Transgenic plants of mustard Brassica juncea (L.) czern and coss

Abstract

The recovery of transgenic mustard plants from the R₁ progeny of regenerated plants using cotyledon explants treated with Agrobacterium tumefaciens (C58C1, pGV3850::1103) is described. Adventitious shoots were isolated from cotyledonary callus treated with A. tumefaciens, and the shoot apices were sliced longitudinally and grown on selection medium. Regenerants were also grown on selection medium. After the second subculture, shoots with a higher ratio of green to white and/or purple were rooted. These were transplanted into soil and selfed. Seeds from R₀ generation were germinated in vitro and shoot tip and cotyledon explants were screened for kanamycin resistance. Solid green shoots were obtained from cotyledon explants on selection medium containing 20 mg/l kanamycin. These were transplanted into soil and grown to maturity. Southern blot analysis of R₁ plants showed intact integration of T-DNA into the Brassica juncea genome. This was further confirmed by seed germination and seedling growth on medium containing 200 mg/l kanamycin.