Genetic diversity allows plants to adapt to changing environmental conditions to survive and increases their ability to respond to yield, production, pests and diseases. The application of molecular markers developed due to developments in biochemistry, molecular biology, and plant technology has shed light on plant genetics and breeding studies and produced an enormous amount of knowledge. The theoretical knowledge will guide in determining the scope, amount, and distribution of different aspects of genetic diversity harbored in plants and how it is structured, determining what, where, and how to protect and management of the studies in practice. In plants, molecular markers have been used in the assessment of genetic diversity and population genetics, characterization of germplasm, investigation of phylogenetic relationships, identification of species, hybrids and varieties, ecology, evolutionary biology, taxonomy, selection and breeding studies based on molecular markers in the construction of gene maps and QTL maps in the last four decades. Each of the known molecular markers or their derivatives has different methodologies, advantages, or disadvantages. Comparative studies of different molecular markers performed in different plant species along with their wild and primitive relatives offer researchers the opportunity to determine and apply the most appropriate methodologies for future detailed studies. The sustainability of life on earth depends first on the genetic diversity in individuals, second on the species diversity in the ecosystems, and finally on the ecosystem diversity. Deterioration or loss in any of these will disrupt the balance between living things.