Fig. 3

The AvrPm8 homologous gene in B.g. secalis is not recognized by Pm8. a Protein sequence alignment of AvrPm8 and its B.g. secalis homolog AvrPm8_Bgs. Predicted signal peptide (gray), F43Y substitution (red) and additional polymorphic residues (yellow) are highlighted. b Agrobacterium-mediated transient co-expression of AvrPm8-FLAG, AvrPm8_F43Y-FLAG, and AvrPm8_Bgs-FLAG with Pm8-HA. Leaves were imaged at 5 days post inoculation using a Fusion FX imager system (top panel). Co-infiltrations were performed with a ratio of 4 (effector): 1 (Pm8-HA). Bottom panel: quantification of HR intensity in co-expression assay depicted in top panel. Individual datapoints are color coded based on four independent experiments with n = 4 leaves per experiment (total n = 16). c Detection N. benthamiana expressed AvrPm8-FLAG, AvrPm8_F43Y-FLAG, and AvrPm8_Bgs-FLAG by anti-FLAG western blotting (top panel) or total protein Ponceau S staining as a loading control (bottom panel). Black arrows indicate positions of protein size markers. d Virulence phenotype of five B.g. secalis isolates on rye cultivars “Petkus” and “Lo7” both carrying a Pm8 gene, rye cultivar “Insave” carrying Pm17 and ‘Matador’ as a susceptible control. Pictures were taken 10 days after infection