The monotypic Zhumeria majdae (Lamiaceae), an endangered medicinal and aromatic plant in the south part of Iran, has a low propagation rate in natural condition and therefore an efficient method for the in vitro-propagation is required. In the present study, the effects of source and position of the explants as well as various concentrations of 6-benzylaminopurine and kinetin on inhibition of hyperhydricity and in vitro mass propagation of the plant were investigated. The best shoot formation (1.9 ± 0.07) was obtained with a Murashige and Skoog medium fortified with 17.7 μM 6-benzylaminopurine. Regenerated shoots, elongated on the MS medium containing 2.2 μM 6-benzylaminopurine, were rooted on the different tested media, with the most abundant (68.6 ± 4.1%) and strongest roots obtained on half-strength medium without plant growth regulators. Hydrodistilled essential oils of in vitro regenerated plant were analyzed by gas chromatography–mass spectrometry and compared with the essential oil of wild plant. Thirty and twenty compounds representing 98.2% and 98.7% of the total oils were identified in the oils of wild plant and in vitro regenerated plant, respectively. The major essential oil components were linalool (31.7% and 41.7%), camphor (28.0% and 32.4%), limonene (4.6% and 8.3%), camphene (4.1% and 3.5%) and E-caryophyllene (1.0% and 2.7%) in the studied (wild plant and in vitro regenerated plant) essential oils. The in vitro regeneration system could be utilized for both conservation, largescale multiplication and production of rich linalool essential oils of Z. majdae.