Supplementary Materials

The pan-genome effector-triggered immunity landscape of a host-pathogen interaction

Bradley Laflamme, Marcus M. Dillon, Alexandre Martel, Renan N. D. Almeida, Darrell Desveaux, David S. Guttman

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1, S4, and S7 to S15
  • Captions for figs. S2, S3, S5, and S6
  • Tables S1, S5, S7, and S8
  • Captions for tables S2, S3, S4, and S6
  • References
Figure S2
Phylogenetic trees for each family of all type III secreted effectors identified in this study. Multiple alignments were generated from the amino acid sequences of all effectors in each family using muscle and phylogenetic trees were generated with FastTree. Branches with less than 70% support were collapsed. Blue dots signify synthesized effectors from each family that elicited ETI, while red dots signify synthesized effectors that did not elicit ETI.
Figure S3
Phylogenetic trees for all families of synthesized type III secreted effectors in PsyTEC. Multiple alignments were generated from the amino acid sequences of all synthesized effectors in each family using muscle and phylogenetic trees were generated using PhyML with 1,000 bootstraps. Leaves are labelled with the cluster name of the synthesized effector and bars signify the number of effectors in the corresponding clusters across all P. syringae strains. Cluster names and labels are colored according to whether the cluster representative elicited ETI (blue) or did not elicit ETI (red).
Figure S5
Western blot summary for all synthesized type III secreted effectors from PsyTEC in PtoDC3000. HA-tagged effectors were detected using HA antibodies. Although only one representative blot is shown for each of the 529 effectors, each effector was assayed for protein expression a minimum of two times. Bands were observed for 402/529 effectors in PsyTEC (76.0%), with most effectors migrating at or around the expected kDa size indicated in parentheses beside the PsyTEC effector name. Ladder sizes are indicated to the left of each membrane. Representative images were chosen based on which exposure time best captured the overall expression on a given membrane and effectors which expressed at low levels may have only been scored as such based on longer exposure times. * = no expression detected; ** = expression detected at a significantly different size than expected; N.R. = not relevant to this study.
Figure S6
Primary screen results from representative plants sprayed with all 529 representative type III secreted effectors from PsyTEC. Each flat includes a negative control for ETI (PtoDC3000::EV) and one of two positive controls for ETI (P1: PtoDC3000::HopZ1a; P2: PtoDC3000::AvrRpm1). All plants were analyzed PIDIQ (22), which evaluates the percentage of yellow chlorotic tissue in each plant. PIDIQ values presented under each column were normalized to the negative and positive ETI controls of each flat, where the negative control was considered 100% yellow and the positive control was considered 0% yellow. ETI eliciting effectors are colored in blue and non-ETI-eliciting effectors are colored in red, based on a normalized percent yellow cut-off of 45%.
Table S2
Distribution of the members from each type III secreted effector cluster across the 494 P. syringae strains used in this study.
Table S3
Results of Western Blotting to test for in planta expression of all 529 synthesized effectors in PsyTEC.
Table S4
Disease scores for each of the 529 synthesized type III secreted effectors in this study based on PIDIQ.
Table S6
Complete nucleotide sequences of all catalytic mutant constructs used in this study.