Soil salinity is one of the most important abiotic stresses affecting plant growth. In legumes, symbiotic nitrogen fixation in nodules is affected by salt stress, and salinity tolerance is variable among species. Genes from the High affinity K+ Transporter (HKT) family are known to play crucial roles in salt stress tolerance in different plant species. In legumes these transporters are still very poorly characterized. . Here we study the HKT transporter family from the model legume Medicago trunacatula, which is moderately tolerant to salinity. The genome of this species comprises five HKT genes, hereafter named MtHKT1;1 to MtHKT1;5. Phylogenetic analysis indicated that the MtHKT polypeptides belong to HKT subfamily 1. Three members (MtHKT1;2, MtHKT1;4 and MtHKT1;5) of the Medicago truncatula family were cloned and expressed in Xenopus oocytes. Their electrophysiological properties revealed a permeability 10 times higher for Na+ than for K+ and varying rectification properties. Expression analyses of the three MtHKT genes under different biotic and abiotic conditions suggested that MtHKT1;5 is the main transporter from this family in the root, the three genes sharing a decrease of expression in drought and salt stress conditions in non inoculated plants as well as plants inoculated with rhizobia. In the shoot, the three MtHKT would be present at similar levels independently on the applied stresses. Based on biomass and ion content analysis, the nodule appeared as the most sensitive organ to the applied salt and drought stresses. The level of expression of the three MtHKT genes was strongly decreased by both stresses in the nodule.