Fig. 3.

An example of inherited deletions in different accessions. a Schematic representation of targeted gene At3g04220 (U17), with locations of sgRNA target sites and primer binding sites for genotyping. b Deletions of At3g04220 in Col-0 reference accession, as detected by PCR with primers designated in a. Nineteen individual T1 plants were tested. Expected size of PCR product for deletion is 404 bp. c Alignment of sequence of PCR product for deletion in b (red) with wild-type sequence, which is only partially shown. d Deletions of At3g04220 in T1 plants from natural accessions, as detected by PCR; number of pooled plants in square brackets. 1: Col-0 #1 sample in b; 2: TueSB30-3 [9]; 3: Nie1-2 [7]; 4: WalHaesB4 [13]; 5: Rue3.1-31 [6]; 6: TueWa1-2, TueV-13, and HKT2.4 [5] (transformation efficiency is low for these 3 accessions). e Inheritance of At3g04220 deletion in 28 transgene-free Col-0 T2 plants, progeny of #1 T1 plant in b. Several plants show only the mutant band (top), indicating that they are homozygous for the deletion