.Jpn.63. 251 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akamatsu, H.et al.: "Chromosome analyzes of Alternaria fungi by pulsed field gel electrophoresis and fluorescence staining. (in Japanese)" Ann.Phytopathol.Soc.Jpn.63. 250-251 (1997)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-16  

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