Skip to main content
SpringerLink
Log in

Somatic embryogenesis induction in leaves and petioles of a mature wild olive

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

We describe a protocol for somatic embryogenesis (SE) induction from an adult wild olive tree (Olea europaea ssp. europaea var. sylvestris. The protocol used confirms for the first time that there is no need to use juvenile or rejuvenated material for SE induction. For SE induction, petiole and leaf (proximal, intermediary and distal zones) explants were grown on Murashige and Skoog (MS) or Olive Medium (OM) media with different combinations of plant growth regulators (PGR): α- naphthaleneacetic acid (NAA), Zeatin (Zea), indole-3-butyric acid (IBA), 2-isopentyl adenine (2iP), thidiazuron (TDZ) and 6-benzylaminopurine (BAP). All media had 30 g/l sucrose and 7 g/l agar, and the pH was adjusted to 5.8. Cultures were incubated in the dark and, after 3 months, they were transferred to MS medium without PGR for expression. Petiole explants gave the highest callus production, while for SE induction and expression distal blade leaf and petiole explants gave the highest rates. The best medium for SE induction was MS with 12.25 μM IBA plus 4.56 μM Zea. Histological analyses confirmed the individuality of globular somatic embryos. This is the first report of SE expression in explants without rejuvenation in Olea genus, and opens perspectives for using this strategy in SE protocols both for this wild genotype and for commercial genotypes.

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

Fig. 1

Similar content being viewed by others

Abbreviations

MS:

Murashige and Skoog

OM:

Olive medium

BAP:

6-benzylaminopurine

IBA:

Indole-3-butyric acid

2iP:

2-Isopentyl adenine

NAA:

α- Naphthaleneacetic acid

SE:

Somatic embryogenesis

PGR:

Plant growth regulator

SEM:

Scanning electron microscopy

Zea:

Zeatin

References

  • Annarita (2009) Morphological evaluation of olive plants propagated in vitro culture through axillary buds and somatic embryogenesis methods. Afr J Plant Sci 3(3):037–043

    Google Scholar 

  • Benelli C, Fabbri A, Grassi S, Lambardi M, Rugini E (2001) Histology of somatic embryogenesis in mature tissues of olive (Olea europaea L.). J Hortic Sci Biotechnol 76:112–119

    Google Scholar 

  • Brito G, Costa A, Fonseca H, Santos C (2003) Response of Olea europaea subsp. maderensis shoots to osmotic stress induced by sorbi. Sci Hortic 97:411–417

    Article  CAS  Google Scholar 

  • Brito G, Loureiro J, Lopes T, Rodriguez E, Santos C (2007) Genetic characterization of olive trees from Madeira Archipelago using flow cytometry and microsatellite markers. Genet Resour Crop Evol 55:657–664. doi:10.1007/s10722-007-9268-8

    Article  Google Scholar 

  • Brito G, Costa A, Coelho C, Santos C (2009) Large-scale field acclimatization of Olea maderensis micropropagated plants: morphological and physiological survey. Trees 23:1019–1031

    Article  Google Scholar 

  • Capelo A, Brito G, Costa A, Santos C (2003) Micropropagation of the native Porto Santo Olea europaea ssp. maderensis: a contribution to combat desertification. I Simpósio sobre Biodiversidade em Ecossistemas Insulares. Book of Abstracts, Porto Santo

  • Chung HH, Chen JT, Chang WC (2007) Plant regeneration through direct somatic embryogenesis from leaf explants of Dendrobium. Biol plant 51(2):346–350

    Article  CAS  Google Scholar 

  • Conde P, Loureiro J, Santos C (2004) Somatic embryogenesis and plant regeneration from leaves of Ulmus minor Mill. Plant Cell Rep 22:632–639

    Article  CAS  PubMed  Google Scholar 

  • De Casas RR, Besnard G, Schönswetter P, Balaguer L, Vargas P (2006) Extensive gene flow blurs phylogeographic but not phylogenetic signal in Olea europaea L. Theor Appl Genet 113:575–583

    Article  Google Scholar 

  • Debnath CS (2009) A two-step procedure for adventitious shoot regeneration on excised leaves of lowbush blueberry. In Vitro Cell Dev Biol Plant 45:122–128

    Article  Google Scholar 

  • Fehér A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Review of plant biotechnology and applied genetics. Plant Cell Tiss Org Cult 74:201–228

    Article  Google Scholar 

  • Fournier D, Lejeune F, Tourte Y (1995) Cytological events during the inititation of meristematic nodules in callus derived from eggplant protoplasts. Biol Cell 85:93–100

    Article  CAS  PubMed  Google Scholar 

  • Leva A, Muleo R, Petruccelli R (1995) Long-term somatic embryogenesis from immature olive cotyledons. J Hortic Sci 70:417–421

    Google Scholar 

  • Lopes T, Pinto G, Loureiro J, Costa A, Santos C (2006) Determination of genetic stability in long-term somatic embryogenic cultures and derived plantlets of cork oak using microsatellite markers. Tree Physiol 26:1145–1152

    CAS  PubMed  Google Scholar 

  • Lopes T, Capelo A, Brito G, Santos C (2008) Genetic variability analyses of the somatic embryogenesis induction process in Olea spp. using nuclear microsatellites. Trees 23:29–36

    Article  Google Scholar 

  • Loureiro J, Capelo A, Brito G, Rodriguez E, Silva S, Pinto G, Santos C (2007) Micropropagation of Juniperus phoenicea L. adult plants and ploidy stability analyses in micropropagated plants using flow cytometry. Biol Plant 51(1):7–14

    Article  CAS  Google Scholar 

  • Merkle SA, Neu KA, Battle PJ, Bailey RL (1998) Somatic embryogenesis and plantlet regeneration from immature and mature tissues of sweetgum (Liquidambar styraciflua). Plant Sci 132:169–178

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–479

    Article  CAS  Google Scholar 

  • Pinto G, Valentim H, Costa A, Castro S, Santos C (2002a) Somatic embryogenesis in leaf callus from a mature Quercus suber L. tree. In Vitro Cell Dev Biol Plant 38:569–572

    Article  CAS  Google Scholar 

  • Pinto G, Santos CV, Neves L, Araújo C (2002b) Somatic embryogenesis and plant regeneration in Eucalyptus globulus. Plant Cell Rep 21:208–213

    Article  CAS  Google Scholar 

  • Rugini E (1984) In vitro-propagation of some olive (Olea europaea sativa L.) cultivars with different root-ability, and medium development using analytical data from developing shoots and embryos. Sci Hortic 24:123–134

    Article  CAS  Google Scholar 

  • Rugini E (1995) Somatic embryogenesis in olive (Olea europaea L.). In: Jain M, Gupta K, Newton J (eds) Somatic embryogenesis in woody plants. Kluwer, Dordrect, pp 171–189

    Google Scholar 

  • Rugini E, Caricato G (1995) Somatic embryogenesis and plant-recovery from mature tissues of olive cultivars (Olea europaea L.) “Canino” and “Moraiolo”. Plant Cell Rep 14:257–260

    Article  CAS  Google Scholar 

  • Santos CV, Brito G, Pinto G, Fonseca H (2003) In vitro plantlet regeneration of Olea europaea ssp. maderensis. Sci Hortic 97:83–87

    Article  CAS  Google Scholar 

  • Shibli RA, Shatnawi M, Abu E, Al-Juboory KH (2001) Somatic embryogenesis and plant recovery from callus of ‘Nabali’ olive (Olea europaea L.). Sci Hortic 88:243–256

    Article  Google Scholar 

  • Tang H, Ren Z, Reustle G, Krczal G (2002) Plant regeneration from leaves of sweet and sour cherry cultivars. Sci Hortic 93:235–244

    Article  CAS  Google Scholar 

  • Trabelsi EB, Bouzid S, Bouzid M, Elloumi N, Belfeleh Z, Benabdallah A, Ghezel R (2003) In vitro regeneration of olive tree by somatic embryogenesis. J Plant Biol 46(3):173–180

    Article  CAS  Google Scholar 

  • Trindade H, Pais MS (2003) Meristematic nodule culture: a new pathway for in vitro propagation of Eucalyptus globulus. Trees 17:308–315

    Google Scholar 

  • Williams EG, Maheswaran G (1986) Somatic embryogenesis—factors influencing coordinated behaviour of cells as an embryogenic group. Ann Bot 57(4):443–462

    Google Scholar 

Download references

Acknowledgments

Thanks are due to Dr Gloria Pinto and Eng. A. Costa

Author information

Authors and Affiliations

  1. University of Aveiro, Aveiro, Portugal

    Ana Margarida Capelo, Sónia Silva, Gina Brito & Conceição Santos

Authors
  1. Ana Margarida Capelo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sónia Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gina Brito
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Conceição Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ana Margarida Capelo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Capelo, A.M., Silva, S., Brito, G. et al. Somatic embryogenesis induction in leaves and petioles of a mature wild olive. Plant Cell Tiss Organ Cult 103, 237–242 (2010). https://doi.org/10.1007/s11240-010-9773-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-010-9773-x

Keywords

Search

Navigation