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Nucleo-mitochondrial interactions, particularly those determining the primary divergence of biological species, can be studied by means of xenomitochondrial cybrids, which are cells where the original mitochondria are substituted by their counterparts from related species.  Saccharomyces cerevisiae  cybrids are prepared simply by the mating of the ρ 0  strain with impaired karyogamy and germinating spores from other  Saccharomyces  species and fall into three categories. Cybrids with compatible mitochondrial DNA (mtDNA) from  Saccharomyces paradoxus  CBS 432 and  Saccharomyces cariocanus  CBS 7994 are metabolically and genetically similar to cybrids containing mtDNA from various  S. cerevisiae.  Cybrids with mtDNA from other  S. paradoxus  strains , S. cariocanus, Saccharomyces kudriavzevii, and Saccharomyces mikatae  require a period of adaptation to establish efficient oxidative phosphorylation. They exhibit a temperature-sensitive phenotype, slower growth rate on a non-fermentable carbon source and a long lag phase after the shift from glucose. Their decreased respiration capacity and reduced cytochrome  aa3  content is associated with the inefficient splicing of  cox1 I3β, the intron found in all  Saccharomyces  species but not in  S. cerevisiae.  The splicing defect is compensated in cybrids by nuclear gain-of-function and can be alternatively suppressed by overexpression of  MRP13  gene for mitochondrial ribosomal protein or the  MRS2, MRS3 , and  MRS4  genes involved in intron splicing.  S. cerevisiae  with  Saccharomyces bayanus  mtDNA is unable to respire and the growth on ethanol–glycerol can be restored only after mating to some  mit  −  strains. The nucleo-mitochondrial compatibility limit of  S. cerevisiae  and other  Saccharomyces  was set between  S. kudriavzevii  and  S. bayanus  at the divergence from  S. cerevisiae  about 15 MYA. The  MRS1-cox1 S. cerevisiae / S. paradoxus  cytonuclear Dobzhansky–Muller pair has a neglible impact on the separation of species since its imperfection is compensated for by gain-of-function mutation.

Post-zygotic sterility and cytonuclear compatibility limits in S. cerevisiae xenomitochondrial cybrids