Paconia species arc known for their rich chemical content and their compounds exhibit different medicinal properties. In this study, we firtly investigated the essential oil (EO) content of the root, scape, fruit and fruit bark of Paeonia daurica Andrews subsp, Macrophylla Albow (P, daurica) by using gas chromatography-mass spectrometry (GC-MS). Both solid-phase micro-extraction (SPME) and hydro-distillation (HD) were used to prepare EO extracts for each plant sample. While fifty-two compounds were identified in SPME-EO extract, sixty-eight compounds were identified in HD-EO extract and eighteen compounds were jointly found in both extract. Major constituents of the EOs were salicylaldehyde (100-65%), myrtanal (3.10-45%), palmitic acid (2.1%-40.4%), methyl salicylate (3.1%-37.5%) and myrtenal (3%-35.1%). Fruit bark had the richest identified EOs among plant parts. Furthermore, the total SPME-EO and HD-EO extracts were evaluated for their inhibition on Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV-RT) by using primer extension assay (measuring of single dATP nucleotide addition). M-MLV-RT could not add a single nucleotide to the 14-mer primer in the presence of both EO extracts and it revealed that both SPME and HD total EO extracts had strong inhibition effect on M-MLV-RT. alpha-bisabolol, beta-bisabolol and benzoic acid-ethylester (a derivate of benzoic acid) in HD-fruit extract were evaluated as potential compounds responsible for the inhibitory effect according to literature, To the best of our knowledge, this is the first detailed report about EO compositions and its inhibition effect against M-MLV-RT of the root. scape. fruit and fruit bark of P. daurica. We propose the EOs of P. daurica might have potential of therapeutic candidate for preventing viral diseases.