ABSTRACT Alstroemeria is a genus native to South America and commercially has commonly been propagated vegetatively by rhizome division, with low efficiency, high time consumption and a high risk of virus dissemination. In vitro propagation has several advantages, particularly in terms of efficiency and has been applied to the micropropagation of alstroemeria. This study aims to describe an efficient method for the in vitro propagation of Alstroemeria pallida Graham, a Chilean native species of high ornamental value. Concentrations of agar (0.0, 3.5 and 7.0 g L-1) and 6-benzylaminopurine (BAP) (0.0, 0.5, 1.0 and 2.0 mg L-1) were supplemented with MS culture medium to evaluate explant weight (g), rhizome length (cm), shoot length (cm) and proliferation rate. The highest explant weight was observed in rhizomes grown in culture medium supplemented with 3.5 g L-1 agar (3.79 g), and treatments using 2.0 g L-1 BAP showed the highest weight increase (3.33 g) after 8 wk. The proliferation rate rose with increasing concentrations of BAP, whereas low concentrations of BAP promoted longer shoots. An efficient method for in vitro propagation of A. pallida rhizomes was described, which could be useful for its conservation, domestication and further breeding.