Regeneration and genetic transformation of Spanish rice cultivars using mature embryos

To establish a plant regeneration system from embryogenic callus derived from mature rice embryos, the addition of aminoacids and the effect of two macronutrient solutions MSD and N6D to the basal callus induction medium was tested in three Spanish varieties, Senia, Tebre and Bahia. Aminoacids enhanced the production of embryogenic callus in Tebre and Senia whereas in the case of Bahia, embryogenic callus, which gave rise to a high rate of differentiated shoots, was induced without aminoacids. The macronutrient solution had also to be adjusted for each variety. Pre-regeneration treatment with ABA significantly improved the regeneration rate in all media tested, independently of the media in which the embryogenic callus were induced. In a comparison of growth regulators, BA yielded more shoots than Kin in all varieties whereas the effect of the auxins NAA or IAA was dependent on the variety. Transgenic plants from the three varieties were obtained via an Agrobacterium tumefaciens transformation system, using the optimised culture media.