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Analysis of genomic DNA methylation patterns in regenerated and control plants of rye (Secale cereale L.)

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Abstract

Rye (Secale cereale L.) is a species that has shown high rates of somaclonal variation when plants obtained by in vitro culture were analysed using different techniques. In this study, using methylation-sensitive amplified polymorphism (MSAP) markers, we analysed the cytosine methylation status at genomic level of regenerated plants of rye that were obtained by somatic embryogenesis. Such plants were originated from three different cell lines and the results were compared with the data obtained from the control plants grown from seeds of the same cultivar and lot. A similar total number of MSAP markers was observed in the regenerated (937) and control plants (1,022), while the mean number detected per plant was significantly higher in regenerated (554.43) than in control plants (356.00). The analysis indicated conservation of the number of partially-methylated CCGG/GGCC sites for all type of plants. However the mean number of non-methylated sites was near twofold in the regenerated plants (442.48) than in control plants (248.19). Methylation changes have been detected in all the regenerated plants when compared within cell lines, with an average frequency of 9.01 % of the detected markers. We also observed that regenerated plants from one or several cell lines shared methylation changes at the same locus pointing to a non-random behaviour of the changes in genomic methylation.

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Acknowledgments

First, we thank the anonymous reviewers for their valuable comments that helped to considerably improve the quality of the manuscript. This work was supported by the Spanish DGICYT grant AGL2006-14249-C02-02 and by the Junta de Castilla y León grant LE052A06.

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  1. Área de Genética, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, 24071, León, Spain

    A. I. González, A. Sáiz, A. Acedo, M. L. Ruiz & C. Polanco

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  1. A. I. González
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  2. A. Sáiz
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  3. A. Acedo
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  4. M. L. Ruiz
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Correspondence to C. Polanco.

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González, A.I., Sáiz, A., Acedo, A. et al. Analysis of genomic DNA methylation patterns in regenerated and control plants of rye (Secale cereale L.). Plant Growth Regul 70, 227–236 (2013). https://doi.org/10.1007/s10725-013-9794-7

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