Effect of inoculation and co-culture condition in hairy root induction in chicory (Cichorium intybus L.) by rhizogenes Agrobacterium

Background and Aim: Chicory (Cichorium intybus L.) is a member of Asteraceae and an important medicinal plants, contain a number of valuable medicinal compounds include chicoric asid, phenolic, inulin, escoline, coumarin and flavonoids. Hairy root induction by Agrobacterium rhizogenes are an effective method for production of secondary metabolites, because hairy roots are genetically and biologically stable and they are able to produce metabolite without need to hormone.Methods: In this study, through first experiment, three explants (leaf, hypocotyl and petiole) and different co-culture times were investigated. In second experiment inoculation time and also co-culture medium were examined to establish an efficient transformation system for chicory. Molecular confirmation of transgenic hairy roots was done with PCR using gene-specific primers for rolB gene and also growth rate of hairy root lines obtained from different explants were investigated.Results: The results show that maximum chicory hairy roots induction (۸۰ percent) and root number (۸.۷۱ roots per explant) and root branch (۹.۱۶ per explant) was achieved using leaf explants in combination with ۷۲ hours co-culture. In second experiment, Maximum hairy root induction percentage (۹۵.۶۵ %) and total flavonoids contents (۳۱.۲۸ mg g-۱ DW) obtained by using macro elements-free MS medium and ۱۵ min inoculation.Conclusion: Anthocyanin content was higher (۱۳.۵۵ mg g-۱ DW) in MS medium treatment, and Maximum Phenolic content (۴.۷۵ and ۴.۴۲ mg g-۱ DW respectively) obtained by using macro elements-free MS medium and MS medium.

Keywords:

Explant , Co-culture , Inoculation , Phenolic , RolB gene