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. 2021 Sep 20;20(1):168–182. doi: 10.1111/pbi.13704

Figure 7.

Figure 7

Sequence variations of TaCYP78A5‐2A and their associations with grain yield‐related traits. (a) Two haplotypes (Ap‐HapI and Ap‐HapII) based on the sequence variation in the promoter region of TaCYP78A5‐2A. (b) A cleaved amplified polymorphic sequence (CAPS) marker developed based on −1191 bp (C/T) with restriction endonuclease HhaI showed in (a). After enzyme digestion, the Ap‐HapI be cleaved into 170 and 140 bp, but Ap‐HapII could not be cleaved. (c) The relative activity of TaCYP78A5 promoters with haplotype Ap‐HapI and Ap‐HapII. The two haplotypes (Ap‐HapI and Ap‐HapII) of TaCYP78A5‐2A were separately cloned into the pGreenII 0800‐LUC (firefly luciferase) to generate expression vectors Ap‐HapI::LUC and Ap‐HapII::LUC, and the two constructs were separately infiltrated into the leaves of 30‐day‐old Nicotiana benthamiana, with the expression of renilla luciferase (REN) as an internal control. The LUC activity and REN activity were measured with GloMax‐multi‐luminescence reader. The ration of LUC activity to REN activity (LUC/REN) indicates the relative expression level of TaCYP78A5 Ap‐HapI or Ap‐HapII. Data are shown as the mean ± SE (n = 3). (d, e) Association of Ap‐HapI and Ap‐HapII with thousand‐grain weight (TGW) (d) and yield per plant (e) of 323 wheat accessions at 16 environmental sites. E1–E16 indicate the environments at Shunyi in 2015 under drought stressed (DS), DS + heat stress (HS), well‐watered (WW), WW + HS; Shunyi in 2016 under DS, DS + HS, WW, WW + HS; Changping in 2016 under DS, WW; Shunyi in 2017 under DS, DS + HS, WW, WW + HS; Changping in 2017 under DS and WW. (f) The nucleotide diversity (π) and Tajima’s D of TaCYP78A5‐2A promoter regions of 43 landraces and 42 cultivars. (g) Frequency changes of the two haplotypes of TaCYP78A5‐2A over decades in modern cultivars (total of 311 accessions). 14, 33, 47, 34, 60 and 123 accessions were released in pre‐1960s, 1960s, 1970s, 1980s, 1990s and post‐2000 respectively. Ap‐HapI and Ap‐HapII indicate two haplotypes of TaCYP78A5‐2A.