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Secondary constrictions were determined in one chromosome pair in the complements of M. infesta, M. macrocarpa, M. italica (subgenus Micromeliotus), and M. alba and M. officinalis (subgenus Meliotus). The karyotypes of M. infesta and M. macrocarpa were found to be similar. Chromosomes of M. italica were larger than the chromosomes of the other four Melilotus species. The chromosome size in M. italica suggests the presence of large chromosomes in an ancestral or pro-Melilotus prototype. The chromosomes with satellites of M. infesta and M. macrocarpa appear to differ in morphology from the satellite chromosomes of M. alba and M. officinalis by a paracentric inversion. The morphology of the satellite chromosomes in M. italica is thought to represent a more primitive type than in the other Meliotus species studied.