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Background  Isogamous organisms lack obvious cytological differences in the gametes of the two complementary mating types. Consequently, it is difficult to ascertain which of the two mating types are homologous when comparing related but sexual isolated strains or species. The colonial volvocalean algal genus  Gonium  consists of such isogamous organisms with heterothallic mating types designated arbitrarily as  plus  or  minus  in addition to homothallic strains. Homologous molecular markers among lineages may provide an “objective” framework to assign heterothallic mating types.    Methodology/Principal Findings  Using degenerate primers designed based on previously reported  MID  orthologs, the “master regulator” of mating types/sexes in the colonial Volvocales,  MID  homologs were identified and their presence/absence was examined in nine strains of four species of  Gonium . Only one of the two complementary mating types in each of the four heterothallic species has a  MID  homolog. In addition to heterothallic strains, a homothallic strain of  G .  multicoccum  has  MID . Molecular evolutionary analysis suggests that  MID  of this homothallic strain retains functional constraint comparable to that of the heterothallic strains.    Conclusion/Significance  We coordinated mating genotypes based on presence or absence of a  MID  homolog, respectively, in heterothallic species. This scheme should be applicable to heterothallic species of other isogamous colonial Volvocales including  Pandorina  and  Yamagishiella . Homothallism emerged polyphyletically in the colonial Volvocales, although its mechanism remains unknown. Our identification of a  MID  homolog for a homothallic strain of  G .  multicoccum  suggests a  MID -dependent mechanism is involved in the sexual developmental program of this homothallic species.

Distribution of the Sex-Determining Gene MID and Molecular Correspondence of Mating Types within the Isogamous Genus Gonium (Volvocales, Chlorophyta)