An improved cryopreservation protocol for pineapple apices using encapsulation-vitrification.
Several modifications to the cryogenic protocols previously described for pineapple apices were performed using vitrification and encapsulation-vitrification. Pregrowth of apices in sucrose-proline before loading significantly reduced the exposure duration to PVS2 and PVS3 required for successful cryopreservation. Encapsulation and treatments with PVS3 at 0 degree C gave the highest survival before and after cooling. Optimal conditions involved the encapsulation of pineapple apices in calcium alginate (3 percent) followed by a 2-d preculture in liquid medium with 0.16 M sucrose + 0.3 M proline for 24 h and then transfer to 0.3 M sucrose + 0.3 M proline for an additional 24 h. After preculture, samples were loaded in 0.75 M sucrose + 1 M glycerol solution at room temperature (25 min) and dehydrated with PVS3 at 0 degree C for 60 min before immersion into liquid nitrogen. Following this procedure 54 percent and 83 percent of apices from MD-2 and Puerto Rico varieties respectively survived.
Gámez-Pastrana
R
R
Universidad Veracruzana, Facultad de Ciencias Biológicas y Agropecuarias, Córdoba, Veracruz, México.
Martínez-Ocampo
Y
Y
Beristain
C I
CI
González-Arnao
M T
MT
D031230:Ananas; D015925:Cryopreservation / Q000379:methods*; D003681:Dehydration; D018520:Plant Shoots