Rosmarinus officinalis L. belongs to the Lamiaceae family and is cultivated worldwide due to its diverse uses as an ornamental evergreen shrub in gardening, as well as a food seasoning and a natural medicine. The present research was conducted to study the morphological traits of seven wild-grown rosemary populations in Greece, as well as the propagation of two selected populations with the most desirable ornamental characteristics, by the use of shoot cuttings and in vitro cultures. From the study of the morphological traits of the seven populations, those with interesting features were grown in the areas of Amaliada and Piperia and, thus, were chosen for studying their propagation. Rooting of the shoot cuttings was carried out during the four seasons of the year, using potassium salt Indole-3-butyric acid (K-IBA) at concentrations of 0.5 and 1 g·L⁻¹, in various substrates under the intermittent mist or fog system. It was found that the shoot cuttings rooted easily in all four seasons, but there were better results for the population of ‘Piperia’ in autumn, with 1 g·L⁻¹ K-IBA (80%), and ‘Amaliada’ in spring, with 0.5 g·L⁻¹ K-IBA (82.5%), while higher rooting percentages were achieved in the fog system, on a substrate consisting of perlite and peat, in a ratio of 2:1 (85%). For the in vitro cultures, shoot tips excised from the two selected populations were successfully disinfested by pre-soaking in an antioxidant solution and then, by sterilizing them in 0.6% (w/v) NaOCl, followed by transferring them onto a Murashige and Skoog (MS) nutrient medium. ‘Amaliada’ cultures proved to be the most productive population (2.1 shoots per explant), with the highest shoot formation frequency (91.6%), when cultivated on the MS nutrient medium without plant growth regulators. For ‘Piperia’ cultures, the highest shoot formation frequency (66.6%) was achieved on the MS nutrient medium supplemented with 0.25 or 0.5 mg·L⁻¹ 6-Benzyloaminopurine (BAP) and 0.1 mg·L⁻¹ Indole-3-acetic acid (IAA). Spontaneous root formation frequency was noticed on the MS nutrient medium, containing 0.5 mg·L⁻¹ BAP and 0.1 mg·L⁻¹ IAA, for both ‘Amaliada’ and ‘Piperia’ cultures (50% and 41.6%, respectively) in a single stage, with root lengths of 7.1 and 5.3 cm, respectively. Rosemary plantlets, with roots formed in vitro after transplanting them in soilless substrate, were acclimatized adequately in the greenhouse environment (~70%).