Cryopreservation of shoot apices and callus cultures of globe artichoke using vitrification method

Abstract

Background

Cryogenic cooling became a crucial tool for the storage of heterozygous plants such as globe artichoke. This study was carried out to optimize a reliable method for in vitro cryopreservation of shoot apices and callus cultures of globe artichoke using dimethylsulfoxide (DMSO) and Plant Vitrification Solutions 2 (PVS2) as cryoprotectant solutions. Shoot apices were exposed to DMSO or PVS2 for 20, 40, 60, and 80 min prior to plunge in liquid nitrogen (LN).

Results

It was found that using PVS2 as a cryoprotectant in cryopreservation of shoot apices of globe artichoke was more effective compared with using of DMSO alone. Among the exposure time tested, 60 min gave the best results of survival. The highest survival (60%), regeneration (56%), and proliferated shootlets (4.30) were obtained after cryoprotection with PVS2 for 60 min. Regarding callus cultures, the maximum values of fresh weights and subsequently growth value of recovered callus were registered with 40 min followed by 60 min exposure time. Related to the type of the tested cryoprotectants, the best survival and growth parameters of the cryopreserved callus cultures were obtained with PVS2 treatments. Treatment with PVS2 for 40 min registered the highest survival observations of cryopreserved callus. Also, the maximum values of fresh weight (1.30 g) and growth value (4.20) were obtained with 40 min exposure time. Microscopy analysis presented as cell morphology revealed that the treatment of PVS2 40% was the optimum for cell growth of cryopreserved callus of globe artichoke.

Conclusion

The results demonstrated that using PVS2 as a cryoprotectant in cryopreservation of shoot apices and callus cultures of globe artichoke was more effective compared with DMSO.