Figure 5.
Role of the TRAPPII complex versus ECHIDNA in hypocotyl/root trade-offs. Seedlings were germinated on ½ MS in the dark (dark) or in the dark with −0.4 MPa water stress (darkW). (A) Col-0 (wild type). (B) BR signaling mutant bin2-3bil1bil2 triple knockout (from Kalbfuß et al., 2022). (C)club-2, a null trappii mutant. (D)trs120-4, a null trappii allele. (E) Null echidna allele, impaired in TGN structure and function. At least three experiments were performed for each line, and a representative one is shown here based on RQ and P values. (F) Representative seedling images of Col-0 (wild type) and mutants shown in A–E. bin2-3bil1bil2 and trs120-4 mutants failed to correctly adjust their hypocotyl and root lengths from dark to darkW conditions. Dotted lines mark the hypocotyl-root junction, whereas arrows point to the end of the root. Scale bar is 1 cm. (G) Normalized response quotient RQratio. Each replicate is represented by a dot. A value of 1 (vertical red line) corresponds to an identical adaptation to darkW conditions as the wild type. Note that the triple bin2-3bil1bil2 knock out, trs120-4 and club-2 had attenuated responses (magenta arrows). (H) Volcano plot with the mean RQratio depicted on the X axis and the median P value of the response on the Y axis (negative log scale; a median of all replicates was used). The area shaded in gray on the red line (green arrow) is where wild-type ecotypes would theoretically map onto the plot. Mutants in the lower left quadrant (peach shading) were considered to have a “confused decision phenotype” (see text). trs120-4 mutants mapped to the lower left quadrant and qualified as decision mutants on two counts: (i) a consistently opposite hypocotyl response (red asterisk in D), (ii) failure to adjust the hypocotyl/root ratio to darkW (the ratio for darkW is the same as for dark in D), translating into a non-significant P value for the ratio response (H). The number (n) of seedlings measured per condition is in gray below the mean ± SD bar graphs. P values were computed with a two-tailed Student’s t test and are represented as follows: *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001; *****: P < 0.00001. Mutant alleles and the corresponding ecotypes are described in Table S2. All data points for A–E are shown in Fig. S7. Related to Figs. S7 and S8.

Role of the TRAPPII complex versus ECHIDNA in hypocotyl/root trade-offs. Seedlings were germinated on ½ MS in the dark (dark) or in the dark with −0.4 MPa water stress (darkW). (A) Col-0 (wild type). (B) BR signaling mutant bin2-3bil1bil2 triple knockout (from Kalbfuß et al., 2022). (C)club-2, a null trappii mutant. (D)trs120-4, a null trappii allele. (E) Null echidna allele, impaired in TGN structure and function. At least three experiments were performed for each line, and a representative one is shown here based on RQ and P values. (F) Representative seedling images of Col-0 (wild type) and mutants shown in A–E. bin2-3bil1bil2 and trs120-4 mutants failed to correctly adjust their hypocotyl and root lengths from dark to darkW conditions. Dotted lines mark the hypocotyl-root junction, whereas arrows point to the end of the root. Scale bar is 1 cm. (G) Normalized response quotient RQratio. Each replicate is represented by a dot. A value of 1 (vertical red line) corresponds to an identical adaptation to darkW conditions as the wild type. Note that the triple bin2-3bil1bil2 knock out, trs120-4 and club-2 had attenuated responses (magenta arrows). (H) Volcano plot with the mean RQratio depicted on the X axis and the median P value of the response on the Y axis (negative log scale; a median of all replicates was used). The area shaded in gray on the red line (green arrow) is where wild-type ecotypes would theoretically map onto the plot. Mutants in the lower left quadrant (peach shading) were considered to have a “confused decision phenotype” (see text). trs120-4 mutants mapped to the lower left quadrant and qualified as decision mutants on two counts: (i) a consistently opposite hypocotyl response (red asterisk in D), (ii) failure to adjust the hypocotyl/root ratio to darkW (the ratio for darkW is the same as for dark in D), translating into a non-significant P value for the ratio response (H). The number (n) of seedlings measured per condition is in gray below the mean ± SD bar graphs. P values were computed with a two-tailed Student’s t test and are represented as follows: *: P < 0.05; **: P < 0.01; ***: P < 0.001; ****: P < 0.0001; *****: P < 0.00001. Mutant alleles and the corresponding ecotypes are described in Table S2. All data points for A–E are shown in Fig. S7. Related to Figs. S7 and S8.

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