A cladistic phylogenetic analysis at generic level of the subfamily Amaurobioidinae is presented. The analysis is based on a dataset of 93 representative species scored for one behavioral and 199 morphological characters. Tree searches were made under equal and implied weights according to homoplasy, and the results were compared in terms of sensitivity to jackknife resampling. Mildest weighting functions produced trees more robust to resampling, and those results were selected as the working phylogenetic hypotheses. Groups of weak support as identified by jackknifing and Bremer indices are in general those that vary in resolution with different character-weighting schemes.Seven outgroup representatives were included (Malenella nana Ramírez, from Malenellinae, and six Anyphaeninae species). In this analysis Anyphaeninae, previously identified as sister group of Amaurobioidinae, is paraphyletic, but forcing its monophyly does not alter the groupings within Amaurobioidinae. The monophyly of the genera is in general well supported, but some particularly conflicting groups are discussed. In contrast, the relationships among genera are in general problematic.Amaurobioidinae is diagnosed by a pronounced indentation at the base of male palpal tegulum, and by a particular male copulatory bulb conformation, with a paramedian apophysis. The subfamily is classified in two tribes (Gayennini and Amaurobioidini); the genus Josa Keyserling, probably sister group to Gayennini, is not assigned to either tribe.The tribe Amaurobioidini is mainly diagnosed by an apical loop of the sperm duct in the male copulatory bulb. It includes 10 genera: Amaurobioides O.P.-Cambridge is restricted to seashores of southern continents. Clubiona chilensis Nicolet, transferred to Amaurobioides, is the first true record of the genus for South America. The male of Axyracrus elegans Simon, three species of Aysenia Tullgren, and three of Coptoprepes Simon are newly described. Four new genera are proposed in Amaurobioidini: Gamakia, Selknamia (described for one new species each), Aysenoides (for three new species), and Negayan (type species Gayenna tridentata Simon, including also Axyracrus coccineus Mello-Leitão, Clubiona paduana Karsch, Gayenna excepta Tullgren, Gayenna exigua Mello-Leitão, and Tomopisthes lebruni Simon). The previously revised genera Acanthoceto Mello-Leitão and Ferrieria Tullgren are also included in the tribe. The basal branch and most intergeneric branches of the tribe have low support values. Amaurobioides and Negayan, however, are relatively well supported.The tribe Gayennini is well defined by a homogeneous conformation of male and female genitalia, with a distinctive secondary conductor and spherical spermathecae. It includes 11 genera: Gayenna Nicolet includes only G. americana Nicolet from Chile and adjacent Argentina. Arachosia O.P.-Cambridge comprises many species previously assigned to Oxysoma. Abuzaida striata Keyserling, Anyphaena oblonga Keyserling, Gayenna proseni Mello-Leitão, Gayenna duplovittata Mello-Leitão, Gayenna bonneti Mello-Leitão, Oxysoma dubium Berland, Oxysoma bifasciatum Mello-Leitão, Oxysoma cubana Banks, Oxysoma polytrichium Mello-Leitão, Phidyle bergi Simon, and Samuza praesignis Keyserling are transferred to Arachosia. The males of Arachosia bergi (Simon), A. honesta Keyserling, and Arachosia praesignis (Keyserling) are newly described. Arachosia is easily recognized by the thick setae on the anterior lateral spinnerets, and it has good support values. A very diverse group of species here assigned to the genus Sanogasta Mello-Leitão is paraphyletic in terms of Arachosia. It includes many of the species formerly placed in Gayenna Nicolet. Anyphaena maculatipes Keyserling, Clubiona maculosa Nicolet, Gayenna paucilineata Mello-Leitão, Gayenna alticola Simon, Gayenna bonariensis Mello-Leitão, Gayenna rufithorax Tullgren, Gayenna x-signata Keyserling, Gayenna approximata Tullgren, Samuza minuta Keyserling, and Tomopisth