Orchids are one of the most commercially valuable plants produced and sold worldwide. Many of them are with ornamental qualities such as those of the Vanilla genus, which contains over 100 species found worldwide, but many of them occurring in Mexico are endangered and need measures for their conservation. Minimal growth in vitro has proven to be very effective in this regard. However, its use necessitates the optimization of the most appropriate conservation protocol. As a result, it was suggested that this study be conducted with the goal of evaluating the effect of various LED light illumination spectra (blue light: 400-500 nm, red light: 700-800 nm, white light: 400-450 nm, and a combination of red and blue light: 660 and 460 nm, respectively, as well as a fluorescent light control: 400-450 nm) in the in vitro preservation process of four Vanilla genotypes. The results showed that the treatment with blue LED light was the most effective (V. planifolia wild morphotype (1.9 ± 0.0), interspecific hybrid of V. planifolia × V. pompona (1.4 ± 0.1), V. planifolia RH₃₅₀ (1.6 ± 0.1), and for the V. planifolia ʽMansaʼ (1.8 ± 0.1)). The results obtained showed that there was a reduction in the growth of vanilla shoots after six months of cultivation under the blue LED light spectrum. Consequently, the blue LED spectrum is appropriate for the slow-growth in vitro conservation of Vanilla spp. germplasm.