Table 3. Identity of RNAi sequences cloned into the TRV2 vector against the N. benthamina PDS gene predicted by SGN VIGS at default parameters.

The National Center for Biotechnology Information accession number for the full-length NbPDS sequence used in this study is DQ469932.1, and N. benthamiana version 0.4.4 was selected as the database.

Name of Construct	Range on Target Sequence	Length	No. of siRNAs	Construct sequences	No. of Off-Targets	Significant Off-Targets (No. of Hits)
NbPDS (SGN VIGS tool fragment)	1236–1386	150	Not available	CCA​TAT​ATG​GTT​TGA​CAG​AAA​ACT​GAA​GAA​CAC​ATC​TGA​TAA​TCT​GCT​CTT​CAG​CAG​AAG​CCC​GTT​GCT​CAG​TGT​GTA​CGC​TGA​CAT​GTC​TGT​TAC​ATG​TAA​GGA​ATA​TTA​CAA​CCC​CAA​TCA​GTC​TAT​GTT​GGA​ATT​GGT​ATT​TGC​ACC​C	Not Available	Off-target prediction is not available in the SGN VIGS tool
NbPDS (conventionally used fragment)	855–1263	409	Not available	GGC​ACT​CAA​CTT​TAT​AAA​CCC​TGA​CGA​GCT​TTC​GAT​GCA​GTG​CAT​TTT​GAT​TGC​TTT​GAA​CAG​ATT​TCT​TCA​GGA​GAA​ACA​TGG​TTC​AAA​AAT​GGC​CTT​TTT​AGA​TGG​TAA​CCC​TCC​TGA​GAG​ACT​TTG​CAT​GCC​GAT​TGT​GGA​ACA​TAT​TGA​GTC​AAA​AGG​TGG​CCA​AGT​CAG​ACT​AAA​CTC​ACG​AAT​AAA​AAA​GAT​CGA​GCT​GAA​TGA​GGA​TGG​AAG​TGT​CAA​ATG​TTT​TAT​ACT​GAA​TAA​TGG​CAG​TAC​AAT​TAA​AGG​AGA​TGC​TTT​TGT​GTT​TGC​CAC​TCC​AGT​GGA​TAT​CTT​GAA​GCT​TCT​TTT​GCC​TGA​AGA​CTG​GAA​AGA​GAT​CCC​ATA​TTT​CCA​AAA​GTT​GGA​GAA​GCT​AGT​GGG​AGT​TCC​TGT​GAT​AAA​TGT​CCA​TAT​ATG​GTT​TGA​CAG​AAA​ACT​GAA​G	Not Available	Conventionally used fragment for VIGS