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Embryo maturity plays an important role for the successful cryopreservation of coconut (Cocos nucifera)

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.

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References

  1. 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

    PubMed  Google Scholar 

  2. Abdelnour-Esquivel A, Villalobos V, Engelmann F (1992) Cryopreservation of zygotic embryos of Coffea spp. CryoLetters 13:297–302

    Google Scholar 

  3. Assy-Bah B, Engelmann F (1992a) Cryopreservation of immature embryos of coconut (Cocos nucifera L.). CryoLetters 13:67–74

    Google Scholar 

  4. Assy-Bah B, Engelmann F (1992b) Cryopreservation of mature embryos of coconut (Cocos nucifera L.) and subsequent regeneration of plantlets. CryoLetters 13:117–126

    Google Scholar 

  5. Bajaj YPS (1984) Induction of growth in frozen embryos of coconut and ovules of citrus. Current Sci 53:1215–1216

    Google Scholar 

  6. 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)

    Article  Google Scholar 

  7. 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

  8. 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

    CAS  PubMed  Article  Google Scholar 

  9. Broschat TK, Donselman H (1986) Factors affecting storage and germination of Chysalidocarpus lutescens seeds. J Am Soc Hort Sci 111:872–877

    CAS  Google Scholar 

  10. 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

    Google Scholar 

  11. 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

    Article  Google Scholar 

  12. Chaudhury R, Malik SK (2004) Desiccation and freezing sensitivity during seed development in jackfruit. Seed Sci Tech 32:785–795

    Article  Google Scholar 

  13. 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

    Google Scholar 

  14. 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

    Google Scholar 

  15. 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

    Chapter  Google Scholar 

  16. 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

    Article  Google Scholar 

  17. 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

  18. 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

    Article  Google Scholar 

  19. 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

    Chapter  Google Scholar 

  20. 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

    PubMed  Google Scholar 

  21. 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

    CAS  Article  Google Scholar 

  22. 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

    Google Scholar 

  23. 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

    CAS  Article  Google Scholar 

  24. 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

    Article  Google Scholar 

  25. Rasband WS (2006) ImageJ [Online]. Available by US National Institutes of Health. Retrieved from http://rsb.info.nih.gov/ij/. Accessed 2 Nov 2006

  26. 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

  27. 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

    Article  Google Scholar 

  28. Sisunandar, Sopade PA, Samosir Y, Rival A, Adkins SW (2010a) Dehydration improves cryopreservation of coconut (Cocos nucifera L.). Cryobiology 61:289–296

    CAS  PubMed  Article  Google Scholar 

  29. 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

    CAS  PubMed  Article  Google Scholar 

  30. 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

    PubMed  Google Scholar 

  31. Steinmacher DA, Saldanha CW, Clement CR, Guerra MP (2007) Cryopreservation of peach palm zygotic embryos. CryoLetters 28:13–22

    PubMed  Google Scholar 

  32. Thampan PK (1981) Handbook on coconut palm. Oxford and IBH Publishing Co, New Delhi, 302 pp

    Google Scholar 

  33. 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

    PubMed  Google Scholar 

  34. Wen B, Song S (2007) Acquisition of cryotolerance in maize embryos during seed development. CryoLetters 28:109–118

    PubMed  Google Scholar 

Download references

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.

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Correspondence to Steve W. Adkins.

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Editor: John Forster

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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

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Keywords

  • Embryo maturity
  • Cryopreservation
  • Coconut
  • Cocos nucifera