Supplementary Figure 11: CRISPR/Cas9-induced mutations in PL in transgenic tomato lines and analysis of pericarp texture and ripening properties
From: Genetic improvement of tomato by targeted control of fruit softening
a, Target site for guide RNA (Cas9/sgRNA) used to edit the PL coding sequence by the protocol described in Nekrasov et al 45, thirteen transgenic plants harbouring the Cas9 gene were analysed by PCR and sequencing to identify mutations and the lines were then self –pollinated and homozygous mutants were identified by sequencing. Two CRISPR lines, PLC5 and PLC11, from independent transgenic events, yielded the same point mutation and the sequence is shown in comparison to wild type. The single bp insertion results in a stop codon in the PL coding sequence. b, Analysis of fruit from lines PLC5 and PLC11 harbouring the PL mutation and the azygous wild type control (WT) revealed fruits with significantly (Ftest; P≤0.04) firmer texture compared to the control as denoted by *. However there were no significant (Ftest; P> 0.05) differences in fruit colour or soluble solids content between PL and WT fruits. Error bars are s.e.m. Dots represent individual fruits. Fruit numbers were WT n=3, PLC11 n= 3, PLC 5 n=13.
