Skip to main content

Table 3 Media composition for the transformation and regeneration of the japonica rice varieties Curinga and Nipponbare and the indica rice variety IR-64

From: Ensifer-mediated transformation: an efficient non-Agrobacterium protocol for the genetic modification of rice

Medium code Use Composition
CHU-Ind Induction and proliferation of calli. N6 salts and vitamins (Chu 1978), 100 mg l−1 Myo-inositol, 2.5 mg l−1 2,4-d, 300 mg l−1 Casein enzymatic hydrolysate, Maltose 30 g l−1, Gelrite 3 g l−1. Autoclave and supplement l-proline 500 mg l mg l−1 and l-glutamine 500 mg l−1. pH 5.8
CHU-Ind-AS Co-cultivation Chu-Ind medium supplemented with 100 µM acetosyringone. pH 5.8.
CHU-infection Agro-Infection N6 salts and vitamins (Chu 1978), 100 mg l−1 Myo-inositol, 2,5 mg l−1 2,4-d, 1 g l−1 Casamino acids, Maltose 15 g l−1, Glucose 15 g l−1. pH: 5.2. Filter sterilize. Add sterile acetosyringone 100 µM
CHU-H20
CHU-H40
CHU-H30
Selection Chu-Ind medium supplemented with l-proline 500 mg l−1, l-glutamine 500 mg l−1, Gelrite 3 g l−1, 20 mg l−1 Hygromycin (for Curinga), 40 mg l−1 Hygromycin (for Nipponbare), 30 mg l−1 Hygromycin (for IR64), 250 mg l−1 Cefotaxime. pH 5.8
MS-H20
MS-H40
MS-H30
Shoot induction/regeneration MS salts and vitamins (Murashinge and Skoog 1962), 100 mg l−1 Myo-inositol, 1 mg l−1 NAA, 30 g l−1 Sucrose, 3 g l−1 Gelrite. Autoclave and supplement with 4 mg l−1 Kinetin, 20 mg l−1 Hygromycin (for Curinga), 40 mg l−1 Hygromycin (for Nipponbare), 30 mg l−1 Hygromycin (for IR64), and 250 mg l−1 Cefotaxime. pH 5.8
MS-R-20
MS-R-40
MS-R-30
Rooting MS salts and vitamins. 30 g l−1 Sucrose, 3 g l−1 Gelrite. Autoclave and supplement with, 20 mg l−1 Hygromycin (for Curinga), 40 mg l−1 Hygromycin (for Nipponbare), 30 mg l−1 Hygromycin (for IR64), 250 mg l−1 Cefotaxime. pH 5.8