Skip to main content
SpringerLink
Log in

Unfertilized ovary: a novel explant for coconut (Cocos nucifera L.) somatic embryogenesis

  • Cell Biology and Morphogenesis
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Unfertilized ovaries isolated from immature female flowers of coconut (Cocos nucifera L.) were tested as a source of explants for callogenesis and somatic embryogenesis. The correct developmental stage of ovary explants and suitable in vitro culture conditions for consistent callus production were identified. The concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) and activated charcoal was found to be critical for callogenesis. When cultured in a medium containing 100 μM 2,4-D and 0.1% activated charcoal, ovary explants gave rise to 41% callusing. Embryogenic calli were sub-cultured into somatic embryogenesis induction medium containing 5 μM abscisic acid, followed by plant regeneration medium (with 5 μM 6-benzylaminopurine). Many of the somatic embryos formed were complete with shoot and root poles and upon germination they gave rise to normal shoots. However, some abnormal developments were also observed. Flow cytometric analysis revealed that all the calli tested were diploid. Through histological studies, it was possible to study the sequence of the events that take place during somatic embryogenesis including orientation, polarization and elongation of the embryos.

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
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ABA:

Abscisic acid

2,4-D:

2,4-dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

NBB:

Naphthol blue black

PAS:

Periodic acid Schiff's reaction

CLZ:

Cambium-like zone

References

  • Bohanec B, Marijana J, Alojz I, Branka J (1995) Studies of gynogenesis in onion (Allium cepa L.) induction procedures and genetic analysis of regenerents. Plant Sci 104:215–224

    Article  CAS  Google Scholar 

  • Branton RL, Blake J (1983) Development of organized structures in callus derived from explants of (Cocos nucifera L.). Ann Bot 52:673–678

    Google Scholar 

  • Buffard-Morel J, Verdeil JL, Pannetier C (1988) Vegetative propagation of coconut palm through somatic embryogenesis, obtention of plantlets from leaf explant. In: Durand G, Bobichon L, Florent J (eds) Proceedings of the 8th international biotechnology symposium, Paris, p 117

  • Buffard-Morel J, Verdeil JL, Pannetier C (1992) Embryogenèse somatique du cocotier (Cocos nucifera L.) a partir d’explants foliaires: Etude histologique. Can J Bot 70:735–741

    Google Scholar 

  • Chan JL, Saenz L, Talavera C, Hornung R, Robert M, Oropeza C (1998) Regeneration of coconut (Cocos nucifera L.) from plumule explants through somatic embryogenesis. Plant Cell Rep 17:515–521

    Article  CAS  Google Scholar 

  • Compton ME (1994) Statistical methods suitable for the analysis of plant tissue culture data. Plant Cell Tissue Organ Cult 37:217–242

    Google Scholar 

  • Dolezel J, Binarova P, Lucretti S (1989) Analysis of nuclear DNA content in plant cells by flow cytometry. Biol Plant 31:113–120

    Google Scholar 

  • Ebert A, Taylor HF (1990) Assessment of the changes of 2,4-dichlorophenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal. Plant Cell Tissue Organ Cult 20:165–172

    CAS  Google Scholar 

  • Fernando SC, Gamage CKA (2000) Abscisic acid induced somatic embryogenesis in immature embryo explants of coconut (Cocos nucifera L.). Plant Sci 151:193–198

    Article  CAS  PubMed  Google Scholar 

  • Fernando SC, Verdeil JL, Hocher V, Weerakoon LK, Hirimburegama K (2003) Histological analysis of plant regeneration from plumule explants of Cocos nucifera L. Plant Cell Tissue Organ Cult 72:281–284

    Article  Google Scholar 

  • Fisher DB (1968) Protein staining of ribboned epon section for light microscopy. Histochemie 16:92–96

    Article  CAS  PubMed  Google Scholar 

  • George EF, Sherrington PD (1984) Plant propagation by tissue culture: handbook and directory of commercial laboratories. Exegenetics Eversley, London, pp 387–389

    Google Scholar 

  • Griffis JL, Litz RE (1997) Advances in the in vitro morphogenesis of several coconut (Cocos nucifera L.) tissues in Florida. In: Proceedings of the international cashew & coconut conference, Tanzania, pp 349–357

  • Gurel S, Gurel E, Kaya Z (2000) Double haploid plant production from unpollinated ovules of sugar beet (Beta vulgaris L). Plant Cell Rep 19:1155–1159

    Article  CAS  Google Scholar 

  • Hornung R (1995a) Micropropagation of (Cocos nucifera L.) from plumular tissues excised from mature zygotic embryos. Plant Rech Dev 212:38–41

    Google Scholar 

  • Hornung R (1995b) Initiation of callogenesis in coconut palm (Cocos nucifera L.). In: Oropeza C, Howard FW, Ashburner GR (eds) Lethal yellowing: research and practical aspects. Kluwer Academic, Dordrecht, pp 203–215

    Google Scholar 

  • Justin SHFW (1978) Vegetative propagation of coconuts. Report East Malling Research Station, 1977, pp 75–176

  • Karunaratne S, Periyapperuma K (1989) Culture of immature embryos of coconut (Cocos nucifera L.): callus proliferation and somatic embryogenesis. Plant Sci 62:247–253

    Article  Google Scholar 

  • Luthar Z, Bohanec B (1999) Induction of direct somatic embryogenesis in onion (Allium cepa L.) using a two step flower or ovary culture. Plant Cell Rep 18:797–802

    Article  CAS  Google Scholar 

  • Mukhambetzhanov SK (1997). Culture of non-fertilized female gametophytes in vitro. Plant Cell Tissue Organ Cult 48:111–119

    Article  Google Scholar 

  • Pannetier C, Buffard-Morel J (1982) Premiers résultats concernant la production d’embryons somatiques à partir de tissus foliaires de cocotier, Cocos nucifera L. Oleagineux 37:349–354

    Google Scholar 

  • Perez-Nunez MT, Chan JL, Saenz L, Gonzalez T, Verdeil JL, Oropeza C (2006). Improved somatic embryogenesis from coconut (Cocos nucifera L.) plumule explants cultured in vitro. In vitro Cell Dev Biol-Plant 42:37–43

    Article  Google Scholar 

  • Ruth SK, Stephen LS, John CB (1993) Ovule culture of sweet potato (Ipomoea batatas) and closely related species. Plant Cell Tissue Organ Cult 32:77–82

    Article  Google Scholar 

  • Sandoval A, Hocher V, Verdeil JL (2003) Flow cytometric analysis of the cell cycle in different coconut palm (Cocos nucifera L.) tissues cultured in vitro. Plant Cell Rep 22:25–31

    Article  CAS  PubMed  Google Scholar 

  • Schwendiman J, Pannetier C, Michaux-Ferriere N (1988) Histology of somatic embryogenesis from leaf explants of the palm Elaeis guineensis. Ann Bot 62:43–52

    Google Scholar 

  • Tang F, Tao Y, Zhao T, Wang G (2006). In vitro production of haploid and double haploid plants from pollinated ovaries of maize (zea mays). Plant Cell Tissue Organ Cult 84:233–237

    Article  Google Scholar 

  • Van Creij MGM, Kerckhoffs DMFJ, De Bruijn SM, Vreugdenhil D, Van Tuyl JM (2000) The effect of medium composition on ovary-slice culture and ovule culture in interspecific Tulipa gesneriana crosses. Plant Cell Tissue Organ Cult 60:61–67

    Article  Google Scholar 

  • Van Tuyl JM, Van Dien MP, Van Creij MGM, Van Kleinwee TCM, Franken J, Bino RJ (1991) Application of in vitro pollination, ovary culture, ovule culture and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses. Plant Sci 74(1):115–126

    Article  Google Scholar 

  • Verdeil JL, Buffard-Morel J (1995) Somatic embryogenesis in coconut (Cocos nucifera L.). In: Bajaj YPS (ed) Somatic embryogenesis and synthetic seed I. Biotechnology in agriculture and forestry, vol 30. Springer, Berlin Heidelberg New York, pp 299–317

  • Verdeil JL, Huet C, Grosdemange F, Buffard-Morel J (1994) Plant regeneration from cultured immature inflorescence of coconut (Cocos nucifera L.): evidence for somatic embryogenesis. Plant Cell Rep 13:218–221

    Article  CAS  Google Scholar 

  • Verdeil JL, Hocher V, Huet C, Grosdemange F, Escoute J, Ferriere N, Nicole M (2001) Ultra structural changes in coconut calli associated with the acquisition of embryogenic competence. Ann Bot 88:9–18

    Article  Google Scholar 

  • Weerakoon LK (2004) Coconut tissue and embryo culture in Sri Lanka: current developments and future challenges. In: Peiris TSG, Ranasinghe CS (eds) Proceedings of the international conference of the Coconut Research Institute of Sri Lanka – Part 1 (Review papers and guest presentations). CRI, Lunuwila, Sri Lanka pp 41–61

Download references

Acknowledgements

We gratefully acknowledge the financial assistance provided by the Institut de Recherche pour le Developpement (IRD), Montpellier, France and Coconut Research Institute, Sri Lanka. We are grateful to C. Duperray (INSERM, Montpellier, France) for the technical assistance in cytometry. We are thankful to Jacques Escoute (CIRAD, Monpellier, France) for the extended assistance to improve the quality of the pictures. We are also thankful to Dr. T. S. G. Peiris, Dr. S. Samita and J. D. J. S. Kularatne for their assistance in statistical analysis.

Author information

Authors and Affiliations

  1. Tissue Culture Division, Coconut Research Institute, 61150, Lunuwila, Sri Lanka

    Prasanthi I. P. Perera & L. Kaushalya Weerakoon

  2. Institute for Research and Development (IRD), UMR 1098 BEPC, IRD, BP 64501-911 Avenue Agropolis, 34394, Montpellier Cedex 1, France

    Valerie Hocher & Sylvie Doulbeau

  3. CIRAD, TA40/02 Avenue Agropolis, 34398, Montpellier Cedex 5, France

    Jean Luc Verdeil

  4. Department of Botany, Faculty of Science, University of Peradeniya, Peradeniya, Sri Lanka

    Deepthi M. D. Yakandawala

Authors
  1. Prasanthi I. P. Perera
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Valerie Hocher
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jean Luc Verdeil
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sylvie Doulbeau
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Deepthi M. D. Yakandawala
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L. Kaushalya Weerakoon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Prasanthi I. P. Perera.

Additional information

Communicated by P. Debergh

Rights and permissions

Reprints and permissions

About this article

Cite this article

Perera, P.I.P., Hocher, V., Verdeil, J.L. et al. Unfertilized ovary: a novel explant for coconut (Cocos nucifera L.) somatic embryogenesis. Plant Cell Rep 26, 21–28 (2007). https://doi.org/10.1007/s00299-006-0216-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-006-0216-4

Keywords

Search

Navigation