The Neotropical “club-tailed” scorpions of the genus Rhopalurus Thorell, 1876, and two related genera in family Buthidae C.L. Koch, 1837, i.e., Physoctonus Mello-Leitão, 1934, and Troglorhopalurus Lourenço et al., 2004, are revised, based on a simultaneous phylogenetic analysis of 90 morphological characters and 4260 aligned DNA nucleotides from three mitochondrial and two nuclear gene loci. The monophyletic New World buthid subfamily Rhopalurusinae Bücherl, 1971, to which these scorpions were originally assigned, is redefined, revised diagnoses and a key to identification of its genera and species (except for Centruroides Marx, 1890) provided, and their distributions mapped. The paraphyly of Rhopalurus Thorell, 1876, which comprises several monophyletic groups congruent with its disjunct distribution, justifies its relimitation and that of Troglorhopalurus Lourenço et al., 2004, the revalidation of Heteroctenus Pocock, 1893, and creation of Ischnotelson, gen. nov. (type species: Rhopalurus guanambiensis Lenarducci, Pinto-da-Rocha and Lucas, 2005) and Jaguajir, gen. nov. (type species: Rhopalurus agamemnon C.L. Koch, 1839). Ten new combinations are proposed: Heteroctenus abudi (Armas and Marcano Fondeur, 1987), comb. nov.; Heteroctenus bonettii (Armas, 1999), comb. nov.; Heteroctenus garridoi (Armas, 1974), comb. nov.; Heteroctenus gibarae (Teruel, 2006), comb. nov.; Heteroctenus princeps (Karsch, 1879), comb. nov.; Ischnotelson guanambiensis (Lenarducci, Pinto-da-Rocha and Lucas, 2005), comb. nov.; Jaguajir agamemnon (C.L. Koch, 1839), comb. nov.; Jaguajir pintoi (Mello-Leitão, 1932), comb. nov.; Jaguajir rochae (Borelli, 1910), comb. nov.; Troglorhopalurus lacrau (Lourenço and Pinto-da-Rocha, 1997), comb. nov. Three new species are described: Ischnotelson peruassu, sp. nov. (type locality: Parque Estadual do Peruassu, Minas Gerias, Brazil); Physoctonus striatus, sp. nov. (type locality: Castelo do Piauí, Piauí, Brazil); Rhopalurus ochoai, sp. nov. (type locality: San Agustín, Edo. Zulia, Venezuela). Fifteen new junior subjective synonyms are proposed: Rhopalurus acromelas Lutz and Mello, 1922, Rhopalurus melleipalpus Lutz and Mello, 1922, Rhopalurus iglesiasi Werner, 1927, Rhopalurus lambdophorus Mello-Leitão, 1932, Rhopalurus dorsomaculatus Prado, 1938, and Rhopalurus goiasensis Prado, 1940 = Jaguajir agamemnon (C.L. Koch, 1839); Rhopalurus pintoi kourouensis Lourenço, 2008 = Jaguajir pintoi (Mello-Leitão, 1932); Rhopalurus crassicauda Caporiacco, 1947, Rhopalurus amazonicus Lourenço, 1986, and Rhopalurus crassicauda paruensis Lourenço, 2008 = Rhopalurus laticauda Th