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Epigenetics of long-term somatic embryogenesis in Theobroma cacao L.: DNA methylation and recovery of embryogenic potential

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Abstract

In Theobroma cacao L., declined embryogenic potential was observed in regenerated somatic embryos from long-term secondary somatic embryogenesis (SE). In order to explore the relationship between DNA methylation and the long-term secondary SE, the embryogenic potential and global DNA methylation levels in young (12 months-old), aged (36 months-old) and extra somatic embryogenesis (39 months-old) subjected to different 5-Azacytidine (5-azaC) treatments were comparatively assessed. Global DNA methylation levels increased in aged somatic embryos with long-term in vitro culture, but 5-azaC-supplemented treatments resulted in unaltered levels. In addition, DNA methylation pattern during SE was not affected by 5-azaC. DNA methylation increased during SE expression. Interestingly, the extra SE induction showed that aged somatic embryos can recovery the embryogenic potential in treatment supplemented with 5-azaC at specific concentration. The outcome of this study suggested that the long-term SE in cacao induced the decline on embryogenic potential, which can be reversible trough 5-azaC supplementation. Besides, increased DNA methylation levels might be a response to the stress conditions that plant cells were exposed to during SE.

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Acknowledgements

This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Proc. 478393/2013-0, and 306126/2013-3), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação de Amparo a Pesquisa e Inovação do Estado de Santa Catarina (FAPESC, Proc. 2780/2012-4). The authors thank to Central Laboratory of Electron Microscopy (LCME) of the Federal University of Santa Catarina, Brazil.

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Authors and Affiliations

  1. Laboratório de Fisiologia do Desenvolvimento e Genética Vegetal, Centro de Ciências Agrárias, Universidade Federal de Santa Catarina, Florianópolis, SC, 88034-001, Brazil

    Liliana Alexandra Pila Quinga, Hugo Pacheco de Freitas Fraga, Leila do Nascimento Vieira & Miguel Pedro Guerra

  2. Laboratório de Micropropagação Vegetal, Departamento de Botânica, Universidade Federal do Paraná, Curitiba, PR, 81531-970, Brazil

    Hugo Pacheco de Freitas Fraga

  3. Tree and Timber Institute, National Research Council of Italy, Via Madonna del Piano 10, Sesto Fiorentino, 50019, Florence, Italy

    Miguel Pedro Guerra

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  1. Liliana Alexandra Pila Quinga
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  2. Hugo Pacheco de Freitas Fraga
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  3. Leila do Nascimento Vieira
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  4. Miguel Pedro Guerra
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Contributions

LAPQ and MPG: Conceived, designed and conducted the experiments. LAPQ and MPG: Data analysis and manuscript writing. HPFF and LNV: DNA methylation analysis and manuscript review. MPG: Contributed reagents/materials/analysis tools.

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Correspondence to Miguel Pedro Guerra.

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Communicated by M. Angeles Revilla.

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Pila Quinga, L.A., Pacheco de Freitas Fraga, H., do Nascimento Vieira, L. et al. Epigenetics of long-term somatic embryogenesis in Theobroma cacao L.: DNA methylation and recovery of embryogenic potential. Plant Cell Tiss Organ Cult 131, 295–305 (2017). https://doi.org/10.1007/s11240-017-1284-6

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