Cryopreservation of Pineapple Shoot Tips by the Droplet Vitrification Technique

Abstract

Cryopreservation is a technique that allows the conservation of many species for long periods. Among the protocols used for cryopreservation, droplet vitrification has shown efficient results in preserving shoot tips of various wild and cultivated pineapple genotypes. The method consists of extraction of shoot tips from plants grown in vitro, dehydration for a period of 48 h in a preculture medium supplemented with a high concentration of sucrose, treatment in a plant vitrification solution (PVS2), and immersion in liquid nitrogen. The method described in this chapter has produced survival and regeneration indices of around 70%, depending on the genotype and physiological conditions of the initial explants. The objective of this chapter is to describe in detail a droplet vitrification protocol for shoot tips that is easy to perform for cryopreservation of pineapple germplasm.

Morpho-Anatomical and Physiological Assessments of Cryo-Derived Pineapple Plants (Ananas comosus var. comosus) after Acclimatization

“…Cryopreservation methods have already been developed for pineapple shoot tips [5,[7][8][9][10][11][12][13]. The vitrification-based cryopreservation method is the most widely applied for cryopreserving pineapple shoot tips [5,[10][11][12][13].…”

“…Cryopreservation methods have already been developed for pineapple shoot tips [5,[7][8][9][10][11][12][13]. The vitrification-based cryopreservation method is the most widely applied for cryopreserving pineapple shoot tips [5,[10][11][12][13]. In this method, precultured shoot tips are exposed to a highly concentrated plant vitrification solution (PVS) before being directly immersed in liquid nitrogen (LN) [2,12,13].…”

“…Droplet-vitrification uses ultra-fast cooling and warming rates of shoot tips, an important requirement for successful cryopreservation protocols [14,15]. Souza et al [10,11] described a successful droplet-vitrification protocol that was applied to 16 genotypes of Ananas, both wild and cultivated, belonging to four botanical varieties. In this procedure, shoot tips were precultured for 48 h in preculture medium containing 0.3 M sucrose and then transferred to aluminum foil in 4-5 µL of plant vitrification solution 2 (PVS2) and treated at 0 • C for 45 min before being directly submerged in LN.…”