Sheltered load in fungal mating-type chromosomes revealed by fitness experiments

Sex chromosomes and mating-type chromosomes can carry large regions with suppressed recombination. As a result of a lower efficacy of selection, recessive deleterious mutations are expected to accumulate in these non-recombining regions. Multiple genomic analyses have indirectly inferred the presence of deleterious mutations in sex and mating-type chromosomes, but direct experimental evidence remains scarce. Here, we performed fitness assays in fungi with megabase–large and young non-recombining regions around the mating-type locus, using three Sordariales species, to test whether heterokaryons (diploid-like, heterozygous at the mating-type locus) exhibited a fitness advantage over homokaryons (haploid-like, with a single mating-type allele), in terms of spore germination dynamics or mycelium growth speed, under different conditions of light and temperature. We found a faster growth of heterokaryons compared to one of the homokaryons for Podospora anserina at 18 °C and for Schizothecium tetrasporum and Schizothecium tritetrasporum at 22 °C under light. These findings suggest the presence of a sheltered load, i.e., recessive deleterious mutations at the heterozygous state in or near non-recombining regions, associated to a specific mating-type allele. Genomic analyses indeed suggested that the non-recombining regions around the mating-type locus likely carry heterozygous deleterious mutations, while the rest of the genome was mostly homozygous. The difference in growth rates did not result from different numbers or densities of nuclei between homokaryons and heterokaryons. Leveraging the experimental assets of fungi, allowing cultivating separately haploid-like and diploid-like life stages, our experiments provided one of the rare direct experimental evidence of sheltered load around mating-compatibility loci.