High ambient temperature affects flowering time, floret, and seed number per main spike in barley. Days to flower (A), the number of florets (B), and the number of seeds per main spike (C) under control (blue; 20°C/16°C, day/night) and high ambient temperatures (pink; 28°C/24°C, day/night) in the spring barley varieties Bowman and Scarlett and the derived ILs Bowman(eam8) (Hvelf3), Bowman(Ppd-H1), S42-IL107 (Ppd-H1), and S42-IL176 (Hvvrn1). Flowering time, floret, and seed number were recorded for 20 plants per genotype and treatment under LDs (16 h light/8 h night). N.F indicates nonflowering plants. Statistical differences were calculated by a two-factorial ANOVA and a post-hoc Tukey’s multiple comparison test: *P < 0.05, **P < 0.01, ***P < 0.001; n.s = nonsignificant.