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A reliable protocol for transformation of Catharanthus roseus through Agrobacterium tumefaciens
Toolika Srivastava1, Sandip Das, Sudhir Kumar Sopory
1Department of Biotechnology, Jamia Hamdard, New Delhi, 110062 India.
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.
