Fig. 2: Sclerotinia sclerotiorum SsPG1 is critical for virulence and also interacts with AtPGIP1.

a Disease symptoms and disease quantification (lesion area and relative S. sclerotiorum biomass) on pea leaves caused by WMA1, two SsPG1 mutants KoSsPG1-1 and KoSsPG1-3, and the complement strain ComSsPG1-1. Representative leaves were photographed at 2 dpi. Data represent mean ± s.d. of n = 3 biological replicates. * and *** indicate significant difference from the wildtype WMA1 at p < 0.05 and 0.001, respectively, in two-tailed t-test. The experiment was performed three times independently with similar results obtained. b Bimolecular fluorescence complementation (BiFC) assay showed that SsPG1 as well as SsPINE1 interacts with AtPGIP1. N. benthamiana leaves were agroinfiltrated with a mixture of Agrobacterium tumefaciens strains harboring constructs AtPGIP1-nRFP and SsPG1-cRFP (top panel), AtPGIP1-nRFP and SsPINE1-cRFP (middle panel), and the negative control AtPGIP1-nRFP and cRFP (lower panel). The RFP fluorescence was monitored at 2 days post-agroinfiltration using confocal laser scanning microscope. Representative images are shown. The experiment was performed three times with similar results obtained. c Co-immunoprecipitation (Co-IP) assay confirmed that both SsPG1 and SsPINE1 were associated with AtPGIP1 in planta. Co-IP constructs indicated on the top were transiently expressed in N. benthamiana leaves. Immunoprecipitations were performed with anti-GFP agarose (GFP IP) or anti-Flag agarose (Flag IP). Western blot with anti-Flag antibody showed that AtPGIP1 was coprecipitated with SsPINE1 and also SsPG1 and Western blot using anti-GFP antibody showed that SsPG1 and SsPINE1 were coprecipitated with AtPGIP1. Representative images are shown. The experiments were performed three times independently with similar results obtained. Source data are provided as a Source data file.