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. 2024 Apr 4;15(1):2912.
doi: 10.1038/s41467-024-47325-1.

Two orthogonal differentiation gradients locally coordinate fruit morphogenesis

Andrea Gómez-Felipe  1 , Elvis Branchini  1 , Binghan Wang  1 , Marco Marconi  2   3 , Hana Bertrand-Rakusová  1 , Teodora Stan  1 , Jérôme Burkiewicz  1 , Stefan de Folter  4 , Anne-Lise Routier-Kierzkowska  1 , Krzysztof Wabnik  2   3 , Daniel Kierzkowski  5
Affiliations

Two orthogonal differentiation gradients locally coordinate fruit morphogenesis

Andrea Gómez-Felipe et al. Nat Commun. .

Abstract

Morphogenesis requires the coordination of cellular behaviors along developmental axes. In plants, gradients of growth and differentiation are typically established along a single longitudinal primordium axis to control global organ shape. Yet, it remains unclear how these gradients are locally adjusted to regulate the formation of complex organs that consist of diverse tissue types. Here we combine quantitative live imaging at cellular resolution with genetics, and chemical treatments to understand the formation of Arabidopsis thaliana female reproductive organ (gynoecium). We show that, contrary to other aerial organs, gynoecium shape is determined by two orthogonal, time-shifted differentiation gradients. An early mediolateral gradient controls valve morphogenesis while a late, longitudinal gradient regulates style differentiation. Local, tissue-dependent action of these gradients serves to fine-tune the common developmental program governing organ morphogenesis to ensure the specialized function of the gynoecium.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1. Cellular growth patterns underlying gynoecium development.
a, b, Heat-maps of area expansion (a) and growth anisotropy (b) for the Arabidopsis thaliana gynoecium. c Lineage tracing of style (green), valves (blue), and replum (yellow) between 5 and 13 DAI. d, e Quantifications of area expansion (d) and growth anisotropy (e) in different regions of the developing gynoecium (n = 136 cells at 3 DAI; n = 346 cells at 4 DAI; n = 94 (style), 516 (valves), 201 (replum) cells at 5 DAI; n = 109 (style), 836 (valves), 347 (replum) cells at 6 DAI; n = 190 (style), 1421 (valves), 568 (replum) cells at 7 DAI; n = 298 (style), 2459 (valves), 913 (replum) cells at 8 DAI; n = 432 (style), 3921 (valves), 1425 (replum) cells at 9 DAI; n = 584 (style), 7039 (valves), 2078 (replum) cells at 10 DAI; n = 727 (style), 10,933 (valves), 2789 (replum) cells at 11 DAI; n = 767 (style), 14,579 (valves), 3545 (replum) cells at 12 DAI; n = 532 (style), 10,779 (valves), 2778 (replum) cells at 13 DAI; three independent time-lapse series). The boxplots represent a range between the first and the third quartile and the whiskers include 95% of the values. Lines represent the median and dots represent the mean. DAI days after gynoecium initiation. Scale bars, 100 µm. See also Supplementary Fig. 1 and Supplementary Movies 1 and 2.
Fig. 2
Fig. 2. Two orthogonal gradients occur during gynoecium development.
a, b Heat-maps of the averaged cellular growth along longitudinal (a) and mediolateral (b) axes of the gynoecium. c Quantification of the cellular growth along the longitudinal (left) and mediolateral (right) axis of the valve as a function of the distance from the valve base or from the replum (n = 137 cells at 3–4 DAI; n = 283 cells at 4–5 DAI; n = 408 cells at 5–6 DAI; n = 789 cells at 6–7 DAI; n = 1365 at 7–8 DAI). d Heat-maps of averaged cellular growth along the longitudinal axis of the style. e Quantification of the cellular growth along the longitudinal axis of the style as a function of the distance from the gynoecium tip (n = 207 cells at 8–9 DAI; n = 207 cells at 9–10 DAI; n = 258 cells at 10–11 DAI; n = 297 cells at 11–12 DAI; n = 298 cells at 12–13 DAI). f Stomata distribution in the valves. Replum in orange, valve in white, stomata in brown, base of the valve in green. g Quantification of stomatal distribution as a function of the distance from the valve base (left) or from the replum (right) (n = 6 stomata at 10 DAI; n = 58 stomata at 11 DAI; n = 517 stomata at 12 DAI; n = 605 stomata at 13 DAI; n = 1209 stomata at 14 DAI; three independent time-lapse series). DAI days after gynoecium initiation. For scatter plots, the distance was normalized. Dots in c, e indicate each growth value, lines represent the average and shaded areas represent standard deviation (SD). Scale bars, 100 µm in (a, b) and 50 µm in (d). See also Supplementary Figs. 1 and 2 and Supplementary Movie 3.
Fig. 3
Fig. 3. Auxin patterning during gynoecium development.
a–k Expression patterns of pDR5v2::nls-3xVenus (a, b), pYUC4::3xNLS-GFP (c, d), pPIN1::PIN1-GFP (e–g), pPIN3::PIN3-GFP (h, i), and pPIN7::PIN7-GFP (j, k) in the A. thaliana gynoecium. Virtual cross sections shown in (b, d, f) are in the middle of the gynoecium. g, i, k Heat-maps represent the intensity of the PIN1-GFP (g), PIN3-GFP (i), and PIN7-GFP (k) signal in the L1 epidermal layer (one side of the gynoecium tops are shown for PIN1 and the style is shown for PIN3 and PIN7). Insets indicate membrane localization and arrows indicate plausible orientation of PIN polarization. Scale bars, 100 µm in (a, c, e, h, j) and 50 µm in (b, d, f, g, i, k). See also Supplementary Fig. 3.
Fig. 4
Fig. 4. Removing valves abolishes the mediolateral and expands the longitudinal gradients.
a, b Heat maps of averaged area expansion (a) and cell sizes (b) in A. thaliana gynoecium one week after treatment with the NPA. c, d Quantification of cellular growth (c) and cell sizes (d) as a function of the distance from the gynoecium base after NPA treatment (n = 889 cells at 5 DAI; n = 1234 cells at 6 DAI; n = 1460 cells at 7 DAI; n = 1463 cells at 8 DAI; n = 1476 cells at 9 DAI; n = 1478 cells at 10 DAI; three independent time-lapse series). e, f Stomata distribution (e) and quantification of stomatal distribution along the distance from the gynoecium base (f) after NPA treatment (n = 3 stomata at 8 DAI; n = 32 stomata at 9 DAI; n = 125 stomata at 10 DAI; three independent time-lapse series). g–k Expression patterns of PIN1-GFP (g–i) and PIN3-GFP (j–k) in gynoecia after NPA treatment. Heat maps represent the intensity of the PIN1-GFP (i) and PIN3-GFP (k) signals in the L1 layer. Insets indicate membrane localization; arrows indicate plausible orientation of PIN polarization. For plots, the distance was normalized, lines represent the average and shaded areas represent standard deviation (SD). DAI days after organ initiation. Scale bars, 100 µm (a, b, g, h, j) and 50 µm (i, k). See also Supplementary Figs. 5 and 6, Supplementary Movie 4.
Fig. 5
Fig. 5. Introducing carpel identity into sepal reorients organ differentiation gradients.
a, b, Wild-type (a) and ap2-7 mutant (b) sepals. c, f Heat maps of averaged area expansion (c, e) and cell sizes (d, f) of wild-type (c, d) and ap2–7 mutant (e, f) sepals. g, h Quantifications of area expansion (g) and cell sizes (h) of wild-type sepal (n = 354 cells at 3 DAI; n = 707 cells at 4 DAI; n = 832 cells at 5 DAI; n = 1803 cells at 6 DAI; n = 2457 cells at 7 DAI; n = 3145 cells at 8 DAI; three independent time-lapse series) and ap2–7 mutant sepal (n = 73 cell at 2 DAI; n = 132 cells at 3 DAI; n = 375 cells at 4 DAI; n = 566 cells at 5 DAI; n = 881 cells at 6 DAI; n = 1299 cells at 7 DAI; n = 2135 cells at 8 DAI; n = 3279 cells at 9 DAI; n = 1294 cells at 10 DAI; three independent time-lapse series) along the longitudinal axis of the organ. For plots, the distance was normalized, lines represent the average and shaded areas represent standard deviation (SD). DAI days after organ initiation. Scale bars, 500 µm (a, b), 100 µm (c–f). See also Supplementary Fig. 7 and Supplementary Movie 5.
Fig. 6
Fig. 6. Removing style identity and reducing CMM expands the longitudinal gradient.
a Gynoecium of the crc-1 spt-12 double mutant at 10 DAI (top); longitudinal cut (bottom) shows ovule-like structures. b Heat maps of averaged area expansion of crc-1 spt-12. c Stomata distribution of crc-1 spt-12. d, e Quantification of stomatal distribution as a function of the distance from the valve base (e) or from the medial region (d) of crc-1 spt-12 (n = 6 stomata at 5 DAI; n = 15 stomata at 6 DAI; n = 61 stomata at 7 DAI; n = 309 stomata at 8 DAI; n = 815 stomata at 9 DAI; three independent time-lapse series). f Schematic representation of the influence of the tissue identity and marginal meristematic activity on developmental gradients. Removing valve’s identity (after NPA treatment) abolishes the mediolateral and expands the longitudinal gradients. Removing style identity and reducing CMM (in crc-1 spt-12 mutant), accelerates and expands the longitudinal differentiation gradient. Introducing valve identity and CMM into sepal reorients organ differentiation gradients. Orange indicates CMM; green, cell differentiation status; arrow, the orientation of cell differentiation progression. DAI days after organ initiation. Scale bars, 500 µm (a, b), 100 µm (c–f). See also Supplementary Movie 6.
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References

    1. Rogers KW, Schier AF. Morphogen gradients: from generation to interpretation. Annu. Rev. Cell Dev. Biol. 2011;27:377–407. doi: 10.1146/annurev-cellbio-092910-154148. - DOI - PubMed
    1. Briscoe J, Small S. Morphogen rules: design principles of gradient-mediated embryo patterning. Development. 2015;142:3996–4009. doi: 10.1242/dev.129452. - DOI - PMC - PubMed
    1. Green JBA, Sharpe J. Positional information and reaction-diffusion: two big ideas in developmental biology combine. Development. 2015;142:1203–1211. doi: 10.1242/dev.114991. - DOI - PubMed
    1. Echevin E, et al. Growth and biomechanics of shoot organs. J. Exp. Bot. 2019;70:3573–3585. doi: 10.1093/jxb/erz205. - DOI - PubMed
    1. Moon J, Hake S. How a leaf gets its shape. Curr. Opin. Plant Biol. 2011;14:24–30. doi: 10.1016/j.pbi.2010.08.012. - DOI - PubMed

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