Abstract
This study aimed to examine the effects of ice crystals and oxidative stress on seed viability in the context of cryopreservation, with the ultimate goal of identifying potential solutions to address the persistently low regeneration rate observed in recalcitrant medicinal plant seeds following cryopreservation. Using differential scanning calorimetry technology alongside seeds germination at different moisture levels after cryopreservation helped determine the best moisture content and freezing process for Areca catechu seeds. Seeds with 17-21 % moisture content and treated with PVS2 vitrification solution showed higher survival rates after cryopreservation. The oxidative markers of A. catechu seed embryos exhibited alterations due to vitrification freezing. However, there was no substantial association between seed viability and oxidative markers, such as reactive oxygen species (ROS), suggesting that oxidative damage mediated by ROS is not the primary factor contributing to the diminished viability of A. catechu seeds following cryopreservation. The inclusion of vitamin E, reduced glutathione (GSH), and 2-nitrobenzoic acid in the vitrification cryoprotectant achieved some improvement in seed viability of approximately 10 times, with GSH exhibiting the most pronounced optimizing effect.