Transcripts that specifically accumulate in transgenic tobacco plants expressing an anti-sense construct for a tobacco type I DNA methyltransferase, NtMET1, were screened by the differential display method. Of the 31 genes identified, 16 encoded proteins with known functions; ten of these were related to biotic and abiotic stress responses, and the other six to cellular functions. In order to examine whether expression of these genes is correlated with DNA methylation status under natural stress conditions, a pathogen-responsive gene ( NtAlix1) was selected as representative, and assayed for transcript induction and genomic methylation in tobacco plants infected with tobacco mosaic virus (TMV). In inoculated leaves of wild-type plants, NtAlix1 transcripts began to accumulate 12 h after the onset of the hypersensitive response (HR), and levels remained high for up to 24 h. Changes in the methylation status at the locus became obvious 24 h later, as detected by digestion of genomic DNA with a methylation-sensitive restriction enzyme. The results suggest that the level of DNA methylation may change in response to external stresses, and that this is closely related to the activation of stress-responsive genes.
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The authors are grateful to Dr. M. Bezdek (Academy of Sciences of the Czech Republic) for providing the plasmid HRS60.1, Drs N. Koizumi and Y. Yamaguchi (Nara Institute of Science and Technology) for valuable discussion and suggestions, to Ms M. Kobayashi (Nara Institute of Science and Technology) for technical assistance and to Dr M. Moore (Intermal, Nagoya) for critical reading of the manuscript. This work was supported by a grant from the Research for the Future Program (JSPS-RTFT001604) of the Japan Society for the Promotion of Science
Communicated by G. Jürgens
The first two authors contributed equally to this work
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Wada, Y., Miyamoto, K., Kusano, T. et al. Association between up-regulation of stress-responsive genes and hypomethylation of genomic DNA in tobacco plants. Mol Genet Genomics 271, 658–666 (2004). https://doi.org/10.1007/s00438-004-1018-4
- DNA methyltransferase
- Tobacco mosaic virus
- Hypersensitive response
- Nicotiana tabacum.