Three vitrification-based cryopreservation procedures cause different cryo-injuries to potato shoot tips while all maintain genetic integrity in regenerants

• Three vitrification-based cryogenic methods resulted in different patterns of cell injury.
• The injury was mainly caused by dehydration with PVS2 rather than freezing in LN.
• No genetic alterations were detected in all shoots regenerated from the three vitrification-based cryogenic methods by ISSR and AFLP.

We previously reported successful cryopreservation of shoot tips of potato ‘Zihuabai’ by three vitrification-based protocols. In the present study, cryo-injury to shoot tips and genetic stability in regenerants recovered from cryopreserved shoot tips by the three vitrification-based protocols were further investigated. The results showed that sucrose preculture caused no obviously different injuries, while dehydration with plant vitrification solution 2 (PVS2) was the step causing major damage to cells of shoot tips, regardless of the cryogenic procedures. Compared with droplet-vitrification and encapsulation–vitrification, vitrification caused the most severe injury to cells of the shoot tips, thus resulting in much longer time duration for shoot recovery and much lower shoot regrowth rate. Cells in apical dome and the youngest leaf primordia were able to survive and subsequently some of them regrew into shoots following all three vitrification-based cryopreservation procedures. Analyses using inter-simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP) markers in shoots regrown from all three vitrification-based protocols did not find any polymorphic bands. The results reported here suggest that vitrification-based cryo-procedures can be considered promising methods for long-term preservation of potato genetic resources.