Abstarct: Licorice is an important medicinal plant containing glycyrrhizic acid as a secondary mextabolite. In this study, the germination of three licorice ecotypes including Iraqi, Kashmar and Joghtay showed that the highest germination rate (80%) was obtained in Iraqi ecotype using 98% sulfuric acid. In vitro evaluation of seedling growth in solid, liquid, and hydrogel media with and without activated charcoal emphasized that the most of the morphological and physiological traits evaluated in liquid medium containing activated charcoal (2 g / l) were improved. The effects of IAA and NAA on rooting of different ecotypes and secondary mextabolite content showed that the highest fresh weight and diameter of roots were obtained in Kashmar ecotype under 1.5 and 0.5 mg / l IAA, respectively. The highest amount of glycyrrhizic acid (1.33 mg/g DW) was also produced on medium supplemented with 1 mg/l IAA in Kashmar ecotype. The effect of carbon nanotubes (0, 100 and 450 mg) and KH2PO4 (0, 2.2 and 3 g) different levels on growth of in vitro seedlings showed that increasing phosphorus and nano-carbon increased morphological and physiological traits such as Leaf, stem and root length, fresh and dry root weight and glycyrrhizic acid content in three ecotypes. The production of transgenic roots using Agrobacterium rhizogenes strain AR15834 showed the superiority of the root production in the 3-week-old of leaf explants treated with 200 micro molar acetosringone in Kashmar ecotype. In addition to these results, the highest frequency of hairy root production (60%) was also observed in 7-day-old seedling with acetosringone in Kashmar ecotype and the genetic transformation of hairy roots were validated by rolB gene PCR. baxsed on the findings of the previous steps, seedlings with transgenic roots attached to their hypocotyl were isolated and were transferred to the MS semi-solid medium containing different levels of carbon nanotubes MWCNTs (0, 100 and 450 mg) and KH2PO4 phosphorus (0, 2.2 and 3 g). The assay of Glycyrrhizic acid measured by HPLC in 14-day-old seedlings showed that Kashmar ecotype treated with 450 mg / L MWCNTs produced the highest amount of this active ingredient with an average of 16.951.33 mg/g DW.