Mating-type locus rearrangements and shifts in thallism states in Citrus-associated Phyllosticta species

Currently, eight Phyllosticta species are known to be associated with several Citrus hosts, incorporating diverse lifestyles: while some of them are endophytic (P. capitalensis and P. citribraziliensis), others are pathogenic (P. citriasiana, P. citricarpa, P. citrichinaensis and P. paracitricarpa). Sexual reproduction plays a key role in the interaction between these Phyllosticta species and their Citrus hosts, especially for the spread and persistence of the pathogenic species in the environment. Given this, differences in sexual reproduction strategies could be related to the differences in lifestyles. To evaluate this hypothesis, we characterized the mating-type loci of six Citrus-associated Phyllosticta species from whole genome assemblies. Mating-type genes in the Citrus-associated Phyllosticta species are highly variable in their sequence content, but the genomic locations and organization of the mating-type loci are conserved. Phyllosticta citriasiana, P. citribraziliensis, P. citricarpa and P. paracitricarpa are heterothallic, while P. capitalensis and P. citrichinaensis are homothallic. In addition, the P. citrichinaensis MAT1-2 idiomorph occurs in a separate location from the mating-type locus. Ancestral state reconstruction suggests that homothallism is the ancestral thallism state in Phyllosticta, with a shift to heterothallism in Phyllosticta species that are pathogenic to Citrus. Moreover, the homothallic strategies of P. capitalensis and P. citrichinaensis result from independent evolutionary events, as P. capitalensis locus likely represents the ancestral state, and P. citrichinaensis homothallism has risen through a reversion in a heterothallic ancestor and underwent remodelling events. As the pathogenic species P. citriasiana, P. citricarpa and P. paracitricarpa are heterothallic and incapable of selfing, disease management practices focused in preventing the occurrence of sexual reproduction could assist in the control of Citrus Black Spot and Citrus Tan Spot diseases. This study emphasizes the importance of studying Citrus-Phyllosticta interactions under evolutionary and genomic perspectives, as these approaches can provide valuable information about the association between Phyllosticta species and their hosts, and also serve as guidance for the improvement of disease management practices.