The nematode effector Mj-NEROSs interacts with Rieske's iron-sulfur protein influencing plastid ROS production to suppress plant immunity.

Root-knot nematodes (RKN; Meloidogyne species) are plant pathogens that introduce several effectors in their hosts to facilitate infection. The actual targets and functioning mechanism of these effectors largely remain unexplored. This study illuminates the role and interplay of the Meloidogyne javanica nematode effector ROS suppressor (Mj-NEROSs) within the host plant environment. Mj-NEROSs suppresses INF1-induced cell death as well as flg22-induced callose deposition and reactive oxygen species (ROS) production. A transcriptome analysis highlighted the downregulation of ROS-related genes upon Mj-NEROSs expression. NEROSs interacts with the plant Rieske's iron-sulfur protein (ISP) as shown by yeast-two-hybrid and bimolecular fluorescence complementation. Secreted from the subventral pharyngeal glands into giant cells, Mj-NEROSs localizes in the plastids where it interacts with ISP, subsequently altering electron transport rates and ROS production. Moreover, our results demonstrate that isp Arabidopsis thaliana mutants exhibit increased susceptibility to M. javanica, indicating ISP importance for plant immunity. The interaction of a nematode effector with a plastid protein highlights the possible role of root plastids in plant defense, prompting many questions on the details of this process.

Mechanistic modeling reveals the importance of turgor-driven apoplastic water transport in wheat stem parenchyma during carbohydrate mobilization.

During stem elongation, wheat (Triticum aestivum) increases its stem carbohydrate content before anthesis as a reserve for grain filling. Hydraulic functioning during this mobilization process is not well understood, and contradictory results exist on the direct effect of drought on carbohydrate mobilization. In a dedicated experiment, wheat plants were subjected to drought stress during carbohydrate mobilization. Measurements, important to better understand stem physiology, showed some unexpected patterns that could not be explained by our current knowledge on water transport. Traditional water flow and storage models failed to properly describe the drought response in wheat stems during carbohydrate mobilization. To explain the measured patterns, hypotheses were formulated and integrated in a dedicated model for wheat. The new mechanistic model simulates two hypothetical water storage compartments: one where water is quickly exchanged with the xylem and one that contains the carbohydrate storage. Water exchange between these compartments is turgor-driven. The model was able to simulate the measured increase in stored carbohydrate concentrations with a decrease in water content and stem diameter. Calibration of the model showed the importance of turgor-driven apoplastic water flow during carbohydrate mobilization. This resulted in an increase in stem hydraulic capacitance, which became more important under drought stress.

Daytime stomatal regulation in mature temperate trees prioritizes stem rehydration at night.

Trees remain sufficiently hydrated during drought by closing stomata and reducing canopy conductance (Gc ) in response to variations in atmospheric water demand and soil water availability. Thresholds that control the reduction of Gc are proposed to optimize hydraulic safety against carbon assimilation efficiency. However, the link between Gc and the ability of stem tissues to rehydrate at night remains unclear. We investigated whether species-specific Gc responses aim to prevent branch embolisms, or enable night-time stem rehydration, which is critical for turgor-dependent growth. For this, we used a unique combination of concurrent dendrometer, sap flow and leaf water potential measurements and collected branch-vulnerability curves of six common European tree species. Species-specific Gc reduction was weakly related to the water potentials at which 50% of branch xylem conductivity is lost (P50 ). Instead, we found a stronger relationship with stem rehydration. Species with a stronger Gc control were less effective at refilling stem-water storage as the soil dries, which appeared related to their xylem architecture. Our findings highlight the importance of stem rehydration for water-use regulation in mature trees, which likely relates to the maintenance of adequate stem turgor. We thus conclude that stem rehydration must complement the widely accepted safety-efficiency stomatal control paradigm.

Limited plasticity of anatomical and hydraulic traits in aspen trees under elevated CO2 and seasonal drought.

The timing of abiotic stress elicitors on wood formation largely affects xylem traits that determine xylem efficiency and vulnerability. Nonetheless, seasonal variability of elevated CO2 (eCO2) effects on tree functioning under drought remains largely unknown. To address this knowledge gap, 1-year-old aspen (Populus tremula L.) trees were grown under ambient (±445 ppm) and elevated (±700 ppm) CO2 and exposed to an early (spring/summer 2019) or late (summer/autumn 2018) season drought event. Stomatal conductance and stem shrinkage were monitored in vivo as xylem water potential decreased. Additional trees were harvested for characterization of wood anatomical traits and to determine vulnerability and desorption curves via bench dehydration. The abundance of narrow vessels decreased under eCO2 only during the early season. At this time, xylem vulnerability to embolism formation and hydraulic capacitance during severe drought increased under eCO2. Contrastingly, stomatal closure was delayed during the late season, while hydraulic vulnerability and capacitance remained unaffected under eCO2. Independently of the CO2 treatment, elastic, and inelastic water pools depleted simultaneously after 50% of complete stomatal closure. Our results suggest that the effect of eCO2 on drought physiology and wood traits are small and variable during the growing season and question a sequential capacitive water release from elastic and inelastic pools as drought proceeds.

High vapour pressure deficit enhances turgor limitation of stem growth in an Asian tropical rainforest tree.

Tropical forests are experiencing increases in vapour pressure deficit (D), with possible negative impacts on tree growth. Tree-growth reduction due to rising D is commonly attributed to carbon limitation, thus overlooking the potentially important mechanism of D-induced impairment of wood formation due to an increase in turgor limitation. Here we calibrate a mechanistic tree-growth model to simulate turgor limitation of radial stem growth in mature Toona cilitata trees in an Asian tropical forest. Hourly sap flow and dendrometer measurements were collected to simulate turgor-driven growth during the growing season. Simulated seasonal patterns of radial stem growth matched well with growth observations. Growth mainly occurred at night and its pre-dawn build-up appeared to be limited under higher D. Across seasons, the night-time turgor pressure required for growth was negatively related to previous midday D, possibly due to a relatively high canopy conductance at high D, relative to stem rehydration. These findings provide the first evidence that tropical trees grow at night and that turgor pressure limits tree growth. We suggest including turgor limitation of tree stem growth in models also for tropical forest carbon dynamics, in particular, if these models simulate effects of warming and increased frequency of droughts.

Differences between tree stem CO2 efflux and O2 influx rates cannot be explained by internal CO2 transport or storage in large beech trees.

Tree stem respiration (RS ) is a substantial component of the forest carbon balance. The mass balance approach uses stem CO2 efflux and internal xylem fluxes to sum up RS , while the oxygen-based method assumes O2 influx as a proxy of RS . So far, both approaches have yielded inconsistent results regarding the fate of respired CO2 in tree stems, a major challenge for quantifying forest carbon dynamics. We collected a data set of CO2 efflux, O2 influx, xylem CO2 concentration, sap flow, sap pH, stem temperature, nonstructural carbohydrates concentration and potential phosphoenolpyruvate carboxylase (PEPC) capacity on mature beech trees to identify the sources of differences between approaches. The ratio of CO2 efflux to O2 influx was consistently below unity (0.7) along a 3-m vertical gradient, but internal fluxes did not bridge the gap between influx and efflux, nor did we find evidence for changes in respiratory substrate use. PEPC capacity was comparable with that previously reported in green current-year twigs. Although we could not reconcile differences between approaches, results shed light on the uncertain fate of CO2 respired by parenchyma cells across the sapwood. Unexpected high values of PEPC capacity highlight its potential relevance as a mechanism of local CO2 removal, which merits further research.

Climatic niche lability but growth form conservatism in the African woody flora.

Climatic niche evolution during the diversification of tropical plants has received little attention in Africa. To address this, we characterised the climatic niche of >4000 tropical African woody species, distinguishing two broad bioclimatic groups (forest vs. savanna) and six subgroups. We quantified niche conservatism versus lability at the genus level and for higher clades, using a molecular phylogeny of >800 genera. Although niche stasis at speciation is prevalent, numerous clades individually cover vast climatic spaces suggesting a general ease in transcending ecological limits, especially across bioclimatic subgroups. The forest biome was the main source of diversity, providing many lineages to savanna, but reverse shifts also occurred. We identified clades that diversified in savanna after shifts from forest. The forest-savanna transition was not consistently associated with a growth form change, though we found evolutionarily labile clades whose presence in forest or savanna is associated respectively with climbing or shrubby species diversification.

Mechanistic drivers of stem respiration: A modelling exercise across species and seasons.

Stem respiration (RS ) plays a crucial role in plant carbon budgets. However, its poor understanding limits our ability to model woody tissue and whole-tree respiration. A biophysical model of stem water and carbon fluxes (TReSpire) was calibrated on cedar, maple and oak trees during spring and late summer. For this, stem sap flow, water potential, diameter variation, temperature, CO2 efflux, allometry and biochemistry were monitored. Shoot photosynthesis (PN ) and nonstructural carbohydrates (NSC) were additionally measured to evaluate source-sink relations. The highest RS and stem growth was found in maple and oak during spring, both being seasonally decoupled from PN and [NSC]. Temperature largely affected maintenance respiration (RM ) in the short term, but temperature-normalized RM was highly variable on a seasonal timescale. Overall, most of the respired CO2 radially diffused to the atmosphere (>87%) while the remainder was transported upward with the transpiration stream. The modelling exercise highlights the sink-driven behaviour of RS and the significance of overall metabolic activity on nitrogen (N) allocation patterns and N-normalized respiratory costs to capture RS variability over the long term. These insights should be considered when modelling plant respiration, whose representation is currently biased towards a better understanding of leaf metabolism.

Foliar water uptake does not contribute to embolism repair in beech (Fagus sylvatica L.).

BACKGROUND AND AIMS: Foliar water uptake has recently been suggested as a possible mechanism for the restoration of hydraulically dysfunctional xylem vessels. In this paper we used a combination of ecophysiological measurements, X-ray microcomputed tomography and cryo-scanning electron microscopy during a drought treatment to fully evaluate this hypothesis. KEY RESULTS: Based on an assessment of these methods in beech (Fagus sylvatica L.) seedlings we were able to (1) confirm an increase in the amount of hydraulically redistributed water absorbed by leaves when the soil water potential decreased, and (2) locate this redistributed water in hydraulically active vessels in the stem. However, (3) no embolism repair was observed irrespective of the organ under investigation (i.e. stem, petiole or leaf) or the intensity of drought. CONCLUSIONS: Our data provide evidence for a hydraulic pathway from the leaf surface to the stem xylem following a water potential gradient, but this pathway exists only in functional vessels and does not play a role in embolism repair for beech.

Identifying the pathways for foliar water uptake in beech (Fagus sylvatica L.): a major role for trichomes.

Foliar water uptake (FWU), the direct uptake of water into leaves, is a global phenomenon, having been observed in an increasing number of plant species. Despite the growing recognition of its functional relevance, our understanding of how FWU occurs and which foliar surface structures are implicated, is limited. In the present study, fluorescent and ionic tracers, as well as microcomputed tomography, were used to assess potential pathways for water entry in leaves of beech, a widely distributed tree species from European temperate regions. Although none of the tracers entered the leaf through the stomatal pores, small amounts of silver precipitation were observed in some epidermal cells, indicating moderate cuticular uptake. Trichomes, however, were shown to absorb and redistribute considerable amounts of ionic and fluorescent tracers. Moreover, microcomputed tomography indicated that 72% of empty trichomes refilled during leaf surface wetting and microscopic investigations revealed that trichomes do not have a cuticle but are covered with a pectin-rich cell wall layer. Taken together, our findings demonstrate that foliar trichomes, which exhibit strong hygroscopic properties as a result of their structural and chemical design, constitute a major FWU pathway in beech.

Turgor - a limiting factor for radial growth in mature conifers along an elevational gradient.

A valid representation of intra-annual wood formation processes in global vegetation models is vital for assessing climate change impacts on the forest carbon stock. Yet, wood formation is generally modelled with photosynthesis, despite mounting evidence that cambial activity is rather directly constrained by limiting environmental factors. Here, we apply a state-of-the-art turgor-driven growth model to simulate 4 yr of hourly stem radial increment from Picea abies (L.) Karst. and Larix decidua Mill. growing along an elevational gradient. For the first time, wood formation observations were used to validate weekly to annual stem radial increment simulations, while environmental measurements were used to assess the climatic constraints on turgor-driven growth. Model simulations matched the observed timing and dynamics of wood formation. Using the detailed model outputs, we identified a strict environmental regulation on stem growth (air temperature > 2°C and soil water potential > -0.6 MPa). Warmer and drier summers reduced the growth rate as a result of turgor limitation despite warmer temperatures being favourable for cambial activity. These findings suggest that turgor is a central driver of the forest carbon sink and should be considered in next-generation vegetation models, particularly in the context of global warming and increasing frequency of droughts.

Temporal variability in tree responses to elevated atmospheric CO2.

At leaf level, elevated atmospheric CO2 concentration (eCO2 ) results in stimulation of carbon net assimilation and reduction of stomatal conductance. However, a comprehensive understanding of the impact of eCO2 at larger temporal (seasonal and annual) and spatial (from leaf to whole-tree) scales is still lacking. Here, we review overall trends, magnitude and drivers of dynamic tree responses to eCO2 , including carbon and water relations at the leaf and the whole-tree level. Spring and early season leaf responses are most susceptible to eCO2 and are followed by a down-regulation towards the onset of autumn. At the whole-tree level, CO2 fertilization causes consistent biomass increments in young seedlings only, whereas mature trees show a variable response. Elevated CO2 -induced reductions in leaf stomatal conductance do not systematically translate into limitation of whole-tree transpiration due to the unpredictable response of canopy area. Reduction in the end-of-season carbon sink demand and water-limiting strategies are considered the main drivers of seasonal tree responses to eCO2 . These large temporal and spatial variabilities in tree responses to eCO2 highlight the risk of predicting tree behavior to eCO2 based on single leaf-level point measurements as they only reveal snapshots of the dynamic responses to eCO2 .

Efflux and assimilation of xylem-transported CO2 in stems and leaves of tree species with different wood anatomy.

Determining the fate of CO2 respired in woody tissues is necessary to understand plant respiratory physiology and to evaluate CO2 recycling mechanisms. An aqueous 13 C-enriched CO2 solution was infused into the stem of 3-4 m tall trees to estimate efflux and assimilation of xylem-transported CO2 via cavity ring-down laser spectroscopy and isotope ratio mass spectrometry, respectively. Different tree locations (lower stem, upper stem and leafy shoots) and tissues (xylem, bark and leaves) were monitored in species with tracheid, diffuse- and ring-porous wood anatomy (cedar, maple and oak, respectively). Radial xylem CO2 diffusivity and xylem [CO2 ] were lower in cedar relative to maple and oak trees, thereby limiting label diffusion. Part of the labeled 13 CO2 was assimilated in cedar (8.7%) and oak (20.6%) trees, mostly in xylem and bark tissues of the stem, while limited solution uptake in maple trees hindered the detection of label assimilation. Little label reached foliar tissues, suggesting substantial label loss along the stem-branch transition following reductions in the radial diffusive pathway. Differences in respiration rates and radial xylem CO2 diffusivity (lower in conifer relative to angiosperm species) might reconcile discrepancies in efflux and assimilation of xylem-transported CO2 so far observed between taxonomic clades.

Turgor-time controls grass leaf elongation rate and duration under drought stress.

The process of leaf elongation in grasses is characterized by the creation and transformation of distinct cell zones. The prevailing turgor pressure within these cells is one of the key drivers for the rate at which these cells divide, expand and differentiate, processes that are heavily impacted by drought stress. In this article, a turgor-driven growth model for grass leaf elongation is presented, which combines mechanistic growth from the basis of turgor pressure with the ontogeny of the leaf. Drought-induced reductions in leaf turgor pressure result in a simultaneous inhibition of both cell expansion and differentiation, lowering elongation rate but increasing elongation duration due to the slower transitioning of cells from the dividing and elongating zone to mature cells. Leaf elongation is, therefore, governed by the magnitude of, and time spent under, growth-enabling turgor pressure, a metric which we introduce as turgor-time. Turgor-time is able to normalize growth patterns in terms of varying water availability, similar to how thermal time is used to do so under varying temperatures. Moreover, additional inclusion of temperature dependencies within our model pioneers a novel concept enabling the general expression of growth regardless of water availability or temperature.

Woody tissue photosynthesis delays drought stress in Populus tremula trees and maintains starch reserves in branch xylem tissues.

Photosynthesis in woody tissues (Pwt ) is less sensitive to water shortage than in leaves, hence, Pwt might be a crucial carbon source to alleviate drought stress. To evaluate the impact of Pwt on tree drought tolerance, woody tissues of 4-m-tall drought-stressed Populus tremula trees were subjected to a light-exclusion treatment across the entire plant to inhibit Pwt . Xylem water potential (Ψxylem ), sap flow ( FH2O ), leaf net photosynthesis (Pn,l ), stem diameter variations (ΔD), in vivo acoustic emissions in stems (AEs) and nonstructural carbohydrate concentrations ([NSC]) were monitored to comprehensively assess water and carbon relations at whole-tree level. Under well-watered conditions, Pwt kept Ψxylem at a higher level, lowered FH2O and had no effect on [NSC]. Under drought, Ψxylem , FH2O and Pn,l in light-excluded trees rapidly decreased in concert with reductions in branch xylem starch concentration. Moreover, sub-daily patterns of ΔD, FH2O and AEs were strongly related, suggesting that in vivo AEs may inform not only about embolism events, but also about capacitive release and replenishment of stem water pools. Results highlight the importance of Pwt in maintaining xylem hydraulic integrity under drought conditions and in sustaining NSC pools to potentially limit increases in xylem tension.

TReSpire - a biophysical TRee Stem respiration model.

Mechanistic models of plant respiration remain poorly developed, especially in stems and woody tissues where measurements of CO2 efflux do not necessarily reflect local respiratory activity. We built a process-based model of stem respiration that couples water and carbon fluxes at the organ level (TReSpire). To this end, sap flow, stem diameter variations, xylem and soil water potential, stem temperature, stem CO2 efflux and nonstructural carbohydrates were measured in a maple tree, while xylem CO2 concentration and additional stem and xylem diameter variations were monitored in an ancillary tree for model validation. TReSpire realistically described: (1) turgor pressure to differentiate growing from nongrowing metabolism; (2) maintenance expenditures in xylem and outer tissues based on Arrhenius kinetics and nitrogen content; and (3) radial CO2 diffusivity and CO2 solubility and transport in the sap solution. Collinearity issues with phloem unloading rates and sugar-starch interconversion rates suggest parallel submodelling to close the stem carbon balance. TReSpire brings a breakthrough in the modelling of stem water and carbon fluxes at a detailed (hourly) temporal resolution. TReSpire is calibrated from a sink-driven perspective, and has potential to advance our understanding on stem growth dynamics, CO2 fluxes and underlying respiratory physiology across different species and phenological stages.

Woody tissue photosynthesis reduces stem CO2 efflux by half and remains unaffected by drought stress in young Populus tremula trees.

A substantial portion of locally respired CO2 in stems can be assimilated by chloroplast-containing tissues. Woody tissue photosynthesis (Pwt ) therefore plays a major role in the stem carbon balance. To study the impact of Pwt on stem carbon cycling along a gradient of water availability, stem CO2 efflux (EA ), xylem CO2 concentration ([CO2 ]), and xylem water potential (Ψxylem ) were measured in 4-year-old Populus tremula L. trees exposed to drought stress and different regimes of light exclusion of woody tissues. Under well-watered conditions, local Pwt decreased EA up to 30%. Axial CO2 diffusion (Dax ) induced by distant Pwt caused an additional decrease in EA of up to 25% and limited xylem [CO2 ] build-up. Under drought stress, absolute decreases in EA driven by Pwt remained stable, denoting that Pwt was not affected by drought. At the end of the dry period, when transpiration was low, local Pwt and Dax offset 20% and 10% of stem respiration on a daily basis, respectively. These results highlight (a) the importance of Pwt for an adequate interpretation of EA measurements and (b) homeostatic Pwt along a drought stress gradient, which might play a crucial role to fuel stem metabolism when leaf carbon uptake and phloem transport are limited.

Hydraulic acclimation in a Mediterranean oak subjected to permanent throughfall exclusion results in increased stem hydraulic capacitance.

Stem water storage capacity and hydraulic capacitance (CS ) play a crucial role in tree survival under drought-stress. To investigate whether CS adjusts to increasing water deficit, variation in stem water content (StWC) was monitored in vivo for 2 years and related to periodical measurements of tree water potential in Mediterranean Quercus ilex trees subjected either to permanent throughfall exclusion (TE) or to control conditions. Seasonal reductions in StWC were larger in TE trees relative to control ones, resulting in greater seasonal CS (154 and 80 kg m-3 MPa-1 , respectively), but only during the first phase of the desorption curve, when predawn water potential was above -1.1 MPa. Below this point, CS decreased substantially and did not differ between treatments (<20 kg m-3 MPa-1 ). The allometric relationship between tree diameter and sapwood area, measured via electrical resistivity tomography, was not affected by TE. Our results suggest that (a) CS response to water deficit in the drought-tolerant Q. ilex might be more important to optimize carbon gain during well-hydrated periods than to prevent drought-induced embolism formation during severe drought stress, and (b) enhanced CS during early summer does not result from proportional increases in sapwood volume, but mostly from increased elastic water.

Turgor-driven plant growth applied in a soybean functional-structural plant model.

BACKGROUND AND AIMS: Turgor pressure within a plant cell represents the key to the mechanistical descriptiion of plant growth, combining the effects of both water and carbon availability. The high level of spatio-temporal variation and diurnal dynamics in turgor pressure within a single plant make it a challenge to model these on the fine spatial scale required for functional-structural plant models (FSPMs). A conceptual model for turgor-driven growth in FSPMs has been established previously, but its practical use has not yet been explored. METHODS: A turgor-driven growth model was incorporated in a newly established FSPM for soybean. The FSPM simulates dynamics in photosynthesis, transpiration and turgor pressure in direct relation to plant growth. Comparisons of simulations with field data were used to evaluate the potential and shortcomings of the modelling approach. KEY RESULTS: Model simulations revealed the need to include an initial seed carbon contribution, a more realistic sink function, an estimation of respiration, and the distinction between osmotic and structural sugars, in order to achieve a realistic model of plant growth. However, differences between simulations and observations remained in individual organ growth patterns and under different environmental conditions. This exposed the need to further investigate the assumptions of developmental and environmental (in)sensitivity of the parameters, which represent physiological and biophysical organ properties in the model, in future research. CONCLUSIONS: The model in its current form is primarily a diagnostic tool, to better understand and model the behaviour of water relations on the scale of individual plant organs throughout the plant life cycle. Potential future applications include its use as a phenotyping tool to capture differences in plant performance between genotypes and growing environments in terms of specific plant characteristics. Additionally, focused experiments can be used to further improve the model mechanisms to lead to better predictive FSPMs, including scenarios of water deficit.

Quantifying the importance of a realistic tomato (Solanum lycopersicum) leaflet shape for 3-D light modelling.

BACKGROUND AND AIMS: Leaflet shapes of tomato plants (Solanum lycopersicum) have been reduced to simple geometric shapes in previous functional-structural plant models (FSPMs) in order to facilitate measurements and reduce the time required to reconstruct the plant virtually. The level of error that such simplifications introduce remains unaddressed. This study therefore aims to quantify the modelling error associated with simplifying leaflet shapes. METHODS: Realistic shapes were implemented in a static tomato FSPM based on leaflet scans, and simulation results were compared to simple geometric shapes used in previous tomato FSPMs in terms of light absorption and gross photosynthesis, for both a single plant and a glasshouse scenario. KEY RESULTS: The effect of simplifying leaflet shapes in FSPMs leads to small but significant differences in light absorption, alterations of canopy light conditions and differences in photosynthesis. The magnitude of these differences depends on both the type of leaflet shape simplification used and the canopy shape and density. Incorporation of realistic shapes requires a small increase in initial measurement and modelling work to establish a shape database and comes at the cost of a slight increase in computation time. CONCLUSIONS: Our findings indicate that the error associated with leaflet shape simplification is small, but often unpredictable, and is affected by plant structure but also lamp placement, which is often a primary optimization goal of these static models. Assessment of the cost-benefit of realistic shape inclusion shows relatively little drawbacks for a decrease in model uncertainty.