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Section: New Results

Multi-scale models and analysis: from cells to plant architecture (and back)

Transport model in roots

Participants : Mikaël Lucas [IRD] , Christophe Pradal, Christophe Godin, Christophe Maurel [BPMP] .

This research theme is supported by the ANR project HydroRoot.

A model of Arabidopsis thaliana root hydraulics at the cellular level was developped in the OpenAlea modeling platform. The model relies on the integration throughout root architecture of elementary hydraulic components. Each component integrates local radial and axial water flows. Axial hydraulic conductivity is calculated according to Poiseuille’s law, based on local size of xylem vessels. Radial hydraulic conductivity is determined in part by aquaporin activity and was set constant throughout root architecture in the first model versions. In its current state, the model is parameterized using architectural, tissular and physiological data that were experimentally determined in the Aquaporin group at BPMP. The architectural reconstruction of the root system is based on a tridimensional multi-scale tree graph (MTG). The current model is capable of predicting the water flow that is transported by a root system in the standard experimental conditions used in the Aquaporin group. This model was used to perform sensitivity analyses and determine the respective contributions to root hydraulic dynamics of various biological parameters (axial and radial hydraulic conductivites, root architecture). One major finding is that the root hydraulic conductivity (Lpr) computed from the model is highly dependent on root architecture. This is due to the limiting role of axial (xylem) conductance, one feature that had been neglected in previous representations of root water transport. The radial hydraulic conductivity may primarily be limiting in conditions of Lpr inhibition, since its increase from values in control roots has marginal effects on Lpr. A new set of experimental data including root diameter repartitions in wild-type plants, and xylem vessel diameters in mutants with altered xylem morphology (irx3, esk1) will be used to implement the model. Root cell hydraulic conductivities will also be measured in these and aquaporin mutant phenotypes. Our aim is to check whether, based on anatomical and morphological data, the model can properly predict the radial hydraulic conductivity of these genotypes.

Transport in fruits

Participants : Mik Cieslak, Nadia Bertin [Inra, Avignon] , Frédéric Boudon, Christophe Godin, Michel Genard [Inra, Avignon] , Christophe Goz-Bac [Université Montpellier 2] .

This research theme is supported by the Agropolis project Fruit3D.

Understanding the controlling factors of fruit quality development is challenging, because fruit quality results from the interplay between physical and physiological processes that are under the control of genes and the environment. Although process-based models have been used to make significant progress in understanding these factors, they ignored to a large extent the shape and internal structure of the fruit.

To help characterizing effects of fruit shape and internal structure on quality, the creation of a 3D virtual fruit model that integrates fruit structure and function with growth governed by environmental inputs has been investigated. For this, a modeling pipeline has been developed that includes the following steps: creation of a 3D volumetric mesh of the internal fruit structure, including vasculature (see section 3 ). Based on previous compartment models of fruit physiology developed at Avignon, we have then developed models of water and carbon transport that have been coupled with the 3D model of fruit. In the 3D model, different equations are describing the transport between adjacent regions of the fruit represented as a 3D mesh. The integration through space and time is carried out using a standard integration scheme (Runge-Kutta of order 4).

This approach has been applied to study tomato fruit (Solanum lycopersicum) by constructing 3D volumetric meshes from different sources (images of perpendicular fruit slices and MRI data), and integrating water and carbon transport processes into these meshes. To illustrate the tomato model, a simulation of one season of the fruit’s growth has been performed and its results compared with an already published process-based tomato fruit model. We first showed that our spatialized model is compliant with classical results of the abstract process-based models but also provides additional information on the internal heterogeneity of the fruit, such as a gradient in sugar concentration. Once the model is calibrated and evaluated, our approach will be suitable for studying the effects of internal fruit heterogeneity and overall shape on fruit quality development.

Figure 4. Virtual models of peaches reconstructed from images with simulated vasculatur to simulate carbon and water transport in the fruit
IMG/virtualfruit.png

Analyzing root growth and branching

Participants : Beatriz Moreno Ortega, Sixtine Passot, Yann Guédon, Laurent Laplaze [IRD, DIADE] , Mikaël Lucas [IRD, DIADE] , Bertrand Muller [INRA, LEPSE] .

This research theme is supported by two PhD programmes.

New 2D and 3D root phenotyping plateforms are emerging with associated image analysis toolbox (e.g. SmartRoot). The analysis of complex root phenotyping data is thus a new challenge in developmental biology.

We aim at developing a pipeline of methods for analyzing root systems at three scales:

  1. tissular scale to identify and characterize the meristem, elongation and mature zones along a root using piecewise heteroscedastic linear models.

  2. individual root scale to analyze the dynamics of root elongation

  3. root system scale to analyze the branching structure.

This pipeline of analysis methods will be applied to different species (maize, millet and arabidopsis) and for different biological objectives (study of genetic diversity for millet and of metabolic and hormonal controls of morphogenesis for maize).

Analyzing shoot and leaf elongation

Participants : Maryline Lièvre, Yann Guédon, Christine Granier [INRA, LEPSE] .

This research theme is supported by one PhD programme.

The analysis of phenotyping data coming from automated platforms such as PHENOPSIS often focuses on the growth of a leaf at a given rank along the stem. We aim at developing a pipeline of methods for analyzing the growth of arabidopsis shoot at three scales:

  1. tissular scale using a probabilistic model of endoreduplication for modeling the distribution of the leaf epidermis cell surfaces. Endoreduplication, which is a replication of the nuclear genome in the absence of cell division that leads to elevated nuclear gene content, strongly affects the leaf epidermis cells of arabidopsis.

  2. organ scale using nonlinear regression model for analyzing the growth of each successive leaf.

  3. shoot scale: The outputs of the analyses at the tissular and organ scales will be summarized as multivariate sequences along the shoots characterizing each successive leaf. These sequences will be augmented by supplementary morphological variables characterizing leaf shape and properties (e.g. presence/absence of trichomes). These sequences will be globally analyzed in order to take into account plant ontogeny and in particular the successive developmental stages before the floral transition for the wild type and selected mutants of arabidopsis.

Analyzing perturbations in Arabidopsis thaliana phyllotaxis

Participants : Christophe Godin, Yann Guédon, Yassin Refahi, Etienne Farcot, Teva Vernoux, Fabrice Besnard [RDP, ENS] .

This research theme is supported by iSAM.

The geometric arrangement of lateral organs along plant stems, named phyllotaxis, shows a variety of striking patterns with remarkable regularities and symmetries. This has interested biologists, physicists, mathematicians and computer scientists for decades. These studies have lead to a commonly accepted standard interpretation of phyllotaxis that postulates that organs inhibit the formation of new organs in their vicinity. At a molecular scale, these inhibitory fields have been shown to result from the spatio-temporal distribution of the plant hormone auxin. This model theoretically explains a large part of the diversity of phyllotactic patterns observed in plants.

The cytokinin hormones are known to play a significant role in the regulation of phyllotaxis. Fabrice Besnard and Teva Vernoux realized that Arabidopsis thaliana ahp6 mutants, which are perturbed in the cytokinin signaling pathway, showed unusual chaotic perturbations of the phyllotaxis at macroscopic level.

In order to characterize these perturbations, we designed a pipeline of models and methods which relies of combinatorial and statistical techniques. Using this pipeline of methods, we have shown that the perturbation patterns in both wild-type and mutant plants can be explained by permutations in the order of insertion along the stem of 2 or 3 consecutive organs. The number of successive synchronized organs between two permutations reveals unexpected patterns that depend on the nature of the preceding permutation (2- or 3-permutation). We identified significant individual deviations of the level of baseline segments with reference to 137.5°, which confirms theoretical model predictions. Finally, we highlighted a marked relationship between permutation of organs and defects in the elongation of the internodes in between these organs.

We then looked at the origin of these permutations using confocal microscopy and realized that organs were in fact frequently co-initiated in the mutant, leading after development randomly in half of the cases to permutations. We concluded that the mutant is actually perturbed in the time between consecutive organ initiation (i.e. the plastochrone), while relative angular positions are not affected. After closer inspection, we realized that the mutated gene encode a protein diffusing from the organs and creating a field around the organs that regulates the plastochrone. We could demonstrate that in the mutant, the absence of this field lead to co-initiations and subsequently to the observed permutations.

Altogether, this study sheds a new light on our interpretation of phyllotaxis, revisiting the standard model and suggesting that several fields based on auxin and cytokinin with different properties are required to provide robustness to phyllotaxis. An overview of this work has been published in the journal Nature in December online [13] . Methodological developments were published more extensively in [20] )