Exposure to high nitrogen triggered a genotype-dependent modulation of cell and root hydraulics, which can involve aquaporin regulation.

Root nitrogen acquisition has been proposed to be regulated by mass flow, a process by which water flow brings nutrients to the root surface, depending on a concerted regulation of the root hydraulic properties and stomatal conductance. As aquaporins play an important role in regulating transcellular water flow, we aimed at evaluating the short-term effect of high nitrogen (HN) availability on the dynamics of hydraulic parameters at both the root and cell level and the regulation of aquaporins. The effect of short-term HN (8 mM NO3 - ) treatment was investigated on 12 diverse 15-day-old maize genotypes. Root exposure to HN triggered a rapid (<4 h) increase in the root hydraulic conductivity (Lpr ) in seven genotypes while no Lpr variation was recorded for the others, allowing the separation of the genotypes into two groups (HN-responsive and HN-nonresponsive). A remarkable correlation between Lpr and the cortex cell hydraulic conductivity (Lpc ) was observed. However, while differences in gas exchange parameters were also observed, the variations were genotype-specific and not always correlated with the root hydraulic parameters. We then investigated whether HN-induced Lpr variations were linked to the activity and regulation of plasma membrane PIP aquaporins. While some changes in PIP mRNA levels were detected, this was not correlated with the protein levels. On the other hand, the rapid variation in Lpr observed in the B73 genotype was correlated with the PIP protein abundance in the plasma membrane, highlighting PIP posttranslational mechanisms in the short-term regulation of root hydraulic parameters in response to HN treatment.

GRANAR, a Computational Tool to Better Understand the Functional Importance of Monocotyledon Root Anatomy.

Root hydraulic conductivity is a limiting factor along the water pathways between the soil and the leaf, and root radial conductivity is itself defined by cell-scale hydraulic properties and anatomical features. However, quantifying the influence of anatomical features on the radial conductivity remains challenging due to complex time-consuming experimental procedures. We present an open-source computational tool, the Generator of Root Anatomy in R (GRANAR; http://granar.github.io), that can be used to rapidly generate digital versions of contrasted monocotyledon root anatomical networks. GRANAR uses a limited set of root anatomical parameters, easily acquired with existing image analysis tools. The generated anatomical network can then be used in combination with hydraulic models to estimate the corresponding hydraulic properties. We used GRANAR to reanalyze large maize (Zea mays) anatomical datasets from the literature. Our model was successful at creating virtual anatomies for each experimental observation. We also used GRANAR to generate anatomies not observed experimentally over wider ranges of anatomical parameters. The generated anatomies were then used to estimate the corresponding radial conductivities with the hydraulic model MECHA (model of explicit cross-section hydraulic architecture). Our simulations highlight the large importance of the width of the stele and the cortex. GRANAR is a computational tool that generates root anatomical networks from experimental data. It enables the quantification of the effect of individual anatomical features on the root radial conductivity.

Spatially Resolved Root Water Uptake Determination Using a Precise Soil Water Sensor.

To answer long-standing questions about how plants use and regulate water, an affordable, noninvasive way to determine local root water uptake (RWU) is required. Here, we present a sensor, the soil water profiler (SWaP), which can determine local soil water content (θ) with a precision of 6.10-5 cm3 ⋅ cm-3, an accuracy of 0.002 cm3 ⋅ cm-3, a temporal resolution of 24 min, and a one-dimensional spatial resolution of 1 cm. The sensor comprises two copper sheets, integrated into a sleeve and connected to a coil, which form a resonant circuit. A vector network analyzer, inductively coupled to the resonant circuit, measures the resonance frequency, against which θ was calibrated. The sensors were integrated into a positioning system, which measures θ along the depth of cylindrical tubes. When combined with modulating light (4-h period) and resultant modulating plant transpiration, the SWaP enables quantification of the component of RWU distribution that varies proportionally with total plant water uptake, and distinguishes it from soil water redistribution via soil pores and roots. Additionally, as a young, growing maize (Zea mays) plant progressively tapped its soil environment dry, we observed clear changes in plant-driven RWU and soil water redistribution profiles. Our SWaP setup can measure the RWU and redistribution of sandy-soil water content with unprecedented precision. The SWaP is therefore a promising device offering new insights into soil-plant hydrology, with applications for functional root phenotyping in nonsaline, temperature-controlled conditions, at low cost.

Modification of the Expression of the Aquaporin ZmPIP2;5 Affects Water Relations and Plant Growth.

The plasma membrane intrinsic protein PIP2;5 is the most highly expressed aquaporin in maize (Zea mays) roots. Here, we investigated how deregulation of PIP2;5 expression affects water relations and growth using maize overexpression (OE; B104 inbred) or knockout (KO; W22 inbred) lines. The hydraulic conductivity of the cortex cells of roots grown hydroponically was higher in PIP2;5 OE and lower in pip2;5 KO lines compared with the corresponding wild-type plants. While whole-root conductivity decreased in the KO lines compared to the wild type, no difference was observed in OE plants. This paradox was interpreted using the MECHA hydraulic model, which computes the radial flow of water within root sections. The model hints that the plasma membrane permeability of the cells is not radially uniform but that PIP2;5 may be saturated in cell layers with apoplastic barriers, i.e. the endodermis and exodermis, suggesting the presence of posttranslational mechanisms controlling the abundance of PIP in the plasma membrane in these cells. At the leaf level, where the PIP2;5 gene is weakly expressed in wild-type plants, the hydraulic conductance was higher in the PIP2;5 OE lines compared with the wild-type plants, whereas no difference was observed in the pip2;5 KO lines. The temporal trend of leaf elongation rate, used as a proxy for that of xylem water potential, was faster in PIP2;5 OE plants upon mild stress, but not in well-watered conditions, demonstrating that PIP2;5 may play a beneficial role in plant growth under specific conditions.

Going with the Flow: Multiscale Insights into the Composite Nature of Water Transport in Roots.

As water often limits crop production, a more complete understanding of plant water capture and transport is necessary. Here, we developed MECHA, a mathematical model that computes the flow of water across the root at the scale of walls, membranes, and plasmodesmata of individual cells, and used it to test hypotheses related to root water transport in maize (Zea mays). The model uses detailed root anatomical descriptions and a minimal set of experimental cell properties, including the conductivity of plasma membranes, cell walls, and plasmodesmata, which yield quantitative and scale-consistent estimations of water pathways and root radial hydraulic conductivity (k r). MECHA revealed that the mainstream hydraulic theories derived independently at the cell and root segment scales are compatible only if osmotic potentials within the apoplastic domains are uniform. The results suggested that the convection-diffusion of apoplastic solutes explained most of the offset between estimated k r in pressure clamp and osmotic experiments, while the contribution of water-filled intercellular spaces was limited. Furthermore, sensitivity analyses quantified the relative impact of cortex and endodermis cell conductivity of plasma membranes on root k r and suggested that only the latter contributed substantially to k r due to the composite nature of water flow across roots. The explicit root hydraulic anatomy framework brings insights into contradictory interpretations of experiments from the literature and suggests experiments to efficiently address questions pertaining to root water relations. Its scale consistency opens avenues for cross-scale communication in the world of root hydraulics.

Connecting the dots between computational tools to analyse soil-root water relations.

In recent years, many computational tools, such as image analysis, data management, process-based simulation, and upscaling tools, have been developed to help quantify and understand water flow in the soil-root system, at multiple scales (tissue, organ, plant, and population). Several of these tools work together or at least are compatible. However, for the uninformed researcher, they might seem disconnected, forming an unclear and disorganized succession of tools. In this article, we show how different studies can be further developed by connecting them to analyse soil-root water relations in a comprehensive and structured network. This 'explicit network of soil-root computational tools' informs readers about existing tools and helps them understand how their data (past and future) might fit within the network. We also demonstrate the novel possibilities of scale-consistent parameterizations made possible by the network with a set of case studies from the literature. Finally, we discuss existing gaps in the network and how we can move forward to fill them.

Water transport through tall trees: A vertically explicit, analytical model of xylem hydraulic conductance in stems.

Trees grow by vertically extending their stems, so accurate stem hydraulic models are fundamental to understanding the hydraulic challenges faced by tall trees. Using a literature survey, we showed that many tree species exhibit continuous vertical variation in hydraulic traits. To examine the effects of this variation on hydraulic function, we developed a spatially explicit, analytical water transport model for stems. Our model allows Huber ratio, stem-saturated conductivity, pressure at 50% loss of conductivity, leaf area, and transpiration rate to vary continuously along the hydraulic path. Predictions from our model differ from a matric flux potential model parameterized with uniform traits. Analyses show that cavitation is a whole-stem emergent property resulting from non-linear pressure-conductivity feedbacks that, with gravity, cause impaired water transport to accumulate along the path. Because of the compounding effects of vertical trait variation on hydraulic function, growing proportionally more sapwood and building tapered xylem with height, as well as reducing xylem vulnerability only at branch tips while maintaining transport capacity at the stem base, can compensate for these effects. We therefore conclude that the adaptive significance of vertical variation in stem hydraulic traits is to allow trees to grow tall and tolerate operating near their hydraulic limits.

Estimation of the hydraulic conductivities of lupine roots by inverse modelling of high-resolution measurements of root water uptake.

Background and Aims Radial and axial hydraulic conductivities are key parameters for proper understanding and modelling of root water uptake. Despite their importance, there is limited experimental information on how the radial and axial hydraulic conductivities vary along roots growing in soil. Here, a new approach was introduced to estimate inversely the profile of hydraulic conductivities along the roots of transpiring plants growing in soil. Methods A three-dimensional model of root water uptake was used to reproduce the measured profile of root water uptake along roots of lupine plant grown in soil. The profile of fluxes was measured using a neutron radiography technique combined with injection of deuterated water as tracer. The aim was to estimate inversely the profiles of the radial and axial hydraulic conductivities along the roots. Key Results The profile of hydraulic conductivities along the taproot and the lateral roots of lupines was calculated using three flexible scenarios. For all scenarios, it was found that the radial hydraulic conductivity increases towards the root tips, while the axial conductivity decreases. Additionally, it was found that in soil with uniform water content: (1) lateral roots were the main location of root water uptake; (2) water uptake by laterals decreased towards the root tips due to the dissipation of water potential along the root; and (3) water uptake by the taproot was higher in the distal segments and was negligible in the proximal parts, which had a low radial conductivity. Conclusions The proposed approach allows the estimation of the root hydraulic properties of plants growing in soil. This information can be used in an advanced model of water uptake to predict the water uptake of different root types or different root architectures under varying soil conditions.

Plant water uptake in drying soils.

Over the last decade, investigations on root water uptake have evolved toward a deeper integration of the soil and roots compartment properties, with the goal of improving our understanding of water acquisition from drying soils. This evolution parallels the increasing attention of agronomists to suboptimal crop production environments. Recent results have led to the description of root system architectures that might contribute to deep-water extraction or to water-saving strategies. In addition, the manipulation of root hydraulic properties would provide further opportunities to improve water uptake. However, modeling studies highlight the role of soil hydraulics in the control of water uptake in drying soil and call for integrative soil-plant system approaches.

Root hydraulic properties: An exploration of their variability across scales.

Root hydraulic properties are key physiological traits that determine the capacity of root systems to take up water, at a specific evaporative demand. They can strongly vary among species, cultivars or even within the same genotype, but a systematic analysis of their variation across plant functional types (PFTs) is still missing. Here, we reviewed published empirical studies on root hydraulic properties at the segment-, individual root-, or root system scale and determined its variability and the main factors contributing to it. This corresponded to a total of 241 published studies, comprising 213 species, including woody and herbaceous vegetation. We observed an extremely large range of variation (of orders of magnitude) in root hydraulic properties, but this was not caused by systematic differences among PFTs. Rather, the (combined) effect of factors such as root system age, driving force used for measurement, or stress treatments shaped the results. We found a significant decrease in root hydraulic properties under stress conditions (drought and aquaporin inhibition, p < .001) and a significant effect of the driving force used for measurement (hydrostatic or osmotic gradients, p < .001). Furthermore, whole root system conductance increased significantly with root system age across several crop species (p < .01), causing very large variation in the data (>2 orders of magnitude). Interestingly, this relationship showed an asymptotic shape, with a steep increase during the first days of growth and a flattening out at later stages of development. We confirmed this dynamic through simulations using a state-of-the-art computational model of water flow in the root system for a variety of crop species, suggesting common patterns across studies and species. These findings provide better understanding of the main causes of root hydraulic properties variations observed across empirical studies. They also open the door to better representation of hydraulic processes across multiple plant functional types and at large scales. All data collected in our analysis has been aggregated into an open access database (https://roothydraulic-properties.shinyapps.io/database/), fostering scientific exchange.

How to Define the Appropriate Spatial Resolution of Root Segments When Solving Water Flow in Root System Hydraulic Architectures.

In this chapter, we discuss the issue of balance between spatial resolution and computational efficiency in the context of the R-SWMS model. Based on the equations governing the water fluxes within the model, we propose here an objective and quantitative criterion which can help fix root segment size to both minimize computational load and achieve simulation according to a given accuracy degree.

Investigating Soil-Root Interactions with the Numerical Model R-SWMS.

In this chapter, we present the Root and Soil Water Movement and Solute transport model R-SWMS, which can be used to simulate flow and transport in the soil-plant system. The equations describing water flow in soil-root systems are presented and numerical solutions are provided. An application of R-SWMS is then briefly discussed, in which we combine in vivo and in silico experiments in order to decrypt water flow in the soil-root domain. More precisely, light transmission imaging experiments were conducted to generate data that can serve as input for the R-SWMS model. These data include the root system architecture, the soil hydraulic properties and the environmental conditions (initial soil water content and boundary conditions, BC). Root hydraulic properties were not acquired experimentally, but set to theoretical values found in the literature. In order to validate the results obtained by the model, the simulated and experimental water content distributions were compared. The model was then used to estimate variables that were not experimentally accessible, such as the actual root water uptake distribution and xylem water potential.

Hydraulic flux-responsive hormone redistribution determines root branching.

Plant roots exhibit plasticity in their branching patterns to forage efficiently for heterogeneously distributed resources, such as soil water. The xerobranching response represses lateral root formation when roots lose contact with water. Here, we show that xerobranching is regulated by radial movement of the phloem-derived hormone abscisic acid, which disrupts intercellular communication between inner and outer cell layers through plasmodesmata. Closure of these intercellular pores disrupts the inward movement of the hormone signal auxin, blocking lateral root branching. Once root tips regain contact with moisture, the abscisic acid response rapidly attenuates. Our study reveals how roots adapt their branching pattern to heterogeneous soil water conditions by linking changes in hydraulic flux with dynamic hormone redistribution.

Combining cross-section images and modeling tools to create high-resolution root system hydraulic atlases in Zea mays.

Root hydraulic properties play a central role in the global water cycle, in agricultural systems productivity, and in ecosystem survival as they impact the canopy water supply. However, the existing experimental methods to quantify root hydraulic conductivities, such as the root pressure probing, are particularly challenging, and their applicability to thin roots and small root segments is limited. Therefore, there is a gap in methods enabling easy estimations of root hydraulic conductivities in diverse root types. Here, we present a new pipeline to quickly estimate root hydraulic conductivities across different root types, at high resolution along root axes. Shortly, free-hand root cross-sections were used to extract a selected number of key anatomical traits. We used these traits to parametrize the Generator of Root Anatomy in R (GRANAR) model to simulate root anatomical networks. Finally, we used these generated anatomical networks within the Model of Explicit Cross-section Hydraulic Anatomy (MECHA) to compute an estimation of the root axial and radial hydraulic conductivities (k x and k r , respectively). Using this combination of anatomical data and computational models, we were able to create a root hydraulic conductivity atlas at the root system level, for 14-day-old pot-grown Zea mays (maize) plants of the var. B73. The altas highlights the significant functional variations along and between different root types. For instance, predicted variations of radial conductivity along the root axis were strongly dependent on the maturation stage of hydrophobic barriers. The same was also true for the maturation rates of the metaxylem vessels. Differences in anatomical traits along and across root types generated substantial variations in radial and axial conductivities estimated with our novel approach. Our methodological pipeline combines anatomical data and computational models to turn root cross-section images into a detailed hydraulic atlas. It is an inexpensive, fast, and easily applicable investigation tool for root hydraulics that complements existing complex experimental methods. It opens the way to high-throughput studies on the functional importance of root types in plant hydraulics, especially if combined with novel phenotyping techniques such as laser ablation tomography.

Non-invasive hydrodynamic imaging in plant roots at cellular resolution.

A key impediment to studying water-related mechanisms in plants is the inability to non-invasively image water fluxes in cells at high temporal and spatial resolution. Here, we report that Raman microspectroscopy, complemented by hydrodynamic modelling, can achieve this goal - monitoring hydrodynamics within living root tissues at cell- and sub-second-scale resolutions. Raman imaging of water-transporting xylem vessels in Arabidopsis thaliana mutant roots reveals faster xylem water transport in endodermal diffusion barrier mutants. Furthermore, transverse line scans across the root suggest water transported via the root xylem does not re-enter outer root tissues nor the surrounding soil when en-route to shoot tissues if endodermal diffusion barriers are intact, thereby separating 'two water worlds'.

Call for Participation: Collaborative Benchmarking of Functional-Structural Root Architecture Models. The Case of Root Water Uptake.

Three-dimensional models of root growth, architecture and function are becoming important tools that aid the design of agricultural management schemes and the selection of beneficial root traits. However, while benchmarking is common in many disciplines that use numerical models, such as natural and engineering sciences, functional-structural root architecture models have never been systematically compared. The following reasons might induce disagreement between the simulation results of different models: different representation of root growth, sink term of root water and solute uptake and representation of the rhizosphere. Presently, the extent of discrepancies is unknown, and a framework for quantitatively comparing functional-structural root architecture models is required. We propose, in a first step, to define benchmarking scenarios that test individual components of complex models: root architecture, water flow in soil and water flow in roots. While the latter two will focus mainly on comparing numerical aspects, the root architectural models have to be compared at a conceptual level as they generally differ in process representation. Therefore, defining common inputs that allow recreating reference root systems in all models will be a key challenge. In a second step, benchmarking scenarios for the coupled problems are defined. We expect that the results of step 1 will enable us to better interpret differences found in step 2. This benchmarking will result in a better understanding of the different models and contribute toward improving them. Improved models will allow us to simulate various scenarios with greater confidence and avoid bugs, numerical errors or conceptual misunderstandings. This work will set a standard for future model development.

Hydraulic conductivity of soil-grown lupine and maize unbranched roots and maize root-shoot junctions.

Improving or maintaining crop productivity under conditions of long term change of soil water availability and atmosphere demand for water is one the big challenges of this century. It requires a deep understanding of crop water acquisition properties, i.e. root system architecture and root hydraulic properties among other characteristics of the soil-plant-atmosphere continuum. A root pressure probe technique was used to measure the root hydraulic conductances of seven-week old maize and lupine plants grown in sandy soil. Unbranched root segments were excised in lateral, seminal, crown and brace roots of maize, and in lateral roots of lupine. Their total hydraulic conductance was quantified under steady-state hydrostatic gradient for progressively shorter segments. Furthermore, the axial conductance of proximal root regions removed at each step of root shortening was measured as well. Analytical solutions of the water flow equations in unbranched roots developed recently and relating root total conductance profiles to axial and radial conductivities were used to retrieve the root radial hydraulic conductivity profile along each root type, and quantify its uncertainty. Interestingly, the optimized root radial conductivities and measured axial conductances displayed significant differences across root types and species. However, the measured root total conductances did not differ significantly. As compared to measurements reported in the literature, our axial and radial conductivities concentrate in the lower range of herbaceous species hydraulic properties. In a final experiment, the hydraulic conductances of root junctions to maize stem were observed to highly depend on root type. Surprisingly maize brace root junctions were an order of magnitude more conductive than the other crown and seminal roots, suggesting potential regulation mechanism for root water uptake location and a potential role of the maize brace roots for water uptake more important than reported in the literature.