Cryopreservation of Arundina graminifolia (D. Don) Hochr. seeds using D cryo-plate method

In this study, cryopreservation of Arundina graminifolia seeds by D cryo-plate and encapsulation dehydration technique were investigated. The results showed that the D cryo-plate method gave the highest regrowth of 82%, followed by encapsulation-dehydration (74%). The D cryo-plate protocol is as follows: pour the alginate solution containing 2% (w/v) sodium alginate in calcium-free ½ MS basal medium with 0.4 M sucrose in the wells of the cryo-plate. For encapsulation-dehydration the protocol is the same as D cryo-plate without using cryo-plate. Place 20 to 50 seeds from 3-month-old selfing capsules in each well. Pour calcium chloride solution containing 0.1 M calcium chloride in ½ MS basal medium with 0.4 M sucrose. Expose to 0.4 M sucrose + 2 M glycerol (loading solution) for 30 min and then dehydrate under a laminar air-flow cabinet for 3 h. Put each cryo-plate in a 2-mL cryotube and plunge into liquid nitrogen for 1 d. Warm in 1.2 M sucrose solution (unloading solution) for 15 min then remove the solution and culture cryopreserved beads on ½ MS agar medium.