Background: Palikur Amerindians live in the eastern part of French Guiana which is undergoing deep-seated changes due to the geographical and economic opening of the region. So far, Palikur's traditional ecological knowledge is poorly documented, apart from medicinal plants. The aim of this study was to document ethnobotanical practices related to traditional construction in the region. Methods: A combination of qualitative and quantitative methods was used. Thirty-nine Palikur men were interviewed in three localities (Saint-Georges de l'Oyapock, Regina and Trois-Paletuviers) between December 2013 and July 2014. Twenty-four inventories of wood species used in traditional buildings were conducted in the villages, as well as ethnobotanical walks in the neighboring forests, to complete data about usable species and to determine Linnaean names. Results: After an ethnographic description of roundwood Palikur habitat, the in situ wood selection process of Palikur is precisely described. A total of 960 roundwood pieces were inventoried in situ according to Palikur taxonomy, of which 860 were beams and rafters, and 100 posts in 20 permanent and 4 temporary buildings. Twenty-seven folk species were identified. Sixty-three folk species used in construction were recorded during ethnobotanical walks. They correspond to 263 botanical species belonging to 25 families. Posts in permanent buildings were made of yawu (Minquartia guianensis) (51%) and wakap (Vouacapoua americana) (14%). Beams and rafters were made of wood from Annonaceae (79%) and Lecythidaceae (13%) families. The most frequently used species were kuukumwi priye (Oxandra asbeckii), kuukumwi seyne (Pseudoxandra cuspidata), and pukuu (Xylopia nitida and X. cayennensis). Conclusions: Although the Palikur's relationship with their habitat is undergoing significant changes, knowledge about construction wood is still very much alive in the Oyapock basin. Many people continue to construct traditional buildings alongside modern houses, using a wide array of species described here for the first time, along with the techniques used.