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Figure 8. PAP also acts as an ABA secondary messenger in Arabidopsis seeds.(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.DOI:
http://dx.doi.org/10.7554/eLife.23361.016Figure 8—figure supplement 1. Role of SAL1 and PAP in regulating seed germination.Wild type seed differentially regulates SAL1 transcript and protein abundance to accumulate more PAP than leaves. (A) SAL1 expression is the lowest during seed development and in dry seed (red boxes), and highest when the seed is imbibed (black arrow). Data visualized from publically-deposited microarray data using eFP Browser. (B) Analysis of publically-deposited microarray data in Genevestigator using the Genevisible tool reveals that the top four environmental changes that exert the highest degree of change in SAL1 transcript abundance, out of >3000 perturbations, are the transition from dry seed to germinating seed. This suggests that the transcriptional repression of SAL1 and the associated PAP accumulation in seed are key aspects of seed development and dormancy.DOI:
http://dx.doi.org/10.7554/eLife.23361.017 Image published in: Pornsiriwong W et al. (2017) © 2017, Pornsiriwong et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |