title: Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant.
abstract: This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%.
authors.affiliation: National Botanical Research Institute, Council of Scientific and Industrial Research, Rana Pratap Marg, Lucknow 226001, UP, India.
authors.forename: Vibha
authors.initials: V
authors.lastname: Pandey
authors.forename: Pratibha
authors.initials: P
authors.lastname: Misra
authors.forename: Pankaj
authors.initials: P
authors.lastname: Chaturvedi
authors.forename: Manoj K
authors.initials: MK
authors.lastname: Mishra
authors.forename: Prabodh K
authors.initials: PK
authors.lastname: Trivedi
authors.forename: Rakesh
authors.initials: R
authors.lastname: Tuli
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D018520:Plant Shoots / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D010946:Plants, Medicinal / Q000235:genetics*; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic; D032341:Withania / Q000235:genetics*
chemical_list: D018744:DNA, Plant