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Background Plants have adapted to survive seasonal life-threatening frost and drought. However, the timing and frequency of such events are impacted by climate change, jeopardizing plant survival. Understanding better the strategies of survival to dehydration stress is therefore timely and can be enhanced by the cross-fertilization of research between disciplines (ecology, physiology), models (woody, herbaceous species) and types of stress (drought, frost). Scope We build upon the ‘growth–stress survival’ trade-off, which underpins the identification of global plant strategies across environments along a ‘fast–slow’ economics spectrum. Although phenological adaptations such as dormancy are crucial to survive stress, plant global strategies along the fast–slow economic spectrum rarely integrate growth variations across seasons. We argue that the growth–stress survival trade-off can be a useful framework to identify convergent plant ecophysiological strategies to survive both frost and drought. We review evidence that reduced physiological activity, embolism resistance and dehydration tolerance of meristematic tissues are interdependent strategies that determine thresholds of mortality among plants under severe frost and drought. We show that complete dormancy, i.e. programmed growth cessation, before stress occurrence, minimizes water flows and maximizes dehydration tolerance during seasonal life-threatening stresses. We propose that incomplete dormancy, i.e. the programmed reduction of growth potential during the harshest seasons, could be an overlooked but major adaptation across plants. Quantifying stress survival in a range of non-dormant versus winter- or summer-dormant plants, should reveal to what extent incomplete to complete dormancy could represent a proxy for dehydration tolerance and stress survival. Conclusions Our review of the strategies involved in dehydration stress survival suggests that winter and summer dormancy are insufficiently acknowledged as plant ecological strategies. Incorporating a seasonal fast–slow economics spectrum into global plant strategies improves our understanding of plant resilience to seasonal stress and refines our prevision of plant adaptation to extreme climatic events.

The high-capacity tensiometer and thermocouple psychrometer measure xylem water potential with high accuracy and adequate response time, as demonstrated by simultaneous measurements on sapling stems., The pressure chamber, the most popular method used to measure xylem water potential, is a discontinuous and destructive technique and is therefore not suitable for automated monitoring. Continuous non-destructive monitoring could until very recently be achieved only by use of the thermocouple psychrometer (TP). Here we present the high-capacity tensiometer (HCT) as an alternative method for continuous non-destructive monitoring. This provided us with a unique chance to cross-validate the two instruments by installing them simultaneously on the same sapling stem. The HCT and the TP showed excellent agreement for xylem water potential less than –0.5 MPa. Response to day/night cycles and watering was remarkably in phase, indicating excellent response time of both instruments despite substantially different working principles. For xylem water potential greater than –0.5 MPa, the discrepancies sometimes observed between the HCT and TP were mainly attributed to the kaolin paste used to establish contact between the xylem and the HCT, which becomes hydraulically poorly conductive in this range of water potential once dried beyond its air-entry value and subsequently re-wetted. Notwithstanding this limitation, which can be overcome by selecting a clay paste with higher air-entry value, the HCT has been shown to represent a valid alternative to the TP.

Climate change is challenging the resilience of grapevine (Vitis), one of the most important crops worldwide. Adapting viticulture to a hotter and drier future will require a multifaceted approach including the breeding of more drought-tolerant genotypes. In this study, we focused on plant hydraulics as a multi-trait system that allows the plant to maintain hydraulic integrity and gas exchange rates longer under drought. We quantified a broad range of drought-related traits within and across Vitis species, created in silico libraries of trait combinations, and then identified drought tolerant trait syndromes. By modeling the maintenance of hydraulic integrity of current cultivars and the drought tolerant trait syndromes, we identified elite ideotypes that increased the amount of time they could experience drought without leaf hydraulic failure. Generally, elites exhibited a trait syndrome with lower stomatal conductance, earlier stomatal closure, and a larger hydraulic safety margin. We demonstrated that, when compared with current cultivars, elite ideotypes have the potential to decrease the risk of hydraulic failure across wine regions under future climate scenarios. This study reveals the syndrome of traits that can be leveraged to protect grapevine from experiencing hydraulic failure under drought and increase drought tolerance.

We report the design and evaluation of pH responsive luminescent europium(III) probes that allow conjugation to targeting vectors to monitor receptor internalisation in cells. The approach adopted here can be used to tag proteins selectively and to monitor uptake into more acidic organelles, thereby enhancing the performance of time-resolved internalisation assays that require pH monitoring in real time.

A set of four luminescent EuIII complexes bearing an extended aryl‐alkynylpyridine chromophore has been studied, showing very different pH‐dependent behaviour in their absorption and emission spectral response. For two complexes with pK a values of 6.45 and 6.20 in protein‐containing solution, the emission lifetime increases very significantly following protonation. By varying the gate time during signal acquisition, the ‘switch‐on’ intensity ratio could be optimised, and enhancement factors of between 250 to 1330 were measured between pH 8 and 4. The best‐behaved probe showed no significant emission dependence on the concentration of endogenous cations, reductants, and serum albumin. It was examined in live‐cell imaging studies to monitor time‐dependent lysosomal acidification, for which the increase in observed image brightness due to acidification was a factor of 50 in NIH‐3T3 cells., Watching lysosomes age with a ‘switch‐on’ optical sensor is shown to be possible by using four europium complexes with extended aryl‐alkylnyl pyridine antennae that show very different pH‐dependent behaviour. A ‘switch‐on’ of europium luminescence occurs at low pH, allowing confocal microscopy to reveal increases in image brightness of ageing lysosomes in live‐cell imaging studies.

International audience; Lycophytes are the earliest diverging extant lineage of vascular plants, sister to all other vas-cular plants. Given that most species are adapted to ever-wet environments, it has been hypothesized that lycophytes, and by extension the common ancestor of all vascular plants, have few adaptations to drought. We investigated the responses to drought of key fitness-related traits such as stomatal regulation , shoot hydraulic conductance (K shoot) and stem xylem embolism resistance in Selaginella haematodes and S. pulcherrima, both native to tropical understory. During drought stomata in both species were found to close before declines in K shoot , with a 50% loss of K shoot occurring at À1.7 and À2.5 MPa in S. haematodes and S. pulcherrima, respectively. Direct observational methods revealed that the xylem of both species was resistant to embolism formation, with 50% of embolized xylem area occurring at À3.0 and À4.6 MPa in S. haematodes and S. pulcherrima, respectively. X-ray microcomputed tomogra-phy images of stems revealed that the decline in K shoot occurred with the formation of an air-filled lacuna, disconnecting the central vascular cylinder from the cortex. We propose that embolism-resistant xylem and large capacitance, provided by collapsing inner cortical cells, is essential for Selaginella survival during water deficit.

International audience; Identifying the drivers of stomatal closure and leaf damage during stress in grasses is a critical prerequisite for understanding crop resilience. Here, we investigated whether changes in stomatal conductance (g(s)) during dehydration were associated with changes in leaf hydraulic conductance (K-leaf), xylem cavitation, xylem collapse, and leaf cell turgor in wheat (Triticum aestivum). During soil dehydration, the decline of g(s) was concomitant with declining K-leaf under mild water stress. This early decline of leaf hydraulic conductance was not driven by cavitation, as the first cavitation events in leaf and stem were detected well after K-leaf had declined. Xylem vessel deformation could only account for

Context Understanding plant resilience and adaptation to drought is a major challenge in crop and forest sciences. Several methods have been developed to assess the vulnerability to xylem embolism. The in situ flow centrifuge (or cavitron) is the fastest technique allowing to characterise this trait for plants having vessel lengths shorter than the rotor size. Aims We present (i) a series of changes to the earlier cavitron design, aimed at improving the accuracy and speed of measurement through automated operations, and (ii) a new development through the design of a large diameter rotor expanding the range of species that can be measured. Methods Both hardware and software modifications to the original design have been developed. In order to avoid artefacts caused by cut open vessels, a centrifuge with a large rotor (1 m) has been developed, and vulnerability curves obtained with this new device were compared with those obtained using reference methods. Results The new set-up expands the range of conductance measurable with a cavitron and enables it to accurately determine the absolute value of conductivity even for species having very low hydraulic conductivity. The large rotor cavitron shows good agreement with the reference techniques for conifers and diffuse-porous species but also for ring-porous species having long vessels. Conclusion The set-up described in this manuscript provides a faster, safer and more accurate method to construct vulnerability curves, compared to the original cavitron design, and extends the measurement capabilities to new species that are difficult to measure to date. Key message Recent improvements to cavitron setup enable to measure xylem vulnerability curves for an expanded number of plant species, with longer vessels or lower hydraulic conductivity.

Xylem anatomy may change in response to environmental or biotic stresses. Vascular occlusion, an anatomical modification of mature xylem, contributes to plant resistance and susceptibility to different stresses. In woody organs, xylem occlusions have been examined as part of the senescence process, but their presence and function in leaves remain obscure. In grapevine, many stresses are associated with premature leaf senescence inducing discolorations and scorched tissue in leaves. However, we still do not know whether the leaf senescence process follows the same sequence of physiological events and whether leaf xylem anatomy is affected in similar ways. In this study, we quantified vascular occlusions in midribs from leaves with symptoms of the grapevine disease esca, magnesium deficiency and autumn senescence. We found higher amounts of vascular occlusions in leaves with esca symptoms (in 27% of xylem vessels on average), whereas the leaves with other symptoms (as well as the asymptomatic controls) had far fewer occlusions (in 3% of vessels). Therefore, we assessed the relationship between xylem occlusions and esca leaf symptoms in four different countries (California in the USA, France, Italy and Spain) and eight different cultivars. We monitored the plants over the course of the growing season, confirming that vascular occlusions do not evolve with symptom age. Finally, we investigated the hydraulic integrity of leaf xylem vessels by optical visualization of embolism propagation during dehydration. We found that the occlusions lead to hydraulic dysfunction mainly in the peripheral veins compared with the midribs in esca symptomatic leaves. These results open new perspectives on the role of vascular occlusions during the leaf senescence process, highlighting the uniqueness of esca leaf symptoms and its consequence on leaf physiology.

International audience; 1. Xylem hydraulic safety and efficiency are key traits determining tree fitness in a warmer and drier world. While numerous plant hydraulic studies have focused on branches, our understanding of root hydraulic functioning remains limited, although roots control water uptake, influence stomatal regulation and have commonly been considered as the most vulnerable organ along the hydraulic pathway.2. We investigated 11 traits related to xylem safety and efficiency along the hydraulic pathway in four temperate broad-leaved tree species.3. Continuous vessel tapering from coarse roots to stems and branches caused considerable reduction in hydraulic efficiency. Wood density was always lowest in roots, but did not decline linearly along the flow path. In contrast, xylem embolism resistance (P-50) did not differ significantly between roots and branches, except for one species. The limited variation in xylem safety between organs did not adequately reflect the corresponding reductions in vessel diameter (by similar to 70%) and hydraulic efficiency (by similar to 85%). Although we did not observe any trade-off between xylem safety and specific conductivity, vessel diameter, vessel lumen fraction and wood density were related to embolism resistance, both across and partly within organs.4. We conclude that coarse roots are not highly vulnerable to xylem embolism as commonly believed, indicating that hydraulic failure during soil drying might be restricted to fine roots. A free Plain Language Summary can be found within the Supporting Information of this article.

European beech is one of the most common tree species in Europe and is generally suggested to play even more of a prominent role in forestry in the future. It seems to have the potential to partially replace Norway spruce, as it is less sensitive to expected warmer and drier conditions. It is, however, not well known in which regions these new plantings would be particularly favourable and if specific provenances may be better adapted to the new conditions than others. Therefore, we estimated the potential early height growth under climate conditions in 2040–2060 for 20 beech provenances across a region covering the Czech Republic and Slovakia. This Central European region is expected to experience considerably drier and warmer conditions in the future. For this exercise, we implemented a new neural network model developed from height growth information obtained from the open-access BeechCOSTe52 database. The simulations are driven by past and future climate data obtained from the WorldClim database of historical climate data and future climate projections. Simulations revealed that provenances originating from drier regions performed on average significantly better than those from regions with good water supply. Moreover, provenances originating from drier regions had a particularly large advantage in the relatively arid regions of Central Czechia and Southern Slovakia. We can also confirm that all provenances showed a high phenotypic plasticity of height growth across the whole investigated region.

International audience; The vulnerability of forest species and tree populations to climate change is related to the exposure of the ecosystem to extreme climatic conditions and to the adaptive capacity of the population to cope with those conditions. Adaptive capacity is a relatively under-researched topic within the forest science community and there is an urgent need to understand to what extent particular combinations of traits have been shaped by natural selection under climatic gradients, potentially resulting in adaptive multi-trait associations. Thus, our aim was to quantify genetic variation in several leaf and woody traits that may contribute to multi-trait associations in which intraspecific variation could represent a source for species adaptation to climate change. A multi-trait approach was performed using nine Quercus petraea provenances originating from different locations that cover most of the species' distribution range over Europe and that were grown in a common garden. Multiple adaptive differences were observed between oak provenances but also some evolutionary stasis. Also, our results revealed higher genetic differentiation in traits related to phenology and growth than in those related to xylem anatomy, physiology and hydraulics for which no genetic differentiation was observed. The multiple associations between those traits and climate variables resulting from multivariate and path analyses suggest a multi-trait association largely involving phenological and growth traits for Quercus petraea. © The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Hydraulic failure has been extensively studied during drought-induced plant dieback, but its role in plant-pathogen interactions is under debate. During esca, a grapevine (Vitis vinifera) disease, symptomatic leaves are prone to irreversible hydraulic dysfunctions but little is known about the hydraulic integrity of perennial organs over the short- and long-term. We investigated the effects of esca on stem hydraulic integrity in naturally infected plants within a single season and across season(s). We coupled direct (ks) and indirect (kth) hydraulic conductivity measurements, and tylose and vascular pathogen detection with in vivo X-ray microtomography visualizations. Xylem occlusions (tyloses) and subsequent loss of stem hydraulic conductivity (ks) occurred in all shoots with severe symptoms (apoplexy) and in more than 60% of shoots with moderate symptoms (tiger-stripe), with no tyloses in asymptomatic shoots. In vivo stem observations demonstrated that tyloses occurred only when leaf symptoms appeared, and resulted in more than 50% loss of hydraulic conductance in 40% of symptomatic stems, unrelated to symptom age. The impact of esca on xylem integrity was only seasonal, with no long-term impact of disease history. Our study demonstrated how and to what extent a vascular disease such as esca, affecting xylem integrity, could amplify plant mortality through hydraulic failure.

In this work, the development of high luminescent europium(III) complexes in water solution is reported, including their synthesis, analyses of their photophysical properties and applications in bioassays. The three Eu(III) complexes are derived from new ligands based on a tripyridinophane platform. There are four distinct sections in the structure of these ligands: an 18-membered polyaminocarboxylic macrocycle to bind efficiently lanthanide ion in aqueous solutions, three chromophoric subunits (4-(phenylethynyl)pyridine moieties) to effectively sensitize the emission of the metal, two peripheral moieties to solubilise the complex in aqueous media (sulfonate, sulfobetaine or glucose groups) and a free NH2 group available for grafting or bioconjugation. In our synthetic procedure, a pivotal macrocyclic platform is obtained with a high yield in the crucial macrocyclization step due to a metal template ion effect (74% yield). In Tris aqueous buffer (pH 7.4), the Eu(III) complexes display a maximum excitation wavelength at 320 nm, a suitable overall quantum yield (14%), a relatively long lifetime (0.80 ms) and a one-photon brightness in the range of 10000 M-1 cm-1. Importantly, these photophysical properties are retained at dilute concentrations, even in the presence of a very large excess of potentially competing species, such as EDTA or Mg2+ ions. Furthermore, we report the bioconjugation of a Eu(III) complex labelled by an N-hydroxysuccinimide ester reactive group with an antibody (anti-Glutathione-S-Transferase) and the successful application of the corresponding antibody conjugate in the detection of GST-biotin in a fluoroimmunoassay. These new complexes provide solution for high sensitivity in Homogenous Time-Resolved Fluorescence (HTRF®) bioassays.

Chemokine receptors CXCR4 and ACKR3 are involved in multiple physiological and pathological processes. In overexpression systems, CXCR4 and ACKR3 form oligomers that are important for chemokine recognition and signaling. Whether this holds true in physiological settings is unknown as no evidence of such oligomers are available. To address this gap of knowledge, we developed a new approach for GPCR oligomer detection by time-resolved Forster resonance energy transfer (TR-FRET) between fluorescently labeled nanobodies. Fluorescent probes were conjugated to the C-terminus of extracellularly binding nanobodies using site-directed sortase-mediated labeling, retaining high affinity of the nanobodies for their target receptors. This approach, first validated in transfected cells by detecting the presence of CXCR4 and ACKR3 homomers and heteromers, reveals for the first time endogenous CXCR4 homomers in non-transfected human leukemia/lymphoma-derived cancer cell lines. More importantly, the investigations in peripheral blood mononuclear cells strongly support the existence of CXCR4 oligomers in native conditions.

Hydraulic failure has been extensively studied during drought-induced plant dieback, but its role in plant-pathogen interactions is under debate. During esca, a grapevine (Vitis vinifera) disease, symptomatic leaves are prone to irreversible hydraulic dysfunctions but little is known about the hydraulic integrity of perennial organs over the short- and long-term. We investigated the effects of esca on stem hydraulic integrity in naturally infected plants within a single season and across season(s). We coupled direct (ks) and indirect (kth) hydraulic conductivity measurements, and tylose and vascular pathogen detection with in vivo X-ray microtomography visualizations. We found xylem occlusions (tyloses), and subsequent loss of stemks, in all of the shoots with severe symptoms (apoplexy) and in more than 60% of the shoots with moderate symptoms (tiger-stripe), and no tyloses in shoots that were currently asymptomatic. In vivo stem observations demonstrated that tyloses were observed only when leaf symptoms appeared, and resulted in more than 50% PLC in 40% of symptomatic stems, unrelated to symptom age. The impact of esca on xylem integrity was only seasonal and no long-term impact of disease history was recorded. Our study demonstrated how and to what extent a vascular disease such as esca, affecting xylem integrity, could amplify plant mortality by hydraulic failure.HighlightOur study reveals that esca can critically affect xylem water movement in grapevine perennial organs, by the presence of plant-derived tyloses.

We report a detailed characterization of Eu3+ and Tb3+ complexes derived from a tripyridinophane macro cycle bearing three acetate side arms (H3tpptac). Tpptac3- displays an overall basicity (Σ log KiH) of 24.5, provides the formation of mononuclear ML species, and shows a good binding affinity for Ln3+ (log KLnL = 17.5-18.7). These complexes are also thermodynamically stable at physiological pH (pEu = 18.6, pTb = 18.0). It should be noted that the pGd value of Gd-tpptac (18.4) is only slightly lower than that of commercially available MRI contrast agents such as Gd-dota (pGd = 19.2). Moreover, a very good selectivity for these ions over the endogenous cations (log KCuL = 14.4, log KZnL = 12.9, and log KCaL = 9.3) is observed. The X-ray structure of the terbium complex shows the metal coordinated by the nine N6O3 donor set of the ligand and one inner-sphere water molecule. DFT calculations result in two Eu-tpptac structures with similar bond energies (ΔE = 0.145 eV): one structure in which the water is coordinated to the metal ion and one structure in which the water molecule is farther away from the ion, bound to the ligand with an OH-π bond. By detailed luminescence experiments, we demonstrate that the europium complex in aqueous solution presents a hydration equilibrium between nine-coordinate, dehydrated [Eu-tpptac]0 and ten-coordinate, monohydrated [Eu-tpptac(H2O)]0 species. A similar trend is observed for the terbium complex. Despite the presence of this hydration equilibrium, the H3tpptac ligand sensitizes Eu3+ and Tb3+ luminescence efficiently in buffered water at physiological pH. Particularly, the terbium complex displays a long excited-state lifetime of 2.24 ms and an overall quantum yield of 33% with a brightness of 3600 M-1 cm-1. Such features of Ln3+ complexes of H3tpptac indicate that this platform appears to be particularly appealing for the further development of luminescent lanthanide labels.

Xylem vulnerability to drought-induced embolism did not differ between stems and petioles of four woody species ( Betula pendula , Liriodendron tulipifera , Populus tremula and Olea europaea ). Our results, together with data compiled from published literature, indicate that hydraulic segmentation during drought stress is not consistently driven by difference in vulnerability to embolism between stem and terminal organs. Hydraulic failure and disconnection of distal organs during protracted drought stress is thought to protect large branches or trunks by reducing water loss and restricting the spread of embolism. Hydraulic segmentation and preferential sacrifice of distal organs such as leaves can be driven by two mechanisms: more negative water potentials at the terminal section of the hydraulic pathway and/or by higher vulnerability to xylem embolism of distal organs. Although vulnerability segmentation has been reported in the literature, the generality of this phenomenon is unclear, in part due to the methodological limitations related to direct measurement of xylem vulnerability to embolism in intact plants. The objective of this study was to evaluate vulnerability segmentation between petioles and stems using non-invasive micro computed tomography (microCT). Vulnerability to embolism was measured in leaf petioles and subtending stems of four woody species (Betula pendula R., Liriodendron tulipifera L., Populus tremula L. and Olea europaea L.) with contrasting drought tolerances. In addition, previously published vulnerability data for petioles and stems were compiled from the literature to investigate the commonality of hydraulic segmentation across a wide range of woody species, with the vulnerability curve methodology distinguished. Using non-invasive imaging on intact plants, we found no evidence of hydraulic segmentation between petioles and stems of four angiosperm tree species, regardless of mechanism. Moreover, the literature dataset indicated that little or no difference in vulnerability to embolism is present between petioles and stems when vulnerability curves were constructed using methods specifically measuring the dynamics of xylem tissue during dehydration (e.g. optical visualization, MicroCT). Our results suggest that vulnerability segmentation between stems and distal organs (petioles and leaves) is limited when only xylem tissue is considered. Large differences in vulnerability between stems and leaves are likely to be driven by extra-xylary components, rather than xylem embolism.

The majority of variation in six traits critical to the growth, survival and reproduction of plant species is thought to be organised along just two dimensions, corresponding to strategies of plant size and resource acquisition. However, it is unknown whether global plant trait relationships extend to climatic extremes, and if these interspecific relationships are confounded by trait variation within species. We test whether trait relationships extend to the cold extremes of life on Earth using the largest database of tundra plant traits yet compiled. We show that tundra plants demonstrate remarkably similar resource economic traits, but not size traits, compared to global distributions, and exhibit the same two dimensions of trait variation. Three quarters of trait variation occurs among species, mirroring global estimates of interspecific trait variation. Plant trait relationships are thus generalizable to the edge of global trait-space, informing prediction of plant community change in a warming world., It is unclear whether plant trait relationships found at the global scale extend to climatic extremes. Here the authors analyse six major aboveground traits to show that known plant trait relationships extend to the tundra biomes and exhibit the same two dimensions of variation detected at the global scale.

International audience; Climate change threatens food security, and plant science researchers have investigated methods of sustaining crop yield under drought. One approach has been to overproduce abscisic acid (ABA) to enhance water use efficiency. However, the concomitant effects of ABA overproduction on plant vascular system functioning are critical as it influences vulnerability to xylem hydraulic failure. We investigated these effects by comparing physiological and hydraulic responses to water deficit between a tomato (Solanum lycopersicum) wild type control (WT) and a transgenic line overproducing ABA (sp12). Under well-watered conditions, the sp12 line displayed similar growth rate and greater water use efficiency by operating at lower maximum stomatal conductance. X-ray microtomography revealed that sp12 was significantly more vulnerable to xylem embolism, resulting in a reduced hydraulic safety margin. We also observed a significant ontogenic effect on vulnerability to xylem embolism for both WT and sp12. This study demonstrates that the greater water use efficiency in the tomato ABA overproducing line is associated with higher vulnerability of the vascular system to embolism and a higher risk of hydraulic failure. Integrating hydraulic traits into breeding programmes represents a critical step for effectively managing a crop's ability to maintain hydraulic conductivity and productivity under water deficit.

Photosynthetic 'least-cost' theory posits that the optimal trait combination for a given environment is that where the summed costs of photosynthetic water and nutrient acquisition/use are minimised. The effects of soil water and nutrient availability on photosynthesis should be stronger as climate-related costs for both resources increase. Two independent datasets of photosynthetic traits, Globamax (1509 species, 288 sites) and Glob13C (3645 species, 594 sites), were used to quantify biophysical and biochemical limitations of photosynthesis and the key variable Ci /Ca (CO2 drawdown during photosynthesis). Climate and soil variables were associated with both datasets. The biochemical photosynthetic capacity was higher on alkaline soils. This effect was strongest at more arid sites, where water unit-costs are presumably higher. Higher values of soil silt and depth increased Ci /Ca , likely by providing greater H2 O supply, alleviating biophysical photosynthetic limitation when soil water is scarce. Climate is important in controlling the optimal balance of H2 O and N costs for photosynthesis, but soil properties change these costs, both directly and indirectly. In total, soil properties modify the climate-demand driven predictions of Ci /Ca by up to 30% at a global scale.

Drought represents a major abiotic constraint to plant growth and survival. On the one hand, plants keep stomata open for efficient carbon assimilation while, on the other hand, they close them to prevent permanent hydraulic impairment from xylem embolism. The order of occurrence of these two processes (stomatal closure and the onset of leaf embolism) during plant dehydration has remained controversial, largely due to methodological limitations. However, the newly developed optical visualization method now allows concurrent monitoring of stomatal behaviour and leaf embolism formation in intact plants. We used this new approach directly by dehydrating intact saplings of three contrasting tree species and indirectly by conducting a literature survey across a greater range of plant taxa. Our results indicate that increasing water stress generates the onset of leaf embolism consistently after stomatal closure, and that the lag time between these processes (i.e. the safety margin) rises with increasing embolism resistance. This suggests that during water stress, embolism-mediated declines in leaf hydraulic conductivity are unlikely to act as a signal for stomatal down-regulation. Instead, these species converge towards a strategy of closing stomata early to prevent water loss and delay catastrophic xylem dysfunction.

International audience; Cell surface receptors represent a vast majority of drug targets. Efforts have been conducted to develop biosensors reporting their conformational changes in live cells for pharmacological and functional studies. Although Förster resonance energy transfer (FRET) appears to be an ideal approach, its use is limited by the low signal-to-noise ratio. Here we report a toolbox composed of a combination of labeling technologies, specific fluorophores compatible with time-resolved FRET and a novel method to quantify signals. This approach enables the development of receptor biosensors with a large signal-to-noise ratio. We illustrate the usefulness of this toolbox through the development of biosensors for various G-protein-coupled receptors and receptor tyrosine kinases. These receptors include mGlu, GABAB, LH, PTH, EGF and insulin receptors among others. These biosensors can be used for high-throughput studies and also revealed new information on the activation process of these receptors in their cellular environment.

International audience; A goal of comparative physiology is to understand underlying causes of the tremendous diversity of land plant form and function. The hope is that functional diversity of land plants can be distilled to a few traits that together capture the essence of plant form and function (Díaz et al., 2016), thereby simplifying plant diversity into a tractable number of fundamental ecophysiological ‘strategies’ (or plant functional types). However, there has been disagreement and uncertainty as to (1) which traits should make the shortlist – that is what the key dimensions of trait covariation are, and (2) whether broad‐scale trait patterns and the inferred functional tradeoffs hold at smaller scales relevant to predicting species responses to global change (e.g. whether between species trait patterns hold within individual species). In this issue of New Phytologist, the article by Rosas et al. (pp. 632–646) is a thought‐provoking article that investigates within‐ and among‐species variation in key plant traits along a water availability gradient in two major plant families from a European forest.

Vascular pathogens cause disease in a large spectrum of perennial plants, with leaf scorch being one of the most conspicuous symptoms. Esca in grapevine (Vitis vinifera) is a vascular disease with huge negative effects on grape yield and the wine industry. One prominent hypothesis suggests that vascular disease leaf scorch is caused by fungal pathogen-derived elicitors and toxins. Another hypothesis suggests that leaf scorch is caused by hydraulic failure due to air embolism, the pathogen itself, and/or plant-derived tyloses and gels. In this study, we transplanted mature, naturally infected esca symptomatic vines from the field into pots, allowing us to explore xylem integrity in leaves (i.e. leaf midveins and petioles) using synchrotron-based in vivo x-ray microcomputed tomography and light microscopy. Our results demonstrated that symptomatic leaves are not associated with air embolism. In contrast, symptomatic leaves presented significantly more nonfunctional vessels resulting from the presence of nongaseous embolisms (i.e. tyloses and gels) than control leaves, but there was no significant correlation with disease severity. Using quantitative PCR, we determined that two vascular pathogen species associated with esca necrosis in the trunk were not found in leaves where occlusions were observed. Together, these results demonstrate that symptom development is associated with the disruption of vessel integrity and suggest that symptoms are elicited at a distance from the trunk where fungal infections occur. These findings open new perspectives on esca symptom expression where the hydraulic failure and elicitor/toxin hypotheses are not necessarily mutually exclusive.

Background and aimsHydraulic studies are currently biased towards conifers and dicotyledonous angiosperms; responses of arborescent monocots to increasing temperature and drought remain poorly known. This study aims to assess xylem resistance to drought-induced embolism in palms.MethodsWe quantified embolism resistance via P50 (xylem pressure inducing 50 % embolism or loss of hydraulic conductivity) in petioles and leaflets of six palm species differing in habitat and phylogenetic relatedness using three techniques: in vivo X-ray-based microcomputed tomography, the in situ flow centrifuge technique and the optical vulnerability method.Key resultsOur results show that P50 of petioles varies greatly in the palm family, from −2.2 ± 0.4 MPa in Dypsis baronii to −5.8 ± 0.3 MPa in Rhapis excelsa (mean ± s.e.). No difference or weak differences were found between petioles and leaf blades within species. Surprisingly, where differences occurred, leaflets were less vulnerable to embolism than petioles. Embolism resistance was not correlated with conduit size (r = 0.37, P = 0.11).ConclusionsThis study represents the first estimate of drought-induced xylem embolism in palms across biomes and provides the first step towards understanding hydraulic adaptations in long-lived arborescent monocots. It showed an almost 3-fold range of embolism resistance between palm species, as large as that reported in all angiosperms. We found little evidence for hydraulic segmentation between leaflets and petioles in palms, suggesting that when it happens, hydraulic segregation may lack a clear relationship with organ cost or replaceability.

A relatively high resistance of leaf xylem to embolism may not explain hydraulic segmentation between leaves and branches in angiosperms., According to the hydraulic vulnerability segmentation hypothesis, leaves are more vulnerable to decline of hydraulic conductivity than branches, but whether stem xylem is more embolism resistant than leaves remains unclear. Drought-induced embolism resistance of leaf xylem was investigated based on X-ray microcomputed tomography (microCT) for Betula pendula, Laurus nobilis, and Liriodendron tulipifera, excluding outside-xylem, and compared with hydraulic vulnerability curves for branch xylem. Moreover, bordered pit characters related to embolism resistance were investigated for both organs. Theoretical P50 values (i.e. the xylem pressure corresponding to 50% loss of hydraulic conductance) of leaves were generally within the same range as hydraulic P50 values of branches. P50 values of leaves were similar to branches for L. tulipifera (−2.01 versus −2.10 MPa, respectively), more negative for B. pendula (−2.87 versus −1.80 MPa), and less negative for L. nobilis (−6.4 versus −9.2 MPa). Despite more narrow conduits in leaves than branches, mean interconduit pit membrane thickness was similar in both organs, but significantly higher in leaves of B. pendula than in branches. This case study indicates that xylem shows a largely similar embolism resistance across leaves and branches, although differences both within and across organs may occur, suggesting interspecific variation with regard to the hydraulic vulnerability segmentation hypothesis.

A series of four new 18-membered hexaaza macrocyclic ligands bearing three endocyclic pyridine units and acetate or methylenephosphonate pendant arms has been prepared. The new synthetic procedure is based on the use of amine and diamine precursors incorporating masked carboxylate or phosphonate functions and on an efficient sodium template effect which controls the crucial macrocyclization step (yields of macrocyclization reactions: 62–88%). This procedure appears as a suitable alternative compared to the classical Richman-Atkins methodology generally used for the preparation of this class of macrocycles. As demonstrated with the EuIII and TbIII complexes derived from two ligands, these tripyridinophane chelators form luminescent and stable mononuclear LnIII complexes in aqueous solution at physiological pH. In such a medium, TbIII complexes exhibit a brightness of 1700 (λexc = 279 nm) and 3000 (λexc = 268 nm) M−1 cm−1.

International audience; Methods to estimate xylem embolism resistance generally rely on hydraulic measurements, which can be far from straightforward. Recently, a pneumatic method based on air flow measurements of terminal branch ends was proposed to construct vulnerability curves by linking the amount of air extracted from a branch with the degree of embolism. We applied this novel technique for 10 temperate tree species, including six diffuse, two ring-porous and two gymnosperm species, and compared the pneumatic curves with hydraulic ones obtained from either the flow-centrifuge or the hydraulic-bench dehydration method. We found that the pneumatic method provides a good estimate of the degree of xylem embolism for all angiosperm species. The xylem pressure at 50% and 88% loss of hydraulic conductivity (i.e., Ψ50 and Ψ88) based on the methods applied showed a strongly significant correlation for all eight angiosperms. However, the pneumatic method showed significantly reduced Ψ50 values for the two conifers. Our findings suggest that the pneumatic method could provide a fast and accurate approach for angiosperms due to its convenience and feasibility, at least within the range of embolism resistances covered by our samples.

Altres ajuts europeus: P.A.W. was additionally supported by the European Union Fourth Environment and Climate Framework Programme (Project Number ENV4-CT970586)P.A.W. was additionally supported by the European Union Fourth Environment and Climate Framework Programme (Project Number ENV4-CT970586). The tundra is warming more rapidly than any other biome on Earth, and the potential ramifications are far-reaching because of global feedback effects between vegetation and climate. A better understanding of how environmental factors shape plant structure and function is crucial for predicting the consequences of environmental change for ecosystem functioning. Here we explore the biome-wide relationships between temperature, moisture and seven key plant functional traits both across space and over three decades of warming at 117 tundra locations. Spatial temperature-trait relationships were generally strong but soil moisture had a marked influence on the strength and direction of these relationships, highlighting the potentially important influence of changes in water availability on future trait shifts in tundra plant communities. Community height increased with warming across all sites over the past three decades, but other traits lagged far behind predicted rates of change. Our findings highlight the challenge of using space-for-time substitution to predict the functional consequences of future warming and suggest that functions that are tied closely to plant height will experience the most rapid change. They also reveal the strength with which environmental factors shape biotic communities at the coldest extremes of the planet and will help to improve projections of functional changes in tundra ecosystems with climate warming.

Key message: Direct, non-invasive X-ray microtomography and optical technique observations applied in stems and leaves of intact seedlings revealed that laurel is highly resistant to drought-induced xylem embolism. Contrary to what has been brought forward, daily cycles of embolism formation and refilling are unlikely to occur in this species and to explain how it copes with drought. Context: There has been considerable controversy regarding xylem embolism resistance for long-vesselled angiosperm species and particularly for the model species for refilling (Laurus nobilis L.). Aims: The purpose of this study was to resolve the hydraulic properties of this species by documenting vulnerability curves of different organs in intact plants. Methods: Here, we applied a direct, non-invasive method to visualize xylem embolism in stems and leaves of intact laurel seedlings up to 2-m tall using X-ray microtomography (microCT) observations and the optical vulnerability technique. These approaches were coupled with complementary centrifugation measurements performed on 1-m long branches sampled from adult trees and compared with additional microCT analyses carried out on 80-cm cut branches. Results: Direct observations of embolism spread during desiccation of intact laurels revealed that 50% loss of xylem conductivity (Psi(50)) was reached at - 7.9 +/- 0.5 and - 8.4 +/- 0.3 MPa in sterns and leaves, respectively, while the minimum xylem water potentials measured in the field were - 4.2 MPa during a moderate drought season. Those findings reveal that embolism formation is not routine in Laurus nobilis contrary to what has been previously reported. These Psi(50) values were close to those based on the flow-centrifuge technique (- 9.2 +/- 0.2 MPa), but at odds with microCT observations of cut branches (- 4.0 +/- 0.5 MPa). Conclusion: In summary, independent methods converge toward the same conclusion that laurel is highly resistant to xylem embolism regardless its development stage. Under typical growth conditions without extreme drought events, this species maintains positive hydraulic safety margin, while daily cycles of embolism formation and refilling are unlikely to occur in this species.

Drought-induced xylem embolism is a key process closely related to plantmortality during extreme drought events. However, this process has beenpoorly investigated in crop species to date, despite the observed declineof crop productivity under extreme drought conditions. Interspecific variation in hydraulic traits has frequently been reported, but less is knownabout intraspecific variation in crops. We assessed the intraspecific variability of embolism resistance in four sunflower (Helianthus annuus L.) accessions grown in well-watered conditions. Vulnerability to embolism was deter-mined by the in situ flow-centrifuge method (cavitron), and possible trade-offsbetween xylem safety, xylem efficiency and growth were assessed. The relationship between stem anatomy and hydraulic traits was also investigated.Mean P50 was −3 MPa, but significant variation was observed between accessions, with values ranging between −2.67 and −3.22 MPa. Embolism resistance was negatively related to growth and positively related to xylem-specifichydraulic conductivity. There is, therefore, a trade-off between hydraulic safetyand growth but not between hydraulic safety and efficiency. Finally, we foundthat a few anatomical traits, such as vessel density and the area of the vessellumen relative to that of the secondary xylem, were related to embolism resistance, whereas stem tissue lignification was not. Further investigations are nowrequired to investigate the link between the observed variability of embolismresistance and yield, to facilitate the identification of breeding strategies toimprove yields in an increasingly arid world.

International audience; The main inhibitory neurotransmitter, γ-aminobutyric acid (GABA), modulates many synapses by activating the G protein-coupled receptor GABAB, which is a target for various therapeutic applications. It is an obligatory heterodimer made of GB1 and GB2 that can be regulated by positive allosteric modulators (PAMs). The molecular mechanism of activation of the GABAB receptor remains poorly understood. Here, we have developed FRET-based conformational GABAB sensors compatible with high-throughput screening. We identified conformational changes occurring within the extracellular and transmembrane domains upon receptor activation, which are smaller than those observed in the related metabotropic glutamate receptors. These sensors also allow discrimination between agonists of different efficacies and between PAMs that have different modes of action, which has not always been possible using conventional functional assays. Our study brings important new information on the activation mechanism of the GABAB receptor and should facilitate the screening and identification of new chemicals targeting this receptor.

Nurse-plants generally have positive effects on understorey species by creating more suitable conditions for stress-intolerant plants relative to open micro-habitats. However, long-term effects of this plant-plant facilitation system have been rarely examined. Seeds of five desert annual species from Atiquipa coastal desert in Southern Peru were used to examine whether different microenvironmental conditions under the nurse-plants Caesalpinia spinosa Molina (Kuntze) lead to differences in seed biology and germinability of annual plants relative to open, canopy-free conditions. Seeds collected from plants associated with nurse-plants were predicted to be (i) larger due to more favourable growing conditions, (ii) more viable and with greater germination rates, (iii) less variable in size and viability due to reduced environmental heterogeneity, and (iv) to germinate faster to avoid apparent competition with other annuals. Seed attribute measurements and germination trials in growth chambers were used to test these predictions. Although the plant abundance of only 2 of 5 species was strongly facilitated by the nurse-plant, no significant differences were found in seed mass, viability or relative variability between understorey and open micro-habitats for any of the species. Contrary to our predictions, final seed germination rates of seeds from open micro-habitats were higher, and the open micro-habitat treatment was more favourable for germination of seeds from both open and understorey environments. Taken together, these results suggest that plant-plant facilitation does not necessarily affect seed biology traits. Further studies addressing larger distribution ranges and/or density gradients of understorey species will illuminate the potential evolutionary effects of nurse-plants.

International audience; Trees act as ecosystem engineers and invasions by exotic tree species profoundly impact recipient communities. Recently, research on invasive trees has dramatically increased, enabling the assessment of general trends in tree invasion. Analysing 90 studies dealing with 45 invasive tree species, we conducted a quantitative review and a meta-analysis to estimate the relevance of eight leading hypotheses for explaining tree invasions. We also tested whether species functional traits (growth rate, density/cover, germination, biomass and survival) equally promote tree invasiveness. Overall, our results suggest that several hypotheses, linked to invasibility or invasiveness, are pertinent to explain tree invasions. Furthermore, more than one hypothesis has been supported for a given species, which indicates that multiple factors lead to the success of invasive tree species. In addition, growth rate appears to be the most efficient predictor of invasiveness for invasive trees and could thus be used as a means to identify potential alien tree invasions. We conclude that further investigations are needed to test the consistency of some hypotheses across a broader pool of invasive tree species, whilst experimental studies with the same tree species across a larger range of sites would help to reveal the full suite of factors that affect tree invasions.

Seven-transmembrane domain (7TM) receptors have important functions in cell–cell communication and can assemble into dimers or oligomers. Such complexes may allow specific functional cross-talk through trans-activation of interacting 7TMs, but this hypothesis requires further validation. Herein, we used the GABAB receptor, which is composed of two distinct subunits, GABAB1, which binds the agonist, and GABAB2, which activates G proteins, as a model system. By using a novel orthogonal-labelling approach compatible with time-resolved FRET and based on ACP- and SNAP-tag technologies to verify the heterodimerization of wild-type and mutated GABAB subunits, we demonstrate the existence of a direct allosteric coupling between the 7TMs of GABAB heterodimers. Indeed, a GABAB receptor, in which the GABAB2 extracellular domain was deleted, was still capable of activating G proteins. Furthermore, synthetic ligands for the GABAB2 7TM could increase agonist affinity at the GABAB1 subunit in this mutated receptor. In addition to bringing new information on GABAB receptor activation, these data clearly demonstrate the existence of direct trans-activation between the 7TM of two interacting proteins.

Acer negundo (boxelder maple) is a North American native tree species that currently invades riparian and disturbed areas in Europe, affecting both bank stability and ecosystem biodiversity. As a response to managers’ requests, we aimed at finding an eco-friendly method which would locally remove this species and help habitat restoration. Four control methods were tested on A . negundo adults and saplings from stands located in three experimental sites along different watercourses in Southwestern France: girdling, low cutting, high cutting, and cutting followed by the application of juglone (a natural allelopathic substance from walnut tree leaves). Mortality and resprout production on the treated A. negundo individuals were assessed during two years following the application of the control methods. Girdling was the most efficient method as it significantly induced higher mortality rates compared to the others (65 vs 15% of dead A . negundo two years after treatment administration). When healing emerged on trunks, yearly repeated girdling was required to reach full success. None of the control methods significantly reduced resprout production; not even the application of juglone. Girdling is the most recommended method to kill and remove A. negundo at a local scale in invaded natural habitats. Considering that A. negundo benefits from increases in light availability to outcompete native species, we further recommend removing seedlings from understories when applying girdling on adult and sapling individuals in order to optimize restoration conditions in natural stands and improve native species re-establishment.

The development of the brightest luminescent europium(III) complexes is traced, including analysis of the C3-symmetric core complex based on a functionalized triazacyclononane and identification of the most suitable strongly absorbing chromophore. Strategies for the synthesis of the complexes, including enantiopure analogues, are outlined and opportunities for applications in time-resolved microscopy and spectral imaging emphasised. Practicable examples are introduced, including selective organelle staining for cellular optical imaging at 65 nm resolution and the development of new bioassays using time-resolved FRET methods.

International audience; Genetically based phenotypic differentiation between native and invasive populations of exotic plants has been increasingly documented and commonly invoked to explain the success of some invasive species. Nonetheless, this basic information is lacking for invasive trees although they currently represent a major concern worldwide. Reciprocal common gardens were therefore set up in both native and introduced ranges of two exotic maple trees to assess the contribution of genetic differentiation and phenotypic plasticity to tree invasiveness. Almost 3,000 native and invasive seedlings of Acer negundo and Acer platanoides were planted in Canada and in France and their performances were compared in various life-history traits related to growth, leaf phenology and ecophysiology over 2 and 3 year periods. Invasive populations of A. negundo exhibited strong genetic differentiation in all the traits examined. Compared to their native conspecifics, they grew significantly larger in the introduced range and showed lower survival, reduced maximum assimilation rate and increased leaf area in the two gardens. They also expressed greater plasticity for growth and greater phenological sensitivity to temperature. Native and invasive populations of A. platanoides were plastic across environments but in contrast did not exhibit any genetic differentiation. This cross-continental comparison provides evidence that both genetic differentiation and phenotypic plasticity contribute synergistically to tree invasiveness. The influence of these respective processes depends on stage of invasion and the life-history strategy of each species. Plastic effects are likely more important during colonization and establishment whilst genetic effects may contribute more significantly during the spread of established populations.

The development of the brightest luminescent europium(III) complexes is traced, including analysis of the C3-symmetric core complex based on a functionalized triazacyclononane and identification of the most suitable strongly absorbing chromophore. Strategies for the synthesis of the complexes, including enantiopure analogues, are outlined and opportunities for applications in time-resolved microscopy and spectral imaging emphasised. Practicable examples are introduced, including selective organelle staining for cellular optical imaging at 65 nm resolution and the development of new bioassays using time-resolved FRET methods.

A series of europium and terbium complexes based on a functionalized triazacyclononane carboxylate or phosphinate macrocyclic ligand is described. The influence of the anionic group, that is, carboxylate, methylphosphinate, or phenylphosphinate, on the photophysical properties was studied and rationalized on the basis of DFT calculated structures. The nature, number, and position of electron-donating or electron-withdrawing aryl substituents were varied systematically within the same phenylethynyl scaffold in order to optimize the brightness of the corresponding europium complexes and investigate their two-photon absorption properties. Finally, the europium complexes were examined in cell-imaging applications, and selected terbium complexes were studied as potential oxygen sensors.

Luminescent europium complexes are used in a broad range of applications as a result of their particular emissive properties. The synthesis and application of bright, highly water-soluble, and negatively charged sulfonic- or carboxylic acid derivatives of para-substituted aryl–alkynyl triazacyclononane complexes are described. Introduction of the charged solubilizing moieties suppresses cellular uptake or adsorption to living cells making them applicable for labeling and performing assays on membrane receptors. These europium complexes are applied to monitor fluorescent ligand binding on cell-surface proteins with time-resolved Forster resonance energy transfer (TR-FRET) assays in plate-based format and using TR-FRET microscopy.

International audience; Although the genetic aspects of biological invasions are receiving more attention in the scientific literature, analyses of phenotypic plasticity and genotype-by-environment interactions are still seldom considered in tree invasion biology. Previous studies have shown that invasions of tree species can be affected by intraspecific phenotypic plasticity, pre-adaptation, and post-introduction evolution, and we suggest there are opportunities for new developments in this field. Here, we present a description of the use of quantitative and molecular genetics in tree invasion biology, and propose an approach based on common garden experiments, quantitative and molecular genetic methods to investigate the role of adaptive evolution, phenotypic plasticity, and genotype-by-environment interactions in tree invasions, particularly at the infraspecific level. We illustrate the utility of this approach using examples from quantitative genetic studies of Pinus and an example from a classical reciprocal common garden experiment with Acer species. By using this approach, researchers can test hypotheses about the role and strength of genetic and environmental effects on population-level invasiveness and gain insights into evolutionary processes that occur during biological invasions. Moreover, knowledge of phenotypic plasticity and local adaption of tree populations may help researchers improve assessments of invasion risk.

To maintain homeostasis, hypothalamic neurons in the arcuate nucleus must dynamically sense and integrate a multitude of peripheral signals. Blood-borne molecules must therefore be able to circumvent the tightly sealed vasculature of the blood–brain barrier to rapidly access their target neurons. However, how information encoded by circulating appetite-modifying hormones is conveyed to central hypothalamic neurons remains largely unexplored. Using in vivo multiphoton microscopy together with fluorescently labeled ligands, we demonstrate that circulating ghrelin, a versatile regulator of energy expenditure and feeding behavior, rapidly binds neurons in the vicinity of fenestrated capillaries, and that the number of labeled cell bodies varies with feeding status. Thus, by virtue of its vascular connections, the hypothalamus is able to directly sense peripheral signals, modifying energy status accordingly.

International audience; Phenotypic plasticity is a key mechanism associated with the spread of exotic plants and previous studies have found that invasive species are generally more plastic than co-occurring species. Comparatively, the evolution of phenotypic plasticity in plant invasion has received less attention, and in particular, the genetic basis of plasticity is largely unexamined. Native from North America, Acer negundo L. is aggressively impacting the riparian forests of southern and eastern Europe thanks to higher plasticity relative to co-occurring native species. We therefore tested here whether invasive populations have evolved increased plasticity since introduction. The performance of 1152 seedlings from 8 native and 8 invasive populations was compared in response to nutrient availability. Irrespective of nutrients, invasive populations had higher growth and greater allocation to above-ground biomass relative to their native conspecifics. More importantly, invasive genotypes did not show increased plasticity in any of the 20 traits examined. This result suggests that the high magnitude of plasticity to nutrient variation of invasive seedlings might be pre-adapted in the native range. Invasiveness of A. negundo could be explained by higher mean values of traits due to genetic differentiation rather than by evolution of increased plasticity.