Cryopreservation of embryogenic cell suspensions by encapsulation-vitrification

Area of Science:

• Plant Biotechnology
• Cryopreservation
• Germplasm Preservation

Background:

• Cryopreservation is crucial for conserving plant genetic resources.
• Traditional methods face limitations in long-term storage efficacy.
• Encapsulation-vitrification (EV) is an emerging technique combining encapsulation-dehydration and vitrification.

Purpose of the Study:

• To detail the encapsulation-vitrification protocol for plant germplasm cryopreservation.
• To establish a reliable method for grapevine (Vitis) cell suspension cryopreservation.
• To demonstrate the successful regeneration of whole plants from cryopreserved cells.

Main Methods:

• Grapevine cell suspensions were encapsulated in alginate beads.
• Beads underwent stepwise preculturing in increasing sucrose concentrations.
• Dehydration with PVS2 and rapid immersion in liquid nitrogen followed.

Main Results:

• Cryopreserved cells showed survival after rapid thawing and post-culture treatment.
• Surviving cells proliferated and re-established embryogenic cell suspensions.
• Regenerated whole plantlets were successfully developed from torpedo-stage embryos.

Conclusions:

• Encapsulation-vitrification is an effective cryopreservation method for grapevine embryogenic cell suspensions.
• The protocol facilitates long-term storage and subsequent regeneration of plant material.
• This technique holds significant potential for germplasm conservation in various plant species.