The regeneration of natural tetraploid T. pratense, originated from Erzurum-Turkey, is reported in this study. This plant has low seed setting and hard seed problems due to polyploidy. Hypocotyl, cotyledon, apical meristems, epicotyl and young primary leaves were inoculated on MS and PC-L2 media containing different concentrations of BAP and NAA as growth regulators. The best shoot formation has been observed on explants initiated from apical meristem placed on PC-L2 medium that includes 2 mg dm(-3) BAP and 1 mg dm(-3) NAA. 94.4% of the shoots originated from calli were rooted on PC-L2 medium with 1 mg dm(-3) NAA. In vitro organogenesis has been accomplished in the natural tetraploid T. pratense regenerated plants successively transferred to the field.