Genomic evidence for a-α heterothallic and α-α unisexual mating and recombination in an environmental Cryptococcus deneoformans population

Cryptococcus deneoformans is a human fungal pathogen capable of both a-α and α-α mating and sexual reproduction in laboratory settings. However, the extent of a-α and α-α sexual reproductions in natural populations remain unexplored. Here we analyzed the whole-genome sequences of 24 environmental strains of C. deneoformans from western Saudi Arabia, including one MATa and 23 MATα isolates, with 15 MATα isolates belonging to multi-locus sequence type ST160 as defined by their combined DNA sequences at seven loci. To identify signatures for a-α and α-α reproduction, three samples were analyzed: total, MATα, and ST160. For each subpopulation, single nucleotide polymorphisms (SNPs) were identified for both the nuclear and mitochondrial genomes and subjected to four-gamete tests. In the total population and the MATα subpopulation, variable proportions of SNP pairs within as well as between the nuclear and the mitochondrial genomes showed evidence for recombination. Though no mitogenome SNPs were found among the 15 strains of ST160, the nuclear genome showed clear evidence for recombination, including among SNPs within the MATα mating type locus. In addition, the nuclear genome SNP pairs located further apart on the same chromosome showed a greater frequency of recombination in all three sample types. In contrast, mitogenome SNPs involved in recombination were mainly located in two large genomic regions. Together, these results provide robust evidence for both a-α and α-α sexual reproduction within this environmental population of C. deneoformans.