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Global DNA methylation profiles of somatic embryos of peach palm (Bactris gasipaes Kunth) are influenced by cryoprotectants and droplet-vitrification cryopreservation

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Abstract

Regrowth capacity and genetic stability of plants recovered following cryopreservation are associated with changes in DNA epigenetics, particularly in DNA methylation levels. In this study, global DNA methylation profiles associated with frequency of regrowth of peach palm (Bactris gasipaes) somatic embryos following cryopreservation using droplet-vitrification were investigated. Somatic embryo clusters (SEC) subjected to plant vitrification solution 3 (PVS3) for different durations (0, 60, 120, 180, and 240 min) were evaluated for regrowth capacity. The highest frequency of regrowth (52.4 %) was obtained when SEC were incubated in PVS3 for 120 min prior to droplet-vitrification cryopreservation. Global DNA methylation profiles were influenced by both cryoprotectants and droplet-vitrification cryopreservation. Incubation of SEC in PVS3 for limited durations not only reduced frequency of regrowth, but also increased DNA methylations levels when compared with proliferating SEC grown in a temporary immersion system. Although SEC subjected to cryopreservation exhibited the highest DNA methylation variation, 120 min SEC incubation in a PVS3 solution resulted in the recovery of initial global methylation profiles after 24 weeks of regrowth.

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Abbreviations

CIA:

Chloroform-isoamyl alcohol

CTAB:

Cetyltrimethyl ammonium bromide

DMSO:

Dimethylsulfoxide

Dntp:

Deoxynucleotide triphosphates

EDTA:

Ethylenediaminetetraacetic acid

FM:

Fresh mass

HPLC:

High-performance liquid chromatography

LN:

Liquid nitrogen

PVS:

Plant vitrification solution

SEC:

Somatic embryo clusters

SNK:

Student–Newman–Keuls

TIS:

Temporary immersion system

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Acknowledgments

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC) and Cordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Authors and Affiliations

  1. Graduate Program in Plant Genetic Resources, Plant Developmental Physiology and Genetics Laboratory, Federal University of Santa Catarina, Rod. Admar Gonzaga, Km 3, Florianópolis, SC, 88034-001, Brazil

    Angelo S. Heringer, Hugo P. F. Fraga, Leila N. Vieira, Joseph Francis Ree & Miguel P. Guerra

  2. Biosomatica Institute, Holambra, SP, 13825-000, Brazil

    Douglas A. Steinmacher

Authors
  1. Angelo S. Heringer
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  2. Douglas A. Steinmacher
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  3. Hugo P. F. Fraga
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  4. Leila N. Vieira
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  5. Joseph Francis Ree
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  6. Miguel P. Guerra
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Correspondence to Miguel P. Guerra.

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Heringer, A.S., Steinmacher, D.A., Fraga, H.P.F. et al. Global DNA methylation profiles of somatic embryos of peach palm (Bactris gasipaes Kunth) are influenced by cryoprotectants and droplet-vitrification cryopreservation. Plant Cell Tiss Organ Cult 114, 365–372 (2013). https://doi.org/10.1007/s11240-013-0331-1

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