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Background. Tuberose (Polianthes tuberosa L.) is a native plant of central and southern Mexico, cultivated for cut flower and as for the perfume industry. Farmers propagate it vegetatively, so it is important to generate new varieties by implementing efficient techniques, both to induce mutations and micro-propagation. Objective. Establish the protocol for the in vitro propagation of P. tuberosa L. from foliar and flower tissue. Methodology. Leaf and button explants were placed on the surface of the culture medium base GC, containing inorganic salts Murashige and Skoog (1962) per liter, coconut water 50 mL L-1, 20 g L-1 of sucrose, 6.4 g L-1 of agar, being the pH adjusted to 5.7. With this basic medium were prepared a series of culture media containing benzylaminopurine (BAP) , naphthaleneacetic acid (ANA), 2,4-dichlorophenoxyacetic acid (2,4-D), indole acetic acid (IAA) and kinetin. Results. Shoot regeneration (3.5 %) was solely from button tissue explants on solid media containing BAP and ANA; media T1 (1.1 %), T3 (3.5 %) and T11 (2.5 %). The shoots developed roots in the medium with indolbutiric acid (1 mg L-1) and BAP (0.1 mg L-1). In foliar tissue cultures, the regeneration of green callus was achieved in a higher percentage (50.9 %) in the solid media with BAP (4.5 mg L-1) and ANA (0.05 mg L-1) and sucrose 20 g L-1. Implications. The protocol was developed that allowed obtaining tuberose seedlings from flower buds. This attributes to the in vitro conservation of germplasm, and initiating genetic improvement programs for P. tuberosa with new and desirable characteristics, to increase biodiversity in the crop. Conclusions. Both shoot induction and development were observed in media containing BAP (4.5 mg L-1) and ANA (0.1 mg L-1).               .