This chapter describes an efficient Agrobacterium-mediated genetic transformation of lentil by use of cotyledonary node explants, an optimized wounding method, and vacuum infiltration. Transformation protocol was followed by direct regeneration of transgenic shoots and micrografting of the shoots on root stocks to obtain whole-plant regeneration. The most efficient transgene expression on the axil region was obtained when the Agrobacterium KYRT1 strain was used. Gradually increasing selection pressure and repeated removal of regenerated shoots between selection steps increased the number of transgene-expressing shoots greatly. This protocol allowed 2.3 % transformation efficiency and stable transgene expression and transmission which were tracked through three generations.