DOI: 10.5061/dryad.f4qrfj745
title: Hibiscus bullseyes reveal mechanisms controlling petal pattern proportions that influence plant-pollinator interactions

description: Colourful flower patterns are key signals to attract pollinators. To
 produce such motifs, plants specify boundaries dividing petals into
 subdomains where cells develop distinctive pigmentations, shapes and
 textures. While some transcription factors and biosynthetic pathways
 behind these characteristics are well-studied, the upstream processes
 restricting their activities to specific petal regions remain enigmatic.
 Here, we unveil that the petal surface of Hibiscus trionum, an emerging
 model featuring a bullseye on its corolla, is pre-patterned as the
 bullseye boundary position is specified long before it becomes visible.
 Using a computational model, we explore how pattern proportions are
 maintained while petals experience a 100-fold size increase. Exploiting
 transgenic lines and natural variants, we show that plants can regulate
 boundary position during the pre-patterning phase or modulate growth
 either side of this boundary later in development to vary bullseye
 proportions. Such modifications are functionally relevant, as buff-tailed
 bumblebees (Bombus terrestris) can reliably identify food sources based on
 bullseye size and prefer certain pattern proportions.
description: See Methods in our manuscript
description: # Hibiscus bullseyes reveal mechanisms controlling petal pattern
 proportions that influence plant-pollinator interactions
 [https://doi.org/10.5061/dryad.f4qrfj745](https://doi.org/10.5061/dryad.f4qrfj745) This contains the data associated with bee behavioural experiments (Figure 7) as well as quantifications associated with characterisation of cell behaviour across the adaxial petal epidermis of H. trionum wild-type and over expression HtTCP4.1 transgenic line as well of H. richardsonii (Figure 1-6). This also contains all the code associated with the computational 1-dimensional model. ## Description of the data and file structure **Data_Source.zip folder contains:** Bumblebees.xlsx file - This file contains the raw data related to our bumblebee behavioural experiments (Fig. 7 and Fig. S8) This folder also contains six files reporting the raw data used to characterise cell behaviour: **boundaries.csv** * tag: describe flower genotype * stage: describe stage of the petal primordia analysed (S0a, S0b, Soc, S1, S2E = Stage 2 early, S2L = Stage 2 Late, S3, S4) * petal_number: number used to identify individual petal * bound: relative position of the boundary along the petal proxino-distal axis (value between 0 and 1, with 0 = base of the petal and 1 = tip of the petal) **Bumblebees.xlsx** * Preferential test tab: * ExpeType: describe the comparison made * Bee: reports the number used to identify each individual * Freq_CUBG: the percentage of the first flower type chosen during the first 10 choices  * Freq_Rich: the percentage of the second flower type chosen during the first 10 choices  * BinaryHtrionum_Rich tab: report the results of the binary test were naive bees were asked to chose between two flower types, equally rewarding * Bee#: reports the number used to identify each individual * Bee color: describe the marking used to identify each individual * Choice per position: report the choice made by the bee at each round, with R = richardsonii and Ht = trionum. * BinaryHtrionumHtTCP4.2 tab: report the results of the binary test were naive bees were asked to chose between two flower types, equally rewarding * Bee#: reports the number used to identify each individual * Bee color: describe the marking used to identify each individual * Choice per position: report the choice made by the bee at each round, with T = HtTCP4.1 and Ht = trionum. * Foraging speed tab: reports the raw data associated with the foraging speed experiment * Bees: describe the color marking used to identify each individual bee * Flowers: describe the flower type used in the experiment * Speed: describe the time (in secondes) taken by the bee to fly between two flowers of the same flower type **CellDivisionEvents.xlsx** * Division_stage0a tab:  * Genotype: genotype of the plant analysed * Stage: describe stage of the petal primordia analysed (S0a) * Petal: describe petal size (in mm) * Ynorm: normalisation along the Y axis * Division_stage0b tab:  * Genotype: genotype of the plant analysed * Stage: describe stage of the petal primordia analysed (S0b) * Petal: describe petal size (in mm) * Ynorm: normalisation along the Y axis **HRichardsonii_PigmentationArea.xlsx** * Genotype: genotype of the plant analysed * Stage: developmental stage at which the petal was analysed * petal: identifier used to identify each individual petal measured * PetalLength: length of the petal (in mm) * Purple_area: area of the purple pigmented region (in mm^2) * Overall_area: reports on the overall area of the entire petal (in mm^2) * Purple_overall_ratio_area: reports on the ratio between purple area and total petal area **HRichardsoniiCellAreaDistributionPDaxis_allStages.xlsx** * name: genotype of the plant analysed * Stage: developmental stage at which the petal was analysed * petal: identifier used to identify each individual petal measured * yNorm: position of the cell analysed along the y-axis * area: cell area (in um^2) * length_majorAxis: length of the cell along the major axis (in um) * length_minorAxis: length of the cell along the minor axis (width, in um) * circularity: circularity value of the cell * aspectRatio: aspect ratio of the cell **HtrionumCellAreaDistributionPDaxis_allStages.xlsx** * name: genotype of the plant analysed * Stage: developmental stage at which the petal was analysed * petal_number: identifier used to identify each individual petal measured * yNorm: position of the cell analysed along the y-axis * area: cell area (in um^2) * length_majorAxis: length of the cell along the major axis (in um) * length_minorAxis: length of the cell along the minor axis (width, in um) * circularity: circularity value of the cell * aspectRatio: aspect ratio of the cell **HtTCP4.1CellAreaDistributionPDaxis_allStages.xlsx** * name: genotype of the plant analysed * Stage: developmental stage at which the petal was analysed * petal_number: identifier used to identify each individual petal measured * yNorm: position of the cell analysed along the y-axis * area: cell area (in um^2) * length_majorAxis: length cell analysed along the major axis of the cell (in um) * length_minorAxis: length of the cell analysed along the minor axis (width, in um) * circularity: circularity value for the cell analysed * aspectRatio: aspect ratio of the cell analysed **1D_TheoreticalModel.zip folder contains:** Code associated with the one-dimensional model. The folder contains three subfolders (data, model_chr and notebook), a README file with instructions, two python files with code (*boundary.py* and *gopetal.py*) and a text file outlining the requirements data must follow to be analysed using those codes. * data subfolder Folder containing the analysed data accompanying this paper. * model_chr subfolder The code for the simulation framework for Chromar, the language used for the model described in the paper, the model itself, and other analysis code related to the results of the simulations. * notebooks subfolder Jupyter notebooks containing the code used to produce the figures in the paper that were produced programmatically. * *boundary.py* Code for calculating the boundary between the two regions. * *go_petal.py* Code for handling the data results as output from MorphoGraphX analysis (data folder). ## Sharing/Access information Code also publicly accessible using the following GitHub link: * [https://gitlab.com/slcu/teamHJ/publications/riglet_and_zardilis_etal_2024/](https://gitlab.com/slcu/teamHJ/publications/riglet_and_zardilis_etal_2024/ "https://gitlab.com/slcu/teamhj/publications/riglet_and_zardilis_etal_2024/")