Abstract

Plant transformation mediated by Agrobacterium tumefaciens, a soil plant pathogenic bacterium, is the most used method for the introduction of foreign genesinto plant cells and the subsequent regeneration of transgenic plants. We have standardized the tissue culture media for the regeneration and transformation with the vector LBA 4404 (pCAMBIA 2301), so that in future, this system may be exploited for the expression of antibody fragment (single chain variable fragment) in plants (plantibody). The transformed green shoots tested positive for neomycin phosphotransferase II (nptII) gene and glucuronidase (GUS) were screened, rooted on MS medium and subsequently hardened to harvest seeds. The transformation frequency of Agrobacterium (LBA 4404) with the binary vector pCAMBIA 2301 on the basis of GUS resistance was found to be 2.9%.

Key words: Agrobacterium, transformation, Nicotiana tabacum, tobacco, transformation frequency.

Abbreviation

GUS, Glucuronidase;  BAP, 6-benzylamino purine; NAA, α-naphthaleneacetic acid; MS, Murashige and Skoog; nptII, neomycin phosphotransferase II; IBA, indole- 3- butyric acid; O.D, optical density; scFv,  single chain variable fragment, YEM, yeast extract mannitol; RM, regeneration medium; CTAB, cetyl trimethylammonium bromide; LB, Luria Bertani.