Fig. 6
From: High-frequency, precise modification of the tomato genome
Transmission of the targeted insertion to the next generation. a Purple coloration is visible in the embryos within the seeds. b Scheme of the multiplexed PCR used to detect both WT and GT events in progeny of GT lines. Primers TC097F, ZY010F and TC210R (marked by arrows) were used in a single reaction. c A sample gel picture with products from PCR analysis of 30 T1 seedlings (gel pictures from PCR analysis of all 175 screened seedlings are provided in Fig. S12 in Additional file 1). All three possible genotypes were detected. Green arrow marks the WT products, the purple arrow the GT products, and red arrow the 1.0-kb band in the DNA ladder. The phenotype of each seedling is marked by P (purple) or G (green). M 2-Log DNA ladder (New England Biolabs), NT no template control. d–f Pictures of three of each homozygous WT (d) and heterozygous (e) and homozygous (f) GT T1 plants. The homozygous GT plants have reduced growth due to excessive accumulation of anthocyanins. Scale bars = 1 cm