Chile is part of one of the centers of crop origin identified by Vavilov, e.g., for strawberries and potatoes. It is also a center of diversification of other crop species such as maize, beans and quinoa. It is one of the biodiversity hotspots of the world and several native species have potential for domestication. All of these types of species are considered Plant Genetic Resources for Food and Agriculture (PGRFA). However, the rich plant genetic diversity present in Chile is being lost, mostly due to human activity. Therefore, ex situ and in situ conservation of this diversity are of critical importance. In this review we show the achievements in PGRFA conservation activities in the last 15 yr and in plant breeding for the last 60 yr in this country. Several gene banks exist, administrated by different institutions, with over 48 000 accessions preserved, mostly cereals (65%) and grain legumes (23%). Significant advances were achieved between 2006 and 2020 in the conservation, regeneration, characterization and documentation of PGRFA, but work is still needed to complete a fully operable data base for all collections. Over 16 000 accessions of Chilean origin are also kept in gene banks abroad. Plant breeding programs of several agriculturally important crops have made an outstanding contribution to Chilean agriculture and food security, with more than 375 commercial cultivars developed. More effort needs to be made to strengthen ex situ conservation and the sustainable use of PGRFA under coordinated actions, guided by a national strategy on genetic resources, if significant contributions are to be made in response to climate change.