An improved cryopreservation protocol for pineapple apices using encapsulation-vitrification.

R Gámez-Pastrana; Y Martínez-Ocampo; C I Beristain; M T González-Arnao

PubMed · 15660167

An improved cryopreservation protocol for pineapple apices using encapsulation-vitrification.

R Gámez-Pastrana·Y Martínez-Ocampo·C I Beristain·M T González-Arnao

Abstract

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.

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R Gámez-Pastrana, Y Martínez-Ocampo, C I Beristain, M T González-Arnao. An improved cryopreservation protocol for pineapple apices using encapsulation-vitrification.. https://pubmed.ncbi.nlm.nih.gov/15660167/

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