Sour orange (Citrus aurantium L.) has a high nutritional and gastronomic value in Huacho, Lima, Peru, but is currently considered a species in danger of disappearing from the district and surrounding areas due to phytosanitary problems. For the recovery and repopulation of this species, biotechnological techniques were proposed to obtain pathogen-free plants. Therefore, the objective of the present research was to micro propagate sour orange in vitro from nodal segments. Sour orange nodal segments were disinfected in different concentrations of NaClO, then placed in MS culture medium (Murashige and Skoog) supplemented with BAP, KIN, and AG3 for the multiplication phase, and then transferred to MS medium supplemented with IBA and ANA for the rooting phase. The evaluation of the percentage of contamination was carried out after 10 days, the evaluation of shoot formation in the multiplication phase was carried out after 30 days, and the evaluation of rooting after 30 days. In the disinfection and in vitro establishment phase, 0% contamination and 0% oxidation of the explants were obtained. In the in vitro multiplication phase, the best results were obtained in the M8 culture medium, generating 4.7 shoots per explant. Finally, the E4 medium yielded 94.7% of rooted explants, 23.4 mm root length, and 2.2 roots per explant.