Fig. 1
OsIMPα1a and OsIMPα1b interact with PthXo1. a OsIMPα1a and OsIMPα1b were identified as interactors of PthXo1 in split-ubiquitin-based yeast two-hybrid (SUB-Y2H) assays of PthXo1 in combination with each of the indicated IMPs from rice. The protein‒protein interactions were detected by an X-Gal assay of colonies grown on synthetic dropout (SD)-amino acid tryptophan-leucine nutrient (SD-WL) media. b Diagram of PthXo1 structural components, including the 152-amino acid-long N-terminal sequence, which is responsible for autoactivation of the protein. The inset shows the presence and absence of colonies growing on SD-tryptophan-leucine-alanine-histidine (SD-WLAH) medium from yeast cultures that produce full-length PthXo1 protein or the PthXo1Δ152 variant lacking the first 152 amino acids. c An additional SUB-Y2H assay revealed acidic cluster (AC) motif, instead of IMPβ-binding (IBB) domain, as a recognition motif driving the interactions of OsIMPα1a and OsIMPα1b with PthXo1. In a and c, each colony represents 9 colonies investigated in 3 experimental replicates. d–f Luciferin (Luc) living fluorescence imaging (LLFI) assays performed on Nicotiana benthamiana leaves transformed with each pair of the indicated proteins. LucN and LucC represent the N-terminal and C-terminal halves of Luc fusion constructs of each tested proteins. Each leaf image represents 12 leaves of 6 plants tested in 3 experimental replicates