The past two decades have brought with them remarkable progress in plant chromosomal research. The chromosome structure has been clarified in great detail, enabling identification of gene sequences at the microscopic level, which has aided the analysis of biodiversity. Knowledge of chromosome structure has played a crucial role in the improvement of crop species and has far-reaching implications. The manipulation and engineering of chromosomes involves a panoply of novel methods, combining conventional and modern techniques of biotechnology. A working knowledge of such techniques is essential for today’s students and researchers, and the plant system, because of totipotency, requires special treatment. This treatise covers all the latest methods involved in the study of evolution, biodiversity, chromosome manipulation and engineering.