Fig. 4

Functional validation of candidate gene. A The morphology of knockout (KO) lines and Kitaake (KTK) at the mature stage. B The deep-green leaf phenotype of KO lines at maturity. C Comparison of leaves chlorophyll a and chlorophyll b contents of WT and KO1 at the maturity stage. Mean and SD were obtained from three independent experiments. Comparison of plant height (D), number of tillers (E), kernels per spike (F), 1000-grains weight (G) and seed setting rate (H) between KTK and KO1 respectively. I The morphology of complementary lines (CP) at maturity. J The leaf blade color of WT, hcc1 and CP1 at maturity. K Comparison of leaves chlorophyll a and chlorophyll b contents at maturity stage between WT, hcc1 and CP1. Mean and SD were obtained from three independent experiments. L–P showed plant height, number of tillers, kernels per spike, 1000-grains weight and seed setting rate between WT and CP1 respectively. Mean and SD were obtained from three independent measurements in C and K. Data presented in D–H, L–P are the average of n = 10 plants. Statistical analysis was performed using two-tailed Student’s t-test, *, **, ***, ****indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, “ns” indicates no significant difference. Scale bar in A, I = 20 cm, and in B, J = 2 cm