title: Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata.
abstract: Immature stem segments of seven different genotypes of Brassica carinata produced shoots with variable frequencies when cultured in MS medium with BAP and picloram at 0.2 mg/l each. Line 171, which produced shoots with 100% efficiency from both cut ends of the expiant, was selected for testing the amenability of this regeneration protocol for genetic transformation. A non-oncogenic Agrobacterium tumefaciens containing plasmid PCV 730, a binary vector carrying resistance genes for kanamycin and hygromycin, was used. A cocultivation period of 4 d with a bacterial concentration of approximately 2.5×10 cells/ml, followed by a recovery period of 2 d, produced transformed shoots that could be selected and rooted in the presence of kanamycin at 15 mg/l. Transformation was confirmed by neomycin phospho-transferase assay and Southern blot analysis. Seed analysis of transformed plants indicated that kanamycin resistance was inherited in the progeny.
authors.affiliation: Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India.
authors.forename: S B
authors.initials: SB
authors.lastname: Narasimhulu
authors.forename: P B
authors.initials: PB
authors.lastname: Kirti
authors.forename: T
authors.initials: T
authors.lastname: Mohapatra
authors.forename: S
authors.initials: S
authors.lastname: Prakash
authors.forename: V L
authors.initials: VL
authors.lastname: Chopra