Artemisinin, produced in very low amounts in Artemisia annua L. plants, is one of the most effective drugs in treating malaria, also has been shown to have selective toxicity towards cancer cells. Because its chemical synthesis is complicated and economically unfeasible, A. annua is currently the only source of artemisinin. According to previous studies among the Artemisia species, the highest concentration of artemisinin reported first in A. annua leaves and then in the leaves of A. dracunculus . Considering that A. annua plant is only for wet climates, cultivation and proliferation this plant Limitations faced in dry climates but tarragon cultivation is without these limitations. Thus increasing the concentration of artemisinin in tarragon plant will result in easier access and more to this drug material. Plant growth regulators (PGRs) such as Jasmonic acid and Salicylic acid stimulate growth and terpenoid biosynthesis in various aromatic plants, and play an important role in regulating the reprogramming of gene expression and in eliciting the biosynthesis of secondary metabolites in plant cells. In this study, the effect of exogenous applications of Jasmonic acid and Salicylic acid in artemisinin production and the main genes of its biosynthesis pathway such as cpr , cyp and dbr2 were investigated. For this purpose, first, to ensure uniformity of studied plants, plantlets Proliferated by tissue culture. The results of SRAP marker, confirmed the genetic stability of plants from tissue culture. Plants from tissue culture treated by 1000 micro molar Salicylic acid and 300 micro molar Jasmonic acid and sampling from them were conducted in 0, 3, 6, 12 and 24 hours after treatment. Gene expression pattern was monitored by qRT-PCR Technique. Then artemisinin content was determined by high performance liquid chromatography (HPLC) in 0, 12, 24, 48 and 72 hours after treatment. The result of the qRT-PCR experiments showed that the Jasmonic acid and Salicylic acid induced significant down-regulation of dbr2 in three hours after treatment but Jasmonic acid and salicylic acid induced significant up-regulation of cpr and cyp in six and 24 hours after treatment respectively, Previous researches showed that co-overexpressing of cyp and cpr genes could increase the artemisinin content in A.annua . Overexpressing a single gene is not sufficient to increase artemisinin accumulation, because CPR is a redox partner for CYP and helps CYP to catalyze the conversion of amorphadiene to more oxygenated products in vivo. HPLC analysis also showed that the artemisinin content was increased in 48 hours after Jasmonic acid and 72 hours after Salicylic acid treatment. According to the results of HPLC analysis, this study is also proved that co-overexpressing of cyp and cpr genes could increase the artemisinin content in tarragon. Therefore, the present study demonstrated that reduction of dbr2 expression gene had not negative effect on artemisinin biosynthesis since the artemisinic aldehyde is the last common intermediate of the two pathways leading to artemisinin or arteannuin B, respectively by DBR2 or CYP enzyme. Therefore, reducing in content of each them leads to other enzyme Produce artemisinin from one of the two pathways. It can be concluded that phyto-hormone treatment can increase some of genes involved in artemisinin biosynthesis pathway and consequently the production of artemisinin. Keywords: Artemisinin, Salicylic acid, Jasmonic acid, HPLC, qRT-PCR