Abstract:Trititrigia is the intergeneric hybrid which is from the hybridization between Triticum durum Desf. and Elytrigia intermedium (Host) Nevski. Protoplasts of Trititrigia were isolated from the embryogenic cell suspension derived from immature inflorescence-induced calli of the hybrid F1. The first division occured 48 hr after plating in modified KM8p culture medium. The plating efficiency of protoplasts was 2% and 12.14% when they were cultured in liquid medium and agarose solidified medium, respectively. Clusters grew vigorously under these conditions. Fresh medium with decreased osmoticum was added 20–30 days after plating. When protoplast-derived calli, 2–4 mm in the size, were transferred step by step to different differentiation media, embryoids, green spots emerged and numerous plants regenerated eventually.