In the field of agrobiotechnology, we analyze and characterize plant biodiversity through cytological techniques such as flow cytometry and molecular cytogenetics, as well as by nucleic acid sequencing and genotyping of plant species and populations. Cytological approaches allow the determination of DNA content and ploidy levels. Sequencing of plant genomes and their genotyping reveal the genetic variability of a species and enable genotype–phenotype associations through genome-wide association studies (GWAS). This facilitates the identification of genes of interest to be introduced into elite agronomic lines via conventional or innovative breeding.