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Changes in DNA methylation during somatic embryogenesis in Cucurbita pepo L.

  • Cell Biology and Morphogenesis
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Abstract

Three pumpkin embryogenic lines were initiated on wounded zygotic embryos cultured on medium with or without 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryo development was controlled by the availability of various compounds in the medium: presence/absence of 2,4-D, nitrogen sources. The highest rate of DNA methylation was in the early embryo stages, predominantly on MSC medium with 2,4-D and on auxin-free medium supplemented with 1.0 mM NH4Cl. DNA methylation was correlated with early embryo development in a manner that was not exclusively dependent on the presence/absence of exogenous auxin. DNA methylation decreased during embryo maturation on auxin-free MSC medium and on auxin-free MSC supplemented with 12.3 μM 5-azacytidine (5-azaC). The embryogenic features of the pumpkin tissue were preserved, even after a 2-month treatment with 5-azaC.

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Fig. 1
Fig. 2a–d
Fig. 3
Fig. 4a–c

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Abbreviations

5-azaC:

5-Azacytidine

CRED-RA:

Coupled restriction enzyme digestion and random amplification

2,4-D:

2,4-Dichlorophenoxyacetic acid

DNMRT:

Duncan’s new multiple range test

IAA:

Indole-3-acetic acid

5-mC:

5-Methylcytosine

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Acknowledgements

We thank Ms. Ana-Marija Boljkovac for her technical assistance.

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Authors and Affiliations

  1. Department of Molecular Biology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000, Zagreb, Croatia

    Dunja Leljak-Levanić, Nataša Bauer & Sibila Jelaska

  2. Institute Ruder Boťkovic, Bijenicka 54, 10000, Zagreb, Croatia

    Snježana Mihaljević

Authors
  1. Dunja Leljak-Levanić
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  2. Nataša Bauer
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  3. Snježana Mihaljević
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  4. Sibila Jelaska
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Correspondence to Dunja Leljak-Levanić.

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Communicated by H. Lörz

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Leljak-Levanić, D., Bauer, N., Mihaljević, S. et al. Changes in DNA methylation during somatic embryogenesis in Cucurbita pepo L.. Plant Cell Rep 23, 120–127 (2004). https://doi.org/10.1007/s00299-004-0819-6

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  • DOI: https://doi.org/10.1007/s00299-004-0819-6

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