Polyphenols are a large diversity of chemical types and interactions that can be responsible for a multiplicity of protective functions ranging from toxicity and light/UV shielding to signal transduction. Bacharis antioquensis has been described as a potential source of new photoprotective compounds with antioxidant capacity associated to polyphenols compounds. The aim of the present work was to develop a micropropagation protocol of B. antioquensis and evaluate the production of polyphenols by in vitro plants exposure to UVB radiation. Branches in juvenile stage of B. antioquensis were collected, desinfected and cultured on half strength Murashige and Skoog medium, supplemented or not with growth regulators (TDZ, BA or GA3) on light/darkness conditions and liquid/solid media. After UV treatments, the absorption coefficient in the UVA-UVB range, the antioxidant capacity and the total phenol content (TPC) from all tissue cultures and the wild tissue were evaluated. Growth regulators, light conditions and type of culture medium (solid or liquid) had a favorable effect on the response of explants. Treatments containing BA + GA3 regulators (2 and 0.5 mg/L respectively) and TDZ (0.5 mg/L) showed positive results in bud growth in liquid medium and darkness. Results showed that UVR exhibited promoting effects on the accumulation of polyphenols, enhancing the absorption coefficient in the UVA-UVB range, the antiradical capacity and the TPC of B. antioquensis in vitro plants. Rev. Biol. Trop. 66(2): 754-764. Epub 2018 June 01.