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. 2019 May 23;10:2292. doi: 10.1038/s41467-019-10274-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Functional characterisation of AVRPM3^B2/C2 and AVRPM3^D3 based on natural and artificial sequence variation. a Geographical distribution and frequency of AvrPm3^b2/c2 sequence haplotypes encoding active AVRPM3^B2/C2 virulence protein variant as compared to AvrPm3^a2/f2. Underlying source data is provided in a Source Data File. b Summary of AVRPM3^B2/C2 haplovariants, only polymorphic residues are depicted. Strong recognition by PM3B and PM3C based on N. benthamiana infiltration assays are indicated with a red box, and weak recognition with an orange box. All assays are based on expression of the mature protein after removal of the signal peptide. Therefore, haplovariants E and G, and haplovariant F were not tested because the mature protein sequence is identical to variant A and variant H, respectively. c Western blot detection (upper panel) of C terminal HA epitope fusion to the mature proteins of the AVRPM3^B2/C2 haplovariants. Ponceau staining of the western blot membranes is depicted in the lower panel. Braces indicate samples where all constructs were combined on the same leaf and rotated together with AVRPM3^B2/C2-A as a reference. Uncropped Western blot images are provided in a Source Data File. d, e Functional characterisation of the impact of natural sequence diversity of AVRPM3^B2/C2 (d) and AVRPM3^D3 (e) on AVR function