In vitro mass propagation was achieved for four diverse genotypes of the wild carnation Dianthus petraeus, originating from four different geographical areas in Greece. Adventitious shoot proliferation was induced on Murashige and Skoog (MS) medium supplemented with 0.5, 1, and 2 mg l^-1 of TDZ in combination with 0, 0.1 and 0.5 mg l^-1 of NAA, using explant material from in vitro maintained stock microshoots. Hyperhydricity of the proliferated microshoots was observed and overcome by transferring the hyperhydrous microshoots to full-strength MS medium without growth regulators for four weeks prior to in vivo acclimatization. The effect of plant growth regulators was investigated and the mean number of adventitious shoots was recorded throughout all the developmental stages of micropropagation. The supplement of 0.1 mg l^-1 NAA in the growth medium is recommended for all D. petraeus genotypes but treatments with different amounts of TDZ produced variable amounts of adventitious shoots (~17-24) per explant, depending on the genotype. A significant number of proliferated shoots was observed after the exclusion of TDZ and NAA during the hyperhydricity reduction treatment and a well developed root system formed using either NAA or IBA. The regenerated plantlets with well-developed root and shoot system were successfully acclimatized in greenhouse and exterior nursery conditions.