Transformation of barrel medic (Medicago truncatula Gaertn.) by (Agrobacterium tumefaciens and regeneration via somatic embryogenesis of transgenic plants with the MtENOD12

Summary Fertile and stable transgenic plants of the model legume Medicago truncatula Gaertn. were obtained through transformation of leaf tissue with the disarmed Agrobacterium tumefaciens strain LBA4404 and in vitro regeneration via somatic embryogenesis. An optimised transformation/regeneration protocol has been established for two genotypes of the cultivar Jemalong, including a previously described highly embryogenic line (Nolan et al. 1989, Plant Cell Rep. 8: 278–281). Using this protocol, transgenic plantlets were obtained within 4–10 months following cocultivation with Agrobacterium. We have introduced into M. truncatula a chimeric fusion between the early nodulin MtENOD12 promoter and the gus (-glucuronidase) reporter gene, and shown that symbiosis-specific gene expression can be elicited in the roots of such transgenic plants following the addition of purified Rhizobium nodulation factors.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - EDTA ethylenediamine tetracetic acid - GUS gb-glucuronidase - IBA indolebutyric acid - MES 2-(N-morpholino) ethane sulfonic acid - OD optical density - X-Gluc 5-bromo-4-chloro-3-indolyl glucuronide