Embryo Incision as a New Technique for Double Seedling Production of Indonesian Elite Coconut Type “Kopyor”


  • Sisunandar Sisunandar Biology Education Department, University of Muhammadiyah Purwokerto Jl. Raya Dukuhwaluh, Kembaran, Purwokerto, 53182, Indonesia
  • Alkhikmah Alkhikmah Biology Education Department, University of Muhammadiyah Purwokerto Jl. Raya Dukuhwaluh, Kembaran, Purwokerto, 53182, Indonesia
  • Arief Husin Biology Education Department, University of Muhammadiyah Purwokerto Jl. Raya Dukuhwaluh, Kembaran, Purwokerto, 53182, Indonesia
  • Aman Suyadi Agrotechnology Department, Faculty of Agriculture, University of Muhammadiyah Purwokerto, Jl. Raya Dukuhwaluh, Kembaran, Purwokerto, 53182, Indonesia




embryo culture, embryo splitting, in vitro culture, meristem, multiplication


One of the present major limitations of seedling production of kopyor type coconut using embryo culture is that only one seedling can be produced from a single embryo. Therefore, we report on the development of a new breakthrough technique for the production of double seedlings from a single embryo. The technique consists of four steps, viz. (i) germination; (ii) incision; (iii) splitting; and (iv) recovery. A histological study was carried out on the development of the halved embryo into a new shoot. The best recovery process was obtained when the incised embryo was split into two and recovered into Murashige and Skoog (MS) medium supplemented with 2 μM IBA and 15 μM kinetin. Following this protocol, an average of 56 shoots was successfully recovered from 30 zygotic embryos. The histological study also revealed that the meristem tissue of the halved embryo was able to produce a new meristem and primordial leaf. Most of the shoots then went on to produce normal seedlings and could be acclimatized successfully after having developed 2 or 3 leaves. This protocol is useful for routine seedling production of the kopyor-type coconut.


FAO, FAOSTAT Data, http://faostat.fao.org/ (3 March 2011).

Mahmud, Z. & Ferry, Y., Prospects of Byproduct Processing of Coconut Perspektif, 4, pp. 55-63, 2005. (Text in Indonesian)

Maskromo, I., Mashud, N. & Novarianto, H., Potential Development of Kopyor Coconut in Indonesia, Warta Penelitian dan Pengembangan, 13, pp. 4-6, 2007. (Text in Indonesian)

Samosir, Y., Mashud, N., Novarianto, H., Lien, V.T.M., Rillo, E.P., Magdalita, P.M., Damasco, O.P., Kembu, A., Faure, M.G. & Adkins, S.W., A New Embryo Culture Protocol for Coconut Germplasm Conservation and Elite-Type Seedling Production, Australian Centre for International Agricultural Research, Canberra, 2008.

Sukendah, Sudarsono, Witjaksono & Khumaida, N., Developing Embryo Culture Technique for Kopyor Coconut (Cocos nucifera L.) Originated from Sumenep, East Java, through the Addition of Additive Substances and Testing Subculture Periods, Buletin Agronomi, 36, pp. 16-23, 2008. (Text in Indonesian)

Perez-Nunez, M.T., Chan, J.L., Saenz, T., Gonzales, T., Verdeil, J.L. & Oropeza, C., Improved Somatic Embryogenesis from Cocos nucifera L. Plumule Explans, In Vitro Plant Cellular & Developmental Biology, 42, pp. 37-43, 2006.

Perera, P.I.P., Hocher, V., Verdeil, J.L., Doulbeau, S., Yakandawala, D.M.D. & Weerakoon L.K., Unfertilized Ovary: A New Novel Explant for Coconut (Cocos nucifera L.) Somatic Embryogenesis, Plant Cell Report, 26, pp. 21-28, 2007.

Mashud, N., Developing More Efficient (30%) Embryo Culture Method for Kopyor Coconut, Laporan Penelitian Program Insentif Riset Terapan, Balai Penelitian Tanaman Kelapa dan Palma Lain, Manado, 2010. (Text In Indonesian)

Sukendah, In Vitro Multiplication and Molecular Analisys of Kopyor Coconut, Ph.D Dissertation, Postgraduate School Institut Pertanian Bogor, Bogor, 2009.

Nunez, T.C., Doubling Macapuno Seedling Production thru Embryo Splitting, Philippine Journal of Crop Science, 22(Suppl. No. 1), pp. 64, 1997.

Sisunandar, In Vitro Propagation Method of Kopyor Coconut Through Embryo Incision Technique, Patent Pending No. P00201201185, 2012.

Sisunandar, Rival, A., Turquay, P., Samosir, Y. & Adkins, S.W., Cryopreservation of Coconut (Cocos nucifera L.) Zygotic Embryos does not Induce Morphological, Cytological or Molecular Changes in Recovered Seedlings, Planta, 232, pp. 435-447, 2010.

Rillo, E.P., Importing and Growing Embryos for the Coconut Genebank, Germplasm Health Management for COGENT's Multi-site International Coconut Genebank, Ikin, R. & Batugal, P. (eds), International Plant Genetic Resources Institute-Regional Office for Asia, the Pacific and Oceania (IPGRI-APO), Serdang, Selangor DE, Malaysia, pp. 62-68, 2004.

Murashige, T. & Skoog, F., A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Culture, Physiologia Plantarum, 15, pp. 473-497, 1962.

Sass, J.E., Bitanical Microtechnique, The Iowa State College Press, Iowa, pp. 69-71, 1951.

Sisunandar, Sopade, P.A., Samosir, Y., Rival, A. & Adkins, S.W., Dehydration Improves Cryopreservation of Coconut (Cocos nucifera L.), Cryobiology, 61, pp. 289-296, 2010.

Sisunandar, Sopade, P.A., Samosir, Y., Rival, A. & Adkins, S.W., Conservation of Coconut (Cocos nucifera L.) Germplasm at Sub-zero Temperature, CryoLetters, 33, pp. 465-475, 2012.

George, E.F. & de Klerk, G.J., The Components of Plant Tissue Culture Media I: Macro- and Micro-nutrients, Plant Propagation by Tissue Culture 3rd Edition, George, E.F., Hall, M.A., Jan De Klerk, G. (eds), Springer, Dordrecht, The Netherlands, pp. 65-114, 2008.