This study presents a standardized protocol for the cryopreservation of Rauvolfia serpentina pollen, ensuring the long-term conservation of its genetic material for breeding and conservation initiatives. Rauvolfia serpentina, commonly known as Indian snakeroot, is a valuable medicinal plant renowned for its alkaloid content, particularly reserpine, which is widely used in the treatment of hypertension and mental disorders. The protocol includes systematic steps for pollen collection, desiccation, ultra-low temperature storage in liquid nitrogen (−196 °C), and post-thaw viability assessment through in vitro germination and pollination tests. This approach provides a reliable method for maintaining pollen viability and fertility, supporting future research and sustainable cultivation efforts.