A study concerning the bee fauna of the Serra do Itajaí National Park, state of Santa Catarina, was carried out during ten months in the domain of the Atlantic Rainforest. Bees were collected monthly between October 2006 and December 2007, in two sampling localities, from 8:00 h to 16:00 h, totaling 151 h of sampling. Bees were captured with entomological nets in flowers, during flight, on the ground or while foraging for sweat on collectors. A total of 1,616 individuals were collected (1,240 females and 376 males), distributed in 89 species, 46 genera, 16 tribes and five subfamilies. Apinae was the richest subfamily with 47 species. The genera with the greatest number of species were Augochloropsis Cockerell, 1897 (nine), Ceratina Latreille, 1802 (nine) and Dialictus Robertson, 1902 (seven). Paratetrapedia fervida (Smith, 1879) was the most abundant species, comprising 11.57 % of the sampled individuals