The present article has reported a detailed karyotype analysis and 4C nuclear DNA estimation in different cultivars and populations of Clitoria ternatea L. of Fabaceae. This plant, widely distributed in tropical regions in India, is a climber and is an economically important medicinal plant. The karyotype study has revealed a constancy in the diploid somatic chromosome number (2n=16) in all these cultivars and populations. However, the nature and number of nucleolar chromosomes varied. The centromeric chromosomes were either metacentric or sub-metacentric in all these cultivars. The total chromosome length and volume differed among these cultivars and populations having the same somatic chromosome number. The differential condensation of chromosomes and the association of variable amounts of proteins in chromosome composition might have been responsible for such differences. In situ cytophotometric studies revealed an intermediate genome size in this species. This is the first report on the detailed karyotype and genome size of cultivars of Clitoria ternatea L. © 2016 The Japan Mendel Society.