Investigation on karyotype, 4C nuclear DNA amount and interphase nuclear volume (INV) of different HimalayanBupleurum species belonging toUmbelliferae revealed genetic differentiation. Numerical and structural alternation of chromosomes in interspecific level were manifested in their statistically significant altered species specific 4C nuclear DNA content. Somatic chromosome number ranged between 2n = 14 and 2n = 16.B. himalayense was reported for the first time having 2n = 16 chromosomes. Correlation coefficient among the various chromosomal and nuclear parameters showed no significant progressive or regressive interdependence except in between INV and nuclear DNA amount. Critical differences between 4C DNA content showed interspecific variation.