title: Agrobacterium tumefaciens-mediated transgenic plant production via direct shoot bud organogenesis from pre-plasmolyzed leaf explants of Catharanthus roseus.
abstract: Production of Agrobacterium tumefaciens-mediated transgenic plants, via direct shoot bud organogenesis from leaves of Catharanthus roseus, is reported. A. tumefaciens harbouring the plasmid pBI121 with GUS gene uidA and kanamycin resistance gene nptII was used. Highest transformation efficiency of 1.4 transgenic shoots/responded explant was obtained when pre-plasmolysed leaves, pre-incubated on shoot bud induction medium for 10 days, were subjected to sonication for 30 s prior to transformation. Using a selection medium containing 50 mg kanamycin l(-1), transformants grew into micro-shoots and formed roots on a hormone-free half strength MS medium. The transgenic nature of the regenerated plants was confirmed by PCR amplification of uidA gene and GUS histochemical assay.
authors.affiliation: Department of Plant Biotechnology, Central Institute of Medicinal and Aromatic Plants, Council of Scientific and Industrial Research, Post Office CIMAP, Kukrail Picnic Spot Road, Lucknow 226015, India.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Verma
authors.forename: Ajay Kumar
authors.initials: AK
authors.lastname: Mathur
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D030008:Catharanthus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018515:Plant Leaves / Q000235:genetics; D018520:Plant Shoots / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D010957:Plasmids; D014170:Transformation, Genetic