Exploring conservation of cellular-level traits in shade avoidance syndrome among species 5 179 ► Figure 5.7. Soybean (Glycine max) stem elongation in response to FR. G. max seedlings were subjected to one week of WL+FR treatment versus WL treatment. (a)Phenotype of soybean in FR or WL. The phenotyping data include measurements of (b) stem length, (c) hypocotyl length, (d) internode1 length, (e) hypocotyl diameter, and (f) internode1 diameter. The data are presented as mean ± SEM, and different letters indicate significant differences between treatments based on ANOVA analysis with Tukey’s post hoc test (P<0.05). There are 12 biological replicates, and the experiment was repeated twice. The following microscopy analysis results aspects are examined:(g) cell layers thickness from cross sections, (h) number of cell layers from cross section, (i) cell length from longitudinal sections, and (j) vasculature bundle area from cross section. Asterisks in the figure indicate the significance of differences between WL and WL+FR treatments, with the following significance levels: * p≤0.05; ** p≤0.01; *** p≤0.001. Error bars represent the standard error (SE), and the sample size for each measurement is n=40. (k) Negative logarithm of p values for cell type feature comparison of WL vs WL+FR measurement in soybean. 5.2.4 Minimal FR responsiveness in basil and pea We also tested the FR-responsiveness of species that did not have such clear FR responses. Firstly, O. basilicum (basil) from Lamiaceae (mint family) exhibited a mild internode elongation response to FR light, though this response did not manifest at the cellular level (Figure 5.8). When treated with WL+FR, basil plants displayed differences only after two weeks of FR treatment; no noticeable elongation occurred after one week. Notably, there were also no disparities in the diameter of the first internode (Figure 5.8). Regarding basil’s cellular characteristics, pith cell elongation in response to FR light was not observed either. Secondly, another legume, pea (Pisum sativum) showed no significant differences in stem length or diameter between the two treatments (Figure 5.9). Diameter measurements were taken from at least three locations along the stem’s length and the average was calculated. Pea stem cross-sections revealed a tetrarch symmetry leading to an X-shaped pith, necessitating specific measurement directions (Figure 5.9). Pith measurements focused on the pith area, predominantly composed of cells stained with dense red color, inside of the X-shaped area between vascular bundles (Figure 5.9h). We found no differences between treatments for the measured cell types (Figure 5.9 i,k,l,m).
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