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. 2024 Jun 21;121(26):e2321877121. doi: 10.1073/pnas.2321877121

Fig. 1.

Fig. 1.

ChCUC1 is sufficient to increase A. thaliana leaf complexity upon interspecific gene transfer and its expression associates with PIN1 polarity reversals. (A and B) Silhouettes of rosette leaf 8 from wild-type A. thaliana (Columbia-0, Col-0) (A) and C. hirsuta (Oxford; Ox) (B). (C and D) Silhouettes (C) and transgene expression (D) in A. thaliana rosette leaf 8 carrying AtCUC1:V (AtCUC1p::AtCUC1g:Venus). The Inset in (D) shows the vegetative shoot apex. (E and F) Silhouettes (E) and transgene expression (F) in C. hirsuta rosette leaf 8 carrying ChCUC1:V (ChCUC1p::ChCUC1g:Venus). (G and H) Silhouettes (G) and transgene expression (H) in A. thaliana rosette leaf 8 carrying ChCUC1:V (ChCUC1p::ChCUC1g:V). (D, F, and H) Maximum intensity projections of confocal stacks. Green: Venus signal; magenta: cell walls visualized with propidium iodide (PI). (I) Leaf complexity of leaf 8 from the indicated genotypes by measurement of Normalized Difference Margin Complexity [(perimeter contour-perimeter convex hull)/(perimeter contour + perimeter convex hull)]. Letters a and b indicate significant differences by the Kruskal–Wallis test with Dunn’s post hoc test (α = 0.05). ChCUC2:V refers to ChCUC2p::ChCUC2g:V (34). (C–I) Replication: n (phenotypic analysis) ≥ 20 transgenic T1 lines, n (confocal microscopy) = 3 lines. (J–M’) C. hirsuta leaf 5 at different developmental stages showing epidermal expression of ChCUC1:V (ChCUC1p::ChCUC1g:V) and ChPIN1:GFP (ChPIN1p::ChPIN1g:eGFP) projected onto a MorphographX mesh (SI Appendix, Materials and Methods). The white arrows on the leaf margin cells indicate the direction of ChPIN1:GFP polarity. The black arrowheads in (L’) indicate leaf margin cells with ChCUC1:V expression and apical ChPIN1:GFP polarity. The dotted Insets indicate the areas magnified in (J’–M’). D, distal; P, proximal; TL, terminal leaflet; LL, lateral leaflet; iLL, initiating lateral leaflet. n = 3 leaves per stage. The leaf silhouettes were obtained 21 d after sowing. [Scale bars: 1 cm (A–C, E, and G); 100 µm (D, F, and H); 20 µm (J–M’).]