Skip to main content
Log in

DNA methylation during sexual embryogenesis and implications on the induction of somatic embryogenesis in Castanea sativa Miller

  • Original Article
  • Published:
Sexual Plant Reproduction Aims and scope Submit manuscript

Abstract

From anthesis to mature seed formation, burrs from cross-pollinated adult Castanea sativa Miller trees were characterized and seven developmental stages defined based on macro and micromorphological traits. In order to get an insight into the involvement of epigenetic mechanisms in sexual embryogenesis and to define somatic embryogenesis induction capability, global DNA methylation and the somatic embryogenic competence were quantified. On cross-pollinated trees once fertilization takes place, at least one ovule per ovary becomes dominant, and transient DNA demethylation occurs coinciding with the start of the sexual embryogenic programme. Unfertilized ovules from the same cluster, which maintain their prior size, increase their methylation level and undergo degeneration. These results were validated using non-cross-pollinated trees and the asynchrony of flower receptivity. When testing in vitro somatic embryogenesis response of isolated dominant ovules and axes from zygotic embryos under cross-pollinated conditions, the highest competence was found for reaching seed maturity. Thus, a “developmental window” of somatic embryogenesis in chestnut has been characterized. It includes from fertilization to embryo maturity, and a transient decrease in methylation is necessary after fertilization for the development of the somatic embryogenesis response.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Baurens FC, Nicolleau J, Legavre T, Verdeil JL, Monteuuis O (2004) Genomic DNA methylation of juvenile and mature Acacia mangium micropropagated in vitro with reference to leaf morphology as a phase change marker. Tree Physiol 24:401–407

    CAS  PubMed  Google Scholar 

  • Botta R, Vergano G, Me G, Vallania R (1995) Floral biology and embryo development in chestnut (Castanea sativa Mill.). Hort Sci 30:1283–1286

    Google Scholar 

  • Carraway DT, Wilde HD, Merkle SA (1994) Somatic embryogenesis and gene transfer in American chestnut. Am Chestnut Found 8:29–33

    Google Scholar 

  • Chakrabarty D, Yu KW, Paek KY (2003) Detection of DNA methylation changes during somatic embryogenesis of Siberian ginseng (Eleuterococcus senticosus). Plant Sci 165:61–68

    Article  CAS  Google Scholar 

  • Finnegan EJ, Peacock WJ, Dennis ES (1996) Reduced DNA methylation in Arabidopsis thaliana results in abnormal plant development. Proc Natl Acad Sci USA 93:8449–8454

    Article  CAS  PubMed  Google Scholar 

  • Fraga M, Cañal MJ, Rodríguez R (2002a) Phase-change related epigenetic and physiological changes in Pinus radiata D. Don Planta 215:672–678

    Article  CAS  Google Scholar 

  • Fraga M, Cañal MJ, Rodríguez R (2002b) Genomic DNA methylation—demethylation during aging and reinvigoration of Pinus radiata. Tree Physiol 22:813–816

    CAS  PubMed  Google Scholar 

  • Fraga M, Uriol E, Diego LB, Berdasco M, Esteller M, Cañal MJ, Rodríguez R (2002c) High performance capillary electrophoretic method for the quantification of 5-methyl 2′-deoxycytidine in genomic DNA: application to plant, animal and human cancer tissues. Electrophoresis 23:1677–1681

    Article  CAS  PubMed  Google Scholar 

  • Giovannelli A, Giannini R (2000) Reinvigoration of mature chestnut (Castanea sativa) by repeated graftings and micropropagation. Tree Physiol 20:1243–1248

    PubMed  Google Scholar 

  • Hasbún R, Valledor L, Santamaría E, Cañal MJ, Rodríguez R (2007) Dynamics of DNA methylation in chestnut trees development. Acta Hort 760:563–566

    Google Scholar 

  • Hasbún R, Valledor L, Rodríguez JL, Santamaría E, Ríos D, Sánchez M, Cañal MJ, Rodríguez R (2008) HPCE quantification of 5-methyl-20-deoxycytidine in genomic DNA: Methodological optimization for chestnut and other woody species. Plant Physiol Biochem 20:1–8

    Google Scholar 

  • Ikegami K, Ohgane J, Tanaka S, Yagi S, Shiota K (2009) Interplay between DNA methylation, histone modification and chromatin remodelling in stem cells and during development. Int J Dev Biol 53:203–214

    Article  CAS  PubMed  Google Scholar 

  • Lippman Z, Martienssen R (2004) The role of RNA interference in heterochromatic silencing. Nature 431:364–370

    Article  CAS  PubMed  Google Scholar 

  • McKay JW (1942) Self-sterility in the Chinese chestnut (Castanea mollisima). Proc Am Hort Sci 36:293–298

    Google Scholar 

  • McKay JW, Crane HL (1938) The immediate effect of pollen on the fruit of the chestnut. Proc Amer Soc Hort Sci 41:156–160

    Google Scholar 

  • Meijón M, Valledor L, Rodríguez JL, Hasbún R, Santamaría E, Feito I, Cañal MJ, Berdasco M, Fraga MF, Rodríguez R (2008) Plant epigenetics. In: Esteller Manel (ed) Epigenetics in biology and medicine. Elsevier, London, pp 225–239

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assay with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nakamura M (2001) Pollen tube growth and fertilization in Japanese chestnut (Castanea crenata Sieb. et Zucc.). J Jpn Soc Hort Sci 70:561–566

    Article  Google Scholar 

  • Peano C, Bounous G, Paglietta R (1990) Contributo allo studio della biologia fiorale e di fruttificazione di cultivar europee, orientali ed ibridi del genere Castanea Mill. Annali della Facoltà di Scienze Agrarie della Università degli Studi di Torino 16:83–89

    Google Scholar 

  • Şan B, Sezgin M, Dumanoğlu KöksalI (2007) Somatic embryogenesis from immature cotyledons of some European Chestnut (Castanea sativa Mill.) cultivars. Turk J Agric For 31:175–179

    Google Scholar 

  • Santamaría E, Hasbún R, Valera MJ, Meijón M, Valledor L, Rodríguez JL, Toorop PE, Cañal MJ, Rodríguez R (2009) Acetylated H4 histone and genemic DNA methylation patterns during bud set and bud burst in Castanea sativa. J Plant Phy 166:1360–1369

    Article  Google Scholar 

  • Sauer U, Wilhelm E (2005) Somatic embryogenesis from ovaries, developing ovules and immature zygotic embryos, and improved embryo development of Castanea sativa. Biol Plant 49:1–6

    Article  Google Scholar 

  • Sutherland S (1987) Why hermaphroditic plants produce many more flowers than fruits: experimental tests with Agave mckelveyana. Evolution 41:750–759

    Article  Google Scholar 

  • Valledor L, Hasbún R, Meijón M, Rodríguez JL, Santamaría E, Viejo M, Berdasco M, Feito I, Fraga MF, Cañal MJ, Rodríguez R (2007) Involvement of DNA methylation in tree development and micropropagation. Plant Cell Tiss Organ Cult 91:75–86

    Article  CAS  Google Scholar 

  • Valledor L, Meijón M, Hasbún R, Cañal MJ, Rodríguez R (2009) Variations in DNA methylation, acetylated histone H4, and methylated histone H3 during Pinus radiata needle maturation in relation to the loss of in vitro organogenic capability. J Plant Phy 167:351–357

    Google Scholar 

  • Zluvova J, Janousek B, Vyskot B (2001) Immunohistochemical study of DNA methylation dynamics during plant development. J Exp Bot 365:2265–2273

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M. J. Cañal.

Additional information

Communicated by José Feijó.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Viejo, M., Rodríguez, R., Valledor, L. et al. DNA methylation during sexual embryogenesis and implications on the induction of somatic embryogenesis in Castanea sativa Miller. Sex Plant Reprod 23, 315–323 (2010). https://doi.org/10.1007/s00497-010-0145-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00497-010-0145-9

Keywords

Navigation