title: A reliable protocol for transformation of Catharanthus roseus through Agrobacterium tumefaciens.
abstract: Proliferation of axillary shoot buds and multiple shoot formation in Catharanthus roseus was obtained in 96 % explants on MS medium (3 % sucrose) containing NAA + BA. 2,4-D induced callusing in both, the nodal as well as in leaf segments. Leaf-derived callus was used for transformation with Agrobacterium tumefaciens LBA4404/pBI-S1. Bacterial cell concentration, duration of co-cultivation and acetosyringone concentration influenced transformation efficiency. Under optimal co-cultivation conditions, 98 % of the explants showed GUS expression. PCR based amplification of the transformed and subsequently selected callus tissue indicated the presence of uidA, Gly I and nptII genes.
authors.affiliation: Department of Biotechnology, Jamia Hamdard, New Delhi, 110062 India.
authors.forename: Toolika
authors.initials: T
authors.lastname: Srivastava
authors.forename: Sandip
authors.initials: S
authors.lastname: Das
authors.forename: Sudhir Kumar
authors.initials: SK
authors.lastname: Sopory
authors.forename: P S
authors.initials: PS
authors.lastname: Srivastava
keywords: Agrobacterium tumefaciens; Catharanthus roseus; Co-cultivation; GUS; GlyI; Micropropagation; RAPD; Transformation