Abstract
Genetic diversity of coconut (Cocos nucifera L.) is being lost due to a combination of pest and disease attack, urban encroachment, natural disasters, as well as introgression with exotic genetic types. Consequently, there is a need to undertake germplasm conservation before further loss occurs. Since coconut has a large, recalcitrant seed (sensitive to desiccation), it cannot be stored in traditional ways in a seed bank. Cryopreservation of zygotic embryos is now seen as an important storage approach although published techniques are still not reliable. Given the importance of embryo maturity to the success of cryopreservation in other species, the effect of coconut embryo maturity on cryopreservation success was investigated using four cultivars (‘Nias Yellow Dwarf’, ‘Tebing Tinggi Dwarf’, ‘Takome Tall’, and ‘Bali Tall’). After cryopreservation, using a new four-step protocol (rapid desiccation, rapid freezing, rapid thawing, and recovery and acclimatization for 4 mo in the glasshouse), we found that the embryos isolated from an 11-mo-old fruit gave the highest number of normal seedlings (~28%) when compared to counterparts excised from younger fruits. In addition, the results showed that fruit could be stored for up to 3 wk prior to embryo isolation before their performance in cryopreservation was compromised.
Similar content being viewed by others
References
Abdelnour-Esquivel A, Engelmann F (2002) Cryopreservation of chayote (Sechium edule Jacq. SW) zygotic embryos and shoot tips from in vitro plantlets. CryoLetters 23:299–308
Abdelnour-Esquivel A, Villalobos V, Engelmann F (1992) Cryopreservation of zygotic embryos of Coffea spp. CryoLetters 13:297–302
Assy-Bah B, Engelmann F (1992a) Cryopreservation of immature embryos of coconut (Cocos nucifera L.). CryoLetters 13:67–74
Assy-Bah B, Engelmann F (1992b) Cryopreservation of mature embryos of coconut (Cocos nucifera L.) and subsequent regeneration of plantlets. CryoLetters 13:117–126
Bajaj YPS (1984) Induction of growth in frozen embryos of coconut and ovules of citrus. Current Sci 53:1215–1216
Barbedo C, Barbedo A, Nakagawa J, Sato O (1999) The effect of fruit age and post-harvest period of cucumber on stored seeds. Pesq Agrop Brasileira 34:839–847 (in Portuguese)
Batugal P, Jayashree J (2005) COGENT's multi-site international coconut genebank. In: Batugal P, Ramanatha Rao V, Oliver J (eds) Coconut genetic resources. International Plant Genetic Resources Institute-Regional Office for Asia, the Pacific and Oceania (IPGRI-APO), Serdang, Selangor DE, Malaysia, pp 106–114
Berjak P, Pammenter NW, Vertucci CW (1992) Homiohydrous (recalcitrant) seed: developmental status, desiccation sensitivity and state of water in axes of Landolphia kirkii Dyer. Planta 186:249–261
Broschat TK, Donselman H (1986) Factors affecting storage and germination of Chysalidocarpus lutescens seeds. J Am Soc Hort Sci 111:872–877
Broschat TK, Donselman H (1987) Effect of maturity, storage, presoaking and seed cleaning on germination in three species of palms. J Environ Hort 5:6–9
Chandel KPS, Chudhury R, Radhamani J, Malik SK (1995) Desiccation and freezing sensitivity in recalcitrant seeds of tea, cocoa and jackfruit. Ann Bot 76:443–450
Chaudhury R, Malik SK (2004) Desiccation and freezing sensitivity during seed development in jackfruit. Seed Sci Tech 32:785–795
Chin HF, Krishnapillay B, Hor YL (1989) A note on the cryopreservation of embryos from young coconut (Cocos nucifera var MAWA). Pertanika 12:183–186
Demir I, Okcu G (2005) Effect of post-harvest maturation treatment on germination and potential longevity of pepper (Capsicum annuum) seed. Indian J Agric Sci 75:19–22
Engelmann F (1999) Cryopreservation of coconut germplasm. In: Oropeza C, Verdeil JL, Ashburner GR, Cardena R, Santamaria JM (eds) Current advances in coconut biotechnology. Kluwer Academic Publishers, Dordrecht, pp 289–296
Farant JM, Walters C (1998) Ultra structural and biophysical changes in developing embryos of Aesculum hippocatanum in relation to acquisition of tolerance to drying. Physiol Plant 104:513–524
Foale M (2003) The coconut odyssey: the bounteous possibilities of the tree of life. ACIAR Monograph No. 101, Australian Centre for International Agricultural Research, Canberra, 132 pp
Hong TD, Ellis RH (1990) A comparison of maturation drying, germination and desiccation tolerance between developing seeds of Acer pseudoplatanus L. and Acer platanoides L. New Phytol 116:589–596
Kenmode AR, Finch-Savage WE (2002) Desiccation sensitivity in orthodox and recalcitrant seed in relation to development. In: Black MJ, Pritchard HW (eds) Desiccation and survival in plants: drying without dying. CABI Publishing, Wallingford, pp 149–184
Kim HH, Cha YS, Baek HJ, Cho EG, Chae TA, Engelmann F (2002) Cryopreservation of tea (Camellia sinensis L.) seeds and embryonic axes. CryoLetters 23:209–216
Lehner A, Bailey C, Flechel B, Poels P, Come D, Corbineau F (2006) Changes in wheat seed germination ability, soluble carbohydrate and antioxidant enzyme activities in the embryo during the desiccation phase of maturation. J Cereal Sci 43:175–182
Nadarajan J, Staines HJ, Benson EE, Marzalina M, Krishnapillay B, Harding K (2007) Optimization of cryopreservation for Sterculia cordata zygotic embryos using vitrification technique. J Trop Forest Sci 19:79–85
Perera PIP, Hocher V, Verdeil JL, Bandupriya HDD, Yakandawala DMD, Weerakoon LK (2008) Androgenic potential in coconut (Cocos nucifera L.). Plant Cell Tiss Organ Cult 92:293–302
Pritchard HW, Prendergast G (1986) Effect of desiccation and cryopreservation on the in vitro viability of embryos of the recalcitrant seed species Araucaria hunsteinii K. Schum J Ex Bot 37:1388–1397
Rasband WS (2006) ImageJ [Online]. Available by US National Institutes of Health. Retrieved from http://rsb.info.nih.gov/ij/. Accessed 2 Nov 2006
Rillo EP (2004) Importing and growing embryos for the coconut genebank. In: Ikin R, Batugal P (eds) Germplasm health management for COGENT’s multi-site international coconut genebank. International Plant Genetic Resources Institute-Regional Office for Asia, the Pacific and Oceania (IPGRI-APO), Serdang, Selangor DE, Malaysia, pp 62–68
Samarah N, Allataifeh N, Turk M, Tawaha AR (2003) Effect of maturity stage on germination and dormancy of fresh and air-dried seeds of bitter vetch (Vicia ervilia L.). NZ J Agric Res 46:347–354
Sisunandar, Sopade PA, Samosir Y, Rival A, Adkins SW (2010a) Dehydration improves cryopreservation of coconut (Cocos nucifera L.). Cryobiology 61:289–296
Sisunandar RA, Turquay P, Samosir Y, Adkins SW (2010b) Cryopreservation of coconut (Cocos nucifera L.) zygotic embryos does not induce morphological, cytological or molecular changes in recovered seedlings. Planta 232:435–447
Sopade PA, Sopade PA, Samosir Y, Rival A, Adkins SW (2012) Conservation of coconut (Cocos nucifera L.) germplasm at sub-zero temperature. CryoLetters 33:465–475
Steinmacher DA, Saldanha CW, Clement CR, Guerra MP (2007) Cryopreservation of peach palm zygotic embryos. CryoLetters 28:13–22
Thampan PK (1981) Handbook on coconut palm. Oxford and IBH Publishing Co, New Delhi, 302 pp
Walters C, Touchell DH, Power P, Wesley-Smith J, Antolin MF (2002) A cryopreservation protocol for embryos of the endangered species Zizania texana. CryoLetters 23:291–298
Wen B, Song S (2007) Acquisition of cryotolerance in maize embryos during seed development. CryoLetters 28:109–118
Acknowledgments
The project was partly funded by the Australian Agency for International Development (AusAID), the Endeavour Research Fellowships Australia 2010, and the Australian Centre for International Agricultural Research, via project HORT/1998/061.
Author information
Authors and Affiliations
Corresponding author
Additional information
Editor: John Forster
Rights and permissions
About this article
Cite this article
Sisunandar, Novarianto, H., Mashud, N. et al. Embryo maturity plays an important role for the successful cryopreservation of coconut (Cocos nucifera). In Vitro Cell.Dev.Biol.-Plant 50, 688–695 (2014). https://doi.org/10.1007/s11627-014-9633-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-014-9633-1