Conservation of threatened plant genetic resources is vital for food security and biodiversity maintenance. Wild species of genus Beta are invaluable germplasm which could be utilized for beet breeding programs. In this study, in vitro multiplication and genotypic responses of different species in the section Beta and Corollinae were compared in presence of plant growth regulators. Following the surface sterilization, seeds were cultivated in vitro. Cotyledonary node explants were excised from the germinated young seedlings and treated with BAP, GA3 and TDZ. The obtained results demonstrated a significant difference between the genotypes in terms of in vitro shoot proliferation efficiency when subjected to BAP and GA3. Among the applied growth regulators, effect of BAP on number of multiplied shoots was the highest. Moreover, the genotypic variation in rooting was also evident when the shoots were subjected to IAA and IBA. This study is the first report of in vitro multiplication of wild species in genus Beta that could be exploited in future germplasm maintenance, gene transformation and breeding studies.