Throughout the cryopreservation process, plants were exposed to a series of
abiotic stresses such as desiccation and osmotic pressure due to highly concentrated
vitrification solution. Abiotic stress stimulates the production of reactive oxygen species
(ROS) which include hydrogen peroxide, superoxide radicals, and singlet oxygen. Higher
production of ROS may lead to oxidative stress which contributes to the major injuries in
cryopreserved explants. Antioxidant enzymes in plant such as ascorbate peroxidase
(APX) can protect plants from cell damage by scavenging the free radicals. This study was
determined based on APX enzyme activity of Aranda Broga Blue orchid’s protocorm-like
bodies (PLBs) in response to PVS2 (Plant Vitrification Solution 2) cryopreservation
treatments at different stages. PLBs that were precultured at 0.25 M sucrose for 3 days
were subjected to vitrification cryopreservation method. Results obtained showed that the
highest APX activity was achieved at PVS2 cryoprotectant treatment prior liquid nitrogen
(LN) storage. This phenomenon indicating that accumulation of osmotic and dehydrating
stress throughout the cryopreservation treatment resulted in oxidative burst which in turn
leads to higher APX activity in order to control the excess production of ROS. To
conclude, PVS2 treatment was revealed as the most detrimental step throughout
cryopreservation treatment. Thus, this research also suggested that exogenous
antioxidant such as ascorbic acid can be added throughout cryopreservation procedure
especially at PVS2 treatment in the future experiments to aid in regrowth of cryopreserved
explants by reducing oxidative stress.