Three allelic nuclear mutants affected in the recombination of mtDNA have been characterized in Saccharomyces cerevisiae and assigned to the PIF locus. In the mutants, the general recombination measured by the recombination frequency between linked or unlinked alleles is normal. However, the pif mutations prevent the integration into the rho+ genome of the markers (oli1, oli2, diu1, ery, oxi1, oxi2) of those rho- genomes that have tandemly arrayed repeat units. Therefore, these rho- genomes characterize a PIF-dependent recombination system. The pif mutations have also revealed the existence of a PIF-independent recombination system used by those rho- genomes that have an inverted organization of their repeat units. The markers of such palindromic rho- genomes exhibit high integration frequency into the rho+ genome even in the presence of the pif mutation. In addition, the pif mutations greatly increase suppressiveness in crosses between pif rho+ strains and PIF-dependent as well as PIF-independent rho- clones. We conclude that the recombination between rho+ and rho- genomes involves at least two distinct systems that depend on the organization of the rho- genome.