<i>Agrobacterium rhizogenes</i>-mediated hairy root production in tea leaves [<i>Camellia sinensis </i>(L.) O. Kuntze]

Abstract

430-434Leaves of tea [Camellia sinensis (L.) O. Kuntze] plants were transformed with Agrobacterium rhizogenes strain (MTCC 532) and hairy roots were induced. Among the different concentrations of acetosyringeone tested, 300 M/L was found to enhance the transformation frequency upto 70%. Murashige and Skoog (MS) medium supplemented with 30 g/L maltose and indole-3-acetic acid (IAA) at 5 mg/L was found suitable for hairy-root culture and accumulation of phenolic compounds. Confirmatory studies were carried out by PCR analysis using rol C gene primer for transformation. Amplification of the specific gene was noted in the transformant at 540 bp. High performance liquid chromatography (HPLC) analysis confirmed the higher levels of catechins and their fractions in roots arising from the transformed tissues. Catechin fractions, viz., EC, ECG, EGC and EGCG were detected both in untransformed leaves and hairy roots produced by Agrobacterium infected cells