Biophysical Chemistry, 33 (1989) 161-176 161

Elsevier

BPC 01352

Growth and electric current loops in plants

Kiyoshi Toko a, Takanori Fujiyoshi ‘, Chikako Tanaka b, Satoru Iiyama b,

Toshirou Yoshida a, Kenshi Hayashi a and Kaoru Yamafuji a
’ Department of Electronics, Faculty of Engineering. Kyushu University 36, Fukuoka 812
and b Department of Home Economics, Women’s Junior College of Kinki University, Iizuka 820, Japan

Received 31 March 1988

Revised manuscript received 12 January 1989
Accepted 19 January 1989

Multicellular system; Electric current; Proton accumulation; Growth; (Plant)

A theory is presented for a relationship between ion accumulation and electric current loops in multicellular systems such as the roots
and stems of higher plants. A network of electric circuits shows that the electric current transported across the cell membrane flows
between an elongating region and a mature region, not only in roots but also in stems. In roots, ions constituting the extracellular
electric current flow in the external aqueous medium, while in stems an electric current of comparable density flows within the
epidermal cell wall. Based on this theoretical result, electric isolation between the elongating and mature regions was made in the case
of both roots and stems. The speed of growth during the initial stage was greatly decreased due to a change in the distribution of
protons around the surfaces of the plaut by cutting off the electric current loop. Electrochemical calculation shows that ions are not
always accumulated at the efflux site, since the ion distribution is strongly affected by the relation of the magnitudes between the
electric field and electric current. The results calculated for the electric potential and pH distributions around the root agree with
experimental data.

1. Introduction multicellular systems [5,6]. In roots of barley and

bean, electric spatial patterns have also been shown
Spatial patterns of electric potential and elec- to appear with growth [7-91. While these experi-
tric current often appear as macroscopic order mental studies suggest that such patterns play a
during the processes of growth and regeneration in role in growth, differentiation and morphogenesis
both plants and animals. The appearance of an [lO,ll], the detailed mechanism including sponta-
electric current pattern precedes rhizoid formation neous stabilization of electric spatial patterns has
in the brown alga Fuctu [l]. Characean species remained unelucidated.
develop alternating bands of acid and alkaline A possible approach to the clarification of the
regions along their cell walls under illumination, mechanism underlying the appearance of electric
and localized elongation occurs in acid regions. patterns is to employ nonequilibrium tbermody-
The electric current flows from the acid to alkaline namic theory taking into account the electrochem-
regions [2-41. It has long been known that an istry of the biological systems concerned. It has
inhomogeneous electric pattern appears along the been shown that, for the appearance of morpho-
surface of the onion root, a typical example of genetic polarity in Fucus eggs, the electric spatial
pattern associated with the polarity is a self-
Correspondence address: K. Toko, Department of Electronics,
organized structure arising under far-from-equi-
Faculty of Engineering, Kyushu University 36, Fukuoka 812, librium conditions [12,13]. Furthermore, the band
Japan. structure in Characean species is realized cooper-
0301~4622/89/$03.50 0 1989 Elsevier Science Publishers B.V. (Biomedical Division)
162 K. Toko et al./Heciric current loops in plants

atively through simultaneous activation and in- pointed out from considerations based on electro-
activation of molecules of the H+ pump within the chemistry that ions are not always accumulated
plasmalemma along the cell [2,3,14]. Such a band under given conditions but, rather, are depleted at
structure can be interpreted as being of the self- the efflux site, from which the ions are pumped
organized [15], i.e., dissipative type [16]. The out. Accumulation of protons is shown to depend
spontaneous emergence of an electric pattern oc- on the relation of the magnitudes between the
curs through a self-organized process of ion flux electric field and electric current. The results
across the membrane. calculated for the pH and electric potential agree
In the present paper, we investigate the mecha- with the experimental data demonstrating acidifi-
nism of action on the growth process by the cation near the root tip, and furthermore, provide
electric current formed through the above self- an explanation for the possibility of alkalinization
organized process. For this purpose an electric [7] near the elongation zone of roots.
circuit network is useful for expressing the electro-
physiological characteristics of a system compris-
ing an elongation and a mature zone. Particularly 2. Basic experimental data on electric potential and
in plants, the loosening of cell walls via protons is PH
known to be a controlling factor during elongation
growth [17,18]. In discussing the growth of plants, In this section, we describe briefly the basic
therefore, it is necessary to investigate proton ac- data on roots that are necessary for the theoretical
cumulation near the cell wall caused by the elec- description. The experimental procedures and re-
tric current, which is produced by activated H’ sults are detailed in our previous reports [8,9,19].
pumps. However, the connection between the pH As for stems, the fundamental data on the electric
and electric current is obscure, since the pH is low potential will be given explicitly in section 3.
in regions of high electric potential near the surface Seeds of a germinating hypogeal plant (azuki
of Chiiracean cells [3], whereas the root has a low bean; P. chrysanthos) were allowed to soak for 3
pH at the root tip of low surface potential (fig. 1) h, and were then sown on moistened filter papers
Fu91. in darkness at 30 f 1” C. Roots emerged 1 or 2
Thus, the spatial electric pattern in roots and days later. Roots longer than about 5 cm were
stems of higher plants was first investigated using used.
a model of the electric circuit network. The ex- The electric potential near the surface along the
istence of electric current loops connecting the root was measured by the use of a multi-electrode
mature region to the elongation zone was demon- system [9,19,20]. About 30 pipette electrodes were
strated not only for the root but also for the stem. arranged at 1.5-mm intervals along the root. The
Whereas the electric current within a conductive electrode tip was located 0.5-1.0 mm from the
medium was determined for roots [7], it could not root surface, care being taken to ensure that direct,
be measured for stems, which usually do not grow mechanical contact with the root surface was
in conductive medium but in air. Probably for this avoided. The reference electrode was placed at a
reason, the suggestion that longitudinal flow of distance from the root. The whole apparatus is
ions within tde epidermal cell &all affects the composed of electrodes as electric sensors, a
growth has not been proposed for stems. The hand-made circuit for sampling data adequately
present theory shows that in stems a considerable and a personal computer for processing and
amount of electric current can flow longitudinally arranging the data. One scan over all the elec-
in the epiderrnal cell wall. trodes can be performed within 3 s.
Secondly, experiments were performed in order The pH near the root surface was measured
to’ demonstrate that interruption of the electric with a pH electrode (Iwaki Glass, IW202), the tip
current loop markedly affects the speed of growth diameter of which was less than 1 mm.
in both roots and stems of higher plants (Phaseo- Fig. 1 shows the surface electric potential,
lus chrysanthos, Vigna unguiculata). Thirdly, it is surface pH and growth speed along the root in a
 K. Toko et d/Electric current loopsm planis 163

ton pumps work in opposite directions, i.e., back-

to-back. Fig. 2 illustrates this situation, which may
hold for both the root [23] and stem (211. In this
figure, the symbols x, p and s denote the xylem,
parenchyma and surface, respectively. The mea-
sured electric potential is represented by I? For
example VPSdesignates the potential at p measured
from s. The e.m.f. and electric resistance of the
6
s -4

-6
v
t
Distance from
tip (mm)
root electrogenic pump are given by E, and r,, respec-
tively. The e.m.f. of the passive (diffusive) channel
is Ed, the resistance being r,. The additional sub-
scripts 1 and 2 concern the quantities at the
Fig. 1. Properties near the root tip. ( -) Electric potential, plasmalemma of epidermal and cortical cells re-
(o-- ----•) pH and (bars) growth speed. A root of approx. 8 sponsible for the generation of VPS and those at
cm in length was placed in 0.1 mM KC1 solution at 30f 1°C. the symplast/xylem interface producing VPX, re-
Growth speed is averaged over 32 roots longer than 5 cm. The
spectively. The parallel resistance due to the
elongaktg region exists between 1 and 5 rum from tip, where
the surface electric pot&al and pH are relatively low. Casparian strip, which is an insulating structure
against ion and water movement, is so large that it
has been omitted as it effectively amounts to
narrow region 12.5 mm from the tip. The intact infinity. The parenchyma can be regarded as a
root was placed horizontally in the chamber with cytoplasrnic continuum [24-271.
0.1 mM KC1 solution. One can see that elongation Although ions can easily flow in the external
is limited to l-5 mm from the root tip while a aqueous medium in the case of roots, there is no
peak of electric potential exists near 6 mm from conductive medium in the case of stems, which
the tip. The mature region extends mostly behind
approx. 6 mm from the tip. While fig. 1 shows
acidification at the root tip and in the elongation
zone, alkalinization relative to the elongation zone
was sometimes observed close to the root tip. The
surface electric potential was low on the side of
the tip comprising the elongation zone. On the
whole, the pH was high for high surface electric
potential under these experimental conditions. The
patterns of pH and electric potential disappeared
under anoxia [20].

3. Electric circuit network

3.1. Electrophysiological model around the elongat-

i XYLEM1SYMPLAST I.
SURFACE

ing region for roots and stems

Fig. 2. Electrophysiological structure in the radial direction of
the root and stem. CS, Casparian strip. This model is il-
There are two independent electromotive forces lustrated according to previous electrophysiological studies
(e.m.f.) in the radial direction of the electrophysio- (21-231. Subscripts s, p and x to Y designate the surface,
logical structure in the stem as well as the root parenchyma and xylem, respectively. The electrogenic pump is
described by the electromotive force & and the electric resis-
[21-231. One is located at the surface of the tance r,, the diffusion channel being expressed by Ed and ra.
parenchyma symplast, the other between the Subscripts 1 and 2 refer to the surface and xylem sides,
parenchyma and xylem. These iwo kinds of pro- respectively.
164 K. Toko ei al. /Electric current loops in plants

mature (m) regions. In this model Ei and 5 for

i = 1, 2 are expressed as:

with the terms Eei, etc., defined in fig. 2. The

longitudinal resistance of the symplast between
the tip and elongating regions is denoted by RF),
that between the elongating and mature regions
being represented by R, (em). The longitudinal resis-
tances at the surface are given by Rf@ and Rfm).
X P s
Interpretation of R, requires some caution: In the
Fig. 3. Electric circuit network model of electrophysiological case of roots, R, is mainly composed of the
structures of roots and stems. Tip, elongating and mature resistivity of the surrounding conductive medium.
regions are indicated by t, e and m, respectively. In the radial
direction, the electromotive force is expressed by ,JZ with
For stems, on the other hand, R, should express
electric resistance r. These are expressed by the electric ele- the longitudinal resistanci of the apoplast, i.e., the
ments defined in fig. 2. Subscripts denote the radial positions: epidermal cell wall. Since the electric potential is
s, surface; p, parenchyma; x, xylem; 1, surface side; 2, xylem homogeneous along and within the xylem [30], the
side. Superscripts denote parameters with respect to the longi- short circuit is drawn at the xylem in fig. 3. The
tudinal direction. Note that the electric resistances between the
electric current on the longitudinal branch be-
tip and elongating regions are introduced by Rb? and RF),
together with those between the elongating and mature regions, tween the elongating and mature regions at the
R’p”“) and R,(emJ. The surface refers to the aqueous medium in surface is denoted by i,, and that between the tip
the case of roots, and the cell wall of epidermal cells for stems. and elongating regions by i,. The radial electric
Electric currents flowing in the branch are defined by i,, i,, i,,
(0, i(C) and i(m).
currents across the plasmalemma of the epidermal
i4, t
cell in the tip, elongation and mature regions are
designated i(‘), ice) and icm’, respectively.

3.2. Electric current loops formed in radial and

longitudinal directions
usually grow in air. However, we can expect the
presence of a longitudinal ionic flow even for The electric current flowing in each branch
stems. Ions are able to diffuse not only across the defined in fig. 3 is explicitly given by
parenchyma symplast but also in the cell wall of i, = (K(m) - <‘“‘),/RQ”m’,
the epidermis. In fact, the Characean cell can also
develop the acid/alkali pattern when it is sus- i, = ( v/e1 _ Vs(t+‘Rrte),
pended in air [28,29]. Here, the electric current
i, = i, + (V;‘) - qt)),‘R’d’),
can be considered to circulate longitudinally within
the cell wall and the protoplasm between the acid i, = i, + (rim’ - Vi” )/Rfm). (2)
and the alkaline regions, although of small magni-
tude in the cell wall due to the low conductivity.
Even in the case of stems, therefore, an electric 3.2.1. Roots
current can be formed within the epidermal cell We first calculate the electric current in the
wall. We must take into account the longitudinal case of roots. For a root of typical size, we adopt
flow, which is not considered in fig. 2. Lcte) =i 2.5 mm as the average distance between the
Fig. 3 illustrates the electric circuit network, root tip and elongation zone, L@“‘)= 8.0 mm as
improved so as to express the longitudinal electric the distance between the elongation and mature
connection among the tip (t), elongating (e) and zones, and radius r = 0.5 mm for the symplast (fig.
 K. Toko et al./ Electric cwwnf loops in plants 165

The electric current densities in the external con-

ductive medium amount to 0.19 PA/cm2 between
the elongating and mature regions, and 0.10
PA/cm2 between the tip and elongating regions.
i(T) These values are consistent with current densities

i(D
T i2
measured by using a vibrating probe [7,34,35].
From the flux continuity, we can also estimate the
radial flow across the surface i(l), i@) and i@“)
il
defined in figs. 3 and 4. These parameters are
t
related to i, and i, in terms of the current den-
i(M)
sity:

i(‘) = -i,/2lrrL(‘), ice) = (i, - i1)/27rrLce),

(4)
icm) = i,/212rLcm’.

Fig. 4. Definitions of electric currents and lengths. The lengths We obtain i(‘) = -0.48 PA/cm2 (< 0), i(“‘=
of the tip, elongating and mature regions are denoted by L(‘), -0.10 PA/cm’ (-G 0) and itm)= 0.26 PA/cm2
L@) and LCm),respectively. Distances between the two regions
(> 0). These values are of comparable order to the
arc LCtc)and LCern)with the radius denoted by r. For roots,
L(‘) = 1.25 mm, L(‘) = 5.25 mm and L”“’ = 4 mm; for stems longitudinal flux. Fig. 5a illustrates the resultant
L(‘) = 6 mm, L(” = 18.5 mm and L(“‘) = 12.5 mm. flow pattern within a region approx. 1 cm in
length comprising the tip, elongating and mature
regions. Electric current loops are formed in the
4). From fig. 1, the surface electric potentials may area including the tip, elongating and mature re-
be estimated to be cls(‘)= - 5 mV, V/j = - 4 mV gions.
and yStrn)= 2 mV. The electric potentials in the We feel that some comments are required con-
parenchyma may be evaluated as V$) = - 95 mV, cerning this result. The surface electric potential
k’@)= - 105 mV and L’(m)= - 75 mV [31]. As for near the root tip is so dynamic that it exhibits
thPe electric resistance ii the longitudinal direction large negative values or sometimes relatively posi-
of the parenchyma, we assume the resistivity to be tive values. This tendency is evident for roots of
2.5 x lo’&? cm [32,33]. Using this value, RF) and shorter length [5,19], the electric current of which
R’“’ can be estimated as 7.94 x lo3 and 2.55 X is measured via a vibrating probe [7,34,35]. There-
1J4 52, respectively. The resistivity of the external fore, the sign and magnitude of i2 readily undergo
aqueous medium, e.g., 0.1 mM KC1 solution, is change whereas those of i, do not. This means
approx. 4 X 10’ D cm. In analogy with the Char- that both i@) and i @) display considerable varia-
acean cell [14,15], electric current flows within the tions while icrn) does not. If I/S@)equals -8 mV,
cylindrical region of the conductive medium,
D
whose radius is at most 5-times greater than that b

of the root. The longitudinal resistances Rf@ and

R’,e”‘) are therefore estimated to be 5.30 x lo4 and
1.70 X 10’ f2, respectively.
Substitution of these values into eq. 2 yields the
following:

ir = 3.53 X 1O-8 A (> 0),

il=1.89x10-8A(~0),
(3)
’ = -1.24x10-6A(<0),
13 Fig. 5. Theoretical results on electric current loops for root (a)
and stem (b). The longitudinal direction is illustrated in the
i4=1_21x10-6A(>0). same way as in figs. 3 and 4.
166 K. Toko et al./ Electric current loops in plants

for example, then i2 amounts to 7.55 x 10s8 A, The electric current densities can be evaluated
which leads to i(*)= -1.92 PA/cm2 (< 0) and as i, = 1.8 PA/cm2 (r 0) and i, = -1.5 PA/cm’
i@)= 0.24 PA/cm’ ( r 0). In this case, the electric ( < 0) if the thickness of the epidermal cell wall is
current comprises efflwr from the elongation zone assumed to be 1 pm. It is noticeable that the
as well as from the mature region. Influx occurs above values are comparable to or greater than the
only in the tip region. Since a positive peak always current densities flowing around the root in a
appears around 1 cm, located within the mature conductive medium. The density of electric cur-
region for roots of greater length [8,9,19], the rent traversing the plasmalemma of epidermal and
mature region behaves as a source of electric cortical cells can be calculated from eq. 4, yielding
current for the external conductive medium. the values i(‘) = 2.54 X 10e4 PA/cm2 (> 0), ice) =
- 1.82 x 10e4 PA/cm2 (< 0) and iCm)= 1.46 x
3.2.2. Stems 10e4 PA/cm’ (> 0). These values of the radial
Typical values of the dimensions for a stem are fluxes are considerably smaller than those of the
as follows: L(‘@= 1.2 cm, L’““) = 2.5 cm and r = longitudinal fluxes i, and i,. This occurs because
0.1 cm. The surface electric potential in each the current must flow longitudinally in the narrow
region is l$cr)= -5.8 mV, K@)= - 17.3 mV and apoplasmic space of epidermal cells, while the
I’@‘)= 11.4 mV; the electric potential within the radial current is generated across the relatively
&nplast amounts to PF’ = - 110.7 mV, I$@‘= wide area of the plasmalemma. The resultant flux
-127.2 mV and I/d”‘= -117.2 mV [21]. In the pattern is illustrated in fig. Sb, the above-men-
case of stems, if the value adopted for the resistiv- tioned area being composed of the tip (hook part),
ity of the parenchyma is taken to be the same as elongating and mature regions, reaching approx. 4
that of the root (= 2.5 x lo2 s2 cm), the values of cm as a whole. Two large loops are formed be-
Rb’“’ and Rtm) may be determined as 9.56 x lo3
tween the xylem and the surface around the elon-
and 1.99 X lo4 9, respectively. There is no con- gating region.
ductive medium surrounding a stem, but an elec-
tric current can flow within the cell wall instead.
The longitudinal resistance per unit length of cell 4. Electric isolation
wall has been determined to be approx. 10’ Q/cm
[28,33]. Thus Rr) and R$“) show values of 1.2 X The experimental procedures empIoyed for
10s and 2.5 x 10s Sz, respectively, which are much electric isolation are detailed in appendix A. With
greater than that of the external conductive roots, the elongating region was electrically iso-
mcdium_surrounding the root. lated from the mature region. The results obtained
Eromeq. 2, we obtain can be summarized as follows [19]: roots being
i .:
i, = 1.15 X.IO-r” A (> O), treated showed slower rates of elongation than the
control. The pH displayed lower values in the
i,= -9.58~10~‘lA(<O), mature-region side compared to the elongating-re-
(5)
i3 = -1.73~10-~A(<O), gion side. In contrast, the pH showed lower values
in the elongating region for the control group.
i,=5.03~10-‘A(>O).
The theoretical result described in section 3 is
This result implies that the magnitude of the elec- that an electric current flows longitudinally along
tric current flowing within the cell wall is about a root in the external medium to the tip side. By
0.01% of that flowing in the xylem and also in the taking into account the mechanism of acid elon-
symplast. Efflux occurs from the surface side in gation [17], a straightforward explanation for these
the mature and tip regions; however, electric cur- results may be given as follows: Protons are trans-
rent flows into the parenchyma in the elongating ported from the mature-region side to the tip
region (fig. 5b). Within the xylem current flows region under the usual conditions, and accumulate
downward from the elongating region in the direc- in the tip and elongating regions, thus causing
tion of both the tip and mature regions. wall loosening. Electric isolation inhibited longitu-
 _)
K. Toko et al. /Electric current loops in plants 167

dinal ionic flow in the external conductive medium, Therefore, we can consider the electric current in
resulting in depletion of protons in the elongating the control group to be composed partly of the
region; a decrease in the rate of wall loosening flow of protons between the elongating and ma-
resulted, and consequently elongation was re- ture regions through the aqueous phase contacting
tarded. the surface of the stem. Electric isolation inter-
Fig. 6a concerns electric isolation between the rupts the extracellular current loops. Since. both
elongating and mature regions in the case of stems. control and electric-isolation groups are in contact
The only difference in experimental details be- with wet filter paper, the difference in growth
tween the electric-isolation and control groups is speed is determined by whether or not electric
the difference in shape of the plastic plate, as current loops are formed. The electric loop be-
shown in fig. 10. Stems under electric isolation tween the elongation and mature zones is essen-
grew at a lower rate than the control group. The tial. In fact, the cut-off of electric current flowing
pH in both mature and elongating regions showed between different parts of the mature region did
higher values for stems under electric isolation not affect the growth of stems (fig. 6b).
compared to the control group, although the pH In the present experiment, the electric current
reflects only qualitatively the pH of the cell wall flowing within the aqueous phase contacting the
due to the presence of the cuticle. This result surface of a stem was interrupted directly. This
provides an explanation for the slow rate of elon- electric current must flow within the cpidermal
gation of stems under isolation, since wall loo- cell wall under the normal conditions of being
sening is scarcely able to occur at high pH values. wholly surrounded by air, as depicted in fig. 5b;
./.
!

Time (hr) Time (hr)

Fig 6. Experimental results on electric isolation for the stem. Vertical bars represent standard deviations. (Inset) Tabulation of pH
value in each region (e. elongating; m, mature region). (0) Control, (@) electric isolation, (0) normal conditions, i.e., stems growing in
air free of the attached wet paper. (a) Electric isolation between elongating and mature regions, i.e., at 18 mm from the cotyledonary
node. 25, 27 and 18 stems were used at 30° C for the control, isolation and normal groups, respectively. pH vahtes are. those
measured 2 h following the start of the experiment, where the respective numbers of stems used were l?, 8 and 13. Among the control
eight stems showed almost the same pH values’aa in the normal group. dH in the elongating and mature regions incrcaxd with time
in stems under electric isolation. (b) Isolation between mature regions, i.e., at 30 nim Prom the base at 30’ C. Respective numbers of
8.siemswere19.22 and 20.
 168 K. Toko et al./Electric current loops in plants

this forms an intra- and extracellular electric cur-

rent loop together with the flow in the multicellu- Ji
lar continuum comprising the symplast and xylem. Jk
In both roots and stems, delay in the rate of
growth under electric isolation occurred only dur-
ing the initial stages. Since the control group was
treated in the same way as the group under isola-
tion (figs. 9 and lo), the initial low speed under
isolation is not due to the effect of some mechani-
cal perturbation associated with the experimental Fig. 7. Concentrations and electric potential in a one-dimen-
setup of the plant, but rather results from inter- sional system with coexistiq monovalent cations and anions.
ruption of the electric current. One reason why Subscripts i (=1,2 ,..., m) and k (=1,2 ,..,, n) refer toca-
tions and anions, respectively. Fluxes Ji and Jk occur at the
complete cessation of growth did not occur may in
two boundaries x = 0 and x = L. Electric potential V (or
part be due to incomplete isolation: the surface of electric field E) is determined self-consistently with the distri-
the root or stem always, became moist even when butions of ions.
the moisture was wiped off with filter papers.
Another possibility is that partial efflwr from the
elongating region may occur [19,34-361. Once the
elongating region has changed into the mature simple example of the case of constant E is
region, it becomes a source of electric current, discussed in appendix B.
thus giving rise to the usual growth speed. Concentrations ci and ck in the stationary state
From the viewpoint of acid elongation of the can be solved easily [37], as shown in appendix C:
cell wall, the behavior of protons may be im-
portant. Section 5 addresses the question of the c.1 = &C(X)+A&,_ $E][(z-gy-=‘s
electrochemistry for the relationship between ion I

accumulation and electric field.

_ ( E+ ~)l-=‘s]-lc(x)-T/s,(74
5. Ion accumulation and electric field
c, = - -&C(X) k

A one-dimensional system composed of an l+T/S

I+T/s
-1
aqueous solution of 0 <x < L containing some

( * 1c(xy,
ion species is considered here. At the two
boundaries, transport of cations (A) and of anions
SL
(Jk) takes place. The situation is demonstrated in - z-
0)
fig. 7. The ion species are limited to monovalent
ions. The flutes ji for cations (i = 1, 2,. . . , m) and
jk for anions (k = 1, 2,. . . , n) in the medium are
where c(x) Ii IS )

given by

explicitly given by c(x)=P+-s5

2P(2 i
) (8)
ji = -Di ac& + uiEci,
with Z being defined as:
_ik = - Ok aC,/aX - ukECk, (6)

c=_ 1 Lc(x)dn= &= i Ck. (9)

where c, and c, designate concentrations, Di and /
LCI i-l k-l
Dk diffusion constants, ui and uk electric mobili-
ties and E the electric field in the medium. Sub- The quantities C, A, T and S are constants
scripts i and k refer to cations and anions, respec expressed by J, and Jk; their forms are defined by
tively. The electric field E is dependent on n. A eqs. C4 and C5. The parameter B is defined by eq.
 K. Toko et al./ Electric current loops in plants 169

C2, and is proportional to the absolute tempera- s) for K+ and J, = 2.5 X lo-3 PA/cm* (= 0.025
ture, being approx. 25 mV at-room temperatures. pmol/cm’ per s) for Ht. The value adopted for
The electric field E is given by proton flux is of the same order as that estimated
from fig. 1 using the discrete form of eq. 6:
E(x) = T/2c(x). (10)
J, = u2[H+][ -p API-I t Al’]/l, (13)
The potential difference between x = 0 and an
arbitrary position x is calculated as where u2 represents the electric mobility of pro-
tons (= 3.626 X lop3 cm*/V per s), [H’] proton
V(x) = -i%(x) dx concentration at the midpoint between the tip and

1 (11)
mature regions, i.e., at around 3 mm from the tip,
z+S(L/2--x)/2fi and I (= 6 mm) the distance between these re-
=- PT
sh [ z+ SL/4/9 * gions. ApH ( = 0.2 pH units) and Al/’ (= 9 mV)
denote the respective differences in pH and elcc-
If anion transport does not occur at the tric potentials between the two regions.
boundaries or anion flux is negligible compared The calculated results are plotted in fig. 8. The
with cation flux, eqs. 7a and 7b become pH profile and electric-potential pattern from 1 to
6 mm agree well with the experimental data in fig.
1: The H+ concentration is greater and the electric
potential lower in the elongating and tip regions
compared to the mature region. This tendency is
(124 the same as that of case (ii) for J < UEC, discussed
in appendix B (see fig. 11). In fact, this inequality
Ct = &$(x)/z. (12b) can be easily confirmed by putting J = Jz, u = u2,
Eqs. 7,10,11 and 12 are the expressions for the E = AV/l and substituting the value of the proton
concentrations and electric field (or electric poten- concentration in the mature region (= 10m5.* M,
tial) in a one-dimensional system where cations the value at x = 0) for cO. The present case con-
and anions are transported across the two cerns a strong electric field, since the term com-
boundaries. In roots, ions are transported across prising E is larger than the flw J. Since the IL+
the membrane to flow in the aqueous medium. We concentration is greater at the efflux site of high
consider the membranes in the mature and tip-side electric potential, the electric field can be con-
regions (including the elongating region) as the sidered as being generated mainly by the K+ flux.
boundaries at x = 0 and L, respectively (fig. 7). Protons are accumulated near the elongating re-
The aqueous medium surrounding the root corre- gion due to the resultant electric field. The anions,
sponds to the region between x = 0 and L. Cl- and HCO;, are distributed so as to satisfy the
As major ion species flowing around the root, requirement of electric neutrality.
we deal with H+, IL+, HCO; and Cl- by taking The behavior of ci for H+ can be easily under-
account of the experiment of fig. 1. We assume for stood from eq. 12a. The coefficients for the first
simplicity that two kinds of cations, H+ and K+, and second terms are positive. The first term
are mainly transported across the membrane, since. demonstrates that ci is proportional to c(x), which
the rate of anion transport is low [38,39]. From the is nearly equal to the K+ concentration. The sec-
theoretical result (fig. 5a) and the experiment of ond term signifies that the trend in ci is opposite
electric isolation [19], electric current flows from to that of c(x). Since Z and 4 are small, the
the mature to tip-side region. In the present calcu- second term predominates. Thus, the behavior of
lation, the current is composed of H+ and K’ and ci is the reverse of that shown by c(x), i.e., Ht is
amounts to approx. 0.2 PA/cm’ as evaluated accumulated in the region where K+ is depleted.
from eq. 3 using the electric circuit network. We Increasing KC1 concentration reduces the elec-
assume that the largest contribution is that due to tric potential. In this case, the coefficient for the
K+, i.e., J1 = 0.25 PA/cm’ (= 2.5 pmol/c& per second term in eq. 12a is of negative sign; it
170 K. Toko et al./ Electric current loops in plants

ship between the electric current and the accumu-

lation of protons around the elongation zone on
the basis of the acid-growth mechanism [17].
(3) Electrochemical analysis shows that the ac-
cumulation of ions is strongly affected by the
magnitude of the electric field. Protons are not
always accumulated at the efflwr site. Theoretical
calculation reproduces well the profiles for pH
and electric potential in fig. 1. Protons accumulate
-6
Distance from at the efflux site when the flux is large in relatively
roof tip (mm)
high concentrations of coexisting ions, thus ex-
Fig, 8. ‘Theoretical results on electric potential, pH and K+ plaining the observed data [3,7,35,41].
concentration near the tip of the root. ( -) Electric poten-
It has been reported that the value of the pH in
tial, (------) pH and (.-.-.) K+ concentration near the
surface, in 0.1 mM KCI solution. (* - - . . -) pH in 1 mM KC1
the xylem is lowest around the elongating region
solution. Average pH was set to 5.8. Note that H+ is accu- of stems [21]. This result can be explained theoret-
mulated in the elong&ng region while the,K+ concentration is ically. Fig. 5b provides confirmation that the elee-
high in the mature region. Positions 1 and 6 nun from the root tric current within the xylem flows downward to
tip correspond to x = 2, ( = 5 nun) and x = 0 in fii. 7, respec-
both the tip and mature regions from the elongat-
tively. Electric potent&l at x - 0 is taken as equal to 4.0 mV.
H+ and K* are assumed to be extruded from the x = 0 site, ing region. The elongating region is that part
i.e., the mature region. The results agree well with fig. 1. which displays efflux within the xylem. The elec-
tric-potential difference is known to be zero within
the xylem [30], and hence the electric field is zero.
follows that ci for H” shows behavior similar to This means that the present case falls within the
that of c(x), which is almost equal to the K+ category of weak electric field, i.e., case (i), shown
concentration. Calculation with 1.0 mM KCl, in fig. 11. In this case ions must accumulate
therefore, demonstrated that alkalinization oc- around the site of efflux. Thus, the abovamen-
curred in the elongating region, where the electric tioned occurrence of the lowest pH values within
current comprises influx, as shown by the dotted the xylem around the elongating region is reason-
line in fig. 8. This provides an explanation for the able.
alkali&ation observed previously [7,40]. Protons In the present experiment, the cuticle of the
are accumulated at the efflux site under these stem was not abraded. Many studies have shown
conditions, as observed previously in roots and that abrasion is necessary for investigation of the
Characean cells [3,35,41]. quantitative effect of acidification on growth
[42-451. However, ions including protons cannot
always pass through the cuticle [46-511. The cuticle
6. Discussion does not behave as an insulator but as a very weak
conductor. On a time scale of several tens of
The following points have been demonstrated minutes, protons can move into and out of ,a tissue
in the present paper: across the cuticle [SO].Since elongation was moni-
(1) Theoretical analysis using an electric circuit tored over a few hours (fig. 6), protons might pass
network suggests the, existence of electric current through the cuticle. Slight alkalinization on elec-
loops formed around the elongating region in both tric isolation (fig. 6a) may reflect a qualitative pH
roots and stems. While the currem usually flows in change in the cell wall resulting from buffering
the external conductive medium for roots, it oc- activity of the cell wall [52] and the presence of
curs within the epiderrnal cellwall of stems. the cuticle. In addition, the experimental setup
(2) Interruption of the surrounding electric cur- might have a favorable effect on”the electric con-
rent results in the retardation of growth in both nection between the cell wall and medium due to
the root and stem. This implies a causal relation- the moist filter paper contacting the stem. A simi-
,
 K. Toko et aI./ Electric current loops in plants 171

lar method was also employed in a previous study ment altered the electric potential difference be-
of the effects of acidification and the geotropic tween the parenchyma and organ surface [53].
response of hypocotyls [46]; acidification was This experiment does not appear to show a con-
found to exert a remarkable effect. Furthermore, nection between the electric current loops and
the effect of intentionally applied acid was not growth. However, we are certain of the participa-
investigated in the present study; however, the tion of electric current loops in growth under the
effect of a spontaneously formed electric current usual conditions, i.e., in the absence of externally
was studied using electric isolation. In the control applied stimuli, where growth occurs in the
group, one may consider that protons can flow customary manner. If the stem is subjected to a
according to the electric current even in the pres- number of stimuli, an overall change, such as a
ence of the cuticle; electric isolation disturbs this membrane potential, might appear. In this case,
normal process, resulting in the retardation of proton accumulation occurs firstly at the site of
growth. activated pumps, which leads to enhanced elon-
Further comment is warranted as regards the gation, and then the ions undergo redistribution
difference between the normal and electric-isola- finally, according to the spatial distribution of
tion groups, which showed the smaller elongation pump activation. Situations invoIving such exten-
(fig. 6a). This result appears somewhat unchar- sive activation due to auxin aerosol may corre-
acteristic, since the electric current flows within spond to the weak-field (i.e., strong flux) case,
the epidermal celI wall in both cases. A key to where protons are directly accumulated at the
resolving this apparent contradiction is provided “efflux site, resulting in expansion of the wall, as
by the observations made when Vaseline was ap- has been observed previously [53].
plied around the surface of the stem at a hole From the viewpoint of the acid-growth mecha-
while using a plastic shield (fig. 10). This treat- nism [17], growth retardation of stems under eleo-
ment may cause slight penetration by Vaseline of tric isolation (fig. 6a) can be interpreted as being
the cell wall and/or destruction of the moist layer due to change in pH within the cell wall by cutting
at the surface of the material, which is always wet. off of the electric current between the elongating
As a consequence, increasing hindrance to ion and mature regions. Another possibility may,
movement occurs along the organ’s surface. A however, be considered: the activity of Hf pumps
preliminary measurement showed that the electric is altered due to electric isolation, resulting in the
resistance along the stem increased by approx. 3% cell-wall pH change. This process may be possible
when Vaseline was spread over the surface at the because the ion distribution, which may affect
position separating the elongating from mature activation of the pump, varies according to the
region. Since the resistance of the symplast (c- electric current. Whereas we are unable to make a
R(pCm))may not be altered by this treatment, the definite decision as to which of the two possibili-
small increase in resistance arises from the rise in ties reflects the true situation, we are able to state
that of the organ’s surface (= Rf""B-R~m))..Thu~, unequivocally that the longitudinal electric current
it can be suggested that the longitudinal electric constituting the electric loops is necessary for nor-
current at the surface is decreased by treatment mal growth.
with Vaseline during electric isolation. This leads With elongation of the root, a new electric
to less elongation in the electric-isolation group as pattern is formed sequentially [19,36]. This process
compared to controls. Since the control group is closely analogous to a mechanism of spacing
underwent the same treatment, the difference in patterns suggested previously, based on the pro-
elongation between control and electric-isolation gress zone [54]. Thus, coupling between elongation
groups depends on whether or not an electric and formation of an electric pattern represents a
current loop is formed, connecting the mature and very interesting topic for discussion.
elongating regions through the adjacent aqueous The properties of the electric potential near the
phase, as already mentioned. root tip are rather dynamic. A small peak or a
Application of auxin aerosol to the stem seg- plateau in the electric potential may be observed
112 K. Toko er al,/ Electric current loops in plants

occasionally in the elongating region during growth

[19,36]. Spontaneous oscillations are sometimes
superimposed on the electric pattern [9]. The be-
havior of the electric oscillations varies according
to the position even over the length of 5 mm
limited to the elongation zone (fig. 24 in ref. 19).
These results show that the electric properties are
not necessarily homogeneous within the elongat-
ing region_ Furthermore, the electric pattern for
young roots readily undergoes change. Protons
may be extruded from or sometimes penetrate into
Fig. 9. Experimental setup of electric isolation for roots of P.
part of the elongating region. In addition, with
chrysan~h~ (a) and stems of K unguiculata (b). The root is
respect to geotropism, the ele&ric-current pattern placed horizontally with the stem being oriented vertically.
is drastically changed around the meristem {34].
We can therefore imagine that the electric current
loops are dynamically constructed during growth therefore, it may be desirable to carry out electric
or form in response to external stimuli. isolation by circumferential deletion of the cell
wall at the surface between the elongating and
mature regions. However, this treatment in most
Appendix A cases, damaged the hypoGoty1 organism mechani-
cally. Instead, we attached part of the surface of
Seeds of a germinating hypogeal plant (azuki the hypocotyl to a filter paper moistened with a
bean, P. chlysanrhos) and an epigeal plant (mi- very small amount of aqueous solution, as shown
tori-sasage, I’. unguic~&ta) were allowed to soak in fig. 9b. The chamber was oriented vertically in
for 3 h, subsequently being sown on filter papers order that the stem could grow upwards, as occurs
with 0.01 mM (or 0.1 mM in fig. 1 for electric under normal conditions. In the previous work
measurements in roots) KC1 solution in darkness [36], the system was laid horizontally, a tendency
at 30 f 1’C. of elongation similar to that shown in fig. 6 being
The pH at the surface of the hypocotyl was thus obtained. A large part of the surface of the
measured by attaching a drop of water to the stem is exposed to air. Under such conditions,
surface and then placing a pH electrode of diame- therefore, the electric current can flow within the
ter 1 mm (Fuji, SE17OOGC) at this position for the epidermal cell wall, and furthermore, escape into
horizontally oriented stem. the attached aqueous phase. Similarly to the root,
Electric isolation in the longitudinal direction isolation was accomplished by means of a plastic
of the root was described in detail previously [19]. shield, with Vaseline being applied around the
A chamber with one thin wall of plastic sheet was stem in the hole to isolate the two divisions electri-
utilized for separating the elongation and mature cally. The experimental condition of the stem being
regions, as shown in fig. 9a. The 0.01 mM KC1 in partial contact with a tiny amount of aqueous
solution was poured into the chamber so that the
root could be submerged. In the control experi-
ment, only a partial mechanical shield was con-
structed in order to allow electric connection be-
tween the two divided regions (fig. 10).
In the ease of stems, on the other hand, the
hypocotyl elongates in air under normal condi- Fig. 10. Shield plates for electric isolation (a) and control (b).
Only a partial shield was used for the control so that electric
tions, i.e., not in an aqueous environment. As current might flow &tween the two divisions within the aque-
shown theoretically, the electric current flows in ous medium. Vaseline was applied around the hole, into which
the epidermal cell wall. As an ideal experiment, the root or the stem was inserted.
 K. Toko et al. / Electric current loops in plants 173

phase appears to differ somewhat from the normal

conditions of growth in air. The stem of the con-
trol group, however, exhibited normal growth at
almost the same rate of elongation as that of a
normal stem (see fig. 6). This means that the
present conditions do not represent a close reflec-
tion of those pertaining in physiological situations.

0 L x
Appendix B Fig. 11. Concentration and electric potential in ,the one-dimen-
sional system with a single ion species. Ion concentration is
For roots, the proton concentration is high in denoted by c and spatial coordinate by x. Electric field E is
0.1 mM aqueous KC1 solution near the elongating assumed to be constant. Electric potential V changes linearly
region, where the surface electric potential has a with distance. Flux J occurs at the two boundaries x = 0 and
x = L. Cases (i) and (ii) correspond to the weak- and strong-
low value, as shown in fig. 1. In Characean cells,
electric-field cases, respectively.
on the other hand, protons accumulate near the
cell surface where the value of the electric poten-
tial is high [3,4]. We attempt here to resolve this
where c0 denotes the concentration at x = 0. Fig.
apparent contradiction. The problem is tackled
11 illustrates the variation in c. One can observe
with regard to the relationship between ion accu-
from eq. B2 that the distribution of ions is greatly
mulation and electric field. We shall see that the
affected by the relative magnitudes of J and E,
ion distribution brought about by the ionic flow
since the second term changes sign.
exhibits two kinds of distinct tendencies according
When J is greater than uEc,,, the highest value
to the coupling with electric potential.
of the concentration occurs near x = 0 (case (i) in
The stationary behavior of monovalent cations
fig. 11). Ions accumulate at the efflux site when
is investigated in a one-dimensional system com-
the electric field is weak. The direction of the
prising an aqueous solution with 0 < x =zL, where
diffusion flux given by the first term of eq. Bl is
ion transport occurs at the two boundaries x = 0,
the same as that of the electrically driven flux;
x = L at the rate of flux J. We assume that a
hence, the total flux j (=J) results from the
homogeneous electric field exists along the x-axis;
positive contribution by both fluxes.
this implies that the electric potential V changes
When J is less than uEc,, on the other hand,
linearly with distance (fig. 11). The directions of
ions accumulate near x = L, i.e., near the boundary
flux J and electric field E are assumed to be the
where influx occurs. This is case (ii) in fig. 11. An
same.
intuitive explanation of this result can be made
The concentration of the cation species con-
using eq: Bl: the diffusion flux (in the direction of
cerned is designated c. The flux j is given by
negative x) is in balance with the electrically
j = - D at/ax + UEC, ON driven flux (in the direction of positive x) under
where D represents the diffusion constant of the the condition where j (= J) is small. In the limit-
ion, u electric mobility, and E electric field. The ing case, j = J = 0, this tendency is most clearly
first term arises from diffusion, the latter being evident. Thus, the condition, J < uEc,,, signifies
the electrically driven flux. Concerning the signs that the situation is close to equilibrium. This
of j and E, the x-direction is appointed as posi- phenomenon is analogous to the HalI effect in a
tive. By using the elementary charge e (> 0), the semiconductor. Here, electrons (or holes) accu-
electric current density is equal to ej. In the sta- mulate at the other end under the influence of a
tionary state, flux j must equal the boundary flux Lorentzian force due to the imposed electric cur-
J. The stationary concentration c becomes rent and magnetic field. At equilibrium, the diffu-
sion flux is in balance with the flux due to the
c = J/t.& + ( c0 - J/uE) exp( uEx/D), 032) Lorentzkn force [%I.
174 K. Toko et cll./Electric current loops in plants

Although the electric field is assumed as con- of approx. lo-l4 mol/cm2 per s with pH - 7.0 in
stant, it must be generated self-consistently the acid region under pH 8.0 buffered. Since the
through coupling with the ion distribution. The electric current near the acid region can be re-
electric potential V decreases with x in this model. garded as being composed mainly of Hf via H’
In the weak-field case (i) of J > t&c,, it is possi- pumping [2,15,29,58], the observed electric current
ble that the decrease in electric potential with x may be compared with uEc,; this leads to J = 100
might be due to the flow of ions concerned in this pmol/cm2 per s z+ z&c,. This situation belongs in
situation. The electric current carried by the ions the classification of weak electric field. This sig-
is necessarily accompanied by a potential drop. In nifies that protons accumulate near the efflux site,
the strong-field case (ii) of J < uEc,, it is difficult i.e., in the acid region. Also, in the alkaline region,
for one to suppose that the potential V in fig. 11 similar conclusions may be valid for accumulation
stems from the ions concerned. In fact, the distri- of OH-.
bution of V shows an opposite tendency to that In the case of roots, moderate ion concentra-
expected from the distribution c, which should tion with 1 mM KC1 being present also leads to
evoke a high potential near x = L. The potential V classification of this situation as we+& electric
can be realized only when the electric potential field. This causes the proton accumulation and
due to other origins, which produce the decreasing depletion at the efflux and influx sites, respec-
electric potential with x, overcomes the potential tively, which have been observed using a vibrating
due to the ion distribution c. We can therefore probe 17,351.
suppose that I/ in case (ii) mainly occurs as a
consequence of the ion species differing from those
involved in the present situation, provided the Appendix C
electric field is not intentionally imposed on the
system from the external circuit. Using Einstein’s relation and the stationary
The result is summarized as follows. Even if the conditions ji = Ji and j, = Jk, eq. 6 can be rewrit-
directions of flux and electric field are assumed to ten as
be the same, the concentration distribution un-
dergoes variations according to the relative magni- J;/ui = - ,& &/ax + EC, fori=1,2 ,..., m,
tudes of the flux and field. If the effect of the flux Jk/uk= -pac,/iJx-EC, fork=1,2,...,n,
is greater than that of the electric field, then (Cl)
cations are accumulated at the efflux site; Under
the opposite condition, cations accumulate at the with the definition of p:
part involved in influx. Here, it is possible that the
fi = k,T’/e, (W
electric field or electric potential is mostly due to a
distribution of other ions. This complicated situa- where k, is Boltzmann’s constant and T’ the
tion arises from the fact that the electric current is absolute temperature.
transported by processes expressed as two terms, Electric neutrality is expressed by
viz., diffusion flux and electrically driven flux, as
pointed out earlier [56]. ~c,=~~*c,=c(x,. (C3)
We now comment upon proton accumulation i-l

in the acid region for Chara. The origin of the

The quantity c(x) represents the total ion con-
x-axis is chosen as the midpoint of the acid region,
centration at x. Summation of JJu, in eq. Cl
in which the surface electric potential has a high
leads to
value, The electric current is approx. 10 PA/cm’
[57] and the electric field strength along the cell is
of the order of 25 mV/cm for a change of several C= &,ui= -j3&z,ax+Ec.
i=l
millivolts occurring over a few millimeters along
the cell [3,41]. Calculation of uEc, yields a value The quantity C denotes the total boundary flux of
 K. Toko et ol./ Eiectric current loops in plants 175

cations divided by the electric mobility. This is 9 K. Toko, K. Hayashi and K. Yamafuji, Trans. IECE Jap.
E69 (1986) 485.
independent of x. Similarly, we obtain
10 F.M. Harold, Curr. Top. Membranes Transp. 16 (1982)
n 485.
A= - c Jk/uk=/3 lk/ax+Ec, cab) 11 R.J.P. Williams, J. Theor. Biol. 121 (1986) 1.
k=l 12 R. Larter and P. Ortoleva, J. Theor. Biol. 96 (1982) 175.
13 K. Toko and K. Yamafuji, J. Phys. Sot. Jap. 51 (1982)
where A is a constant relating to the total boundary
3049.
flux of anions. Its sign becomes positive when the 14 K. Toko, T. Fujiyoshi, K. Ggata, H. Chosa and K. Yamafuji,
total flow of anions is net influx at x = 0 (or Biophys. Chem. 27 (1987) 149.
efflux at x = L), which corresponds to an electric 15 K. Toko, H. Chosa and K. Yamafuji, J. Theor. Biol. 114
current flowing from x = 0 to x = L. (1985) 127.
16 P. Gransdorff and I. Prigogine, Thermcdynamic theory of
Combining eqs. C4a and C4b, we obtain structure, stability and fluctuations (Wiley, London, 1971).
T=C+A=2Ec. 17 D.L. Rayle and R.E. Cleland, Plant Physiol. 46 (1970) 250.
(CW
18 J.-M. Verse1 and G. Mayer, Planta 164 (1985) 96.
Eq. CSa implies that the electric field E(x) is of 19 K. Toko, S. Iiyama, C. Tanaka, K. Hayashi, Ke. Yamafuji
positive sign if the quantity T related to the net and Ka. Yamafuji, Biophys. Chem. 27 (1987) 39.
20 T. Yoshida, K. Hayashi, K. Toko and K. Yamafuji, AM.
electric current is positive. This leads to the poten-
Bot. 62 (1988) 497.
tial drop with x. This result is quite reasonable. 21 H. Okamoto, K. Katou and K. Ichino, Plant Cell Physiol.
Subtracting eq. C4a from eq. C4b, we obtain 20 (1979) 103.
22 J.B. Hanson, Plant Physiol. 62 (1978) 402.
S= C-A= -2fi &/ax, (C5b) 23 A.H. de Boer, H.B.A. Prins and P.E. Zanstra, Planta 157
(1983) 259.
which signifies that the ion concentration near
24 AS. Crafts and T.C. Brayer, Am. J. Bot. 25 (1938) 529.
x = 0 is high if the total ion flux occurs in the 25 W.H. Arisz, Protoplasma 46 (1956) 5.
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Eq. C5b can be easily integrated, since S is a 28 K. Ogata, Plant Cell Physiol. 24 (1983) 695.
29 K. Ogata, K. Toko, T. Fujiyoshi and K. Yatnafuji, Biophys.
constant composed of the boundary fluxes, and
Chem. 26 (1987) 71.
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C5a, we derive E(x). The concentration of each 14 (1973) 127.
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151.
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