Natural cyanamide (NH 2 CN) has recently been found in three Leguminosae plants: Vicia villosa subsp. varia, Vicia cracca and Robinia pseudo-acacia. As cyanamide has long been thought to be absent in nature, its physiological role and biosynthesis are totally unknown. N]-glutamine, indicating that these nitrogenous compounds are distant precursors of cyanamide. Although the guanidino group of l-arginine (-NH-C(NH 2 )=NH) and urea (NH 2 C(=O)NH 2 ) were candidate precursors of cyanamide on the basis of structural similarity, direct incorporation of the guanidino group of l-[ 13 C 6 , 15 N 4 ]-arginine and [ 13 C, 15 N 2 ]urea into cyanamide was not observed. These results eliminated the possibility that cyanamide is biosynthesized by the addition of ammonia to an electrophilic carbon or by the conversion of the tested compounds that were structurally relevant to cyanamide.