Leaf anatomical and micromorphological traits are key taxonomic markers in the Bambusoideae (Poaceae). Otatea, the second most diverse genus in the subtribe Guaduinae, comprises 13 species distributed from Mexico to Colombia, exhibiting remarkable ecological adaptability from tropical dry to moist cloud forests. We hypothesized that species adapted to xeric versus mesic environments would show distinct morphological signatures, and tested this by analyzing the leaf anatomy and micromorphology of 11 species using tissue clearing and embedding techniques (Paraplast (R) and methyl acrylate) on samples collected in Mexico (2020, 2022). While the Stomatal Index (SI) showed only non-significant correlations with bioclimatic variables, we identified several key diagnostic traits for species differentiation. These included variations in papillae abundance, prickle morphology, the presence of micro- and macrohairs, as well as cross-sectional differences in rib shape, vascular bundle number, and mesophyll organization-against a background of consistent features like stomata, bulliform cells, and prickles. This comprehensive characterization provides valuable taxonomic tools and highlights the structural basis of the genus's ecological breadth.