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. 2020 Aug 6;32(10):3346–3369. doi: 10.1105/tpc.20.00027

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© 2020 American Society of Plant Biologists. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 10. — Suppression by Ca²⁺ of the Severe Etiolated Hypocotyl Phenotype of glcat14a/b/e.

(A) Nine-day-old dark-grown seedlings from wild-type and segregating glcat14a⁺⁻/b/e plants grown on basal MS medium and MS medium supplemented with 2, 6, 12, or 24 mM CaCl₂. Arrowheads indicate glcat14a/b/e triple mutants. Bars = 0.5 cm.

(B) Box-plot representation of the lengths of hypocotyls from (A). Gray dots represent the value of single measurements, and black dots are outliers. The cross represents the mean value of three biological replicates, and the horizontal line represents the median. Hypocotyls from glcat14a⁺⁻/b/e and glcat14a/b/e were not fully discernible when grown on MS medium supplemented with 12 and 24 mM CaCl₂ and were therefore not used for statistical analysis. n = 50 wild-type, 180 glcat14a⁺⁻/b/e, and 50 glcat14a/b/e hypocotyls per treatment per replicate.

(C) Hypocotyl lengths of 9-d-old dark-grown wild-type, glcat14a/b, and glcat14a/b/d seedlings grown on basal MS medium and MS medium supplemented with 12 mM CaCl₂. n = 120 per line per replicate.

(D) Nine-day-old dark-grown wild-type, qua1, and qua2 seedlings grown on basal MS medium (1) and MS medium supplemented with 12 mM (2) or 24 mM (3) CaCl₂. Bar = 0.5 cm.

(E) Quantification of the length of hypocotyls from (D). Values are means ± sd of three biological replicates. n = 25 per line per replicate.

(F) Nine-day-old dark-grown wild-type, cop1, and det1 seedlings grown on basal MS medium and MS medium supplemented with 12 and 24 mM CaCl₂. Bars = 0.25 cm.

For (B), asterisks indicate significant differences between glcat14a⁺⁻/b/e and glcat14a/b/e as defined by two-way ANOVA followed by Sidak’s multiple comparison test (asterisks above the boxes). Significant differences between MS medium and increased CaCl₂ were defined by one-way ANOVA followed by Tukey’s multiple comparison test. For (C) and (E), significant differences between lines (C) and treatments (E) were defined by one-way ANOVA followed by Tukey’s multiple comparison test or Student’s t test for two-sample comparisons: **, P < 0.01 and ****, P < 0.0001. NS, not significant.