The Myrtle (Myrtus communis L.) is an evergreen shrub typical of the Mediterranean area; it is an interesting plant with multipurpose use. The ornamental use takes into account the production of green cut branches for indoor decoration and production of pot plants for gardening. In this species, there is a great variability in the natural germplasm around the Mediterranean coasts for type and size of fruit, plant architecture, leaf size and internode length. Selected genotypes have been successfully sterilized and cultured in vitro. The shoots were multiplied on MS (16) salts and vitamins, with 0.5 mg/L BA and 0.2 mg/L IAA. Clones showed variation of multiplication rate and rooting percentage. IAA or IBA at 0.5 mg/L increased the rooting percentage and noticed differences in root number and length. The sucrose concentration can affect rooting, such as light intensity during the in vitro rooting phase can modulate biomass production and chlorophyll content. The combination of these factors enhanced the frequency rate of acclimatization.