Fig. 2

Transgenic Arabidopsis plants resistant to BSCTV have no off-target modifications at candidate sites. a Symptoms of non-transgenic and transgenic pV86-C3, pC86-C3, and C3 Arabidopsis plants after virus inoculation. Scale bar, 3 cm. White arrows indicate infected plants with symptoms. b Virus loads detected by ddPCR in non-transgenic and transgenic pV86-C3, pC86-C3, and C3 Arabidopsis plants after virus infection (n = 3). The left panel displays concentrations of virus copies as events per microliter, as automatically determined by the software. The right panel presents the relative virus accumulation in pV86-C3, pC86-C3, and C3 plants relative to control plants (n = 3). All values are means ± S.D. ***P < 0.001, ****P < 0.0001; two-tailed Student’s t test. c Indel frequencies at ten off-target candidate sites in transgenic pV86-C3, pC86-C3, and control plants determined by deep sequencing, presented as heat maps. Genomic DNA extracted from leaves and floral structures was analyzed. Each box in the heat map represents a single sequencing experiment of an individual plant. For each site, mismatches relative to the on-target site are shown by colored boxes, and bases in the spacer sequence are numbered from 1 (most PAM-proximal) to 20 (most PAM-distal)