Cryopreservation of avocado embryogenic cultures using the droplet-vitrification method

This work reports on the cryopreservation of embryogenic cultures of avocado (Persea americana Mill.). Three cryopreservation protocols, based either on slow cooling or vitrification, were tested using two cell lines representative for the two types of embryogenic cultures that can be obtained in this species. Significant interactions between the embryogenic line and the cryopreservation protocol were observed. The best results were obtained when applying the droplet-vitrification method with recovery rates ranging from 77.78 to 100 %. The slow freezing method gave rise to different results depending on the cell line. While 80 % recovery was recorded in line D1, low recovery levels (33.33 %) were achieved in samples from line D31. The effect of different PVS2 incubation times was also evaluated and 60 min was considered as the optimum period. The developmental stage of starting material proved to be a key factor. Explant consisting of a mixture of embryogenic calli and somatic embryos at early stages resulted in the highest recovery rates after thawing.