The current research was conducted at the experimental station and tissue culture laboratory of Vegetable and Floriculture Department, Faculty of Agriculture, MansouraUniversity during the seasons of 2008 – 2010. A protocol is described for rapid and large-scale propagation of the woody aromatic and medicinal shrub lemon verbena (Aloysia triphylla) by in vitro culture of shoot tips or nodal segments from mature plants. Nodal segments were found to be more efficient than shoot tips for lemon verbena shoots regeneration on MS medium (Murashig and Skoog 1962) supplemented with BAP at 0.1 mg/L. Of the three different cytokinins ,6-benzyl aminopurine (BAP), kinetin (Kin) and thidiazrone (TDZ) evaluated as supplements to Murashige and Skoog medium (MS), BAP at concentration of 0.5 mg/L was found to be the most effective  cytokinin in inducing multiple shoots. The frequency of shoot proliferation was markedly influenced by number of subcultures, since the highest percentage of shoot multiplication (100 %) as well as large the number of shoots/ explant  (11.5 shoots) were obtained in the 4^th subculture, after which there was a declined in shoot development. Rooting was best induced (92 %) in shoots cultured on MS medium augmented with indole-3-butyric acid (IBA) at 0.5 mg/L. Mixture of soil: sand: peatmoss (1: 1: 1) was the most suitable planting substrate for hardening and its use ensured high survival frequency (100 %) of regenerated plants prior to outdoor transfer.