show Abstracthide AbstractOur study shows that the most recent PacBio chemistries allow rapid, easy and affordable genome assembly, even of famously challenging regions such as highly repetitive non-recombining chromosomes. This breakthrough technology will now allow studying the dynamics and evolutionary role of genome structures with unprecedented resolution and power. Our study further allows broadening our understanding of evolution of non-recombining chromosomes, in particular by contrasting the evolution of fungal mating-type chromosomes and sex chromosomes.