Abstract

An efficient, rapid and simple Agrobacterium -mediated transformation protocol was developed for ayurvedic and pharmaceutically valuable Indian medicinal herb Bacopa monnieri (L.) using EHA 105 strain harboring the binary vector pCAMBIA 1301 containinghpt and gus genes. In vitro derived node explants were immersed in Agrobacteriumsuspension (OD600 = 0.8) for 10 min, and co-cultured on Murashige and Skoog (MS) co-cultivation medium supplemented with 0.1 mg/l NAA, 1.0 mg/l BAP and 0.1 mg/L GA₃ at 25 ± 2°C for 3 days. Putatively transformed plants were selected by the ability of node segments to produce hygromycin resistant plantlets with intermittent callus after 45 days of culture on selection medium containing 10 mg/l hygromycin and 250 mg/l cefotaxime. Gus histochemical analysis of putatively transformed plants and transgenic tissues confirmed transformation event. Polymerase chain reaction (PCR) analysis was performed to confirm the stable integration of gus transgene. A transformation frequency of 68.8% was achieved.

Key words: Brahmi, node segments, gus (β – glucuronidase), hpt (hygromycin phosphotransferase).

Abbreviation

MS, Murashige and Skoog; BAP, N₆ – Benzylaminopurine; GA₃, Gibberellic acid; NAA, α-Naphthalene acetic acid.