Supplementary Figure 10: CRISPR/Cas9-engineered mutations in SlCLV2 result in weakly fasciated plants.

(a) Schematic illustrating two guide RNAs (sgRNAs; red arrows) targeting the SlCLV2 coding sequence. Cas9/sgRNA1/sgRNA2 were expressed from the same construct. Black arrows indicate the PCR primers used to evaluate mutation type and efficiency. (b) PCR genotyping of six T₀ CR-slclv2 plants showing the presence of indel mutations from predominantly chimeric plants, two of which carry the desired deletion. PCR for the presence of Cas9 is shown along with evaluation for fasciation. All five CR-slclv2 fasciated plants exhibited similar severity. (c) Representative primary inflorescence from an slclv2 plant (CR-slclv2-4) showing weak fasciation like fab and CR-fab mutant plants. Images of a representative flower and fruit are shown (insets). Scale bar, 1 cm. (d) Quantification and statistical comparisons of floral organ numbers from WT and CR-slclv2 flowers. Flower fasciation in CR-slclv2 plants is slightly weaker than in fab and CR-fab mutant plants (Fig. 1h and Supplementary Fig. 9). Data were collected from three plants with confirmed mutations in all sequenced alleles (e; see also Online Methods). Data are means ± s.d., n ≥ 10. A two-tailed, two-sample t test was performed, and significant differences are represented by black asterisks: **P < 0.001. (e) CR-slclv2 alleles identified by cloning and sequencing PCR products from the SlCLV2 targeted region from three fasciated T₀ plants. The CR-slclv2-3 and CR-slclv2-5 plants were chimeric, and all alleles carried indels (blue dashed lines and letters), including the desired deletion in CR-slclv2-4. One of each representative allele identified from sequencing at least eight cloned PCR products is shown. Red font highlights sgRNA target sites, and black rectangles indicate PAM sequences; a, allele.