Genome editing in the yeast Nakaseomyces delphensis and description of its complete sexual cycle

Abstract

The environmental yeast Nakaseomyces delphensis is, phylogenetically, the closest known species to Candida glabrata, a major fungal pathogen of humans. C. glabrata is haploid and described as asexual, while N. delphensis is also haploid, but has been described as competent for mating and meiosis. Both genomes contain homologues of all the genes necessary for sexual reproduction and also the genes for Ho‐dependent mating‐type switching, like Saccharomyces cerevisiae. We first report the construction of genetically engineered strains of N. delphensis, including by CRISPR‐Cas 9 gene editing. We also report the description of the sexual cycle of N. delphensis. We show that it undergoes Ho‐dependent mating‐type switching in culture and that deletion of the HO gene prevents such switching and allows maintenance of stable, separate, MATa and MATalpha haploid strains. Rare, genetically selected diploids can be obtained through mating of haploid strains, mutated or not for the HO gene. In contrast to HO/HO diploids, which behave as expected, Δho/Δho diploids exhibit unusual profiles in flow cytometry. Both types of diploids can produce recombined haploid cells, which grow like the original haploid‐type strain. Our experiments thus allow the genetic manipulation of N. delphensis and the reconstruction, in the laboratory, of its entire life cycle.

Into the wild—Exploring the life cycles of yeasts

“…The collection of papers in this Special Issue include basic science studies focusing on features of the life cycles of yeasts. A paper by Zhou Li, Boisnard, Enache‐Angoulvant, and Fairhead (2020) compared the life cycle of the wild yeast Nakaseomyces delphensis with that of the laboratory yeast S. cerevisiae . Surprisingly, the authors found that, unlike S. cerevisiae , which is largely diploid, N. delphensis exists predominantly in the haploid state.…”