Date Palm Biotechnology Protocols Volume II

Shawky A. Bekheet

In vitro technology offers a potential solution for the conservation of date palm germplasm. Slow growth induced by low temperature allows storage from several months up to few years. Otherwise, cryopreservation is suitable for long-term in vitro conservation, at between -79 and -196 °C. This chapter describes a protocol for cold storage at 5 °C and cryopreservation of date palm tissue cultures. For cold storage, 70% of shoot buds remain healthy after storing for 12 months at 5 °C, and callus cultures remain fully viable after 12 months of storage. For cryopreservation of embryogenic cultures using dehydration by air, apparently, 20 min air drying is the best for cryopreservation. Among different types of sugars used as osmotic agents in pre-culture medium, 1 M sucrose is the best for the survival of cryopreserved cultures. However, exposure of embryogenic cultures to vitrification solution for 60 min at 0 °C gives the highest percentage of survival and conversion to plantlets.