A plasmid, pM41, has been isolated from the Spiroplasma citri strain M4 (group I-1) and characterized by restriction mapping. Using a 32P-labeled probe specific of the plasmid, we have shown by DNA-DNA hybridization that a plasmid identical to pM41 or a closely related plasmid, is present in several, but not all, S. citri strains. DNA sequences that hybridize to pM41 were also identified in three other spiroplasmas not belonging to the S. citri species. Protein patterns of several S. citri strains have been compared in order to investigate the effect of pM41 on the spiroplasma protein profiles or maps. In fact, the presence of pM41 does not appear to modify the protein pattern.