This chapter explores the activities and strategies for preserving plant biodiversity at the National Gene Bank of Tunisia (NGBT). As the guardian of the national germplasm, the NGBT implements advanced conservation practices, including acquiring accessions, multiplying, regenerating, evaluating, and valorizing them. Special emphasis is placed on valorizing indigenous resources, to support the local economy and engage farmers in protecting genetic heritage. The chapter highlights the NGBT’s crucial role in preserving genetic diversity through protocols that adhere to international standards. Specific strategies are employed for allogamous plants, and demonstrating the proactive approach of the NGBT in maintaining genetic integrity. The in-depth evaluation of accessions, encompassing genetic, phenotypic, and agronomic aspects, underscores the NGBT’s dedication to efficient collection management. In terms of resource valorization, strategic partnerships have been established with various stakeholders, to stimulate the local economy. The NGBT positions itself as a cornerstone for preserving the national genetic heritage, benefiting biodiversity and the local economy. Additionally, operational aspects and documentation are central, with the utilization of the Grin-Global platform for collection management and the complete digitization of genetic resources. The NGBT actively participates in digitization, assigning Digital Object Identifiers (DOIs) to enhance traceability and integrating barcodes for efficient electronic resource management. This approach enhances operational efficiency while significantly contributing to the preservation of the national genetic heritage. Committed to addressing global challenges, the NGBT exemplifies its dedication to the richness of national plant biodiversity. This chapter provides an overview of the activities and strategies employed by the NGBT to preserve plant biodiversity, emphasizing its role in safeguarding the national genetic heritage and proactive measures to maintain genetic integrity. It covers various aspects such as acquisition, multiplication, regeneration, evaluation, and valorization of plant germplasm, highlighting the NGBT’s commitment to effective collection management and resource valorization. Additionally, it discusses the importance of strategic partnerships and digital documentation in enhancing operational efficiency and traceability.