The present study was aimed to regenerate plants from internode explants and analyze the origin and developmental pattern of cells during different developmental stages of Solanum trilobatum L. To analyze and compare the developmental stages were assessed from callus, organogenic callus, multiple shoots, and root. The best response of callus and organogenic callus induction (78.4% and 65.2%) was obtained on 3.0 mg/l of 2,4-D (2,4-dichlorophenoxyacetic acid) supplemented in Murashige and Skoog’s (MS) medium. Organaogenic callus induction was maximum (81.2%) in medium supplemented with 3.0 mg/l of BAP (6-benzylaminopurine) + 1.0 mg/l of NAA (α-naphthaleneacetic acid) in different combinations. A higher rate of multiple shoot induction (80.1%) was observed on medium supplemented with 3.0 mg/l BAP with 1.0 mg /l of NAA. For best root initiation (63.5%) was induced on medium containing 1.0 mg/l of IBA (indolebutyric acid). In vitro plantlets regeneration was successfully achieved in maximum (80.8%) medium supplemented with BAP 2.0 mg/l with NAA 1.0 mg/l and acclimatized in vermisulite:sand:soil (1:1:1) mixture and carefully transferred to field condition they showed 90% of survival. Histological analysis revealed that the shoots were originated directly on the proximal cut surface and root induction was distal cut end of internode.