-The genus Scorzonera L. s.l. encompasses enormous morphological variation and has created problems for taxonomists. Podospermum has been treated as either a section of a broadly defined Scorzonera or as an independent genus. In Italy, there are twelve Scorzonera species and three Podospermum species. The relationship between these species is not at all clear. To clarify these relationships, a survey, including thirteen of the fifteen taxa found in Italy, was carried out using: i) sequence analysis of ribosomal internal transcribed spacer (ITS) and external transcribed spacer (ETS) regions; ii) analysis of Amplified Fragment Length Polymorphisms (AFLPs); iii) chromosome counts. In addition, to broaden the scope of the work and allow taxonomically significant interpretation of the data to be made, sequences from a range of related species were obtained from the EMBL nucleotide sequence database. Our analyses clearly indicate that Scorzonera s.l. is a polyphyletic grouping. The three approaches used were consistent with each other although showing different levels of resolution. There was broad division, on the basis of chromosome number, either 2n = 12 or 14. This was supported by PCO analysis of AFLP fragments which indicate three groups. Finally, phylogenetic analysis of sequence data produced four to six highly supported clades that might appropriately be given independent taxonomic status.