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Content of bioactive saponins of <i>Medicago</i> species suggests that they may also exert, as previously demonstrated on <i>M. sativa</i>, nematicidal properties exploitable for the formulation of new products for sustainable phytoparasitic nematode management. This study was addressed to highlight the bioactivity of saponins from five different <i>Medicago</i> species still poorly known for their biological efficacy, i.e., <i>M. heyniana, M. hybrida, M. lupulina</i>, <i>M. murex</i> and <i>M. truncatula</i>, against the plant parasitic nematodes <i>Meloidogyne incognita</i>, <i>Xiphinema index</i> and <i>Globodera rostochiensis</i>. The bioactivity of the extracts from the five <i>Medicago</i> species was assessed by in vitro assays on the juveniles (<i>J2</i>) and eggs of <i>M. incognita</i> and <i>G. rostochiensis</i> and the adult females of <i>X. index</i>. The suppressiveness to <i>M. incognita</i> of soil treatments with the <i>Medicago</i> plant biomasses was also investigated in a tomato experiment. The nematicidal activity of the five <i>Medicago</i> species was reported and discussed in relation to their phytochemical profile.