Plant, Cell and Environment (1998) 21, 849–865

ORIGINAL ARTICLE OA 220 EN

Simultaneous recording of xylem pressure and trans-root

potential in roots of intact glycophytes using a novel xylem
pressure probe technique
L. H. WEGNER & U. ZIMMERMANN

Lehrstuhl für Biotechnologie der Universität, Biozentrum, Am Hubland, D-97074 Würzburg, Germany

ABSTRACT Key-words: Poaceae; Triticum aestivum L. cv. Alexandria;

wheat; Zea mays L. cv. Zelltic; maize; electrokinetic effects, pres-
The radial electrical potential difference between the root
sure-potential probe; trans-root potential (TRP); xylem pressure.
xylem and the bathing solution, i.e. the so-called trans-root
potential, was measured in intact maize and wheat plants
using a xylem pressure probe into which an Ag/AgCl elec- INTRODUCTION
trode was incorporated. Besides other advantages (e.g. Measurements of the electrical potential difference
detection and removal of tip clogging; determination of between the solution bathing the root and the xylem, i.e.
the radial root resistance), the novel probe allowed place- the so-called trans-root potential, provide a way to eluci-
ment of the electrode precisely in a single xylem vessel as date the contribution of electrical forces to solute and water
indicated by the reading of sub-atmospheric or negative transport through the root (Etherton & Higinbotham 1960;
pressure values upon penetration. The trans-root poten- Bowling & Spanswick 1964; Shone 1969; Davis &
tials were of the order of 0 to – 70 mV and + 40 to – 20 mV Higinbotham 1969; Läuchli, Spurr & Epstein 1971; Cortes
for 2- to 3-week-old maize and wheat plants, respectively. 1992; Rygol et al. 1993; Clarkson 1993; Marschner 1995).
Osmotic experiments performed on maize demonstrated Reliable data on these forces and their coupling to transpi-
that addition of 100 mM mannitol to the solution resulted ration-induced changes in xylem pressure, and to turgor
in a decrease of xylem pressure associated with a slow, but pressure of the hydraulically coupled root cells (Andrews
continuous depolarization. The depolarization was 1976; Stahlberg & Cosgrove 1995; Schneider, Zhu &
reversible upon removal of the mannitol. For wheat plants Zimmermann 1997a), are also urgently needed to explain
it could be shown that the oscillations of the xylem pres- the generation of radial turgor and osmotic pressure gradi-
sure described recently by Schneider et al. (1997, Plant, ents through the root cortex of transpiring plants (Rygol &
Cell and Environment 20, 221–229) were accompanied by Zimmermann 1990).
(rectangular, saw-tooth and/or U-shaped) oscillations in Solvent drag effects can account for many of these
the trans-root potential (but not by corresponding changes phenomena provided that the radial reflection coefficients
of the membrane potential of the cortical cells measured of the root are fairly low (Zimmermann et al. 1992; Rygol
simultaneously with conventional microelectrodes). et al. 1993). Values of the radial reflection coefficients
Increase of the light intensity (up to 550 µmol m–2 s–1) lower than σr = 1 were suggested by work on excised roots
resulted in a drop of the xylem pressure in wheat, whereas (Steudle 1989, 1992) and low-transpiring plants (Zhu et al.
the trans-root potential showed a biphasic response: first 1995). However, recent work on roots of high-transpiring
hyperpolarization (by about 10 mV) was observed, glycophytes has shown (Schneider et al. 1997a,b) that the
followed by depolarization (by up to about + 40 mV). radial reflection coefficients of roots of glycophytes can
Similar light-induced biphasic (but often less pro- assume values of up to σr = 1.
nounced) changes in the trans-root potential were also Even though this finding does not necessarily exclude
recorded for maize plants. Most interestingly, the the involvement of solvent drag effects in the transport
response of the trans-root potential was always faster (by properties of the root (see the analysis of water and solute
about 1–3 min) than the response of the xylem pressure transport through multiple membranes arranged in series;
upon illumination, suggesting that changes in the transpi- Curran & McIntosh 1962; Patlak, Goldstein & Hoffman
ration rate are reflected very quickly in the electrical 1963; Ginsburg 1971) alternative forces should certainly
properties of the root tissue. The impact of this and other be considered. The trans-root potential is a primary candi-
findings on long-distance transport of solutes and water date (see Katchalsky & Curran 1965; Rygol et al. 1993).
as well as on long-distance signalling is discussed. Our knowledge about the trans-root potential has been
reached from two different experimental approaches. Some
Correspondence: Prof. Dr U. Zimmermann. Fax: + 49 931 888 authors (Bowling & Spanswick 1964; Shone 1968, 1969;
4509; e-mail: [email protected] Davis & Higinbotham 1969; Dunlop & Bowling 1971a;
© 1998 Blackwell Science Ltd 849
850 L. H. Wegner and U. Zimmermann

De Boer, Prins & Zanstra 1983; Kennedy 1977; Vuletic & potential and its dependence on xylem pressure under vari-
Vucinic 1996) determined this parameter on excised roots ous environmental conditions.
by measuring the voltage drop between the exudate and the
bathing solution. Other authors (Dunlop & Bowling
MATERIALS AND METHODS
1971a,b; Bowling 1972; Dunlop 1973, 1982) recorded the
potential profile between the ‘xylem’ and the external Plant material
medium of intact plants by insertion of a microelectrode
Maize (Zea mays L. cv. Zelltic) and wheat (Triticum
into the root tissue.
aestivum L. cv. Alexandria) plants were cultivated as
The first approach suffers from two shortcomings. In
described before (Zhu et al. 1995; Schneider et al. 1997a).
excised roots the turgor and intracellular osmotic pressure
For experiments, 2- to 3-week-old plants were selected.
gradients collapse (see above, Rygol et al. 1993) indicating
The xylem pressure and trans-root potential measurements
that the behaviour of excised root systems may not always
were performed in the primary root. If not stated otherwise,
be a reliable guide to that of the intact plant (Clarkson
the roots were bathed in a standard solution containing
1993). Electrical double layers (Amin 1982) and longitudi-
2 mM CaCl2, 2 mM MgCl2, 1 mM KNO3 and 10 mM 2-[N-
nal potentials (Fensom 1957, 1962) which may be formed
Morpholino]ethane-sulfonic acid (MES); the pH was
in the lumen of the vessels (due to xylem loading, water
adjusted to 5·5 using 1,3-Bis tris-(hydroxymethyl)-methy-
ascent and/or ion accumulation in the leaf vessels, e.g.
lamino]propane (BTP). The osmolality was determined to
Schwenke & Wagner 1992; Canny 1990, 1993, 1995;
be 17 mosmol kg–1 (Osmomat 030, Gonotec, Berlin,
Wegner & Raschke 1994) may change upon excision (see
Germany). In most cases, experiments were performed
also Ginsburg 1972).
under laboratory conditions (temperature 20–24 °C, rela-
The second approach must also be criticized because
tive humidity 20–50%; illumination 10 µmol m–2 s–1). For
there is no evidence that the placing of the innermost
additional illumination a sodium lamp (Adolf Schuch KG,
microelectrode was inside a vessel (without disrupture of
Worms, Germany) was used; the irradiance level was
the water column). Furthermore, clogging of the micro-
changed by variation of the distance between the lamp and
electrode tip by cellular debris during impalement through
the leaves.
the root tissue can lead to erroneous potential recordings
(Dunlop 1982), and damage of the tissue may result in a
(partial) short-circuit between xylem and medium (see also Xylem pressure-potential probe
Anderson & Higinbotham 1975).
The probe consisted of a xylem pressure probe in which a
In the light of these uncertainties associated with previ-
potential-measuring Ag/AgCl electrode was integrated.
ous trans-root potential measurements, it is perhaps not
The construction, principles and pitfalls of the xylem pres-
surprising that the trans-root potential data conflicted with
sure probe, as well as the probing procedure for vessels are
results obtained by other methods. For example, K+-ion
described elsewhere (Balling & Zimmermann 1990).
gradients in the root tissue as revealed with the electron-
Incorporation of the electrode required some modifica-
probe microanalysis technique (Läuchli & Epstein 1971;
tions of the probe (Fig. 1a).1 The microcapillary of the
Dunlop & Bowling 1971a) could not be detected by suc-
probe (length about 30 mm) was made from borosilicate
cessive radial insertion of a K+-selective microelectrode
glass (outer diameter 1 mm, inner diameter 0·5 mm;
into the root tissue. Furthermore, measurements of trans-
Hilgenberg GmbH, Malsdorf, Germany). It was connected
root potentials lead to conflicting data concerning the role
via a pressure-tight seal to a cylindrical hole (length
of stelar proton pumps (Dunlop 1973; Dunlop & Bowling
28 mm; diameter 1 mm) drilled into the perspex chamber
1971b; De Boer et al. 1983).
of the probe. This hole contained the Ag/AgCl electrode
In this communication we demonstrate that the problems
(length 20 mm; diameter 0·2 mm). About 4 mm of the tip
of the microelectrode technique mentioned above can be
of the silver wire was chlorinated and subsequently coated
overcome by incorporation of a potential-recording
galvanically with silver alginate; the remaining part was
Ag/AgCl wire into the xylem pressure probe (Balling &
covered with a thin Teflon layer. The chlorinated part of the
Zimmermann 1990; Schneider et al. 1997a,b). Such a
wire was positioned close to the pressure tight seal
xylem pressure-potential probe has the advantages (1) that
between the microcapillary and the perspex chamber.
a proper probing of a vessel is indicated by the pressure
Electric contact between the xylem sap and the electrode
signal, (2) that clogging of the microcapillary tip can easily
was achieved by filling the capillary and the horizontal
be removed by the displacement of the metal rod within the
hole with degassed 50 mM KCl solution.
pressure probe and (3) that the integrated electrode in the
probe can be used to determine separately the radial elec-
1
trical resistance of the root (which is an indicator for a Note that the xylem pressure-potential probe developed here is
short-circuit of the tissue). completely different from the probe described by Hüsken, Steudle
& Zimmermann (1978) for cell turgor measurements in cells of
Control experiments (including simultaneous membrane higher plants. The probe of Hüsken et al. can be used (but only
potential measurements in cortical cells) as well as applica- under a very few circumstances) for the measurement of above-
tion of this novel pressure probe to maize and wheat roots atmospheric pressure values; it also requires the use of oil-filled
showed that reliable data can be obtained on the trans-root capillaries.

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 851

Figure 1. Schematic diagrams of (a) the xylem pressure probe containing the integrated trans-root potential-measuring electrode and (b) the
electronic circuit used for measuring the radial root potential and resistance. 1 = microcapillary; 2 = perspex chamber; 3 = pressure
transducer; 4 = metal rod; 5 = micrometer screw; 6 = pressure-tight rubber seals; 7 = Ag/AgCl electrode. For further explanations, see text.

The vertical hole (length 39 mm; diameter 1 mm) con- (length 25 mm; diameter 0·7 mm) and a salt bridge filled
nected the pressure transducer and the metal rod (for injec- with 50 mM KCl and 2% Agar.
tion of pressure pulses into the punctured vessel; see Tip potentials were virtually zero due to the relatively
Balling & Zimmermann 1990). This hole was filled with large tip diameter of the microcapillary. Liquid junction
deionized and degassed water. Experiments showed that potentials were also negligible because of the very similar
deionized water was required in order to prevent corrosion mobilities of K+ and Cl–.
of the pressure transducer and to establish a high resistance In contrast to conventional electrophysiological set-
(of about 1 MΩ) in order to avoid electrical interference ups (Fig. 1b), the xylem electrode was grounded and the
between the electrode and the transducer. reference electrode was connected to a high impedance
Problems arose from diffusion of KCl into the liquid of differential amplifier (input resistance 1012 Ω). Under
the vertical hole. However, the Teflon coating of the elec- these conditions interference between the pressure trans-
trode prevented a drift in the potential for at least 10 h. ducer and the potential-measuring electrode could be
This time was more than sufficient to perform an experi- eliminated. This procedure required that any contact of
ment. Before each new experiment the electrolyte solu- the bath solution and of the plant with ground was
tion in the horizontal hole was exchanged by fresh avoided. Control experiments, in which the pressure
solution; additionally, the probe was washed and com- transducer was disconnected and the reference electrode
pletely refilled every 2–3 d. The reference electrode in was grounded, revealed no differences in the potential
the solution consisted of a thick chlorinated Ag wire readings (data not shown).
© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
852 L. H. Wegner and U. Zimmermann

Despite these differences in recording, the values of the and plant experiments (Balling & Zimmermann 1990;
trans-root potentials given below are corrected according Benkert, Balling & Zimmermann 1991; Zimmermann
to convention (i.e. by reversing, as if the potential of the et al. 1995a; Zimmermann, Meinzer & Bentrup 1995b;
solution had been set to zero). Schneider et al. 1997b). The modification of the probe for
In addition to the trans-root potential, the setup could also simultaneous measurements of the xylem pressure and the
be used to estimate the radial electrical resistance of the root trans-root potential required additional verifications of the
tissue (Fig. 1b). To this end, a pulse generator (TE 7702, proper reading of these two parameters by control and
Toellner, Frankfurt, Germany) was connected to the solution model experiments.
via an Ag wire (length 15 mm; diameter 0·7 mm). This led to An important question was whether the design and the
a short-circuit of the pressure-potential probe and the bath. arrangement of the xylem pressure-potential probe resulted
Square pulses of 1·25 s duration and of different amplitudes in interferences between pressure and potential recordings.
were injected, and the corresponding changes in the appar- This was tested by connecting the chamber of the grounded
ent trans-root potential were recorded using the probe elec- probe via an ordinary glass capillary with a conventional
trode (see below). The magnitude of the current pulse was Ag/AgCl electrode (filled with a degassed 50 mM KCl
determined by measuring the voltage drop across a 4·7 kΩ solution; Fig. 2a). This electrode was connected to a volt-
resistance (arranged in series with the electrode) using a dif- age source which allowed the stepwise application of volt-
ferential amplifier (input resistance 1012 Ω). In order to pre- ages with increasing amplitude.
vent artifacts arising from electrode polarization, only small The glass capillary (length 60 mm, internal diameter
currents were applied (less than 1 µA). From these current 0·5 mm) was melted at half-length and thinned by pulling
and voltage measurements, the radial root resistance could to a few micrometers in diameter. The part of the glass cap-
be calculated provided that the resistance of the electrode illary facing the probe was filled with a degassed 50 mM
was taken into account and was measured before placing the KCl solution, the other one with a degassed 1 mM KCl
probe into a vessel. The calculations gave a good estimate of solution. Thus conditions were as similar as possible to
the radial root resistance, provided that the conductivity of those which existed after in situ probing of a xylem vessel
the xylem sap and the bath solution were of comparable containing low concentrations of KCl (or other elec-
order and large in relation to that of the root tissue. Control trolytes). In the first set of experiments rectangular or
experiments showed that this assumption was fulfilled under (nearly) sinusoidal pressure pulses of sub-atmospheric (0
the experimental conditions. to + 0·1 MPa)2 and above-atmospheric (+ 0·1 MPa
An additional requirement for resistance measurements to + 0·6 MPa) amplitude were injected into the fluid of the
was that the resistance of the electrode was not changed due assembly by appropriate displacement of the metal rod. As
to clogging during insertion through the root tissue. indicated in Fig. 2b, the Ag/AgCl electrode did not read
Clogging occurred occasionally and was detected by routine any signal from the pressure transducer. Even at a voltage
injection of pressure pulses into the punctured vessel (by of – 50 mV, no effect of the pressure pulses on the voltage
appropriate displacement of the metal rod within the probe); recording was observed (data not shown). Similarly, the
clogged tips showed characteristically large, persistent pres- pressure recording was not affected by a voltage offset
sure excursions. Such measurements were discarded. imposed on the recording electrode (Fig. 2c). Experiments
The resistance measurements were performed at con- in which a sub-atmospheric pressure (0·04 MPa) or above-
stant xylem pressure, i.e. at water equilibrium, because this atmospheric pressure (0·25 MPa) was established yielded
condition minimized possible artifacts arising from con- similar negative results (data not shown). This indicated
centration changes at the tip of the microcapillary due to that interferences between voltage and pressure recording
KCl leakage or flux of xylem sap into the capillary. could be excluded.
The second question was whether streaming potentials
and/or changes in the tip potential could occur upon
Cellular membrane potentials changes in the xylem pressure of the probed vessel due to
Ag/AgCl electrodes filled with 1 M KCl were used. The influx of low-conductivity xylem sap into the microcapil-
resistance was about 300 MΩ. The small tip potentials lary. Besides such effects, KCl leakage from the microcap-
were not taken into account (Blatt 1991). The membrane illary into the vessel (particularly at low transpiration rates
potential difference of wheat cortical cells of the outermost and/or during long-term measurements) can be envisaged
layer was determined in the conventional way using a dif- which may affect xylem sap composition and, in turn, the
ferential amplifier (Zimmermann, Büchner & Benz 1982). trans-root potential (see ‘Materials and methods’).
In order to test if such effects could lead to erroneous
trans-root potential recordings, positive or negative
RESULTS volume increments (10 and 5 nL, respectively) were
injected into or sucked out of a probed root vessel of wheat
Evaluation of the method
by appropriate displacement of the metal rod (Fig. 3a). In
The ability of the xylem pressure probe to accurately mea-
sure negative pressure values in conducting vessels of 2
Note that all pressure (tension) values are quoted as absolute
higher plants and tall trees has been tested in many model pressures (tensions; atmospheric pressure = + 0·1 MPa).

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 853

Figure 2. (a) Set-up used in order to exclude interference of rectangular or ‘sinusoidal-shaped’ pressure signals with the probe-electrode (b)
and of rectangular voltage signals with the pressure transducer (c). The pressure signals in (b) were generated by appropriate displacement of
the metal rod shown in Fig.1a; the voltage signals were generated by a voltage source arranged in series with the conventional electrode in (a).
For further details of these model experiments, see text.

order to lower the probability of cavitation, the shoot and Positive, but not negative volume increments caused a tran-
the leaves of the plant were wrapped in a plastic bag before sient hyperpolarization. As shown in Fig. 3a, the pressure
insertion of the probe. In non-transpiring plants, the xylem changes dissipated within seconds, indicating that the tip of
pressure of the root is normally clamped at positive, sub- the microcapillary was not clogged (Benkert et al. 1991).
atmospheric values (Zhu et al. 1995). The trans-root potential, however, relaxed back to the initial
The injection of the volume increments was performed value significantly slower (i.e. within about 1 min).
after reading of a nearly constant xylem pressure (about Formation of a gas bubble (arrow ‘B’ in Fig. 3a) resulted
+ 0·05 MPa) and trans-root potential (about –2 mV). in a change of the trans-root potential to more negative
© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
854 L. H. Wegner and U. Zimmermann

Figure 3. ‘In vivo’ (a) and ‘in vitro’ (b) model experiments performed in order to prevent streaming potentials and/or changes in the tip
potential due to pressure-induced volume flow through the tip of the microcapillary of the pressure-potential recording probe of Figure 1. (a)
Trans-root potential (TRP; upper trace) and xylem pressure (Px, lower trace) measurements in a probed root vessel of an 18-d-old intact, non-
transpiring wheat plant. Probing of a vessel is indicated by a drop of the pressure into the sub-atmospheric range (arrow ‘A’). After reading of
a constant xylem pressure (about + 0·05 MPa) and trans-root potential (about – 2 mV), positive and negative volume increments were injected
into or sucked out of the vessel (by means of the metal rod in Figure 1). The rapid dissipation of the induced pressure changes demonstrated
that the tip of the microcapillary was not clogged. Positive, but not negative pressure spikes were associated with a change in the trans-root
potential of about 1 min duration. Occurrence of a gas bubble (arrow ‘B’) resulted in a drop of the trans-root potential to about – 20 mV.
Subsequent injection of positive volume increments (arrows ‘C’) did not lead to pressure spikes (due to the high compressibility of the gas
bubble). However, transient hyperpolarization spikes were still generated which lasted as long as the spikes observed in the absence of a gas
bubble. (b) Injection of positive volume increments into a 1 mM KCl solution resulted in a transient pressure increase within the probe (P;
lower trace) associated with a small (capacitive) hyperpolarization spike (V; upper trace). Both spikes disappeared within seconds. This
control experiment shows that the hyperpolarization spikes in (a) are presumably generated by the plant.

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 855

values. Subsequent injections of positive volume incre- through the tissue. In order to test this possibility, the micro-
ments (arrows ‘C’ in Fig. 3a) resulted in only very small capillary was pushed radially through the root until the tip
pressure changes (due to the high compressibility of the reached the opposite side of the bath solution. As a rule, this
gas bubble; Benkert et al. 1991), but the hyperpolarization led to (partial) clogging of the tip. Then, positive pressure
spikes were still observed. These hyperpolarization spikes pulses were injected by displacement of the rod in analogy
presumably did not arise from changes in the tip potential with the removal of clogging material from the tip during
or from streaming potentials. This was suggested by a xylem pressure measurements. Again, no change of the elec-
control experiment in which the microcapillary of the trode potential was observed (data not shown), suggesting
probe was dipped into a 1 mM KCl solution under atmo- that the transient hyperpolarization caused by pressure
spheric pressure (Fig. 3b). Upon injection of a volume pulses after puncture of a vessel (see also below) was not
increment into the solution only a relatively small (pre- due to a removal of debris from the microcapillary tip.
sumably capacitive) potential spike was observed which Another important question was also whether impale-
dissipated as rapidly as the pressure spike. ment of a vessel by the probe short-circuited the xylem
It appears therefore that the hyperpolarization spikes with its surrounding (the apoplast of the cortex and the
observed in the probed vessel of the wheat root (Fig. 3a) ambient medium) due to local mechanical destruction of
arose from transient changes in the electrostatics of the the cells. Such an effect would lead to an underestimation
xylem walls (Amin 1982) or from the occurrence of action of the trans-root potential. An answer to this question was
potentials in response to changes in pressure (Stahlberg & provided by measurements of the radial resistance of the
Cosgrove 1992). Pressure-induced action potentials have root before and after probing of a vessel (see ‘Materials
occasionally been observed in higher plants and have also and methods’). Typical recordings of the current pulses
been reported for giant algal cells (see e.g. Zimmermann & and the corresponding changes in potential are shown in
Beckers 1977). Fig. 4. In order to perform resistance measurements,
An important result of these studies was that no long- recordings of the trans-root potential were interrupted by
term changes of the trans-root potential were observed connecting and subsequently disconnecting the pulse gen-
(Fig. 3a) even after the injection of 10 nL of the 50 mM KCl erator, as indicated by the arrows. Note that the same
solution into the probed vessel of a non-transpiring plant. potential was measured before and after application of the
Another potential source of error was contamination of current pulses, indicating that the tissue was not damaged
the tip by cell debris or cell wall material during penetration by this procedure.

Figure 4. Part of an experiment in which the radial resistance of the root of a 19-d-old wheat plant was determined using the xylem pressure-
potential probe after switching the electronic circuit of Figure 1b from trans-root potential to resistance measurements (see ‘Materials and
methods’). A vessel of a root (23 cm in length) was probed 18 cm away from the tip using a microcapillary filled with 1 M KCl. The resistance
was measured by injection of rectangular 1·5 s current pulses (lower traces) into the bath and by simultaneous recording of the voltage response
between the electrodes (upper traces). The arrows indicate the start and the end of the resistance measurements. Note that the scaling bars only
refer to the resistance measurement; voltage recordings before and after the application of the pulse are therefore given directly at the trace.
Calculation of the resistances and comparison of the data measured before (left-hand side) and after (right-hand side) vessel probing (see
‘Materials and methods’) show that the resistance increased from 2·9 to 6·8 MΩ after probe insertion. This increase was attributed to the radial
resistance of the root tissue. During the trans-root resistance measurement, the xylem pressure was – 0·03 MPa (trace not shown).

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
856 L. H. Wegner and U. Zimmermann

Generally, the radial resistance of the root was remark- those of maize roots. This finding was interesting because
ably high, but the scatter of the data was quite large. For the root of wheat is thinner (only 4 cortical layers) than that
maize plants the values ranged from 5 to 70 MΩ when of maize (9 cortical layers). It demonstrated (in addition to
using microcapillaries filled with 50 mM KCl (When the the high resistance values) that it was very unlikely that the
tip was dipped into standard solution, capillary resistance roots were short-circuited by the impalement procedure.
was 23·5 ± 6·5 MΩ; n = 10). Such variations are not
uncommon for plant cells (Wang et al. 1991) and may
Trans-root potentials of maize
reflect physiological features. Interestingly, the values
measured here were much higher than those reported for A typical recording of the xylem pressure-potential probe in
other tissues as well as for the longitudinal resistance of a maize root bathed in standard solution is shown in Fig. 5.
maize roots (Anderson & Higinbotham 1976). The reason Probing of a xylem vessel was indicated by a rapid drop of
for this was presumably the endodermis which formed a the pressure from slightly above-atmospheric to positive
tight seal around the microcapillary after positioning in a sub-atmospheric values of about + 0·04 MPa (arrow ‘A’ in
xylem vessel. Fig. 5). This value agreed well with those measured previ-
In maize, somewhat lower values of 1·1 ± 0·2 MΩ (n = 5) ously in the xylem of roots bathed in nutrient solution (Zhu
were found when the tip of the microcapillary was filled et al. 1995; Schneider et al. 1997a). During penetration of
with 1 M KCl. The use of the high electrolyte concentration the microcapillary through the root tissue fluctuations in the
improved the accuracy because of the reduction of the tip trans-root potential of up to – 50 mV were recorded. At the
resistance (1·3 ± 0·6 MΩ, n = 5). However, at high KCl con- instant of vessel probing (arrow ‘A’ in Fig. 5) the potential
centrations, salts may accumulate in the apoplast of the dropped and assumed a stable value of about – 10 mV. The
stele due to KCl leakage from the tip of the microcapillary tip was not clogged by cell wall debris, as verified by injec-
into the vessel. This would lead to an underestimation of the tion of a positive pressure pulse via appropriate displace-
actual resistance of the root. For wheat, tips of smaller ment of the metal rod (arrow ‘B’). The pressure relaxed
diameter could be used (tip resistance 2·8 ± 0·6 MΩ, n = 5). back within seconds to the value recorded before the pulse
Therefore, the leakage problem may be less severe. The was applied. Switching on the bath perfusion with standard
resistance of wheat roots was on average 3·2 ± 0·9 MΩ solution (arrow ‘C’ in Fig. 5) created spikes in the trans-root
(n = 5) at 1 M KCl. Comparison of the data indicates that the potential, but did not affect the average values of the trans-
resistance values of wheat roots were always higher than root potential and the xylem pressure.

Figure 5. Simultaneous recording of xylem pressure (Px; lower trace) and the trans-root potential (TRP; upper trace) in a primary root (length
18 cm) of a 15-d-old maize plant at a light intensity of 10 µmol m–2 s–1, a temperature of 22 °C and a relative humidity of 42%. The roots were
bathed in standard solution (the osmolality was determined to be 22 mosmol kg–1). The probe was inserted into a vessel about 16 cm away from
the cap (time ‘zero’). Puncture of the vessel is indicated by a sudden decrease of the pressure from atmospheric to + 0·04 MPa (arrow ‘A’). At this
instant a nearly constant trans-root potential of – 10 mV was recorded. After about 10 min, a positive pressure pulse was applied which was
associated with a hyperpolarization spike (arrow ‘B’; see Figure 3a). The onset of the perfusion of the bath with standard solution (arrow ‘C’) was
usually accompanied by a few voltage spikes. Replacement of the standard solution by a 131 mM mannitol solution (arrow ‘D’; final osmolality
153 mosmol kg–1) resulted in a decrease of the xylem pressure to – 0·04 MPa, followed by a backregulation to a slightly less negative pressure
value. Upon addition of the mannitol solution the trans-root potential hyperpolarized transiently before depolarization was observed. Some time
after a steady-state potential of about 0 mV was reached, the osmolality of the perfusion medium was reduced (arrow ‘E’; final osmolality 38
mosmol kg–1). Lowering of the osmolality resulted in a pressure increase associated with a hyperpolarization.

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 857

Adding 131 mM mannitol to the bath (arrow ‘D’ in in nutrient medium and behave like a perfect osmometer
Fig. 5, final osmolality 153 mosmol kg–1) resulted in a even at low rates of transpiration (Schneider et al. 1997a).
rapid decrease of the xylem pressure down to a value of Very frequently, the xylem pressure did not attain a constant
– 0·05 MPa, followed by a slight backregulation of the value at high light intensity or after addition of an
pressure to less negative values.3 In agreement with previ- osmoticum. In contrast, oscillations in xylem pressure were
ous findings on low-transpiring maize plants (Zhu et al. observed (Schneider et al. 1997a) which were apparently
1995; Schneider et al. 1997a) the root did not behave as a correlated with periodic changes of the stomatal conduc-
perfect osmometer upon addition of the mannitol solution. tance (Raschke 1965). The xylem pressure and trans-root
Calculation of the radial reflection coefficient for manni- potential measurements revealed similarities, but also differ-
tol from the osmotically induced xylem pressure change ent features of wheat roots compared to maize roots.
yielded a value of about 0·2. Upon addition of the Typical recordings of both parameters measured on
osmoticum, the trans-root potential responded in a bipha- wheat roots bathed in standard solution are shown in Fig. 6.
sic manner. After a rapid hyperpolarization to about As expected, the xylem pressure assumed negative
– 20 mV the trans-root potential depolarized to about values. On average, a xylem pressure was recorded of
0 mV within 25 min. Interestingly, the depolarization –0·038 ± 0·061 MPa (n = 22; light intensity about 10 µmol
started before the xylem pressure had reached its most m–2 s–1). The trans-root potential was positive (Fig. 6a–c)
negative value (Fig. 5). The osmotic effect on the xylem or close to zero (Fig. 6d). The mean steady state value of
pressure and the trans-root potential was almost com- the trans-root potential was + 5·8 ± 10·5 mV (n = 22).
pletely reversible. When the mannitol solution was Oscillations of the xylem pressure were observed regu-
replaced by the standard solution (arrow ‘E’), the original larly. As shown in Fig. 6a, the amplitude of the oscillations
values of both parameters were restored within 25 min. was strongly enhanced by an increase in light intensity to
The establishment of a negative pressure during the 250 µmol m–2 s–1. Upon illumination the xylem pressure
osmotic treatment of the roots indicated that tiny gas bub- decreased to more negative values. Thereafter, the mean
bles (e.g. adhering to the Ag/AgCl electrode) were not pre- value of the xylem pressure shifted progressively to more
sent. Experiments in which negative pressures could not be positive values, and there was a decrease in the amplitude
established by osmotic stress were discarded. of the oscillations. This damping phenomenon was also
It should also be noted that the transient hyperpolariza- observed when light intensity was kept constant at
tion response of the trans-root potential upon the addition 10 µmol m–2 s–1 (see e.g. Fig. 6b or d).
of mannitol was only observed in part of the experiments, The trans-root potential responded to the oscillations in
whereas the (reversible) depolarization response always xylem pressure. The shape of the trans-root potential var-
occurred. The average amplitude of the osmotically ied from plant to plant. Four response patterns were
induced depolarization response was 10 ± 5 mV (n = 7). observed which are presented in Fig. 6a–d. In Fig. 6a, the
Recordings on roots of other maize plants yielded similar trans-root potential increased rapidly by 10–40 mV once
results. The trans-root potential varied between – 70 mV the pressure had reached a minimum, and decreased
and – 10 mV (n = 60). In three cases, a slightly positive accordingly at a pressure maximum (see magnification of
potential of 5 mV was recorded. The average trans-root one of the oscillation cycles in the inset in Fig. 6a).
potential was – 20 mV. No correlation was found between Between the two pressure extremes, the trans-root poten-
the value of root resistance (measured routinely in each tial remained almost stable; the oscillations in the trans-
experiment, see above) and the corresponding value of the root potential apparently assumed a ‘rectangular shape’.
trans-root potential. Such a correlation would be expected if During the first part of the experiment the mean value of
leakage problems within the root tissue (due to damage of the trans-root potential shifted to more positive values.
cells and/or of the endodermis, etc.) had played any role. After about 3 h the mean value decreased, however, contin-
uously. After about 5 h, the value recorded at the low light
intensity was approximately reached. The amplitude of the
Trans-root potentials of wheat
oscillations in trans-root potential decreased progressively
In the following figures trans-root potential recordings per- in a manner which corresponded to the changes in xylem
formed on roots of intact wheat plants are shown. Wheat pressure.
roots exhibit slightly negative xylem pressures when bathed In Fig. 6b, a light intensity of 250 µmol m–2 s–1 was
selected. It can be seen that the oscillations in xylem pres-
3
It should be noted that a stable pressure was not always reached. sure were more regular within the first hours after vessel
Instead, the xylem pressure increased again in order to assume a probing than in Fig. 6a. However, (as already mentioned
slightly less negative (or more positive) value. This backward reg- above), the average xylem pressure also shifted with time
ulation phenomenon of xylem pressure (which very often preceded to slightly less negative values combined with a decrease
regular oscillations in wheat) is well-known from previous studies of the amplitude of the oscillations. By analogy to the mea-
on glycophytes (Zhu et al. 1995; Schneider et al. 1997a). The
‘overshoot’ effect may be linked with transient stomatal closure
surements in Fig. 6a, depolarization of the trans-root
and/or with the water exchange between the vessels and the potential occurred nearly instantaneously once the xylem
hydraulically coupled tissue cells (see also in the context, pressure had reached its minimum value. Then, however,
Brogårdh, Johnsson & Klockarde 1974; Steudle 1992). the trans-root potential did not remain constant; it relaxed
© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
858 L. H. Wegner and U. Zimmermann

back continuously to lower values. An instantaneous Occasionally, changes in the trans-root potential
hyperpolarization at the pressure maximum as found in the occurred only during the phase of xylem pressure decrease
experimental run of Fig. 6a was not observed. The changes (Fig. 6c). The response of the trans-root potential assumed
in the trans-root potential assumed a ‘sawtooth’ pattern. a ‘U-shape’, i.e. the potential decreased, passed through a
The depolarization response was correlated with the minimum and increased again. In contrast to Fig. 6b, the
minimum in xylem pressure during the whole experiment, average trans-root potential drifted towards more positive
but its magnitude decreased continuously, closely related values (from about + 20 mV to about + 40 mV) during the
with the decrease of amplitude of the xylem pressure course of the experiment.
oscillations. Simultaneously, after about 2 h the average Frequently, fluctuations of trans-root potentials were
trans-root potential assumed more negative values. measured (Fig. 6d) which showed components of the three

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 859

Figure 6. Simultaneous recordings of oscillations in the xylem pressure (Px) and in the trans-root potential (TRP) in the roots of 9-18-d-old
wheat plants. Measurements were performed in standard solution at a temperature of 22–28 °C, a relative humidity of between 20 and 50% and a
light intensity of 10 µmol m–2 s–1, except that in Fig. 6a the light intensity was increased to 250 µmol m–2 s–1 (arrow ‘B’) 45 min after insertion of
the probe (arrow ‘A’) and that in Fig. 6b the light intensity was adjusted to this value throughout the experiment. (a) Rectangular-shaped
oscillations of the trans-root potential. Conditions: 9-d-old plant, impalement 27·8 cm away from the cap, root length 29 cm. Inset: part of the
experiment is shown at larger magnification in order to demonstrate the correlation between the extremes in xylem pressure and the
depolarization/hyperpolarization response of the trans-root potential. (b) ‘Sawtooth-shaped’oscillations of the trans-root potential. Conditions:
10-d-old plant, impalement 13 cm away from the cap, root length 21 cm. (c) U-shaped oscillations of the trans-root potential. Conditions: 18-d-
old plant, impalement 13·2 cm away from the cap, root length 16 cm. (d) Oscillations of the trans-root potential composed of elements of (a)–(c).
Conditions: 16-d-old plant, impalement 11·3 cm away from the cap, root length 17 cm. For further explanations, see text.

© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
860 L. H. Wegner and U. Zimmermann

response patterns described in Fig. 6a–c. Quite often, volt- in which xylem pressure, trans-root potential and mem-
age recordings were noisy when the trans-root potential brane potential were recorded in parallel. In this experi-
fluctuated in a U-shaped manner (Fig. 6d). ment, the oscillations in xylem pressure were
These examples demonstrate that the response pattern of accompanied by regular, U-shaped oscillations of the
the trans-root potential upon xylem pressure oscillations was trans-root potential as depicted in Fig. 6c. It is obvious
quite variable. The experiments illustrated in Fig. 6 as well that these oscillations were not reflected in the mem-
as in 19 other experiments showed that there was apparently brane potential of the outermost cortical cells. The mag-
no correlation between the pattern of the trans-root potential nitude of the cellular potential (– 110 to – 130 mV) was
response and the initial xylem pressure value recorded even slightly more negative than that recorded in control
immediately after positioning the microcapillary within experiments (– 102 ± 8 mV; n = 6). Similar values for
the vessel. wheat roots were reported by Walker & Graham (1987).
However, there were some common features in the The cellular potential in Fig. 7 drifted during 2 h slightly
response pattern of the trans-root potential upon xylem to more negative values, but a similar effect was also
pressure oscillations. (1) The depolarization of the trans- seen occasionally in control experiments without the
root potential was almost always correlated with a mini- inserted probe (data not shown).
mum and correspondingly the hyperpolarization very
frequently with a maximum in xylem pressure. (2)
Light-induced changes in the trans-root potential
Pressure oscillations and trans-root potential changes had
and xylem pressure
the same period, i.e. 34·3 ± 11·7 min (n = 23). The maxi-
mum amplitude of the trans-root potential changes did not In the light of the above and other experiments it is
depend on the type of ‘oscillation’. The average value was unlikely that the variable response of the trans-root poten-
determined to be 16·6 ± 8·0 mV. The standard deviation tial upon xylem pressure oscillations arose from damage
shows that the amplitude was subjected to relatively large of the root tissue. The results rather indicated that xylem
fluctuations. pressure and trans-root potential are linked with each
In the following we investigated whether the oscilla- other. This was also suggested by experiments in which
tions in the trans-root potential were accompanied by the effect of increased light intensity on xylem pressure
corresponding changes in the membrane potential of cor- and trans-root potentials was investigated in more detail.
tical cells. Because of technical reasons the membrane As shown in Fig. 8a for wheat, an increase of the light
potential of cortical cells of the outermost layer could intensity from 10 to 250 µmol m–2 s–1 (arrow) resulted
only be measured. To this end, a microelectrode was after a delay of about 3 min (i.e. the time the lamp required
inserted into a cell located near the insertion point of the to emit maximum light intensity) in a decrease of the
pressure-potential probe. Figure 7 shows a measurement xylem pressure (within about 15 min) from + 0·03 MPa

Figure 7. Part of an experiment in which the membrane potential (lower trace) of a cortical cell in the outermost layer of a root of an 18-d-
old wheat plant was recorded together with the xylem pressure (central trace) and the trans-root potential (upper trace). The cortical cell was
impaled 97 min after the insertion and close to the insertion point of the modified xylem pressure probe into a vessel (about 7 cm away from
the cap of a 18 cm long root). Conditions: standard solution, relative humidity 18·5%, temperature 26 °C and light intensity 10 µmol m–2 s–1.
Note that the oscillations in xylem pressure and the U-shaped oscillations in the trans-root potential were not reflected in the membrane
potential of the cortical cell.

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 Xylem pressure and trans-root potential 861

In these experiments (Fig. 8), the light intensity was var-

ied between 170 and 550 µmol m–2 s–1, leading corre-
spondingly to an increase in the xylem pressure changes
(average value of the maximum xylem pressure decrease
upon illumination: – 0·20 ± 0·08 MPa). However, a corre-
lation of the hyper-/depolarization response of the trans-
root potential with light intensity (or xylem pressure) could
not be detected. The biphasic response of the trans-root
potential was apparently independent of the light intensity
in the range investigated here. Only in one experiment
hyperpolarization, but no subsequent depolarization was
observed.
The hyperpolarization response upon illumination always
preceded the response of xylem pressure by one to three
minutes. The minimum value of the trans-root potential was
reached after about 8 min (mean value; time intervals rang-
ing from 100 s to 1300 s, n = 19), thus on average about
9 min before the minimum xylem pressure was established
(on average, pressure passed through a minimum after
17 min; time intervals ranged from 300 to 2500 s, n = 16).
When the light intensity was reduced to laboratory level
(10 µmol m–2 s–1) again, the opposite effects were often
recorded: a rapid, transient depolarization by 4–26 mV
(mean value: 11 mV, n = 8) occurred in most experiments,
followed by a hyperpolarization to a final value that was by
10–40 mV (mean value 21 mV) more negative than that
measured at a high light intensity (data not shown).
However, it must be noted that in three experiments no
steady hyperpolarization was observed. Xylem pressure
Figure 8. Typical recordings of the effects of illumination on the
responded with a delay of several minutes and reached a
xylem pressure (Px; upper trace) and the trans-root potential
(TRP; lower trace) in the root xylem of a 13-d-old wheat plant (a) new, more positive level within about 30 min (data not
and of a 14-d-old maize plant. The arrows mark the instance at shown).
which the light intensity was increased from 10 µmol m–2 s–1 to Qualitatively similar observations were made on maize
250 µmol m–2 s–1. Conditions: standard solution, relative humidity roots. An example is shown in Fig. 8(b). When the plant
35%, temperature 18 °C (maize) and 22 °C (wheat). Note that the was exposed to the same light regime as mentioned above,
response of the trans-root potential preceded the response of the xylem pressure responded with a lag time of about 2 min;
xylem pressure upon illumination.
then a decrease from + 0·065 MPa to – 0·172 MPa was
observed followed by a backregulation to about
down to – 0·12 MPa. In contrast, the trans-root potential – 0·07 MPa (see Zhu et al. 1995; Schneider et al. 1997a
dropped almost immediately (within 60 s) after switching and footnote 3) In contrast to the pressure response on illu-
on the light.4 A minimum value of – 18 mV was recorded mination, hyperpolarization was observed within 30 s; the
after about 8 min, i.e. about 10 min before the xylem trans-root potential dropped by 15 mV to a minimum value
pressure had reached its minimum value. Then, depolar- of – 30 mV and depolarized then to – 21 mV. In this exper-
ization of the trans-root potential was observed. After iment, the potential drifted to more negative values again
about 20 min an almost stable potential of + 7 mV was after the backregulation of the xylem pressure was com-
established. pleted. The minimum in trans-root potential preceded that
On average, the maximum amplitude of the transient of the xylem pressure by about 6 min.
hyperpolarization, following an increase in light inten- On average, xylem pressure dropped after a delay of
sity, was 10·9 ± 9·3 mV (n = 19). At the end of the depo- about 2–3 min by 0·14 ± 0·07 MPa (n = 9). This value was
larization phase, the trans-root potential had changed by reached within about 22 min. Hyperpolarization always
10 ± 15 mV (n = 15) compared to the value recorded at occurred immediately after the light was switched on.
low light intensity. Consistent with wheat, hyperpolarization preceded the
light-induced decrease in xylem pressure by about
4 2–3 min. In other experiments than the one shown in
Preliminary experiments performed on very young, transpiring
plants (7–14 d old) showed that only depolarization occurred upon
Fig. 8b, the hyperpolarization response was often less
illumination (data not shown). This shows that variations of the pronounced; the mean value was 8·1 ± 7·4 mV (n = 10).
light-induced changes of the trans-root potential may depend on Subsequent depolarization was only observed in half of the
age. experiments.
© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
862 L. H. Wegner and U. Zimmermann

Bringing back the light intensity to a low level down to a pressure of about – 0·3 MPa; Fig. 6a,b), (2) erro-
(10 µmol m–2 s–1) frequently resulted in an instantaneous, neous potential readings due to tip clogging can easily be
rapid depolarization followed by a slow hyperpolarization. identified and eliminated by pressure pulses generated
In analogy to wheat, the hyperpolarization phase was not within the probe by displacement of the metal rod and (3)
always observed. Xylem pressure responded after 1–3 min complications arising from cell damage and/or incomplete
and shifted to more positive values; within 60 min, a new sealing around the inserted microcapillary can apparently
steady state was established. be ignored because of the magnitude of the radial root
The observed delay in the response of xylem pressure resistances, the membrane potential measurements of
(but not of the trans-root potential) in the root of wheat cortical cells (closely located to the insertion site) and
and maize upon illumination did not result from a delay in because of the high radial reflection coefficients measured
the propagation of the pressure signal from the leaves to at elevated transpiration rates (Schneider et al. 1997a,b).
the roots. This could be demonstrated by simultaneous Compared to the exudate and conventional microelec-
insertion of a pressure probe into the xylem of the middle trode methods employed by other authors to measure the
vein of the leaves and another probe into the xylem of the trans-root potential (see Introduction), the xylem pressure-
roots. A typical experiment performed on maize is shown potential probe has the additional advantage that the
in Fig. 9. After probing of a leaf vessel (arrow ‘A’) a sec- hydraulic and electrical responses in the xylem conduit
ond probe could be introduced 2 min later into a vessel in upon changes of environmental factors can be measured
the root (arrow ‘B’). Both probes showed nearly the same simultaneously.
pressure recordings after insertion at a light intensity of The present study provided us with the first results on
10 µmol m–2 s–1, except that the value of the pressure in the changes of the xylem pressure and associated changes of
xylem in the root was always by about 0·02 MPa more the trans-root potential upon osmotic stress and illumina-
positive than that in the leaves. When light intensity was tion, respectively. It is obvious that the response pattern of
increased to 250 µmol m–2 s–1 (arrow ‘C’), both probes the trans-root potential in relation to the changes in xylem
read a drop in pressure, but only after about 2 min. This pressure exceeded simple electrokinetic phenomena
demonstrated that there was no delay in the transmission of (Fensom 1957, 1962; Fensom & Dainty 1963). The
pressure changes from the leaf to the root. Interestingly, the osmotic experiments performed on maize roots gave some
pressure gradient between the vessels in the root and leaves indications that electrokinetic effects played a role because
increased to about 0·4 MPa once a steady value had been the changes in the trans-root potential were associated with
reached after the ‘overshoot’ response. changes in the xylem pressure. However, it can be argued
that these changes did not reflect the occurrence of stream-
ing potentials, but rather changes in the membrane poten-
DISCUSSION
tials of root cells generated by osmotically induced
The results presented here demonstrate that the xylem changes of the turgor pressure of these cells. Turgor pres-
pressure-potential probe yields precise data for the trans- sure-dependent transport processes in the plasmalemma
root potential. The main arguments for this conclusion are: membrane of algae and higher plants are well known
(1) the xylem pressure recordings indicate that the poten- (Zimmermann & Steudle 1974; Zimmermann & Beckers
tial and xylem pressure reading probe is definitely placed 1977; Wendler, Zimmermann & Bentrup 1983; Stahlberg
in a vessel without rupture of the water column (at least & Cosgrove 1997; Lew 1996).
In any case, the response of the trans-root potential to
illumination in both species and to xylem pressure oscilla-
tions in wheat suggests that the changes in the trans-root
potential were not directly or exclusively coupled to
changes of pressure-induced volume flow. This view is
supported by the finding that upon illumination hyperpo-
larization preceded changes in xylem pressure both in the
leaf and in the root (see Figs 8 & 9). In addition, the inves-
tigations of the oscillation phenomena in wheat have
shown that a significant depolarization of the trans-root
potential was only observed once a maximum xylem
tension had been established (Fig. 6). Consistently, hyper-
polarization was generally (but not always, see Fig. 6b)
most pronounced at a minimum of xylem tension. The
Figure 9. Simultaneous probing of a vessel in the leaf and in the more or less coincidence of the extremes in tension with
root of a 17-d-old maize seedling. As indicated by the arrow ‘A’, a the de- or hyperpolarization response of the trans-root
xylem vessel in the midrib of the second leaf could be probed at
time zero. About 2 min later (arrow ‘B’), a second probe was
potential was particularly distinct when both xylem pres-
inserted into a xylem vessel in the root about 20 cm above the tip sure and trans-root potential oscillated very regularly.
(total root length 27·6 cm). At arrow ‘C’, light intensity was While these results provide evidence for a regulatory
increased from 10 to 250 µmol m–2 s–1. link between (the steady state value and magnitude of) the
© 1998 Blackwell Science Ltd, Plant, Cell and Environment, 21, 849–865
 Xylem pressure and trans-root potential 863

xylem tension and the electrical response, the data are not funded by the Deutsche Forschungsgemeinschaft (Zi
sufficient to identify the origin of the trans-root potential 99/10–1) to U.Z.
changes. However, the finding that in wheat the oscilla-
tions of xylem pressure co-oscillated with the trans-root
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