Figure 4.

Inheritance and segregation of targeted mutations and male sterility generated with gRNA LTPG1‐2. (a) Crossing of the transgenic T₀ mutant line GL353‐119 with wild‐type cv. Gladius. (b) Representative examples of T₁ progeny derived from the cross shown in (a). The T₁ progeny were selfed to produce T₂ seeds. (c) Selection of DsRed‐negative (presumed non‐transgenic) T₂ seeds produced by line T1‐1. Scale bar = 1 mm. (d) Representative examples of T₂ progeny grown from the selected DsRed‐negative seeds. (e) Genotyping of the 26 DsRed/Cas9‐negative T₂ progeny. The +1 mutant allele contains an AluI restriction site that is not present in the WT allele. Cleavage at the AluI restriction site results in a 210 bp band (red arrow head).