No evidence of canopy-scale leaf thermoregulation to cool leaves below air temperature across a range of forest ecosystems.

Understanding and predicting the relationship between leaf temperature (Tleaf) and air temperature (Tair) is essential for projecting responses to a warming climate, as studies suggest that many forests are near thermal thresholds for carbon uptake. Based on leaf measurements, the limited leaf homeothermy hypothesis argues that daytime Tleaf is maintained near photosynthetic temperature optima and below damaging temperature thresholds. Specifically, leaves should cool below Tair at higher temperatures (i.e., > ∼25-30°C) leading to slopes <1 in Tleaf/Tair relationships and substantial carbon uptake when leaves are cooler than air. This hypothesis implies that climate warming will be mitigated by a compensatory leaf cooling response. A key uncertainty is understanding whether such thermoregulatory behavior occurs in natural forest canopies. We present an unprecedented set of growing season canopy-level leaf temperature (Tcan) data measured with thermal imaging at multiple well-instrumented forest sites in North and Central America. Our data do not support the limited homeothermy hypothesis: canopy leaves are warmer than air during most of the day and only cool below air in mid to late afternoon, leading to Tcan/Tair slopes >1 and hysteretic behavior. We find that the majority of ecosystem photosynthesis occurs when canopy leaves are warmer than air. Using energy balance and physiological modeling, we show that key leaf traits influence leaf-air coupling and ultimately the Tcan/Tair relationship. Canopy structure also plays an important role in Tcan dynamics. Future climate warming is likely to lead to even greater Tcan, with attendant impacts on forest carbon cycling and mortality risk.

Natural variation identifies new effectors of water-use efficiency in Arabidopsis.

Water-use efficiency (WUE) is the ratio of biomass produced per unit of water consumed; thus, it can be altered by genetic factors that affect either side of the ratio. In the present study, we exploited natural variation for WUE to discover loci affecting either biomass accumulation or water use as factors affecting WUE. Genome-wide association studies (GWAS) using integrated WUE measured through carbon isotope discrimination (δ13C) of Arabidopsis thaliana accessions identified genomic regions associated with WUE. Reverse genetic analysis of 70 candidate genes selected based on the GWAS results and transcriptome data identified 25 genes affecting WUE as measured by gravimetric and δ13C analyses. Mutants of four genes had higher WUE than wild type, while mutants of the other 21 genes had lower WUE. The differences in WUE were caused by either altered biomass or water consumption (or both). Stomatal density (SD) was not a primary cause of altered WUE in these mutants. Leaf surface temperatures indicated that transpiration differed for mutants of 16 genes, but generally biomass accumulation had a greater effect on WUE. The genes we identified are involved in diverse cellular processes, including hormone and calcium signaling, meristematic activity, photosynthesis, flowering time, leaf/vasculature development, and cell wall composition; however, none of them had been previously linked to WUE. Thus, our study successfully identified effectors of WUE that can be used to understand the genetic basis of WUE and improve crop productivity.

Fast Assimilation-Temperature Response: a FAsTeR method for measuring the temperature dependence of leaf-level photosynthesis.

We present the Fast Assimilation-Temperature Response (FAsTeR) method, a new method for measuring plant assimilation-temperature (AT) response that reduces measurement time and increases data density compared with conventional methods. The FAsTeR method subjects plant leaves to a linearly increasing temperature ramp while taking rapid, nonequilibrium measurements of gas exchange variables. Two postprocessing steps are employed to correct measured assimilation rates for nonequilibrium effects and sensor calibration drift. Results obtained with the new method are compared with those from two conventional stepwise methods. Our new method accurately reproduces results obtained from conventional methods, reduces measurement time by a factor of c. 3.3 (from c. 90 to 27 min), and increases data density by a factor of c. 55 (from c. 10 to c. 550 observations). Simulation results demonstrate that increased data density substantially improves confidence in parameter estimates and drastically reduces the influence of noise. By improving measurement speed and data density, the FAsTeR method enables users to ask fundamentally new kinds of ecological and physiological questions, expediting data collection in short-field campaigns, and improving the representativeness of data across species in the literature.

Impacts of elevated temperature and vapour pressure deficit on leaf gas exchange and plant growth across six tropical rainforest tree species.

Elevated air temperature (Tair) and vapour pressure deficit (VPDair) significantly influence plant functioning, yet their relative impacts are difficult to disentangle. We examined the effects of elevated Tair (+6°C) and VPDair (+0.7 kPa) on the growth and physiology of six tropical tree species. Saplings were grown under well-watered conditions in climate-controlled glasshouses for 6 months under three treatments: (1) low Tair and low VPDair, (2) high Tair and low VPDair, and (3) high Tair and high VPDair. To assess acclimation, physiological parameters were measured at a set temperature. Warm-grown plants grown under elevated VPDair had significantly reduced stomatal conductance and increased instantaneous water use efficiency compared to plants grown under low VPDair. Photosynthetic biochemistry and thermal tolerance (Tcrit) were unaffected by VPDair, but elevated Tair caused Jmax25 to decrease and Tcrit to increase. Sapling biomass accumulation for all species responded positively to an increase in Tair, but elevated VPDair limited growth. This study shows that stomatal limitation caused by even moderate increases in VPDair can decrease productivity and growth rates in tropical species independently from Tair and has important implications for modelling the impacts of climate change on tropical forests.

Canopy temperatures strongly overestimate leaf thermal safety margins of tropical trees.

Current estimates of temperature effects on plants mostly rely on air temperature, although it can significantly deviate from leaf temperature (Tleaf). To address this, some studies have used canopy temperature (Tcan). However, Tcan fails to capture the fine-scale variation in Tleaf among leaves and species in diverse canopies. We used infrared radiometers to study Tleaf and Tcan and how they deviate from air temperature (ΔTleaf and ΔTcan) in multispecies tropical tree plantations at three sites along an elevation and temperature gradient in Rwanda. Our results showed high Tleaf (up to c. 50°C) and ΔTleaf (on average 8-10°C and up to c. 20°C) of sun-exposed leaves during 10:00 h-15:00 h, being close to or exceeding photosynthetic heat tolerance thresholds. These values greatly exceeded simultaneously measured values of Tcan and ΔTcan, respectively, leading to strongly overestimated leaf thermal safety margins if basing those on Tcan data. Stomatal conductance and leaf size affected Tleaf and Tcan in line with their expected influences on leaf energy balance. Our findings highlight the importance of leaf traits for leaf thermoregulation and show that monitoring Tcan is not enough to capture the peak temperatures and heat stress experienced by individual leaves of different species in tropical forest canopies.

Caught between two states: The compromise in acclimation of photosynthesis, transpiration and mesophyll conductance to different amplitudes of fluctuating irradiance.

While dynamic regulation of photosynthesis in fluctuating light is increasingly recognized as an important driver of carbon uptake, acclimation to realistic irradiance fluctuations is still largely unexplored. We subjected Arabidopsis thaliana (L.) wild-type and jac1 mutants to irradiance fluctuations with distinct amplitudes and average irradiance. We examined how irradiance fluctuations affected leaf structure, pigments and physiology. A wider amplitude of fluctuations produced a stronger acclimation response. Large reductions of leaf mass per area under fluctuating irradiance framed our interpretation of changes in photosynthetic capacity and mesophyll conductance as measured by three separate methods, in that photosynthetic investment increased markedly on a mass basis, but only a little on an area basis. Moreover, thermal imagery showed that leaf transpiration was four times higher under fluctuating irradiance. Leaves growing under fluctuating irradiance, although thinner, maintained their photosynthetic capacity, as measured through light- and CO2-response curves; suggesting their photosynthesis may be more cost-efficient than those under steady light, but overall may incur increased maintenance costs. This is especially relevant for plant performance globally because naturally fluctuating irradiance creates conflicting acclimation cues for photosynthesis and transpiration that may hinder progress towards ensuring food security under climate-related extremes of water stress.

Ecotypic Variation in Leaf Thermoregulation and Heat Tolerance but Not Thermal Safety Margins in Tropical Trees.

To avoid reaching lethal temperatures during periods of heat stress, plants may acclimate either their biochemical thermal tolerance or leaf morphological and physiological characteristics to reduce leaf temperature (Tleaf). While plants from warmer environments may have a greater capacity to regulate Tleaf, the extent of intraspecific variation and contribution of provenance is relatively unexplored. We tested whether upland and lowland provenances of four tropical tree species grown in a common garden differed in their thermal safety margins by measuring leaf thermal traits, midday leaf-to-air temperature differences (∆Tleaf) and critical leaf temperatures defined by chlorophyll fluorescence (Tcrit). Provenance variation was species- and trait-specific. Higher ∆Tleaf and Tcrit were observed in the lowland provenance for Terminalia microcarpa, and in the upland provenance for Castanospermum australe, with no provenance effects in the other two species. Within-species covariation of Tcrit and ∆Tleaf led to a convergence of thermal safety margins across provenances. While future studies should expand the number of provenances and species investigated, our findings suggest that lowland and upland provenances may not differ substantially in their vulnerability to heat stress, as determined by thermal safety margins, despite differences in operating temperatures and Tcrit.

High heat tolerance, evaporative cooling, and stomatal decoupling regulate canopy temperature and their safety margins in three European oak species.

Heatwaves and soil droughts are increasing in frequency and intensity, leading many tree species to exceed their thermal thresholds, and driving wide-scale forest mortality. Therefore, investigating heat tolerance and canopy temperature regulation mechanisms is essential to understanding and predicting tree vulnerability to hot droughts. We measured the diurnal and seasonal variation in leaf water potential (Ψ), gas exchange (photosynthesis Anet and stomatal conductance gs), canopy temperature (Tcan), and heat tolerance (leaf critical temperature Tcrit and thermal safety margins TSM, i.e., the difference between maximum Tcan and Tcrit) in three oak species in forests along a latitudinal gradient (Quercus petraea in Switzerland, Quercus ilex in France, and Quercus coccifera in Spain) throughout the growing season. Gas exchange and Ψ of all species were strongly reduced by increased air temperature (Tair) and soil drying, resulting in stomatal closure and inhibition of photosynthesis in Q. ilex and Q. coccifera when Tair surpassed 30°C and soil moisture dropped below 14%. Across all seasons, Tcan was mainly above Tair but increased strongly (up to 10°C > Tair) when Anet was null or negative. Although trees endured extreme Tair (up to 42°C), positive TSM were maintained during the growing season due to high Tcrit in all species (average Tcrit of 54.7°C) and possibly stomatal decoupling (i.e., Anet ≤0 while gs >0). Indeed, Q. ilex and Q. coccifera trees maintained low but positive gs (despite null Anet), decreasing Ψ passed embolism thresholds. This may have prevented Tcan from rising above Tcrit during extreme heat. Overall, our work highlighted that the mechanisms behind heat tolerance and leaf temperature regulation in oak trees include a combination of high evaporative cooling, large heat tolerance limits, and stomatal decoupling. These processes must be considered to accurately predict plant damages, survival, and mortality during extreme heatwaves.

Climate Constrains Photosynthetic Strategies in Darwin's Daisies: A Test of the Climatic Variability and Jack-of-All-Trades Hypotheses.

AbstractEmpirical evidence for the climate variability and performance trade-off hypotheses is limited to animals, and it is unclear whether climate constrains the photosynthetic strategies of plants. The plant genus Scalesia Arn. ex Lindl (family Asteraceae), endemic to the Galápagos archipelago, provides an ideal study system to test these hypotheses because of its species with markedly different leaf morphologies that occupy distinct climatic zones. In this study we tested the classic hypotheses that (1) climate constrains leaf size, (2) high climatic temperature variability selects for thermal generalists (i.e., the climate variability hypothesis), and (3) there is a trade-off between the breadth and rate of photosynthetic performance (i.e., jack-of-all-trades but master of none hypothesis). To do this we measured the leaf morphologies and photosynthetic temperature response curves of 11 Scalesia species. In support of a priori predictions, we found that small-leaved Scalesia species were more likely to occupy hotter and drier climates than large-leaved species, there was a positive relationship between climatic temperature variability and the breadth of photosynthetic performance, and photosynthetic performance was negatively correlated with photosynthetic breadth. Our study is among the first to provide evidence for the performance-breadth trade-off hypothesis in photosynthesis, suggesting that climate change may select for photosynthetic thermal generalists.

Thermal sensitivity across forest vertical profiles: patterns, mechanisms, and ecological implications.

Rising temperatures are influencing forests on many scales, with potentially strong variation vertically across forest strata. Using published research and new analyses, we evaluate how microclimate and leaf temperatures, traits, and gas exchange vary vertically in forests, shaping tree, and ecosystem ecology. In closed-canopy forests, upper canopy leaves are exposed to the highest solar radiation and evaporative demand, which can elevate leaf temperature (Tleaf ), particularly when transpirational cooling is curtailed by limited stomatal conductance. However, foliar traits also vary across height or light gradients, partially mitigating and protecting against the elevation of upper canopy Tleaf . Leaf metabolism generally increases with height across the vertical gradient, yet differences in thermal sensitivity across the gradient appear modest. Scaling from leaves to trees, canopy trees have higher absolute metabolic capacity and growth, yet are more vulnerable to drought and damaging Tleaf than their smaller counterparts, particularly under climate change. By contrast, understory trees experience fewer extreme high Tleaf 's but have fewer cooling mechanisms and thus may be strongly impacted by warming under some conditions, particularly when exposed to a harsher microenvironment through canopy disturbance. As the climate changes, integrating the patterns and mechanisms reviewed here into models will be critical to forecasting forest-climate feedback.

Limits of thermal and hydrological tolerance in a foundation tree species (Populus fremontii) in the desert southwestern United States.

Populus fremontii is among the most dominant, and ecologically important riparian tree species in the western United States and can thrive in hyper-arid riparian corridors. Yet, P. fremontii forests have rapidly declined over the last decade, particularly in places where temperatures sometimes exceed 50°C. We evaluated high temperature tolerance of leaf metabolism, leaf thermoregulation, and leaf hydraulic function in eight P. fremontii populations spanning a 5.3°C mean annual temperature gradient in a well-watered common garden, and at source locations throughout the lower Colorado River Basin. Two major results emerged. First, despite having an exceptionally high Tcrit (the temperature at which Photosystem II is disrupted) relative to other tree taxa, recent heat waves exceeded Tcrit , requiring evaporative leaf cooling to maintain leaf-to-air thermal safety margins. Second, in midsummer, genotypes from the warmest locations maintained lower midday leaf temperatures, a higher midday stomatal conductance, and maintained turgor pressure at lower water potentials than genotypes from more temperate locations. Taken together, results suggest that under well-watered conditions, P. fremontii can regulate leaf temperature below Tcrit along the warm edge of its distribution. Nevertheless, reduced Colorado River flows threaten to lower water tables below levels needed for evaporative cooling during episodic heat waves.

Use of thermal imaging and the photochemical reflectance index (PRI) to detect wheat response to elevated CO2 and drought.

The spectral-based photochemical reflectance index (PRI) and leaf surface temperature (Tleaf ) derived from thermal imaging are two indicative metrics of plant functioning. The relationship of PRI with radiation-use efficiency (RUE) and Tleaf with leaf transpiration could be leveraged to monitor crop photosynthesis and water use from space. Yet, it is unclear how such relationships will change under future high carbon dioxide concentrations ([CO2 ]) and drought. Here we established an [CO2 ] enrichment experiment in which three wheat genotypes were grown at ambient (400 ppm) and elevated (550 ppm) [CO2 ] and exposed to well-watered and drought conditions in two glasshouse rooms in two replicates. Leaf transpiration (Tr ) and latent heat flux (LE) were derived to assess evaporative cooling, and RUE was calculated from assimilation and radiation measurements on several dates along the season. Simultaneous hyperspectral and thermal images were taken at ~ $\unicode{x0007E}$ 1.5 m from the plants to derive PRI and the temperature difference between the leaf and its surrounding air ( ∆ $\unicode{x02206}$ Tleaf-air ). We found significant PRI and RUE and ∆ $\unicode{x02206}$ Tleaf-air and Tr correlations, with no significant differences among the genotypes. A PRI-RUE decoupling was observed under drought at ambient [CO2 ] but not at elevated [CO2 ], likely due to changes in photorespiration. For a LE range of 350 W m-2 , the ΔTleaf-air range was ~ $\unicode{x0007E}$ 10°C at ambient [CO2 ] and only ~ $\unicode{x0007E}$ 4°C at elevated [CO2 ]. Thicker leaves in plants grown at elevated [CO2 ] suggest higher leaf water content and consequently more efficient thermoregulation at high [CO2 ] conditions. In general, Tleaf was maintained closer to the ambient temperature at elevated [CO2 ], even under drought. PRI, RUE, ΔTleaf -air , and Tr decreased linearly with canopy depth, displaying a single PRI-RUE and ΔTleaf -air  Tr model through the canopy layers. Our study shows the utility of these sensing metrics in detecting wheat responses to future environmental changes.

Elevated [CO2] negatively impacts C4 photosynthesis under heat and water stress without penalizing biomass.

Elevated [CO2] (eCO2) and water stress reduce leaf stomatal conductance (gs), which may affect leaf thermoregulation during heat waves (heat stress). Two sorghum lines, with different leaf width were grown in a glasshouse at a mean day temperature of 30 °C, under different [CO2] and watering levels, and subjected to heat stress (43 °C) for 6 d at the start of the reproductive stage. We measured leaf photosynthetic and stomatal responses to light transients before harvesting the plants. Photosynthesis at growth conditions (Agrowth) and biomass accumulation were enhanced by eCO2 under control conditions. Heat stress increased gs, especially in wider leaves, and reduced the time constant of stomatal opening (kopen) at ambient [CO2] but not eCO2. However, heat stress reduced photosynthesis under water stress and eCO2 due to increased leaf temperature and reduced evaporative cooling. eCO2 prevented the reduction of biomass under both water and heat stress, possibly due to improved plant and soil water status as a result of reduced gs. Our results suggest that the response of the C4 crop sorghum to future climate conditions depends on the trade-off between low gs needed for high water use efficiency and drought tolerance, and the high gs needed for improved thermoregulation and heat tolerance under an eCO2 future.

Toxicity, physiological, and morphological alterations of Indian camphorweed (Pluchea indica) in response to excess copper.

Indian camphorweed (Pluchea indica (L.) Less.) is used as herbal tea due to the presence of volatile aromatic oils and several phytochemical compounds. The aim of this study was to assess the impact of copper (Cu) contamination on the physiology and morphology of P. indica, and the health risks associated with its consumption as tea. The cuttings of P. indica were subjected to 0 mM (control), 5 mM (low Cu), and 20 mM (excess Cu) of CuSO4 treatments for 1, 2, and 4 weeks. Thereafter, Cu contamination as well as physiological and morphological parameters were assessed. Cu accumulation was higher in the root tissues of plants (25.8 folds higher as compared to the leaves) grown under 20 mM CuSO4 for 4 weeks. This increased Cu accumulation resulted in the inhibition of root length, root fresh weight, and root dry weight. Cu concentration was found maximum (1.36 µg g-1 DW) in the leaf tissues under 20 mM Cu exposure for 4 weeks, with the highest target hazard quotient (THQ = 1.85), whereas Cu was not detected in control. Under exposure to 20 mM Cu treatment for 4 weeks, leaf greenness, maximum quantum yield of photosystem II, and photon yield of photosystem II diminished by 21.4%, 16.1%, and 22.4%, respectively, as compared to the control. Leaf temperature was increased by 2.5 °C, and the crop stress index (CSI) exceeded 0.6 when exposed to 20 mM Cu treatment for 2 and 4 weeks; however, the control had a CSI below 0.5. This led to a reduced transpiration rate and stomatal conductance. In addition, the net photosynthetic rate was also found sensitive to Cu treatment, which resulted in decreased shoot and root growth. Based on the key results, it can be suggested that P. indica herbal tea derived from the foliage of plants grown under a 5 mM Cu level (0.75 µg g-1 DW) with a target hazard quotient below one aligns with the recommended dietary intake of Cu in leafy vegetables. The study recommends choosing cuttings from plants with a small canopy as plant material in the greenhouse microclimates to validate the growth performance in the Cu-contaminated soil and simulate the natural shrub architecture and life cycle.

Iron (Fe) toxicity, uptake, translocation, and physio-morphological responses in Catharanthus roseus.

Iron (Fe) toxicity in plant species depends on the availability of Fe in the soil, uptake ability by the root system, and translocation rate to other parts of the plant. The aim of this study was to assess Fe uptake by root tissues of Catharanthus roseus, translocation rate to leaf tissues, and the impairment of plant physio-morphological characteristics. Fe uptake by the roots (~ 700 µg g-1 DW) of C. roseus was observed during the early exposure period (1 week), and translocation factor from root to shoot was fluctuated as an independent strategy. A high level of Fe content in the root tissues significantly inhibited root length and root dry weight. Under acidic pH condition, an enrichment of Fe in the shoots (~ 400 µg g-1 DW) led to increase in leaf temperature (> 2.5 °C compared to control) and crop stress index (> 0.6), resulting in stomatal closure, subsequently decreasing CO2 assimilation rate and H2O transpiration rate. An increment of CSI in Fe-stressed plants was negatively related to stomatal conductance, indicating stomatal closure with an increase in Fe in the leaf tissues. High Fe levels in the leaf tissues directly induced toxic symptoms including leaf bronzing, leaf spotting, leaf necrosis, leaf chlorosis, and leaf senescence in C. roseus plants. In summary, C. roseus was identified as a good candidate plant for Fe phytoextraction, depending on Fe bioaccumulation, therefore 50 mM Fe treatment was designated as an excess Fe to cause the growth inhibition, especially in the prolonged Fe incubation periods. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01379-5.

Gas exchange analysers exhibit large measurement error driven by internal thermal gradients.

Portable gas exchange analysers provide critical data for understanding plant-atmosphere carbon and water fluxes, and for parameterising Earth system models that forecast climate change effects and feedbacks. We characterised temperature measurement errors in the Li-Cor LI-6400XT and LI-6800, and estimated downstream errors in derived quantities, including stomatal conductance (gsw ) and leaf intercellular CO2 concentration (Ci ). The LI-6400XT exhibited air temperature errors (differences between reported air temperature and air temperature measured near the leaf) up to 7.2°C, leaf temperature errors up to 5.3°C, and relative errors in gsw and Ci that increased as temperatures departed from ambient. This caused errors in leaf-to-air temperature relationships, assimilation-temperature curves and CO2 response curves. Temperature dependencies of maximum Rubisco carboxylation rate (Vcmax ) and maximum RuBP regeneration rate (Jmax ) showed errors of 12% and 35%, respectively. These errors are likely to be idiosyncratic and may differ among machines and environmental conditions. The LI-6800 exhibited much smaller errors. Earth system model predictions may be erroneous, as much of their parametrisation data were measured on the LI-6400XT system, depending on the methods used. We make recommendations for minimising errors and correcting data in the LI-6400XT. We also recommend transitioning to the LI-6800 for future data collection.

Aerosol pollution alters the diurnal dynamics of sun and shade leaf photosynthesis through different mechanisms.

Anthropogenic aerosols have been shown to perturb CO2 exchange between the vegetation and the atmosphere. However, the climate effects of aerosols through carbon cycle feedback still have significant uncertainties. Taking advantage of the periodic fluctuations of aerosol loading in Beijing, we intensively measured the diurnal course of leaf microclimates and photosynthesis under different aerosol conditions during the growing season in 2014 and 2015. We found that increasing aerosol loadings altered the diurnal course of microclimates and thus sun and shade leaf photosynthesis. Our mechanistic photosynthesis model experiments further showed that aerosol-induced increase in sun leaf photosynthesis occurred around noon and afternoon, mainly by alleviating the depression of photosynthesis caused by high leaf temperature and leaf-air vapour pressure deficit. Meanwhile, aerosols enhanced shade leaf photosynthesis throughout the day by mitigating the light limitation within the canopy, with the highest increase occurring around noon. Overall, our study suggested that aerosol's diffuse fertilization effect, cooling effect and the accompanying low leaf-air vapour pressure deficit collectively drove the changes in the diurnal courses of sun and shade leaf photosynthesis. Our results provided an important benchmark for assessing how anthropogenic aerosols regulate ecosystem C balance under different meteorological conditions.

Tradeoffs between leaf cooling and hydraulic safety in a dominant arid land riparian tree species.

Leaf carbon gain optimization in hot environments requires balancing leaf thermoregulation with avoiding excessive water loss via transpiration and hydraulic failure. The tradeoffs between leaf thermoregulation and transpirational water loss can determine the ecological consequences of heat waves that are increasing in frequency and intensity. We evaluated leaf thermoregulation strategies in warm- (>40°C maximum summer temperature) and cool-adapted (<40°C maximum summer temperature) genotypes of the foundation tree species, Populus fremontii, using a common garden near the mid-elevational point of its distribution. We measured leaf temperatures and assessed three modes of leaf thermoregulation: leaf morphology, midday canopy stomatal conductance and stomatal sensitivity to vapour pressure deficit. Data were used to parameterize a leaf energy balance model to estimate contrasts in midday leaf temperature in warm- and cool-adapted genotypes. Warm-adapted genotypes had 39% smaller leaves and 38% higher midday stomatal conductance, reflecting a 3.8°C cooler mean leaf temperature than cool-adapted genotypes. Leaf temperatures modelled over the warmest months were on average 1.1°C cooler in warm- relative to cool-adapted genotypes. Results show that plants adapted to warm environments are predisposed to tightly regulate leaf temperatures during heat waves, potentially at an increased risk of hydraulic failure.

Coordination of stomata and vein patterns with leaf width underpins water-use efficiency in a C4 crop.

Despite its importance for crop water use and productivity, especially in drought-affected environments, the underlying mechanisms of variation in intrinsic water-use efficiency (iWUE = net photosynthesis/stomatal conductance for water vapour, gsw ) are not well understood, especially in C4 plants. Recently, we discovered that leaf width (LW) correlated negatively with iWUE and positively with gsw across several C4 grasses. Here, we confirmed these relationships within 48 field-grown genotypes differing in LW in Sorghum bicolor, a C4 crop adapted to dry and hot conditions. We measured leaf gas exchange and modelled leaf energy balance three times a day, alongside anatomical traits as potential predictors of iWUE. LW correlated negatively with iWUE and stomatal density, but positively with gsw , interveinal distance of longitudinal veins, and the percentage of stomatal aperture relative to maximum. Energy balance modelling showed that wider leaves needed to open their stomata more to generate a more negative leaf-to-air temperature difference, especially at midday when air temperatures exceeded 40°C. These results highlight the important role that LW plays in shaping iWUE through coordination of vein and stomatal traits and by affecting stomatal aperture. Therefore, LW could be used as a predictor of higher iWUE among sorghum genotypes.

Narrow safety margin in the phyllosphere during thermal extremes.

The thermal limit of ectotherms provides an estimate of vulnerability to climate change. It differs between contrasting microhabitats, consistent with thermal ecology predictions that a species' temperature sensitivity matches the microclimate it experiences. However, observed thermal limits may differ between ectotherms from the same environment, challenging this theory. We resolved this apparent paradox by showing that ectotherm activity generates microclimatic deviations large enough to account for differences in thermal limits between species from the same microhabitat. We studied upper lethal temperature, effect of feeding mode on plant gas exchange, and temperature of attacked leaves in a community of six arthropod species feeding on apple leaves. Thermal limits differed by up to 8 °C among the species. Species that caused an increase in leaf transpiration (+182%), thus cooling the leaf, had a lower thermal limit than those that decreased leaf transpiration (-75%), causing the leaf to warm up. Therefore, cryptic microclimatic variations at the scale of a single leaf determine the thermal limit in this community of herbivores. We investigated the consequences of these changes in plant transpiration induced by plant-insect feedbacks for species vulnerability to thermal extremes. Warming tolerance was similar between species, at ±2 °C, providing little margin for resisting increasingly frequent and intense heat waves. The thermal safety margin (the difference between thermal limit and temperature) was greatly overestimated when air temperature or intact leaf temperature was erroneously used. We conclude that feedback processes define the vulnerability of species in the phyllosphere, and beyond, to thermal extremes.