Figure 4

Four primer sets with the HRM curve for the two parents, Cowiche and Selah, on the left and the 13 seedlings on the right. HRM derivative plots, (-d/dT) fluorescence as a function of temperature. A-C contain 1 putative SNP while D. contains 2 putative SNPs. A. Primer set 131 shows no variation as expected for crossing two of the same homozygotes. Note the single curve profile in both the parents and seedlings. B. Primer set 189 shows a single curve profile in the parent panel which differentiates into a 1:2:1 (3:6:4) genotype ratio as expected for a heterozygous × heterozygous cross in the seedlings. It is represented by three different curve profiles where profile number 1 corresponds to the heterozygous parental profiles. C. Primer set 100 shows a 1:1 (8:5) genotype ratio as expected in a homozygous and heterozygous cross. The curve profiles are similar between the seedlings and the parents. D. Primer 92 shows 2 parental curve types. However, the seedlings show several distinct curve types which is not unexpected due to the presence of 2 high quality SNPs in this region.