(A) PAP levels in wild type and sal1-6 leaves and seeds. Data are means ± SEM, n = 3. Significantly different groups (p<0.05) are shown as a, b, c. Western blot of total protein probed with anti-SAL1, together with rSAL1 recombinant protein control. (B) Germination rates (radicle emergence) of wild type and sal1-6 seeds harvested fresh and plated straight onto 0.8% agarose without prior storage, stratification or sterilization. Values show averages of five to eight plates containing at least 70 seeds per plate ± SEM. Significant differences at p<0.001 (***) or at p<0.05 (*) compared to wild type are shown. (C) Germination rates (radicle emergence) of wild type and sal1-6 non-stratified seeds that were after-ripened at room temperature for one month post-harvest. Values show averages of five plates containing at least 70 seeds per plate ± SEM. Significant differences at p<0.001 (***), p<0.01 (**) or p<0.05 (*) compared to wild type are shown. (D) Germination rates of mature dry wild type and sal1-6 seeds sterilised and sown on MS plates with appropriate chemicals added before stratification and growth. At least 70 seeds were sown in a plate per genotype and eight replicates per genotype per treatment were prepared. Data are means ± SD. Significant difference groups (p<0.05) are shown (a, b, c). (E) Germination rates of wild type, abi1-1 and abi1-1 sal1-8 in the presence of 1 µM ABA or ABA + 0.5 µM of a GA biosynthesis inhibitor: paclobutrazol (PAC). Bar graphs represent the average of two independent experiments (n = 90 seeds per genotype per experiment) ± SEM. Significant difference (p<0.05, *) compared to respective controls. n.s: not significant. (F) Germination rates of WT and abi1-1 seeds under combinatorial treatments of ABA (1, 2 µM), paclobutrazol (0.5 µM) and PAP (10, 50, 100, 500, 1000 µM), shown as a heat map. Multivariate analysis of the complete dataset shows highly significant treatment interactions (table) and were used to calculate significant difference groups (a, b, c, d, e; p<0.05) within a genotype. (G) Subset result of (F) showing individual germination rates for WT and abi1-1 in response to specific combinations of ABA, paclobutrazol and PAP. For both (F) and (G) n = 70 seeds x 5 plates = 350 seeds per genotype per combinatorial treatment.