Source: Xu, D., et al. Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice. J. Vis. Exp. (2020).
In this video, we demonstrate genetic transformation of the rice genome using an Agrobacterium tumefaciens binary vector. This bacterium facilitates the transfer of a foreign DNA plasmid using Agrobacterium virulence proteins.
1. Rice genetic transformation and plant tissue culture
Table 1: Media compositions for rice transformation. Please click here to download this table.
Figure 1: Starting materials for inducing callus. (A) Young rice inflorescences at meiosis stage. (B) Dissected young inflorescences growing on the NBD2 medium. Bar = 2 cm in (A).
Figure 2: Procedure for producing transgenic lines by Agrobacterium-mediated transformation. (A) Rice calli generated from the young inflorescences. (B) Subculture of calli. (C) Incubation of calli with A. tumefaciens. (D) Co-cultivation of calli with A. tumefaciens. (E) First selection cycle of transformed calli in the presence of Hygromycin (40 mg/L). (F) Second selection cycle of transformed calli in the presence of Hygromycin (40 mg/L). (G) First regeneration of transformed calli. (H) The second regeneration of transformed calli, noting the green shoot point in the culture plate. (I) Shoot induction culture. (J) Root induction.
The authors have nothing to disclose.