Conservation of coconut (Cocos nucifera L.) germplasm at sub-zero temperature

Protocols are proposed for the low (-20 degree C) and ultra-low (-80 degree C) temperature storage of coconut (Cocos nucifera L.) embryos. A tissue dehydration step prior to storage, and a rapid warming step upon recovery optimized the protocol. The thermal properties of water located within embryos were monitored using differential scanning calorimetry (DSC). In the most efficient version of the protocol, embryos were dehydrated under a sterile air flow in a dehydration solution containing glucose (3.33 M) and glycerol (15 percent) for 16 hours. This protocol decreased the embryo water content from 77 to 29 percent FW and at the same time reduced the amount of freezable water down to 0.03 percent. The dehydrated embryos could be stored for up to 3 weeks at -20 degree C (12 percent producing normal plants upon recovery) or 26 weeks at -80 degree C (28 percent producing normal plants upon recovery). These results indicate that it is possible to store coconut germplasm on a medium term basis using an ultra-deep freezer unit. However for more efficient, long term storage, cryopreservation remains the preferred option.