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Tracer‐aided studies to understand plant water uptake sources and dynamics in tropical ecosystems are limited. Here, we report the analysis of dry season source water uptake patterns of five unique ecosystems of Costa Rica across altitudinal (<150–3,400 m asl) and latitudinal (Caribbean and Pacific slopes) gradients: evergreen and seasonal rainforests, cloud forest, Páramo and dry forest. Soil and plant samples were collected during the dry season in 2021. Plant and soil water extractions were conducted using centrifugation. Stem water extracted volume and stem total water content were calculated via gravimetric analysis. Water source contributions were estimated using a Bayesian mixing model. Isotope ratios in soil and stems exhibited a strong meteoric origin. Enrichment trends were only detected in stems and cactus samples within the dry forest ecosystem. Soil profiles revealed nearly uniform isotopic profiles; however, a depletion trend was observed in the Páramo ecosystem below 25 cm. More enriched compositions were reported in cactus samples for extracted water volumes above ~20% (adj. r2 = 0.34, p < 0.01). The most prominent dry season water source in the evergreen rainforest (74.0%), seasonal rainforest (86.4%) and cloud forest (66.0%) corresponded to well‐mixed soil water. In the Páramo ecosystem, recent rainfall produced by trade wind incursions resulted in the most significant water source (61.9%), whereas in the dry forest, mean annual precipitation (38.6%) and baseflow (33.1%) were the dominant sources. The latter highlights the prevalence of distinct water uptake sources between recent cold front rainfall (near‐surface soil storage) to more well‐mixed soil moisture during the dry season, revealing ecohydrological processing previously unknown in this tropical region. [ABSTRACT FROM AUTHOR]