Background: Lemon verbena (Lippia citriodora H.B.K.) is an aromatic and medicinal plant of family Verbenaceae, which cultivated in North region of Iran. Objective: Evaluation of phytochemical characters in Lippia citriodora H.B.K. plantlets affected by propagation methods (micro-propagation and stem cutting) cultivated in different soil type (peat moss and mixture soil). Methods: This study was done on the base of factorial experiment in randomized complete block design with three replications and four treatments. The treatments were propagation methods (micro-propagation and stem cutting) and soil type (peat moss and mixture soil). The volatile oil was extracted by hydro-distillation using clevenger-type apparatus and analyzed by GC and GC/MS. Results: The results indicated that the interaction effects of different soils and plantlets types had significant effect (p<0.05) on all studied parameters. The highest content of essential oil (0.79%), geranial (53.52%), neral (31.82%), limonene (11.29%), leaf dry weight (1.43 g) and SPAD value (20.85) were observed in treatment of MPP (micro-propagation plantlets cultivated in peat moss) and the lowest of that was obtained in treatment of SCM (stem cutting plantlets cultivated in mixture soil). Conclusion: The peat moss was the best bed in respect of phytochemicals and morpho-physiological traits in both types of in vitro and stem cutting propagation. The highest essential oil content was observed in treatment of micro-propagation cultivated in peat moss.