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Analysis of Cytosine Methylation Pattern in Response to Water Deficit in Pea Root Tips
Authors:M. Labra  A. Ghiani  S. Citterio  S. Sgorbati  F. Sala  C. Vannini  M. Ruffini-Castiglione  M. Bracale
Affiliation:Dipartimento di Scienze dell 'Ambiente e del Territorio Universita' degli Studi di Milano-Bicocca, Milano, Italy;Dipartimento di Biologia, Universitàdi Milano, Via Celoria 26, 20133 Milano, Italy;DBSF, Universitàdell Insubria, Via J. H. Dunant 3, 21100 Varese, Italy;Istituto di Mutagenesi e Differenziamento, CNR Area della Ricerca, Via Moruzzi, 56124 Pisa, Italy
Abstract:Abstract: The correlation between environmental stress and DNA methylation has been studied by following the methylation status of cytosine residues in the DNA of pea root tips exposed to water deficit. DNA methylation was evaluated by two complementary approaches: (i) immunolabelling by means of a monoclonal antibody against 5-methylcytosine; (ii) MSAP (Methylation-Sensitive Amplified Polymorphism) to verify if methylation and de-methylation in response to water deficit may be related to specific DNA sequences. Immunolabelling showed that water stress induces cytosine hypermethylation in the pea genome. Regarding the CCGG target sequence, an increase in methylation specifically in the second cytosine (about 40 % of total site investigated) was revealed by MSAP analyses. In addition, MSAP band profile detected in three independent repetitions was highly reproducible suggesting that, at least for the CCGG target sequence, methylation was addressed to specific DNA sequences.
Keywords:Pisum sativum L    DNA methylation    water stress    MSAP
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