The International Union for Conservation of Nature (IUCN) estimates that approximately 10% of the world’s vascular plant species, or 20,000–25,000 species, are threatened. There is a global appeal for emergency conservation efforts to safeguard plant diversity and ensure a sustainable future. To address this challenge, both natural and biotechnological solutions are being explored to conserve plant genetic resources, with germplasm playing a critical role. Plant genetic resources encompass all hereditary material found in crops and their wild relatives, critical for farming resilience and profitability. Conserving genetic diversity is critical for long-term food security, as environmental problems and genetic erosion endanger these precious resources. The goal is to preserve a broad intraspecific genetic variety for present and future usage. This chapter will emphasize on the expanding significance of tissue culture and biotechnology in conserving germplasm and assisting breeders, which is critical for global agricultural development and adjusting to changing problems endangering food security. A key aspect of tissue culture is the in vitro growth of plant cells, tissues, or organs under sterile conditions. In tissue culture, the use of plant growth regulators facilitates the micropropagation of rare or endangered species and improves genetic modification processes. This process helps with the propagation of rare or endangered species, disease-free planting material, and fast plant multiplication. Genetic material is stored at −196 °C during cryopreservation, preserving tissues, cells, and seeds. Cryopreservation maintains the genetic integrity of seeds, tissues, and cells, guaranteeing long-term conservation and guarding against genetic loss. Genetic variety is preserved for breeding, study, and restoration through gene and seed banks, offering an economical way to preserve genetic resources. To maintain plant genetic resources and seed germplasm, an integrated method utilizing tissue culture, cryopreservation, gene banks, and seed banks is used. This technique supports food security, resilient ecosystems, and sustainable agriculture by safeguarding genetic diversity against habitat loss and environmental change. As the world’s population and environmental concerns expand, advancements in tissue culture, cryopreservation, gene banks, and seed banks are critical to maintaining plant genetic resources and food security.