Excising stem samples underwater at native tension does not induce xylem cavitation

Xylem resistance to water stress‐induced cavitation is an important trait that is associated with drought tolerance of plants. The level of xylem cavitation experienced by a plant is often assessed as the percentage loss in conductivity (PLC) at different water potentials. Such measurements are constructed with samples that are excised underwater at native tensions. However, a recent study concluded that cutting conduits under significant tension induced cavitation, even when samples were held underwater during cutting. This resulted in artificially increased PLC because of what we have termed a ‘tension‐cutting artefact’. We tested the hypothesized tension‐cutting artefact on five species by measuring PLC at native tension compared with after xylem tensions had been relaxed. Our results did not support the tension‐cutting artefact hypothesis, as no differences were observed between native and relaxed samples in four of five species. In a fifth species (Laurus nobilis), differences between native and relaxed samples appear to be due to vessel refilling rather than a tension‐cutting effect. We avoided the tension‐cutting artefact by cutting samples to slightly longer than their measurement length and subsequent trimming of at least 0.5 cm of sample ends prior to measurement.     

Refilling of a hydraulically isolated embolized xylem vessel: model calculations

When they are hydraulically isolated, embolized xylem vessels can be refilled, while adjacent vessels remain under tension. This implies that the pressure of water in the refilling vessel must be equal to the bubble gas pressure, which sets physical constraints for recovery. A model of water exudation into the cylindrical vessel and of bubble dissolution based on the assumption of hydraulic isolation is developed. Refilling is made possible by the turgor of the living cells adjacent to the refilling vessel, and by a reflection coefficient below 1 for the exchange of solutes across the interface between the vessel and the adjacent cells. No active transport of solutes is assumed. Living cells are also capable of importing water from the water-conducting vessels. The most limiting factors were found to be the osmotic potential of living cells and the ratio of the volume of the adjacent living cells to that of the embolized vessel. With values for these of 1.5 MPa and 1, respectively, refilling times were in the order of hours for a broad range of possible values of water conductivity coefficients and effective diffusion distances for dissolved air, when the xylem water tension was below 0.6 MPa and constant. Inclusion of the daily pattern for xylem tension improved the simulations. The simulated gas pressure within the refilling vessel was in accordance with recent experimental results. The study shows that the refilling process is physically possible under hydraulic isolation, while water in surrounding vessels is under negative pressure. However, the osmotic potentials in the refilling vessel tend to be large (in the order of 1 MPa). Only if the xylem water tension is, at most, twice atmospheric pressure, the reflection coefficient remains close to 1 (0.95) and the ratio of the volume of the adjacent living cells to that of the embolized vessel is about 2, does the osmotic potential stay below 0.4 MPa.     

In vivo magnetic resonance imaging of xylem vessel contents in woody lianas

Previous reports suggest that in some plant species the refilling of embolized xylem vessels can occur while negative pressure exists in the xylem. The aim of this experiment was to use non‐destructive nuclear magnetic resonance imaging (MRI) to study the dynamics of xylem cavitation and embolism repair in‐vivo. Serial ¹H‐MRI was used to monitor the contents of xylem vessels in stems of two dicotyledonous (Actinidia deliciosa and Actinidia chinensis, kiwifruit) and one monocotyledonous (Ripogonum scandens, supplejack) species of woody liana. The configuration of the horizontal wide bore magnet and probe allowed the imaging of woody stems up to 20 mm in diameter. Tests using excised stems confirmed that the image resolution of 78 µm and digital image subtraction could be used to detect the emptying and refilling of individual vessels. Imaging was conducted on both intact plants and excised shoots connected to a water supply. In the case of Ripogonum the excised shoots were long enough to allow the distal end of the shoot, including all leaves, to be exposed to ambient conditions outside the building while the proximal end was inside the MRI magnet. In total, six stems were monitored for 240 h while the shoots were subjected to treatments that included light and dark periods, water stress followed by re‐watering, and the covering of all leaves to prevent transpiration. The sudden emptying of water‐filled vessels occurred frequently while xylem water potential was low (below ?0.5 MPa for Actinidia, ?1.0 MPa for Ripogonum), and less frequently after xylem water potential approached zero at the end of water‐stress treatments. No refilling of empty vessels was observed at any time in any of the species examined. It is concluded that embolism repair under negative pressure does not occur in the species examined here. Embolism repair may be more likely in species with narrower xylem vessels, but further experiments are required with other species before it can be concluded that repair during transpiration is a widespread phenomenon.     

Relax and refill: xylem rehydration prior to hydraulic measurements favours embolism repair in stems and generates artificially low PLC values

Diurnal changes in percentage loss of hydraulic conductivity (PLC), with recorded values being higher at midday than on the following morning, have been interpreted as evidence for the occurrence of cycles of xylem conduits' embolism and repair. Recent reports have suggested that diurnal PLC changes might arise as a consequence of an experimental artefact, that is, air entry into xylem conduits upon cutting stems, even if under water, while under substantial tension generated by transpiration. Rehydration procedures prior to hydraulic measurements have been recommended to avoid this artefact. In the present study, we show that xylem rehydration prior to hydraulic measurements might favour xylem refilling and embolism repair, thus leading to PLC values erroneously lower than those actually experienced by transpiring plants. When xylem tension relaxation procedures were performed on stems where refilling mechanisms had been previously inhibited by mechanical (girdling) or chemical (orthovanadate) treatment, PLC values measured in stems cut under native tension were the same as those measured after sample rehydration/relaxation. Our data call for renewed attention to the procedures of sample collection in the field and transport to the laboratory, and suggest that girdling might be a recommendable treatment prior to sample collection for PLC measurements.     

Refilling of Embolized Vessels in Young Stems of Laurel. Do We Need a New Paradigm?

Recovery of hydraulic conductivity after the induction of embolisms was studied in woody stems of laurel (Laurus nobilis). Previous experiments confirming the recovery of hydraulic conductivity when xylem pressure potential was less than −1 MPa were repeated, and new experiments were done to investigate the changes in solute composition in xylem vessels during refilling. Xylem sap collected by perfusion of excised stem segments showed elevated levels of several ions during refilling. Stem segments were frozen in liquid N₂ to view refilling vessels using cryoscanning electron microscopy. Vessels could be found in all three states of presumed refilling: (a) mostly water with a little air, (b) mostly air with a little water, or (c) water droplets extruding from vessel pits adjacent to living cells. Radiographic probe microanalysis of refilling vessels revealed nondetectable levels of dissolved solutes. Results are discussed in terms of proposed mechanisms of refilling in vessels while surrounding vessels were at a xylem pressure potential of less than −1 MPa. We have concluded that none of the existing paradigms explains the results.     

Cutting xylem under tension or supersaturated with gas can generate PLC and the appearance of rapid recovery from embolism

We investigated the common assumption that severing stems and petioles under water preserves the hydraulic continuity in the xylem conduits opened by the cut when the xylem is under tension. In red maple and white ash, higher percent loss of conductivity (PLC) in the afternoon occurred when the measurement segment was excised under water at native xylem tensions, but not when xylem tensions were relaxed prior to sample excision. Bench drying vulnerability curves in which measurement samples were excised at native versus relaxed tensions showed a dramatic effect of cutting under tension in red maple, a moderate effect in sugar maple, and no effect in paper birch. We also found that air injection of cut branches (red and sugar maple) at pressures of 0.1 and 1.0 MPa resulted in PLC greater than predicted from vulnerability curves for samples cut 2 min after depressurization, with PLC returning to expected levels for samples cut after 75 min. These results suggest that sampling methods can generate PLC patterns indicative of repair under tension by inducing a degree of embolism that is itself a function of xylem tensions or supersaturation of dissolved gases (air injection) at the moment of sample excision. Implications for assessing vulnerability to cavitation and levels of embolism under field conditions are discussed.     

In vivo observation of cavitation and embolism repair using magnetic resonance imaging

Magnetic resonance imaging (MRI) was used to noninvasively monitor the status of individual xylem vessels in the stem of an intact, transpiring grape (Vitis vinifera) plant over a period of approximately 40 h. Proton density-weighted MRI was used to visualize the distribution of mobile water in the stem and individual xylem vessels were scored as either water or gas filled (i.e. embolized). The number of water-filled vessels decreased during the first 24 h of the experiment, indicating that approximately 10 vessels had cavitated during this time. Leaf water potentials decreased from -1.25 to -2.1 MPa during the same period. Watering increased leaf water potentials to -0.25 MPa and prevented any further cavitation. Refilling of xylem vessels occurred as soon as the lights were switched off, with the majority of vessels becoming refilled with water during the first 2 to 3 h in darkness. These measurements demonstrate that MRI can be used to monitor the functional status of individual xylem vessels, providing the first method to study the process of cavitation and embolism repair in intact plants.     

In vivo visualization of the water-refilling process in xylem vessels using X-ray micro-imaging

BACKGROUND AND AIMS: Xylem vessels containing gases (embolized) must be refilled with water if they are to resume transport of water through the plant, so refilling is of great importance for the maintenance of water balance in plants. However, the refilling process is poorly understood because of inadequate examination methods. Simultaneous measurements of plant anatomy and vessel refilling are essential to elucidate the mechanisms involved. In the present work, a new technique based on phase-contrast X-ray imaging is presented that visualizes, in vivo and in real time, both xylem anatomy and refilling of embolized vessels. METHODS: With the synchrotron X-ray micro-imaging technique, the refilling of xylem vessels of leaves and a stem of Phyllostachys bambusoides with water is demonstrated under different conditions. The technique employs phase contrast imaging of X-ray beams, which are transformed into visible light and are photographed by a charge coupled device camera. X-ray images were captured consecutively at every 0.5 s with an exposure time of 10 ms. KEY RESULTS: The interface (meniscus) between the water and gas phases in refilling the xylem vessels is displayed. During refilling, the rising menisci in embolized vessels showed repetitive flow, i.e. they temporarily stopped at the end walls of the vessel elements while gas bubbles were removed. The meniscus then passed through the end wall at a faster rate than the speed of flow in the main vessels. In the light, the speed of refilling in a specific vessel was slower than that in the dark, but this rate increased again after repeated periods in darkness. CONCLUSIONS: Real-time, non-destructive X-ray micro-imaging is an important, useful and novel technique to study the relationship between xylem structure and the refilling of embolized vessels in intact plants. It provides new insight into understanding the mechanisms of water transport and the refilling of embolized vessels, which are not understood well.     

The relationship between xylem conduit diameter and cavitation caused by freezing

The centrifuge method for measuring the resistance of xylem to cavitation by water stress was modified to also account for any additional cavitation that might occur from a freeze-thaw cycle. A strong correlation was found between cavitation by freezing and mean conduit diameter. On the one extreme, a tracheid-bearing conifer and diffuse-porous angiosperms with small-diameter vessels (mean diameter <30 μm) showed no freezing-induced cavitation under modest water stress (xylem pressure = −0.5 MPa), whereas species with larger diameter vessels (mean >40 μm) were nearly completely cavitated under the same conditions. Species with intermediate mean diameters (30–40 μm) showed partial cavitation by freezing. These results are consistent with a critical diameter of 44 μm at or above which cavitation would occur by a freeze–thaw cycle at −0.5 MPa. As expected, vulnerability to cavitation by freezing was correlated with the hydraulic conductivity per stem transverse area. The results confirm and extend previous reports that small-diameter conduits are relatively resistant to cavitation by freezing. It appears that the centrifuge method, modified to include freeze–thaw cycles, may be useful in separating the interactive effects of xylem pressure and freezing on cavitation.     

Intra- and inter-plant variation in xylem cavitation in Betula occidentalis

A modified version of a method that uses positive air pressures to determine the complete cavitation response of a single axis is presented. Application of the method to Betula occidentalis Hook, gave a cavitation response indistinguishable from that obtained by dehydration, thus verifying the technique and providing additional evidence that cavitation under tension occurs by air entry through interconduit pits. Incidentally, this also verified pressure-bomb estimates of xylem tension and confirmed the existence of large (i.e. >0·4 MPa) tensions in xylem, which have been questioned in recent pressure-probe studies. The air injection method was used to investigate variation within and amongst individuals of B. occidentalis. Within an individual, the average cavitation tension increased from 0·66±0·27 MPa in roots (3·9 to 10·7 mm diameter), to 1·17±0·10 MPa in trunks (12 to 16 mm diameter), to 1·36±0·04 MPa in twigs (3·9 to 5 mm diameter). Cavitation tension was negatively correlated with the hydraulically weighted mean of the vessel diameter, and was negatively correlated with the conductance of the xylem per xylem area. Native cavitation was within the range predicted from the measured cavitation response and in situ maximum xylem tensions: roots were significantly cavitated compared with minimal cavitation in trunks and twigs. Leaf turgor pressure declined to zero at the xylem tensions predicted to initiate cavitation in petiole xylem (1·5 MPa). Amongst individuals within B. occidentalis, average cavitation tension in the main axis varied from 0·90 to 1·90 MPa and showed no correlation with vessel diameter. The main axes of juveniles (2–3 years old) had significantly narrower vessel diameters than those of adults, but there was no difference in the average cavitation tension. However, juvenile xylem retained hydraulic conductance to a much higher xylem tension (3·25 MPa) than did adult xylem (2·25 MPa), which could facilitate drought survival during establishment.     

Are leaves ‘freewheelin'? Testing for a Wheeler‐type effect in leaf xylem hydraulic decline

A recent study found that cutting shoots under water while xylem was under tension (which has been the standard protocol for the past few decades) could produce artefactual embolisms inside the xylem, overestimating hydraulic vulnerability relative to shoots cut under water after relaxing xylem tension (Wheeler et al. 2013). That study also raised the possibility that such a ‘Wheeler effect’ might occur in studies of leaf hydraulic vulnerability. We tested for such an effect for four species by applying a modified vacuum pump method to leaves with minor veins severed, to construct leaf xylem hydraulic vulnerability curves. We tested for an impact on leaf xylem hydraulic conductance (K_x) of cutting the petiole and minor veins under water for dehydrated leaves with xylem under tension compared with dehydrated leaves after previously relaxing xylem tension. Our results showed no significant ‘cutting artefact’ for leaf xylem. The lack of an effect for leaves could not be explained by narrower or shorter xylem conduits, and may be due to lesser mechanical stress imposed when cutting leaf petioles, and/or to rapid refilling of emboli in petioles. These findings provide the first validation of previous measurements of leaf hydraulic vulnerability against this potential artefact.     

Xylem Hydraulic Characteristics, Water Relations and Wood Anatomy of the Resurrection PlantMyrothamnus flabellifoliusWelw.

Myrothamnus flabellifoliusWelw. is a desiccation-tolerant (‘resurrection’)plant with a woody stem. Xylem vessels are narrow (14 µmmean diameter) and perforation plates are reticulate. This leadsto specific and leaf specific hydraulic conductivities thatare amongst the lowest recorded for angiosperms (k_s0.87 kg m^-1MPa^-1s^-1;k_l3.28x10^-5kg m^-1MPa^-1s^-1, stem diameter 3 mm). Hydraulic conductivitiesdecrease with increasing pressure gradient. Transpiration ratesin well watered plants were moderate to low, generating xylemwater potentials of -1 to -2 MPa. Acoustic emissions indicatedextensive cavitation events that were initiated at xylem waterpotentials of -2 to -3 MPa. The desiccation-tolerant natureof the tissue permits this species to survive this interruptionof the water supply. On rewatering the roots pressures thatwere developed were low (2.4 kPa). However capillary forceswere demonstrated to be adequate to account for the refillingof xylem vessels and re-establishment of hydraulic continuityeven when water was under a tension of -8 kPa. During dehydrationand rehydration cycles stems showed considerable shrinking andswelling. Unusual knob-like structures of unknown chemical compositionwere observed on the outer surface of xylem vessels. These maybe related to the ability of the stem to withstand the mechanicalstresses associated with this shrinkage and swelling.Copyright1998 Annals of Botany Company cavitation, desiccation, hydraulic conductivity, refilling, resurrection plant, root pressure, xylem anatomy,Myrothamnus flabellifolius     

Canny's compensating pressure theory fails a test

Canny's compensating pressure theory for water transport (American Journal of Botany 85: 897–909) has evolved from the premise that cavitation pressures are only a few tenths of a megapascal negative (approximately −0.3 MPa). In contradiction, “vulnerability curves” indicate that xylem pressures can drop below −3 MPa in some species without causing a loss of hydraulic conductivity. Canny claims these curves do not measure the limits to negative pressure by cavitation, but rather the limits to the compensating tissue pressure that otherwise quickly refills cavitated conduits. Compensating pressure is derived from the turgor pressure of the living cells in the tissue. To test this claim, we compared vulnerability curves of Betula nigra stems given three treatments: (1) living control, (2) killed in a microwave oven, and (3) perfused with a −1.5 MPa (10% w/w) mannitol solution. According to Canny's theory, the microwaved and mannitol curves should show cavitation and loss of conductance beginning at approximately −0.3 MPa because in both cases, the turgor pressure would be eliminated or substantially reduced compared to controls. We also tested the refilling capability of nonstressed stems where compensating pressure would be in full operation and compared this with dead stems with no compensating pressure. According to Canny's interpretation of vulnerability curves, the living stems should refill within 5 min. Results failed to support the compensating tissue theory because (a) all vulnerability curves were identical, reaching a −1.5 MPa threshold before substantial loss of conductance occurred, and (b) killed or living stems had equally slow refilling rates showing no significant increase in conductivity after 30 min. In consequence, the cohesion theory remains the most parsimonious explanation of xylem sap ascent in plants.     

Vulnerability to cavitation differs between current‐year and older xylem: non‐destructive observation with a compact magnetic resonance imaging system of two deciduous diffuse‐porous species

Development of xylem embolism during water stress in two diffuse‐porous hardwoods, Katsura (Cercidiphyllum japonicum) and Japanese white birch (Betula platyphylla var. japonica), was observed non‐destructively under a compact magnetic resonance imaging (MRI) system in addition to conventional quantitation of hydraulic vulnerability to cavitation from excised stem segments. Distribution of white and dark areas in MR images corresponded well to the distribution of water‐filled/embolized vessels observed by cryo‐scanning electron microscopy in both species. Water‐filled vessels were observed in MR images as white areas in Katsura and as white dots in Japanese white birch, respectively, and embolisms could be detected as a change to dark areas. The increase in the relative embolized area (REA: %) in the cross‐sectional area of total xylem during water stress, which was estimated from the binarized MR images, was consistent with the hydraulic vulnerability curves of these species. From the non‐destructive MRI observations, cavitation induced by water stress was shown to develop earlier in 1‐ or 2‐year‐old xylem than in the current‐year xylem in both species; that is, the vulnerability to cavitation differs between vessels in the current‐year xylem and those in older annual rings.     

Xylem vulnerability to cavitation can be accurately characterised in species with long vessels using a centrifuge method

Vulnerability to cavitation curves describe the decrease in xylem hydraulic conductivity as xylem pressure declines. Several techniques for constructing vulnerability curves use centrifugal force to induce negative xylem pressure in stem or root segments. Centrifuge vulnerability curves constructed for long‐vesselled species have been hypothesised to overestimate xylem vulnerability to cavitation due to increased vulnerability of vessels cut open at stem ends that extend to the middle or entirely through segments. We tested two key predictions of this hypothesis: (i) centrifugation induces greater embolism than dehydration in long‐vesselled species, and (ii) the proportion of open vessels changes centrifuge vulnerability curves. Centrifuge and dehydration vulnerability curves were compared for a long‐ and short‐vesselled species. The effect of open vessels was tested in four species by comparing centrifuge vulnerability curves for stems of two lengths. Centrifuge and dehydration vulnerability curves agreed well for the long‐ and short‐vesselled species. Centrifuge vulnerability curves constructed using two stem lengths were similar. Also, the distribution of embolism along the length of centrifuged stems matched the theoretical pressure profile induced by centrifugation. We conclude that vulnerability to cavitation can be accurately characterised with vulnerability curves constructed using a centrifuge technique, even in long‐vesselled species.     

The Dynamics of Embolism Repair in Xylem: In Vivo Visualizations Using High-Resolution Computed Tomography

Water moves through plants under tension and in a thermodynamically metastable state, leaving the nonliving vessels that transport this water vulnerable to blockage by gas embolisms. Failure to reestablish flow in embolized vessels can lead to systemic loss of hydraulic conductivity and ultimately death. Most plants have developed a mechanism to restore vessel functionality by refilling embolized vessels, but the details of this process in vessel networks under tension have remained unclear for decades. Here we present, to our knowledge, the first in vivo visualization and quantification of the refilling process for any species using high-resolution x-ray computed tomography. Successful vessel refilling in grapevine (Vitis vinifera) was dependent on water influx from surrounding living tissue at a rate of 6 × 10⁻⁴ μm s⁻¹, with individual droplets expanding over time, filling vessels, and forcing the dissolution of entrapped gas. Both filling and draining processes could be observed in the same vessel, indicating that successful refilling requires hydraulic isolation from tensions that would otherwise prevent embolism repair. Our study demonstrates that despite the presence of tensions in the bulk xylem, plants are able to restore hydraulic conductivity in the xylem.Vascular plants have evolved a simple but elegant system for long-distance transport of water and minerals through a network of nonliving, pipe-like cells. Whereas long-distance transport in animals is actively driven by positive pressure, most water transport in plants is passively driven by tension as explained by the Cohesion-Tension (C-T) theory (Dixon and Joly, 1894; Tyree, 2003). Water under tension is metastable however (Hayward, 1971), making the transport system inherently vulnerable to cavitation and blockage by gas embolisms (Tyree and Sperry, 1989). Direct measurements of negative pressures (tensions) in xylem (Wei et al., 1999) have confirmed the fundamental basis for the C-T theory of water transport in plants (e.g. Tyree, 2003), but many details regarding the susceptibility of the xylem network to cavitation and blockage by embolisms, and a thermodynamically plausible mechanism for the repair of these embolisms, remain unclear (Clearwater and Goldstein, 2005).Plants have apparently evolved mechanisms, including root pressure, to remove embolisms and restore water transport in vessels (Sperry et al., 1987; Tibbetts and Ewers, 2000; Isnard and Silk, 2009). Refilling of embolized vessels far from roots (Holbrook et al., 2001) and under a state of tension (Salleo and Gullo, 1986) is not well understood, but most hypotheses involve localized solute export into embolized vessels from adjacent living xylem parenchyma, osmotic movement of water into these vessels, and isolation of the refilling vessel from the tension in its local water environment (Tyree et al., 1999; Hacke and Sperry, 2003; Clearwater and Goldstein, 2005; Salleo et al., 2006). Embolism repair is complicated by the fact that xylem conduits (tracheids and vessels) form an interconnected network. While such a network will provide a low-resistance pathway for the bulk flow of water when the conduits are filled, if a cavitation event and subsequent embolism (gas bubble) either spontaneously occurs within a conduit, or spreads to it from another conduit, the presence of tension in this network should also quickly drain a conduit of its water and prevent its refilling. The spread of embolisms is limited by the small effective pore size of the connections between conduits (known as pit membranes), but under conditions of low plant water availability, embolisms do occur and spread (Tyree and Zimmermann, 2002; Choat et al., 2008), and evidence for the repair of embolized vessels, despite the presumed presence of a tension throughout the plant xylem, has been obtained in many species (Salleo et al., 1996; McCully et al., 1998; Zwieniecki and Holbrook, 1998; Kaufmann et al., 2009).A major limitation to the testing of these hypotheses and to our understanding of embolism repair has been the lack of in vivo observations at a sufficient resolution and an appropriate temporal scale to document how the refilling occurs. Here we present a new method for imaging the functional status of vessels using high-resolution x-ray computed tomography (HRCT), providing, to our knowledge, the first in vivo visualization of the refilling process for any species. Previous in vivo measurements of vessel refilling have been performed using NMR imaging, but the resolution was insufficient to determine the source of the refilling water (Holbrook et al., 2001; Scheenen et al., 2007). In vivo imaging at this scale allows for nondestructive visualization and measurement of the change in both air and water volume within the vessel lumen, giving unprecedented access to the mechanisms of embolism repair.     

Water transport properties of seven woody species from the semi-arid Mu Us Sandy Land,China

Maintenance of water transport is very important for plant growth and survival. We studied seven woody species that inhabit the semi-arid Mu Us Sandy Land, China, to understand their strategies for maintaining hydraulic function. We evaluated water transport properties, including cavitation resistance, hydraulic recovery, and water loss regulation by stomatal control, which are associated with xylem structural and leaf physiological traits. We also discussed the water-use characteristics of these species by comparing them with those of species in other regions. Species with tracheids had higher levels of xylem resistance to cavitation and a smaller midday transpiration rate than the other species studied. Although species with vessels were less resistant to cavitation, some recovered hydraulic conductivity within 12 h of rehydration. Species with xylem tracheids could maintain their hydraulic function through resistance to cavitation and by relaxing xylem tension. Although species with vessels had less resistant xylem, they could maintain hydraulic function through hydraulic recovery even when xylem dysfunction occurred. Additionally, the species studied here were less resistant to cavitation than species in semi-arid environments, and equally or less resistant than species in humid environments. Rather than allow hydraulic dysfunction due to drought-induced dehydration, they may develop water absorption systems to avoid or recover quickly from hydraulic dysfunction. Thus, not only stem cavitation resistance to drought but also stem–root coordination should be considered when selecting plants for the revegetation of arid regions.     

Relationships among xylem transport, biomechanics and storage in stems and roots of nine Rhamnaceae species of the California chaparral

Here, hypotheses about stem and root xylem structure and function were assessed by analyzing xylem in nine chaparral Rhamnaceae species. Traits characterizing xylem transport efficiency and safety, mechanical strength and storage were analyzed using linear regression, principal components analysis and phylogenetic independent contrasts (PICs). Stems showed a strong, positive correlation between xylem mechanical strength (xylem density and modulus of rupture) and xylem transport safety (resistance to cavitation and estimated vessel implosion resistance), and this was supported by PICs. Like stems, greater root cavitation resistance was correlated with greater vessel implosion resistance; however, unlike stems, root cavitation resistance was not correlated with xylem density and modulus of rupture. Also different from stems, roots displayed a trade-off between xylem transport safety from cavitation and xylem transport efficiency. Both stems and roots showed a trade-off between xylem transport safety and xylem storage of water and nutrients, respectively. Stems and roots differ in xylem structural and functional relationships, associated with differences in their local environment (air vs soil) and their primary functions.     

Rare pits, large vessels and extreme vulnerability to cavitation in a ring-porous tree species

? The rare pit hypothesis predicts that the extensive inter-vessel pitting in large early-wood vessels of ring-porous trees should render many of these vessels extremely vulnerable to cavitation by air-seeding. This prediction was tested in Quercus gambelii. ? Cavitation was assessed from native hydraulic conductivity at field sap tension and in dehydrated branches. Single-vessel air injections gave air-seeding pressures through vessel files; these data were used to estimate air-seeding pressures for inter-vessel walls and pits. ? Extensive cavitation occurred at xylem sap tensions below 1 MPa. Refilling occurred below 0.5 MPa and was inhibited by phloem girdling. Remaining vessels cavitated over a wide range to above 4 MPa. Similarly, 40% of injected vessel files air-seeded below 1.0 MPa, whereas the remainder seeded over a wide range exceeding 5 MPa. Inter-vessel walls averaged 1.02 MPa air-seeding pressure, similar and opposite to the mean cavitation tension of 1.22 MPa. Consistent with the rare pit hypothesis, only 7% of inter-vessel pits were estimated to air-seed by 1.22 MPa. ? The results confirm the rare pit prediction that a significant fraction of large vessels in Q. gambelii experience high probability of failure by air-seeding.     

Cu2+ inhibition of gel secretion in the xylem and its potential implications for water uptake of cut Acacia holosericea stems

Maintaining a high rate of water uptake is crucial for maximum longevity of cut stems. Physiological gel/tylosis formation decreases water transport efficiency in the xylem. The primary mechanism of action for post‐harvest Cu²⁺ treatments in improving cut flower and foliage longevity has been elusive. The effect of Cu²⁺ on wound‐induced xylem vessel occlusion was investigated for Acacia holosericea A. Cunn. ex G. Don. Experiments were conducted using a Cu²⁺ pulse (5 h, 2.2 mM) and a Cu²⁺ vase solution (0.5 mM) vs a deionized water (DIW) control. Development of xylem blockage in the stem‐end region 10 mm proximal to the wounded stem surface was examined over 21 days by light and transmission electron microscopy. Xylem vessels of stems stood into DIW were occluded with gels secreted into vessel lumens via pits from surrounding axial parenchyma cells. Gel secretion was initiated within 1–2 days post‐wounding and gels were detected in the xylem from day 3. In contrast, Cu²⁺ treatments disrupted the surrounding parenchyma cells, thereby inhibiting gel secretion and maintaining the vessel lumens devoid of occlusions. The Cu²⁺ treatments significantly improved water uptake by the cut stems as compared to the control.