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Development of a Cryopreservation Protocol for Vanilla Siamensis: An Endangered Orchid Species in Thailand
V V Chaipanich1, D L Roberts1, S Yenchon2
1Durrell Institute of Conservation and Ecology, School of Anthropology and Conservation, Faculty of Social Sciences, University of Kent, Kent, United Kingdom.
Background:
Vanilla siamensis is listed in Appendix-II of Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) as an endangered species in Thailand.
Objective:
To develop an optimum cryopreservation protocol for V. siamensis.
Materials And Methods:
Protocorms were precultured on solid ½ MS medium with 0.5 M sucrose for 0-7 d. For encapsulation-dehydration, encapsulated protocorms (beads) were dehydrated for 0-6 h. In the case of encapsulation-vitrification, the beads were loaded with a plant vitrification solution 2 (PVS2) at 0 ˚C for 0-90 min.
Results:
Protocorms precultured for 3 d gave the highest post-cryopreservation survival of 17%. Dehydration of the encapsulated protocorm beads for 4 h gave the highest survival of 33% and a regrowth of 25%. Protocorms subjected to the encapsulation-vitrification method did not survive at all.
Conclusion:
Protocorms precultured with 0.5 M sucrose for 3 d, encapsulated with 3% sodium alginate and dehydrated to a moisture content c. 14% before plunging into LN was a suitable method for the successful cryopreservation of V. siamensis protocorms.
