Virus resistance to infection by ICMV-SG. (A) PCR analysis of genomic DNAs prepared from WT and transgenic lines. Upper panel: PCR product using P1 and P2 primers. Middle panel: PCR product using P3-1 and P4-1 primers. Lower panel: PCR product using F and R primers which are upstream and downstream flanking sequences, respectively, from the T-DNA (Transferred DNA) insertion position for line 82. Note that lines 82–4 and 82–11 were hemizygous and marker-free, and lines 82–5 to 82–10 were hemizygous but chimeric with respect to the marker gene. Lines 82–12 and 82–13 were homozygous non-marker free, and line 82–14 was a null segregant. (B) Virus symptoms in putative homozygous line 82–4, and (C) and (D): null segregant plant 82–14 (C) and WT JcMD plant (D) after vacuum infiltration of ICMV-SG infectious clones. Bar: 10 cm. (E) Quantitative PCR analysis of virus titers after virus challenge. WT plants with mock treatment was used as controls and the average control value was set as 1.