This study tested three hypotheses regarding how plants respond to the spatial heterogeneity in light availability in the rain forest understory: (1) understory plants occur preferentially in the lighter parts of the understory; (2) understory palms are more shade tolerant than other understory plants; (3) rain forest plants differ in their ontogenetic response to understory light conditions. The study was carried out in old-growth rain forest in the Yasuní National Park, Amazonian Ecuador. The hypotheses were tested by comparing the distributions of 20 plant species (1454 individuals) over microsites with differing degrees of exposure to canopy gaps to the background distribution of these microsites in the forest. The gap exposure of a given microsite was described by an index based on the number and size of gaps in the canopy to which the site was exposed. Two plant height classes were studied: 0.80–2.49 and 2.50–5 m. The first and third hypotheses were accepted, while the second hypothesis was rejected. The results for the individual species corresponded well with what is known from earlier studies about the ecology of these species or close relatives, suggesting that the patterns observed can be generalized for Neotropical rain forests. Notably, the most abundant species in the study represent several different life history strategies. Thus, abundance in the rain forest understory can be achieved by several different strategies. This suggests that niche differentiation in terms of response to small changes in understory light conditions may be an important factor in the maintenance of the high local plant species richness of tropical rain forests.