Efficient Method of Transporting Coconut (Cocos nucifera L.) Zygotic Embryos for Cryopreservation of Plumules by Encapsulation/Dehydration

  • H D D Bandupriya
Keywords: Coconut, cryopreservation, solidified agar medium, sucrose, ABA


Coconut is both socially and economically important crop in tropical and subtropical countries, thus the conservation of existing diversity of its germplasm is vital to maintain biodiversity, sustain crop production and utilisation of germplasm for crop improvement strategies. The recalcitrant storage behavior and large size of the coconut seed make it impossible to use as a germplasm storage material. Cryopreservation is an ideal means of long-term storage of germplasm which offers long-term storage capability with minimal storage space and maintenance requirements. The coconut embryo has been now adapted by various researchers for the purpose of germplasm exchange and it is now being routinely applied in germplasm collection and exchange activities with sufficient germination rates. The aim of the present study was to determine the effect of different coconut embryo transport/ storage methods [as solid endosperm plugs under cold temperature, embryos cultured in Solidified Agar Medium (SAM) or KCl solution under room temperature] on cryopreservation of plumules using encapsulation/dehydration method. The results revealed that plumules excised from embryos transported/ stored in SAM and pretreated with 1.0M sucrose could be cryopreserved with 71.8% survival and 56% recovery rates. The survival and recovery could be further increased up to 77.5% and 65% respectively by supplementation of 1.0M sucrose with 20 µM ABA.

Author Biography

H D D Bandupriya

Coconut Research Institute, Lunuwila, Sri Lanka


Assy-Bah, B., Durand-Gasselin, T. and Pannetier C. (1987) Use of zygotic embryo culture to collect germplasm of coconut (Cocos nucifera L.). FAO/IBPGR Plant Genetic Resource Newsletter 71: 4-10.
Assy-Bah, B. and Engelmann, F. (1992 a) Cryopreservation of immature embryos of coconut (Cocos nucifera L.). CryoLetters 13: 67-74.
Assy-Bah, B. and Engelmann, F. (1992 b) Cryopreservation of mature embryos of coconut (Cocos nucifera L.) and subsequent regeneration of plantlets. CryoLetters 13: 117-126.
Bajaj, Y.P.S. (1984) Induction of growth in frozen embryos of coconut and ovules of Citrus. Current Science 53:1215–1216.
Bandupriya, H.D.D., Fernando, S.C., Verdeil, J-L. and Malaurie, B. (2007) Effect of Abscisic acid on survival and recovery of cryopreserved plumule explants of coconut (Cocos nucifera L.). Cocos 18: 45-51.
Batugal, P. (2005) The International Coconut Genetic Resources Network (COGENT); its history and achievements. Coconut genetic resources (eds. P. Batugal, V. Ramanatha Rao, J. Oliver), pp 482–499. IPGRI-APO (Bioversity International), Serdang, Selangor, Malaysia.
Batugal, P. and Jayashree, K. (2005) COGENT’s multi-site International Coconut Genebank. Coconut genetic resources (eds. P. Batugal, V. Ramanatha Rao, J. Oliver), pp 106–114. IPGRI-APO (Bioversity International), Serdang, Selangor, Malaysia.
Chan, J.L., Saenz, L., Talavera, C., Hornung, R., Robert, M. and Oropeza, C. (1998) Regeneration of coconut (Cocos nucifera L.) from plumule explants through somatic embryogenesis. Plant Cell Reports 17:515–521.
Chin, H.F. and Pritchard, H.W. (1988) Recalcitrant seeds, a status report. International Board for Plant Genetic Resources, Rome.
Chin, H.F., Krishnapillay, B. and Hor, Y.L. (1989) A note on the cryopreservation of coconut embryos from young coconuts (Cocos nucifera L. var. Mawa). Pertanika 12 (2):183–186.
Diekmann, M. (1999) The use of biotechnology for the safe movement of coconut germplasm. Current Advances in Coconut Biotechnology (eds. C. Oropeza, J-L.Verdeil, G.R. Ashburner, R. Cardena and J.M. Santamaria), pp 259. Kluwer Academic Publishers, Dordrecht, Netherlands.
Engelmann, F. (2002) Coconut. In vitro collecting techniques for germplasm conservation (eds. V.C. Penc, J.A. Sandoval, V.M.A.Villalobos and F. Engelmann), pp 68–71. IPGRI technical bulletin No 7 IPGRI, Rome.
Engelmann, F. (1997) In vitro conservation methods. Biotechnology and plant genetic resources: Conservation and use (eds. B.V. Ford-Lloyd, H.J. Newbury and J.A. Callow), pp119-162. CABI, UK.
Fang, J.Y., Wetten, A. and Hadley, P. (2004) Cryopreservation of cocoa (Theobroma cocoa L.) somatic embryos for long term germplasm storage. Plant Science 166:669-675.
Ford, C.S., Jones, N.B. and van Staden, J. (2000) Cryopreservation and plant regeneration from somatic embryos of Pinus patula. Plant Cell Reports 19: 610–615.
Frison, E.A., Putter, C.A.J. and Diekmann, M. (1993) FAO/IBPGR technical guidelines for the safe movement of coconut germplasm. Food and Agricultural Organization of the United Nations/International Board for Plant Genetic Resources, Rome.
Gale, S., John, A. and Benson, E.E. (2007) Cryopreservation of Picea sitchensis (Sitka spruce) embryogenic suspensor masses. CryoLetters 28:225–239.
Gonzalez-Arnao, M.T. and Engelmann, F. (2006) Cryopreservation of plant germplasm using the encapsulation-dehydration technique: review and case study on sugarcane. CryoLetters 27(3):155–168.
Hocher, V., Verdeil, J-L. and Malaurie, B. (2004) Cocos nucifera, Coconut. Biotechnology of Fruit and Nut Crops (eds. R.E. Litz), pp 90-112. CABI Publishing Tropical Research and Education Center, University of Florida, USA.
Hornung, R., Domas, R. and Lynch, P.T. (2001) Cryopreservation of plumular explants of coconut (Cocos nucifera L.) to support programmes for mass clonal propagation through somatic embryogenesis. CryoLetters 22: 211-220.
Karun, A. (2001) Embryo culture protocol for collection and exchange of coconut germplasm. Indian Coconut Journal 31 (9): 17-18.
Karun, A., Sajini, K. K. S., Nair, M., Kumaran, P. M. and Samsudheen, K. (2006) Cryopreservation of coconut (Cocos nucifera L.) pollen. Journal of Plantation Crops 3: 568-571.
Karun, A. and Sajini, K. K. (2010) Cryopreservation of coconut zygotic embryos and pollen. Central Plantation Crops Research Institute, Kasaragod, India.
Kendall, E.J., Kartha, K.K., Qureshi, J.A. and Chermak, P. (1993) Cryopreservation of immature spring wheat zygotic embryos using an abscisic acid pretreatment. Plant Cell Reports 12 (2): 89-94.
Khoddamzadeh, A., Sinniah, U., Lynch, P., Abdul Kadir, M., Kadzimin, S. and Mahmood, M. (2011) Cryopreservation of protocorm-like bodies (PLBs) of Phalaenopsis bellina (Rchb.f.) christenson by encapsulation-dehydration. Plant Cell Tissue and Organ Culture 107 (3): 471-481.
Malaurie, B. (2001) Medium and long term conservation and safe international exchange of germplasm from food and cash tropical crops. Acta Horticulturae 560: 69-77.
Malaurie, B., Bandupriya, H.D.D., Fernando, S.C. and Verdeil, J-L. (2006) Optimisation
du procédé de cryoconservation de la plumule de cocotier. Les Actes du BRG 6: 449-468.
Malaurie, B., Borges, M. and N'Nan, O. (2002) Research of an optimal cryopreservation process using encapsulation/ osmoprotection/ dehydration and encapsulation/osmoprotection /vitrification techniques on caulinary meristems of coconut (Cocos nucifera L.). Burotrop Bulletin 20:16.
Moorhead, A. (2012) South Pacific coconut gene bank under threat. Available at http://www.nature.com/news/south-pacific-coconut-gene-bank-under-threat-1.11792, Accessed 10 January 2013.
Na, H-Y. and Kondo, K. (1996) Cryopreservation of tissue-cultured shoot primordia from shoot apices of cultured protocorms in Vanda pumila following ABA preculture and desiccation. Plant Science 118 (2): 195-201.
N’Nan, O., Borges, M., Konan, J-L.K., Hocher, V., Verdeil, J.-L., Tregear, J., N’guetta, A.S.P., Engelmann, F. and Malaurie, B. (2012) A simple protocol for cryopreservation of zygotic embryos of ten accessions of coconut (Cocos nucifera L.) In Vitro Cellular and Developmental Biology-Plant 48:160–166.
N’Nan, O., Hocher, V., Verdeil, J-L., Konan, J-L., Ballo, K., Mondeil, F. and Malaurie, B. (2008) Cryopreservation by encapsulation-dehydration of plumules of coconut (Cocos nucifera L.). CryoLetters 29 (4): 339-350.
Padro, M.D.A., Frattarelli, A., Sgueglia, A., Condello, E., Damiano, C. and Caboni, E. (2012) Cryopreservation of white mulberry (Morus alba L.) by encapsulation-dehydration and vitrification. Plant Cell Tissue and Organ Culture 108:167–172.
Paul, H., Daigny, G. and Sangwan-Noreel, B.S. (2000). Cryopreservation of apple (Malus x domestica Borkh.) shoot tips following encapsulation-dehydration or encapsulation-vitrification. Plant Cell Reports 19: 768-774.
Pence, V.C. (1998). Cryopreservation of bryophytes: The effects of abscisic acid and encapsulation dehydration. Bryologist 101: 278–281.
Rajasekaran, K. (1996) Regeneration of plants from cryopreserved embryogenic cell suspension and callus cultures of cotton (Gossypium hirsutum L.). Plant Cell Reports 15: 859-864.
Reed, B.M. (1990) Survival of in Vitro-grown apical meristems of Pyrus following Cryopreservation. HortScience 25: 111-113.
Rillo, E.P. and Paloma, M.B.F. (1991) Storage and transport of zygotic embryos of Cocos nucifera L. for in vitro culture. FAO/IBPGR Plant Genet Resources Newsletter 86:1-4.
Sakai, A. and Engelmann, F. (2007). Vitrification, encapsulation-vitrification and droplet-vitrification: a review. CryoLetters 28:151–172.
Senaratna, T., McKersie, B.D. and Bowley, S.R. (1989) Desiccation tolerance of alfalfa (Medicago sativa L.) somatic embryos. Influence of abscisic acid, stress pretreatments and drying rates. Plant Science 65: 253-259.
Sharaf, S.A., Shibli, R.A., Kasrawi, M.A. and Baghdadi, S.H. (2012) Cryopreservation of wild Shih (Artemisia herba-alba Asso.) shoot-tips by encapsulation-dehydration and encapsulation-vitrification. Plant Cell Tissue and Organ Culture 108: 437–444.
Shibli, R.A., Haagenson, D., Cunningham, S., Berg, W. and Volenec, J. (2001) Cryopreservation of alfalfa (Medicago sativa L.) cells by encapsulation-dehydration. Plant Cell Reports 20 (5): 445–450.
Sisunandar, Sopade, P.A., Samosir, Y.M.S., Rival, A. and Adkins, S.W. (2010) Dehydration improves cryopreservation of coconut (Cocos nucifera L.). Cryobiology 61:289–296.
Touchell, D.H., Chiang, V.L. and Tsai, C.J. (2002) Cryopreservation of embryogenic cultures of Picea mariana (black spruce) using vitrification. Plant Cell Reports 21: 118–124.
Uchendu, E.E., Brown, D.C.W. and Saxena, P.K. (2011) Cryopreservation of shoot tips and cotyledons of the North American ginseng (Panax Quinquefolius L.) CryoLetters 32(6): 463-472.
Wang, Q.C., Tanne, E., Arav, A. and Gafny, R. (2000) Cryopreservation of in vitro-grown shoot tips of grapevine by encapsulation–dehydration. Plant Cell Tissue and Organ Culture 63: 41–46.
Wang, Q.C., Batuman, O., Li, P., Bar-Joseph, M. and Gafny, R. (2002) Cryopreservation of in vitro grown shoot tips of ‘Troyer’ citrange [Poncirus trifoliata Raf.×Citrus sinensis (L.) Osbeck.] by encapsulation–dehydration. Plant Cell Reports 20: 901–906.
Werner, O., Espin, R.M.R., Bopp, M. and Atzorn, R. (1991). Abscisic acid induced drought tolerance in Funaria hygrometrica Hedw. Planta 186: 99–103.
How to Cite
H D D Bandupriya. (2014). Efficient Method of Transporting Coconut (Cocos nucifera L.) Zygotic Embryos for Cryopreservation of Plumules by Encapsulation/Dehydration . CORD, 30(2), 12. https://doi.org/10.37833/cord.v30i2.76