Development of a common PVS2 vitrification method for cryopreservation of several fruit and vegetable crops

Many cryopreservation techniques are currently available, and it is common for new modifications to be developed for individual crops or specific genotypes. In this study, results of variations of the PVS2 cryopreservation protocol are compared to provide evidence for the suitability of a standard form of this technique for cryopreservation of a range of fruit, berry crops, and potato. Shoot cultures of Malus, Solanum, Lonicera, and Berberis were tested with variations of cold acclimation, pretreatment media, and PVS2 exposure times. A general protocol with some modifications was produced that was suitable for all four genera. The regenerative capacity of shoot tips after cryopreservation by this method exceeded a mean of 50% for Malus, Solanum, Lonicera, and Berberis, which is sufficient for setting storage in a cryobank. After liquid nitrogen storage, the shoot cultures that survived had a healthy appearance and developed rapidly. For each species tested, the only optimization required was the preparation of donor plants by cold acclimation and pretreatment. The choice of one common method simplifies the methodology for conducting experiments and storing a range of germplasm. The use of the PVS2 vitrification method with a 0.3-M sucrose pretreatment is multiuse and can be recommended as the most effective method for the cryopreservation of shoot tips from many plant species.