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Arabidopsis Displays Centromeric DNA Hypomethylation and Cytological Alterations of Heterochromatin Upon Attack by Pseudomonas syringae

June 2006 , Volume 19 , Number  6
Pages  577 - 587

Valeria Pavet , 1 Cristián Quintero , 1 Nicolás M. Cecchini , 1 Alberto L. Rosa , 2 and María E. Alvarez 1

1CIQUIBIC—Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, 5000 Córdoba, Argentina; 2Instituto de Investigación Médica Mercedes y Martín Ferreyra—INIMEC, Friuli 2434, 5016 Córdoba, Argentina

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Accepted 25 January 2006.

Plant tissues display major alterations upon the perception of microbial pathogens. Changes of cytoplasmic and apo-plastic components that sense and transduce plant defenses have been extensively characterized. In contrast, less information is available about modifications affecting the plant nuclear genome under these circumstances. Here, we investigated whether the Arabidopsis thaliana DNA methylation status is altered in tissues responding to the attack of Pseudomonas syringae pv. tomato DC3000. We applied amplified fragment length polymorphism analysis to monitor cytosine methylation at anonymous 5′-CCGG-3′ and 5′-GATC-3′ sites in naïve and infected samples. Plant genomic fragments reducing methylation upon infection, including peri/centromeric repeats such as the 180-bp unit, Athila retrotansposon, and a portion of the nuclear insertion of mitochondrial DNA, were isolated and characterized. P. syringae pv. tomato-induced hypomethylation was detected by high-performance liquid chromatography assays and at the molecular level it did not seem to equally affect all 5-methyl cytosine (5-mC) residues. Nuclei from challenged tissues displayed structural chromatin alterations, including loosening of chromocenters, which also were stimulated by avirulent P. syringae pv. tomato, but not by the P. syringae pv. tomato hrpL¯ mutant. Finally, P. syringae pv. tomato-induced hypomethylation was found to occur in the absence of DNA replication, suggesting that it involves an active demethylation mechanism. All these responses occurred at 1 day postinfection, largely preceding massive plant cell death generated by pathogen attack.

Additional keywords: biotic stress, DNA methylation changes, interphase nuclei.

© 2006 The American Phytopathological Society