2015Acta Scientiarum AgronomyOpen access

Cryopreservation of the mangaba tree (Hancornia speciosa Gomes): a protocol for long-term storage

Paulo Augusto Almeida Santos, Renato Paiva, Luciano Coutinho Silva, Ana Cristina de Souza, Marlúcia Cruz de Santana, Diogo Pedrosa Corrêa da Silva

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Abstract

The aim of this study was to evaluate the efficiency of vitrification and droplet vitrification for the cryopreservation of Hancornia speciosa shoot tips. The shoot tips were subjected to four different periods of exposure (15, 30, 45 and 60 min.) to plant vitrification solution 2 (PVS2) before plunging into liquid nitrogen. We evaluated the regrowth of H. speciosa shoot tips that were cryopreserved by the classical vitrification technique and by the droplet vitrification technique. Shoot tips were submitted to different periods of pre-culture (absence, 24 or 48h) in a medium containing 0.3 M sucrose prior to cryopreservation. With a PVS2 exposure period of 60 min., significant differences were observed between the two techniques used in this study with respect to regrowth rates; however, both cryopreservation methods allowed for a shoot tip regrowth rate of over 70%. Use of a preculture step increased the regrowth rate of cryopreserved shoot tips. Shoot tip cryopreservation techniques employing vitrification and droplet vitrification proved to be viable for the long-term storage of mangaba tree samples.

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The aim of this study was to evaluate the efficiency of vitrification and droplet vitrification for the cryopreservation of Hancornia speciosa shoot tips. The shoot tips were subjected to four different periods of exposure (15, 30, 45 and 60 min.) to plant vitrification solution 2 (PVS2) before plunging into liquid nitrogen. We evaluated the regrowth of H. speciosa shoot tips that were cryopreserved by the classical vitrification technique and by the droplet vitrification technique. Shoot tips were submitted to different periods of pre-culture (absence, 24 or 48h) in a medium containing 0.3 M sucrose prior to cryopreservation. With a PVS2 exposure period of 60 min., significant differences were observed between the two techniques used in this study with respect to regrowth rates; however, both cryopreservation methods allowed for a shoot tip regrowth rate of over 70%. Use of a preculture step increased the regrowth rate of cryopreserved shoot tips. Shoot tip cryopreservation techniques employing vitrification and droplet vitrification proved to be viable for the long-term storage of mangaba tree samples.

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Available abstract

The aim of this study was to evaluate the efficiency of vitrification and droplet vitrification for the cryopreservation of Hancornia speciosa shoot tips. The shoot tips were subjected to four different periods of exposure (15, 30, 45 and 60 min.) to plant vitrification solution 2 (PVS2) before plunging into liquid nitrogen. We evaluated the regrowth of H. speciosa shoot tips that were cryopreserved by the classical vitrification technique and by the droplet vitrification technique. Shoot tips were submitted to different periods of pre-culture (absence, 24 or 48h) in a medium containing 0.3 M sucrose prior to cryopreservation. With a PVS2 exposure period of 60 min., significant differences were observed between the two techniques used in this study with respect to regrowth rates; however, both cryopreservation methods allowed for a shoot tip regrowth rate of over 70%. Use of a preculture step increased the regrowth rate of cryopreserved shoot tips. Shoot tip cryopreservation techniques employing vitrification and droplet vitrification proved to be viable for the long-term storage of mangaba tree samples.

Key concepts: Cryopreservation, Biology, Cell biology, Embryo

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