Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Abstract

Agrobacterium mediated transformation has been widely used for research in plant molecular biology and for genetic improvement of crops exploiting its tremendous ability to transfer foreign DNA to plants. In this study, the transformation efficiency of five Agrobacterium tumefaciens strains GV2260, LBA4404, AGL1, EHA105, and C58C1 was evaluated in Nicotiana tabacum L. cultivar Samsun. The Agrobacterium strains contained the recombinant binary vector pBin19 harboring beta-glucuronidase uidA gene under 35S promoter. Neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg L-1 kanamycin. The expression of uidA gene in regenerated T0 plants was firstly analyzed by GUS histochemical analyses and later on confirmation of presence of the nptII and uidA genes in regenerated plants was determined by PCR. The highest transformation rate (20%) was obtained with the Agrobacterium strain LBA4404, followed by EHA105, GV2260, C58C1 and AGL1. The higher transformation efficiency achieved in our studies make LBA4404 Agrobacterium strain optimal for functional genomics and biotechnological applications in tobacco plants.