Fig. 4.

Germline transmission of Cas9 induced mutations from T₁ to T₂ plants in barley and B. oleracea in the absence of the T-DNA construct. a Sequence alignment of T₁-181_E1 and five homozygous T₂ progeny with homozygous 1-bp deletion in HvPM19_1. b Sequence alignment from representative clones of T₁ heterozygote mutants and direct Sanger sequencing of their T₂ progeny with homozygous mutations in the absence of the T-DNA. The numbers of clones supporting T₁ mutant alleles are indicated on the right. c Sequence alignments of BolC.GA4.a Target 2 in homozygous T₁ and T-DNA free T₂ plants. Across panels the target sequences for sgRNAHvPM19-1 and sgRNABolC.GA4.a (grey) and PAM (red) are highlighted and Cas9 induced insertions and deletions are indicated by red font or red hyphens, respectively. Names of homozygous T₂ plants that lack the presence of the T-DNA construct are indicated in blue; individual homozygous plants with the same allele are shown on the same row and are labelled with a ‘p’ prefix