As for many other taxa, hummingbird diversity declines away from the equator, but the causes for this decline are still disputed and might involve, among others, climatic factors or the availability of food resources. Because hummingbirds are one of the classical examples for plant–animal coevolution, it has been proposed that the diversity of hummingbird assemblages might depend on the diversity of food plants available. We tested this hypothesis by studying the hummingbird assemblages and their food plants for 1 year at six sites along a 660-km-long transect in Bolivian lowland forests extending from the southernmost Amazonian rain forests to dry Chaco forests. Hummingbird diversity was higher in the northern three sites as compared to the southern ones, with an abrupt decline in species numbers and a corresponding change in taxonomic composition at the boundary from evergreen to drought deciduous forests. Hummingbird diversity and abundance were only weakly correlated to climatic factors or to the diversity of humming-visited flowers, but strongly to the seasonal abundance of flowers. The overlap in nectar diet between hummingbird species depended on the number of plant species: when numerous species were available, the hummingbirds segregated by feeding preferences, but when few flowers were available, all hummingbirds fed on the same plants. We conclude that the local diversity of hummingbird species is not primarily determined by the diversity of food plants, but rather by the abundance of flowers available at any given point in time.