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The exploitation of biodiversity is essential to select resilient genotypes for sustainable cropping systems as one of the main challenges for plant breeding. Mapping traits of agronomic interest in specific genomic regions appears as another pivotal effort for the future development of novel cultivars. For this purpose, there is evidence that MAGIC and other exotic populations will play a major role in the coming years in allowing for impressive gains in plant breeding for developing new generations of improved cultivars. This Special Issue focused on the application of advanced technologies devoted to crop improvement and exploit the available biodiversity in crops. In detail, next-generation sequencing (NGS) technologies supported the development of high-density genotyping arrays for different plants included in this issue.