This study reported the effect of interaction between cytokinins and auxins to enhance accumulation of alkaloids in White henbane (Hyoscyamus albus L.). Plants of this specie were grown under controlled conditions and treated with plant-hormones: Auxins by: 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and 3-Indole Acetic Acid (IAA), Cytokinins by: Kinetin (K) and Benzyl Amino Purine (BAP), at 0-10 and 20 mg L(-1) rates isolated and interacted. The results showed that treatment of 2, 4-D and K at the highest applied rates 20 mg L(-1) increased the accumulation threefold rate estimated to 2.321% in the root plant part and 1.702% in the aerial plant part with the same plant-hormones but dosage of (20x10 mg L(-1)) in order. The TLC for alkaloid extracts shows that H. albus L. contains 6 alkaloids. In this study, it was concluded that the treatment with interaction of (Kx2,4-D) (20x20 mg L(-1)) gives the highest percent of alkaloids in the root and shoot parts compared to plant-hormones separated. The in vitro antibacterial activity was determined on microorganisms: Pseudomonas stutzeri, Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae. And performed by disc diffusion assay. Respectively, ethanol extracts showed no inhibitory effect on the microorganisms, however alkaloid extracts of H. albus L. of the same treatments with plant-hormones of shoot and root parts showed antibacterial activity against microorganisms that were tested. The results obtained in the present study suggest that alkaloid of H. albus L. can be used in treating diseases caused by the test organisms.