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.
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
Botta R, Vergano G, Me G, Vallania R (1995) Floral biology and embryo development in chestnut (Castanea sativa Mill.). Hort Sci 30:1283–1286
Carraway DT, Wilde HD, Merkle SA (1994) Somatic embryogenesis and gene transfer in American chestnut. Am Chestnut Found 8:29–33
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
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
Fraga M, Cañal MJ, Rodríguez R (2002a) Phase-change related epigenetic and physiological changes in Pinus radiata D. Don Planta 215:672–678
Fraga M, Cañal MJ, Rodríguez R (2002b) Genomic DNA methylation—demethylation during aging and reinvigoration of Pinus radiata. Tree Physiol 22:813–816
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
Giovannelli A, Giannini R (2000) Reinvigoration of mature chestnut (Castanea sativa) by repeated graftings and micropropagation. Tree Physiol 20:1243–1248
Hasbún R, Valledor L, Santamaría E, Cañal MJ, Rodríguez R (2007) Dynamics of DNA methylation in chestnut trees development. Acta Hort 760:563–566
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
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
Lippman Z, Martienssen R (2004) The role of RNA interference in heterochromatic silencing. Nature 431:364–370
McKay JW (1942) Self-sterility in the Chinese chestnut (Castanea mollisima). Proc Am Hort Sci 36:293–298
McKay JW, Crane HL (1938) The immediate effect of pollen on the fruit of the chestnut. Proc Amer Soc Hort Sci 41:156–160
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
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assay with tobacco tissue culture. Physiol Plant 15:473–497
Nakamura M (2001) Pollen tube growth and fertilization in Japanese chestnut (Castanea crenata Sieb. et Zucc.). J Jpn Soc Hort Sci 70:561–566
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
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
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
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
Sutherland S (1987) Why hermaphroditic plants produce many more flowers than fruits: experimental tests with Agave mckelveyana. Evolution 41:750–759
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
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
Zluvova J, Janousek B, Vyskot B (2001) Immunohistochemical study of DNA methylation dynamics during plant development. J Exp Bot 365:2265–2273
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by José Feijó.
Rights 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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00497-010-0145-9