The present study was intended to produce a protocol for large scale production of callus and compare the phytochemical constituents and isoperoxidase profile of leaves and calli of Solanum anguivi Lam. Maximum percentage of friable callus (86.3 ± 0.94) proliferation was obtained on MS medium supplemented with 1.0 mg/l of 2, 4-D. The phytochemical study revealed that the high degree of steroids, alkaloids, phenolic compounds, flavanoids, tannins present in leaves derived calli and calli induced secondary calli. A total of 23 bands in seventeen different positions with seven activity regions (PRX1-7) were observed in the isoperoxidase enzyme system of Solanum anguivi. The leaves showed their uniqueness by the presence of following bands MW-Rf – 0.1250, 0.2125, 0.3250, 0.5000, 0.5750 and 0.6500. The primary calli expressed their individuality by the occurrence of MW-Rf 0.0875, 0.2875, 0.3625 and 0.5250. The MW-Rf 0.0750, 0.4500, 0.4875 and 0.5875 are present only in secondary calli (Calli mediated calli). We observed the genetical difference indicates the possibility for the differences due to the morphogenesis activity of the plant not because of chemical constituents.