Optimization and genetic induction of hairy roots in kelus (Kelussia odoratissima Mozaff.)

Kelus (Kelussia odoratissima Mozaff.) is one of the medicinal, spicy, aromatic and endemic herb that grows in restricted regions of Iran,. Due to the importance of its pharmaceutical compounds in traditional medicine, the excessive harvesting of kelus has put it at the risk of extinction. To protect kelus from extinction, plant tissue culture can be used to produce useful compounds and secondary metabolites. The hairy root culture is an efficient system for the production of medicinal compounds and valuable secondary metabolites in plants. For this purpose, two pathogenic strains of Agrobacterium rhizogenes (A4 and ATCC15834), two types of explants (cotyledon and hypocotyl), three levels of acetosringon (a1= 0 µmol, a2=50 µmol, and a3= 100 µmol), and three levels of glucose (b1=0, b2=0.5%, b3=1%) were applied using a factorial experiment based on a completely randomized design with three replications. The effect of such different factors on the induction and production of hairy roots in kelus was investigated. The results showed that both Agrobacterium strains were able to induce hairy roots in kelus. Also, ATCC15834 strain in MS medium with 100 µmol of acetosringon in the presence of 0.5% glucose had the highest efficiency in the production of hairy roots in cotyledon explants. The lowest amount of hairy roots was observed for A4 strain in MS medium without both acetosringon and glucose in both explants. PCR analysis was performed with rolC specific primers to ensure hairy root induction has occurred by A. rizhogenes. The results of this research showed that for the first time in Iran, it was possible to induce hairy roots in kelus explants.