Sap flow through partially embolized xylem vessel networks.

Sap is transported through numerous conduits in the xylem of woody plants along the path from the soil to the leaves. When all conduits are functional, vessel lumen diameter is a strong predictor of hydraulic conductivity. As vessels become embolized, sap movement becomes increasingly affected by factors operating at scales beyond individual conduits, creating resistances that result in hydraulic conductivity diverging from diameter-based estimates. These effects include pit resistances, connectivity, path length, network topology, and vessel or sector isolation. The impact of these factors varies with the level and distribution of emboli within the network, and manifest as alterations in the relationship between the number and diameter of embolized vessels with measured declines in hydraulic conductivity across vulnerability to embolism curves. Divergences between measured conductivity and diameter-based estimates reveal functional differences that arise because of species- and tissue-specific vessel network structures. Such divergences are not uniform, and xylem tissues may diverge in different ways and to differing degrees. Plants regularly operate under nonoptimal conditions and contain numerous embolized conduits. Understanding the hydraulic implications of emboli within a network and the function of partially embolized networks are critical gaps in our understanding of plants occurring within natural environments.

Foliar water uptake in Pinus species depends on needle age and stomatal wax structures.

BACKGROUND AND AIMS: Foliar water uptake (FWU) has been documented in many species and is increasingly recognized as a non-trivial factor in plant-water relationships. However, it remains unknown whether FWU is a widespread phenomenon in Pinus species, and how it may relate to needle traits such as the form and structure of stomatal wax plugs. In this contribution, these questions were addressed by studying FWU in current-year and 1-year-old needles of seven Pinus species. METHODS: We monitored FWU gravimetrically and analysed the needle surface via cryo-scanning electron microscopy. Additionally, we considered the effect of artificial wax erosion by application of the surfactant Triton X-100, which is able to alter wax crystals. KEY RESULTS: The results show for all species that (1) FWU occurred, (2) FWU is higher in old needles compared to young needles and (3) there is substantial erosion of stomatal wax plugs in old needles. FWU was highest in Pinus canariensis, which has a thin stomatal wax plug. Surfactant treatment enhanced FWU. CONCLUSIONS: The results of this study provide evidence for (1) widespread FWU in Pinus, (2) the influence of stomatal wax plugs on FWU and (3) age-related needle surface erosion.

Solid mechanics of the torus-margo in conifer intertracheid bordered pits.

Bordered pits of many conifers include a torus-margo structure acting as a valve that prevents air from spreading between tracheids, although the extent of torus deflection as a function of applied pressure is not well known. Models were developed from images of pits in roots and stems of Picea mariana (Mill.) BSP. A computational solid mechanics approach was utilised to determine the extent of torus deflection from pressure applied to the torus and margo. Torus deflection increased in nonlinear fashion with applied pressure. The average pressure required for sealing the pit was 0.894 MPa for stems and 0.644 MPa for roots, although considerable variation was apparent between individual pits. The pits of roots were wider and deeper than those of stems. For stems, the pit depth did not increase with pit width; thus the torus displacement needed to seal the pit was less than for pits from roots. The pressure required to seal the pit depends upon anatomical characteristics such as pit width and pit depth. Although the torus displacement for sealing was greater for roots because of their greater depth, the pressures leading to sealing were not significantly different between roots and stems.

Seasonal patterns of callose deposition and xylem embolism in five boreal deciduous tree species.

PREMISE: Phloem tissue allows for sugar transport along the entirety of a plant and, thus, is one of the most important anatomical structures related to growth. It is thought that the sugar-conducting sieve tube may overwinter and that its cells persist multiple seasons in deciduous trees. One possible overwintering strategy is to build up callose on phloem sieve plates to temporarily cease their function. We tested the hypothesis that five deciduous tree species produce callose on their sieve plates on a seasonal basis. METHODS: Young shoots of five deciduous tree species were sampled periodically between April 2019 and February 2020 in Edmonton, Alberta, Canada. After enzymatic digestion of cytoplasmic constituents, cross sections were imaged using scanning electron microscopy to observe and quantify the level of callose deposition at monthly intervals, and sieve plate pore size was measured. Using a conductivity apparatus, we measured xylem native embolism during these sampling periods. RESULTS: Contrary to past work on some of the same species, we found little evidence that sieve tubes overwinter by becoming occluded with callose. Instead, we found that most sieve plates remain open. Xylem embolism was minimal during the peak growing season, but increased over winter. CONCLUSIONS: Many species had been assumed to deposit callose on sieve plates over winter, though anatomical and phenological phloem data were sparse. Our data do not support this notion.

Computational models evaluating the impact of sieve plates and radial water exchange on phloem pressure gradients.

The sugar conducting phloem in angiosperms is a high resistance pathway made up of sieve elements bounded by sieve plates. The high resistance generated by sieve plates may be a trade-off for promoting quick sealing in the event of injury. However, previous modeling efforts have demonstrated a wide variation in the contribution of sieve plates towards total sieve tube resistance. In the current study, we generated high resolution scanning electron microscope images of sieve plates from balsam poplar and integrated them into a mathematical model using Comsol Multiphysics software. We found that sieve plates contribute upwards of 85% towards total sieve tube resistance. Utilizing the Navier-Stokes equations, we found that oblong pores may create over 50% more resistance in comparison with round pores of the same area. Although radial water flows in phloem sieve tubes have been previously considered, their impact on alleviating pressure gradients has not been fully studied. Our novel simulations find that radial water flow can reduce pressure requirements by half in comparison with modeled sieve tubes with no radial permeability. We discuss the implication that sieve tubes may alleviate pressure requirements to overcome high resistances by regulating their membrane permeability along the entire transport pathway.

Direct comparison of four methods to construct xylem vulnerability curves: Differences among techniques are linked to vessel network characteristics.

During periods of dehydration, water transport through xylem conduits can become blocked by embolism formation. Xylem embolism compromises water supply to leaves and may lead to losses in productivity or plant death. Vulnerability curves (VCs) characterize plant losses in conductivity as xylem pressures decrease. VCs are widely used to characterize and predict plant water use at different levels of water availability. Several methodologies for constructing VCs exist and sometimes produce different results for the same plant material. We directly compared four VC construction methods on stems of black cottonwood (Populus trichocarpa), a model tree species: dehydration, centrifuge, X-ray-computed microtomography (microCT), and optical. MicroCT VC was the most resistant, dehydration and centrifuge VCs were intermediate, and optical VC was the most vulnerable. Differences among VCs were not associated with how cavitation was induced but were related to how losses in conductivity were evaluated: measured hydraulically (dehydration and centrifuge) versus evaluated from visual information (microCT and optical). Understanding how and why methods differ in estimating vulnerability to xylem embolism is important for advancing knowledge in plant ecophysiology, interpreting literature data, and using accurate VCs in water flux models for predicting plant responses to drought.

Global convergence in the vulnerability of forests to drought.

Shifts in rainfall patterns and increasing temperatures associated with climate change are likely to cause widespread forest decline in regions where droughts are predicted to increase in duration and severity. One primary cause of productivity loss and plant mortality during drought is hydraulic failure. Drought stress creates trapped gas emboli in the water transport system, which reduces the ability of plants to supply water to leaves for photosynthetic gas exchange and can ultimately result in desiccation and mortality. At present we lack a clear picture of how thresholds to hydraulic failure vary across a broad range of species and environments, despite many individual experiments. Here we draw together published and unpublished data on the vulnerability of the transport system to drought-induced embolism for a large number of woody species, with a view to examining the likely consequences of climate change for forest biomes. We show that 70% of 226 forest species from 81 sites worldwide operate with narrow (<1 megapascal) hydraulic safety margins against injurious levels of drought stress and therefore potentially face long-term reductions in productivity and survival if temperature and aridity increase as predicted for many regions across the globe. Safety margins are largely independent of mean annual precipitation, showing that there is global convergence in the vulnerability of forests to drought, with all forest biomes equally vulnerable to hydraulic failure regardless of their current rainfall environment. These findings provide insight into why drought-induced forest decline is occurring not only in arid regions but also in wet forests not normally considered at drought risk.

An ecophysiological and developmental perspective on variation in vessel diameter.

Variation in xylem vessel diameter is one of the most important parameters when evaluating plant water relations. This review provides a synthesis of the ecophysiological implications of variation in lumen diameter together with a summary of our current understanding of vessel development and its endogenous regulation. We analyzed inter-specific variation of the mean hydraulic vessel diameter (Dv ) across biomes, intra-specific variation of Dv under natural and controlled conditions, and intra-plant variation. We found that the Dv measured in young branches tends to stay below 30 µm in regions experiencing winter frost, whereas it is highly variable in the tropical rainforest. Within a plant, the widest vessels are often found in the trunk and in large roots; smaller diameters have been reported for leaves and small lateral roots. Dv varies in response to environmental factors and is not only a function of plant size. Despite the wealth of data on vessel diameter variation, the regulation of diameter is poorly understood. Polar auxin transport through the vascular cambium is a key regulator linking foliar and xylem development. Limited evidence suggests that auxin transport is also a determinant of vessel diameter. The role of auxin in cell expansion and in establishing longitudinal continuity during secondary growth deserve further study.

Are phloem sieve tubes leaky conduits supported by numerous aquaporins?

PREMISE OF THE STUDY: Aquaporin membrane water channels have been previously identified in the phloem of angiosperms, but currently their cellular characterization is lacking, especially in tree species. Pinpointing the cellular location will help generate new hypotheses of how membrane water exchange facilitates sugar transport in plants. METHODS: We studied histological sections of balsam poplar (Populus balsamifera L.) in leaf, petiole, and stem organs. Immuno-labeling techniques were used to characterize the distribution of PIP1 and PIP2 subfamilies of aquaporins along the phloem pathway. Confocal and super resolution microscopy (3D-SIM) was used to identify the localization of aquaporins at the cellular level. KEY RESULTS: Sieve tubes of the leaf lamina, petiole, and stem were labeled with antibodies directed at PIP1s and PIP2s. While PIP2s were mostly observed in the plasma membrane, PIP1s showed both an internal membrane and plasma membrane labeling pattern. CONCLUSIONS: The specificity and consistency of PIP2 labeling in sieve element plasma membranes points to high water exchange rates between sieve tubes and adjacent cells. The PIP1s may relocate between internal membranes and the plasma membrane to facilitate dynamic changes in membrane permeability of sieve elements in response to changing internal or environmental conditions. Aquaporin-mediated changes in membrane permeability of sieve tubes would also allow for some control of radial exchange of water between xylem and phloem.

Plant xylem hydraulics: What we understand, current research, and future challenges.

Herein we review the current state-of-the-art of plant hydraulics in the context of plant physiology, ecology, and evolution, focusing on current and future research opportunities. We explain the physics of water transport in plants and the limits of this transport system, highlighting the relationships between xylem structure and function. We describe the great variety of techniques existing for evaluating xylem resistance to cavitation. We address several methodological issues and their connection with current debates on conduit refilling and exponentially shaped vulnerability curves. We analyze the trade-offs existing between water transport safety and efficiency. We also stress how little information is available on molecular biology of cavitation and the potential role of aquaporins in conduit refilling. Finally, we draw attention to how plant hydraulic traits can be used for modeling stomatal responses to environmental variables and climate change, including drought mortality.

Weak tradeoff between xylem safety and xylem-specific hydraulic efficiency across the world's woody plant species.

The evolution of lignified xylem allowed for the efficient transport of water under tension, but also exposed the vascular network to the risk of gas emboli and the spread of gas between xylem conduits, thus impeding sap transport to the leaves. A well-known hypothesis proposes that the safety of xylem (its ability to resist embolism formation and spread) should trade off against xylem efficiency (its capacity to transport water). We tested this safety-efficiency hypothesis in branch xylem across 335 angiosperm and 89 gymnosperm species. Safety was considered at three levels: the xylem water potentials where 12%, 50% and 88% of maximal conductivity are lost. Although correlations between safety and efficiency were weak (r(2)  < 0.086), no species had high efficiency and high safety, supporting the idea for a safety-efficiency tradeoff. However, many species had low efficiency and low safety. Species with low efficiency and low safety were weakly associated (r(2)  < 0.02 in most cases) with higher wood density, lower leaf- to sapwood-area and shorter stature. There appears to be no persuasive explanation for the considerable number of species with both low efficiency and low safety. These species represent a real challenge for understanding the evolution of xylem.

Xylem refilling - a question of sugar transporters and pH?

This article comments on: Accumulation of sugars in the xylem apoplast observed under water stress conditions is controlled by xylem pH.

Drought-induced xylem pit membrane damage in aspen and balsam poplar.

Drought induces an increase in a tree's vulnerability to a loss of its hydraulic conductivity in many tree species, including two common in western Canada, trembling aspen (Populus tremuloides) and balsam poplar (Populus balsamifera). Termed 'cavitation fatigue' or 'air-seeding fatigue', the mechanism of this phenomenon is not well understood, but hypothesized to be a result of damage to xylem pit membranes. To examine the validity of this hypothesis, the effect of drought on the porosity of pit membranes in aspen and balsam poplar was investigated. Controlled drought and bench dehydration treatments were used to induce fatigue and scanning electron microscopy (SEM) was used to image pit membranes for relative porosity evaluations from air-dried samples after ethanol dehydration. A significant increase in the diameter of the largest pore was found in the drought and dehydration treatments of aspen, while an increase in the percentage of porous pit membranes was found in the dehydration treatments of both species. Additionally, the location of the largest pore per pit membrane was observed to tend toward the periphery of the membrane.

Leaf size serves as a proxy for xylem vulnerability to cavitation in plantation trees.

Hybrid poplars are an important renewable forest resource known for their high productivity. At the same time, they are highly vulnerable to water stress. Identifying traits that can serve as indicators for growth performance remains an important task, particularly under field conditions. Understanding which trait combinations translate to improved productivity is key in order to satisfy the demand for poplar wood in an uncertain future climate. In this study, we compared hydraulic and leaf traits among five hybrid poplar clones at 10 plantations in central Alberta. We also assessed the variation of these traits between 2- to 3-year-old branches from the lower to mid-crown and current-year long shoots from the mid to upper crown. Our results showed that (1) hybrid poplars differed in key hydraulic parameters between branch type, (2) variation of hydraulic traits among clones was relatively large for some clones and less for others, and (3) strong relationships between measured hydraulic traits, such as vessel diameter, cavitation resistance, xylem-specific and leaf-specific conductivity and leaf area, were observed. Our results suggest that leaf size could serve as an additional screening tool when selecting for drought-tolerant genotypes in forest management and tree improvement programmes.

Plant hydraulics as a central hub integrating plant and ecosystem function: meeting report for 'Emerging Frontiers in Plant Hydraulics' (Washington, DC, May 2015).

Water plays a central role in plant biology and the efficiency of water transport throughout the plant affects both photosynthetic rate and growth, an influence that scales up deterministically to the productivity of terrestrial ecosystems. Moreover, hydraulic traits mediate the ways in which plants interact with their abiotic and biotic environment. At landscape to global scale, plant hydraulic traits are important in describing the function of ecological communities and ecosystems. Plant hydraulics is increasingly recognized as a central hub within a network by which plant biology is connected to palaeobiology, agronomy, climatology, forestry, community and ecosystem ecology and earth-system science. Such grand challenges as anticipating and mitigating the impacts of climate change, and improving the security and sustainability of our food supply rely on our fundamental knowledge of how water behaves in the cells, tissues, organs, bodies and diverse communities of plants. A workshop, 'Emerging Frontiers in Plant Hydraulics' supported by the National Science Foundation, was held in Washington DC, 2015 to promote open discussion of new ideas, controversies regarding measurements and analyses, and especially, the potential for expansion of up-scaled and down-scaled inter-disciplinary research, and the strengthening of connections between plant hydraulic research, allied fields and global modelling efforts.

Pit membrane structure is highly variable and accounts for a major resistance to water flow through tracheid pits in stems and roots of two boreal conifer species.

The flow of xylem sap in conifers is strongly dependent on the presence of a low resistance path through bordered pits, particularly through the pores present in the margo of the pit membrane. A computational fluid dynamics approach was taken, solving the Navier-Stokes equation for models based on the geometry of pits observed in tracheids from stems and roots of Picea mariana (black spruce) and Picea glauca (white spruce). Model solutions demonstrate a close, inverse relationship between the total resistance of bordered pits and the total area of margo pores. Flow through the margo was dominated by a small number of the widest pores. Particularly for pits where the margo component of flow resistance was low relative to that of the torus, pore location near the inner edge of the margo allowed for greater flow than that occurring through similar-sized pores near the outer edge of the margo. Results indicate a surprisingly large variation in pit structure and flow characteristics. Nonetheless, pits in roots have lower resistance to flow than those in stems because the pits were wider and consisted of a margo with a larger area in pores.

The standard centrifuge method accurately measures vulnerability curves of long-vesselled olive stems.

The standard centrifuge method has been frequently used to measure vulnerability to xylem cavitation. This method has recently been questioned. It was hypothesized that open vessels lead to exponential vulnerability curves, which were thought to be indicative of measurement artifact. We tested this hypothesis in stems of olive (Olea europea) because its long vessels were recently claimed to produce a centrifuge artifact. We evaluated three predictions that followed from the open vessel artifact hypothesis: shorter stems, with more open vessels, would be more vulnerable than longer stems; standard centrifuge-based curves would be more vulnerable than dehydration-based curves; and open vessels would cause an exponential shape of centrifuge-based curves. Experimental evidence did not support these predictions. Centrifuge curves did not vary when the proportion of open vessels was altered. Centrifuge and dehydration curves were similar. At highly negative xylem pressure, centrifuge-based curves slightly overestimated vulnerability compared to the dehydration curve. This divergence was eliminated by centrifuging each stem only once. The standard centrifuge method produced accurate curves of samples containing open vessels, supporting the validity of this technique and confirming its utility in understanding plant hydraulics. Seven recommendations for avoiding artefacts and standardizing vulnerability curve methodology are provided.

Uptake of water via branches helps timberline conifers refill embolized xylem in late winter.

Xylem embolism is a limiting factor for woody species worldwide. Conifers at the alpine timberline are exposed to drought and freeze-thaw stress during winter, which induce potentially lethal embolism. Previous studies indicated that timberline trees survive by xylem refilling. In this study on Picea abies, refilling was monitored during winter and spring seasons and analyzed in the laboratory and in situ experiments, based on hydraulic, anatomical, and histochemical methods. Refilling started in late winter, when the soil was frozen and soil water not available for the trees. Xylem embolism caused up to 86.2% ± 3.1% loss of conductivity and was correlated with the ratio of closed pits. Refilling of xylem as well as recovery in shoot conductance started in February and corresponded with starch accumulation in secondary phloem and in the mesophyll of needles, where we also observed increasing aquaporin densities in the phloem and endodermis. This indicates that active, cellular processes play a role for refilling even under winter conditions. As demonstrated by our experiments, water for refilling was thereby taken up via the branches, likely by foliar water uptake. Our results suggest that refilling is based on water shifts to embolized tracheids via intact xylem, phloem, and parenchyma, whereby aquaporins reduce resistances along the symplastic pathway and aspirated pits facilitate isolation of refilling tracheids. Refilling must be taken into account as a key process in plant hydraulics and in estimating future effects of climate change on forests and alpine tree ecosystems.

Exploring Picea glauca aquaporins in the context of needle water uptake and xylem refilling.

Conifer needles have been reported to absorb water under certain conditions. Radial water movement across needle tissues is likely influenced by aquaporin (AQP) water channels. Foliar water uptake and AQP localization in Picea glauca needles were studied using physiological and microscopic methods. AQP expression was measured using quantitative real-time PCR. Members of the AQP gene family in spruce were identified using homology search tools. Needles of drought-stressed plants absorbed water when exposed to high relative humidity (RH). AQPs were present in the endodermis-like bundle sheath, in phloem cells and in the transfusion parenchyma of needles. Up-regulation of AQPs in high RH coincided with embolism repair in stem xylem. The present study also provides the most comprehensive functional and phylogenetic analysis of spruce AQPs to date. Thirty putative complete AQP sequences were found. Our findings are consistent with the hypothesis that AQPs facilitate radial water movement from the needle epidermis towards the vascular tissue. Foliar water uptake may occur in late winter when needles are covered by melting snow and may provide a water source for embolism repair before the beginning of the growing season.

Sixteen years of winter stress: an assessment of cold hardiness, growth performance and survival of hybrid poplar clones at a boreal planting site.

In recent years, thousands of hectares of hybrid poplar plantations have been established in Canada for the purpose of carbon sequestration and wood production. However, boreal planting environments pose special challenges that may compromise the long-term survival and productivity of such plantations. In this study, we evaluated the effect of winter stress, that is, frequent freeze-thaw and extreme cold events, on growth and survival of 47 hybrid poplar clones in a long-term field experiment. We further assessed physiological and structural traits that are potentially important for cold tolerance for a selected set of seven clones. We found that trees with narrow xylem vessels showed reduced freezing-induced embolism and showed superior productivity after 16 growing seasons. With respect to cold hardiness of living tissues, we only observed small differences among clones in early autumn, which were nonetheless significantly correlated to growth. Maximum winter cold hardiness and the timing of leaf senescence and budbreak were not related to growth or survival. In conclusion, our data suggest that reduction of freezing-induced embolism due to small vessel diameters is an essential adaptive trait to ensure long-term productivity of hybrid poplar plantations in boreal planting environments.