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. 2020 Jul 8;21(9):1131–1148. doi: 10.1111/mpp.12967

Figure 6.

Figure 6

Identification of amino acids required for PaRXLR24 cell death‐triggering activity. (a) Amino acid alignment of Phytophthora sojae Avh238 (two alleles), Phytophthora parasitica pPE4, and PaRXLR24. The red star indicates the position of the 79th amino acid (histidine) in the active P6497 allele of Avh238 required for cell death‐inducing activity (Yang et al., 2017), and the corresponding 81st amino acid (isoleucine) in PaRXLR24. Black stars indicate positions of amino acids shown to affect cell death‐inducing activity in Avh238 and corresponding amino acids in PaRXLR24. (b) Agroinfiltration of PaRXLR24 wild‐type and mutants on 5‐week‐old Nicotiana benthamiana leaves. The experiment was repeated three times with consistent results. 3 × FLAG‐GFP was used as negative control. The PaRXLR24P59S and PaRXLR24A78S single (black star) mutants showed similar levels of cell death as the wild type. PaRXLRI81N single (red star), double and triple mutants showed reduced cell death compared to the wild type. Photographs were taken 7 days post‐infiltration. (c) Western blotting confirmed the stability of wild‐type (WT) and mutant PaRXLR24 proteins in total leaf extracts.