Lars H. Wegner

Xylem ionic relations and salinity tolerance in barley

Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non-invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na(+) loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na(+) concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K(+)/Na(+) ratios and efficiently sequester the accumulated Na(+) in leaves. The former is achieved by more efficient loading of K(+) into the xylem. We argue that the observed increases in xylem K(+) and Na(+) concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K(+)-permeable voltage-sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K(+)/Na(+) ratio in the xylem sap.

Functional repair of embolized vessels in maize roots after temporal drought stress, as demonstrated by magnetic resonance imaging

Xylem sap under high tension is in a metastable state and tends to cavitate, frequently leading to an interruption of the continuous water columns. Mechanisms of cavitation repair are controversially discussed. Magnetic resonance (MR) imaging provides a noninvasive, high spatial and temporal resolution approach to monitor xylem cavitation, refilling, and functionality. Spin density maps of drought-stressed maize taproots were recorded to localize cavitation events and to visualize the refilling processes; c. 2 h after release of the nutrient solution from the homemade MR imaging cuvette that received the root, late metaxylem vessels started to cavitate randomly as identified by a loss of signal intensity. After c. 6 h plants were rewatered, leading to a repair of water columns in five out of eight roots. Sap ascent during refilling, monitored with multislice MR imaging sequences, varied between 0.5 mm min(-1) and 3.3 mm min(-1). Flow imaging of apparently refilled vessels was performed to test for functional repair. Occasionally, a collapse of xylem vessels under tension was observed; this collapse was reversible upon rewatering. Refilling was an all-or-none process only observed under low-light conditions. Absence of flow in some of the apparently refilled vessels indicates that functionality was not restored in these particular vessels, despite a recovery of the spin density signal.

Nanosecond electric pulses trigger actin responses in plant cells

We have analyzed the cellular effects of nanosecond pulsed electrical fields on plant cells using fluorescently tagged marker lines in the tobacco cell line BY-2 and confocal laser scanning microscopy. We observe a disintegration of the cytoskeleton in the cell cortex, followed by contraction of actin filaments towards the nucleus, and disintegration of the nuclear envelope. These responses are accompanied by irreversible permeabilization of the plasma membrane manifest as uptake of Trypan Blue. By pretreatment with the actin-stabilizing drug phalloidin, the detachment of transvacuolar actin from the cell periphery can be suppressed, and this treatment can also suppress the irreversible perforation of the plasma membrane. We discuss these findings in terms of a model, where nanosecond pulsed electric fields trigger actin responses that are key events in the plant-specific form of programmed cell death.

Foliar water supply of tall trees: evidence for mucilage-facilitated moisture uptake from the atmosphere and the impact on pressure bomb measurements

Summary.The water supply to leaves of 25 to 60 m tall trees (including high-salinity-tolerant ones) was studied. The filling status of the xylem vessels was determined by xylem sap extraction (using jet-discharge, gravity-discharge, and centrifugation) and by 1H nuclear magnetic resonance imaging of wood pieces. Simultaneously, pressure bomb experiments were performed along the entire trunk of the trees up to a height of 57 m. Clear-cut evidence was found that the balancing pressure (Pb) values of leafy twigs were dictated by the ambient relative humidity rather than by height. Refilling of xylem vessels of apical leaves (branches) obviously mainly occurred via moisture uptake from the atmosphere. These findings could be traced back to the hydration and rehydration of mucilage layers on the leaf surfaces and/or of epistomatal mucilage plugs. Xylem vessels also contained mucilage. Mucilage formation was apparently enforced by water stress. The observed mucilage-based foliar water uptake and humidity dependency of the Pb values are at variance with the cohesion–tension theory and with the hypothesis that Pb measurements yield information about the relationships between xylem pressure gradients and height.

Sequential depolarization of root cortical and stelar cells induced by an acute salt shock – implications for Na+ and K+ transport into xylem vessels

Early events in NaCl-induced root ion and water transport were investigated in maize (Zea mays L) roots using a range of microelectrode and imaging techniques. Addition of 100 mm NaCl to the bath resulted in an exponential drop in root xylem pressure, rapid depolarization of trans-root potential and a transient drop in xylem K(+) activity (A(K+) ) within ∼1 min after stress onset. At this time, no detectable amounts of Na(+) were released into the xylem vessels. The observed drop in A(K+) was unexpected, given the fact that application of the physiologically relevant concentrations of Na(+) to isolated stele has caused rapid plasma membrane depolarization and a subsequent K(+) efflux from the stelar tissues. This controversy was explained by the difference in kinetics of NaCl-induced depolarization between cortical and stelar cells. As root cortical cells are first to be depolarized and lose K(+) to the environment, this is associated with some K(+) shift from the stelar symplast to the cortex, resulting in K(+) being transiently removed from the xylem. Once Na(+) is loaded into the xylem (between 1 and 5 min of root exposure to NaCl), stelar cells become more depolarized, and a gradual recovery in A(K+) occurs.

Effects of environmental parameters and irrigation on the turgor pressure of banana plants measured using the non‐invasive, online monitoring leaf patch clamp pressure probe

Turgor pressure provides a sensitive indicator for irrigation scheduling. Leaf turgor pressure of Musa acuminate was measured by using the so-called leaf patch clamp pressure probe, i.e. by application of an external, magnetically generated and constantly retained clamp pressure to a leaf patch and determination of the attenuated output pressure P(p) that is highly correlated with the turgor pressure. Real-time recording of P(p) values was made using wireless telemetric transmitters, which send the data to a receiver base station where data are logged and transferred to a GPRS modem linked to an Internet server. Probes functioned over several months under field and laboratory conditions without damage to the leaf patch. Measurements showed that the magnetic-based probe could monitor very sensitively changes in turgor pressure induced by changes in microclimate (temperature, relative humidity, irradiation and wind) and irrigation. Irrigation effects could clearly be distinguished from environmental effects. Interestingly, oscillations in stomatal aperture, which occurred frequently below turgor pressures of 100 kPa towards noon at high transpiration or at high wind speed, were reflected in the P(p) values. The period of pressure oscillations was comparable with the period of oscillations in transpiration and photosynthesis. Multiple probe readings on individual leaves and/or on several leaves over the entire height of the plants further emphasised the great impact of this non-invasive turgor pressure sensor system for elucidating the dynamics of short- and long-distance water transport in higher plants.

Oxygen Transport in Waterlogged Plants

In flooded soils, roots are exposed to a reducing environment with low oxygen availability. In order to supply roots with sufficient oxygen for respiration, most wetland plants form extended gas-filled cavities known as aerenchyma (in the root) or lacunae (in the shoot). Oxygen transport can be either diffusive or convective; according to general belief, the latter is restricted to the existence of separate “inlet” and “outlet” structures, e.g. in extended rhizomes. Mainly three separate mechanisms of convective oxygen transport have been identified so far: (1) Humidity-induced pressurization/convection that is driven by a difference in vapour pressure between the water-saturated gas phase of the leaf intercellulars and the microenvironment. When a significant overpressure in the gas phase of the leaf is maintained, gas flow to a distant “outlet”, being at atmospheric pressure, is generated. (2) Thermal osmosis, a mass flow of gas into the leaf driven by heat transfer in the opposite direction and (3) Venturi-induced convection, a gas flow initiated by an underpressure in the gas phase of broken culms (e.g. of Phragmites communis) that is generated by wind passing over them. Species with extended pathways for gas transport (with formation frequently being induced by hypoxia or anoxia) dominate in wetland communities and can even provide the rhizosphere of flooded soils with oxygen.

A non‐invasive probe for online‐monitoring of turgor pressure changes under field conditions

An advanced non-invasive, field-suitable and inexpensive leaf patch clamp pressure probe for online-monitoring of the water relations of intact leaves is described. The probe measures the attenuated output patch clamp pressure, P(p), of a clamped leaf in response to an externally applied input pressure, P(clamp). P(clamp) is generated magnetically. P(p) is sensed by a pressure sensor integrated into the magnetic clamp. The magnitude of P(p) depends on the transfer function, T(f), of the leaf cells. T(f) consists of a turgor pressure-independent (related to the compression of the cuticle, cell walls and other structural elements) and a turgor pressure-dependent term. T(f) is dimensionless and assumes values between 0 and 1. Theory shows that T(f) is a power function of cell turgor pressure P(c). Concomitant P(p) and P(c) measurements on grapevines confirmed the relationship between T(f) and P(c). P(p) peaked if P(c) approached zero and assumed low values if P(c) reached maximum values. The novel probe was successfully tested on leaves of irrigated and non-irrigated grapevines under field conditions. Data show that slight changes in the microclimate and/or water supply (by irrigation or rain) are reflected very sensitively in P(p).

Electrical signalling and cytokinins mediate effects of light and root cutting on ion uptake in intact plants

Nutrient acquisition in the mature root zone is under systemic control by the shoot and the root tip. In maize, exposure of the shoot to light induces short-term (within 1-2 min) effects on net K+ and H+ transport at the root surface. H+ efflux decreased (from -18 to -12 nmol m(-2) s(-1)) and K+ uptake (approximately 2 nmol m(-2) s(-1)) reverted to efflux (approximately -3 nmol m(-2) s(-1)). Xylem probing revealed that the trans-root (electrical) potential drop between xylem vessels and an external electrode responded within seconds to a stepwise increase in light intensity; xylem pressure started to decrease after a approximately 3 min delay, favouring electrical as opposed to hydraulic signalling. Cutting of maize and barley roots at the base reduced H+ efflux and stopped K+ influx in low-salt medium; xylem pressure rapidly increased to atmospheric levels. With 100 mm NaCl added to the bath, the pressure jump upon cutting was more dramatic, but fluxes remained unaffected, providing further evidence against hydraulic regulation of ion uptake. Following excision of the apical part of barley roots, influx changed to large efflux (-50 nmol m(-2) s(-1)). Kinetin (2-4 microM), a synthetic cytokinin, reversed this effect. Regulation of ion transport by root-tip-synthesized cytokinins is discussed.

Distribution and function of epistomatal mucilage plugs

Investigation of 67 gymnosperm and angiosperm species belonging to 25 orders shows that epistomatal mucilage plugs are a widespread phenomenon. Measurements of the leaf water status by using the leaf patch clamp pressure technique suggest that the mucilage plugs are involved in moisture uptake and buffering leaf cells against complete turgor pressure loss at low humidity.