Accumulation of soluble sugars, heat-stable proteins and dehydrins in cryopreservation of protocorm-like bodies of Dendrobium candidum by the air-drying method

Protocorm-like bodies (PLBs) of Dendrobium candidum were successfully cryopreserved by the air-drying method. The optimal water content before freezing seemed to be at the range of 0.1 g H2O/g DW (11% on fresh weight basis) to 0.5 g H2O/g DW (33% on fresh weight basis). Changes in soluble sugars, heat-stable proteins and dehydrins during desiccation of PLBs were analyzed. Extensive accumulation of soluble sugars was observed at water content of about 7.2 g H2O/g DW (after 24 h desiccation), and the sugars content did not increase further during the following desiccation. The amount of heat-stable protein increased significantly when water content decreased to 1.0 g H2O/g DW (after approximately 66 h desiccation). Results from immunological detection showed that two bands of the heat-stable proteins with respective molecular masses of 28.7 and 34.3 kDa were dehydrins which appeared when water content dropped to 1.0 g H2O/g DW. Therefore, it seemed that accumulation of dehydrins happened later than that of soluble sugars. Interestingly, exogenous ABA treatment of PLBs before desiccation could also induce the accumulation of soluble sugars, heat-stable proteins and dehydrins. The possible roles of these substances in the acquisition of dehydration and freezing tolerance were discussed.