Development of Hairy Root Culture of Andrographis Paniculata for in Vitro Adrographollide Production

Abstract

Development the hairy root culture of A. paniculata was conducted for growing the hairy roots and production of andrographollide. Different strains of Agrobacterium rhizogenes (R-1000, A4, ATCC 15834), different types of explants (cotyledons, hypocotyls, and leaves) and different infection time of A. rhizogenes (1, 2, 3 days) were tested to induce hairy roots of A. paniculata. The results indicated that the best strain, type of explants and infection time for hairy roots induction were found in strain ATCC 15834, the explants of cotyledon and the 2 days of infection, respectively. The best medium for growing the hairy roots was liquid half strength MS medium with the addition of 5.0 μM IBA. The highest amount of andrographollide was observed in the medium with the addition of 5.0 μM IBA on the week of two, as much as 0.54%. Integration of T-DNA of A. rhizogenes in hairy roots was confirmed by polymerase chain reaction (PCR) analysis with specific primer for rolA and rolC genes of the plasmid. Visualization of the PCR products on agarose gel electrophoresis showed two fragments with lengths of 248 bp and 490 bp which corresponds to rolA and rolC genes from Ri plasmids of ATCC 15834. 