A phytochemical investigation of Bidens procera L.C.Xu ex X.W.Zheng afforded two novel polyacetylenes, tridecane-2E-monoene-4,6,8-triyntylen-1,13-diol-12-O-β-glucoside (1) and tetradecane-2E,8E-diene-4,6-diyne-1,14-diol-13-O-β-glucoside (2), together with ten known compounds (3−12). Their chemical structures were elucidated by NMR and MS spectrums as well as the comparison of the published data. Furthermore, the chemotaxonomy of the yielded compounds was also discussed.