Vacuum Infiltration Based Agrobacterium Mediated Gene Transfer to Lentil (Lens Culinaris M.) Tissues

ABSTRACT

A highly efficient Agrobacterium-mediated transformation system has been developed for lentils (Lens culinaris M.) using vacuum infiltration of cotyledonary nodes and nodal segments with Agrobacterium suspension. This procedure exhibits distinct advantages over those previously reported in lentils, in that it uses vacuum infiltration enhancive force to introduce Agrobacterium suspension into the highly regenerable cotyledonary node meristems or nodal segments, which in turn rapidly produce transgenic shoots without an intermediate callus phase. The efficiency of the system has been investigated using GUS histochemical assays and PCR amplification which were evidencing integration of the transgenes and superiority of this system over conventional Agrobacterium mediated procedures that has been so far applied for lentil transformation.