Fig. 4

Phylogenetic tree of C. minus Ecp32 family members across eight isolates and three morphotyes of C. minus. Neighbour-joining tree obtained from the amino acid sequence alignment of Ecp32 proteins using Geneious Software v9.1.8. Bootstrap values are shown at the nodes as percentages. The scale bar represents 0.2 substitutions per site. The number following each morphotype ID (NZFS) refers to the protein number from the annotated genomes. The asterisk (*) indicates that the protein was incorrectly predicted, and the corrected protein sequence was used. The sequences encoded by predicted pseudogenes were excluded from the analysis. The black dot refers to the proteins that were expressed in N. benthamiana and N. tabacum using an Agrobacterium tumefaciens-mediated transient expression assay to assess their ability to elicit cell death. Cms: C. minus morphotype ‘simile’; Cmv: C. minus morphotype ‘verum’; Cmn: C. minus morphotype ‘novus’. Proteins were colour-coded according to their clustering in the phylogeny