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Meiotic transmission of a hypomethylated repetitive DNA family in tobacco

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Abstract

We have recently shown that hypomethylation of cytosine residues in the HRS60 family of repetitive DNA sequences can be induced with 5-azacytidine (5-azaC) in tobacco tissue cultures. We have also proven that such a DNA methylation status is maintained during the recovery of protoplasts, plant regeneration, and vegetative development. In the present paper we follow meiotic transmission of hypomethylated HRS60 DNA. Plants obtained from seeds treated with 5-azaC were either self pollinated or crossed with a non-treated control in a reciprocal way. Analysis of the methylation status of the HRS60 DNA revealed that these sequences were hypomethylated in the progenies up to the extent found in the parental 5-azaC-treated plant. Since no parent-of-origin effect was observed, we presume that both male and female gametes transmit an artificial methylation imprint to a similar extent. This result is supported by methylcytosine evaluation in the total genomic DNA samples. A temporal analysis of 5-azaC effects on germinating seeds and a phenotypic evaluation of 5-azaC-treated tobacco plants are also presented.

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References

  • An G (1994) Regulatory genes controlling flowering time or floral organ development. Plant Mol Biol 25:335–337

    Google Scholar 

  • Bezděk M, Koukalová B, Brzobohatý B, Vyskot B (1991) 5-azacytidine-induced hypomethylation of tobacco HRS60 tandem DNA repeats in tissue culture. Planta 184:487–490

    Google Scholar 

  • Bezděk M, Koukalová B, Kuhrová V, Vyskot B (1992) Differential sensitivity of CG and CCG DNA sequences to ethionine-induced hypomethylation of the Nicotiana tabacum genome. FEBS Lett 300:268–270

    Google Scholar 

  • Burn JE, Bagnall DJ, Metzger JD, Dennis ES, Peacock WJ (1993) DNA methylation, vernalization, and the initiation of flowering. Proc Natl Acad Sci USA 90:287–291

    Google Scholar 

  • Cedar H, Solage A, Glaser G, Razin A (1979) Direct detection of methylated cytosine in DNA by use of the restriction enzyme MspI. Nucleic Acids Res 6:2125–2132

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    CAS  Google Scholar 

  • Driscoll DJ (1994) Genomic imprinting in humans. In: Friedmann T (ed) Molecular genetic medicine, vol 4. Academic Press, San Diego, pp 37–77

    Google Scholar 

  • Fajkus J, Reich J (1991) Evaluation of restriction endonuclease cleavage of plant nuclear DNA using contaminating chloroplast DNA. Folia Biol (Prague) 37:224–226

    Google Scholar 

  • Fajkus J, Vyskot B, Bezděk M (1992) Changes in chromatin structure due to hypomethylation induced with azacytidine or DL-ethionine. FEBS Lett 314:13–16

    Google Scholar 

  • Fieldes MA (1993) Heritable effects of 5-azacytidine treatments on the growth and development of flax (Linum unsitatissimum) genotrophs and genotypes. Genome 37:1–11

    Google Scholar 

  • Flavell RB (1994) Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci USA 91:3490–3496

    CAS  PubMed  Google Scholar 

  • Follmann H, Balzer H-J, Schleicher R. (1990) Biosynthesis and distribution of methylcytosine in wheat DNA. How different are plant DNA methyltransferases? In: Clawson GA, Willis DB, Weissbach A, Jones PA (eds) Nucleic acid methylation. AR Liss Inc, New York, pp 199–210

    Google Scholar 

  • Hepburn AG, Clarke LE, Pearson L, White J (1983) The role of cytosine methylation in the control of nopaline synthase gene expression in a plant tumor. J Mol Appl Genet 2:315–329

    Google Scholar 

  • Jorgensen R (1993) The germinal inheritance of epigenetic information in plants. Phil Trans R Soc Lond B 339:173–181

    Google Scholar 

  • Kenton A, Parokonny AS, Gleba YY, Bennet MD (1993) Characterization of the Nicotiana tabacum genome by molecular cytogenetics. Mol Gen Genet 240:159–169

    Google Scholar 

  • Kermicle JL, Alleman M (1990) Gametic imprinting in maize in relation to the angiosperm life cycle. Development 1990 (Suppl):9–14

  • Kovkalová B, Reich J, Matyášek R, Kuhrová V, Bezděk M (1989) A BamHI family of highly repeated DNA sequences of Nicotiana tabacum. Theor Appl Genet 78:77–80

    Google Scholar 

  • Koukalová B, Kuhrová V, Vyskot B, Široký J, Bezděk M (1994) Maintenance of induced hypomethylated state of tobacco nuclear repetitive DNA sequences in the course of protoplast and plant regeneration. Planta 194:306–310

    Google Scholar 

  • Koařík A, Kukalová B, Holý A, Bezděk M (1994) Sequence-specific hypomethylation of tobacco genome induced with dihydroxypropyladenine, ethionine and 5-azacytidine. FEBS Lett 353:309–311

    Google Scholar 

  • Matzkle M, Matzke AJM (1993) Genomic imprinting in plants: parental effects and trans-inactivation phenomena. Annu Rev Plant Physiol Plant Mol Biol 44:53–76

    Google Scholar 

  • Ngernprasirtsiri J, Akazawa T (1990) Modulation of DNA methylation and gene expression in cultured sycamore cells treated by hypomethylating base analog. Eur J Biochem 194:513–520

    Google Scholar 

  • Phillips RL, Kaeppler SM, Olhoft P (1994) Genetic instability of plant tissue culture: breakdown of normal controls. Proc Natl Acad Sci USA 91:5222–5226

    CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sano H, Kamada I, Youssefian S, Wabiko H (989) Correlation between DNA undermethylation and dwarfism in maize. Biochem Biophys Acta 1009:35–38

  • Sano H, Kamada I, Youssefian S, Katsumi M, Wabiko H (1990) A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol Gen Genet 220:441–447

    Google Scholar 

  • Santi DV, Norment A, Garrett CE (1984) Covalent bond formation between a DNA-cytosine methyltransferase and DNA containing 5-azacytosine. Proc Natl Acad Sci USA 81:6993–6997

    Google Scholar 

  • Široký J, Janoušek B, Mouras A, Vyskot B (1994) Replication pattern of sex chromosomes in Melandrium album female cells. Hereditas 120:175–181

    Google Scholar 

  • Surani MA, Reik W, Allen ND (1988) Transgenes as molecular probes for genomic imprinting. Trends Genet 4:59–62

    Google Scholar 

  • Vyskot B, Fajkus J, Kuglík P, Koukalová B, Kuhrová V (1991) Genome modifications in protoplast-derived tobacco plants: phenotypic evaluation and RFLP analysis. Biol Plant (Prague) 33:455–460

    Google Scholar 

  • Vyskot B, Janoušek B, Široký J (1995) Epigenetic control of sex expression in a dioecious plant, Melandrium album. Genetics, (in press)

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Communicated by R. Hagemann

On leave from the School of Biology and Biochemistry, University of Bath, England

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Vyskot, B., Koukalová, B., Kovařík, A. et al. Meiotic transmission of a hypomethylated repetitive DNA family in tobacco. Theoret. Appl. Genetics 91, 659–664 (1995). https://doi.org/10.1007/BF00223294

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  • DOI: https://doi.org/10.1007/BF00223294

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