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Fig. 2 | BMC Biology

Fig. 2

From: The broad use of the Pm8 resistance gene in wheat resulted in hypermutation of the AvrPm8 gene in the powdery mildew pathogen

Fig. 2

Functional validation of AvrPm8 in Nicotiana benthamiana. a, b Agrobacterium-mediated transient expression in Nicotiana benthamiana of BgISR7-10067-FLAG (AvrPm8) or BgISR7-10067_F43Y-FLAG (AvrPm8_F43Y) co-infiltrated with (top) or without (bottom) Pm8-HA. Leaves were imaged at 5 days post inoculation using a normal camera (a) or using a Fusion FX imager system (b). Co-infiltrations were performed with a ratio of 4 (effector): 1 (Pm8-HA). Co-expression of BgISR7-10067 with Pm8 resulted in hypersensitive cell-death (HR) in four independent experiments with a total of n = 16 leaves. Co-expression of BgISR7-10067_F43Y with Pm8 or expression of any component alone did not result in HR (n = 16). c Quantification of HR intensity in co-expression assay depicted in b. Individual datapoints are color coded based on four independent experiments with n = 4 leaves per experiment (total n = 16). d, e Detection of N. benthamiana expressed AvrPm8-FLAG and AvrPm8_F43Y-FLAG (d) or Pm8-HA (e) by anti-FLAG or anti-HA western blotting (top panel) or total protein Ponceau S staining (bottom panel). Black arrows indicate positions of protein size markers

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