Cryopreservation is one of the significant biotechnological approaches that has conserved plant genetic resources destroyed by habitat loss, climate change, and overexploitation. The method ensures long-term preservation of plant tissues, cells, and seeds at ultra-low temperatures that halt biological degradation and keep genetic materials safe for further use. Cryopreservation is a method that does not assimilate the characteristics of any other conservation methods, for its genetic materials are maintained in a stable form and hence apt to preserve rare and endangered species. It also assists with in situ conservation when it controls diseases and pests and acts as a gene bank, increasing diversity in reproductive techniques. Cryopreservation has recently been significant for sustainable food security by conserving the crop genetic diversity required to adapt to the agricultural challenges posed by climate change. Cryopreservation is one of the critical components for future directions, which will further support the preservation of genetic resources necessary for ecosystem restoration and innovative agriculture. This chapter highlights cryopreservation techniques, applications, and their vital role in plant genetic resource conservation, biodiversity, and sustainable resource management.