Medicinal plants have increasing worldwide potential in pharmacological therapies, and their secondary metabolites are effective, with no adverse effects on the body. Technologies for plant cell culture have proven to be useful for both researching and generating plant secondary metabolites in vitro, and phytochemicals are reported to defend against abiotic stresses. This study employed four different types of abiotic elicitors (UV, NaCl, GA3, and Pt) to examine the relationship between abiotic stresses and the generation of secondary metabolites from Datura stramonium, D. innoxia, Solanum nigrum, and S.dubium. After breaking seed dormancy, Callus tissues were derived from two organs: the seeds and leaves of all the four species. In callus tissue, the high mean total alkaloid recorded for leaves was observed with values in S.dubium˂D.stramonium˂ D.innoxia˂ S.nigrum, while the seed values were scored as S.dubium˂ D.innoxia ˂ S.nigrum ˂D.stramonium. The resulting alkaloids from the callus treated with different elicitors showed that high alkaloid levels obtained from S.nigrum seeds treated with GA3 (1082.02 μg/ml) and the lowest alkaloid levels (58.17 μg/ml) were observed in leaves treated with platinum and in controlled D.stramonium seeds. Seed explants had more alkaloid production than leaves in all species. Atropine, a tropane alkaloid, was detected in 13 samples using GC/MS. D.innoxia seeds treated with NaCl, and had the highest atropine concentration (18.82ppm).