MS medium supplemented with different concentrations of auxins 2,4-dichloro-phenoxyacetic acid (2,4-D) and α-naphthalene acetic acid (NAA) and combination with kinetin were studied to obtain a suitable protocol of callus initiation of Trigonella foenum graecum. Callus was induced from hypocotyls and cotyledons explants which were collected from the seedlings of the mentioned plant. The explants were cultured in MS medium supplemented with two auxins 2,4-D and NAA separately with different concentrations (0.0 as control, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 mg/l). Concentration of kinetin (0.5 mg/l) was used in combination with all concentrations of 2, 4-D and NAA hormones. The callus was successfully induced in all different concentrations of two mentioned auxins and combinations of different concentrations of two auxins separately with (0.5 mg/l) of kinetin. No callus formation was observed in the absence of plant growth regulators. Hypocotyl explants of T.foenum- graecum were much better in inducing callus than cotyledons explants. Combinations of NAA+Kin and 2,4-D+Kin were found to be more effective for inducing callus from hypocotyls explants compared to 2,4-D and NAA alone, at the same time callus induced by 2,4-D using cotyledons explants gave the best results compared to the other hormone. Among the different concentrations of auxin and combinations with kinetin, the highest mean of callusing index from hypocotyls segments was (3.50±0.15) with 100% of callusing in sixth week by 4.0 mg/l NAA+ 0.5mg/l Kin. In the case of callus induction from cotyledons segments the highest mean of callusing index (2.41±0.18) with 100% of callusing in sixth week was observed by 1.0 mg/l 2,4-D