Medicinal plants are important sources of medical and many other pharmaceutical goods. The conventional propagation scheme is the main means of proliferation and takes a long period for proliferation because of poor germination and also low clonal uniformity. Rapid shoot multiplication of Gundelia tournefontii was attained from meristems on Murashige and Skoog (MS) basal medium containing 6-Benzyladenine (BAP) or kinetin, with the adding of 30 g/L sucrose. The highest number of new microshoots per explant (9.2) was attained on MS medium enhanced with BAP and 0.05 mg/L IBA. After 13 to 14 days microhoots start rooted on MS medium enhanced either Indole-3-butyric acid (IBA), Indole -3-acetic acid (IAA), or Naphthalene acetic acid (NAA) with the addition of 30 g/L sucrose. Using 100 to 300 ppm Gibbreline GA3 resulted in an increase germination percentage when soaking with GA3 for 12 hours. Gamma radiation had a significant effect on the growth of the in vitro explants. Gamma radiation had a significant effect on the germination percentage G. tournefontii seeds. The in vitro propagation plant of G. tournefontii plants could be used for the marketable clonal propagation of this important species or for future studies