Novel secreted effectors conserved among smut fungi contribute to the virulence of Ustilago maydis .

Fungal pathogens deploy a set of molecules (proteins, specialized metabolites, and sRNA), so called effectors, to aid the infection process. In comparison to other plant pathogens, smut fungi have small genomes and secretomes of 20 Mb and around 500 proteins, respectively.. Previous comparative genomic studies have shown that many secreted effector proteins without known domains i.e., novel, are conserved only in the Ustilaginaceae family. By analyzing the secretome of 11 species within Ustilaginaceae, we identified 53 core homologous groups commonly present in this lineage. By collecting existing mutants and generating additional ones, we gathered 44 Ustilago maydis strains lacking single core effectors as well as 9 strains contain-ing multiple deletions of core effector gene families. Pathogenicity assays revealed that 20 of these 53 mutant strains were affected in virulence. Among the 33 mutants that had no obvious phenotypic changes, 13 carried additional, sequence-divergent, structurally similar paralogs. We report a virulence contribution of seven previously uncharacterized single core effectors and of one effector family. Our results help to prioritize effectors for understanding U. maydis virulence and provide genetic re-sources for further characterization.