Skip to main content
SpringerLink
Log in

Methylation patterns revealed by MSAP profiling in genetically stable somatic embryogenic cultures of Ocotea catharinensis (Lauraceae)

  • Embryogenesis/somatic embryogenesis
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Ocotea catharinensis is a rare tree species indigenous to the Atlantic rainforest of South America. In spite of its value as a hardwood species, it is in danger of extinction. The species erratically produces seeds showing irregular flowering and slow growth. Therefore, plants are not easily replaced. Tissue culture-based techniques are commonly used for obtaining living material for tree propagation and in vitro preservation. Therefore, a high-frequency somatic embryogenic system was developed for the species. In the present work, the genetic fidelity of cell aggregates and somatic embryos at various stages of in vitro development of O. catharinensis was investigated using RAPD and AFLP markers. Both analyses confirmed the absence of genetic variation in all developmental stages of O. catharinensis embryogenic cultures, verifying that the in vitro system is genetically stable. The cultures were also analyzed for their methylation profiles at 5′-CCGG-3′ sites by identifying methylation-sensitive amplification polymorphisms. Some of these markers differentiated cell aggregates from embryo bodies. The sequencing of ten MSAP markers revealed that four sequences showed significant similarity to genes encoding plant proteins. Particularly, the predicted amino acid sequence of the fragment designated as OcEaggHMttc155 was similar to the enzyme 1–aminocyclopropane-1-carboxylate oxidase (ACO), which is involved in the biosynthesis of ethylene, and its expression was reported to occur from the beginning to the intermediate stages of plant embryo development. Here, we suggest that this enzyme is possibly involved in the control of the earliest stages of somatic embryogenesis of O. catharinensis, and an approach to study ACO expression during somatic embryogenesis is proposed.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.

Similar content being viewed by others

References

  • Altschul S. F.; Gish W.; Miller W.; Myers E. W.; Lipman D. J. Basic local alignment search tool. Journal of Molecular Biology 215: 403–410; 1990.

    PubMed  CAS  Google Scholar 

  • Bardini M.; Labra M.; Winfield M.; Sala F. Antibiotic-induced DNA methylation changes in calluses of Arabidopsis thaliana. Plant Cell, Tissue and Organ Culture 72: 157–162; 2003.

    Article  CAS  Google Scholar 

  • Bednarek P. T.; Orłowska R.; Koebner R. M. D.; Zimny J. Quantification of the tissueculture induced variation in barley (Hordeum vulgare L.). BMC Plant Biology 7: 10–19; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Carolan J. C. Using AFLP markers for species differentiation and assessment of genetic variability of in vitro-cultured Papaver bracteatum (section Oxytona). In Vitro Cellular & Developmental Biology. Plant 38: 300–307; 2007.

    Article  CAS  Google Scholar 

  • Chakrabaty D.; Yu K. W.; Paek K. Y. Detection of DNA methylation changes during somatic embryogenesis of Siberian ginseng (Eleuterococcus senticosus). Plant Science 165: 61–68; 2003.

    Article  CAS  Google Scholar 

  • Cottage A.; Yang A.; Maunders H.; Lacy C.; Ramsay N. Identification of DNA sequences flanking T-DNA insertions by PCR-walking. Plant Molecular Biology Reporter 19: 321–327; 2001.

    Article  CAS  Google Scholar 

  • Creste S.; Tulmann Neto A.; Figueira A. Detection of single sequence repeats polymorphims in denaturing polyacrylamide sequencing gels by silver staining. Plant Molecular Biology Reporter 19: 229–306; 2001.

    Article  Google Scholar 

  • Dalal N. V.; Rai R Vb. In vitro propagation of Ochreinauclea missionis (Wall. EX G. Don), an ethnomedicinal endemic and threatened tree. In Vitro Cell Dev Biol-Plant 37: 820–823; 2001.

    Article  CAS  Google Scholar 

  • Ferrándiz C. Regulation of fruit dehiscence in Arabidopsis. Journal of Experimental Botany 53: 2031–2038; 2002.

    Article  PubMed  Google Scholar 

  • Gagliardi R. F.; Hanai L. R.; Pacheco G.; Oliveira C. A.; Carneiro L. A.; Valls J. F. M.; Mansur E.; Vieira M. L. C. Assessment of genetic stability among in vitro plants of Arachis retusa using RAPD and AFLP markers for germplasm preservation. Journal of Integrative Plant Biology 49: 307–312; 2007.

    Article  CAS  Google Scholar 

  • Gómez-Jiménez M. C.; Matilla A. J.; Garrido D. Isolation and characterization of a cDNA encoding an ACC oxidase from Cicer arietinum and its expression during embryogenesis and seed germination. Australian Journal of Plant Physiology 25: 765–773; 1998.

    Article  Google Scholar 

  • Gruenbaum Y.; Naveh-Many T.; Cedar H.; Razin A. Sequence specificity of methylation in higher plant DNA. Nature 292: 860–862; 1981.

    Article  PubMed  CAS  Google Scholar 

  • Hall T. A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98; 1999.

    CAS  Google Scholar 

  • Hornero J.; Martinez I.; Celestino C.; Gallego F. J.; Torres V.; Toribio M. Early checking of genetic stability of cork oak somatic embryos by AFLP analysis. Integrative Journal of Plant Science 162: 827–833; 2001.

    Article  CAS  Google Scholar 

  • Joyce S. M.; Cassells A. C. Variation in potato microplant morphology in vitro and DNA methylation. Plant Cell, Tissue and Organ Culture 70: 125–137; 2002.

    Article  CAS  Google Scholar 

  • Kageyama P.; Reis A. Areas of secondary vegetation in the Itajai Valley Santa Catarina, Brazil. Perspectives for management and conservation. Forest Genetic Resources Information 21: 37–39; 1993.

    Google Scholar 

  • Leljak-Levanic D.; Bauer N.; Mihaljevic S.; Jelaska S. Changes in DNA methylation during somatic embryogenesis in Cucurbita pepo L. Plant Cell Reports 23: 120–127; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Lloyd G.; Mccown B. Commercially feasible micropropagation of mountain laurel, Kamila latifolia, by use of shoot tip culture. Proceedings of the International Plant Propagators Society 30: 421–427; 1980.

    Google Scholar 

  • Lo Schiavo F.; Pitto L.; Giuliano G.; Nuti-Ronchi V.; Marazziti D.; Vergera R.; Orselli S.; Terzi M. DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs. Theoretical and Applied Genetics 77: 325–331; 1989.

    Article  CAS  Google Scholar 

  • Lopes R.; Lopes M. T. G.; Carneiro M. S.; Matta F. P.; Camargo L. E. A.; Vieira M. L. C. Linkage and mapping of resistance genes to Xanthomonas axonopodis pv. passiflorae in yellow passion fruit. Genome 49: 17–29; 2006.

    Article  PubMed  CAS  Google Scholar 

  • Matilla A. J. Ethylene in seed formation and germination. Seed Science Research 10: 111–126; 2000.

    CAS  Google Scholar 

  • Matthes M.; Singh R.; Cheah S.-C.; Karp A. Variation in oil palm (Elaeis guineensis Jacq.) tissue culture-derived regenerants revealed by AFLPs with methylation-sensitive enzymes. Theoretical and Applied Genetics 102: 971–979; 2001.

    Article  CAS  Google Scholar 

  • Mcclelland M.; Nelson M.; Raschke E. Efect of site-specifc modifcation on restriction endonucleases and DNA modifcation methyltransferases. Nucleic Acids Research 22: 3640–3659; 1994.

    Article  PubMed  CAS  Google Scholar 

  • Moura-Costa P. H.; Viana A. M.; Mantell S. H. In vitro plantlet regeneration of Ocotea catharinensis Mez. (Lauraceae), an endangered forest tree of South Brazil. Plant Cell, Tissue and Organ Culture 35: 279–286; 1993.

    Article  Google Scholar 

  • Murashigue T.; Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15: 473–479; 1962.

    Article  Google Scholar 

  • Phillips R. L.; Kaeppler S. M.; Peschke V. M. Do we understand somaclonal variation? In: Nijkamp H. J. J.; Van Der Plas L. H. W.; Van Aartrijk J. (eds) Progress in Plant Cellular and Molecular Biology. Kluwer, Dordrecht, The Netherlands, pp 131–141; 1990.

    Google Scholar 

  • Renau-Morata B.; Nebauer S. G.; Arrillaga I.; Segura J. Assessments of somaclonal variation in micropropagated shoots of Cedrus: consequences of axillary bud breaking. Tree Genetics and Genomes 1: 3–10; 2005.

    Article  Google Scholar 

  • Reyna-López G. E.; Simpson J.; Ruiz-Herrera J. Differences in DNA methylation pattern are detectable during the dimorphic transition of fungi by amplification of restriction polymorphisms. Molecular & General Genetics 253: 703–710; 1997.

    Article  Google Scholar 

  • Rodríguez-Gacio M. C.; Nicols C.; Matilla A. J. The final step of the ethylene biosynthesis pathway in turnip tops (Brassica rapa L. cv. Rapa): Molecular characterization of the 1-aminocyclopropane-1-carboxylate oxidase BrACO1 throughout zygotic embryogenesis and germination of heterogeneous seeds. Physiologia Plantarum 121: 132–140; 2004.

    Article  Google Scholar 

  • Sanger F.; Nicklen S.; Coulson A. R. DNA sequencing with chain—terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74: 5463–5467; 1977.

    Article  PubMed  CAS  Google Scholar 

  • Santa-Catarina C.; Hanai L. R.; Viana A. M.; Dornelas M. C.; Floh E. I. S. SERK gene homolog expression, polyamines and amino acids associated with somatic embryogenic competence of Ocotea catharinensis Mez. (Lauraceae). Plant Cell, Tissue and Organ Culture 79: 53–61; 2004a.

    Article  CAS  Google Scholar 

  • Santa-Catarina C.; Olmedo A. S.; Meyer G. A.; Macedo J.; Amorim W.; Viana A. M. Repetitive somatic embryogenesis of Ocotea catharinensis Mez. (Lauraceae): effect of somatic embryo developmental stage and dehydration. Plant Cell, Tissue and Organ Culture 78: 55–62; 2004b.

    Google Scholar 

  • Siragusa M.; Carra A.; Salvia L.; Puglia A. M.; Pasquale F.; Carimi F. Genetic instability in calamondin (Citrus madurensis Lour.) plants derived from somatic embryogenesis induced by diphenylurea derivatives. Plant Cell Reports 26: 1289–1296; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Trebitsh T.; Staub J. E.; O’neill S. D. Identification of a 1-aminocyclopropane-1-carboxylic acid synthetase gene linked to the female (F) locus that enhances female sex expression in cucumber. Plant Physiology 113: 987–995; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Viana A. M.; Mantell S. H. Somatic embryogenesis of Ocotea catharinensis: an endangered tree of the Mata Atlântica (S. Brazil). In: Jain S. M.; Gupta P. K.; Newton R. J. (eds) Somatic Embryogenesis in Woody Plants, vol 5. Kluwer Academic Publishers, Dordrecht, pp 3–30; 1999.

    Google Scholar 

  • Vieira M. L. C.; Fungaro M. H. P.; Jubier M. F.; Lejeune B. Determination of taxonomic relationships among Brazilian taxa of Stylosanthes Sw., leguminosae, using RAPD markers. Pesquisa Agropecuária Brasileira 32: 305–310; 1997.

    Google Scholar 

  • Wang N. N.; Yang S. F.; Charng Y. Y. Differential expression of 1-aminocyclopropane-1-carboxylate synthase genes during orchid flower senescence induced by the protein phosphatase inhibitor okadaic acid. Plant Physiology 126: 253–260; 2001.

    Article  PubMed  CAS  Google Scholar 

  • White P. J. Recent advances in fruit development and ripening: an overview. Journal of Experimental Botany 53: 1995–2000; 1999.

    Article  Google Scholar 

  • Xiong L. Z.; Xu C. G.; Saghai Maroof M. A.; Zhang Q. F. Patterns of cytosine methylation in an elite rice hybrid and its parental lines by a methylation-sensitive amplification polymorphism technique. Molecular & General Genetics 261: 439–446; 1999.

    Article  CAS  Google Scholar 

  • Xu M.; Li X.; Korban S. S. AFLP-based detection of DNA methylation. Plant Molecular Biology Reporter 18: 361–368; 2000.

    Article  CAS  Google Scholar 

  • Xu M.; Li X.; Korban S. S. DNA-methylation alterations and exchanges during in vitro cellular differentiation in rose (Rosa hybrida L.). Theoretical and Applied Genetics 109: 899–910; 2004.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Brazilian institution ‘Fundação de Amparo à Pequisa do Estado de São Paulo, Grant no. 2004/03333-1, and to Christine Petrina Stock for the proofreading of the manuscript.

Author information

Authors and Affiliations

  1. Departamento de Genética, Escola Superior de Agricultura “Luiz de Queiroz”, Universidade de São Paulo, P.O. Box 83, 13400-970, Piracicaba, Brazil

    Luiz Ricardo Hanai & Maria Lucia Carneiro Vieira

  2. Instituto de Biociências, Universidade de São Paulo, 05422-970, São Paulo, Brazil

    Eny Iochevet Segal Floh

  3. Centro de Ciências Biológicas, Universidade Estadual de Londrina, 86051-990, Londrina, Brazil

    Maria Helena Pelegrinelli Fungaro & Francine Matias de Paula

  4. Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-602, Campos dos Goytacazes, Brazil

    Claudete Santa-Catarina

  5. Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, 88040-900, Florianópolis, Brazil

    Ana Maria Viana

Authors
  1. Luiz Ricardo Hanai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eny Iochevet Segal Floh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Helena Pelegrinelli Fungaro
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Claudete Santa-Catarina
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Francine Matias de Paula
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ana Maria Viana
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Maria Lucia Carneiro Vieira
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maria Lucia Carneiro Vieira.

Additional information

Editor: D. R. Duncan

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hanai, L.R., Floh, E.I.S., Fungaro, M.H.P. et al. Methylation patterns revealed by MSAP profiling in genetically stable somatic embryogenic cultures of Ocotea catharinensis (Lauraceae). In Vitro Cell.Dev.Biol.-Plant 46, 368–377 (2010). https://doi.org/10.1007/s11627-010-9291-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-010-9291-x

Keywords

Search

Navigation