title: Multilocus resistance evolution to azole fungicides in fungal plant pathogen populations.
abstract: Evolution of fungicide resistance is a major threat to food production in agricultural ecosystems. Fungal pathogens rapidly evolved resistance to all classes of fungicides applied to the field. Resistance to the commonly used azole fungicides is thought to be driven mainly by mutations in a gene (CYP51) encoding a protein of the ergosterol biosynthesis pathway. However, some fungi gained azole resistance independently of CYP51 mutations and the mechanisms leading to CYP51-independent resistance are poorly understood. We used whole-genome sequencing and genome-wide association studies (GWAS) to perform an unbiased screen of azole resistance loci in Rhynchosporium commune, the causal agent of the barley scald disease. We assayed cyproconazole resistance in 120 isolates collected from nine populations worldwide. We found that mutations in highly conserved genes encoding the vacuolar cation channel YVC1, a transcription activator, and a saccharopine dehydrogenase made significant contributions to fungicide resistance. These three genes were not previously known to confer resistance in plant pathogens. However, YVC1 is involved in a conserved stress response pathway known to respond to azoles in human pathogenic fungi. We also performed GWAS to identify genetic polymorphism linked to fungal growth rates. We found that loci conferring increased fungicide resistance were negatively impacting growth rates, suggesting that fungicide resistance evolution imposed costs. Analyses of population structure showed that resistance mutations were likely introduced into local populations through gene flow. Multilocus resistance evolution to fungicides shows how pathogen populations can evolve a complex genetic architecture for an important phenotypic trait within a short time span.
authors.affiliation: Plant Pathology, Institute of Integrative Biology, ETH Zurich, Zurich, 8092, Switzerland.|School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia.
authors.forename: Norfarhan
authors.initials: N
authors.lastname: Mohd-Assaad
authors.affiliation: Plant Pathology, Institute of Integrative Biology, ETH Zurich, Zurich, 8092, Switzerland.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: McDonald
authors.affiliation: Plant Pathology, Institute of Integrative Biology, ETH Zurich, Zurich, 8092, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Croll
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D001393:Azoles; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005659:Fungicides, Industrial; D005800:Genes, Fungal; D056726:Genetic Association Studies; D005828:Genetics, Population; D001467:Hordeum; D058886:Sterol 14-Demethylase / Q000235:genetics; D050052:TRPC Cation Channels / Q000235:genetics*
chemical_list: D001393:Azoles; D005659:Fungicides, Industrial; D050052:TRPC Cation Channels; D058886:Sterol 14-Demethylase
keywords: 
  Rhynchosporium commune
  ; agricultural ecosystem; genome-wide association mapping; plant pathogens; population genomics; trade-offs