Protoberberine alkoloids from the hairy root cultures of Tinospora cordifolia transformed with Agrobacterium rhizogens

Recently the production of secondary metabolites using plant cells has become the subject of extended research. These secondary metabolites are a high value and low yield compounds. Evaluation of its medicinal value demands considerable attention. “Hairy“ root culture technology represents an advantageous method for the downstream processing of such important bioactive components. Thus, in the present study hairy roots of Tinospora cordifolia were induced from the shoot cultures by transformation with Agrobacterium rhizogenes strain 2402 on a solid YMB medium. Roots were subcultured on liquid MS medium containing B₅ vitamins and 3% sucrose, devoid of any plant growth hormone. Optimization of various growth parameters like light, precursor, and elicitor treatment was studied for a period of 28 days. In addition, a time course study was also carried out to understand a basic growth pattern and occurrence of alkaloids in the transformed roots. Time course analysis revealed that, berberine production was maximum on the 21^st day. However, it was observed that a higher amount of berberine (0.034%) was produced in the cultures treated with 500mg/L of L-Tyrosine as precursor, than the control. Jasmonate elicitation was found best at the concentration of 200µmoles/mL (0.047%). Moreover the berberine content in hairy roots was comparable to that produced by the roots of parent plant. HPLC and HPTLC results show the presence of jatrorrhizhine, in trace amounts. Thus it can be concluded that, the hairy root cultures form a promising source for the production of berberine and related compounds.

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