An efficient whole plant regeneration method from callus cultures of Piper auritum was achieved through organogenesis derived from leaf tissue. Proliferating callus and shoot cultures derived from leaf tissue explants placed on Murashige and Skoog (MS) medium supplemented with 2.0 mg·L–1 2, 4-dichlorophenoxyacetic acid (2,4-D) plus 1.5 mg·L–1 kinetin. Optimum combination of hormones (mg·L–1) for shoot induction was 0.5 2,4-D: 1.5 mg·L–1 kinetin (by volume), that resulted in a high rooting rate (49.6 shoots per explant). All of the plants elongated when using a medium consisting of 0.1 mg·L–1 2,4-D plus 1 mg·L–1 kinetin. Elongated shoots were successfully rooted (100%) on half-strength MS medium supplemented with 2.0 mg·L–1 indole-3-acetic acid. All plantlets survived to the growing conditions of a greenhouse. This study demonstrates that leaf tissue of P. auritum is competent for adventitious shoot regeneration and establishes an efficient and useful protocol for the multiplication and conservation of P. autirum for further investigation of its medicinally active constituents.