BUSCADOR RECOLECTA

The following Supporting Information is available for this article: Fig. S1 Yeast two-hybrid screening of a cDNA normalized tomato library against the CP of PepMV and identification of SlGSTU38 as a possible CP-interacting protein. Fig. S2 Phylogenetic relationship of SlGSTU38 and its homologs. Fig. S3 Mock-inoculated and PepMV-infected WT and gstu38 Micro-Tom plants at 14-d postinoculation. Fig. S4 Subcellular localization and colocalization of CP and SlGSTU38. Fig. S5 RNA-seq data validation. Fig. S6 PepMV or SlGSTU38 knockout do not induce lipid peroxidation in Micro-Tom plants. Methods S1 Tomato cDNA library and yeast two-hybrid screening. Table S1 Primer sequences. Table S2 Construct summary. Table S3 Blastp output using the SlGSTU38 protein sequence as query in the Sol Genomics Network database. Table S4 RNA-seq results: number of expressed genes per treatment and in all treatments together. Table S5 RNA-seq results: genes exclusively expressed in mock-inoculated gstu38 (gstu38_M), mock-inoculated wild-type (WT_M), and PepMV-inoculated wild-type plants (WT_PepMV). Table S6 RNA-seq results: number of differentially expressed genes (DEGs) for all the possible pairwise comparisons between treatments. Table S7 Differentially expressed genes (DEGs) comparing mock-inoculated gstu38 with mock-inoculated wild-type (WT) plants., Peer reviewed

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The consensus sequence logos were built using the online tool WebLogo 3 [49]. The reference sequences (RefSeq) were obtained from the NCBI database, and the number of RefSeq aligned per family is indicated. The conserved invariant amino acids arginine (R) and aspartic acid (D) are highlighted within yellow boxes, and the position of the sulfur-contain amino acid (cysteine or C, and methionine or M) is highlighted in a red box, Peer reviewed

The upper part of the panel shows the genomic organization of PepMV. RdRp: RNA dependent RNA polymerase gene; TGBs: Triple gene block protein-encoding genes; CP: coat protein gene. The lower part of the panel shows the alignment of the CP region including the Cys127 codon after genotyping a PepMV-infected plant after one passage (#9), and the 8 PepCPC127S-infected plants after one passage (#1–8). The Cys127 codon is framed with a red rectangle. The Cys127 replacement was stable in all the individual passages, Peer reviewed

Western blot of a protein extract from N. benthamiana leaves expressing PepCPWT. Protein extract aliquots were oxidized with CuCl2 (lanes 1 and 2), reduced with dithiothreitol (lanes 3 and 4), or not pre-treated with any of them (lanes 5–6). Protein extracts loaded in lanes 2, 4 and 6 were incubated with methyl-polyethylene glycol-maleimide (MM(PEG)24) as described by Pant et al. (2021). Reduced cysteines are alkylated with MM(PEG)24 and one conjugation increases the protein mass by 1.24 kDa. A double band was observed in lanes 2 and 4, indicating partial CP oxidation, reduction or alkylation in the controls. The membrane was stripped and re-probed with an anti-Actin antibody as the loading control, Peer reviewed

Western blot of a protein extract from N. benthamiana leaves expressing PepCPWT. Protein extract aliquots were oxidized with CuCl2 (lanes 1 and 2), reduced with dithiothreitol (lanes 3 and 4), or not pre-treated with any of them (lanes 5–6). Protein extracts loaded in lanes 2, 4 and 6 were incubated with methyl-polyethylene glycol-maleimide (MM(PEG)24) as described by Pant et al. (2021). Reduced cysteines are alkylated with MM(PEG)24 and one conjugation increases the protein mass by 1.24 kDa. A double band was observed in lanes 2 and 4, indicating partial CP oxidation, reduction or alkylation in the controls. The membrane was stripped and re-probed with an anti-Actin antibody as the loading control, Peer reviewed

Coomassie-stained SDS-PAGE gel of protein extractions from Nicotiana benthamiana leaves expressing the CP wild type (CP) or the CP mutant CPC127S. EV: pJL-TRBO empty vector; M: Marker. CP and CPC127S are expressed at similar levels, Peer reviewed