Developing Cryopreservation Methods of Wheat Roots

In the midst of record breaking rates of plant species extinction due to climate change and fungal diseases, a universal cryopreservation method would provide a means for preservation of these many different species. The concept of plant root cryopreservation first emerged in the late 1960’s, and with it came new avenues of preserving tissue for the purposes of agriculture and research. Frozen tissues can be transported and stored more reliably than other more conventional means. When thawed, they have the potential to be cultured and grown. Several different methods of cryopreservation exist. This experiment used the Fast (3°C/minute) and Slow freeze (0.3°C/minute) method with a controlled freezing unit on a wheat plant species. Cryoprotectant solutions containing 10% DMSO or 10% glycerol were used in conjunction with high (3.11 M) and low (1.5 M) sorbitol concentrations. A no sorbitol group was also tested. After being frozen, samples were transferred to a liquid nitrogen tank for storage, and later thawed. Thawed samples were stained with fluorescent dyes to observe live and dead cells under fluorescent microscopy. Root tips were deemed “surviving” if multiple live nuclei were present. In the Glycerol group, Slow freezing showed a higher survival rate, while the DMSO group had good survival rates for both freezing rates. The Slow freeze rate showed greater survival rates overall compared to the Fast rate. Future experiments to develop a universal method will include testing on various plant species, such as potato plants, and succulents