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In plants, xylem vessel length is important for long-distance water transport; however, the currently used methods for vessel length measurement are inconvenient and time-consuming. The recently developed semi-automated Pneumatron is a device based on the pneumatic theory that is similar to the air-injection method, and can rapidly estimate vessel length. Mean vessel length was compared between the Pneumatron and the air-injection method in seven woody species with a wide range of vessel lengths (2.3–78.7 cm). The results were consistent between the two methods, regardless of whether the same or different samples were used. The theory underlying the gas flow in vessels was improved and expanded, and compared to that underlying the water flow in order to better understand the pneumatic processes within a stem sample. Moreover, a new and simple equation for gas flow in vessels was derived based on the molar gas flow (mol s-1) rather than volume flow, because the former remains constant with distance throughout the stem axis. We strongly recommend using the Pneumatron in future studies owing to its low cost, convenience, rapidity, and simple operation. However, a number of potential issues need to be considered to avoid artifacts during measurements.

Bamboos are arborescent monocotyledons that have no secondary growth, but can continually produce conduits with diameters appropriate to the current size of the plant. Here, we studied bamboo hydraulic architecture to address the mechanisms involved in compensating for the increase in hydraulic resistance during ontogeny. We measured the hydraulic weighted vessel diameters (Dh ) at different distances from the apex along the stem of Bambusa textilis. The hydraulic resistance of different components and individuals of different heights were quantified using the high-pressure flowmeter method. The Dh showed tip-to-base widening with a scaling exponent in the range of those reported for trees. Although theoretical hydraulic conductivity decreased from base-to-tip, leaf-specific conductivity did not change. Leaves contributed the most to the whole-shoot hydraulic resistance, followed by the leaf-bearing branches. Roots contributed c. 13% to whole-plant resistance. Interestingly, taller individuals showed lower whole-shoot resistance owing to an increased number of resistances in parallel (side-branches), while leaf-specific resistance was independent of plant size. Tip-to-base vessel widening and height-independent constant leaf-specific conductance could be mechanisms for hydraulic optimization in B. textilis. Similar patterns have also been found in woody plants with secondary growth, but this bamboo exhibits them without secondary growth.

Knowledge about the length of xylem vessels is essential to understand water transport in plants because these multicellular units show a 100-fold variation, from less than a centimeter to many meters. However, the available methods to estimate vessel length (VL) distribution are excessively time consuming and do not allow large and in-depth surveys. Here, we describe a semi-automated method to measure hydraulically-weighted VL (VLH) using an automated Pneumatron device. Gas conductivity of a xylem tissue with a certain length is estimated with the Pneumatron in a straightforward and precise way, theoretically similar to the air-injection method. Besides giving results similar to the silicone-injection method, the pneumatic-based method enables faster and easier measurements of more than 50 samples per day, which is a significant advantage. Herein, a detailed description of the methodology is presented as well as the software and an R-script for data analysis. The method described represents an important contribution to studies on plant hydraulic architecture and can improve our understanding about the role of VLH in plant performance under varying water availability.

The pneumatic method has been introduced to quantify embolism resistance in plant xylem of various organs by applying a partial vacuum to cut-open xylem. Despite the similarity in vulnerability curves between the pneumatic and other methods, a modeling approach is needed to investigate if changes in xylem embolism during dehydration can be accurately quantified based on gas diffusion kinetics. Therefore, a unit pipe pneumatic (UPPn) model was developed to estimate gas extraction from intact conduits, which were axially interconnected by inter-conduit pit membranes to cut-open conduits. The physical laws used included Fick’s law for diffusion, Henry’s law for gas concentration partitioning between liquid and gas phases at equilibrium and the ideal gas law. The UPPn model showed that 91% of the extracted gas came from the first five series of embolized, intact conduits and only 9% from the aqueous phase after 15 s of simulation. Considering alternative gas sources, embolism resistance measured with a pneumatron device was systematically overestimated by 2–17%, which corresponded to a typical measuring error of 0.11 MPa for P50 (the water potential equivalent to 50% of the maximum amount of gas extracted). It is concluded that pneumatic vulnerability curves directly measure embolism of intact conduits due to the fast movement of gas across interconduit pit membranes, while gas extraction from sap and diffusion across hydrated cell walls is about 100 times slower. We expect that the UPPn model will also contribute to the understanding of embolism propagation based on temporal gas dynamics.

A long-established ecological paradigm predicts a functional relationship determining vulnerability to cavitation: vulnerability increases with vessel hydraulic efficiency and vessel diameter. Even within a species, big vessels cavitate before small ones.Some centrifuge methods for measuring vulnerability are prone to artifacts due to nano-particles seeding early embolism, as the particles are drawn into vessels during measurements. Both the Sperry and Cochard rotors are prone to early cavitation due to nano-particles drawn into long and wide vessels in Robinia pseudoacacia and Quercus acutissima, whereas extraction centrifuge methods produce vulnerability curves more resistant to cavitation.Sufficient nano-particles pass through the stems to seed early embolism in all rotor designs. For several years, people have thought that early embolism is induced by nano-particles present in laboratory water. One new hypothesis is that the origin of nano-particles is from cut-open living cells but a much bigger study including many species is required to confirm this idea. This paper confirms the hypothesis in comparisons between short-vesselled Acer, and long-vesselled Robinia, and Quercus. Our new results and a review of old results justifies bigger study.Hypothetical nano-particles might explain why different methods for measuring vulnerability curves cause different T50 = tensions causing 50% loss of hydraulic conductivity. Hence the hypothesis for future research should be that the open-vessel artifact is consistent with ‘long’ vessels surrounded by cut open living cells.One sentence SummaryNano-particles induced early cavitation in species with vessel lengths about ¼ the stem length used in all centrifuge rotors, and the origin of nano-particles might be from living cells nearby vessels

We derived a steady-state model of whole root pressure generation through the combined action of all parallel segments of fine roots. This may be the first complete analytical solution for root pressure, which can be applied to complex roots/shoots.The osmotic volume of a single root is equal to that of the vessel lumen in fine roots and adjacent apoplastic spaces. Water uptake occurs via passive osmosis and active solute uptake (, osmol s−1), resulting in the osmolal concentration Cr (mol·kg−1 of water) at a fixed osmotic volume. Solute loss occurs via two passive processes: radial diffusion of solute Km(Cr-Csoil), where Km is the diffusional constant and Csoil is the soil-solute concentration) from fine roots to soil and mass flow of solute and water into the whole plant from the end of the fine roots.The proposed model predicts the quadratic function of root pressure , where b and c are the functions of plant hydraulic resistance, soil water potential, solute flux, and gravitational potential.The present study investigates the theoretical dependencies of Pr on the factors detailed above and demonstrates the root pressure-mediated distribution of water through the hydraulic architecture of a 6.8-m-tall bamboo shoot.One sentence SummaryAn analytical solution for root pressure is derived and illustrated by applying it to the measured hydraulic architecture of a 7-meter bamboo shoot.

Lignified cell walls appear to be impermeable to solutes with molecular weight >300, so ray cell to xylem vessel transport is restricted to unlignified pit membranes., The functional role of pits between living and dead cells has been inferred from anatomical studies but amassing physiological evidence has been challenging. Centrifugation methods were used to strip water from xylem conduits, permitting a more quantitative gravimetric determination of the water and solid contents of cell walls than is possible by more traditional methods. Quantitative anatomical evidence was used to evaluate the water volume in xylem conduits and the water content of living cells. Quantitative perfusion of stems with polyethylene glycol of different molecular weight was used to determine the solute-free space. We measured the portioning of water and solute-free space among anatomical components in stems and demonstrated that lignin impeded the free movement of solutes with molecular weight >300. Hence, movement of large solutes from living cells to xylem conduits is necessarily confined to pit structures that permit transmembrane solute transport via primary walls without lignin. The functional role of pits was additionally indicated by combining data in this paper with previous studies of unusual osmotic relationships in woody species that lack pits between dead wood fibers and vessels. The absence of pits, combined with the evidence of exclusion of solutes of molecular weight >300, explains the unexpected osmotic properties.

Xylem vulnerability to embolism represents an important trait to determine species distribution patterns and drought resistance. However, estimating embolism resistance frequently requires time-consuming and ambiguous hydraulic lab measurements. Based on a recently developed pneumatic method, we present and test the "Pneumatron", a device that generates high time-resolution and fully automated vulnerability curves. Embolism resistance is estimated by applying a partial vacuum to extract air from an excised xylem sample, while monitoring the pressure change over time. Although the amount of gas extracted is strongly correlated with the percentage loss of xylem conductivity, validation of the Pneumatron was performed by comparison with the optical method for Eucalyptus camaldulensis leaves. The Pneumatron improved the precision of the pneumatic method considerably, facilitating the detection of small differences in the (percentage of air discharged [PAD] < 0.47%). Hence, the Pneumatron can directly measure the 50% PAD without any fitting of vulnerability curves. PAD and embolism frequency based on the optical method were strongly correlated (r2 = 0.93) for E. camaldulensis. By providing an open source platform, the Pneumatron represents an easy, low-cost, and powerful tool for field measurements, which can significantly improve our understanding of plant-water relations and the mechanisms behind embolism.

Significant improvements to the centrifuge water-extraction method of measuring the percentage loss volume of water (PLV) and corresponding vulnerability curves (VCs) are reported. Cochard and Sperry rotors are both incapable of measuring the VCs of species with long vessels because of premature embolism induced by hypothetical nanoparticles that can be drawn into segments during flow measurement. In contrast, water extraction pushes nanoparticles out of the sample. This study focuses on a long-vessel species, Robinia pseudoacacia, for which many VCs have been constructed by different methods, and the daily water relations have been quantified. PLV extraction curves have dual Weibull curves, and this paper focuses on the second Weibull curve because it involves the extraction of water from vessels, as proven by staining methods. We demonstrate an improved water extraction method after evaporation correction that has accuracy to within 0.5%, shows good agreement with two traditional methods that are slower and less accurate, and is immune to nanoparticle artefacts. Using Poiseuille's Law and the geometry of vessels, we argue that the percentage loss of conductivity (PLC) equals 2PLV-PLV2 in a special case where all vessels, regardless of size, have the same vulnerability curve. In this special case, this equation predicts the data reasonably well.

Hydraulic safety and efficiency have become the central concept of the interpretation of the structure and function of vessels and their interconnections. Plants form an appropriate xylem network structure to maintain a balance of hydraulic safety vs efficiency. The term ‘tracheid bridge’ is used to describe a possible pathway of water transport between neighboring vessels via tracheids, and this pathway could also provide increased safety against embolisms. However, the only physiological study of such a structure thus far has been in Hippophae rhamnoides Linn. To test the function of tracheid bridges, this research examined four species that have relatively long and solitary vessels, which are two of the criteria for efficient tracheid bridges. Tracheids contributed less than 2.2% of the total conductance of the vessels in these species, but in theory, tracheids could serve as very efficient transport connector pathways that may or may not make direct vessel-to-vessel contact via pit fields between adjacent vessels. In some species, tracheid bridges may represent the dominant pathway for water flow between vessels, whereas in other species, tracheid bridges may be sub-dominant or virtually nil. Broader searches of woody taxa are needed to reveal the functional importance of tracheid bridges in vascular plants.

After drought-induced embolism and repair, tree xylem may be weakened against future drought events (cavitation fatigue). As there are few data on cavitation fatigue in conifers available, we quantified vulnerability curves (VCs) after embolism/repair cycles on eight European conifer species. We induced 50% and 100% loss of conductivity (LC) with a cavitron, and analyzed VCs. Embolism repair was obtained by vacuum infiltration. All species demonstrated complete embolism repair and a lack of any cavitation fatigue after 50% LC . After 100% LC, European larch (Larix decidua), stone pine (Pinus cembra), Norway spruce (Picea abies), and silver fir (Abies alba) remained unaffected, while mountain pine (Pinus mugo), yew (Taxus baccata), and common juniper (Juniperus communis) exhibited 0.4–0.9 MPa higher vulnerability to embolism. A small cavitation fatigue observed in Scots pine (Pinus sylvestris) was probably biased by incomplete embolism repair, as indicated by a correlation of vulnerability shifts and conductivity restoration. Our data demonstrate that cavitation fatigue in conifers is species-specific and depends on the intensity of preceding LC. The lack of fatigue effects after moderate LC, and relevant effects in only three species after high LC, indicate that conifers are relatively resistant against cavitation fatigue. This is remarkable considering the complex and delicate conifer pit architecture and may be important considering climate change projections.

Clusia uvitana Pittíer (Clusiacea) is a tropical hemiepiphyte that has been shown to display a high plasticity in the expression of CAM in response to the environment. When water is available CO

In semi-arid ecoregions, trees are restricted to river valley floodplains where river water supplements the limited precipitation. To characterize the associated diurnal and seasonal dynamics in hydrology and water relations, we studied narrowleaf cottonwoods (Populus angustifolia) along a prairie river in Canada. From June through August, the shallow soil moisture was depleted but moisture remained higher above the alluvial groundwater table, which dropped to 2.3 m along with river recession. Throughout the summer, with the daily rise in temperature and insolation, foliar stomatal conductance (gs) and transpiration (E) increased to midday and then fell, thus maintaining the midday leaf water potential (Ψmd) above ~−1.7 MPa. This Ψmd approximated the water potential associated with 12% loss of xylem conductivity due to cavitation for branches (P12); the Ψmd and P12 varied independently across eight trees, providing differences in relative hydraulic risk. Sap flux density (Fd) was measured with thermal dissipation probes, and revealed diurnal patterns similar to foliar E. In contrast to our expectation, the daily Fd maxima were consistent through the summer despite the seasonal recession in water supply. Canopy conductances (GS), derived from Fd, sapwood area and canopy area, declined with vapor pressure deficit (D) and fell slightly in late summer along with stomatal sensitivity, which reflects the magnitude of decrease in GS with increasing D. For spatial up-scaling, satellite-derived near-infrared reflectance of vegetation revealed the woodland phenology, with leaf expansion from May through June and gradual decline in photosynthetic productivity through the summer. Thus, the phreatophytic cottonwoods: (i) sustained substantial water use and productivity through the warm and dry summer, by (ii) accessing shallow soil moisture and then deeper alluvial groundwater, and (iii) providing diurnal stomatal regulation, to (iv) avoid xylem cavitation and (v) maintain fairly constant hydraulic conductance.

The Pneumatic method has been introduced to quantify embolism resistance in plant xylem of various organs. Despite striking similarity in vulnerability curves between the Pneumatic and hydraulic methods, a modeling approach is highly needed to demonstrate that xylem embolism resistance can be accurately quantified based on gas diffusion kinetics.A Unit Pipe Pneumatic (UPPn) model was developed to estimate gas diffusion from intact conduits, which were axially interconnected by interconduit pit membranes. The physical laws used included Fick’s law for diffusion, Henry’s law for gas concentration partitioning between liquid and gas phases at equilibrium, and the ideal gas law.The UPPn model showed that 91% of the extracted gas came from the first two series of embolized, intact conduits, and only 9% from the aqueous phase after 15 s of simulation. Embolism resistance measured with a Pneumatic apparatus was systematically overestimated by 2 to 17%, corresponding to a typical measuring error of 0.11 MPa for P50 (the water potential equivalent to 50% of the maximum amount of gas extracted).Because results from the UPPn model are supported by experimental evidence, there is a good theoretical and experimental basis for applying the pneumatic method to research on embolism resistance of angiosperms.

Knowledge about the length of xylem vessels is essential to understand water transport in plants because these multicellular units show a 100-fold variation, from less than a centimeter to many meters. However, the available methods to estimate vessel length distribution (VLD) are excessively time consuming and do not allow large and in-depth surveys. Here, we describe a semi-automated method to measure VLD using an automated Pneumatron device. Gas conductivity of a xylem tissue with a certain length is estimated in a straightforward and precise way with the Pneumatron in a way theoretically similar to the air-injection method. The method presented enables fast and easy measurements using multiple devices simultaneously (>50 samples day-1), which is a significant advantage. Here, the apparatus is described in detail as well as how measurements are taken. We also present the software and an R-script for data analysis. The method described represents an important contribution to studies on plant hydraulic architecture and can improve our understanding about the role of VLD in plant performance under varying water availability.

Cavitation resistance is a key trait for characterizing the drought adaption in plants and is usually presented in terms of vulnerability curves. Three principal techniques have been developed to produce vulnerability curves, but curves generated with centrifugation are reported to suffer from artifacts when applied to long-vesseled species. The main cause of this artifact is the issue of open vessels, resulting in a nano-particle effect that may seed premature embolism. We used two methods to test the potential mechanism behind the nano-particle effect in centrifuge-based vulnerability curves. A four-cuvette rotor system based on a traditional Cochard rotor was designed to inhibit effervescence while injecting water, but the recalcitrant vulnerability curves in Robinia could not be eliminated. There may be multiple sources, besides effervescence, of hypothetical nano-particles: they may arise from cut surfaces or they may be always present in the injected water, leading to the premature embolisms. To prevent the entry of the hypothetical nano-particles, water extraction curves in terms of PLV (percentage loss volume of extracted water from stems) vs tensions were constructed. The PLV curves of Robinia showed s-shaped characteristics after subtracting the first Weibull components from water extraction curves, which were not related to the water loss from vessels according to dye staining experiments. The differences between T50 (xylem tension at which 50% of hydraulic conductivity is lost) in mean PLV curve and T50 in percentage loss of conductivity curves determined by the four-cuvette rotor system and by the bench dehydration method were 3.9 MPa and 0.7 MPa, respectively. Hence, PLV curves may be a valid way to measure the cavitation resistance in long-vesseled species with centrifugation. Keeping bark intact in the process of measurement is recommended, otherwise it would increase evaporation from the entire system.

Although cavitation is common in plants, it is unknown whether the cavitation resistance of xylem is seasonally constant or variable. We tested the changes in cavitation resistance of Acer mono before and after a controlled cavitation-refilling and freeze-thaw cycles for a whole year. Cavitation resistance was determined from 'vulnerability curves' showing the percent loss of conductivity versus xylem tension. Cavitation fatigue was defined as a reduction of cavitation resistance following a cavitation-refilling cycle, whereas frost fatigue was caused by a freeze-thaw cycle. A. mono developed seasonal changes in native embolisms; values were relatively high during winter but relatively low and constant throughout the growing season. Cavitation fatigue occurred and changed seasonally during the 12-month cycle; the greatest fatigue response occurred during summer and the weakest during winter, and the transitions occurred during spring and autumn. A. mono was highly resistant to frost damage during the relatively mild winter months; however, a quite different situation occurred during the growing season, as the seasonal trend of frost fatigue was strikingly similar to that of cavitation fatigue. Seasonality changes in cavitation resistance may be caused by seasonal changes in the mechanical properties of the pit membranes.

This paper provides a mini-review of evidence for negative turgor pressure in leaf cells starting with experimental evidence in the late 1950s and ending with biomechanical models published in 2014. In the present study, biomechanical models were used to predict how negative turgor pressure might be manifested in dead tissue, and experiments were conducted to test the predictions. The main findings were as follows: (i) Tissues killed by heating to 60 or 80 °C or by freezing in liquid nitrogen all became equally leaky to cell sap solutes and all seemed to pass freely through the cell walls. (ii) Once cell sap solutes could freely pass the cell walls, the shape of pressure-volume curves was dramatically altered between living and dead cells. (iii) Pressure-volume curves of dead tissue seem to measure negative turgor defined as negative when inside minus outside pressure is negative. (iv) Robinia pseudoacacia leaves with small palisade cells had more negative turgor than Metasequoia glyptostroboides with large cells. (v) The absolute difference in negative turgor between R. pseudoacacia and M. glyptostroboides approached as much as 1.0 MPa in some cases. The differences in the manifestation of negative turgor in living versus dead tissue are discussed.

Pressure-volume (PV) curve analysis is the most common and accurate way of estimating all components of the water relationships in leaves (water potential isotherms) as summarized in the Höfler diagram. PV curve analysis yields values of osmotic pressure, turgor pressure, and elastic modulus of cell walls as a function of relative water content. It allows the computation of symplasmic/apoplastic water content partitioning. For about 20 years, cavitation in xylem has been postulated as a possible source of error when estimating the above parameters, but, to the best of the authors' knowledge, no one has ever previously quantified its influence. Results in this paper provide independent estimates of osmotic pressure by PV curve analysis and by thermocouple psychrometer measurement. An anatomical evaluation was also used for the first time to compare apoplastic water fraction estimates from PV analysis with anatomical values. Conclusions include: (i) PV curve values of osmotic pressure are underestimated prior to correcting osmotic pressure for water loss by cavitation in Metasequoia glyptostroboides; (ii) psychrometer estimates of osmotic pressure obtained in tissues killed by freezing or heating agreed with PV values before correction for apoplastic water dilution; (iii) after correction for dilution effects, a solute concentration enhancement (0.27MPa or 0.11 osmolal) was revealed. The possible sources of solute enhancement were starch hydrolysis and release of ions from the Donnan free space of needle cell walls.

Vessel lengths are important to plant hydraulic studies, but are not often reported because of the time required to obtain measurements. This paper compares the fast dynamic method (air injection method) with the slower but traditional static method (rubber injection method). Our hypothesis was that the dynamic method should yield a larger mean vessel length than the static method. Vessel length was measured by both methods in current year stems of Acer, Populus, Vitis and Quercus representing short- to long-vessel species. The hypothesis was verified. The reason for the consistently larger values of vessel length is because the dynamic method measures air flow rates in cut open vessels. The Hagen-Poiseuille law predicts that the air flow rate should depend on the product of number of cut open vessels times the fourth power of vessel diameter. An argument is advanced that the dynamic method is more appropriate because it measures the length of the vessels that contribute most to hydraulic flow. If all vessels had the same vessel length distribution regardless of diameter, then both methods should yield the same average length. This supports the hypothesis that large-diameter vessels might be longer than short-diameter vessels in most species.

Novel features related to turgor loss adjustment and higher ability to modulate leaf-construction costs could improve A. altissima fitness in its invaded range. Phenotypic plasticity has been recently proposed to play an important role in invasion process. More precisely, phenotypic plasticity of alien invasive species could be higher in their exotic rather than native habitat, probably because of the release from biotic or abiotic constraints. In this paper, several plant functional traits were analysed on Ailanthus altissima (Mill.) Swingle, a highly invasive species in Europe, aiming at providing a comparison among key functional traits measured on individuals growing in their native habitat and to test if values of functional traits and their related plasticity are higher in exotic habitats. Our analysis pinpointed that variability of functional traits in the native habitat was mainly driven by different irradiance, temperature and evaporative demand of the studied sites, in accordance to the traits trade-offs in the Leaf Economic Spectrum. Physiological traits related to drought tolerance were different between native and exotic habitats. In the native one, A. altissima relied on osmoregulation processes to adjust leaf turgor loss point (Ψtlp), as the osmotic potential at full turgor (π0) were lower in drier sites. In the exotic habitat, individuals in drier sites had similar π0 but lower wall elasticity (e), suggesting that leaves had thinner cell walls. Moreover, plasticity in specific leaf area and e were higher in the exotic habitat. The novel features related to Ψtlp adjustment and the higher plasticity in traits related to leaf-construction costs could increase A. altissima fitness in response to different environmental conditions in its invasive range.

Although cavitation is common in plants, it is unknown whether the cavitation resistance of xylem is seasonally constant or variable. We tested the changes in cavitation resistance of Acer mono before and after a controlled cavitation-refilling and freeze-thaw cycles for a whole year. Cavitation resistance was determined from 'vulnerability curves' showing the percent loss of conductivity versus xylem tension. Cavitation fatigue was defined as a reduction of cavitation resistance following a cavitation-refilling cycle, whereas frost fatigue was caused by a freeze-thaw cycle. A. mono developed seasonal changes in native embolisms; values were relatively high during winter but relatively low and constant throughout the growing season. Cavitation fatigue occurred and changed seasonally during the 12-month cycle; the greatest fatigue response occurred during summer and the weakest during winter, and the transitions occurred during spring and autumn. A. mono was highly resistant to frost damage during the relatively mild winter months; however, a quite different situation occurred during the growing season, as the seasonal trend of frost fatigue was strikingly similar to that of cavitation fatigue. Seasonality changes in cavitation resistance may be caused by seasonal changes in the mechanical properties of the pit membranes.

Pre-dawn xylem pressures were measured with bubble manometers attached near the stem bases of 32 species of vines on Barro Colorado Island, Panama, to determine if pressures were sufficient to allow for possible refilling of embolized vessels. Of 29 dicotyledonous species 26 exhibited only negative xylem pressures, even pre-dawn during the wet season. In contrast, three members of the Dilleniaceae exhibited positive pre-dawn xylem pressures, with a maximum of 64 kPa in Doliocarpusmajor. A pressure of 64 kPa is sufficient to push water to a height of 6.4 m against gravity, but the specimens reached heights of 18 m. Thus, in all 29 dicotyledons examined, the xylem pressures were not sufficient to refill embolized vessels in the upper stems. In contrast, two of the smaller, non-dicotyledonous vines, the climbing fern Lygodiumvenustrum and the viny bamboo Rhipidocladumracemiflorum, had xylem pressures sufficient to push water to the apex of the plants. Therefore, a root pressure mechanism to reverse embolisms in stem xylem could apply to some but not to most of the climbing plants that were studied.

One role of stems is that of water storage. The water content of stems increases and decreases as xylem water potential increases and decreases, respectively. Hence, a nondestructive method to measure stem water content (StWC) = (volume of water) : (volume of stem), could be useful in monitoring the drought stress status of plants. We introduce a frequency domain inner fringing capacitor-sensor for measuring StWC which operates at 100 MHz frequency. The capacitor-sensor consists of two wave guides (5-mm-wide braided metal) that snugly fit around the surface of a stem with a spacing of 4-5 mm between guides. Laboratory measurements on analog stems reveals that the DC signal output responds linearly to the relative dielectric constant of the analog stem, is most sensitive to water content between the waveguides to a depth of c. 3 mm from the stem surface, and calibrations based on the gravimetric water loss of excised stems of plants revealed a resolution in StWC of < ± 0.001 v/ v. The sensor performed very well on whole plants with a 100-fold increased resolution compared with previous frequency domain and time domain reflectometry methods and, hence, may be very useful for future research requiring nondestructive measurements of whole plants.

The Cohen method of measuring vessel-length distributions is much more accurate than the DD algorithm on integer values, which should be abandoned. More research is needed to get the real distribution of vessel length. Scientists have been measuring the vessel length of plants for more than 50 years. The method involves infusing stem or segments with a visible substance that completely fills vessels cut open at the infusion surface. The number of infused vessels is then quantified versus distance from the infusion surface. A theoretical model is then used to convert the counts of infused vessels to a vessel length distribution. Over the years the methods and theory have changed greatly. The purpose of this review is to give the reader an understanding of why vessel length is important and to provide a theoretical basis for selection of the best method and theory to arrive at vessel length data.

Use of a staining technique confirms an improved water extraction method with evaporation correction that is consistent with vulnerability curves based on branch hydraulic measurements obtained by the bench-top dehydration method., Significant improvements to the centrifuge water-extraction method of measuring the percentage loss volume of water (PLV) and corresponding vulnerability curves (VCs) are reported. Cochard and Sperry rotors are both incapable of measuring the VCs of species with long vessels because of premature embolism induced by hypothetical nanoparticles that can be drawn into segments during flow measurement. In contrast, water extraction pushes nanoparticles out of the sample. This study focuses on a long-vessel species, Robinia pseudoacacia, for which many VCs have been constructed by different methods, and the daily water relations have been quantified. PLV extraction curves have dual Weibull curves, and this paper focuses on the second Weibull curve because it involves the extraction of water from vessels, as proven by staining methods. We demonstrate an improved water extraction method after evaporation correction that has accuracy to within 0.5%, shows good agreement with two traditional methods that are slower and less accurate, and is immune to nanoparticle artefacts. Using Poiseuille’s Law and the geometry of vessels, we argue that the percentage loss of conductivity (PLC) equals 2PLV–PLV2 in a special case where all vessels, regardless of size, have the same vulnerability curve. In this special case, this equation predicts the data reasonably well.

Frost has been shown to cause frost fatigue (reduced cavitation resistance) in branch segments in the lab. Here, we studied the change in cavitation resistance and percent loss of conductivity (PLC) from fall to spring over 2 consecutive years in three diffuse-porous species in situ. We used the cavitron technique to measure P25, P50 and P90 (the xylem pressure causing a 25, 50 and 90% conductivity loss) and PLC and stained functioning vessels. Cavitation resistance was reduced by 64–87% (in terms of P50), depending on the species and year. P25 was impacted the most and P90 the least, changing the vulnerability curves from s- to r-shaped over the winter in all three species. The branches suffered an almost complete loss of conductivity, but frost fatigue did not necessarily occur concurrently with increases in PLC. In two species, there was a trade-off between conduit size and vulnerability. Spring recovery occurred by growth of new vessels, and in two species by partial refilling of embolized conduits. Although newly grown and functioning conduits appeared more vulnerable to cavitation than year-old vessels, cavitation resistance generally improved in spring, suggesting other mechanisms for partial frost fatigue repair.

Wang, M., Tyree, M. T. and Wasylishen, R. E. 2013. Magnetic resonance imaging of water ascent in embolized xylem vessels of grapevine stem segments. Can. J. Plant Sci. 93: 879–893. Temporal and spatial information about water refilling of embolized xylem vessels and the rate of water ascent in these vessels is critical for understanding embolism repair in intact living vascular plants. High-resolution 1H magnetic resonance imaging (MRI) experiments have been performed on embolized grapevine stem segments while they were subjected to refilling at two different applied water pressures in order to investigate these important aspects of embolism repair. Magnetic resonance imaging difference images show that vessels located near the bark tend to refill faster than do inner ones, suggesting that vessel position within the cross section of the stem may affect the refilling process within the vessel. An MRI method for determining the water ascent velocity in each individual embolized xylem vessel is presented. At ambient pressure, the water ascent velocity ranges from 0.0090 to 0.60 mm min−1, but increases to a range of 0.016 to 0.70 mm min−1 at 9.8 kPa above ambient pressure. A steady-state bubble model that offers analytical solutions of the water ascent velocity in embolized xylem vessels is presented; model calculations show that if other parameters are held constant, water ascent velocity is influenced by vessel diameter and position.

Vulnerability curves (VCs) generally can be fitted to the Weibull equation; however, a growing number of VCs appear to be recalcitrant, that is, deviate from a Weibull but seem to fit dual Weibull curves. We hypothesize that dual Weibull curves in Hippophae rhamnoides L. are due to different vessel diameter classes, inter-vessel hydraulic connections or vessels versus fibre tracheids. We used dye staining techniques, hydraulic measurements and quantitative anatomy measurements to test these hypotheses. The fibres contribute 1.3% of the total stem conductivity, which eliminates the hypothesis that fibre tracheids account for the second Weibull curve. Nevertheless, the staining pattern of vessels and fibre tracheids suggested that fibres might function as a hydraulic bridge between adjacent vessels. We also argue that fibre bridges are safer than vessel-to-vessel pits and put forward the concept as a new paradigm. Hence, we tentatively propose that the first Weibull curve may be accounted by vessels connected to each other directly by pit fields, while the second Weibull curve is associated with vessels that are connected almost exclusively by fibre bridges. Further research is needed to test the concept of fibre bridge safety in species that have recalcitrant or normal Weibull curves.

Frost damage to the xylem conduits of trees is a phenomenon of eco-physiological importance. It is often documented in terms of the percentage loss of conductivity (PLC), an indicator of air filling of the conduits. However, trees that refill their conduits in spring could be impacted more by damage to the conduits that reduce cavitation resistance, making them more susceptible to future drought events. We investigated whether ice formation, dynamic flexing of frozen branches or freeze-thaw events could reduce the cavitation resistance (cause "frost fatigue") in first-year shoots of apple (Malus domestica) and clonal hybrid cottonwood (Walker). Frost fatigue was measured in terms of P50 (the negative xylem pressure required to cause a 50 % loss of conductivity). All treatment groups showed significant frost fatigue, with the exception of the pre-flushed, constantly frozen poplar branches. The P50 following freeze treatments was approximately 50 % of the pre-freeze values. The effect tended to be greater in freeze-thawed branches. Dynamic bending of the branches had no effect on either PLC or P50. In three out of four cases, there was a significant correlation between P50 and PLC. Frost fatigue occurred in both apple and poplar, two unrelated species with different drought and frost tolerances, suggesting that it may be a widespread phenomenon that could impact the ecophysiology of temperate forests.

Climate models suggest that more frequent drought events of greater severity and length, associated with climate change, can be expected in the coming decades. Although drought-induced tree mortality has been recognized as an important factor modulating forest demography at the global scale, the mechanisms underlying drought-induced tree mortality remain contentious. Above- and below-ground growth, gas exchange, water relations and carbon reserve accumulation dynamics at the organ and whole-plant scale were quantified in Populus tremuloides and P. balsamifera seedlings in response to severe drought. Seedlings were maintained in drought conditions over one growing and one dormant winter season. Our experiment presents a detailed description of the effect of severe drought on growth and physiological variables, leading to seedling mortality after an extended period of drought and dormancy. After re-watering following the dormant period, drought-exposed seedlings did not re-flush, showing that the root system had died off. The results of this study suggest a complex series of physiological feedbacks between the measured variables in both Populus species. Further, they reveal that reduced reserve accumulation in the root system during drought decreases the conversion of starch to soluble sugars in roots, which may contribute to the root death of drought-exposed seedlings during the dormant season by compromising the frost tolerance of the root system.

The physiological advantages of negative turgor pressure, Pt, in leaf cells are water saving and homeostasis of reactants. This paper advances methods for detecting the occurrence of negative Pt in leaves. Biomechanical models of pressure-volume (PV) curves predict that negative Pt does not change the linearity of PV curve plots of inverse balance pressure, PB, versus relative water loss, but it does predict changes in either the y-intercept or the x-intercept of the plots depending on where cell collapse occurs in the PB domain because of negative Pt. PV curve analysis of Robinia leaves revealed a shift in the x-intercept (x-axis is relative water loss) of PV curves, caused by negative Pt of palisade cells. The low x-intercept of the PV curve was explained by the non-collapse of palisade cells in Robinia in the PB domain. Non-collapse means that Pt smoothly falls from positive to negative values with decreasing cell volume without a dramatic change in slope. The magnitude of negative turgor in non-collapsing living cells was as low as −1.3 MPa and the relative volume of the non-collapsing cell equaled 58% of the total leaf cell volume. This study adds to the growing evidence for negative Pt.

Measuring whole-plant transpiration is highly relevant considering the increasing interest in understanding and improving plant water use at the whole-plant level. We present an original software package (Amalthea) and a design to create a system for measuring transpiration using laboratory balances based on the readily available commodity hardware. The system is modular, capable of multiple-balance syn- chronisation and is highly scalable from one to one hundred units. The software runs under GNU/Linux and it requires little computer resources. Reporting of transpiration rates is based on the linear regressions of data from pre-set intervals which yields high resolution and provides some level of noise filtering. Using a scale with 0.01-g precision we have mea- sured mean rates as low as 0.01 mmol s -1 over intervals of 220 s. Its flexibility accommodates varied applications, such as monitoring nighttime transpiration or long-term drought treatments and would be suitable for measuring a range of plants, from small herbs to potted trees.

• Context Physiological ecologists have been fascinated by height- or position-linked differences of leaf morphology within tall trees >25 m, but the exact cause is still debated, i.e., is it due to light or height-induced water stress?

In this article, we measured the relative growth rate (RGR) of leaves of Robinia pseudoacacia seedlings under well-watered and water-stressed conditions (mid-day Ψ(w) = leaf water potential estimated with a pressure bomb of -0.48 and -0.98 MPa, respectively). Pressure-volume (PV) curves were done on growing leaves at 25, 50 and 95% of the mature size (growth stage) in order to compute solute potential (Ψ) and turgor pressure (Ψ(P) ) as a function of Ψ(w) . The PV curves and diurnal measurements of Ψ(w) and RGR allowed us to evaluate the parameters (cell wall extensibility m and growth turgor threshold Y) of the Lockhart equation, RGR = m(Ψ(P)-Y), at each growth stage. Our data showed that m and Y did change with leaf age, but the changes were slow enough to evaluate m and Y on any given day. We believe this is the first study to provide evidence that the Lockhart equation adequately quantifies leaf growth of trees over a range of time domains. The value of m linearly declined and Y linearly increased with growth stage. Also, mild drought stress caused a decline in m and increase in Y relative to controls. Although water stress caused an osmotic adjustment which, in turn, increased Ψ(P) in stressed plants relative to controls, the RGR and final leaf sizes were reduced in water-stressed plants because of the impact of water stress on decreased m and increased Y.

In plant physiology models involving bubble nucleation, expansion or elimination, it is typically assumed that the surface tension of xylem sap is equal to that of pure water, though this has never been tested. In this study we collected xylem sap from branches of the tree species Populus tremuloides, Betula papyrifera and Sorbus aucuparia over 3 months. We measured the instantaneous surface tension and followed changes over a period of 0.5-5 h using the pendant drop technique. In all three species the instantaneous surface tension was equal to or within a few percent of that of pure water. Further, in B. papyrifera and S. aucuparia the change over time following drop establishment, although significant, was very small. In P. tremuloides, however, there was a steep decline in surface tension over time that leveled off towards values 21-27% lower than that of pure water. This indicated the presence of surfactants. The values were lower for thinner distal branch segments than for proximal ones closer to the trunk. In some species it appears valid to assume that the surface tension of xylem sap is equal to that of water. However, in branch segments of P. tremuloides close to the terminal bud and hence potentially in other species as well, it may be necessary to take into account the presence of surfactants that reduce the surface tension over time.

The purpose of this study was to determine how shading affects the hydraulic and wood-anatomical characteristics of four boreal conifers (Pinus banksiana, Pinus contorta, Picea glauca and Picea mariana) that differ in shade tolerance. Plants were grown in an open field and under a deciduous-dominated overstory for 6 years. Sapwood- and leaf-area specific conductivity, vulnerability curves, and anatomical measurements (light and scanning electron microscopy) were made on leading shoots from six to nine trees of each treatment combination. There was no difference in sapwood-area specific conductivity between open-grown and understory conifers, although two of four species had larger tracheid diameters in the open. Shaded conifers appeared to compensate for small diameter tracheids by changes in pit membrane structure. Scanning electron microscopy revealed that understory conifers had thinner margo strands, greater maximum pore size in the margo, and more torus extensions. All of these trends may contribute to inadequate sealing of the torus. This is supported by the fact that all species showed increased vulnerability to cavitation when grown in the understory. Although evaporative demand in an understory environment is low, a rapid change into fully exposed conditions could be detrimental for shaded conifers.

Summary 1. Habitat specialization models predict that adaptations to environmental conditions explain species distributions. In tropical rainforests, the ability of the seedlings to survive during drought has been shown to be a key determinant of species distributions. We hypothesize that differences among species in their tolerance to low tissue water status is the mechanism underlying differences in performance during drought. 2. To test this hypothesis we quantified tolerance to low leaf water status for over 20 species from central Panama in screenhouse experiments using two different experimental approaches. Results from both approaches were highly correlated with each other. 3. We found that tolerance to low leaf water status correlated with species drought performance in the field and with their distribution across a gradient of dry season length, with the more desiccationtolerant species having higher survival in dry relative to irrigated conditions, and occurring in drier areas. These results support the hypothesis that, in tropical forests, tolerance to low tissue water status governs seedling performance during drought, as well as being a determinant of species distribution patterns. 4. Lower tolerance to low leaf water status was correlated with greater stem hydraulic conductance. In addition, all species tested, including both desiccation-sensitive and desiccation-resistant species, showed similar losses of xylem conductance, about 80%, when near death. These results suggest that a principal mechanism by which desiccation leads to plant mortality is the loss of xylem conductivity.

Rapid and sensitive detection of stress in trees due to irrigation practices, draught, salinity, pollution, lack of nutrients, or diseases may be useful for research and practical purposes. Tree stress could be monitored by following changes in wood water content via time domain reflectometry (TDR). We have searched for a user-friendly and less expensive tool because, although TDR is perhaps the most suitable method, it is too expensive and complicated for everyday use. The objective of this study was to understand the relations between the electrical conductivity (σ stem ) and water content (θ stem ) in tree stem segments of seven species with TDR probes installed. By leaching stem segments with salt solutions or air we were able to change the salinity and water content independently. We have shown that (i) σ stem is more sensitive to changes in θ stem than to changes in salinity of the sap, and (ii) 30-mm-long rods on the TDR probe can sensitively and accurately measure θ stem . We propose that σ stem changes might be used as a proxy for changes in stem water content or stem water potential. Hence, electrical resistivity measurements may substitute for water content measurements with the following advantages: improved accuracy, higher flexibility in probe construction, application to stem diameters

Dwarfing of fruit trees is often achieved through the use of dwarfing rootstocks. Dwarf trees are characterized by sustained reductions in vegetative growth during the lifetime of the tree. The dwarfing mechanism is not well understood, but it has been hypothesized that hydraulic properties of the rootstock and the graft union are involved. It is hypothesized here that leaf- or stem-specific resistance of at least one hydraulic component of the water transport system would be negatively correlated with rootstock 'vigour', and this could be useful for selection of rootstocks. Hydraulic resistance (R) of fully grown apple trees on a variety of rootstocks of different 'vigours' was measured. Most measurements were with the evaporative flux (EF) method, where water uptake measured with sap flow sensors was related to the pressure gradient from soil (taken as pre-dawn leaf) and midday root (taken as covered root-sucker), stem (from covered leaf), and exposed and shaded leaf water potentials (Psi(l)). R of trees on dwarfing M9 rootstock was compared with that of more vigorous MM106 and MM111 rootstocks in Israel and Vermont, USA. In Israel, M9 consistently had higher leaf-specific hydraulic resistance (R(l)) in the soil to scion stem pathway, but this difference was only significant for one summer. R was larger in M9 between the root and stem, implicating the graft union as the site of increased resistance. In Vermont, R(l) of 9- and 10-year-old trees on six rootstocks of various vigours was not consistently related to vigour, and stem-specific resistance (R(s)) increased with increasing vigour. High pressure flow meter (HPFM) measurements gave a lower R than the EF method in all but one case, perhaps indicating a significant amount of xylem dysfunction in these trees, and demonstrated the increased resistivity of stem sections that included dwarf graft unions as compared with non-graft stem sections. It is concluded that stem- and leaf-specific R are not consistently positively correlated with dwarfing, although the increased resistivity of the graft union in dwarfing rootstocks may influence the transport of water and other elements across the graft union, and therefore be involved in the dwarfing mechanism.

Rainfall and soil moisture availability vary greatly both spatially and temporally. They are prime factors influencing species distribution patterns, diversity and habitat associations in a range of biomes, and limit primary productivity in many natural ecosystems, as well as in forestry and agricultural systems (Hawkins et al. 2003, Kozlowski & Pallardy 1997, Lieth 1975). Projections of drying trends, and increased frequency and intensity of drought events with climate and land-use changes (Hulme & Viner 1998, IPCC 2001) have fuelled an increased interest in the role of drought in determining the structure and function of natural and managed forest systems (Allen & Breshears 1998, Miles et al. 2004). Such projections accentuate the need to assess, understand and predict plant reactions to drought, as well as soil moisture variation at different scales.

Cavitation of water in xylem vessels followed by embolism formation has been authenticated for more than 40 years. Embolism formation involves the gradual buildup of bubble pressure (air) to atmospheric pressure as demanded by Henry's law of equilibrium between gaseous and liquid phases. However, the tempo of pressure increase has not been quantified. In this report, we show that the rate of pressurization of embolized vessels is controlled by both fast and slow kinetics, where both tempos are controlled by diffusion but over different spatial scales. The fast tempo involves a localized diffusion from endogenous sources: over a distance of about 0.05 mm from water-filled wood to the nearest embolized vessels; this process, in theory, should take2 min. The slow tempo involves diffusion of air from exogenous sources (outside the stem). The latter diffusion process is slower because of the increased distance of diffusion of up to 4 mm. Radial diffusion models and experimental measurements both confirm that the average time constant is17 h, with complete equilibrium requiring 1 to 2 d. The implications of these timescales for the standard methods of measuring percentage loss of hydraulic conductivity are discussed in theory and deserve more research in future.

A simulation model was written to compute the time-kinetics of turgor pressure, P, change in Chara corallina during cell pressure probe experiments. The model allowed for the contribution of a membrane plus zero, one, or two unstirred layers of any desired thickness. The hypothesis that a cell with an unstirred layer is a composite membrane that will follow the same kind of kinetics with or without unstirred layers was tested. Typical 'osmotic pulse' experiments yield biphasic curves with minimum or maximum pressures, P min(max) , at time t min(max) and a solute exponential decay with halftime T 1/2 , These observed data were then used to compute composite membrane properties, namely the parameters L P = the hydraulic conductance, σ= reflection coefficient and P s = solute permeability using theoretical equations. Using the simulation model, it was possible to fit an experimental data set to the same values of P min(max) , t min(max) and T 1/2 incorporating different, likely values of unstirred layer thickness, where each thickness requires a unique set of plasmalemma membrane values of L p , or and P,. We conclude that it is not possible to compute plasmalemma membrane properties from cell pressure probe experiments without independent knowledge of the unstirred layer thickness.

Plant productivity, distribution and diversity in tropical rain forests correlate with water availability. Water availability is determined by rainfall and also by the available water capacity of the soil. However, while rainfall is recognized as important, linkages between plant distribution and differences among soils in available water capacity have not been demonstrated. One reason for this may be that measurements of soil moisture, such as gravimetric water content, may be overly simplistic. To investigate this, we compared two sites in Panama, Allee and Rio Paja, which have similar rainfall but different plant communities. Soil water release curves were obtained from about −0.1 MPa to −9 MPa, permitting us to calculate available water capacity. The Rio Paja site had 17% greater available water capacity (between −0.1 MPa to −3 MPa), whereas the gravimetric water content at Rio Paja was lower by 16% in rainy season and by 41% at the end of the dry season. Hence soil gravimetric water content and soil available water capacity did not correspond. The results suggest that available water capacity may better predict plant distributions. Hence, whenever possible, available water capacity should be determined in addition to gravimetric water content.

Summary • The leaf hydraulic conductance (Kleaf) is a major determinant of plant water transport capacity. Here, we measured Kleaf, and its basis in the resistances of leaf components, for fully illuminated leaves of five tree species that regenerate in deep shade, and five that regenerate in gaps or clearings, in Panamanian lowland tropical rainforest. We also determined coordination with stomatal characters and leaf mass per area. • Kleaf varied 10-fold across species, and was 3-fold higher in sun- than in shade-establishing species. On average, 12% of leaf hydraulic resistance (= 1/Kleaf) was located in the petiole, 25% in the major veins, 25% in the minor veins, and 39% outside the xylem. Sun-establishing species had a higher proportion of leaf resistance in the xylem. Across species, component resistances correlated linearly with total leaf resistance. • Kleaf correlated tightly with indices of stomatal pore area, indicating a coordination of liquid- and vapor-phase conductances shifted relative to that of temperate woody species. • Leaf hydraulic properties are integrally linked in the complex of traits that define differences in water use and carbon economy across habitats and vegetation zones.

The amount and seasonality of rainfall varies strongly in the tropics, and plant species abundance, distribution and diversity are correlated with rainfall. Drought periods leading to plant stress occur not only in dry forests, but also in moist and even wet forests. We quantified experimentally the effect of drought on survival of first year seedlings of 28 co-occurring tropical woody plant species in the understory of a tropical moist forest. The seedlings were transplanted to plots and subjected to a drought and an irrigation treatment for 22 weeks during the dry season. Drought effects on mortality and wilting behavior varied greatly among species, so that relative survival in the dry treatment ranged from 0% to about 100% of that in the irrigated treatment. Drought stress was the main factor in mortality, causing about 90% (median) of the total mortality observed in the dry treatment. In almost half of the species, the difference in survival between treatments was not significant even after 22 weeks, implying that many of the species are well adapted to drought in this forest. Relative drought survival was significantly higher in species associated with dry habitats than in those associated with wet habitats, and in species with higher abundance on the dry side of the Isthmus of Panama, than in those more abundant on the wet side. These data show that differential species survival in response to drought, combined with variation in soil moisture availability, may be important for species distribution at the local and regional scale in many tropical forests.

Summary • Root hydraulic conductance (Kr) was measured for five understory shrub species of the neotropical moist forest to determine the effects of arbuscular mycorrhizas (AM) for both carbon-rich and carbon-limited host plants. • Kr was measured using a high pressure flow meter (HPFM) for potted plants grown in a factorial combination of AM fungi (presence/absence) and light (3.5 and 30% of full sun, low/high). • AM colonization improved Kr for the more shade-tolerant species plants when growing in low light. By contrast, water uptake efficiency of the light-demanding species was significantly decreased by AM fungi in high light. Regardless of AM colonization, light-demanding species had a lower capacity than shade-tolerant species to meet transpirational demands, and they allocated substantially more to fine root production relative to leaf area when colonized. • The differential effects of AM colonization and light on a species’ root hydraulic conductance in relation to phylogeny and light adaptation demonstrate that AM fungi may be critical in determining early plant succession and community composition not only due to effects on nutrient uptake, but on water uptake as well.

Studies of the desiccation tolerance of the seedlings of five tropical trees were made on potted plants growing in a greenhouse. Pots were watered to field capacity and then dehydrated for 3 to 9 weeks to reach various visual wilting stages, from slightly wilted to dead. Saturated root hydraulic conductance was measured with a high-pressure flowmeter, and whole-stem hydraulic conductance was measured by a vacuum chamber method. Leaf punches (5.6-mm diameter) were harvested for measurement of leaf water potential by a thermocouple psychrometer method and for measurement of fresh and dry weight. In a parallel study, the same five species were studied in a field experiment in the understory of a tropical forest, where these species frequently germinate. Control seedlings were maintained in irrigated plots during a dry season, and experimental plants were grown in similar plots with rain exclusion shelters. Every 2 to 4 weeks, the seedlings were scored for wilt state and survivorship. After a 22-week drought, the dry plots were irrigated for several weeks to verify visual symptoms of death. The field trials were used to rank drought performance of species, and the greenhouse desiccation studies were used to determine the conditions of moribund plants. Our conclusion is that the desiccation tolerance of moribund plants correlated with field assessment of drought-performance for the five species (r 2 > 0.94).

When petioles of transpiring leaves are cut in the air, according to the ‘Scholander assumption’, the vessels cut open should fill with air as the water is drained away by continued transpiration. The distribution of air-filled vessels versus distance from the cut surface should match the distribution of lengths of ‘open vessels’, i.e. vessels cut open when the leaf is excised. Three different methods were used to estimate the length distribution of open vessels and compared it to the observed distribution of embolisms by the cryo-scanning electron microscope (SEM) method. In the cryo-SEM method, petioles are frozen in liquid nitrogen soon after the petiole is cut. The petioles are then cut at different distances from the original cut surface while frozen and examined in a cryo-SEM facility, where it is easy to distinguish vessels filled with air from those filled with ice. In petioles of Acer platanoides and Juglans regia , the distribution of embolized vessels agrees with expectations. This is in contrast to a previous study on sunflower where cryo-SEM results did not agree with expectations. The reason for this disagreement requires further study for a full elucidation.