Extended Data Fig. 10: Mutation of active sites of ECS1 and ECS2 endopeptidases leads to polytubey.
From: Fertilized egg cells secrete endopeptidases to avoid polytubey
a, Mutation of active sites of ECS1 and ECS2 (Extended Data Fig. 2a) led to reduced proteolytic activity. Proteolytic activities of recombinant WT and mutant version of ECS towards cleavage of fluorogenic peptide 3 (Extended Data Fig. 6) were measured respectively. Data represent mean ± s.d. of three independent experiments. b, Mutant version of ECS could not recover the polytubey phenotype of ecs1 ecs2 double mutant. Proportions of polytubey in ecs1 ecs2 double mutants and different transgenic lines were determined at 24 HAP (n = 308 for WT; 302 for ecs1 ecs2; 309 for ECS1; 319 for ECS1D103N D324N-1; 302 for ECS1D103N D324N-2; 309 for ECS2; 321 for ECS2D103N D326N-1; 300 for ECS2D103N D326N-2). Data represent the mean ± s.d. Same letters indicate lack of significant differences according to the Tukey–Kramer multiple comparison test (one-way ANOVA between groups; P = 7.82 × 10−18, F = 50.24). P < 0.05 was considered as significant. c, Representative images showing multiple pollen tubes entrance in different transgenic lines as indicated. Arrows indicate pollen tubes (pt). Scale bars, 100 μm.
