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Manipulation of root biomass and biosynthetic potential of Glycyrrhiza glabra L. plants by Agrobacterium rhizogenes mediated transformation

Abstract:

Agrobacterium rhizogenes is gramnegative soil bacteria and the causal agent for the development of the hairy root disease in plants. Hairy roots cultures have been proven as being a natural source for secondary metabolites that are normally biosynthesized in roots of differentiated plants. Here we established an experimental condition for obtaining hairy roots culture of medicinal plant Glycyrrhiza glabra as effect of integration and expression of rolB gene from Ri plasmid of A.rhizogenes. Two Agrobacterium strains, LBR56 and LBA9402 were tested for hairy roots induction but only LBR56 strain show an increase in transformation efficiency relative to wildtype. This high productive root biomass was induced from in vitro old roots and internodes segments by direct infection with A. rhizogenes LBR56 strain. Several independent hairy roots clones were obtained and maintained in vitro and further evaluated for growth, morphology, integration and expression of Ri TDNA genes. The PCR analysis with specific primers for the rol B gene and nptII marker gene confirmed the integration of TDNA fragment of Ri plasmid from A.rhizogenes into the genome of G.glabra transgencic plants. RTPCR analysis confirmed the expression of hairy roots inducible gene. The glycyrrhizic acid (GA) yield has been monitored in the hairy roots biomass by UV spectral analysis. © 2008 Bucharest University.