Abstract
The effect of benzyladenine (BA) on the activation of rRNA gene transcription was studied in wheat (Triticum aestivum and T. urartu) as related to the phenomenon of nucleolar dominance and the changes in the extent of methylation of the intergenic spacers in the subgenome A. The method of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze the fragments of rDNA promoter regions amplified with the primers designed to recognize the sites of DNA isolated from BA-treated seedlings of diploid T. urartu and hexaploid T. aestivum and desaminated with metabisulfite. The subsequent genomic bisulfite sequencing of the amplification products was used to evaluate the level of methylation/demethylation of the particular cytosine residues. BA diminished methylation of cytosine residues in rDNA promoter regions to the level, which was different in two wheat species; these data presume that the transcriptional activities of rRNA genes in various wheat subgenomes depend on the extent of their methylation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BA:
-
benzyladenine
- IGS:
-
intergenic spacer
- NOR:
-
nucleolar organizer
- PCR:
-
polymerase chain reaction
- SSCP:
-
single strand conformation polymorphism
- TIS:
-
transcription initiation site
References
Karpov, V.L., DNA, Chromatin, Histone Code, Vestn. Ross. Akad. Nauk, 2003, vol. 73, pp. 505–513.
Bird, A., DNA Methylation Patterns and Epigenetic Memory, Gene Dev., 2002, vol. 16, pp. 6–21.
Vanyushin, B.F., Enzymatic DNA Methylation — Epigenetic Control of Genetic Functions in the Cell, Biokhimiya, 2005, vol. 70, pp. 598–611.
Vanyushin, B.F., DNA Methylation in Plants, Curr. Top. Microbiol. Immunol., 2006, vol. 301, pp. 67–122.
Finnegan, E.J., Genger, R.K., Peacock, W.J., and Dennis, E.S., DNA Methylation in Plants, Annu. Rev. Plant Physiol. Plant Mol. Biol., 1998, vol. 49, pp. 223–247.
Gruenbaum, I., Naveh-Many, T., Cedar, H., and Razin, A., Sequence Specifity of Methylation in Higher Plant DNA, Nature, 1981, vol. 292, pp. 860–862.
Babinger, P., Kobl, I., Mages, W., and Schmitt, R., A Link between DNA Methylation and Epigenetic Silencing in Transgenic Volvox carteri, Nucleic Acids Res., 2001, vol. 29, pp. 1261–1271.
Baylin, S.B. and Herman, J.G., DNA Hypermethylation in Tumorigenesis: Epigenetics Joins Genetics, Trends Genet., 2000, vol. 16, pp. 168–174.
Chang, S. and Pikaard, C.S., Transcript Profiling in Arabidopsis Reveals Complex Responses to Global Inhibition of DNA Methylation and Histone Deacetylation, J. Biol. Chem., 2005, vol. 280, pp. 796–804.
Houchins, K., O’Dell, M., Flavell, R.B., and Gustafson, J.P., Cytosine Methylation and Nucleolar Dominance in Cereal Hybrids, Mol. Gen. Genet., 1997, vol. 255, pp. 294–301.
Thompson, W.F. and Flavell, R.B., DNase I Sensitivity of Ribosomal RNA Genes in Chromatin and Nucleolar Dominance in Wheat, J. Mol. Biol., 1988, vol. 204, pp. 535–548.
Vakhitov, V.A., Chemeris, A.V., Sabirzhanov, B.E., Akhunov, E.D., Kulikov, A.M., Nikonorov, Yu.M., Gimalov, F.R., Bikbulatova, S.M., and Baimiev, Al.Kh., Investigation of Triticum L. and Aegilops L. Phylogeny by Comparison of Nucleotide Sequences in rDNA Promoter Regions, Genetika, 2003, vol. 39, pp. 5–17.
Akhunov, E.D., Chemeris, A.V., Kulikov, A.V., and Vakhitov, V.A., Functional Analysis of Diploid Wheat rRNA Promoter, Biochim. Biophys. Acta, 2001, vol. 1522, pp. 226–229.
Pikaard, C.S. and Chen, Z.J., Nucleolar Dominance, Transcription of Ribosomal RNA Genes by Eukaryotic RNA Polymerase I, Paule, M.R., Ed., Geargetown, TX: R. G. Landes, 1998, pp. 277–294.
Fatkhutdinova, R.A., Chemeris, A.V., and Vakhitov, V.A., Some Features of Cytosine Residue Methylation in rDNA of Diploid Triticum urartu Thurm. and Diploid Aegilops umbellulata Zhuk., Genetika, 1998, vol. 34, pp. 512–519.
Vlasova, T.I., Kirnos, M.D., Demidenko, Z.N., and Vanyushin, B.F., Phytohormone-Induced In Vitro Modulation of Methylation in Nucleolar Wheat DNA Using Wheat Cytosine DNA-Methyl Transferase, Biokhimiya, 1994, vol. 59, pp. 1872–1881.
Gaudino, R.J. and Pikaard, C.S., Cytokinin Induction of RNA Polymerase I Transcription in Arabidopsis thaliana, J. Biol. Chem., 1997, vol. 272, pp. 6799–6804.
Frommer, M., McDonald, L.E., Millar, D.S., Collis, C.M., Watt, F., Grigg, G.W., Molloy, P.L., and Paul, C.L., A Genomic Sequencing Protocol That Yields a Positive Display of 5-Methylcytosine Residues in Individual DNA Strands, Proc. Natl. Acad. Sci. USA, 1992, vol. 89, pp. 1827–1831.
Cedar, H., Solage, A., Glaser, G., and Razin, A., Direct Detection of Methylated Cytosine in DNA by Use of the Restriction Enzyme MspI, Nucleic Acids Res., 1979, vol. 6, pp. 2125–2132.
Koukalova, B., Fojtova, M., Lim, K.Y., Fulnecek, J., Leitch, A.R., and Kovarik, A., Dedifferentiation of Tobacco Cells Is Associated with Ribosomal RNA Gene Hypomethylation, Increased Transcription, and Chromatin Alterations, Plant Physiol., 2005, vol. 139, pp. 275–286.
D’yachenko, O.V., Zakharchenko, N.S., Shevchuk, T.V., Bonert, Kh., Kushman, Dzh., and Bur’yanov, Ya.I., Hypermethylation of CCWGG-Sequences in Mesembryanthemum crystallinum DNA during Their Adaptation to Salt Stress, Biokhimiya, 2006, vol. 71, pp. 570–575.
Komarova, N.Y., Grabe, T., Hemleben, V., Huigen, D.J., and Volkov, R.A., Organization, Differential Expression and Methylation of rDNA in Artificial Solanum Allopolyploids, Plant Mol. Biol., 2004, vol. 56, pp. 439–463.
Graham, D.E., The Isolation of High Molecular Weight DNA from Whole Organisms or Large Tissue Masses, Anal. Biochem., 1978, vol. 85, pp. 609–613.
Raizis, A.M., Schmitt, F., and Jost, J.P., A Bisulfite Method of 5-Methylcytosine Mapping That Minimized Template Degradation, Anal. Biochem., 1995, vol. 226, pp. 161–166.
Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular, Cloning. A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab., 1989.
Kinoshita, H., Shi, Y., Sandefur, C., and Jarrard, D., Screening Hypermethylated Regions by Methylation-Sensitive Single-Strand Conformation Polymorphism, Anal. Biochem., 2000, vol. 278, pp. 165–169.
Maxam, A.M. and Gilbert, W., Sequencing End-Labeled DNA with Base-Specific Chemical Cleavage, Methods Enzymol., 1980, vol. 65, pp. 499–560.
Cohen, S.N., Chang, A.C.Y., and Hsu, L., Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of E. coli by R-Factor DNA, Proc. Natl. Acad. Sci. USA, 1972, vol. 69, pp. 2110–2114.
Sanger, F., Nicklen, S., and Coulson, A.R., DNA Sequencing with Chain-Termination Inhibitors, Proc. Natl. Acad. Sci. USA, 1977, vol. 74, pp. 5463–5467.
Fatkhutdinova, R.A., Shakirova, F.M., Chemeris, A.V., Sabirzhanov, B.E., and Vakhitov, V.A., NOR Activity in Wheat Species with Different Ploidy Levels Treated with Phytohormones, Genetika, 2002, vol. 38, pp. 1575–1579.
Lawrence, R.J., Earley, K., Pontes, O., Silva, M., Chen, Z.J., Neves, N., Viegas, W., and Pikaard, C.S., A Concerted DNA Methylation/Histone Methylation Switch Regulates rRNA Gene Dosage Control and Nucleolar Dominance, Mol. Cell, 2004, vol. 13, pp. 599–609.
Author information
Authors and Affiliations
Additional information
Original Russian Text © B.E. Sabirzhanov, S.M. Bikbulatova, R.A. Fatkhutdinova, A.V. Chemeris, F.M. Shakirova, V.A. Vakhitov, 2007, published in Fiziologiya Rastenii, 2007, Vol. 54, No. 2, pp. 235–242.
Rights and permissions
About this article
Cite this article
Sabirzhanov, B.E., Bikbulatova, S.M., Fatkhutdinova, R.A. et al. Effect of benzyladenine on the extent of methylation of cytosine residues of the intergenic spacers in rDNA promoter regions of Triticum aestivum and Triticum urartu . Russ J Plant Physiol 54, 207–214 (2007). https://doi.org/10.1134/S1021443707020082
Received:
Issue Date:
DOI: https://doi.org/10.1134/S1021443707020082