A bitter future for coffee production? Physiological traits associated with yield reveal high vulnerability to hydraulic failure in Coffea canephora.

The increase in frequency and intensity of drought events have hampered coffee production in the already threatened Amazon region, yet little is known about key aspects underlying the variability in yield potential across genotypes, nor to what extent higher productivity is linked to reduced drought tolerance. Here we explored how variations in morphoanatomical and physiological leaf traits can explain differences in yield and vulnerability to embolism in 11 Coffea canephora genotypes cultivated in the Western Amazon. The remarkable variation in coffee yield across genotypes was tightly related to differences in their carbon assimilation and water transport capacities, revealing a diffusive limitation to photosynthesis linked by hydraulic constraints. Although a clear trade-off between water transport efficiency and safety was not detected, all the studied genotypes operated in a narrow and/or negative hydraulic safety margin, suggesting a high vulnerability to leaf hydraulic failure (HF), especially on the most productive genotypes. Modelling exercises revealed that variations in HF across genotypes were mainly associated with differences in leaf water vapour leakage when stomata are closed, reflecting contrasting growth strategies. Overall, our results provide a new perspective on the challenges of sustaining coffee production in the Amazon region under a drier and warmer climate.

Leaf habit affects the distribution of drought sensitivity but not water transport efficiency in the tropics.

Considering the global intensification of aridity in tropical biomes due to climate change, we need to understand what shapes the distribution of drought sensitivity in tropical plants. We conducted a pantropical data synthesis representing 1117 species to test whether xylem-specific hydraulic conductivity (KS ), water potential at leaf turgor loss (ΨTLP ) and water potential at 50% loss of KS (ΨP50 ) varied along climate gradients. The ΨTLP and ΨP50 increased with climatic moisture only for evergreen species, but KS did not. Species with high ΨTLP and ΨP50 values were associated with both dry and wet environments. However, drought-deciduous species showed high ΨTLP and ΨP50 values regardless of water availability, whereas evergreen species only in wet environments. All three traits showed a weak phylogenetic signal and a short half-life. These results suggest strong environmental controls on trait variance, which in turn is modulated by leaf habit along climatic moisture gradients in the tropics.

Hydraulic traits vary as the result of tip-to-base conduit widening in vascular plants.

Plant hydraulic traits are essential metrics for characterizing variation in plant function, but they vary markedly with plant size and position in a plant. We explore the potential effect of conduit widening on variation in hydraulic traits along the stem. We examined three species that differ in conduit diameter at the stem base for a given height (Moringa oleifera, Casimiroa edulis, and Pinus ayacahuite). We made anatomical and hydraulic measurements at different distances from the stem tip, constructed vulnerability curves, and examined the safety-efficiency trade-off with height-standardized data. Our results showed that segment-specific hydraulic resistance varied predictably along the stem, paralleling changes in mean conduit diameter and total number of conduits. The Huber value and leaf specific conductivity also varied depending on the sampling point. Vulnerability curves were markedly less noisy with height standardization, making the vulnerability-efficiency trade-off clearer. Because conduits widen predictably along the stem, taking height and distance from the tip into account provides a way of enhancing comparability and interpretation of hydraulic traits. Our results suggest the need for rethinking hydraulic sampling for comparing plant functional differences and strategies across individuals.

From Carlquist's ecological wood anatomy to Carlquist's Law: why comparative anatomy is crucial for functional xylem biology.

All students of xylem structure-function relations need to be familiar with the work of Sherwin Carlquist. He studies xylem through the lens of the comparative method, which uses the appearance of similar anatomical features under similar conditions of natural selection to infer function. "Function" in biology implies adaptation; maximally supported adaptation inferences require experimental and comparative xylem scientists to work with one another. Engaging with comparative inferences of xylem function will, more likely sooner rather than later, bring one to the work of Sherwin Carlquist. To mark his 90th birthday, I highlight just a few examples of his extraordinarily perceptive and general comparative insights. One is "Carlquist's Law", the pervasive tendency for vessels to be solitary when background cells are conductive. I cover his pioneering of "ecological" wood anatomy, viewing xylem variation as reflecting the effects of selection across climate and habit variation. Another is the embolism vulnerability-conduit diameter relationship, one of the most widely invoked structure-function relationships in xylem biology. I discuss the inferential richness within the notion of Carlquistian paedomorphosis, including detailed functional inferences regarding ray cell orientation. My final example comes from his very recent work offering the first satisfactory hypothesis accounting for the geographical and histological distribution of scalariform perforation plates as an adaptation, including "Carlquist's Ratchet", why scalariform plates are adaptive but do not re-evolve once lost. This extraordinarily rich production over six decades is filled with comparative inferences that should keep students of xylem function busy testing for decades to come.

No local adaptation in leaf or stem xylem vulnerability to embolism, but consistent vulnerability segmentation in a North American oak.

Vulnerability to embolism varies between con-generic species distributed along aridity gradients, yet little is known about intraspecific variation and its drivers. Even less is known about intraspecific variation in tissues other than stems, despite results suggesting that roots, stems and leaves can differ in vulnerability. We hypothesized that intraspecific variation in vulnerability in leaves and stems is adaptive and driven by aridity. We quantified leaf and stem vulnerability of Quercus douglasii using the optical technique. To assess contributions of genetic variation and phenotypic plasticity to within-species variation, we quantified the vulnerability of individuals growing in a common garden, but originating from populations along an aridity gradient, as well as individuals from the same wild populations. Intraspecific variation in water potential at which 50% of total embolism in a tissue is observed (P50 ) was explained mostly by differences between individuals (>66% of total variance) and tissues (16%). There was little between-population variation in leaf/stem P50 in the garden, which was not related to site of origin aridity. Unexpectedly, we observed a positive relationship between wild individual stem P50 and aridity. Although there is no local adaptation and only minor phenotypic plasticity in leaf/stem vulnerability in Q. douglasii, high levels of potentially heritable variation within populations or strong environmental selection could contribute to adaptive responses under future climate change.

A dynamic yet vulnerable pipeline: Integration and coordination of hydraulic traits across whole plants.

The vast majority of measurements in the field of plant hydraulics have been on small-diameter branches from woody species. These measurements have provided considerable insight into plant functioning, but our understanding of plant physiology and ecology would benefit from a broader view, because branch hydraulic properties are influenced by many factors. Here, we discuss the influence that other components of the hydraulic network have on branch vulnerability to embolism propagation. We also modelled the impact of changes in the ratio of root-to-leaf areas and soil texture on vulnerability to hydraulic failure along the soil-to-leaf continuum and showed that hydraulic function is better maintained through changes in root vulnerability and root-to-leaf area ratio than in branch vulnerability. Differences among species in the stringency with which they regulate leaf water potential and in reliance on stored water to buffer changes in water potential also affect the need to construct embolism resistant branches. Many approaches, such as measurements on fine roots, small individuals, combining sap flow and psychrometry techniques, and modelling efforts, could vastly improve our understanding of whole-plant hydraulic functioning. A better understanding of how traits are coordinated across the whole plant will improve predictions for plant function under future climate conditions.

Post-fire effects in xylem hydraulics of Picea abies, Pinus sylvestris and Fagus sylvatica.

Recent studies on post-fire tree mortality suggest a role for heat-induced alterations of the hydraulic system. We analyzed heat effects on xylem hydraulics both in the laboratory and at a forest site hit by fire. Stem vulnerability to drought-induced embolism and hydraulic conductivity were measured in Picea abies, Pinus sylvestris and Fagus sylvatica. Control branches were compared with samples experimentally exposed to 90°C or damaged by a natural forest fire. In addition, xylem anatomical changes were examined microscopically. Experimental heating caused structural changes in the xylem and increased vulnerability in all species. The largest shifts in vulnerability thresholds (1.3 MPa) were observed in P. sylvestris. F. sylvatica also showed heat-induced reductions (49%) in hydraulic conductivity. At the field site, increased vulnerability was observed in damaged branches of P. sylvestris and F. sylvatica, and the xylem of F. sylvatica was 39% less conductive in damaged than in undamaged branches. These results provide evidence for heat-induced impairment of tree hydraulics after fire. The effects recorded at the forest fire site corresponded to those obtained in laboratory experiments, and revealed pronounced hydraulic risks in P. sylvestris and F. sylvatica. Knowledge of species-specific hydraulic impairments induced by fire and heat is a prerequisite for accurate estimation of post-fire mortality risks.

Direct observation and modelling of embolism spread between xylem conduits: a case study in Scots pine.

Xylem embolism is one of the main processes involved in drought-related plant mortality. Although its consequences for plant physiology are already well described, embolism formation and spread are poorly evaluated and modelled, especially for tracheid-based species. The aim of this study was to assess the embolism formation and spread in Pinus sylvestris as a case study using X-ray microtomography and hydraulics methods. We also evaluated the potential effects of cavitation fatigue on vulnerability to embolism and the micro-morphology of the bordered pits using scanning electron microscopy (SEM) to test for possible links between xylem anatomy and embolism spread. Finally, a novel model was developed to simulate the spread of embolism in a 2D anisotropic cellular structure. Results showed a large variability in the formation and spread of embolism within a ring despite no differences being observed in intertracheid pit membrane anatomical traits. Simulations from the model showed a highly anisotropic tracheid-to-tracheid embolism spreading pattern, which confirms the major role of tracheid-to-tracheid air seeding to explain how embolism spreads in Scots pine. The results also showed that prior embolism removal from the samples reduced the resistance to embolism of the xylem and could result in overestimates of vulnerability to embolism.

Static and dynamic bending has minor effects on xylem hydraulics of conifer branches (Picea abies, Pinus sylvestris).

The xylem hydraulic efficiency and safety is usually measured on mechanically unstressed samples, although trees may be exposed to combined hydraulic and mechanical stress in the field. We analysed changes in hydraulic conductivity and vulnerability to drought-induced embolism during static bending of Picea abies and Pinus sylvestris branches as well as the effect of dynamic bending on the vulnerability. We hypothesized this mechanical stress to substantially impair xylem hydraulics. Intense static bending caused an only small decrease in hydraulic conductance (-19.5 ± 2.4% in P. abies) but no shift in vulnerability thresholds. Dynamic bending caused a 0.4 and 0.8 MPa decrease of the water potential at 50 and 88% loss of conductivity in P. sylvestris, but did not affect vulnerability thresholds in P. abies. With respect to applied extreme bending radii, effects on plant hydraulics were surprisingly small and are thus probably of minor eco-physiological importance. More importantly, results indicate that available xylem hydraulic analyses (of conifers) sufficiently reflect plant hydraulics under field conditions.

Lack of phenotypic plasticity in leaf hydraulics for 10 woody species common to urban forests of North China.

The survival and performance of urban forests are increasingly challenged by urban drought, consequently compromising the sustainability and functionality of urban vegetation. Plant-water relations largely determine species drought tolerance, yet little is known about the hydraulics of urban forest species. Here, we report the leaf hydraulic and carbon traits that govern plant growth and drought resistance, including vulnerability to embolism, hydraulic conductivity and leaf gas exchange characteristics, as well as morphological traits that are potentially linked with these physiological attributes, with the aim of guiding species selection and management in urban forests. Plant materials were collected from mature shrubs and trees on our university campus in Beijing, representing 10 woody species common to urban forests in north China. We found that the leaf embolism resistance, represented by the water potential inducing 50% loss of hydraulic conductivity (P50), as well as the hydraulic safety margin (HSM) defined by P50 and the water potential threshold at the inception of embolism (P12), varied remarkably across species, but was unrelated to growth form. Likewise, stem and leaf-specific hydraulic conductivity (Kstem and kl) was also highly species-specific. Leaf P50 was positively correlated with hydraulic conductivity. However, neither P50 nor hydraulic conductivity was correlated with leaf gas exchange traits, including maximum photosynthetic rate (Amax) and stomatal conductance (gs). Plant morphological and physiological traits were not related, except for specific leaf area, which showed a negative relationship with HSM. Traits influencing plant-water transport were primarily correlated with the mean annual precipitation of species climatic niche. Overall, current common woody species in urban forest environments differed widely in their drought resistance and did not have the capacity to modify these characteristics in response to a changing climate. Species morphology provides limited information regarding physiological drought resistance. Thus, screening urban forest species based on plant physiology is essential to sustain the ecological services of urban forests.

A User Manual to Measure Gas Diffusion Kinetics in Plants: Pneumatron Construction, Operation, and Data Analysis.

The Pneumatron device measures gas diffusion kinetics in the xylem of plants. The device provides an easy, low-cost, and powerful tool for research on plant water relations and gas exchange. Here, we describe in detail how to construct and operate this device to estimate embolism resistance of angiosperm xylem, and how to analyse pneumatic data. Simple and more elaborated ways of constructing a Pneumatron are shown, either using wires, a breadboard, or a printed circuit board. The instrument is based on an open-source hardware and software system, which allows users to operate it in an automated or semi-automated way. A step-by-step manual and a troubleshooting section are provided. An excel spreadsheet and an R-script are also presented for fast and easy data analysis. This manual aims at helping users to avoid common mistakes, such as unstable measurements of the minimum and maximum amount of gas discharged from xylem tissue, which has major consequences for estimating embolism resistance. Major advantages of the Pneumatron device include its automated and accurate measurements of gas diffusion rates, including highly precise measurements of the gas volume in intact, embolised conduits. It is currently unclear if the method can also be applied to woody monocots, gymnosperm species that possess torus-margo pit membranes, or to herbaceous species.

The potential of Mid-Infrared spectroscopy for prediction of wood density and vulnerability to embolism in woody angiosperms.

Xylem resistance to embolism formation determines the species-specific drought tolerance and the survival prospects of plants under extreme climatic conditions. Fourier Transform-Infrared (FTIR) spectroscopy is a cost-effective and rapid analytical tool with potential beyond its current use in plant physiology. We tested the use of FTIR spectroscopy as a method for estimating wood density (WD) and xylem resistance to embolism formation (P50) in 24 angiosperm species. Higher WD was associated with more negative P50 (r2 = 0.41). Partial least squares regression was applied to establish models of FTIR spectra and the reference data. They showed a high predictive quality for WD (r2 = 0.73), whereas the prediction of P50 was weaker (r2 = 0.49). By including WD in the model as an additional factor influencing P50, its predictive power significantly increased (r2 = 0.59). The spectral range in the model elaboration has been also narrowed (bands of lignin, cellulose, hemicellulose), but this did not influence the model descriptors, suggesting that for P50 prediction broad spectral range is more informative than narrow band regions reflecting main wood constituents. In conclusion, FTIR spectroscopy associated with WD measurements has proven to be a promising alternative to traditional methods for screening of individual- or species-specific resistance to embolism in angiosperms.

A comparison of hydraulic architecture in three similarly sized woody species differing in their maximum potential height.

The physiological mechanisms underlying the short maximum height of shrubs are not understood. One possible explanation is that differences in the hydraulic architecture of shrubs compared with co-occurring taller trees prevent the shrubs from growing taller. To explore this hypothesis, we examined various hydraulic parameters, including vessel lumen diameter, hydraulic conductivity and vulnerability to drought-induced embolism, of three co-occurring species that differed in their maximum potential height. We examined one species of shrub, one short-statured tree and one taller tree. We worked with individuals that were approximately the same age and height, which was near the maximum for the shrub species. A number of variables correlated with the maximum potential height of the species. For example, vessel diameter and vulnerability to embolism both increased while wood density declined with maximum potential height. The difference between the pressure causing 50% reduction in hydraulic conductance in the leaves and the midday leaf water potential (the leaf's hydraulic safety margin) was much larger in the shrub than the other two species. In general, trends were consistent with understory shrubs having a more conservative life history strategy than co-occurring taller species.