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Visualization of mitotic chromosomes in filamentous fungi by fluorescence staining and fluorescence in situ hybridization

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Abstract

Mitotic chromosomes of the plant pathogenic filamentous fungi Botrytis cinerea and Alternaria alternata were observed. Chromosomes prepared by the germ tube burst method were stained with the fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI) to yield figures with good resolution. Using this method, component chromosomes were clearly distinguished and the chromosome number could be determined. Fluorescence in situ hybridization (FISH) was also successfully applied to the specimens, revealing one ribosomal RNA gene cluster, or nucleolus organizer region (NOR) in the genome of each fungus. A long attenuated chromatid thread expanding from a condensed metaphase chromosome, which had been called a thread-like structure in B. cinerea, was proved to be an NOR. This is the first report of the successful application of FISH to the chromosomes of filamentous fungi.

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References

  • Francis DM, Michelmore RW (1993) Two classes of chromosome-sized molecules are present in Bremia lactucae. Exp Mycol 17:284–300

    Google Scholar 

  • Heath IB (1978) Experimental studies of mitosis in the fungi. In: Heath IB (ed) Nuclear division in the fungi. Academic Press, New York San Francisco London, pp 89–176

    Google Scholar 

  • Heath IB (1980) Variant mitoses in lower eukaryotes: Indicators of the evolution of mitosis? Int Rev Cytol 64:1–80

    Google Scholar 

  • Johnson GD, Araujo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350

    Google Scholar 

  • Kuroiwa T, Kojima H, Miyakawa I, Sando N (1984) Meiotic karyotype of the yeast Saccharomyces cerevisiae. Exp Cell Res 153:259–265

    Google Scholar 

  • Kuroiwa T, Miyamura S, Kawano S, Hizume M, Tho-e A, Miyakawa I, Sando N (1986) Cytological characterization of NOR in the bivalent of Saccharomyces cerevisiae. Exp Cell Res 165:199–206

    Google Scholar 

  • Li S, Harris CP, Leong SA (1993) Comparison of fluorescence in situ hybridization and primed in situ labeling methods for detection of single-copy genes in the fungus Ustilago maydis. Exp Mycol 17:301–308

    Google Scholar 

  • Maniatis T, Fntsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Mills D, McCluskey K (1990) Electrophoretic karyotypes of fungi: the new cytology. Mol Plant-Microbe Interact 3:351–357

    Google Scholar 

  • Murata M, Nakata N, Yasumuro Y (1992) Origin and molecular structure of a midget chromosome in a common wheat carrying rye cytoplasm. Chromosoma 102:27–31

    Google Scholar 

  • Perkins DD (1962) Preservation of Neurospora stock cultures with anhydrous silica gel. Can J Microbiol 8:591–594

    Google Scholar 

  • Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934–2938

    Google Scholar 

  • Raju NB (1986) A simple fluorescent staining method for meiotic chromosomes of Neurospora. Mycologia 78:901–906

    Google Scholar 

  • Scherthan H, Loidl J, Schuster T, Schweizer D (1992) Meiotic chromosome condensation and pairing in Saccharomyces cerevisiae studied by chromosome painting. Chromosoma 101:590–595

    Google Scholar 

  • Schwartz DC, Cantor CR (1984) Separation of yeast chromosome-sized DNAs by pulsed-field gradient gel electrophoresis. Cell 37:67–75

    Google Scholar 

  • Shirane N, Masuko M, Hayashi Y (1988) Nuclear behavior and division in germinating conidia of Botrytis cinerea. Phytopathology 78:1627–1630

    Google Scholar 

  • Shirane N, Masuko M, Hayashi Y (1989) Light microscopic observation on nuclei and mitotic chromosomes of Botrytis species. Phytopathology 79:728–730

    Google Scholar 

  • Skinner DZ, Budde AD, Leong SA (1991) Molecular karyotype analysis of fungi. In: Bennett JW, Lasure LL (ed) More gene manipulations in fungi. Academic Press, San Diego, pp 86–103

    Google Scholar 

  • Tsuge T, Kobayashi H, Nishimura S (1989) Organization of ribosomal RNA genes in Alternaria alternata Japanese pear pathotype, a host-selective AK-toxin-producing fungus. Curr Genet 16:267–272

    Google Scholar 

  • Umesono K, Hiraoka Y, Toda T, Yanagida M (1983) Visualization of chromosomes in mitotically arrested cells of the fission yeast Schizosaccharomyces pombe. Curr Genet 7:123–128

    Google Scholar 

  • Uzawa S, Yanagida M (1992) Visualization of centromeric and nucleolar DNA in fission yeast by fluorescence in situ hybridization. J Cell Sci 101:267–275

    Google Scholar 

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Taga, M., Murata, M. Visualization of mitotic chromosomes in filamentous fungi by fluorescence staining and fluorescence in situ hybridization. Chromosoma 103, 408–413 (1994). https://doi.org/10.1007/BF00362285

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  • DOI: https://doi.org/10.1007/BF00362285

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