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The obligate biotroph oomycete  Plasmopara halstedii  causes downy mildew on sunflower crop,  Helianthus annuus . The breakdown of several  Pl  resistance genes used in sunflower hybrids over the last 25 years came along with the appearance of new  Pl .  halstedii  isolates showing modified virulence profiles. In oomycetes, two classes of effector proteins, key players of pathogen virulence, are translocated into the host: RXLR and CRN effectors. We identified 54 putative CRN or RXLR effector genes from transcriptomic data and analyzed their genetic diversity in seven  Pl .  halstedii  pathotypes representative of the species variability.  Pl .  halstedii  effector genes were on average more polymorphic at both the nucleic and protein levels than random non-effector genes, suggesting a potential adaptive dynamics of pathogen virulence over the last 25 years. Twenty-two KASP (Competitive Allele Specific PCR) markers designed on polymorphic effector genes were genotyped on 35 isolates belonging to 14  Pl .  halstedii  pathotypes. Polymorphism analysis based on eight KASP markers aims at proposing a determination key suitable to classify the eight multi-isolate pathotypes into six groups. This is the first report of a molecular marker set able to discriminate  Pl .  halstedii  pathotypes based on the polymorphism of pathogenicity effectors. Compared to phenotypic tests handling living spores used until now to discriminate  Pl .  halstedii  pathotypes, this set of molecular markers constitutes a first step in faster pathotype diagnosis of  Pl .  halstedii  isolates. Hence, emerging sunflower downy mildew isolates could be more rapidly characterized and thus, assessment of plant resistance breakdown under field conditions should be improved.

Effector Polymorphisms of the Sunflower Downy Mildew Pathogen Plasmopara halstedii and Their Use to Identify Pathotypes from Field Isolates