The Andean mortiño (Vaccinium floribundum Kunth) grows wild in the northern paramos of South América. The berries present potential opportunities for agribusiness, but an efficient technology for the production of plants and berries is required. On the other hand, the development of plant production methods from mortiño seeds will allow the plants’ accessibility and its potential use in breeding programs. The objective of this study was to develop an efficient in vitro protocol for accelerated production using seeds. We carried out the research in four phases: seed germination, plant multiplication, rooting, and acclimatization in the greenhouse. For in vitro seed germination, we studied the effect of two culture media [woody plant medium (WPM) and Murashige and Skoog medium (MS], two photoperiods (16 and 24 hours of light), and two temperatures (18 and 28 °C). The best treatment was found to be WPM + 24 h light + 18 °C. In the micropropagation of seedlings, the effect of two concentrations of three cytokinines [(trans-zeatin riboside (TZR), zeatin (ZEA), and 2-isopentenyl adenine (2iP)], and two photoperiods (16 and 0 h light) was evaluated, whereby the concentration of 0.5 mg L−1 of TZR was the best treatment. For rooting, two doses of three auxins [indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), and indole-3-acetic acid (IAA)] were evaluated, resulting in the 2 mg·L−1 concentration of IBA giving the best root induction. Finally, the in vitro rooted plants were acclimatized in a greenhouse. We found that peat was the best substrate. These results show that the technology developed here is useful for in vitro production of V. floribundum using seeds.

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