AbstractWe reviewed the status of orthodox seed collections of globally threatened plants conserved in − 20 °C long-term storage at the Millennium Seed Bank, Royal Botanic Gardens, Kew, UK in terms of their geographic and bioclimatic representativeness, taxonomic and genetic diversity, quality and physiological status. The comprehensive dataset used spans over 45 years of worldwide conservation effort across various organisations. The data provides evidence-based results and future directions for the represented globally threatened flora that are of relevance to all plant conservation and seed banking organisations across the globe. The reviewed sample includes 523 collections and represents a wide geographic range, originating from 67 countries, from all nine bio-geographic continents. The majority of collections originated from temperate climates and from habitats with no dry seasons but experiencing warm summer periods. The taxonomic composition of the collections highlighted a substantial diversity, with 303 taxa (four extinct in the wild; 56 critically endangered; 105 endangered; and 138 vulnerable) represented by 297 species, 199 genera and 74 families. Almost four fifths of the collections were harvested from wild habitats. Whilst wild-origin collections can harbour useful genes not available in the cultivated gene pool, for threatened plants both collections and taxa are likely to suffer from low genetic diversity as a low number of individual plants, populations and/or potentially viable or usable seeds were sampled at the original harvest. Large numbers of empty and infested seeds in the original harvest have significantly affected the quality of collections in terms of availability of potentially viable or usable seeds in collections. As a result, just over one third of taxa and one fifth of collections consisted of ≥ 5000 potentially viable or usable seeds. Viable seeds exhibited a sound physiological status in terms of germinability and viability at the initial round of germination tests after storage, but on average, relative germination and viability achieved were below 85%. A decline in germinability during their variable time of storage was evident for 16% of the 78 collections analysed for longevity. According to a set of criteria, suitable germination protocols for propagation of plants from seeds were identified for 165 taxa. Given the apparent differences between wild species, especially those that are rare and threatened, and domesticated crops, the quality and physiological status of reviewed collections are reasonably sound. The characteristics we observed for collections, the challenges we identified for conserving them and the germination protocols we suggested for propagation of plants from seeds have the scope to be noted, integrated and used globally across various conservation activities and policies.