Fig. 4: Segregation analysis of BC1-F2 populations and identification of homozygous recombinant plants.
From: Demonstration of targeted crossovers in hybrid maize using CRISPR technology
a Schematic diagram of the segregation analysis of the BC1-F2 populations; b The result of the genotyping analysis of the BC1-F2 segregating population. An example of the segregation analysis of the SNP markers in 21 BC1-F2 plants from Event-2.11 is shown; c Confirmation of DNA editing pattern at the site of the targeted CO in homozygous BC1-F2 plants with Illumina sequencing. DNA was isolated from a single leaf disc sampled from each BC1-F2 plant for library construction and sequencing. Each bar on the graph represents a result derived from a single BC-F2 plant (n = 1). The proportion of edited (blue) and non-edited (red) amplicons in homozygous recombinant plants from three evaluated populations is shown on the graph. The pictures under the graph represent alignments of the sequences at the site of targeted CO with the inbred reference. gRNA1 target sequence is highlighted in yellow. The gray highlights nucleotides microhomology that could contribute to the repair of DSBs. S2d23 is an abbreviated description of the DNA editing pattern. S2 indicates the nucleotide position where the deletion starts; d23 indicates the length of the deleted nucleotides.
