Expression of Tenod40, a tobacco (Nicotiana tabacum cv. Xanthi nc) homologue of Msenod40, a gene expressed early during alfalfa (Medicago sativa) nodule development, was significantly and consistently increased (about three-fold) in tobacco roots of plants grown in sand and colonized by the arbuscular mycorrhizal (AM) fungus Glomus intraradices, as compared to non-mycorrhizal controls. In alfalfa (Medicago sativa cv. Gilboa), a similar induction of Msenod40 expression was also detected in mycorrhizal vs. non-mycorrhizal roots. In the same experimental system, the application of the cytokinin 6-benzylaminopurine to non-mycorrhizal roots resulted in a similar increase in Msenod40 expression. A significantly higher incidence of AM fungal colonization (vesicles per cm of root) in M. sativa cv. Gilboa roots was observed following application of luteolin-induced Sinorhizobium meliloti exudates as compared to plants amended with non-induced exudates. Moreover, overexpression of the Mtenod40 gene in Medicago truncatula plants under the control of a 35S constitutive promoter resulted in significantly enhanced colonization as compared to non-transgenic plants. The fact that enod40 gene expression was increased both in a legume and in a non-legume during colonization by the AM fungus, and that colonization levels were higher in enod40 -overexpressing transgenic plants strongly suggests that this gene is involved in the establishment of the fungal partner in its host.